Environmental factors and chemical and microbiological water-quality constituents related to the presence of enteric viruses in ground water from small public water supplies in southeastern Michigan

USGS Publications Warehouse

Francy, Donna S.; Bushon, Rebecca N.; Stopar, Julie; Luzano, Emma J.; Fout, G. Shay

2004-01-01

A study of small public ground-water-supply wells that produce water from discontinuous sand and gravel aquifers was done from July 1999 through July 2001 in southeastern Michigan. Samples were collected to determine the occurrence of viral pathogens and microbiological indicators of fecal contamination (indicators), determine whether indicators are adequate predictors of the presence of enteric viruses, and determine the factors that affect the presence of enteric viruses. Small systems are those that serve less than 3,300 people. Samples were analyzed for specific enteric viruses by reverse transcriptase-polymerase chain reaction (RT-PCR), for culturable viruses by cell culture, and for the indicators total coliforms, Escherichia coli (E. coli), enterococci, and F-specific and somatic coliphage. Ancillary environmental and water-quality data were collected or compiled. A total of 169 regular samples and 32 replicate pairs were collected from 38 wells. Replicate pairs were samples collected at the same well on the same date. One well was sampled 6 times, 30 wells were sampled five times, 6 wells were sampled twice, and 1 well was sampled once. By use of RT-PCR, enterovirus was found in four wells (10.5 percent) and hepatitis A virus (HAV) in five wells (13.2 percent). In two of these wells, investigators found both enterovirus and HAV, but on different sampling dates. Culturable viruses were found one time in two wells (5.9 percent), and neither of these wells was positive for viruses by use of RT-PCR on any sampling date. If results for all viruses are combined, 9 of the 38 small public-supply wells were positive for enteric viruses (23.7 percent) by either cell culture or RT-PCR. One or more indicators were found in 18 of 38 wells. Total coliforms, E. coli, enterococci, and F-specific and somatic coliphage were found in 34.2, 10.5, 15.8, 5.9, and 5.9 percent, respectively, of the wells tested. In only 3 out of 18 wells were samples positive for an indicator on more than one date at the same well. The co-occurrence of enteric viruses and any indicator was 55.6 percent; five out of the nine virus-positive wells were also found to be positive for an indicator. Two wells with detections of viruses had a detection of total coliforms, one well had a detection of E. coli, one of enterococci, and one of F-specific coliphage. On a per sample basis, of 11 samples that were positive for enteric viruses, indicator bacteria co-occurred in only 2 samples, and coliphage were not present in any. More virus-positive samples were found at sites served by septic systems than those served by sewerlines. Sampling condition (ground water or a mixture of tank and ground water), distance to septic system, type of and distance to nearest surface-water body, well characteristics, or land use were not related to the presence of viruses or indicators. Among continuous water-quality variables, statistically significant relations were found between total coliforms and dissolved organic carbon and between total coliforms and iron. There was a statistically significant relation between chloride concentrations >20 mg/L and detections of total coliforms. Presence of nitrate and nitrite was related to the presence of other indicators (E. coli, enterococci, and F-specific and somatic coliphage) or enteric viruses, but not to total coliforms. The data indicated that chloride-to-bromide (C1:Br) ratios may be useful as a screening tool for total coliforms and enteric viruses but not for E. coli, enterococci, and F-specific and somatic coliphage. This study provides evidence for fecal contamination of ground water from small public-supply wells, at least on an intermittent basis. Collecting data on multiple lines of evidence would be needed to reliably predict the presence of enteric viruses and protect public health. Future data collection toward this end could include repeat sampling several times a year for different indicators, measuring dissolv

Detection of Cryptosporidium and Cyclospora Oocysts from Environmental Water for Drinking and Recreational Activities in Sarawak, Malaysia.

PubMed

Bilung, Lesley Maurice; Tahar, Ahmad Syatir; Yunos, Nur Emyliana; Apun, Kasing; Lim, Yvonne Ai-Lian; Nillian, Elexson; Hashim, Hashimatul Fatma

2017-01-01

Cryptosporidiosis and cyclosporiasis are caused by waterborne coccidian protozoan parasites of the genera Cryptosporidium and Cyclospora, respectively. This study was conducted to detect Cryptosporidium and Cyclospora oocysts from environmental water abstracted by drinking water treatment plants and recreational activities in Sarawak, Malaysia. Water samples (12 each) were collected from Sungai Sarawak Kanan in Bau and Sungai Sarawak Kiri in Batu Kitang, respectively. In addition, 6 water samples each were collected from Ranchan Recreational Park and UNIMAS Lake at Universiti Malaysia Sarawak, Kota Samarahan, respectively. Water physicochemical parameters were also recorded. All samples were concentrated by the iron sulfate flocculation method followed by the sucrose floatation technique. Cryptosporidium and Cyclospora were detected by modified Ziehl-Neelsen technique. Correlation of the parasites distribution with water physicochemical parameters was analysed using bivariate Pearson correlation. Based on the 24 total samples of environmental water abstracted by drinking water treatment plants, all the samples (24/24; 100%) were positive with Cryptosporidium , and only 2 samples (2/24; 8.33%) were positive with Cyclospora . Based on the 12 total samples of water for recreational activities, 4 samples (4/12; 33%) were positive with Cryptosporidium , while 2 samples (2/12; 17%) were positive with Cyclospora . Cryptosporidium oocysts were negatively correlated with dissolved oxygen (DO).

Detection of Cryptosporidium and Cyclospora Oocysts from Environmental Water for Drinking and Recreational Activities in Sarawak, Malaysia

PubMed Central

Tahar, Ahmad Syatir; Yunos, Nur Emyliana; Apun, Kasing; Nillian, Elexson; Hashim, Hashimatul Fatma

2017-01-01

Cryptosporidiosis and cyclosporiasis are caused by waterborne coccidian protozoan parasites of the genera Cryptosporidium and Cyclospora, respectively. This study was conducted to detect Cryptosporidium and Cyclospora oocysts from environmental water abstracted by drinking water treatment plants and recreational activities in Sarawak, Malaysia. Water samples (12 each) were collected from Sungai Sarawak Kanan in Bau and Sungai Sarawak Kiri in Batu Kitang, respectively. In addition, 6 water samples each were collected from Ranchan Recreational Park and UNIMAS Lake at Universiti Malaysia Sarawak, Kota Samarahan, respectively. Water physicochemical parameters were also recorded. All samples were concentrated by the iron sulfate flocculation method followed by the sucrose floatation technique. Cryptosporidium and Cyclospora were detected by modified Ziehl-Neelsen technique. Correlation of the parasites distribution with water physicochemical parameters was analysed using bivariate Pearson correlation. Based on the 24 total samples of environmental water abstracted by drinking water treatment plants, all the samples (24/24; 100%) were positive with Cryptosporidium, and only 2 samples (2/24; 8.33%) were positive with Cyclospora. Based on the 12 total samples of water for recreational activities, 4 samples (4/12; 33%) were positive with Cryptosporidium, while 2 samples (2/12; 17%) were positive with Cyclospora. Cryptosporidium oocysts were negatively correlated with dissolved oxygen (DO). PMID:29234679

HIV-positive MSM's knowledge of HPV and anal cancer self-sampling: A scoping review.

PubMed

Poon, M K L; Wong, J P H; Li, A T W; Manuba, M; Bisignano, A; Owino, M; Vahabi, M

2018-02-01

Human papillomavirus (hpv) infection is the cause of anal squamous cell cancer (ascc) in 80% of cases. Available research has also shown high prevalence of anal hpv infection among men who have sex with men (msm). However, hpv vaccination is low among msm in Canada. In light of this information, we conducted a scoping review with the aim of exploring (1) the knowledge of hpv and anal cancer among hiv-positive msm and (2) the acceptability of hpv and anal cancer self-sampling in this population. In conducting the review, we searched five electronic databases for peer-reviewed articles and abstracts published in English, between 2007 and 2017. A total of 803 articles were retrieved; after accounting for duplicates ( n= 40) and unmet criteria ( n= 754), a total of 794 articles were excluded. A final total of nine articles were used in this review. Results of this review show that hiv-positive msm have limited knowledge regarding the risks of anal cancer associated with hiv and hpv coinfection. Furthermore, there is limited research on hpv and anal cancer self-sampling in this population. However, the review of available studies suggested that hiv-positive msm were open to anal cancer self-sampling. It also identified potential barriers to self-sampling. In conclusion, we provide suggestions and future directions for policy-makers and educators to develop inclusive and accessible strategies to reach hiv-positive msm regarding anal cancer education and self-screening.

HIV-positive MSM’s knowledge of HPV and anal cancer self-sampling: A scoping review

PubMed Central

Poon, M.K.L.; Wong, J.P.H.; Li, A.T.W.; Manuba, M.; Bisignano, A.; Owino, M.; Vahabi, M.

2018-01-01

Human papillomavirus (hpv) infection is the cause of anal squamous cell cancer (ascc) in 80% of cases. Available research has also shown high prevalence of anal hpv infection among men who have sex with men (msm). However, hpv vaccination is low among msm in Canada. In light of this information, we conducted a scoping review with the aim of exploring (1) the knowledge of hpv and anal cancer among hiv-positive msm and (2) the acceptability of hpv and anal cancer self-sampling in this population. In conducting the review, we searched five electronic databases for peer-reviewed articles and abstracts published in English, between 2007 and 2017. A total of 803 articles were retrieved; after accounting for duplicates (n=40) and unmet criteria (n=754), a total of 794 articles were excluded. A final total of nine articles were used in this review. Results of this review show that hiv-positive msm have limited knowledge regarding the risks of anal cancer associated with hiv and hpv coinfection. Furthermore, there is limited research on hpv and anal cancer self-sampling in this population. However, the review of available studies suggested that hiv-positive msm were open to anal cancer self-sampling. It also identified potential barriers to self-sampling. In conclusion, we provide suggestions and future directions for policy-makers and educators to develop inclusive and accessible strategies to reach hiv-positive msm regarding anal cancer education and self-screening. PMID:29507499

Accuracy of BRT and Delvotest microbial inhibition tests as affected by composition of ewe's milk.

PubMed

Althaus, Rafael; Torres, Antonio; Peris, Cristofol; Beltran, M Carmen; Fernandez, Nemesio; Molina, M Pilar

2003-03-01

The presence of drug residues in ewe's milk samples can be determined by microbial assays. The main limitation of these tests is the large number of false-positive results associated with them. False-positive results can be explained by the interaction of certain substances naturally existing in ewe's milk with the growth of the microorganism used in the test. In this study, milk chemical composition (fat, protein, lactose, total solids), somatic cell counts (SCCs), free fatty acid concentrations, and lactoperoxidase system components were determined in order to investigate their influence on the rate of false-positive results for the BRT and Delvotest microbiological inhibitor tests. Milk samples were obtained after morning milking of Manchega ewes at 15, 30, 45, 60, 75, 90, 105, 120, and 135 days after parturition. The animals did not receive any kind of treatment or medicated feed throughout the experiment. The false-positive rates for BRT and Delvotest were 3.75 and 2.4%, respectively. When the logistic regression model was applied, the percentages of total solids for positive samples were significantly different from those for negative samples (16.90 versus 18.42% for BRT, 16.05 versus 18.45% for Delvotest), while the SCC logarithmic transformation was significantly higher for the positive samples than for the negative samples (5.38 versus 5.11 log units for BRT, 5.32 versus 5.11 log units for Delvotest). Moreover, Delvotest-positive samples exhibited thiocyanate concentrations higher than those of Delvotest-negative samples (8.18 mg/liter versus 6.85 mg/liter). Further analyses are needed to confirm the possible presence of antimicrobial residues in this particular type of milk sample.

Detection of classical swine fever virus E2 gene in cattle serum samples from cattle herds of Meghalaya.

PubMed

Chakraborty, A K; Karam, A; Mukherjee, P; Barkalita, L; Borah, P; Das, S; Sanjukta, R; Puro, K; Ghatak, S; Shakuntala, I; Sharma, I; Laha, R G; Sen, A

2018-03-01

The present study focused on the detection and genetic characterisation of 5' untranslated region (5'UTR) and E2 gene of classical swine fever virus (CSFV, family Flaviviridae , genus Pestivirus ) from bovine population of the northeastern region of India. A total of 134 cattle serum samples were collected from organised cattle farms and were screened for CSFV antigen with a commercial antigen capture enzyme linked immunosorbent assay (Ag-ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). A total of 10 samples were positive for CSFV antigen by ELISA, while all of them were positive in PCR for 5'UTR region. Full length E2 region of CSFV were successfully amplified from two positive samples and used for subsequent phylogenetic analysis and determination of protein 3D structure which showed similarity with reported CSFV isolate from Assam of sub-genogroup 2.1, with minor variations in protein structure.

Microbiological baseline study of beef and pork carcasses from provincially inspected abattoirs in Alberta, Canada

PubMed Central

Bohaychuk, Valerie M.; Gensler, Gary E.; Barrios, Pablo Romero

2011-01-01

In 2006 and 2007 beef and pork carcass swabs from provincially inspected abattoirs in Alberta, Canada were tested to determine the levels of total aerobic bacteria, coliform bacteria, and generic Escherichia coli, and the prevalence of Salmonella spp., Campylobacter spp., and Shiga toxin-producing E. coli (STEC). Swabs from beef and pork carcasses from 48 and 34 facilities, respectively, were analyzed. All samples tested were positive for aerobic bacteria with 99.8% of beef and 96.0% of pork samples, having total counts of ≤ 100 000 CFU/cm2. Coliform bacteria were isolated from 22.4% and 42.0% of beef and pork carcass samples, respectively. Generic E. coli were recovered from 14.6% of beef and 33.7% of pork carcass samples. For beef carcasses, positive tests were obtained for 0.1% of 1036 samples tested for Salmonella spp., 1.5% of 1022 samples tested for Campylobacter spp. and 5.4% of 1018 samples tested for STEC. For pork carcasses, positive tests were obtained for 1.6 % of 1076 samples tested for Salmonella spp., 8.8% of 1070 samples tested for Campylobacter spp. and 4.8% of 1067 samples tested for STEC. PMID:22467964

Epidemiology of hepatitis E virus infection during pregnancy in Benin.

PubMed

De Paschale, Massimo; Ceriani, Cristina; Romanò, Luisa; Cerulli, Teresa; Cagnin, Debora; Cavallari, Serena; Ndayake, Joseph; Zaongo, Dieudonné; Diombo, Kouma; Priuli, Gianbattista; Viganò, Paolo; Clerici, Pierangelo

2016-01-01

Hepatitis E virus (HEV) is the cause of enterically transmitted non-A, non-C hepatitis (an infection that is particularly severe during pregnancy) in tropical and subtropical countries. As there are no published data concerning the prevalence of HEV antibodies in Benin, their presence was investigated in pregnant women undergoing routine HIV screening in a rural area in northern Benin and in pregnant women with acute non-A, non-C hepatitis. A total of 278 serum samples were collected from asymptomatic pregnant women in 2011 were tested for HEV and hepatitis A virus (HAV) antibodies, and the HEV IgM-positive samples were further tested for HEV-RNA. A further seven samples of pregnant women with acute non-A, non-C hepatitis collected during episodes of acute hepatitis in 2005 were also analysed. Of the 278 samples collected in 2011, 16.19% were positive for HEV IgG and 1.44% for HEV IgM (none positive for HEV-RNA), and 99.64% were positive for total HAV antibodies (none positive for HAV IgM). Six of the seven samples collected in 2005 were positive for HEV IgG and IgM, and two were also positive for HEV-RNA. The circulation of HEV infection is significant among pregnant women in Benin, in whom the consequences may be fatal. © 2015 John Wiley & Sons Ltd.

17 CFR Appendix B to Part 420 - Sample Large Position Report

Code of Federal Regulations, 2010 CFR

2010-04-01

..., and as collateral for financial derivatives and other securities transactions $ Total Memorandum 1... 17 Commodity and Securities Exchanges 3 2010-04-01 2010-04-01 false Sample Large Position Report B Appendix B to Part 420 Commodity and Securities Exchanges DEPARTMENT OF THE TREASURY REGULATIONS UNDER...

Investigation of zoonotic disease pathogens (Aeromonas hydrophila, Pseudomonas fluorescens, Streptococcus iniae) seen in carp farms in the Northern Iraq-Erbil region by molecular methods

NASA Astrophysics Data System (ADS)

Ibraheem, Azad Saber; Önalan, Şükrü; Arabacı, Muhammed

2017-04-01

The aim of this study was to determine the zoonotic bacteria in carp farms in the Northern Iraq-Erbil region. Carp is the main fish species cultured in Erbil region. The most common zoonotic bacteria generally seen in carp farms are Aeromonas hydrophila, Pseudomonas fluorescens and Streptococcus iniae. Samples were collected from 25 carp farms in the Northern Iraq-Erbil region. Six carp samples were collected from each carp farm. Head kidney and intestine tissue samples were collected from each carp sample. Then head kidney and intestine tissue samples were pooled separately from each carp farm. Total bacterial DNA had been extracted from the 25 pooled head kidney and 25 intestinal tissue samples. The pathogen Primers were originally designed from 16S RNA gene region. Zoonotic bacteria were scanned in all tissue samples with absent/present analysis by RT-PCR. Furthermore, the capillary gel electrophoresis bands were used for confirmation of amplicon size which was planned during primer designing stage. As a result, thirteen carp farms were positive in the respect to Aeromonas hydrophila, eight carp farms were positive from head kidney and six carp farms were positive from the intestine, only one carp farm was positive from both head kidney and the intestine tissue samples. In the respect to Streptococcus iniae, four carp farms were positive from head kidney and two carp farms were positive from the intestine. Only one carp farm was positive in the respect to Pseudomonas fluorescens from the intestine. Totally, 9 of 25 carp farms were cleared (negative) the zoonotic bacteria. In conclusion, the zoonotic bacteria were high (64 %) in carp farms in the Northern Iraq-Erbil region.

40 CFR 142.16 - Special primacy requirements.

Code of Federal Regulations, 2011 CFR

2011-07-01

... coliform or E. coli testing on a total coliform-positive sample)—A description of how the State will determine whether it is appropriate to waive fecal coliform or E. coli testing on a total coliform-positive... where allowed in State programs. (1) Approve an alternative to the E. coli levels that trigger...

Microbiological quality of fresh produce obtained from retail stores on the Eastern Shore of Maryland, United States of America.

PubMed

Korir, Robert Cheruiyot; Parveen, Salina; Hashem, Fawzy; Bowers, John

2016-06-01

The aim of this study was to investigate the microbiological quality of six types of fresh produce obtained from three retail stores located on the Eastern Shore of Maryland, USA. A total of 414 samples representing basil, cilantro, lettuce, scallion, spinach, and parsley were analyzed for total aerobic bacteria (APC), total coliforms, Escherichia coli, and three pathogenic bacteria (E. coli O157:H7, Listeria monocytogenes, and Salmonella), using standard methods. Presumptive pathogenic isolates were confirmed using BAX Polymerase Chain Reaction. Total aerobic populations varied widely between samples, while 38.41% were positive for total coliforms and only 10.15% for E. coli. Median abundance (log CFU/g) of total coliforms and E. coli were less than the limit of detection and that of APC ranged from 5.78 to 6.61 over the six produce types. There was a statistically significant difference in prevalence of total coliforms among the retail stores, but not for abundance of APC or prevalence of E. coli. E. coli O157:H7 and L. monocytogenes were detected in one spinach sample each, while one parsley and one cilantro sample were positive for Salmonella. There were no statistically significant differences in microbiological quality among produce types. Although the results of this study provided some indices of sanitary and/or spoilage level, no relationship was observed among the total aerobic bacteria, total coliforms, E. coli, and the presence of pathogenic bacteria in the samples tested. Copyright © 2015 Elsevier Ltd. All rights reserved.

Characterizing the temporal patterns of avian influenza virus introduction into Japan by migratory birds

PubMed Central

ONUMA, Manabu; KAKOGAWA, Masayoshi; YANAGISAWA, Masae; HAGA, Atsushi; OKANO, Tomomi; NEAGARI, Yasuko; OKANO, Tsukasa; GOKA, Koichi; ASAKAWA, Mitsuhiko

2017-01-01

The objectives of the present study were to observe the temporal pattern of avian influenza virus (AIV) introduction into Japan and to determine which migratory birds play an important role in introducing AIV. In total, 19,407 fecal samples from migratory birds were collected at 52 sites between October 2008 and May 2015. Total nucleic acids extracted from the fecal samples were subjected to reverse transcription loop–mediated isothermal amplification to detect viral RNA. Species identification of host migratory birds was conducted by DNA barcoding for positive fecal samples. The total number of positive samples was 352 (prevalence, 1.8%). The highest prevalence was observed in autumn migration, and a decrease in prevalence was observed. During autumn migration, central to southern Japan showed a prevalence higher than the overall prevalence. Thus, the main AIV entry routes may involve crossing the Sea of Japan and entry through the Korean Peninsula. Species identification was successful in 221 of the 352 positive samples. Two major species sequences were identified: the Mallard/Eastern Spot-billed duck group (115 samples; 52.0%) and the Northern pintail (61 samples; 27.6%). To gain a better understanding of the ecology of AIV in Japan and the introduction pattern of highly pathogenic avian influenza viruses, information regarding AIV prevalence by species, the prevalence of hatch-year migratory birds, migration patterns and viral subtypes in fecal samples using egg inoculation and molecular-based methods in combination is required. PMID:28484128

Effect of Type of Enrichment and Duration of Incubation on Salmonella Recovery from Meat-and-Bone Meal

PubMed Central

Huhtanen, C. N.; Naghski, J.

1972-01-01

Twenty-five meat-and-bone meal samples were enriched with either selenite-cystine or tetrathionate and incubated for 1 and 2 days. Seven were previously found to be positive; of the other 18, 16 were positive for salmonella. The number of somatic serogroups per sample ranged from 1 to 11 with a mean of 3.8. Significantly more (P < 0.01) group C1 salmonellae were isolated using tetrathionate than selenite, whereas significantly more of groups G, 35, and Difco poly-valent D were isolated from selenite than tetrathionate. Seventy-six percent of the presumptive colonies from Brilliant Green agar showed a positive lysine decarboxylase reaction, and there were no differences between media or times of incubation. Ninety-four per cent of the lysine decarboxylase-positive cultures showed a positive somatic antiserum response; again there were no differences between times or enrichments although there were significantly more total positive serogroups at 2 days than at 1 day from tetrathionate but not from selenite. There were indications that certain serogroups preferred either one or the other enrichment. There were no differences in total positive samples with the two enrichments although neither alone was sufficient to identify all positives. Several lactose-positive salmonellae were recovered. PMID:4553803

Application of the BAX for screening/genus Listeria polymerase chain reaction system for monitoring Listeria species in cold-smoked fish and in the smoked fish processing environment.

PubMed

Norton, D M; McCamey, M; Boor, K J; Wiedmann, M

2000-03-01

The cold-smoked fish industry was used as a model for the development of a system for monitoring Listeria spp. in foods and in the food processing environment. A total of 214 samples including raw fish, fish during the cold-smoking process, finished product, and environmental samples were collected from three processing facilities over two visits to each facility. Samples were screened for Listeria spp. using the BAX for Screening/genus Listeria polymerase chain reaction system (PCR) and by culture. Listeria spp., confirmed by the API Listeria test strip or by a PCR assay targeting the L. monocytogenes hlyA gene, were isolated from a total of 89 (41.6%) samples. Of these, 80 samples also tested positive for Listeria spp. using the BAX system. Specifically, 42 (55.3%) environmental samples (n = 76), 11 (25.6%) raw materials samples (n = 43), 20 (35.1%) samples from fish in various stages of processing (n = 57), and 7 (18.4%) finished product samples (n = 38) tested positive for Listeria spp. using the BAX system. Five (4.0%) of the 125 culture-negative samples yielded BAX system-positive results. Listeria isolates from each of nine culture-positive/BAX system-negative samples yielded a positive reaction when tested in pure culture by the BAX system, suggesting that our false-negative results were likely due to the presence of low Listeria numbers in the initial enrichment as opposed to nonreacting isolates. The employment of alternative enrichment protocols, such as the two-step enrichment recommended by the manufacturer, may increase the sensitivity of the assay.

Microbial Contamination of Human Milk Purchased Via the Internet

PubMed Central

Hogan, Joseph S.; McNamara, Kelly A.; Gudimetla, Vishnu; Dillon, Chelsea E.; Kwiek, Jesse J.; Geraghty, Sheela R.

2013-01-01

OBJECTIVE: To quantify microbial contamination of human milk purchased via the Internet as an indicator of disease risk to recipient infants. METHODS: Cross-sectional sample of human milk purchased via a popular US milk-sharing Web site (2012). Individuals advertising milk were contacted to arrange purchase, and milk was shipped to a rented mailbox in Ohio. The Internet milk samples (n = 101) were compared with unpasteurized samples of milk donated to a milk bank (n = 20). RESULTS: Most (74%) Internet milk samples were colonized with Gram-negative bacteria or had >104 colony-forming units/mL total aerobic count. They exhibited higher mean total aerobic, total Gram-negative, coliform, and Staphylococcus sp counts than milk bank samples. Growth of most species was positively associated with days in transit (total aerobic count [log10 colony-forming units/mL] β = 0.71 [95% confidence interval: 0.38–1.05]), and negatively associated with number of months since the milk was expressed (β = −0.36 [95% confidence interval: −0.55 to −0.16]), per simple linear regression. No samples were HIV type 1 RNA-positive; 21% of Internet samples were cytomegalovirus DNA-positive. CONCLUSIONS: Human milk purchased via the Internet exhibited high overall bacterial growth and frequent contamination with pathogenic bacteria, reflecting poor collection, storage, or shipping practices. Infants consuming this milk are at risk for negative outcomes, particularly if born preterm or are medically compromised. Increased use of lactation support services may begin to address the milk supply gap for women who want to feed their child human milk but cannot meet his or her needs. PMID:24144714

Detection of Toxoplasma gondii in the milk of naturally infected goats in the Northeast of Brazil.

PubMed

Bezerra, M J G; Kim, P C P; Moraes, É P B X; Sá, S G; Albuquerque, P P F; Silva, J G; Alves, B H L S; Mota, R A

2015-08-01

The aim of the present study was to detect the genomic DNA of Toxoplasma gondii in milk samples from naturally infected goats in the state of Pernambuco, (Brazil). In total, 248 blood serum samples were collected and processed from lactating goats and then submitted to a search for antibodies to T. gondii through the indirect immunofluorescence reaction. Samples with a score of 64 or more were considered positive. In total, 248 milk samples were collected and processed from the same group of goats in order to study the DNA of T. gondii using the polymerase chain reaction (PCR) technique. In the serum samples, 56/248 (22.58%) of the animals were positive, whereas the DNA of the parasite was detected in 15/248 (6.05%) of the milk samples. Five of these 15 samples were animals who were also positive in the serology. This study reports the first occurrence of the elimination of T. gondii from the milk of naturally infected goats in the north-east of Brazil. It is suggested that the consumption of in natura goat milk may constitute a potential risk to the health of milk consumers in this region. © 2013 Blackwell Verlag GmbH.

Investigation for zoonotic disease pathogens (Aeromonas hydrophila, Pseudomonas fluorescens, Streptococcus iniae) seen in carp farms in Duhok region of Northern Iraq by molecular methods

NASA Astrophysics Data System (ADS)

Mohammed, Kamiran Abdulrahman; Arabacı, Muhammed; Önalan, Şükrü

2017-04-01

The aim of this study was to determine the zoonotic bacteria in carp farms in Duhok region of the Northern Iraq. Carp is the main fish species cultured in the Duhok region. The most common zoonotic bacteria generally seen in carp farms are Aeromonas hydrophila, Pseudomonas fluorescens and Streptococcus iniae. Samples were collected from 20 carp farms in the Duhok Region of the Northern Iraq. Six carp samples were collected from each carp farm. Head kidney tissue samples and intestine tissue samples were collected from each carp sample. Than head kidney and intestine tissue samples were pooled. The total bacterial DNA extraction from the pooled each 20 head kidney tissue samples and pooled each 20 intestinal tissue samples. Primers for pathogens were originally designed from 16S Ribosomal gene region. Zoonotic bacteria were scanned in all tissue samples by absent / present analysis in the RT-PCR. After RT-PCR, Capillary gel electrophoresis bands were used for the confirmation of the size of amplicon which was planned during primer designing stage. As a result, one sample was positive in respect to Aeromonas hydrophila, from intestine and one carp farm was positive in respect to Pseudomonas fluorescens from intestine and two carp farms were positive in respect to Streptococcus iniae. Totally 17 of 20 carp farms were negative in respect to the zoonotic bacteria. In conclusion the zoonotic bacteria were very low (15 %) in carp farms from the Duhok Region in the Northern Iraq. Only in one Carp farms, both Aeromonas hydrophila and Pseudomonas fluorescens were positive. Also Streptococcus inia were positive in two carp farms.

Children’s Caregivers and Public Playgrounds: Potential Reservoirs of Infection of Hand-foot-and-mouth Disease

NASA Astrophysics Data System (ADS)

Li, Pengyuan; Li, Tao; Gu, Qiuyun; Chen, Xiaomin; Li, Jiahui; Chen, Xiashi; Chen, Yan; Zhang, Danwei; Gao, Rong; He, Zhenjian; Zhu, Xun; Zhang, Wangjian; Hao, Yuantao; Zhang, Dingmei

2016-11-01

Hand-foot-and-mouth disease (HFMD) is a common infectious disease, which has led to millions of clinical cases and hundreds of deaths every year in China. This study aimed to exploring the effects on HFMD transmission of children’s caregivers and public area, as well as trying to locate the potential reservoirs of infections in primary cases. Total children’s 257 samples (98 children’s caregivers and 159 environmental samples) were tested for the presence of universal enterovirus, enterovirus 71, coxsackie virus A6 and A16 by real-time fluorescence quantitative polymerase chain reaction (qPCR). 5.84% (15/257, 95% confidence interval [CI]: 2.98%, 8.70%) of total samples had positive results of enterovirus. The enterovirus positive rates of children’s caregiver samples and environmental samples were respectively 7.14% (7/98, 95% CI: 2.04%, 12.24%), and 5.03% (8/159, 95% CI: 1.63%, 8.43%); 7.61% (7/92, 95% CI: 2.21%, 13.01%) of wiping samples from playgrounds and 1.49% (1/67, 95% CI: 0, 7.00%) of air samples in indoor market places had positive result of enterovirus. High positive rates of enterovirus in children’s caregivers and from playgrounds indicated that they would be potential reservoirs of HFMD infection, as children might be infected via contacting with asymptomatic-infected individuals or exposure of contaminated surface of public facilities.

A molecular survey of bovine Theileria parasites among apparently healthy cattle and with a note on the distribution of ticks in eastern Turkey.

PubMed

Aktas, Munir; Altay, Kursat; Dumanli, Nazir

2006-06-15

A survey of Theileria parasites in cattle in eastern Turkey was carried out using specific polymerase chain reaction. A total of 252 blood samples were collected from clinically healthy cattle between June and July 2004. Of 252 blood samples examined, 41 (16%) were positive for piroplasms by microscopy, whereas 114 (45%) were positive for the presence of at least one species of Theileria by PCR. The percentages of positive animals for Theileria annulata and benign Theileria species (Theileria sergenti/buffeli/orientalis) were 39% (99/252) and 7% (18/252), respectively. By allele-specific PCR examination of 18 field isolates which were positive for benign Theileria parasites, 8 samples were only amplified by B-type specific primers and 10 samples were amplified by both of the B and C-type specific primers, indicating a mixed infection with B and C-type of the parasite. None of the field isolates was amplified by I-type specific primers. Three samples were co-infected with T. annulata and benign Theileria parasites. Two of them which were infected with B-type parasite were also infected with T. annulata, the other sample which was infected both of B and C-type parasites was also infected with T. annulata. A total of 724 ixodid ticks were collected from the cattle. Hyalomma anatolicum anatolicum was the dominant species with 32% (230/724) in the region. H. a. excavatum, Boophylus annulatus and Rhipicephalus bursa represented 25% (183/724), 19% (140/724) and 15% (112/724) of the total number of ticks, respectively. R. sanguineus was the minor species and represented 8% (59/724) of the tick population.

Excoriation (skin-picking) disorder in adults: a cross-cultural survey of Israeli Jewish and Arab samples.

PubMed

Leibovici, Vera; Koran, Lorrin M; Murad, Sari; Siam, Ihab; Odlaug, Brian L; Mandelkorn, Uri; Feldman-Weisz, Vera; Keuthen, Nancy J

2015-04-01

We sought to estimate the lifetime prevalence of Excoriation (Skin-Picking) Disorder (SPD) in the Israeli adult population as a whole and compare SPD prevalence in the Jewish and Arab communities. We also explored demographic, medical and psychological correlates of SPD diagnosis. Questionnaires and scales screening for SPD, and assessing the severity of perceived stress, depression, obsessive-compulsive disorder (OCD), body dysmorphic disorder (BDD), alcohol use, illicit drug use, and medical disorders were completed in a sample of 2145 adults attending medical settings. The lifetime prevalence of SPD was 5.4% in the total sample; it did not differ between genders or within Jewish and Arab subsamples. Severity of depression (p<0.001), OCD (p<0.001) and perceived stress (p=<0.001) were greater in the SPD positive sample. Similarly, diagnoses of BDD (p=0.02) and generalized anxiety (p=0.03) were significantly more common in the SPD-positive respondents. Alcohol use and illicit substance use were significantly more common among SPD positive respondents in the total sample (both p's=0.01) and the Jewish subsample (p=0.03 and p=0.02, respectively). Hypothyroidism was more prevalent in the SPD-positive Jewish subsample (p=0.02). In the total sample, diabetes mellitus was more common in women than in men (p=0.04). Lifetime SPD appears to be relatively common in Israeli adults and associated with other mental disorders. Differences in the self-reported medical and psychiatric comorbidities between the Jewish and Arab subsamples suggest the possibility of cross-cultural variation in the correlates of this disorder. Copyright © 2014 Elsevier Inc. All rights reserved.

Determination of levels of polychlorinated biphenyl in transformers oil from some selected transformers in parts of the Greater Accra Region of Ghana.

PubMed

Buah-Kwofie, Archibold; Yeboah, Philip O; Pwamang, John

2011-01-01

Although polychlorinated biphenyls have never been manufactured in Ghana, it has been used extensively as dielectric fluid in electric transformers and capacitors. However, very little is known of its health and environmental impacts by both managers of these transformers and capacitors and also the general public. This work therefore seeks to explore INAA as a possible alternative to screening transformer oils for PCBs by determining the total chlorine content. The total chlorine content of transformer oil samples from Ghana that tested positive and some randomly selected samples that tested negative from screening using CLOR-N-OIL test kits, have had their total chlorine estimated. INAA using the Research Reactor located at the Ghana Atomic Energy Commission was used to estimate the total chlorine content of the oil samples. Neutron Activation and gamma ray spectroscopy using HPGe detector coupled to MAESTRO 32 software was used to determine the total chlorine content by integrating the peak area of the spectrum into a simplified program that was developed from the activation equation. Instrumental Neutron Activation Analysis was able to validate the result obtained from the test kits screening with accuracy 7.5%. The minimum total chlorine content of the positive samples determined by NAA was 71.34 μg g⁻¹. Copyright © 2010 Elsevier Ltd. All rights reserved.

Study of Umbilical Cord Blood Culture in Diagnosis of Early-onset Sepsis Among Newborns with High-risk Factors.

PubMed

Kalathia, Mitul Babubhai; Shingala, Prakash Ashokbhai; Parmar, Parin Niranjanbhai; Parikh, Yogesh Narenedrabhai; Kalathia, Ila Mitulkumar

2013-10-01

Blood culture is gold standard for diagnosis of neonatal sepsis. Low sensitivity of blood culture is usually due to small volume of blood sample, intrapartum antibiotics, and antibiotics given to newborn before sampling. We evaluated use of Umbilical cord blood culture (UCBC) in diagnosis of neonatal sepsis as compared to peripheral venous blood culture. This study was done in tertiary care teaching hospital during May-June 2012. A total of 45 newborns with presence of two or more risk factors of sepsis were included. Blood sample from placental end of umbilical cord was collected and cultured. Primary outcome was diagnosis of neonatal sepsis by use of umbilical cord blood sample as compared with venous blood sample. Secondary outcome was to compare organisms identified by UCBC and venous blood culture. Sensitivity, specificity, positive and negative predictive values of UCBC were calculated. A total of 24.44% (11 out of 45) high-risk newborns had positive UCBC. A total of 17.8% (8 out of 45) newborns had positive blood culture report. Organisms grown in UCBC were Pseudomonas (45%, 5 out of 11), Acinetobacter (27.27%, 3 out of 11), Escherichia coli (18.18%, 2 out of 11), and Klebsiella (9%, 1 out of 11). UCBC is a good method for diagnosis of neonatal sepsis among high-risk newborns as compared to venous blood culture with a sensitivity of 80% and specificity of 91.43%. Organisms grown are comparable to blood culture samples.

Influenza A virus among the hospitalized young children with acute respiratory infection. Is influenza A co infected with respiratory syncytial virus?

PubMed Central

Alavi, Seyed Mohammad; Makvandi, Manoochehr; Najafi-Fard, Saied; Alavi, Leila

2012-01-01

Background: Both influenza A virus (IAV) and respiratory syncytial virus (RSV) cause acute respiratory infection (ARI) in infants and young children. This study was conducted to determine Influenza A virus and its co infection with RSV among the hospitalized children with ARI. Methods: A total of 153 throat samples of the hospitalized young children aged between below one year and 5 years with the clinical signs of ARI were collected from the different hospitals in Khuzestan from June 2009 to April 2010. The samples were tested for Influenza A viruses by real time PCR. Positive IAV samples were tested for influenza A sub type H1N1 and for RSV by the nested PCR. Results: In this study, from the total 153 samples, 35 samples (22.9%) including 15 (42.8%) females and 20 (57.2%) males were positive for influenza A viruses. From the 35 positive samples for IAV, 14 were positive for swine H1N1 subtype. All the positive samples for influenza showed negative for RSV infection which revealed no coinfection with RSV. The prevalence of influenza A among age/sex groups was not significant. Conclusion: Influenza A is a prevalent viral agent isolated from young children with ARI. Influenza A subtype H1N1 was accounted for the 40 percent all laboratory-proven diagnoses of influenza in 2009. No evidence of coinfection of influenza A and RSV has been observed in the present study. PMID:24009929

Prevalence of Malaria, Dengue, and Chikungunya Significantly Associated with Mosquito Breeding Sites

PubMed Central

Islam, Mohammad Nazrul; ZulKifle, Mohammad; Sherwani, Arish Mohammad Khan; Ghosh, Susanta Kumar; Tiwari, Satyanarayan

2011-01-01

Objectives: To observe the prevalence of malaria, dengue, and chikungunya and their association with mosquito breeding sites. Methods: The study was observational and analytical. A total of 162 houses and 670 subjects were observed during the study period. One hundred forty-two febrile patients were eligible for the study. After obtaining informed consent from all febrile patients, 140 blood samples were collected to diagnose malaria, dengue, and chikungunya. Larval samples were collected by the standard protocol that follows. Correlation of data was performed by Pearson correlation test. Results: Forty-seven blood samples were found positive: 33 for chikungunya, 3 for dengue, and 11 for malaria. Fifty-one out of 224 larval samples were found positive. Out of the 51 positive samples, 37 were positive for Aedes, 12 were positive for Anopheles, and two were positive for Culex larvae. Interpretation and Conclusion: Mosquito-borne fevers, especially malaria, dengue, and chikungunya, have shown a significant relationship with mosquito breeding sites. PMID:23610486

Natural occurrence of aflatoxins and ochratoxin A in processed spices marketed in Malaysia.

PubMed

Ali, Norhayati; Hashim, Noor Hasani; Shuib, Nor Shifa

2015-01-01

The analysis of aflatoxins (B1, B2, G1 and G2) and ochratoxin A (OTA) was performed in processed spices marketed in Penang, Malaysia, using immunoaffinity columns and HPLC equipped with fluorescence detector (HPLC-FD). The processed powdered spices analysed include dried chilli, fennel, cumin, turmeric, black and white pepper, poppy seed, coriander, 'garam masala', and mixed spices for fish, meat and chicken curry. Two different studies were carried out. The limit of detection (LOD) was 0.01 ng g(-1) for each aflatoxin (AF) and 0.10 ng g(-1) for OTA (signal-to-noise ratio = 3:1). In the first study, 34 commercial processed spices analysed with a mean level, range and incidence of positive samples for total AF were 1.61 ng g(-1), 0.01-9.34 ng g(-1) and 85%, respectively, and for AFB1 were 1.38 ng g(-1), 0.01-7.68 ng g(-1) and 85%, respectively. The mean level, range and incidence of positive samples for OTA were 2.21 ng g(-1), 0.14-20.40 ng g(-1) and 79%, respectively. Natural co-occurrence of AF and OTA was found in 25 (74%) samples. In the second study of 24 commercial processed spices, the mean level, range and incidence of positive samples for total AF were 8.38 ng g(-1), 0.32-31.17 ng g(-1) and 88%, respectively, and for AFB1 were 7.31 ng g(-1), 0.32-28.43 ng g(-1) and 83%, respectively. Fifteen positive samples for total AF and two positive samples for OTA exceeded the permissible Malaysian limit of 5 ng g(-1). Contamination of both mycotoxins in spices may represent another route of exposure to consumers due to their frequent and prolonged consumption, as spices are common ingredients in popular dishes among Asian countries.

Incidence of Listeria spp. in Ready-to-Eat Food Processing Plant Environments Regulated by the U.S. Food Safety and Inspection Service and the U.S. Food and Drug Administration.

PubMed

Reinhard, Robert G; Kalinowski, Robin M; Bodnaruk, Peter W; Eifert, Joseph D; Boyer, Renee R; Duncan, Susan E; Bailey, R Hartford

2018-06-07

A multiyear survey of 31 ready-to-eat (RTE) food processing plants in the United States was conducted to determine the incidence of Listeria spp. in various RTE production environments. Samples were collected from 22 RTE plants regulated by the U.S. Department of Agriculture's Food Safety and Inspection Service (FSIS) and from 9 RTE food plants regulated by the U.S. Department of Health and Human Services' Food and Drug Administration (FDA). Only nonfood contact surfaces in the RTE manufacturing areas with exposed RTE product were sampled. Each sample was individually analyzed for the presence of Listeria spp. by using a PCR-based rapid assay. In total, 4,829 samples were collected from various locations, including freezers, equipment framework, floors, walls, wall-floor junctures, drains, floor mats, doors, and cleaning tools. Nine (29%) of the facilities had zero samples positive for Listeria spp. in the production environment, whereas 22 (71%) had one or more samples positive for Listeria spp. The total incidence of Listeria spp. in all RTE food plants was 4.5%. The positive rate in plants regulated by the FSIS ranged from 0 to 9.7%, whereas the positive rate in plants regulated by the FDA ranged from 1.2 to 36%.

Detection and zoonotic potential of Trichinella spp. from free-range pig farming in Greece.

PubMed

Papatsiros, V G; Boutsini, S; Ntousi, D; Stougiou, D; Mintza, D; Bisias, A

2012-06-01

Trichinellosis is a serious parasitic zoonosis, which is widely distributed around the world. Pork meat is still the predominant source of outbreaks of human trichinellosis in many countries. The aim of this study is to examine the impact of Trichinella spp. as an important risk factor on the free-range pig farming sector in Greece. In 2009, during routine testing for the detection of Trichinella larvae at slaughterhouses and the National Reference Laboratory for Parasites (NRL), a total of 826,426 pigs were tested with the magnetic stirrer method for Trichinella spp. at slaughterhouses, including 2,892 samples from free-range pigs. Two positive samples were detected: one positive for Trichinella britovi and one positive for Trichinella spp. (unspecified) in the samples from wild farmed free-range pigs. It is alarming that one of these cases was connected with clinical signs of trichinellosis in five persons of the same family in northeastern Greece, who consumed undercooked pork meat from a free-range pig farm. During 2010, a total number of 1,295,034 pigs were tested with same method, including 4,159 samples from free-range pig farms. Five positive samples for Trichinella spp. (unspecified) were detected from 4,159 free-range pigs tested by the Greek NRL. Moreover, 363 serum samples from free-range pigs were serologically tested with enzyme-linked immunosorbent assay (ELISA). Moreover, 363 serum samples from farmed free-range pigs were serologically tested with ELISA, and 15 samples were found positive. Finally, the present study is the first report of detection of T. britovi in Greece. In conclusion, based on the results of the present study, Trichinella spp. is a high-risk factor for the free-range pig farming in Greece.

Molecular surveillance of Theileria ovis, Theileria lestoquardi and Theileria annulata infection in sheep and ixodid ticks in Iran.

PubMed

Razmi, Gholamreza; Yaghfoori, Saeed

2013-01-01

A molecular study was undertaken to detect Theileria ovis, Theileria lestoquardi and Theileria annulata in sheep and tick vectors. Investigation was conducted from 2010 to 2011 in the south of Khorasan Razavi Province, Iran. A total of 150 blood samples were collected from 30 different sheep flocks. In addition, ixodid ticks were sampled from the same flocks. The stained blood smears were microscopically examined for the presence of piroplasms and a semi-nested polymerase chain reaction-restriction (PCR) was used for subsequent molecular speciation. Salivary glands were isolated from the ticks and subsequently analysed by semi-nested PCR. polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to differentiate between T. lestoquardi and T. annulata from PCR-positive samples. Theileria species infection was microscopically detected in 18.6% of blood smears. The presence of T. ovis and T. lestoquardi or T. annulata was detected by semi-nested PCR in 58.6% and 6.6% of blood samples respectively. In total, 169 ixodid ticks were collected from different areas of the province. The most prevalent ticks were Rhipicephalus turanicus (n = 155; 91.7% of the total), followed by Hyalomma anatolicum anatolicum (n = 8; 4.7%) and Hyalomma marginatum turanicum (n = 6; 3.5%). From an organ pooling of 33 ticks, three pools of salivary glands from R. turanicus were positive for Theileria species by semi-nested PCR. Of the three R. turanicus samples testing positive for Theileria species, two (6.1%) were positive for T. ovis and one (3.0%) for T. lestoquardi or T.annulata. Amongst the 11 PCR-positive samples for T. lestoquardi or T. annulata, 10 were positive for T. lestoquardi and one sample was positive for both T. lestoquardi and T. annulata using PCR-RFLP. The results also demonstrated that PCR-RFLP could be used for the detection of T. ovis. Based on the results, it can be concluded that T. ovis has a higher prevalence than T. lestoquardi, and that R. turanicus could be a possible vector for T. ovis and T. lestoquardi. Finally, the PCR-RFLP based on Msp1 restriction enzyme is a simple method for differentiation of Theileria species in sheep and ixodid ticks.

Solid fat content as a substitute for total polar compound analysis in edible oils

USDA-ARS?s Scientific Manuscript database

The solid fat contents (SFC) of heated edible oil samples were measured and found to correlate positively with total polar compounds (TPC) and inversely with triglyceride concentration. Traditional methods for determination of total polar compounds require a laboratory setting and are time intensiv...

Tube and column agglutination technology for autocontrol testing.

PubMed

Courtney, J E; Vincent, J L; Indrikovs, A J

2001-01-01

The incidence of positive autocontrol test results with column agglutination technology is a concern. This study investigates the incidence and significance of positive autocontrols in the ID Micro Typing System (gel) and the Gamma ReACT (ReACT). The study encompassed a total of 1021 randomly selected samples from patients and 95 samples from donors collected during 1 month. The autocontrol testing was carried out according to the manufacturer's instructions for the column agglutination tests. The tube method was carried out using low-ionic-strength solution (LISS). The direct antiglobulin test (DAT) was performed using the tube method, and further investigated with elution studies if warranted. Seventy-nine patient's samples (7.74%) had a positive autocontrol: the gel test, 72 (91.13%); ReACT, 21 (26.58%); and the tube method, 27 (34.18%). Of the 79 positive autocontrols, 44 samples had a negative DAT. Of the samples with positive DAT results, only one possessed a clinically significant antibody, anti-D. Moreover, the same sample also tested positive in all three methods. Column agglutination techniques have increased sensitivity for a positive autocontrol beyond the conventional tube method. However, ReACT and gel tests differ significantly in their frequency of positives. Investigation of the significance of a positive autocontrol in column agglutination technology when the conventional tube method is also positive is suggested.

Detection of Salmonella sp., Vibrio sp. and total plate count bacteria on blood cockle (Anadara granosa)

NASA Astrophysics Data System (ADS)

Ekawati, ER; Yusmiati, S. N. H.

2018-01-01

Blood cockle (Anadara granosa) has high level of zinc and protein, which is beneficial for therapeutic function for malnourished particularly stunting case in children. Zinc in animal foods is more absorbable than that from vegetable food. Blood cockle (Anadara granosa) is rich in nutrient and an excellent environment for the growth of microorganisms. This research aimed to identify the contamination of Salmonella sp., Vibrio sp. and total plate count bacteria on blood cockle (Anadara granosa). This was observation research with laboratory analysis. Salmonella sp. and Vibrio sp. were detected from blood cockle. Total plate count was determine of the total amount of the bacteria. Results detected from 20 samples of blood cockle showed that all samples were negative of Salmonella sp. and 1 sample positive Vibrio sp. The result of total plate count bacteria was < 5 x 105 colony/g sample.

Do we need to measure total serum IgA to exclude IgA deficiency in coeliac disease?

PubMed Central

Sinclair, D; Saas, M; Turk, A; Goble, M; Kerr, D

2006-01-01

Background Screening for IgA deficiency in patients with coeliac disease is essential because of the increased incidence of IgA deficiency associated with the disease, which usually relies on the estimation of IgA levels in each case. Aim To devise a method of excluding IgA deficiency without measuring total serum IgA in each case. Materials and methods The optical density readings on enzyme‐linked immunosorbent assay (ELISA) of 608 routine samples received for tissue transglutaminase (TTG) antibody testing for coeliac disease were compared with their total IgA concentrations. Dilution experiments were also carried out to ensure linear relationships between optical density on ELISA and IgA concentrations and to compare the sensitivities for TTG and endomysium antibodies in TTG‐positive samples. Results and discussion A clear relationship was shown between total IgA concentration and TTG optical density readings by ELISA. To ensure a positive TTG result if antibodies are present, it was possible to recommend an optical density level above which all samples have sufficient IgA. Samples with optical density <0.05 should be investigated further by estimating total IgA and, if low, samples should be subjected to immunofluorescence microscopy testing for IgA and IgG endomysium antibodies. Conclusions An easier, more cost‐effective and practical way of excluding IgA deficiency in the investigation on coeliac disease is reported. PMID:16489174

Associations between fecal indicator bacteria prevalence and demographic data in private water supplies in Virginia.

PubMed

Smith, Tamara; Krometis, Leigh-Anne H; Hagedorn, Charles; Lawrence, Annie H; Benham, Brian; Ling, Erin; Ziegler, Peter; Marmagas, Susan West

2014-12-01

Over 1.7 million Virginians rely on private water sources to provide household water. The heaviest reliance on these systems occurs in rural areas, which are often underserved with respect to available financial resources and access to environmental health education. This study aimed to identify potential associations between concentrations of fecal indicator bacteria (FIB) (coliforms, Escherichia coli) in over 800 samples collected at the point-of-use from homes with private water supply systems and homeowner-provided demographic data (household income and education). Of the 828 samples tested, 349 (42%) of samples tested positive for total coliform and 55 (6.6%) tested positive for E. coli. Source tracking efforts targeting optical brightener concentrations via fluorometry and the presence of a human-specific Bacteroides marker via quantitative real-time polymerase chain reaction (qPCR) suggest possible contamination from human septage in over 20 samples. Statistical methods implied that household income has an association with the proportion of samples positive for total coliform, though the relationship between education level and FIB is less clear. Further exploration of links between demographic data and private water quality will be helpful in building effective strategies to improve rural drinking water quality.

Easy fix for clinical laboratories for the false-positive defect with the Abbott AxSym total beta-hCG test.

PubMed

Cole, Laurence A; Khanlian, Sarah A

2004-05-01

False-positive hCG results can lead to erroneous diagnoses and needless chemotherapy and surgery. In the last 2 years, eight publications described cases involving false-positive hCG tests; all eight involved the AxSym test. We investigated the source of this abundance of cases and a simple fix that may be used by clinical laboratories. False-positive hCG was primarily identified by absence of hCG in urine and varying or negative hCG results in alternative tests. Seventeen false-positive serum samples in the AxSym test were evaluated undiluted and at twofold dilution with diluent containing excess goat serum or immunoglobulin. We identified 58 patients with false-positive hCG, 47 of 58 due to the Abbott AxSym total hCGbeta test (81%). Sixteen of 17 of these "false-positive" results (mean 100 mIU/ml) became undetectable when tested again after twofold dilution. A simple twofold dilution with this diluent containing excess goat serum or immunoglobulin completely protected 16 of 17 samples from patients having false-positive results. It is recommended that laboratories using this test use twofold dilution as a minimum to prevent false-positive results.

Opportunistic parasites among immunosuppressed children in Minia District, Egypt.

PubMed

Abdel-Hafeez, Ekhlas H; Ahmad, Azza K; Ali, Basma A; Moslam, Fadia A

2012-03-01

A total of 450 stool samples were collected from inpatient and outpatient clinics of Pediatric Department, Minia University Hospital, Minia District, Egypt. Two groups of patients were studied, including 200 immunosuppressed and 250 immunocompetent children. Stool samples were subjected to wet saline and iodine mounts. A concentration technique (formol-ether sedimentation method) was carried out for stool samples diagnosed negative by wet saline and iodine mounts. Samples were stained by 2 different methods; acid fast stain (modified Ziehl-Neelsen stain) and Giemsa stain. Total 188 cases (94%) were diagnosed positive for parasitic infections among immunosuppressed children, whereas 150 cases (60%) were positive in immunocompetent children (P<0.0001). The most common protozoan infection in immunosuppressed group was Cryptosporidium parvum (60.2%), followed by Blastocystis hominis (12.1%), Isospora belli (9.7%), and Cyclospora caytenensis (7.8%). On the other hand, Entamoeba histolytica (24.6%) and Giardia lamblia (17.6%) were more common than other protozoans in immunocompetent children.

[Study on tetrodotoxin detection and toxic puffer fish identification of roasted fish fillet at the retail in Beijing and Qingdao].

PubMed

Shen, Qing; Jiang, Tao; Li, Nan; Wang, Jiahui; Han, Chunhui; Zhang, Jing; Xu, Jin; Zhang, Dongfeng; Li, Fengqin

2014-11-01

The roasted fish fillet sample at the retail collected in Beijing and Qingdao were detected for TTX, and the TTX positive samples was analyzed for fish species identification. TTX was tested by EUSA method and the cytochrome c oxidase I (COI) genome of TTX-positive samples was extracted and identified by DNA barcode. Totally, 90 samples were tested by EUSA and 58 (64.4%) samples were positive for TTX with the levels ranging from 0.10 mg/kg to 63.81 mg/kg. Among the TTX positive samples, 24 (41.3%) were identified containing toxic puffer fish and 21 (87.5%) were Lagocephalus lunaris, the highly toxic puffer fish. Some roasted fish fillet samples obtained from the retail in two cities were positive for TTX and contained toxic puffer fish. Based on these results, we suggest that roasted fish fillet producers should prevent toxic puffer fish from mixing in the raw material and the I regulators should strengthen the TTX surveillance and product labeling supervision of roasted fish fillet.

Short communication: Microbiological quality of raw cow milk and its association with herd management practices in Northern China.

PubMed

Lan, X Y; Zhao, S G; Zheng, N; Li, S L; Zhang, Y D; Liu, H M; McKillip, J; Wang, J Q

2017-06-01

Contamination of raw milk with bacterial pathogens is potentially hazardous to human health. The aim of this study was to evaluate the total bacteria count (TBC) and presence of pathogens in raw milk in Northern China along with the associated herd management practices. A total of 160 raw milk samples were collected from 80 dairy herds in Northern China. All raw milk samples were analyzed for TBC and pathogens by culturing. The results showed that the number of raw milk samples with TBC <2 × 10 6 cfu/mL and <1 × 10 5 cfu/mL was 146 (91.25%) and 70 (43.75%), respectively. A total of 84 (52.50%) raw milk samples were Staphylococcus aureus positive, 72 (45.00%) were Escherichia coli positive, 2 (1.25%) were Salmonella positive, 2 (1.25%) were Listeria monocytogenes positive, and 3 (1.88%) were Campylobacter positive. The prevalence of S. aureus was influenced by season, herd size, milking frequency, disinfection frequency, and use of a Dairy Herd Improvement program. The TBC was influenced by season and milk frequency. The correlation between TBC and prevalence of S. aureus or E. coli is significant. The effect size statistical analysis showed that season and herd (but not Dairy Herd Improvement, herd size, milking frequency, disinfection frequency, and area) were the most important factors affecting TBC in raw milk. In conclusion, the presence of bacteria in raw milk was associated with season and herd management practices, and further comprehensive study will be powerful for effectively characterizing various factors affecting milk microbial quality in bulk tanks in China. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Legionella species colonization of water distribution systems, pools and air conditioning systems in cruise ships and ferries

PubMed Central

Goutziana, Georgia; Mouchtouri, Varvara A; Karanika, Maria; Kavagias, Antonios; Stathakis, Nikolaos E; Gourgoulianis, Kostantinos; Kremastinou, Jenny; Hadjichristodoulou, Christos

2008-01-01

Background Legionnaires' disease continues to be a public health concern in passenger ships. This study was scheduled in order to investigate Legionella spp. colonization of water distribution systems (WDS), recreational pools, and air-conditioning systems on board ferries and cruise ships in an attempt to identify risk factors for Legionella spp. colonization associated with ship water systems and water characteristics. Methods Water systems of 21 ferries and 10 cruise ships including WDS, air conditioning systems and pools were investigated for the presence of Legionella spp. Results The 133 samples collected from the 10 cruise ships WDS, air conditioning systems and pools were negative for Legionella spp. Of the 21 ferries WDS examined, 14 (66.7%) were legionellae-positive. A total of 276 samples were collected from WDS and air conditioning systems. Legionella spp. was isolated from 37.8% of the hot water samples and 17.5% of the cold water samples. Of the total 96 positive isolates, 87 (90.6%) were L. pneumophila. Legionella spp. colonization was positively associated with ship age. The temperature of the hot water samples was negatively associated with colonization of L. pneumophila serogroup (sg) 1 and that of L. pneumophila sg 2 to 14. Increases in pH ≥7.8 and total plate count ≥400 CFU/L, correlated positively with the counts of L. pneumophila sg 2 to 14 and Legionella spp. respectively. Free chlorine of ≥0.2 mg/L inhibited colonization of Legionella spp. Conclusion WDS of ferries can be heavily colonized by Legionella spp. and may present a risk of Legionnaires' disease for passengers and crew members. Guidelines and advising of Legionnaires' disease prevention regarding ferries are needed, in particular for operators and crew members. PMID:19025638

Legionella species colonization of water distribution systems, pools and air conditioning systems in cruise ships and ferries.

PubMed

Goutziana, Georgia; Mouchtouri, Varvara A; Karanika, Maria; Kavagias, Antonios; Stathakis, Nikolaos E; Gourgoulianis, Kostantinos; Kremastinou, Jenny; Hadjichristodoulou, Christos

2008-11-24

Legionnaires' disease continues to be a public health concern in passenger ships. This study was scheduled in order to investigate Legionella spp. colonization of water distribution systems (WDS), recreational pools, and air-conditioning systems on board ferries and cruise ships in an attempt to identify risk factors for Legionella spp. colonization associated with ship water systems and water characteristics. Water systems of 21 ferries and 10 cruise ships including WDS, air conditioning systems and pools were investigated for the presence of Legionella spp. The 133 samples collected from the 10 cruise ships WDS, air conditioning systems and pools were negative for Legionella spp. Of the 21 ferries WDS examined, 14 (66.7%) were legionellae-positive. A total of 276 samples were collected from WDS and air conditioning systems. Legionella spp. was isolated from 37.8% of the hot water samples and 17.5% of the cold water samples. Of the total 96 positive isolates, 87 (90.6%) were L. pneumophila. Legionella spp. colonization was positively associated with ship age. The temperature of the hot water samples was negatively associated with colonization of L. pneumophila serogroup (sg) 1 and that of L. pneumophila sg 2 to 14. Increases in pH >/=7.8 and total plate count > or =400 CFU/L, correlated positively with the counts of L. pneumophila sg 2 to 14 and Legionella spp. respectively. Free chlorine of > or =0.2 mg/L inhibited colonization of Legionella spp. WDS of ferries can be heavily colonized by Legionella spp. and may present a risk of Legionnaires' disease for passengers and crew members. Guidelines and advising of Legionnaires' disease prevention regarding ferries are needed, in particular for operators and crew members.

How Many Cultures Are Necessary to Identify Pathogens in the Management of Total Hip and Knee Arthroplasty Infections?

PubMed

Gandhi, Rikesh; Silverman, Edward; Courtney, Paul M; Lee, Gwo-Chin

2017-09-01

Identification of the infecting organism is critical to the successful management of deep prosthetic joint infections about the hip and the knee. However, the number of culture specimens and which culture specimens are best to identify these organisms is unknown. We evaluated 113 consecutive patients with infected total hip and total knee arthroplasties and correlated the type of culture specimen and number of specimens taken during surgery to the likelihood of a positive culture result. From these data, we subsequently developed a model to maximize culture yield at the time of surgical intervention. After exclusions, 74 patients meeting the Musculoskeletal Infection Society criteria were left for final analysis. From this cohort, 63 of 74 patients had a positive culture result (85%). The odds of a fluid culture result being positive was 35 of 47 (0.75), whereas the likelihood of tissue cultures yielding a positive result was 164 of 245 (0.67; P = .313). The sample designated "best culture" specimen was the only culture with a positive result in 1 of 48 cases in which a best culture was identified. The optimal number of cultures needed to yield a positive test result was 4 (specificity = 0.61 and sensitivity = 0.63). Increasing the number of samples increases sensitivity but reduces specificity. A minimum of 4 tissue cultures from representative areas is necessary to maximize the chance of identifying the infecting organism during management of the infected total hip and total knee arthroplasties. The designation of the best culture specimen for additional testing is arbitrary and may not be clinically efficacious. Copyright © 2017 Elsevier Inc. All rights reserved.

Surveillance of Total Mercury and Methylmercury Concentrations in Retail Fish.

PubMed

Watanabe, Takahiro; Hayashi, Tomoko; Matsuda, Rieko; Akiyama, Hiroshi; Teshima, Reiko

2017-01-01

Most fish samples contain methylmercury, that the concentrations very greatly according to the fish species. To avoid the adverse health effects of methylmercury while retaining the benefits provided by fish consumption, it is important to select suitable fish species and to control the amount of the fish intake. We surveyed the concentrations of total mercury and methylmercury in 210 retail fish samples classified into 19 fish species by using validated analytical methods. The results of this survey were as follows. The total mercury and methylmercury concentrations were higher than 1 mg/kg in some samples of swordfish and bluefin tuna, which are large predatory fish species. In bluefin tuna and yellowtail, total mercury and methylmercury concentrations in farm-raised fish were lower than those in natural fish. There was a positive correlation between total mercury concentration and methylmercury concentration. Our results indicate that a cut-off value of 0.3 mg/kg total mercury in the screening of fish samples would increase the effectiveness of inspection.

17 CFR Appendix B to Part 420 - Sample Large Position Report

Code of Federal Regulations, 2013 CFR

2013-04-01

... $ Memorandum 2: Report the gross par amount of fails to deliver. Included in the calculation of line item 3... millions at par value as of trade date] Security Being Reported Date For Which Information is Being... Principal Components of the Specific Security $ Total Net Trading Position $ 2. Gross Financing Position...

17 CFR Appendix B to Part 420 - Sample Large Position Report

Code of Federal Regulations, 2012 CFR

2012-04-01

... $ Memorandum 2: Report the gross par amount of fails to deliver. Included in the calculation of line item 3... millions at par value as of trade date] Security Being Reported Date For Which Information is Being... Principal Components of the Specific Security $ Total Net Trading Position $ 2. Gross Financing Position...

17 CFR Appendix B to Part 420 - Sample Large Position Report

Code of Federal Regulations, 2011 CFR

2011-04-01

... $ Memorandum 2: Report the gross par amount of fails to deliver. Included in the calculation of line item 3... millions at par value as of trade date] Security Being Reported Date For Which Information is Being... Principal Components of the Specific Security $ Total Net Trading Position $ 2. Gross Financing Position...

Reemergence of yellow fever: detection of transmission in the State of São Paulo, Brazil, 2008.

PubMed

Moreno, Eduardo Stramandinoli; Rocco, Iray Maria; Bergo, Eduardo Sterlino; Brasil, Roosecelis Araujo; Siciliano, Melissa Mascheratti; Suzuki, Akemi; Silveira, Vivian Regina; Bisordi, Ivani; Souza, Renato Pereira de

2011-01-01

Following yellow fever virus (YFV) isolation in monkeys from the São José do Rio Preto region and two fatal human autochthonous cases from the Ribeirão Preto region, State of São Paulo, Brazil, two expeditions for entomological research and eco-epidemiological evaluation were conducted. A total of 577 samples from humans, 108 from monkeys and 3,049 mosquitoes were analyzed by one or more methods: virus isolation, ELISA-IgM, RT-PCR, histopathology and immunohistochemical. Of the 577 human samples, 531 were tested by ELISA-IgM, with 3 positives, and 235 were inoculated into mice and 199 in cell culture, resulting in one virus isolation. One sample was positive by histopathology and immunohistochemical. Using RT-PCR, 25 samples were processed with 4 positive reactions. A total of 108 specimens of monkeys were examined, 108 were inoculated into mice and 45 in cell culture. Four virus strains were isolated from Alouatta caraya. A total of 931 mosquitoes were captured in Sao Jose do Rio Preto and 2,118 in Ribeirão Preto and separated into batches. A single isolation of YFV was derived from a batch of 9 mosquitoes Psorophora ferox, collected in Urupês, Ribeirão Preto region. A serological survey was conducted with 128 samples from the municipalities of São Carlos, Rincão and Ribeirão Preto and 10 samples from contacts of patients from Ribeirão Preto. All samples were negative by ELISA-IgM for YFV. The results confirm the circulation of yellow fever, even though sporadic, in the Sao Paulo State and reinforce the importance of vaccination against yellow fever in areas considered at risk.

Impact of phlebotomine sand flies on U.S. military operations at Tallil Air Base, Iraq: 4. Detection and identification of leishmania parasites in sand flies.

PubMed

Coleman, Russell E; Hochberg, Lisa P; Swanson, Katherine I; Lee, John S; McAvin, James C; Moulton, John K; Eddington, David O; Groebner, Jennifer L; O'Guinn, Monica L; Putnam, John L

2009-05-01

Sand flies collected between April 2003 and November 2004 at Tallil Air Base, Iraq, were evaluated for the presence of Leishmania parasites using a combination of a real-time Leishmania-generic polymerase chain reaction (PCR) assay and sequencing of a 360-bp fragment of the glucose-6-phosphate-isomerase (GPI) gene. A total of 2,505 pools containing 26,574 sand flies were tested using the real-time PCR assay. Leishmania DNA was initially detected in 536 pools; however, after extensive retesting with the real-time PCR assay, a total of 456 pools were considered positive and 80 were considered indeterminate. A total of 532 samples were evaluated for Leishmania GPI by sequencing, to include 439 PCR-positive samples, 80 PCR-indeterminate samples, and 13 PCR-negative samples. Leishmania GPI was detected in 284 samples that were sequenced, to include 281 (64%) of the PCR-positive samples and 3 (4%) of the PCR-indeterminate samples. Of the 284 sequences identified as Leishmania, 261 (91.9%) were L. tarentolae, 18 (6.3%) were L. donovani-complex parasites, 3 (1.1%) were L. tropica, and 2 were similar to both L. major and L. tropica. Minimum field infection rates were 0.09% for L. donovani-complex parasites, 0.02% for L. tropica, and 0.01% for the L. major/tropica-like parasite. Subsequent sequencing of a 600-bp region of the "Hyper" gene of 12 of the L. donovani-complex parasites showed that all 12 parasites were L. infantum. These data suggest that L. infantum was the primary leishmanial threat to U.S. military personnel deployed to Tallil Air Base. The implications of these findings are discussed.

Bacteriology of Dehydrated Space Foods 1

PubMed Central

Powers, Edmund M.; Ay, Carl; El-Bisi, Hamed M.; Rowley, Durwood B.

1971-01-01

The initial bacteriological requirement established in 1964 for space foods by the U.S. Army Natick Laboratories are: a total aerobic plate count (≤ 10,000 per g), a total coliform count (≤ 10 per g), fecal coliforms (negative per gram), fecal streptococci (≤ 20 per g), coagulase-positive staphylococci (negative in 5 g) and salmonellae (negative in 10 g). Of the space foods and prototypes tested during 1968 and 1969, 93% complied with the total aerobic plate count, 98% had less than 1 coliform per g, and 99% were negative for fecal coliforms; 88% complied with the streptococci requirement; 100 and 98% were negative for staphylococci and salmonellae, respectively. Nineteen food samples which did not comply (as indicated parenthetically by actual counts per gram) with the requirements were (i) total aerobic plate count: beef soup and gravy base (18,000), chicken soup and gravy base (57,000), spaghetti with meat sauce (12,100 and 14,000), sugared coffee (> 300,000), chocolate ice cream cubes (20,000), and each of four samples of chocolate candy (12,000 to 61,000); (ii) coliforms: two out of three vanilla milk drinks (16 and 127) and one beef hash bar (14); (iii) fecal coliforms: one sample of chicken soup and gravy base positive; (iv) fecal streptococci: two samples of peanut cubes (40 and 108), coconut cubes (75), chicken soup and gravy base (2,650), beef soup and gravy base (33), and five out of six flavored milk drinks (23 to 300); (v) salmonellae: one each of chicken and beef soup and gravy base were positive. Images PMID:4940878

A field survey using LAMP assay for detection of Schistosoma mansoni in a low-transmission area of schistosomiasis in Umbuzeiro, Brazil: Assessment in human and snail samples.

PubMed

Gandasegui, Javier; Fernández-Soto, Pedro; Muro, Antonio; Simões Barbosa, Constança; Lopes de Melo, Fabio; Loyo, Rodrigo; de Souza Gomes, Elainne Christine

2018-03-01

In Brazil, schistosomiasis is a parasitic disease of public health relevance, mainly in poor areas where Schistosoma mansoni is the only human species encountered and Biomphalaria straminea is one of the intermediate host snails. A nested-PCR based on a specific mitochondrial S. mansoni minisatellite DNA region has been successfully developed and applied as a reference method in Brazil for S. mansoni detection, mainly in host snails for epidemiological studies. The amplification efficiency of LAMP is known to be higher than PCR. The present work aimed to assess the utility of our previously described SmMIT-LAMP assay for S. mansoni detection in human stool and snail samples in a low-transmission area of schistosomiasis in the municipality of Umbuzeiro, Paraíba State, Northeast Region of Brazil. A total of 427 human stool samples were collected during June-July 2016 in the municipality of Umbuzeiro and an overall prevalence of 3.04% (13/427) resulted positive by duplicate Kato-Katz thick smear. A total of 1,175 snails identified as Biomphalaria straminea were collected from 14 breeding sites along the Paraíba riverbank and distributed in 46 pools. DNA from human stool samples and pooled snails was extracted using the phenol/chloroform method. When performing the SmMIT-LAMP assay a total of 49/162 (30.24%) stool samples resulted positive, including 12/13 (92.31%) that were Kato-Katz positive and 37/149 (24.83%) previously Kato-Katz negative. By nested-PCR, only 1/46 pooled DNA snail samples was positive. By SmMIT-LAMP assay, the same sample also resulted positive and an additional one was positive from a different breeding site. Data of human and snail surveys were used to build risk maps of schistosomiasis incidence using kernel density analysis. This is the first study in which a LAMP assay was evaluated in both human stool and snail samples from a low-transmission schistosomiasis-endemic area. Our SmMIT-LAMP proved to be much more efficient in detection of S. mansoni in comparison to the 'gold standard' Kato-Katz method in human stool samples and the reference molecular nested-PCR in snails. The SmMIT-LAMP has demonstrated to be a useful molecular tool to identify potential foci of transmission in order to build risk maps of schistosomiasis.

Efficiently estimating salmon escapement uncertainty using systematically sampled data

USGS Publications Warehouse

Reynolds, Joel H.; Woody, Carol Ann; Gove, Nancy E.; Fair, Lowell F.

2007-01-01

Fish escapement is generally monitored using nonreplicated systematic sampling designs (e.g., via visual counts from towers or hydroacoustic counts). These sampling designs support a variety of methods for estimating the variance of the total escapement. Unfortunately, all the methods give biased results, with the magnitude of the bias being determined by the underlying process patterns. Fish escapement commonly exhibits positive autocorrelation and nonlinear patterns, such as diurnal and seasonal patterns. For these patterns, poor choice of variance estimator can needlessly increase the uncertainty managers have to deal with in sustaining fish populations. We illustrate the effect of sampling design and variance estimator choice on variance estimates of total escapement for anadromous salmonids from systematic samples of fish passage. Using simulated tower counts of sockeye salmon Oncorhynchus nerka escapement on the Kvichak River, Alaska, five variance estimators for nonreplicated systematic samples were compared to determine the least biased. Using the least biased variance estimator, four confidence interval estimators were compared for expected coverage and mean interval width. Finally, five systematic sampling designs were compared to determine the design giving the smallest average variance estimate for total annual escapement. For nonreplicated systematic samples of fish escapement, all variance estimators were positively biased. Compared to the other estimators, the least biased estimator reduced bias by, on average, from 12% to 98%. All confidence intervals gave effectively identical results. Replicated systematic sampling designs consistently provided the smallest average estimated variance among those compared.

Comparison of CpG island methylator phenotype (CIMP) frequency in colon cancer using different probe- and gene-specific scoring alternatives on recommended multi-gene panels.

PubMed

Berg, Marianne; Hagland, Hanne R; Søreide, Kjetil

2014-01-01

In colorectal cancer a distinct subgroup of tumours demonstrate the CpG island methylator phenotype (CIMP). However, a consensus of how to score CIMP is not reached, and variation in definition may influence the reported CIMP prevalence in tumours. Thus, we sought to compare currently suggested definitions and cut-offs for methylation markers and how they influence CIMP classification in colon cancer. Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA), with subsequent fragment analysis, was used to investigate methylation of tumour samples. In total, 31 CpG sites, located in 8 different genes (RUNX3, MLH1, NEUROG1, CDKN2A, IGF2, CRABP1, SOCS1 and CACNA1G) were investigated in 64 distinct colon cancers and 2 colon cancer cell lines. The Ogino gene panel includes all 8 genes, in addition to the Weisenberger panel of which only 5 of the 8 genes included were investigated. In total, 18 alternative combinations of scoring of CIMP positivity on probe-, gene-, and panel-level were analysed and compared. For 47 samples (71%), the CIMP status was constant and independent of criteria used for scoring; 34 samples were constantly scored as CIMP negative, and 13 (20%) consistently scored as CIMP positive. Only four of 31 probes (13%) investigated showed no difference in the numbers of positive samples using the different cut-offs. Within the panels a trend was observed that increasing the gene-level stringency resulted in a larger difference in CIMP positive samples than increasing the probe-level stringency. A significant difference between positive samples using 'the most stringent' as compared to 'the least stringent' criteria (20% vs 46%, respectively; p<0.005) was demonstrated. A statistical significant variation in the frequency of CIMP depending on the cut-offs and genes included in a panel was found, with twice as many positives samples by least compared to most stringent definition used.

Groundwater quality assessment in the village of Lutfullapur Nawada, Loni, District Ghaziabad, Uttar Pradesh, India.

PubMed

Singh, Vinod K; Bikundia, Devendra Singh; Sarswat, Ankur; Mohan, Dinesh

2012-07-01

The groundwater quality for drinking, domestic and irrigation in the village Lutfullapur Nawada, Loni, district Ghaziabad, U.P., India, has been assessed. Groundwater samples were collected, processed and analyzed for temperature, pH, conductivity, salinity, total alkalinity, carbonate alkalinity, bicarbonate alkalinity, total hardness, calcium hardness, magnesium hardness, total solids, total dissolved solids, total suspended solids, nitrate-nitrogen, chloride, fluoride, sulfate, phosphate, silica, sodium, potassium, calcium, magnesium, total chromium, cadmium, copper, iron, nickel, lead and zinc. A number of groundwater samples showed levels of electrical conductivity (EC), alkalinity, chloride, calcium, sodium, potassium and iron exceeding their permissible limits. Except iron, the other metals (Cr, Cd, Cu, Ni, Pb, and Zn) were analyzed below the permissible limits. The correlation matrices for 28 variables were performed. EC, salinity, TS and TDS had significant positive correlations among themselves and also with NO (3) (-) , Cl(-), alkalinity, Na(+), K(+), and Ca(2+). Fluoride was not significantly correlated with any of the parameters. NO (3) (-) was significantly positively correlated with Cl(-), alkalinity, Na(+), K(+) and Ca(2+). Chloride also correlated significantly with alkalinity, Na(+), K(+) and Ca(2+). Sodium showed a strong and positive correlation with K(+) and Ca(2+). pH was negatively correlated with most of the physicochemical parameters. This groundwater is classified as a normal sulfate and chloride type. Base-exchange indices classified 73% of the groundwater sources as the Na(+)-SO (4) (2-) type. The meteoric genesis indices demonstrated that 67% of groundwater sources belong to a deep meteoric water percolation type. Hydrochemical groundwater evaluations revealed that most of the groundwaters belong to the Na(+)-K(+)-Cl(-)-SO (4) (2-) type followed by Na(+)-K(+)-HCO (3) (-) type. Salinity, chlorinity and SAR indices indicated that majority of groundwater samples can be considered suitable for irrigation purposes.

Detection of Toxoplasma gondii DNA in Brazilian oysters (Crassostrea rhizophorae).

PubMed

Ribeiro, L A; Santos, L K N S S; Brito, P A; Maciel, B M; Da Silva, A V; Albuquerque, G R

2015-05-04

The aim of this study was to detect evidence of Toxoplasma gondii using polymerase chain reaction (PCR)-based techniques in oysters (Crassostrea rhizophorae) obtained from the southern coastal region of Bahia, Brazil. A total of 624 oysters were collected, and the gills and digestive glands were dissected. Each tissue sample was separated into pools containing tissues (of the same type) from three animals, leading to a total of 416 experimental samples for analysis (208 samples each from the gills and digestive glands). Molecular analysis using PCR-based detection of the T. gondii AF 146527 repetitive fragment yielded negative results for all samples. However, when nested-PCR was used for detection of the T. gondii SAG-1 gene, 17 samples were positive, with the gills being the tissue with maximal detection of the parasite. These positive results were confirmed by sample sequencing. It is therefore suggested that C. rhizophorae oysters are capable of filtering and retaining T. gondii oocysts in their tissue. This represents a risk to public health because they are traditionally ingested in natura.

Total coliform and Escherichia coli contamination in rural well water: analysis for passive surveillance.

PubMed

Invik, Jesse; Barkema, Herman W; Massolo, Alessandro; Neumann, Norman F; Checkley, Sylvia

2017-10-01

With increasing stress on our water resources and recent waterborne disease outbreaks, understanding the epidemiology of waterborne pathogens is crucial to build surveillance systems. The purpose of this study was to explore techniques for describing microbial water quality in rural drinking water wells, based on spatiotemporal analysis, time series analysis and relative risk mapping. Tests results for Escherichia coli and coliforms from private and small public well water samples, collected between 2004 and 2012 in Alberta, Canada, were used for the analysis. Overall, 14.6 and 1.5% of the wells were total coliform and E. coli-positive, respectively. Private well samples were more often total coliform or E. coli-positive compared with untreated public well samples. Using relative risk mapping we were able to identify areas of higher risk for bacterial contamination of groundwater in the province not previously identified. Incorporation of time series analysis demonstrated peak contamination occurring for E. coli in July and a later peak for total coliforms in September, suggesting a temporal dissociation between these indicators in terms of groundwater quality, and highlighting the potential need to increase monitoring during certain periods of the year.

Detection of Mycobacterium avium subspecies paratuberculosis specific IS900 insertion sequences in bulk-tank milk samples obtained from different regions throughout Switzerland

PubMed Central

Corti, Sabrina; Stephan, Roger

2002-01-01

Background Since Mycobacterium avium subspecies paratuberculosis (MAP) was isolated from intestinal tissue of a human patient suffering Crohn's disease, a controversial discussion exists whether MAP have a role in the etiology of Crohn's disease or not. Raw milk may be a potential vehicle for the transmission of MAP to human population. In a previous paper, we have demonstrated that MAP are found in raw milk samples obtained from a defined region in Switzerland. The aim of this work is to collect data about the prevalence of MAP specific IS900 insertion sequence in bulk-tank milk samples in different regions of Switzerland. Furthermore, we examined eventual correlation between the presence of MAP and the somatic cell counts, the total colony counts and the presence of Enterobacteriaceae. Results 273 (19.7%) of the 1384 examined bulk-tank milk samples tested IS900 PCR-positive. The prevalence, however, in the different regions of Switzerland shows significant differences and ranged from 1.7% to 49.2%. Furthermore, there were no statistically significant (p >> 0.05) differences between the somatic cell counts and the total colony counts of PCR-positive and PCR-negative milk samples. Enterobacteriaceae occur as often in IS900 PCR-positive as in PCR-negative milk samples. Conclusion This is the first study, which investigates the prevalence of MAP in bulk-tank milk samples all over Switzerland and infers the herd-level prevalence of MAP infection in dairy herds. The prevalence of 19.7% IS900 PCR-positive bulk-milk samples shows a wide distribution of subclinical MAP-infections in dairy stock in Switzerland. MAP can therefore often be transmitted to humans by raw milk consumption. PMID:12097144

[Identification of Vibrio cholerae O1 by flow cytometry].

PubMed

Alvarado-Alemán, F J; González-Bonilla, C; Wong-Arambula, C; Gutiérrez-Cogco, L; Sepúlveda-Amor, J; Kumate-Rodríguez, J

1994-01-01

A total of 72 peptonated water samples suspected of carrying Vibrio cholerae were assessed by laser flow cytometry (LFC) and compared with positive culture. We used a direct fluorescence technique using polyclonal (PolAb) and monoclonal antibodies (MoAb) conjugated to fluorescein. The PolAb were able to detect 33 positive samples. A clear difference among the 20 positive samples was found with only three V. cholerae O1 false negatives when MoAb were used whereas all 13 V. cholerae Non O1 samples were detected. The correlation index comparing control autofluorescence with peptonated water samples show a R = 0.69, versus 0.96 with pure V. cholerae O1 strains. Our data suggest that the LFC technique is able to recognize V. cholerae O1 from a mixture of microorganisms with high sensitivity and specificity in a few hours.

Microbiological quality of fresh nopal juice

USDA-ARS?s Scientific Manuscript database

Microbial quality of commercially available fresh juices (n=162) made with nopal cactus in Texcoco, State of Mexico, was evaluated during the summer and spring season. All samples contained total coliforms and 91% were positive for Escherichia coli. Although total coliforms and E. coli were detected...

Prevalence, types, and geographical distribution of Listeria monocytogenes from a survey of retail Queso Fresco and associated cheese processing plants and dairy farms in Sonora, Mexico.

PubMed

Moreno-Enriquez, R I; Garcia-Galaz, A; Acedo-Felix, E; Gonzalez-Rios, I H; Call, J E; Luchansky, J B; Diaz-Cinco, M E

2007-11-01

In the first part of this study, samples were collected from farms, cheese processing plants (CPPs), and retail markets located in various geographical areas of Sonora, Mexico, over a 12-month period during the summer of 2004 and winter of 2005. Four (all Queso Fresco [QF] from retail markets) of 349 total samples tested positive for Listeria monocytogenes (Lm). Of these four positive samples, three were collected in the northern region and one in the southern region of Sonora. Additionally, two were collected during the winter months, and two were collected during the summer months. For the second part of the study, a total of 39 samples from a farm, a CPP, and retail markets were collected and processed according to a combination of the Norma Oficial Mexicana NOM-143-SSA1-1995.10 method (NOM) and the U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual method, and 27 samples from these same locations were collected and processed according to the U.S. Department of Agriculture Food Safety and Inspection Service method (USDA-FSIS). The NOM-FDA method recovered the pathogen from 6 (15%) of 39 samples (one cheese and five product contact surfaces), while the USDA-FSIS method recovered the pathogen from 5 (18.5%) of 27 samples (all product contact surfaces). In addition, the 40 isolates recovered from the 15 total samples that tested positive for Lm grouped into five distinct pulsotypes that were ca. 60% related, as determined by pulsed-field gel electrophoresis analysis. The results of this study confirmed a 3.4% prevalence of Lm in QF collected from retail markets located in Sonora and no appreciable difference in the effectiveness of either the NOM-FDA or USDA-FSIS method to recover the pathogen from cheese or environmental samples.

Laboratory investigation of drinking water sources of Kangra, Himachal Pradesh.

PubMed

Thakur, S D; Panda, A K

2012-06-01

A total of 70 drinking water sources including piped water supply (n = 36), ground water sources (n = 24, hand pumps and bore wells) and natural water sources (n = 10, springs/step-wells) from various parts of district Kangra, Himachal Pradesh were investigated for their suitability for drinking purpose by presumptive coliform test. Three samples were collected from each source during different parts of the year. Piped water sources (91.7%) were most contaminated followed by natural water sources (90%) and ground water sources (62.5%). 70.5% of the total water samples (n = 210) were positive for coliforms. All the three samples from 8.3% (n = 3), 37.5% (n = 9) and 10% (n = 1) piped water, ground water and natural sources respectively, were negative for coliform organisms. A variety of organisms including Proteus, Klebsiella, Citrobacter, Escherichia coli (E. coli), Pasteurella, Enterobacter and Serratia liquefaciens were isolated from water samples positive for coliforms in presumptive coliform test. Thermo-tolerant coliform organisms; Escherichia coli, Citrobacter, Klebsiella and Enterobacter were 71.2% (n = 52) of the total bacterial isolations. These findings suggest absence of adequate treatment and disinfection of the water sources supplying drinking water in district Kangra.

Assessing Trophic Position and Mercury Accumulation in Sanpping Turtles

EPA Science Inventory

This study determined the trophic position and the total mercury concentrations of snapping turtles (Chelydra serpentina) captured from 26 freshwater sites in Rhode Island. Turtles were captured in baited wire cages, and a non-lethal sampling technique was used in which tips of ...

Comparison of the incidence of Listeria on equipment versus environmental sites within dairy processing plants.

PubMed

Pritchard, T J; Flanders, K J; Donnelly, C W

1995-08-01

This study was undertaken to compare the incidence of Listeria contamination of processing equipment with that of the general dairy processing environment. A total of 378 sponge samples obtained from 21 dairy plants were analyzed for Listeria using three different enrichment media. Use of extended microbiological analysis allowed us to identify 26 Listeria positive sites which would have not been identified had a single test format been employed. Eighty (80) of 378 sites (21.2%) were identified as Listeria positive. Listeria innocua was isolated from 59 of the 80 (73.8%) positive samples, L. monocytogenes was identified in 35 (43.8%) of the positive samples, and L. seeligeri was isolated from 5 (6.3%) of the Listeria positive samples. Positive equipment samples were obtained from 6 of the 21 (28.6%) plants and 19 of the 21 (90.5%) plants had positive environmental sites. Seventeen of the 215 (7.9%) samples from equipment were positive for Listeria species. Eleven of these sites, including 3 holding tanks, 2 table tops, 3 conveyor/chain systems, a pasta filata wheel, a pint milk filler and a brine pre-filter machine, were positive for L. monocytogenes. Nineteen of the 21 (90.5%) plants had positive environmental sites. Sixty-three of the 163 (41.1%) samples from environmental sites were Listeria positive and 24 were positive for L. monocytogenes. Two-tailed student t-test analysis of the mean frequencies indicated that the level of contamination was significantly higher (p < 0.001) in 'environmental' (49.7%) as opposed to 'equipment' samples (7.0%). Our study indicates that environmental contamination with Listeria does not necessarily translate into contamination of equipment within the same plant, and that greater emphasis needs to be placed on the cleaning and sanitizing of the plant environment.

A Nationwide Survey on the Prevalence of Intestinal Parasitic Infections in the Republic of Korea, 2004

PubMed Central

Kim, Tong-Soo; Cho, Shin-Hyeong; Huh, Sun; Kong, Yoon; Sohn, Woon-Mok; Hwang, Seung-Sik; Chai, Jong-Yil; Lee, Soon-Hyung; Park, Yun-Kyu; Oh, Dae-Kyu

2009-01-01

National surveys on the prevalence of intestinal parasitic infections have been carried out every 5-7 years since 1971 in the Republic of Korea in order to establish control measures. The present nationwide survey was conducted from June to December 2004. The 10% population sampling data of Population and Housing Census by the Korean government in 2000 was used as the survey population. One sample was selected randomly from each of the 22,858 registered subjects, and a total of 20,541 people were ultimately included in this survey. Fecal examinations were performed by the cellophane thick smear and saturated brine flotation techniques. Pinworm infection was examined by cello-tape anal swab method. This survey also included a questionnaire study for a socioeconomic analysis. The total helminth egg positive rate was 3.7%, and the estimated total positive number among nationwide people was 1,780,000. The rates in urban and rural areas were 3.1% and 6.8%, respectively. As the total egg positive rate in the 6th survey in 1997 was 2.4%, the present survey showed that there was a considerable degree of increase in the prevalence rate of intestinal parasitic infections over the 7-year period following the 6th survey. The largest increases occurred in the egg positive rates of Clonorchis sinensis and heterophyids including Metagonimus yokogawai. PMID:19290090

Total and inorganic arsenic in fish samples from Norwegian waters.

PubMed

Julshamn, Kaare; Nilsen, Bente M; Frantzen, Sylvia; Valdersnes, Stig; Maage, Amund; Nedreaas, Kjell; Sloth, Jens J

2012-01-01

The contents of total arsenic and inorganic arsenic were determined in fillet samples of Northeast Artic cod, herring, mackerel, Greenland halibut, tusk, saithe and Atlantic halibut. In total, 923 individual fish samples were analysed. The fish were mostly caught in the open sea off the coast of Norway, from 40 positions. The determination of total arsenic was carried out by inductively coupled plasma mass spectrometry following microwave-assisted wet digestion. The determination of inorganic arsenic was carried out by high-performance liquid chromatography-ICP-MS following microwave-assisted dissolution of the samples. The concentrations found for total arsenic varied greatly between fish species, and ranged from 0.3 to 110 mg kg(-1) wet weight. For inorganic arsenic, the concentrations found were very low (<0.006 mg kg(-1)) in all cases. The obtained results question the assumptions made by the European Food Safety Authority (EFSA) on the inorganic arsenic level in fish used in the recent EFSA opinion on arsenic in food.

Prevalence of Campylobacter spp., Escherichia coli, and Salmonella serovars in retail chicken, turkey, pork, and beef from the Greater Washington, D.C., area.

PubMed

Zhao, C; Ge, B; De Villena, J; Sudler, R; Yeh, E; Zhao, S; White, D G; Wagner, D; Meng, J

2001-12-01

A total of 825 samples of retail raw meats (chicken, turkey, pork, and beef) were examined for the presence of Escherichia coli and Salmonella serovars, and 719 of these samples were also tested for Campylobacter spp. The samples were randomly obtained from 59 stores of four supermarket chains during 107 sampling visits in the Greater Washington, D.C., area from June 1999 to July 2000. The majority (70.7%) of chicken samples (n = 184) were contaminated with Campylobacter, and a large percentage of the stores visited (91%) had Campylobacter-contaminated chickens. Approximately 14% of the 172 turkey samples yielded Campylobacter, whereas fewer pork (1.7%) and beef (0.5%) samples were positive for this pathogen. A total of 722 Campylobacter isolates were obtained from 159 meat samples; 53.6% of these isolates were Campylobacter jejuni, 41.3% were Campylobacter coli, and 5.1% were other species. Of the 212 chicken samples, 82 (38.7%) yielded E. coli, while 19.0% of the beef samples, 16.3% of the pork samples, and 11.9% of the turkey samples were positive for E. coli. However, only 25 (3.0%) of the retail meat samples tested were positive for Salmonella. Significant differences in the bacterial contamination rates were observed for the four supermarket chains. This study revealed that retail raw meats are often contaminated with food-borne pathogens; however, there are marked differences in the prevalence of such pathogens in different meats. Raw retail meats are potential vehicles for transmitting food-borne diseases, and our findings stress the need for increased implementation of hazard analysis of critical control point (HACCP) and consumer food safety education efforts.

Prevalence of Campylobacter spp., Escherichia coli, and Salmonella Serovars in Retail Chicken, Turkey, Pork, and Beef from the Greater Washington, D.C., Area

PubMed Central

Zhao, Cuiwei; Ge, Beilei; De Villena, Juan; Sudler, Robert; Yeh, Emily; Zhao, Shaohua; White, David G.; Wagner, David; Meng, Jianghong

2001-01-01

A total of 825 samples of retail raw meats (chicken, turkey, pork, and beef) were examined for the presence of Escherichia coli and Salmonella serovars, and 719 of these samples were also tested for Campylobacter spp. The samples were randomly obtained from 59 stores of four supermarket chains during 107 sampling visits in the Greater Washington, D.C., area from June 1999 to July 2000. The majority (70.7%) of chicken samples (n = 184) were contaminated with Campylobacter, and a large percentage of the stores visited (91%) had Campylobacter-contaminated chickens. Approximately 14% of the 172 turkey samples yielded Campylobacter, whereas fewer pork (1.7%) and beef (0.5%) samples were positive for this pathogen. A total of 722 Campylobacter isolates were obtained from 159 meat samples; 53.6% of these isolates were Campylobacter jejuni, 41.3% were Campylobacter coli, and 5.1% were other species. Of the 212 chicken samples, 82 (38.7%) yielded E. coli, while 19.0% of the beef samples, 16.3% of the pork samples, and 11.9% of the turkey samples were positive for E. coli. However, only 25 (3.0%) of the retail meat samples tested were positive for Salmonella. Significant differences in the bacterial contamination rates were observed for the four supermarket chains. This study revealed that retail raw meats are often contaminated with food-borne pathogens; however, there are marked differences in the prevalence of such pathogens in different meats. Raw retail meats are potential vehicles for transmitting food-borne diseases, and our findings stress the need for increased implementation of hazard analysis of critical control point (HACCP) and consumer food safety education efforts. PMID:11722889

Perfluoroalkyl substances and lipid concentrations in plasma during pregnancy among women in the Norwegian Mother and Child Cohort Study

PubMed Central

Starling, Anne P.; Engel, Stephanie M.; Whitworth, Kristina W.; Richardson, David B.; Stuebe, Alison M.; Daniels, Julie L.; Haug, Line Småstuen; Eggesbø, Merete; Becher, Georg; Sabaredzovic, Azemira; Thomsen, Cathrine; Wilson, Ralph E.; Travlos, Gregory S.; Hoppin, Jane A.; Baird, Donna D.; Longnecker, Matthew P.

2013-01-01

Background Perfluoroalkyl substances (PFASs) are widespread and persistent environmental pollutants. Previous studies, primarily among non-pregnant individuals, suggest positive associations between PFAS levels and certain blood lipids. If there is a causal link between PFAS concentrations and elevated lipids during pregnancy, this may suggest a mechanism by which PFAS exposure leads to certain adverse pregnancy outcomes, including preeclampsia. Methods This cross-sectional analysis included 891 pregnant women enrolled in the Norwegian Mother and Child (MoBa) Cohort Study in 2003–2004. Non-fasting plasma samples were obtained at mid-pregnancy and analyzed for nineteen PFASs. Total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein cholesterol, and triglycerides were measured in plasma. Linear regression was used to quantify associations between each PFAS exposure and each lipid outcome. A multiple PFAS model was also fitted. Results Seven PFASs were quantifiable in >50% of samples. Perfluorooctane sulfonate (PFOS) concentration was associated with total cholesterol, which increased 4.2 mg/dL per interquartile shift (95% CI=0.8, 7.7) in adjusted models. Five of the seven PFASs studied were positively associated with HDL cholesterol, and all seven had elevated HDL associated with the highest quartile of exposure. Perfluoroundecanoic acid showed the strongest association with HDL: HDL increased 3.7 mg/dL per interquartile shift (95% CI=2.5, 4.9). Conclusion Plasma concentrations of PFASs were positively associated with HDL cholesterol, and PFOS was positively associated with total cholesterol in this sample of pregnant Norwegian women. While elevated HDL is not an adverse outcome per se, elevated total cholesterol associated with PFASs during pregnancy could be of concern if causal. PMID:24189199

Perfluoroalkyl substances and lipid concentrations in plasma during pregnancy among women in the Norwegian Mother and Child Cohort Study.

PubMed

Starling, Anne P; Engel, Stephanie M; Whitworth, Kristina W; Richardson, David B; Stuebe, Alison M; Daniels, Julie L; Haug, Line Småstuen; Eggesbø, Merete; Becher, Georg; Sabaredzovic, Azemira; Thomsen, Cathrine; Wilson, Ralph E; Travlos, Gregory S; Hoppin, Jane A; Baird, Donna D; Longnecker, Matthew P

2014-01-01

Perfluoroalkyl substances (PFASs) are widespread and persistent environmental pollutants. Previous studies, primarily among non-pregnant individuals, suggest positive associations between PFAS levels and certain blood lipids. If there is a causal link between PFAS concentrations and elevated lipids during pregnancy, this may suggest a mechanism by which PFAS exposure leads to certain adverse pregnancy outcomes, including preeclampsia. This cross-sectional analysis included 891 pregnant women enrolled in the Norwegian Mother and Child (MoBa) Cohort Study in 2003-2004. Non-fasting plasma samples were obtained at mid-pregnancy and analyzed for nineteen PFASs. Total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein cholesterol, and triglycerides were measured in plasma. Linear regression was used to quantify associations between each PFAS exposure and each lipid outcome. A multiple PFAS model was also fitted. Seven PFASs were quantifiable in >50% of samples. Perfluorooctane sulfonate (PFOS) concentration was associated with total cholesterol, which increased 4.2mg/dL per inter-quartile shift (95% CI=0.8, 7.7) in adjusted models. Five of the seven PFASs studied were positively associated with HDL cholesterol, and all seven had elevated HDL associated with the highest quartile of exposure. Perfluoroundecanoic acid showed the strongest association with HDL: HDL increased 3.7 mg/dL per inter-quartile shift (95% CI=2.5, 4.9). Plasma concentrations of PFASs were positively associated with HDL cholesterol, and PFOS was positively associated with total cholesterol in this sample of pregnant Norwegian women. While elevated HDL is not an adverse outcome per se, elevated total cholesterol associated with PFASs during pregnancy could be of concern if causal. © 2013.

Screening and confirmatory methods for the analysis of macrocyclic lactone mycotoxins by CE with amperometric detection.

PubMed

Arribas, Alberto Sánchez; Bermejo, Esperanza; Zapardiel, Antonio; Téllez, Helena; Rodríguez-Flores, Juana; Zougagh, Mohammed; Ríos, Angel; Chicharro, Manuel

2009-02-01

A simple analytical scheme for the screening and quantification of zearalenone and its metabolites, alpha-zearalenol and beta-zearalenol, is reported. Extracts from maize flour samples were collected by supercritical fluid extraction and afterwards, they were analyzed by CE with amperometric detection. This scheme allowed a rapid and reliable identification of contaminated flour samples according to the reference value established for zearalenone by directive 2005/38/EC (200 microg/kg). The sample screening method was carried out by CZE using 25 mM borate separation buffer at pH 9.2 and 25.0 kV as separation voltage, monitoring the amperometric signal at +700 mV with a carbon paste electrode. In this way, total amount of mycotoxins was determined and samples were processed in 4 min with a detection limit of 12 microg/L, enough to discriminate between positive (more than 200 microg/L total mycotoxins) and negative samples (less than 200 microg/L total mycotoxins). Positive samples were then subjected to CZE separation and quantification of each analyte was done with 50 mM borate running buffer modified with 30% methanol at pH 9.7 and 17.5 kV as separation voltage. Under these conditions, separation was achieved in 15 min with detection limits from 20 to 35 microg/L for each analyte.

Crypto-Giardia antigen rapid test versus conventional modified Ziehl-Neelsen acid fast staining method for diagnosis of cryptosporidiosis.

PubMed

Zaglool, Dina Abdulla Muhammad; Mohamed, Amr; Khodari, Yousif Abdul Wahid; Farooq, Mian Usman

2013-03-01

To evaluate the validity of Crypto-Giardia antigen rapid test (CA-RT) in comparison with the conventional modified Ziehl-Neelsen acid fast (MZN-AF) staining method for the diagnosis of cryptosporidiosis. Fifteen preserved stool samples from previously confirmed infections were used as positive controls and 40 stool samples from healthy people were used as negative control. A total of 85 stool samples were collected from suspected patients with cryptosporidiosis over 6 months during the period from January till June, 2011. The study was conducted in the department of parasitology, central laboratory, Alnoor Specialist Hospital, Makkah, Saudi Arabia. All samples were subjected to CA-RT and conventional MZN-AF staining method. Validation parameters including sensitivity (SN), specificity (SP), accuracy index (AI), positive predictive value (PPV), and negative predictive value (NPV) were evaluated for both tests. Out of 15 positive controls, CA-RT detected 13 (86.7%) while MZN-AF detected 11(73.3%) positive cases. However, CA-RT detected no positive case in 40 normal controls but MZN-AF detected 2(5%) as positive cases. Based on the results, the SN, SP, AI, PPV and NPV were high in CA-RT than MZN-AF staining method, ie., 86.7%vs. 73.3%, 100%vs. 95%, 96.4%vs. 89.1%, 100%vs. 84.6% and 95.2%vs. 90.5%, respectively. Out of a total of 85 suspected specimens, CA-RT detected 7(8.2%) but MZN-AF detected 6(7.1%) cases as positive. CA-RT immunoassay is more valid and reliable than MZN-AF staining method. Copyright © 2013 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Survey of feline leukemia virus and feline coronaviruses in captive neotropical wild felids from Southern Brazil.

PubMed

Guimaraes, Ana M S; Brandão, Paulo E; de Moraes, Wanderlei; Cubas, Zalmir S; Santos, Leonilda C; Villarreal, Laura Y B; Robes, Rogério R; Coelho, Fabiana M; Resende, Mauricio; Santos, Renata C F; Oliveira, Rosangela C; Yamaguti, Mauricio; Marques, Lucas M; Neto, Renata L; Buzinhani, Melissa; Marques, Regina; Messick, Joanne B; Biondo, Alexander W; Timenetsky, Jorge

2009-06-01

A total of 57 captive neotropical felids (one Leopardus geoffroyi, 14 Leopardus pardalis, 17 Leopardus wiedii, 22 Leopardus tigrinus, and three Puma yagouaroundi) from the Itaipu Binacional Wildlife Research Center (Refúgio Bela Vista, Southern Brazil) were anesthetized for blood collection. Feces samples were available for 44 animals, including one L. geoffroyi, eight L. pardalis, 14 L. wiedii, 20 L. tigrinus, and one P. yagouaroundi. Total DNA and RNA were extracted from blood and feces, respectively, using commercial kits. Blood DNA samples were evaluated by polymerase chain reaction (PCR) for feline leukemia virus (FeLV) proviral DNA, whereas reverse transcriptase-PCR was run on fecal samples for detection of coronavirus RNA. None of the samples were positive for coronaviruses. A male L. pardalis and a female L. tigrinus were positive for FeLV proviral DNA, and identities of PCR products were confirmed by sequencing. This is the first evidence of FeLV proviral DNA in these species in Southern Brazil.

Opportunistic Parasites among Immunosuppressed Children in Minia District, Egypt

PubMed Central

Ahmad, Azza K.; Ali, Basma A.; Moslam, Fadia A.

2012-01-01

A total of 450 stool samples were collected from inpatient and outpatient clinics of Pediatric Department, Minia University Hospital, Minia District, Egypt. Two groups of patients were studied, including 200 immunosuppressed and 250 immunocompetent children. Stool samples were subjected to wet saline and iodine mounts. A concentration technique (formol-ether sedimentation method) was carried out for stool samples diagnosed negative by wet saline and iodine mounts. Samples were stained by 2 different methods; acid fast stain (modified Ziehl-Neelsen stain) and Giemsa stain. Total 188 cases (94%) were diagnosed positive for parasitic infections among immunosuppressed children, whereas 150 cases (60%) were positive in immunocompetent children (P<0.0001). The most common protozoan infection in immunosuppressed group was Cryptosporidium parvum (60.2%), followed by Blastocystis hominis (12.1%), Isospora belli (9.7%), and Cyclospora caytenensis (7.8%). On the other hand, Entamoeba histolytica (24.6%) and Giardia lamblia (17.6%) were more common than other protozoans in immunocompetent children. PMID:22451735

Assessing Positivity and Circulating Levels of NS1 in Samples from a 2012 Dengue Outbreak in Rio de Janeiro, Brazil

PubMed Central

Allonso, Diego; Meneses, Marcelo D. F.; Fernandes, Carlos A.; Ferreira, Davis F.; Mohana-Borges, Ronaldo

2014-01-01

Dengue virus (DENV) represents a major threat to public health worldwide. Early DENV diagnosis should not only detect the infection but also identify patients with a higher likelihood to develop severe cases. Previous studies have suggested the potential for NS1 to serve as a viral marker for dengue severity. However, further studies using different sera panels are required to confirm this hypothesis. In this context, we developed a lab-based ELISA to detect and quantitate NS1 protein from the four DENV serotypes and from primary and secondary cases. This approach was used to calculate the circulating NS1 concentration in positive samples. We also tested the NS1 positivity of DENV-positive samples according to the Platelia Dengue NS1 Ag assay. A total of 128 samples were positive for DENV infection and were classified according to the WHO guidelines. The overall NS1 positivity was 68% according to the Platelia assay, whereas all samples were NS1-positive when analyzed with our lab-based ELISA. Fifty-four samples were positive by PCR, revealing a co-circulation of DENV1 and DENV4, and the NS1 positivity for DENV4 samples was lower than that for DENV1. The circulating NS1 concentration ranged from 7 to 284 ng/mL. Our results support previous data indicating the low efficiency of the Platelia assay to detect DENV4 infection. Moreover, this work is the first to analyze NS1 antigenemia using retrospective samples from a Brazilian outbreak. PMID:25412084

A strategical re-thinking on National Blood Donor Pool: Anti-HBc positivity related re-entry mechanisms.

PubMed

Yilmaz, Soner; Unlu, Aytekin; Cetinkaya, Riza Aytac; Yapar, Mehmet; Avci, Ismail Yasar; Yilmaz, Sebahattin; Eyigun, Can Polat

2016-04-01

Screening of blood donations for antibodies against hepatitis B core antigen (anti-HBc) is used to prevent transfusion transmitted hepatitis B virus (HBV) infection. In this study, we studied the magnitude of blood donor gain by using a re-entry mechanism in our Blood Bank of Gulhane Military Academy of Medicine. Between January and May 2013, 5148 voluntary blood donors were screened by ELISA method for HBsAg, anti-HBc total and other screening markers, prospectively. Samples with repeated reactivity for the presence of anti-HBc were further tested with four supplemental assays. We detected 515 (10%) anti-HBc positive and 4612 (90%) anti-HBc negative cases in 5127 HBsAg negative serum samples. A total of 461 (89.5%) blood units were reactive for at least one additional serologic parameter and 54 were (10.5%) negative. Isolated anti-HBc positivity rate was 1.3% (69/5127). In the isolated anti-HBc positive samples, 54 were also anti-HBe and HBeAg negative. HBV DNA was not detected in any of the samples. Applying the EDQM criteria would decrease our blood donor loss from 10% to 5.4%. As alternative re-entry mechanisms have already been presented in the literature, institution of a new policy is needed to enhance the limited blood donor pool in our system. Copyright © 2016. Published by Elsevier Ltd.

Exploring the Legionella pneumophila positivity rate in hotel water samples from Antalya, Turkey.

PubMed

Sepin Özen, Nevgün; Tuğlu Ataman, Şenay; Emek, Mestan

2017-05-01

The genus Legionella is a fastidious Gram-negative bacteria widely distributed in natural waters and man made water supply systems. Legionella pneumophila is the aetiological agent of approximately 90% of reported Legionellosis cases, and serogroup 1 is the most frequent cause of infections. Legionnaires' disease is often associated with travel and continues to be a public health concern at present. The correct water management quality practices and rapid methods for analyzing Legionella species in environmental water is a key point for the prevention of Legionnaires' disease outbreaks. This study aimed to evaluate the positivity rates and serotyping of Legionella species from water samples in the region of Antalya, Turkey, which is an important tourism center. During January-December 2010, a total of 1403 samples of water that were collected from various hotels (n = 56) located in Antalya were investigated for Legionella pneumophila. All samples were screened for L. pneumophila by culture method according to "ISO 11731-2" criteria. The culture positive Legionella strains were serologically identified by latex agglutination test. A total of 142 Legionella pneumophila isolates were recovered from 21 (37.5%) of 56 hotels. The total frequency of L. pneumophila isolation from water samples was found as 10.1%. Serological typing of 142 Legionella isolates by latex agglutination test revealed that strains belonging to L. pneumophila serogroups 2-14 predominated in the examined samples (85%), while strains of L. pneumophila serogroup 1 were less numerous (15%). According to our knowledge, our study with the greatest number of water samples from Turkey demonstrates that L. pneumophila serogroups 2-14 is the most common isolate. Rapid isolation of L. pneumophila from environmental water samples is essential for the investigation of travel related outbreaks and the possible resources. Further studies are needed to have epidemiological data and to determine the types of L. pneumophila isolates from Turkey.

Detection of Mycoplasma pneumoniae and Legionella pneumophila in Patients Having Community-Acquired Pneumonia: A Multicentric Study from New Delhi, India.

PubMed

Chaudhry, Rama; Valavane, Arvind; Sreenath, K; Choudhary, Mamta; Sagar, Tanu; Shende, Trupti; Varma-Basil, Mandira; Mohanty, Srujana; Kabra, S K; Dey, A B; Thakur, Bhaskar

2017-12-01

Atypical pathogens including Mycoplasma pneumoniae and Legionella pneumophila are increasingly recognized as important causes of community-acquired pneumonia (CAP). Mycoplasma pneumoniae accounts for 20-40% of all CAP and L. pneumophila is responsible for 3-15% of cases. The paucity of data from India in this regard prompted us to conduct this prospective multicentric analysis to detect the prevalence of M. pneumoniae and L. pneumophila in our geographical region. A total of 453 patients with symptoms of pneumonia and 90 controls with no history of lower respiratory tract infections were included in the study. A duplex polymerase chain reaction (PCR) targeting 543 bp region of P1 adhesin gene of M. pneumoniae and 375 bp region of macrophage infectivity potentiator (mip) gene of L. pneumophila was standardized for simultaneous detection of these atypical pathogens. Respiratory secretions, blood, and urine samples were collected from each patient and control and were subjected to duplex PCR, culture and serology for M. pneumoniae and L. pneumophila . Urine samples were subjected for detecting L. pneumophila antigen. Among the 453 patients investigated for M. pneumoniae , 52 (11.4%) were positive for IgM antibodies, 17 were positive by culture, and seven tested positive by PCR ( P1 gene). Similarly for L. pneumophila , 50 cases (11%) were serologically positive for IgM antibodies, one was positive by PCR ( mip gene) and urine antigen detection. A total of eight samples were positive by duplex PCR for M. pneumoniae P1 gene ( N = 7) and L. pneumophila mip gene ( N = 1). Of the 90 controls, two samples (2.2%) showed IgM positivity, and 15 (16.7%) showed IgG positivity for M. pneumoniae . For L. pneumophila , three samples (3.3%) tested positive for IgM, and 12 (13.3%) tested positive for IgG antibodies. The study findings indicate the presence of M. pneumoniae and L. pneumophila in our geographical region, and a combination of laboratory approaches including PCR, culture, and serology is required for effective detection of these agents.

Absence of Plasmodium inui and Plasmodium cynomolgi, but detection of Plasmodium knowlesi and Plasmodium vivax infections in asymptomatic humans in the Betong division of Sarawak, Malaysian Borneo.

PubMed

Siner, Angela; Liew, Sze-Tze; Kadir, Khamisah Abdul; Mohamad, Dayang Shuaisah Awang; Thomas, Felicia Kavita; Zulkarnaen, Mohammad; Singh, Balbir

2017-10-17

Plasmodium knowlesi, a simian malaria parasite, has become the main cause of malaria in Sarawak, Malaysian Borneo. Epidemiological data on malaria for Sarawak has been derived solely from hospitalized patients, and more accurate epidemiological data on malaria is necessary. Therefore, a longitudinal study of communities affected by knowlesi malaria was undertaken. A total of 3002 blood samples on filter paper were collected from 555 inhabitants of 8 longhouses with recently reported knowlesi malaria cases in the Betong Division of Sarawak, Malaysian Borneo. Each longhouse was visited bimonthly for a total of 10 times during a 21-month study period (Jan 2014-Oct 2015). DNA extracted from blood spots were examined by a nested PCR assay for Plasmodium and positive samples were then examined by nested PCR assays for Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale, Plasmodium knowlesi, Plasmodium cynomolgi and Plasmodium inui. Blood films of samples positive by PCR were also examined by microscopy. Genus-specific PCR assay detected Plasmodium DNA in 9 out of 3002 samples. Species-specific PCR identified 7 P. knowlesi and one P. vivax. Malaria parasites were observed in 5 thick blood films of the PCR positive samples. No parasites were observed in blood films from one knowlesi-, one vivax- and the genus-positive samples. Only one of 7 P. knowlesi-infected individual was febrile and had sought medical treatment at Betong Hospital the day after sampling. The 6 knowlesi-, one vivax- and one Plasmodium-infected individuals were afebrile and did not seek any medical treatment. Asymptomatic human P. knowlesi and P. vivax malaria infections, but not P. cynomolgi and P. inui infections, are occurring within communities affected with malaria.

Detection, identification, and differentiation of sheep pox virus and goat pox virus from clinical cases in Giza Governorate, Egypt

PubMed Central

Mahmoud, M. A.; Khafagi, M. H.

2016-01-01

Aim: To isolate, identify, and differentiate Capripoxviruses (CaPV) (sheep pox virus and goat pox virus) infections by egg inoculation, transmission electron microscopy (TEM), and 30 kDa RNA polymerase subunit gene-based polymerase chain reaction (PCR) (RPO30) in clinically affected animals in Hawamdia township of Giza Governorate, Egypt. Materials and Methods: A total of 37 scab samples were collected from clinically suspected field cases of sheep pox and goat pox. These samples were collected during (2014-2015) during different outbreaks of sheep pox and goat pox from Hawamdia township of Giza Governorate, Egypt. The samples were subjected to egg inoculation, TEM, and (RPO30) gene-based PCR. By using the egg inoculation: Previously prepared 37 scab samples (n=23 sheep and n=14 goats) were inoculated on the chorioallantoic membrane of specific pathogen free (SPF) embryonated chicken eggs (12 days old age). In the presence of the suitable percentage of humidity and candling, the inoculated eggs were incubated at 37°C. By using the TEM: Samples showed positive pock lesions on the chorioallantoic membranes, were fixed in glutaraldehyde, then processed and sectioned for TEM. Using the (RPO30) gene-based PCR assay, 30 of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened. Results: Using the egg inoculation, a characteristic pock lesions for poxviruses were seen in 30/37 (n=19 sheep and n=11 goats) (81.08%). Using the TEM, examination of the positive samples after egg inoculation revealed positive result in 23/30 (n=15 sheep and n=8 goats) (76.66%). The positive results represented by the presence of negatively stained oval-shape virus particles. Using the (RPO30) gene-based PCR assay, out of 30 total of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened, 27 (90%) samples (n=17 sheep and n=10 goats) were positive. The given band sizes of sheep and goats were 172 and 152 bp, respectively. Conclusion: PCR assay depended on RPO30 gene can be used lonely for the detection, identification, and differentiation of CaPVs. RPO30 gene-based PCR assay in combination with gene sequencing helps in molecular epidemiological studies of CaPV infection. PMID:28096619

Detection, identification, and differentiation of sheep pox virus and goat pox virus from clinical cases in Giza Governorate, Egypt.

PubMed

Mahmoud, M A; Khafagi, M H

2016-12-01

To isolate, identify, and differentiate Capripoxviruses (CaPV) (sheep pox virus and goat pox virus) infections by egg inoculation, transmission electron microscopy (TEM), and 30 kDa RNA polymerase subunit gene-based polymerase chain reaction (PCR) (RPO30) in clinically affected animals in Hawamdia township of Giza Governorate, Egypt. A total of 37 scab samples were collected from clinically suspected field cases of sheep pox and goat pox. These samples were collected during (2014-2015) during different outbreaks of sheep pox and goat pox from Hawamdia township of Giza Governorate, Egypt. The samples were subjected to egg inoculation, TEM, and (RPO30) gene-based PCR. By using the egg inoculation: Previously prepared 37 scab samples (n=23 sheep and n=14 goats) were inoculated on the chorioallantoic membrane of specific pathogen free (SPF) embryonated chicken eggs (12 days old age). In the presence of the suitable percentage of humidity and candling, the inoculated eggs were incubated at 37°C. By using the TEM: Samples showed positive pock lesions on the chorioallantoic membranes, were fixed in glutaraldehyde, then processed and sectioned for TEM. Using the (RPO30) gene-based PCR assay, 30 of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened. Using the egg inoculation, a characteristic pock lesions for poxviruses were seen in 30/37 (n=19 sheep and n=11 goats) (81.08%). Using the TEM, examination of the positive samples after egg inoculation revealed positive result in 23/30 (n=15 sheep and n=8 goats) (76.66%). The positive results represented by the presence of negatively stained oval-shape virus particles. Using the (RPO30) gene-based PCR assay, out of 30 total of positive samples after egg inoculation (n=19 sheep and n=11 goats) were screened, 27 (90%) samples (n=17 sheep and n=10 goats) were positive. The given band sizes of sheep and goats were 172 and 152 bp, respectively. PCR assay depended on RPO30 gene can be used lonely for the detection, identification, and differentiation of CaPVs. RPO30 gene-based PCR assay in combination with gene sequencing helps in molecular epidemiological studies of CaPV infection.

Molecular and serological study of rickettsial infection in humans, and in wild and farm animals, in the province of Burgos, Spain.

PubMed

Lledó, Lourdes; Domínguez-Peñafiel, Gerardo; Giménez-Pardo, Consuelo; Gegúndez, Isabel; González, Rosario; Saz, José Vicente

2014-06-01

Limited information is available on the presence of rickettsial infection in humans and animal reservoirs in Spain. Exposure to spotted fever group rickettsia in healthy humans and in farm and wild animals in the Province of Burgos, Spain, was examined by serological methods. Rickettsial DNA was also sought by PCR in animal samples. Of 102 human serum samples examined by indirect immunofluorescence assays (IFA), 5.88% were positive for antibodies against Rickettsia conorii (titers 1/128-1/512). Significant differences were detected in human seroprevalence with respect to age. In further IFAs, 102 out of 375 (27.2%) serum samples from the wild animals reacted with R. conorii antigens (titers 1/64-1/1024); 32 out of 281 (11.38%) samples from farm animals were also positive for R. conorii (titers 1/64-1/2048). The prevalence detected among total wild animals was significantly higher than among total farm animals. No rickettsial DNA was found by PCR in any farm or wild animal sample.

Serological evidence of hepatitis E virus infection in dromedary camels in Ethiopia.

PubMed

Li, Tian-Cheng; Yoshizaki, Sayaka; Zhou, Xianfeng; Sentsui, Hiroshi; Shirato, Kazuya; Matsuyama, Shutoku; Melaku, Simenew Keskes; Bazartseren, Boldbaatar; Takeda, Naokazu; Wakita, Takaji

2017-08-01

The genome of dromedary camel hepatitis E virus (DcHEV) has been detected in stool and serum samples from dromedary camels, but the sero-epidemiological information of DcHEV infection remains unclear. A total of 246 serum samples collected from dromedary camels (Camelus dromedarius) in Ethiopia, and 40 serum samples from Bactrian camels (Camelus ferus) in Mongolia were examined for the detection of anti-DcHEV IgG antibody by a newly developed enzyme-linked immunosorbent assay (ELISA) by using DcHEV-like particles (DcHEV-LPs) as the antigen. The results revealed that 55 of the 246 (22.4%) dromedary camels were positive for anti-DcHEV IgG, whereas all 40 samples from the Bactrian camels were negative for DcHEV IgG antibody. A total of 98 serum samples from dromedary camels, including 25 anti-DcHEV-IgG positive samples, were used for the detection of DcHEV RNA by reverse transcription-polymerase chain reaction (RT-PCR), however, no positive samples were identified. These results suggested that the DcHEV infection occurred in the dromedary camels in Ethiopia. Further studies are required to determine whether Bactrian camels are susceptible to DcHEV infection. In addition, not only DcHEV-LPs, but also virus-like particles (VLPs) delivered from G1, G3 and G5 HEV are likely applicable for the detection of the anti-DcHEV IgG antibody. Copyright © 2017 Elsevier B.V. All rights reserved.

Bacteriological quality of bottled drinking water versus municipal tap water in Dharan municipality, Nepal.

PubMed

Pant, Narayan Dutt; Poudyal, Nimesh; Bhattacharya, Shyamal Kumar

2016-06-07

Water-related diseases are of great concern in developing countries like Nepal. Every year, there are countless morbidity and mortality due to the consumption of unsafe drinking water. Recently, there have been increased uses of bottled drinking water in an assumption that the bottled water is safer than the tap water and its use will help to protect from water-related diseases. So, the main objective of this study was to analyze the bacteriological quality of bottled drinking water and that of municipal tap water. A total of 100 samples (76 tap water and 24 bottled water) were analyzed for bacteriological quality and pH. The methods used were spread plate method for total plate count (TPC) and membrane filter method for total coliform count (TCC), fecal coliform count (FCC), and fecal streptococcal count (FSC). pH meter was used for measuring pH. One hundred percent of the tap water samples and 87.5 % of the bottled water samples were found to be contaminated with heterotrophic bacteria. Of the tap water samples, 55.3 % were positive for total coliforms, compared with 25 % of the bottled water. No bottled water samples were positive for fecal coliforms and fecal streptococci, in contrast to 21.1 % and 14.5 % of the tap water samples being contaminated with fecal coliforms and fecal streptococci, respectively. One hundred percent of the tap water samples and 54.2 % of the bottled water samples had pH in the acceptable range. All of the municipal tap water samples and most of the bottled drinking water samples distributed in Dharan municipality were found to be contaminated with one or more than one type of indicator organisms. On the basis of our findings, we may conclude that comparatively, the bottled drinking water may have been safer (than tap water) to drink.

Synthetic food colours in saffron solutions, saffron rice and saffron chicken from restaurants in Tehran, Iran.

PubMed

Moradi-Khatoonabadi, Zhila; Amirpour, Mansooreh; AkbariAzam, Maryam

2015-01-01

Saffron solutions, saffron rice and saffron chicken samples were considered for synthetic colours as additives, which are forbidden according to Iranian national standards. Samples were taken from restaurants of three locations and analysed by high-performance liquid chromatography. Of the total 573 samples, 52% were positive for at least one colour. The most prevalent colours were Tartrazine, Quinoline Yellow and Sunset Yellow, with 44%, 9.1% and 8.4% of the samples testing positive for these colours, respectively. Carmoisine and Ponceau were both detected only in 0.5% of the positive samples and found only in saffron solution. In conclusion, synthetic food colours, especially Tartrazine should be regarded as a potential risk in saffron and its related food. Therefore, new attempts for food safety and quality should be undertaken to eliminate the use of these colours in restaurants.

Reverse line blot hybridisation screening of Pseudallescheria/Scedosporium species in patients with cystic fibrosis.

PubMed

Lu, Q; van den Ende, A H G Gerrits; de Hoog, G S; Li, R; Accoceberry, I; Durand-Joly, I; Bouchara, J-P; Hernandez, F; Delhaes, L

2011-10-01

The PCR-RLB (reverse line blot hybridisation) was applied as a molecular technique for the detection of members of Pseudallescheria and Scedosporium from sputum of patients with cystic fibrosis (CF). Fifty-nine sputum samples were collected from 52 CF patients, which were analysed by culture and PCR-RLB. Conventional and semi-selective culture yielded five positive samples, but the PCR-RLB hybridisation assay permitted the detection of members of Pseudallescheria/Scedosporium in 32 out of 52 patients (61.5%). In total, PCR-RLB yielded 47 positives. Pseudallescheria apiosperma was detected in 20 samples, while Pseudallescheria boydii and Pseudallescheria aurantiacum were detected in 17 and eight samples, respectively. Six samples gave a positive reaction with two distinct species-specific probes and one sample with three probes. In conclusion, the PCR-RLB assay described in this study allows the detection of Scedosporium spp. in CF sputum samples and the identification of Pseudallescheria apiosperma, P. boydii, S. aurantiacum, Scedosporium prolificans and Pseudallescheria minutispora. © 2011 Blackwell Verlag GmbH.

17 CFR Appendix B to Part 420 - Sample Large Position Report

Code of Federal Regulations, 2014 CFR

2014-04-01

... Memorandum 1 $ Memorandum 2: Report the gross par amount of fails to deliver. Included in the calculation of... millions at par value as of trade date] Security Being Reported Date For Which Information is Being... Principal Components of the Specific Security $________ Total Net Trading Position $ 2. Gross Financing...

Comparison of Clark's presence-absence test and the membrane filter method for coliform detection in potable water samples.

PubMed Central

Pipes, W O; Minnigh, H A; Moyer, B; Troy, M A

1986-01-01

A total of 2,601 water samples from six different water systems were tested for coliform bacteria by Clark's presence-absence (P-A) test and by the membrane filter (MF) method. There was no significant difference in the fraction of samples positive for coliform bacteria for any of the systems tested. It was concluded that the two tests are equivalent for monitoring purposes. However, 152 samples were positive for coliform bacteria by the MF method but negative by the P-A test, and 132 samples were positive by the P-A test but negative by the MF method. Many of these differences for individual samples can be explained by random dispersion of bacteria in subsamples when the coliform density is low. However, 15 samples had MF counts greater than 3 and gave negative P-A results. The only apparent explanation for most of these results is that coliform bacteria were present in the P-A test bottles but did not produce acid and gas. Two other studies have reported more samples positive by Clark's P-A test than by the MF method. PMID:3532953

Dynamic X-ray diffraction sampling for protein crystal positioning

DOE Office of Scientific and Technical Information (OSTI.GOV)

Scarborough, Nicole M.; Godaliyadda, G. M. Dilshan P.; Ye, Dong Hye

A sparse supervised learning approach for dynamic sampling (SLADS) is described for dose reduction in diffraction-based protein crystal positioning. Crystal centering is typically a prerequisite for macromolecular diffraction at synchrotron facilities, with X-ray diffraction mapping growing in popularity as a mechanism for localization. In X-ray raster scanning, diffraction is used to identify the crystal positions based on the detection of Bragg-like peaks in the scattering patterns; however, this additional X-ray exposure may result in detectable damage to the crystal prior to data collection. Dynamic sampling, in which preceding measurements inform the next most information-rich location to probe for image reconstruction,more » significantly reduced the X-ray dose experienced by protein crystals during positioning by diffraction raster scanning. The SLADS algorithm implemented herein is designed for single-pixel measurements and can select a new location to measure. In each step of SLADS, the algorithm selects the pixel, which, when measured, maximizes the expected reduction in distortion given previous measurements. Ground-truth diffraction data were obtained for a 5 µm-diameter beam and SLADS reconstructed the image sampling 31% of the total volume and only 9% of the interior of the crystal greatly reducing the X-ray dosage on the crystal. Furthermore, by usingin situtwo-photon-excited fluorescence microscopy measurements as a surrogate for diffraction imaging with a 1 µm-diameter beam, the SLADS algorithm enabled image reconstruction from a 7% sampling of the total volume and 12% sampling of the interior of the crystal. When implemented into the beamline at Argonne National Laboratory, without ground-truth images, an acceptable reconstruction was obtained with 3% of the image sampled and approximately 5% of the crystal. The incorporation of SLADS into X-ray diffraction acquisitions has the potential to significantly minimize the impact of X-ray exposure on the crystal by limiting the dose and area exposed for image reconstruction and crystal positioning using data collection hardware present in most macromolecular crystallography end-stations.« less

Dynamic X-ray diffraction sampling for protein crystal positioning

DOE PAGES

Scarborough, Nicole M.; Godaliyadda, G. M. Dilshan P.; Ye, Dong Hye; ...

2017-01-01

A sparse supervised learning approach for dynamic sampling (SLADS) is described for dose reduction in diffraction-based protein crystal positioning. Crystal centering is typically a prerequisite for macromolecular diffraction at synchrotron facilities, with X-ray diffraction mapping growing in popularity as a mechanism for localization. In X-ray raster scanning, diffraction is used to identify the crystal positions based on the detection of Bragg-like peaks in the scattering patterns; however, this additional X-ray exposure may result in detectable damage to the crystal prior to data collection. Dynamic sampling, in which preceding measurements inform the next most information-rich location to probe for image reconstruction,more » significantly reduced the X-ray dose experienced by protein crystals during positioning by diffraction raster scanning. The SLADS algorithm implemented herein is designed for single-pixel measurements and can select a new location to measure. In each step of SLADS, the algorithm selects the pixel, which, when measured, maximizes the expected reduction in distortion given previous measurements. Ground-truth diffraction data were obtained for a 5 µm-diameter beam and SLADS reconstructed the image sampling 31% of the total volume and only 9% of the interior of the crystal greatly reducing the X-ray dosage on the crystal. Furthermore, by usingin situtwo-photon-excited fluorescence microscopy measurements as a surrogate for diffraction imaging with a 1 µm-diameter beam, the SLADS algorithm enabled image reconstruction from a 7% sampling of the total volume and 12% sampling of the interior of the crystal. When implemented into the beamline at Argonne National Laboratory, without ground-truth images, an acceptable reconstruction was obtained with 3% of the image sampled and approximately 5% of the crystal. The incorporation of SLADS into X-ray diffraction acquisitions has the potential to significantly minimize the impact of X-ray exposure on the crystal by limiting the dose and area exposed for image reconstruction and crystal positioning using data collection hardware present in most macromolecular crystallography end-stations.« less

Dynamic X-ray diffraction sampling for protein crystal positioning

PubMed Central

Scarborough, Nicole M.; Godaliyadda, G. M. Dilshan P.; Ye, Dong Hye; Kissick, David J.; Zhang, Shijie; Newman, Justin A.; Sheedlo, Michael J.; Chowdhury, Azhad U.; Fischetti, Robert F.; Das, Chittaranjan; Buzzard, Gregery T.; Bouman, Charles A.; Simpson, Garth J.

2017-01-01

A sparse supervised learning approach for dynamic sampling (SLADS) is described for dose reduction in diffraction-based protein crystal positioning. Crystal centering is typically a prerequisite for macromolecular diffraction at synchrotron facilities, with X-ray diffraction mapping growing in popularity as a mechanism for localization. In X-ray raster scanning, diffraction is used to identify the crystal positions based on the detection of Bragg-like peaks in the scattering patterns; however, this additional X-ray exposure may result in detectable damage to the crystal prior to data collection. Dynamic sampling, in which preceding measurements inform the next most information-rich location to probe for image reconstruction, significantly reduced the X-ray dose experienced by protein crystals during positioning by diffraction raster scanning. The SLADS algorithm implemented herein is designed for single-pixel measurements and can select a new location to measure. In each step of SLADS, the algorithm selects the pixel, which, when measured, maximizes the expected reduction in distortion given previous measurements. Ground-truth diffraction data were obtained for a 5 µm-diameter beam and SLADS reconstructed the image sampling 31% of the total volume and only 9% of the interior of the crystal greatly reducing the X-ray dosage on the crystal. Using in situ two-photon-excited fluorescence microscopy measurements as a surrogate for diffraction imaging with a 1 µm-diameter beam, the SLADS algorithm enabled image reconstruction from a 7% sampling of the total volume and 12% sampling of the interior of the crystal. When implemented into the beamline at Argonne National Laboratory, without ground-truth images, an acceptable reconstruction was obtained with 3% of the image sampled and approximately 5% of the crystal. The incorporation of SLADS into X-ray diffraction acquisitions has the potential to significantly minimize the impact of X-ray exposure on the crystal by limiting the dose and area exposed for image reconstruction and crystal positioning using data collection hardware present in most macromolecular crystallography end-stations. PMID:28009558

Dynamic X-ray diffraction sampling for protein crystal positioning.

PubMed

Scarborough, Nicole M; Godaliyadda, G M Dilshan P; Ye, Dong Hye; Kissick, David J; Zhang, Shijie; Newman, Justin A; Sheedlo, Michael J; Chowdhury, Azhad U; Fischetti, Robert F; Das, Chittaranjan; Buzzard, Gregery T; Bouman, Charles A; Simpson, Garth J

2017-01-01

A sparse supervised learning approach for dynamic sampling (SLADS) is described for dose reduction in diffraction-based protein crystal positioning. Crystal centering is typically a prerequisite for macromolecular diffraction at synchrotron facilities, with X-ray diffraction mapping growing in popularity as a mechanism for localization. In X-ray raster scanning, diffraction is used to identify the crystal positions based on the detection of Bragg-like peaks in the scattering patterns; however, this additional X-ray exposure may result in detectable damage to the crystal prior to data collection. Dynamic sampling, in which preceding measurements inform the next most information-rich location to probe for image reconstruction, significantly reduced the X-ray dose experienced by protein crystals during positioning by diffraction raster scanning. The SLADS algorithm implemented herein is designed for single-pixel measurements and can select a new location to measure. In each step of SLADS, the algorithm selects the pixel, which, when measured, maximizes the expected reduction in distortion given previous measurements. Ground-truth diffraction data were obtained for a 5 µm-diameter beam and SLADS reconstructed the image sampling 31% of the total volume and only 9% of the interior of the crystal greatly reducing the X-ray dosage on the crystal. Using in situ two-photon-excited fluorescence microscopy measurements as a surrogate for diffraction imaging with a 1 µm-diameter beam, the SLADS algorithm enabled image reconstruction from a 7% sampling of the total volume and 12% sampling of the interior of the crystal. When implemented into the beamline at Argonne National Laboratory, without ground-truth images, an acceptable reconstruction was obtained with 3% of the image sampled and approximately 5% of the crystal. The incorporation of SLADS into X-ray diffraction acquisitions has the potential to significantly minimize the impact of X-ray exposure on the crystal by limiting the dose and area exposed for image reconstruction and crystal positioning using data collection hardware present in most macromolecular crystallography end-stations.

Romer Labs RapidChek®Listeria monocytogenes Test System for the Detection of L. monocytogenes on Selected Foods and Environmental Surfaces.

PubMed

Juck, Gregory; Gonzalez, Verapaz; Allen, Ann-Christine Olsson; Sutzko, Meredith; Seward, Kody; Muldoon, Mark T

2018-04-27

The Romer Labs RapidChek ® Listeria monocytogenes test system (Performance Tested Method ℠ 011805) was validated against the U.S. Department of Agriculture-Food Safety and Inspection Service Microbiology Laboratory Guidebook (USDA-FSIS/MLG), U.S. Food and Drug Association Bacteriological Analytical Manual (FDA/BAM), and AOAC Official Methods of Analysis ℠ (AOAC/OMA) cultural reference methods for the detection of L. monocytogenes on selected foods including hot dogs, frozen cooked breaded chicken, frozen cooked shrimp, cured ham, and ice cream, and environmental surfaces including stainless steel and plastic in an unpaired study design. The RapidChek method uses a proprietary enrichment media system, a 44-48 h enrichment at 30 ± 1°C, and detects L. monocytogenes on an immunochromatographic lateral flow device within 10 min. Different L. monocytogenes strains were used to spike each of the matrixes. Samples were confirmed based on the reference method confirmations and an alternate confirmation method. A total of 140 low-level spiked samples were tested by the RapidChek method after enrichment for 44-48 h in parallel with the cultural reference method. There were 88 RapidChek presumptive positives. One of the presumptive positives was not confirmed culturally. Additionally, one of the culturally confirmed samples did not exhibit a presumptive positive. No difference between the alternate confirmation method and reference confirmation method was observed. The respective cultural reference methods (USDA-FSIS/MLG, FDA/BAM, and AOAC/OMA) produced a total of 63 confirmed positive results. Nonspiked samples from all foods were reported as negative for L. monocytogenes by all methods. Probability of detection analysis demonstrated no significant differences in the number of positive samples detected by the RapidChek method and the respective cultural reference method.

Identification of key genes in Gram-positive and Gram-negative sepsis using stochastic perturbation

PubMed Central

Li, Zhenliang; Zhang, Ying; Liu, Yaling; Liu, Yanchun; Li, Youyi

2017-01-01

Sepsis is an inflammatory response to pathogens (such as Gram-positive and Gram-negative bacteria), which has high morbidity and mortality in critically ill patients. The present study aimed to identify the key genes in Gram-positive and Gram-negative sepsis. GSE6535 was downloaded from Gene Expression Omnibus, containing 17 control samples, 18 Gram-positive samples and 25 Gram-negative samples. Subsequently, the limma package in R was used to screen the differentially expressed genes (DEGs). Hierarchical clustering was conducted for the specific DEGs in Gram-negative and Gram-negative samples using cluster software and the TreeView software. To analyze the correlation of samples at the gene level, a similarity network was constructed using Cytoscape software. Functional and pathway enrichment analyses were conducted for the DEGs using DAVID. Finally, stochastic perturbation was used to determine the significantly differential functions between Gram-positive and Gram-negative samples. A total of 340 and 485 DEGs were obtained in Gram-positive and Gram-negative samples, respectively. Hierarchical clustering revealed that there were significant differences between control and sepsis samples. In Gram-positive and Gram-negative samples, myeloid cell leukemia sequence 1 was associated with apoptosis and programmed cell death. Additionally, NADH:ubiquinone oxidoreductase subunit S4 was associated with mitochondrial respiratory chain complex I assembly. Stochastic perturbation analysis revealed that NADH:ubiquinone oxidoreductase subunit B2 (NDUFB2), NDUFB8 and ubiquinol-cytochrome c reductase hinge protein (UQCRH) were associated with cellular respiration in Gram-negative samples, whereas large tumor suppressor kinase 2 (LATS2) was associated with G1/S transition of the mitotic cell cycle in Gram-positive samples. NDUFB2, NDUFB8 and UQCRH may be biomarkers for Gram-negative sepsis, whereas LATS2 may be a biomarker for Gram-positive sepsis. These findings may promote the therapies of sepsis caused by Gram-positive and Gram-negative bacteria. PMID:28714002

Isolation of Cryptococcus neoformans var. neoformans from bird droppings, fruits and vegetables in Mexico City.

PubMed

López-Martínez, R; Castañón-Olivares, L R

1995-01-01

The presence of Cryptococcus neoformans in various natural sources, such as bird droppings, fruits and vegetables, was investigated. A total of 711 samples were analyzed; C. neoformans var. neoformans was isolated from seven out of 74 bird droppings (9.5%), with parrots as one of the most significant sources. Fruits were positive in 9.5% of the 169 samples studied, specially citrus fruits, particularly grapefruit, in which the highest frequency was found. From the 468 vegetable samples, only 20 were positive (4.2%). It is emphasized that five of the positive vegetables species are autochthonous to Mexico: avocado (Nectandra salicifolia), beet (Beta vulgaris var. quinopodiace), chayote (Sechium edule), stringbean (Cassia sp), and nopal (Opuntia ficus-indica).

[Determination of podophyllotoxin and total lignans in Sinopodophyllum emodi].

PubMed

Huang, Kun; Jiang, Wei; Zhao, Jifeng; Wang, Changhua; Liu, Xiang; Zhang, Zhiwei; Qin, Songyun; Zhong, Guoyue

2012-05-01

To determine the content of podophyllotoxin and total lignans in Sinopodophyllum emodi Ying from different areas and evaluate the resource utilization of this endangered medicinal plant. HPLC and UV spectrophotometry were used to determine the content of podophyllotoxin and total lignans in 126 samples from different habitats and the total lignans content was determined by the reference wavelength method. According to the results, the highest content was determined from the samples from Yongdeng Nature Reserve in Gansu province, and the lowest ones was found in the samples from Tibet. The former's podophyllotoxin and total lignans contents were 7.40% and 20.66%, respectively, which were 19 times and 4 times more than those of the latter. The content of podophyllotoxin and total lignans in S. emodi were significantly positively related, meanwhile, samples from both low altitude and high latitude showed the higher content. The two determination methods are simple, rapid, accurate and repeatable. It is more scientific and rational to evaluate the resource utilization of S. emodi with two indicators, those are the content of podophyllotoxin and the content of total lignans. This paper is instructive to the collection of wild resources and the establishment of production bases.

Discordant HIV Test Results: Implications on Perinatal and Haemotransfusion Screening for HIV Infection, Cape Coast, Ghana.

PubMed

Tetteh, Ato Kwamena; Agyarko, Edward

2017-01-01

Screening results of 488 pregnant women aged 15-44 years whose blood samples had been tested on-site, using First Response® HIV 1/2, and confirmed with INNO-LIA™ HIV I/II Score were used. Of this total, 178 were reactive (HIV I, 154; HIV II, 2; and HIV I and HIV II, 22). Of the 154 HIV I-reactive samples, 104 were confirmed to be HIV I-positive and 2 were confirmed to be HIV II-positive, while 48 were confirmed to be negative [false positive rate = 17.44% (13.56-21.32)]. The two HIV II samples submitted were confirmed to be negative with the confirmatory test. For the 22 HIV I and HIV II samples, 7 were confirmed to be HIV I-positive and 1 was confirmed to be HIV I- and HIV II-positive, while 14 were confirmed to be negative. Of the 310 nonreactive samples, 6 were confirmed to be HIV I-positive and 1 was confirmed to be HIV II-positive [false negative rate = 5.79% (1.63-8.38)], while 303 were negative. False negative outcomes will remain unconfirmed, with no management options for the client. False negative rate of 5.79% requires attention, as its resultant implications on control of HIV/AIDS could be dire.

Collaborative ring-trial of Dynabeads anti-Salmonella for immunomagnetic separation of stressed Salmonella cells from herbs and spices.

PubMed

Mansfield, L; Forsythe, S

1996-02-01

Eight laboratories participated in a Salmonella detection ring-trial which compared selective enrichment by conventional broths with immunomagnetic separation (IMS) using Dynabeads Anti-Salmonella. Laboratories analyzed six types of herbs and spices that were spiked with one of six freeze-dried Salmonella species. Each herb and spice analysis comprised of 12 samples (25 g each) which had been spiked at three different levels, plus a negative control and stored for one week prior to testing. Out of a total 468 samples analyzed, 195 (41.7%) were positive by both methods. Eighteen samples were positive only by IMS enrichment, in comparison with 19 positive samples by conventional enrichment broths and not IMS. These results confirm the potential use of IMS as an alternative to enrichment broths for Salmonella isolation.

Occurency of Giardia duodenalis assemblages in river water sources of Black Sea, Turkey.

PubMed

Koloren, Zeynep; Seferoğlu, Onuralp; Karanis, Panagiotis

2016-12-01

A total of 420 environmental water samples and 120 drinking water samples from 45 different sampling sites of the Black Sea in Turkey were collected between 2012 and 2014. Genomic DNA was isolated from all the investigated water samples and comparativelly analyzed by Loop-mediated isothermal amplification (LAMP) of the elongation factor 1 Alfa (EF1α) gene, and by nested Polymerase Chain Reaction (nPCR) of the small subunit (SSU) rRNA and semi-nested PCR (snPCR) of the glutamate dehydrogenase gene (GDH). 141 (58.7%), 125 (52.1%) and 120 (50%) samples respectivelly were positive by each method. Out of 240 environmental samples collected from 25 sites of Samsun Province have been found positive for G. duodenalis by LAMP, nPCR and snPCR, respectively. 55 (30.5%), 50 (27.8%) and 47 (26.1%) of 180 environmental samples collected from 20 other sampling sites of Giresun Province were positive for Giardia by LAMP, nPCR and snPCR, respectively. Five PCR products from different samples of the Giresun Province and 10 other samples from the Samsun Province were found positive for G. duodenalis assemblage B. Five PCR products from Giresun Province and 5 samples from Samsun Province were found positive for G. duodenalis assemblage A. This is the first report about G. duodenalis assemblages A and B from water samples investigations in Black Sea of Turkey. Copyright © 2016 Elsevier B.V. All rights reserved.

The effects of Campylobacter numbers in caeca on the contamination of broiler carcasses with Campylobacter.

PubMed

Reich, Felix; Atanassova, Viktoria; Haunhorst, Eberhard; Klein, Günter

2008-09-30

For the presence and number of Campylobacter, 18 broiler flocks were sampled over a period of 18 months. A total of 70% of the flocks were positive for Campylobacter, with higher prevalence found in summer and autumn, compared to winter and spring. Positive flocks showed contamination rates above 90%, in negative flocks this was lower, mostly below 50%. The enumeration showed a decrease in Campylobacter during processing of positive flocks. The numbers were highest in carcasses after scalding/defeathering (mean 5.9 log10 cfu/carcass) and dropped by 0.7 log10 cfu/carcass after chilling. A positive correlation was observed between the number of Campylobacter present in the caeca and the number of bacteria present on carcasses and cut products. When a negative flock was slaughtered after Campylobacter positive flocks, the number of positive samples was higher compared to the case when a negative flock had been slaughtered previously. C. jejuni was isolated from 73.6% of the poultry samples.

Identification and determination of the viability of Giardia lamblia cysts and Cryptosporidium parvum and Cryptosporidium hominis oocysts in human fecal and water supply samples by fluorescent in situ hybridization (FISH) and monoclonal antibodies.

PubMed

Lemos, Vanessa; Graczyk, Thaddeus K; Alves, Margarida; Lobo, Maria Luísa; Sousa, Maria C; Antunes, Francisco; Matos, Olga

2005-12-01

In the present study, fluorescent in situ hybridization (FISH) and monoclonal antibodies (MAbs) were evaluated for species-specific detection and viability determination of Giardia lamblia, Cryptosporidium parvum, and Cryptosporidium hominis in human fecal and water supply samples. A total of 50 fecal human samples positive for G. lamblia cysts, 38 positive for C. parvum, and 23 positive for C. hominis were studied. Also, 18 water supply samples positive for Giardia spp. and Cryptosporidium spp. by the United States Environmental Protection Agency (USEPA) Method 1623 were studied by FISH and fluorescein isothiocyanate (FITC)-conjugated MAbs. Eighteen percent of the fecal samples parasitologically positive for G. lamblia presented viable and nonviable cysts, and 5% of those positive for Cryptosporidium spp. presented viable and nonviable oocysts. Of the 18 water supply samples analyzed, 6 (33%) presented Giardia spp. viable and nonviable cysts and 2 (11%) presented viable and nonviable Cryptosporidium spp. oocysts. G. lamblia identification was confirmed by polymerase chain reaction (PCR) and sequencing of the beta-giardin gene in the fecal and water samples found positive by FISH and FITC-conjugated MAbs. C. parvum and Cryptosporidium muris were identified, by PCR and sequencing of the small subunit of ribosomal RNA gene, in seven and one water samples, respectively. Our results confirm that this technique enables simultaneous visualization, species-specific identification, and viability determination of the organisms present in human fecal and water supply samples.

Comparison of ELISA and Microscopy for detection of Cryptosporidium in stool

PubMed Central

Sharma, Madhu; Chaudhary, Uma; Yadav, Aparna

2014-01-01

Background: Cryptosporidiosis, a diarrheal disease caused by the protozoan parasite Cryptosporidium spp. has become recognized as one of the most common causes of water borne diseases in humans. Aims and Objectives: To compare the sensitivity of ELISA and Microscopy for detection of Cryptosporidium in stool samples Materials and Methods: The study was conducted in the Department of Microbiology of PT. B.D. Sharma PGIMS Rohtak, between January 2011 to june 2011 on 50 stool samples, which were processed for detection of cryptosporidial antigen by ELISA and detection of cysts by microscopy (Modified Ziehl and Nelsen staining). Study and Design: This was a prospective study conducted in the Department of Microbiology in PT. BD Sharma, PGIMS, Rohtak, India. Result: Out of total, 50 stool samples eighteen (36%) samples were found positive for Cryptosporidium cysts by microscopy in comparison to 3(6%) stool samples which were found positive for cryptosporidial antigen by ELISA. Samples found positive with ELISA were also positive with microscopy. Sensitivity, specificity, positive predictive value and negative predictive value for ELISA was 16.7%, 100%, 100% and 68% respectively. Conclusion: The study concludes that stool microscopic Modified acid fast staining is more sensitive method than ELISA for detection of Cryptosporidium in stool samples but the specificity of ELISA was more than microscopy. PMID:25584216

Serological diagnosis of bovine brucellosis using B. melitensis strain B115.

PubMed

Corrente, Marialaura; Desario, Costantina; Parisi, Antonio; Grandolfo, Erika; Scaltrito, Domenico; Vesco, Gesualdo; Colao, Valeriana; Buonavoglia, Domenico

2015-12-01

Bovine brucellosis is diagnosed by official tests, such as Rose Bengal plate test (RBPT) and Complement Fixation test (CFT). Both tests detect antibodies directed against the lipolysaccharide (LPS) of Brucella cell wall. Despite their good sensitivity, those tests do not discriminate between true positive and false positive serological reactions (FPSR), the latter being generated by animals infected with other Gram negative microorganisms that share components of Brucella LPS. In this study, an antigenic extract from whole Brucella melitensis B115 strain was used to set up an ELISA assay for the serological diagnosis of bovine brucellosis. A total of 148 serum samples from five different groups of animals were tested: Group A: 28 samples from two calves experimentally infected with Yersinia enterocolitica O:9; Group B: 30 samples from bovines infected with Brucella abortus; Group C: 50 samples from brucellosis-free herds; Group D: 20 samples RBPT positive and CFT negative; Group E: 20 samples both RBPT and CFT positive. Group D and Group E serum samples were from brucellosis-free herds. Positive reactions were detected only by RBPT and CFT in calves immunized with Y. enterocolitica O:9. Sera from Group B animals tested positive also in the ELISA assay, whereas sera from the remaining groups were all negative. The results obtained encourage the use of the ELISA assay to implement the serological diagnosis of brucellosis. Copyright © 2015 Elsevier B.V. All rights reserved.

Groundwater microbiological quality in Canadian drinking water municipal wells.

PubMed

Locas, Annie; Barthe, Christine; Margolin, Aaron B; Payment, Pierre

2008-06-01

To verify previous conclusions on the use of bacterial indicators suggested in regulations and to investigate virological quality of groundwater, a 1-year study was undertaken on groundwater used as a source of drinking water in 3 provinces in Canada. Raw water from 25 municipal wells was sampled during a 1-year period for a total of 167 samples. Twenty-three sites were selected on the basis of their excellent historical bacteriological water quality data, and 2 sites with known bacteriological contamination were selected as positive controls. Water samples were analyzed for general water quality indicators (aerobic endospores, total coliforms), fecal indicators (Escherichia coli, enterococci, somatic and male-specific coliphages), total culturable human enteric viruses (determined by cell culture and immunoperoxidase), noroviruses (analyzed by reverse-transcriptase -- polymerase chain reaction (RT-PCR)), adenovirus types 40 and 41 (analyzed by integrated cell culture (ICC) - PCR), and enteroviruses and reoviruses types 1, 2, and 3 (analyzed by ICC-RT-PCR). General water quality indicators were found very occasionally at the clean sites but were frequently present at the 2 contaminated sites. Only one of 129 samples from the 23 clean sites was positive for enterococci. These results confirm the value of raw water quality historical data to detect source water contamination affecting wells that are vulnerable. Samples from the 2 contaminated sites confirmed the frequent presence of fecal indicators: E. coli was found in 20/38 samples and enterococci in 12/38 samples. Human enteric viruses were not detected by cell culture on MA-104 cells nor by immunoperoxidase detection in any sample from the clean sites but were found at one contaminated site. By ICC-RT-PCR and ICC-PCR, viruses were found by cytopathic effect in one sample from a clean site and they were found in 3 samples from contaminated sites. The viruses were not detected by the molecular methods but were confirmed as picornaviruses by electron microscopy. Noroviruses were not detected in any samples. The results obtained reinforce the value of frequent sampling of raw water using simple parameters: sampling for total coliforms and E. coli remains the best approach to detect contamination of source water by fecal pollutants and accompanying pathogens. The absence of total coliforms at a site appears to be a good indication of the absence of human enteric viruses.

A study of the dynamics of Salmonella infection in turkey breeding, rearing and finishing houses with special reference to elimination, persistence and introduction of Salmonella.

PubMed

Mueller-Doblies, Doris; Carrique-Mas, Juan J; Davies, Robert H

2014-01-01

In this descriptive study, the dynamics of Salmonella infection of turkey flocks were investigated by repeated sampling of houses where Salmonella had been identified. The aim of the study was to identify the most common scenarios involved in elimination, persistence and introduction of Salmonella in the different branches of the turkey industry. Sixty-two houses on 34 turkey farms (comprising breeding, rearing and finishing farms) were sampled longitudinally, starting with the identification of a positive flock. A total of 117 follow-on flocks were tested and cleaning and disinfection (C&D) was assessed during 66 post-C&D visits. A total of 155 incidents (clearance, persistence or introduction of Salmonella) were recorded. Persistence was seen in 35.5% of incidents and was seen more frequently in breeding and rearing houses compared with finishing houses. Most persistence incidents were the result of insufficient C&D. Clearance was seen in 40% of incidents and was more often observed in finishing houses than in breeding or rearing houses. Introduction was seen in 24.5% of incidents and was more common in breeding and finishing flocks than in rearing flocks. Contamination of a house with Salmonella Typhimurium was more likely to be cleared compared with other serovars. The total number of positive samples found at a post-C&D visit was correlated with the probability of carry-over of infection, whereas the location of the positive samples seemed to be less important. Our highly sensitive post-C&D sampling method allowed us to predict a negative follow-on flock in most cases.

Knowledge and attitude towards total knee arthroplasty among the public in Saudi Arabia: a nationwide population-based study.

PubMed

Al-Mohrej, Omar A; Alshammari, Faris O; Aljuraisi, Abdulrahman M; Bin Amer, Lujain A; Masuadi, Emad M; Al-Kenani, Nader S

2018-04-01

Studies on total knee arthroplasty (TKA) in Saudi Arabia are scarce, and none have reported the knowledge and attitude of the procedure in Saudi Arabia. Our study aims to measure the knowledge and attitude of TKA among the adult Saudi population. To encompass a representative sample of this cross-sectional survey, all 13 administrative areas were used as ready-made geographical clusters. For each cluster, stratified random sampling was performed to maximize participation in the study. In each area, random samples of mobile phone numbers were selected with a probability proportional to the administrative area population size. Sample size calculation was based on the assumption that 50% of the participants would have some level of knowledge, with a 2% margin of error and 95% confidence level. To reach our intended sample size of 1540, we contacted 1722 participants with a response rate of 89.4%. The expected percentage of public knowledge was 50%; however, the actual percentage revealed by this study was much lower (29.7%). A stepwise multiple logistic regression was used to assess the factors that positively affected the knowledge score regarding TKA. Age [P = 0.016 with OR of 0.47], higher income [P = 0.001 with OR of 0.52] and participants with a positive history of TKA or that have known someone who underwent the surgery [P < 0.001 with OR of 0.15] had a positive impact on the total knowledge score. There are still misconceptions among the public in Saudi Arabia concerning TKA, its indications and results. We recommend that doctors use the results of our survey to assess their conversations with their patients, and to determine whether the results of the procedure are adequately clarified.

The relationship between maternal attachment, perceived social support and breast-feeding sufficiency.

PubMed

Cinar, Nursan; Köse, Dilek; Altinkaynak, Sevin

2015-04-01

To determine the relationship between maternal attachment, perceived social support and breast-feeding sufficiency. Descriptive correlational design. A state hospital and two family health centers in Sakarya, Turkey, between June to December 2011. The sample was 122 voluntary mothers who had healthy babies of 1 - 2 months old. The data were collected by a Personal Information Form, Maternal Attachment Inventory (MAI), Multidimensional Scale of Perceived Social Support (MSPSS) and Breast-feeding Self-Efficacy Scale-Short Form (BSES-SF). The data collected were analysed by percentage distribution, mean square, independent sample t-test, Mann-Whitney U, Kruskall-Wallis and Pearson correlation. The mean age of the mothers was 25.01 ± 2.2 years, and 48.4% of them were primary school graduates. BSESSF was 61.02 ± 8.44 (16 - 70), MAI was 99.07 ± 7.19 (56 - 100) and MSPSS was 66.40 ± 13.58 (37 - 84). There was a positive, medium level, significant relationship between the total scores of BSES-SF and MAI (r=0.370, p < 0.001). There was a positive, medium level, significant relationship between the total score of BSES-SF and the score from family subdimension of MSPSS (r=0.255, p < 0.01).There was a positive, medium level, significant relationship between the total score of MAI and the total score of family subdimension of MSPSS (r=0.339, p < 0.001). Successful maternal attachment and familial support positively affected the breast-feeding sufficiency of the mother.

Density of total and pathogenic (tdh+) Vibrio parahaemolyticus in Atlantic and Gulf coast molluscan shellfish at harvest.

PubMed

Cook, David W; Bowers, John C; DePaola, Angelo

2002-12-01

The densities of total and pathogenic Vibrio parahaemolyticus in 671 samples of molluscan shellfish harvested in 1999 and 2000 from 14 sites in seven Gulf and Atlantic coast states were determined at 2-week intervals over a period of 12 to 16 months in each state. Changes in V. parahaemolyticus densities in shellfish between harvest and sample analysis were minimized with time and temperature controls. Densities were measured by direct plating techniques, and gene probes were used for identification. Total and pathogenic V. parahaemolyticus organisms were identified with probes for the thermolabile direct hemolysin (tlh) gene and the thermostable direct hemolysin (tdh) gene, respectively. An enrichment procedure involving 25 g of shellfish was also used for the recovery of pathogenic V. parahaemolyticus. The densities of V. parahaemolyticus in shellfish from all harvest sites were positively correlated with water temperature. Shellfish from the Gulf Coast typically had higher densities of V. parahaemolyticus than did shellfish harvested from the North Atlantic or mid-Atlantic coast. Vibrio parahaemolyticus counts exceeded 1,000 CFU/g for only 5% of all samples. Pathogenic (tdh+) V. parahaemolyticus was detected in approximately 6% of all samples by both procedures, and 61.5% of populations in the positive samples from the direct plating procedure were at the lower limit of detection (10 CFU/g). The frequency of detection of pathogenic V. parahaemolyticus was significantly related to water temperature and to the density of total V. parahaemolyticus. The failure to detect pathogenic V. parahaemolyticus in shellfish more frequently was attributed to the low numbers and uneven distribution of the organism.

[Characteristics of blood type irregular antibodies in Han population of Chinese Sichuan area].

PubMed

Li, Cui-Ying; Li, Yun-Ming; Huang, Fei; Xiao, Jie; Xu, Hong

2015-04-01

To analyze the distribution of irregular antibody of red blood cells in Han population of Chinese Sichuan area, so as to provide valuable information for the safety of transfusion and decrease of immune hemolytic transfusion reaction. Blood samples from June 2006 to May 2013 were tested for irregular antibody screening and identification, calculating the composition rate, group characteristics and the positive detection rate of irregular antibody. A total of 36287 blood samples were tested, out of them 571 samples were the irregular antibody positive, the positive rate was 1.574%(571/36 287), specific alloantibodies were found in 312 samples, the positive rate was 0.860%(312/36287). And autoantibody was found in 259 samples, the positive rate was 0.714%(259/36 287). The specific alloantibodies ratio in Rh system was the highest, reaching to 73.72%(230/312) with the positive rate of 0.634%;36 cases in Lewis system, account for 11.54%(36/312) with the positive rate of 0.099%; 34 cases in MNS system account for 10.89%(34/312) with the positive rate of 0.094%; direct coomb test showed positive result in 284 samples, the rate was 0.78%. The detected rate of positive irregular antibody in female is obviously higher than that in male patients (P<0.001), and it is also higher in people with pregnancy or transfusion than that in those without it (P<0.05). The irregular antibody screening and identification are very important in blood transfusion, especially for female and people with transfusion or pregnant history.

Should gram stains have a role in diagnosing hip arthroplasty infections?

PubMed

Johnson, Aaron J; Zywiel, Michael G; Stroh, D Alex; Marker, David R; Mont, Michael A

2010-09-01

The utility of Gram stains in diagnosing periprosthetic infections following total hip arthroplasty has recently been questioned. Several studies report low sensitivity of the test, and its poor ability to either confirm or rule out infection in patients undergoing revision total hip arthroplasty. Despite this, many institutions including that of the senior author continue to perform Gram stains during revision total hip arthroplasty. We assessed the sensitivity, specificity, accuracy, and positive and negative predictive values of Gram stains from surgical-site samplings taken from procedures on patients with both infected and aseptic revision total hip arthroplasties. A review was performed on patients who underwent revision total hip arthroplasty between 2000 and 2007. Eighty-two Gram stains were performed on patients who had infected total hip arthroplasties and underwent revision procedures. Additionally, of the 410 revision total hip arthroplasties performed on patients who were confirmed infection-free, 120 Gram stains were performed. Patients were diagnosed as infected using multiple criteria at the time of surgery. Sensitivity, specificity, positive and negative predictive values, and accuracy were calculated from these Gram stain results. The Gram stain demonstrated a sensitivity and specificity of 9.8% and 100%, respectively. In this series, the Gram stain had a negative predictive value of 62%, a positive predictive value of 100%, and an accuracy of 63%. Gram stains obtained from surgical-site samples had poor sensitivity and poor negative predictive value. Based on these findings, as well as those of other authors, we believe that Gram stains should no longer be considered for diagnosing infections in revision total hip arthroplasty. Level III, diagnostic study. See Guidelines for Authors for a complete description of levels of evidence.

Differences in soil biological activity by terrain types at the sub-field scale in central Iowa US

DOE PAGES

Kaleita, Amy L.; Schott, Linda R.; Hargreaves, Sarah K.; ...

2017-07-07

Soil microbial communities are structured by biogeochemical processes that occur at many different spatial scales, which makes soil sampling difficult. Because soil microbial communities are important in nutrient cycling and soil fertility, it is important to understand how microbial communities function within the heterogeneous soil landscape. In this study, a self-organizing map was used to determine whether landscape data can be used to characterize the distribution of microbial biomass and activity in order to provide an improved understanding of soil microbial community function. Points within a row crop field in south-central Iowa were clustered via a self-organizing map using sixmore » landscape properties into three separate landscape clusters. Twelve sampling locations per cluster were chosen for a total of 36 locations. After the soil samples were collected, the samples were then analysed for various metabolic indicators, such as nitrogen and carbon mineralization, extractable organic carbon, microbial biomass, etc. It was found that sampling locations located in the potholes and toe slope positions had significantly greater microbial biomass nitrogen and carbon, total carbon, total nitrogen and extractable organic carbon than the other two landscape position clusters, while locations located on the upslope did not differ significantly from the other landscape clusters. However, factors such as nitrate, ammonia, and nitrogen and carbon mineralization did not differ significantly across the landscape. Altogether, this research demonstrates the effectiveness of a terrain-based clustering method for guiding soil sampling of microbial communities.« less

Differences in soil biological activity by terrain types at the sub-field scale in central Iowa US

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kaleita, Amy L.; Schott, Linda R.; Hargreaves, Sarah K.

Soil microbial communities are structured by biogeochemical processes that occur at many different spatial scales, which makes soil sampling difficult. Because soil microbial communities are important in nutrient cycling and soil fertility, it is important to understand how microbial communities function within the heterogeneous soil landscape. In this study, a self-organizing map was used to determine whether landscape data can be used to characterize the distribution of microbial biomass and activity in order to provide an improved understanding of soil microbial community function. Points within a row crop field in south-central Iowa were clustered via a self-organizing map using sixmore » landscape properties into three separate landscape clusters. Twelve sampling locations per cluster were chosen for a total of 36 locations. After the soil samples were collected, the samples were then analysed for various metabolic indicators, such as nitrogen and carbon mineralization, extractable organic carbon, microbial biomass, etc. It was found that sampling locations located in the potholes and toe slope positions had significantly greater microbial biomass nitrogen and carbon, total carbon, total nitrogen and extractable organic carbon than the other two landscape position clusters, while locations located on the upslope did not differ significantly from the other landscape clusters. However, factors such as nitrate, ammonia, and nitrogen and carbon mineralization did not differ significantly across the landscape. Altogether, this research demonstrates the effectiveness of a terrain-based clustering method for guiding soil sampling of microbial communities.« less

High prevalence of Legionella in non-passenger merchant vessels.

PubMed

Collins, S L; Stevenson, D; Mentasti, M; Shaw, A; Johnson, A; Crossley, L; Willis, C

2017-03-01

There is a paucity of information on the risk from potable water in non-passenger merchant vessels (NPMVs) particularly with regard to Legionella and other bacteria. This retrospective study examined water samples from 550 NPMVs docked in eight UK ports. A total of 1027 samples from 412 NPMVs were examined for total aerobic colony counts (ACC), coliforms, Escherichia coli and enterococci; 41% of samples yielded ACC above the action level (>1 × 103 c.f.u./ml) and 4·5% contained actionable levels (>1 c.f.u./100 ml) of faecal indicator bacteria. Eight hundred and three samples from 360 NPMVs were cultured specifically for Legionella and 58% of vessels proved positive for these organisms with 27% of samples showing levels greater than the UK upper action limit of 1 × 103 c.f.u./l. Cabin showers (49%) and hospital shower (45%) were frequently positive. A subset of 106 samples was analysed by quantitative polymerase chain reaction for Legionella and identified a further 11 Legionella-positive NPMVs, returning a negative predictive value of 100%. There was no correlation between NPMV age or size and any microbial parameters (P > 0·05). Legionella pneumophila serogroup 1 was isolated from 46% of NPMVs and sequence-based typing of 17 isolates revealed four sequence types (STs) previously associated with human disease. These data raise significant concerns regarding the management of microbial and Legionella risks on board NPMVs and suggest that better guidance and compliance are required to improve control.

40 CFR 141.857 - Routine monitoring requirements for public water systems serving more than 1,000 people.

Code of Federal Regulations, 2013 CFR

2013-07-01

... REGULATIONS Revised Total Coliform Rule § 141.857 Routine monitoring requirements for public water systems... systems serving more than 1,000 persons. (2) Following any total coliform-positive sample taken under the... calendar month has been completed, systems must determine whether any coliform treatment technique triggers...

Subcortical intelligence: caudate volume predicts IQ in healthy adults.

PubMed

Grazioplene, Rachael G; G Ryman, Sephira; Gray, Jeremy R; Rustichini, Aldo; Jung, Rex E; DeYoung, Colin G

2015-04-01

This study examined the association between size of the caudate nuclei and intelligence. Based on the central role of the caudate in learning, as well as neuroimaging studies linking greater caudate volume to better attentional function, verbal ability, and dopamine receptor availability, we hypothesized the existence of a positive association between intelligence and caudate volume in three large independent samples of healthy adults (total N = 517). Regression of IQ onto bilateral caudate volume controlling for age, sex, and total brain volume indicated a significant positive correlation between caudate volume and intelligence, with a comparable magnitude of effect across each of the three samples. No other subcortical structures were independently associated with IQ, suggesting a specific biological link between caudate morphology and intelligence. © 2014 Wiley Periodicals, Inc.

Prevalence of doping in sports: doping control in Norway, 1977-1995.

PubMed

Bahr, R; Tjørnhom, M

1998-01-01

To examine the results from doping controls conducted by the Norwegian Confederation of Sport (NCS) from 1977 to 1995. Data were collected by combining three computerized databases and manual records on samples taken and results from analyses in the International Olympic Committee (IOC)-accredited laboratories in London, Huddinge, Cologne, and Oslo. Samples were declared positive if they contained any banned substance on the IOC list that was in effect at any given time. A total of 15,208 samples were taken; most of them (12,870; 85%) were from Norwegian athletes (90% unannounced tests) belonging to national federations under NCS jurisdiction (NCS members), 461 (3%) were from external Norwegian athletes (either users of private gyms or athletes in organized sports federations not affiliated with the NCS), and 1,874 (12%) were from foreign athletes (three cases with unknown affiliation). There were 130 positive samples and 24 refusals among NCS members (1.2%; men, 1.4%; women, 0.3%), 86 positive samples and 8 refusals among external Norwegian athletes (20%; men, 24%; women, 8%), and 39 positive samples and 1 refusal among foreign athletes (1.6%; men, 2.1%; women, 0.7%). A gradual decrease in the percentage of positive samples was observed among NCS members as testing frequency was increased gradually from 1987 to 1995 in the three high-prevalence sports: powerlifting, weightlifting, and athletics. An increase in the test frequency of doping tests was associated with a decrease in the percentage of positive samples in targeted sports.

Detection of Salmonellae in the Environment

PubMed Central

Thomason, Berenice M.; Biddle, James W.; Cherry, William B.

1975-01-01

The incidence of salmonellae in contrasting environments was compared in this study. Samples collected from or near surface waters in a lush hardwood forest yielded four salmonellae serotypes from six culturally positive samples. A total of 76 samples collected from the top of a granite outcropping over a 3-month period yielded 10 positive samples. Only two salmonellae serotypes were isolated, and one of these was isolated only once. The nature of the sample material had no significant effect on the detection of salmonellae from the two sampling sites. However, the presence or absence of visible moisture in the sample significantly affected the recovery of salmonellae. The results showed that even a harsh environment such as that found on top of Stone Mountain may serve as an ecological niche for the survival and transmission of salmonellae. PMID:1106319

Performance of VITEK® MS V3.0 for the Identification of Mycobacterium species from Patient Samples using Automated Liquid Media Systems.

PubMed

Miller, Eric; Cantrell, Christopher; Beard, Melodie; Derylak, Andrew; Babady, N Esther; McMillen, Tracy; Miranda, Edwin; Body, Barbara; Tang, Yi-Wei; Vasireddy, Ravikiran; Vasireddy, Sruthi; Smith, Terry; Iakhiaeva, Elena; Wallace, Richard J; Brown-Elliott, Barbara A; Moreno, Erik; Totty, Heather; Deol, Parampal

2018-06-06

The accuracy and robustness of the VITEK® MS V3.0 matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) system was evaluated by identifying mycobacteria from automated liquid media systems using patient samples. This is the first report demonstrating that proteins within the liquid media, its supplements, and decontamination reagents for non-sterile patient samples do not generate misidentification or false positive results when using the VITEK MS V3.0 system. Prior to testing with patient samples, a seeded study was conducted to challenge the accuracy of the VITEK MS to identify mycobacteria from liquid media through mimicking a clinical workflow. A total of 77 Mycobacterium strains representing 21 species, seeded in simulated sputum, were decontaminated and inoculated into BACT/ALERT®MP liquid culture medium, incubated until positivity, and identified using VITEK MS. A total of 383 liquid cultures were tested of which 379 (99%) identified correctly to the species/complex/group; four (1%) obtained a No Identification, and no misidentifications were observed. Following the simulated sputum study, a total of 73 smear-positive liquid medium cultures detected using BD BBL™ MGIT™ and VersaTREK® MYCO liquid media were identified by VITEK MS. Sixty-four (87.7%) correctly identified to the species/complex/group level; seven (9.6%) resulted as No Identification, and two (2.7%) misidentified at the species level. These results indicate the VITEK MS V3.0 is an accurate tool for routine diagnostics of Mycobacterium species isolated from liquid cultures. Copyright © 2018 American Society for Microbiology.

Assessment of mercury exposure among small-scale gold miners using mercury stable isotopes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sherman, Laura S., E-mail: lsaylors@umich.edu; Blum, Joel D.; Basu, Niladri

Total mercury (Hg) concentrations in hair and urine are often used as biomarkers of exposure to fish-derived methylmercury (MeHg) and gaseous elemental Hg, respectively. We used Hg stable isotopes to assess the validity of these biomarkers among small-scale gold mining populations in Ghana and Indonesia. Urine from Ghanaian miners displayed similar Δ{sup 199}Hg values to Hg derived from ore deposits (mean urine Δ{sup 199}Hg=0.01‰, n=6). This suggests that urine total Hg concentrations accurately reflect exposure to inorganic Hg among this population. Hair samples from Ghanaian miners displayed low positive Δ{sup 199}Hg values (0.23–0.55‰, n=6) and low percentages of total Hgmore » as MeHg (7.6–29%, n=7). These data suggest that the majority of the Hg in these miners' hair samples is exogenously adsorbed inorganic Hg and not fish-derived MeHg. Hair samples from Indonesian gold miners who eat fish daily displayed a wider range of positive Δ{sup 199}Hg values (0.21–1.32‰, n=5) and percentages of total Hg as MeHg (32–72%, n=4). This suggests that total Hg in the hair samples from Indonesian gold miners is likely a mixture of ingested fish MeHg and exogenously adsorbed inorganic Hg. Based on data from both populations, we suggest that total Hg concentrations in hair samples from small-scale gold miners likely overestimate exposure to MeHg from fish consumption. - Highlights: • Mercury isotopes were measured in hair and urine from small-scale gold miners. • Mercury isotopes indicate that Hg in urine comes from mining activity. • Mercury isotopes suggest Hg in hair is a mixture of fish MeHg and inorganic Hg. • A large percentage of Hg in miner’s hair is released during amalgam burning and adsorbed.« less

Significant increase in cultivation of Gardnerella vaginalis, Alloscardovia omnicolens, Actinotignum schaalii, and Actinomyces spp. in urine samples with total laboratory automation.

PubMed

Klein, Sabrina; Nurjadi, Dennis; Horner, Susanne; Heeg, Klaus; Zimmermann, Stefan; Burckhardt, Irene

2018-04-13

While total laboratory automation (TLA) is well established in laboratory medicine, only a few microbiological laboratories are using TLA systems. Especially in terms of speed and accuracy, working with TLA is expected to be superior to conventional microbiology. We compared in total 35,564 microbiological urine cultures with and without incubation and processing with BD Kiestra TLA for a 6-month period each retrospectively. Sixteen thousand three hundred thirty-eight urine samples were analyzed in the pre-TLA period and 19,226 with TLA. Sixty-two percent (n = 10,101/16338) of the cultures processed without TLA and 68% (n = 13,102/19226) of the cultures processed with TLA showed growth. There were significantly more samples with two or more species per sample and with low numbers of colony forming units (CFU) after incubation with TLA. Regarding the type of bacteria, there were comparable amounts of Enterobacteriaceae in the samples, slightly less non-fermenting Gram-negative bacteria, but significantly more Gram-positive cocci, and Gram-positive rods. Especially Alloscardivia omnicolens, Gardnerella vaginalis, Actinomyces spp., and Actinotignum schaalii were significantly more abundant in the samples incubated and processed with TLA. The time to report was significantly lower in the TLA processed samples by 1.5 h. We provide the first report in Europe of a large number of urine samples processed with TLA. TLA showed enhanced growth of non-classical and rarely cultured bacteria from urine samples. Our findings suggest that previously underestimated bacteria may be relevant pathogens for urinary tract infections. Further studies are needed to confirm our findings.

Prevalence of virulent Yersinia enterocolitica in bulk raw milk and retail cheese in northern-west of Iran.

PubMed

Hanifian, Shahram; Khani, Sajjad

2012-04-02

To determine the prevalence of virulent Yersinia enterocolitica, 554 samples consisting of 354 bulk raw milks and 200 traditional cheeses were collected from different parts of Eastern-Azerbaijan province, during a 23-month period from 2008 to 2010. The occurrence of virulent strains of Y. enterocolitica in samples enriched in peptone sorbitol bile broth (PSBB) was evaluated via the detection of attachment invasion locus (ail) gene by PCR. The viability of virulent Y. enterocolitica in the PCR-positive samples was tested using conventional culture method and the isolates were confirmed by the second-phase ail-PCR. According to the results, 8.66% of total samples including 7.62% of bulk raw milks and 10.5% of raw milk cheeses were found ail-positive by PCR method; subsequently Y. enterocolitica was isolated by the culture method and confirmed by the second phase ail-PCR in 2.88% of total samples including 2.26% of raw milks and 4% of cheese samples. It was concluded that, a sample enrichment followed by ail-PCR was more sensitive and robust to detect and distinguish the virulent strains of Y. enterocolitica compared to the conventional culture method. Copyright © 2012 Elsevier B.V. All rights reserved.

Insight, psychosis, and depression in Africa: a cross-sectional survey from an inpatient unit in Ghana.

PubMed

Poole, Norman A; Crabb, Jim; Osei, Akwasi; Hughes, Peter; Young, David

2013-06-01

Few studies of the relationship of insight to psychopathology have been conducted in non-Western populations. This study examined the relationships between insight and depression, anxiety, and positive and negative symptoms on patients with schizophrenia resident in a psychiatric hospital in Ghana. A sample of 49 participants, (37 men and 12 women), with DSM-IV defined schizophrenia took part in semistructured interviews consisting of the Hamilton Rating Scales for Depression (HAM-D) and Anxiety (HAM-A); the Schedule for the Assessment of Insight - Expanded Version (SAI-E) and the Positive and Negative Syndrome Scale (PANSS). Bivariate correlations between variables were examined and those significantly correlated with an insight domain were included in multiple regression models. Variables associated with the total insight score were age, gender, anxiety symptoms, depression symptoms, and treatment compliance. In the final model, HAM-D positively predicted total SAI-E score, whilst PANSS-pos was negatively associated with total SAI-E score. The results are broadly consistent with those found in Western samples regarding insight and depressive symptoms. Implications of these results for competing theories of insight in psychoses are discussed. Patients able to identify themselves as ill may be aware of their affective symptoms.

Microbial indicators in natural biofilms developed in the riverbed.

PubMed

Hirotani, Hiroshi; Yoshino, Miyuki

2010-01-01

Microbial indicators such as heterotrophic bacteria, total coliforms, and Escherichia coli in naturally developed riverbed biofilms were investigated. Pebbles covered with natural biofilm were sampled directly from the riverbed at sampling stations ranging from the upstream region within a quasi-national park to the midstream in the urban district. Heterotrophic bacteria densities in biofilm positively correlated with stream discharge. E. coli densities in biofilm positively correlated with temperature, which suggests the growth in the biofilm. It was considered that the attachment of planktonic bacteria to biofilm was negligible. The biofilm may serve as an internal source of false positive indication of fecal contamination in the water column.

Natural stable isotopic compositions of mercury in aerosols and wet precipitations around a coal-fired power plant in Xiamen, southeast China

NASA Astrophysics Data System (ADS)

Huang, Shuyuan; Sun, Lumin; Zhou, Tingjin; Yuan, Dongxing; Du, Bing; Sun, Xiuwu

2018-01-01

In this study, samples of 18 wet precipitations (WPs) and 38 aerosols were collected around a coal-fired power plant (CFPP) located in Xiamen, southeast China, which was equipped with a seawater flue gas desulfurization system. Total particulate mercury (TPM) in aerosol samples, and total mercury (WP-TM), dissolved mercury (WP-DM) and particulate mercury (WP-PM) in WP samples were analyzed for the natural isotopic compositions of mercury. For the first time, both mass dependent fractionation (MDF) and mass independent fractionation of odd (odd-MIF) and even (even-MIF) isotopes of WP-DM and WP-PM were reported and discussed. Both WP-TM and TPM displayed negative MDF and slightly positive even-MIF. Negative odd-MIF was observed in TPM and WP-PM, whereas positive odd-MIF was observed in WP-TM and WP-DM. It was found that the mercury budget in WP-PM samples was mainly controlled by atmospheric particles. Potential sources of mercury in samples were identified via analysis of mercury isotopic signatures and meteorological data with the NOAA HYSPLIT model. The results showed that TPM and WP-PM in solid samples were homologous and the isotopic compositions of WP-TM depended on those of WP-DM. The ratios of Δ199Hg/Δ201Hg resulting from photochemical reactions and positive Δ200Hg values (from -0.06‰ to 0.27‰) in all samples indicated that the mercury coming from local emission of the CFPP together with long-distance transportation were the two main contributing sources.

Evaluation of a malarial antibody assay for use in the screening of blood and tissue products for clinical use.

PubMed

Kitchen, A D; Lowe, P H J; Lalloo, K; Chiodini, P L

2004-10-01

A new recombinant Plasmodium antigen enzyme immunoassay (EIA) for the detection of malarial antibodies was evaluated for the screening of 'malaria-risk' blood and tissue donations. A total of 13,269 donor and patient samples were tested by both the EIA and the standard diagnostic antibody immunofluorescence test (IFAT). A total of 114/138 (82.6%) samples from patients with P. falciparum and 11/13 (84.6%) samples from patients with P. vivax tested positive. A total of 714/13,053 (5.47%) samples from donors identified as 'malaria risk', owing to residency or travel, were reactive in the EIA. The assay is more sensitive than a previously implemented malarial antibody EIA (73% in acute P. falciparum and 56% in acute P. vivax infections). The sensitivity of this new EIA is comparable to that of the IFAT, and the specificity is sufficient to screen 'malaria-risk' donors.

[Evaluation of mycobacterial microscopy and culture results of Sureyyapasa Chest Diseases and Chest Surgery Training and Research Hospital: A 3-year analysis].

PubMed

Akduman Alaşehir, Elçin; Balıkçı, Ahmet; Partal, Mualla; Çatmabacak, Gülay; Yaman, Görkem

2016-09-01

Effective diagnosis of tuberculosis is of great importance for transmission control and treatment success. The purpose of this study is to evaluate microscopic examination results of Ehrlich-Ziehl Neelsen (EZN) and Auramine-Rhodamine staining methods and automated BACTEC MGIT 960™ system and Löwenstein-Jensen (L-J) culture results of various clinical samples in the light of recent data from the world and Turkey. Specimens that were sent from various clinics to Sureyyapasa Chest Diseases and Chest Surgery Training and Research Hospital Microbiology Laboratory from January 2012 to December 2015 were evaluated retrospectively. From a total of 62456 samples; 60923 (97.5%) were pulmonary and 1533 (2.5%) were non-pulmonary samples, especially pleura. 2853 (4.6%) Acid-resistant bacilli (ARB) positivity was detected and mycobacterial culture positivity was in total 12.2%. 7076 (93%) and 535 (7%) mycobacteria other than tuberculosis (MOTT) strains were isolated. In 356 specimens the cultures were negative in despite the positive ARB results. Considering mycobacterial culture as the gold standard; the sensitivity, specificity, positive and negative predictive values of ARB microscopy were 32.8%, 99.4%, 87.5% and 91.4%, respectively. The contamination rates in total were within acceptable limits being 2.7% for L-J and 3.8% for MGIT. Analysis of our data indicated that the sensitivity of microscopy is low and it should be evaluated together with the mycobacterial culture to rule out tuberculosis infection. With the use of fluorescent staining and also L-J and MGIT broth together for routine culture since 2013; ARB false negativity rate was observed to fall to 51.7% from 74.1% compared to the years. The follow-up of data such as the sensitivity of microscopy, culture positivity, false-positivity and false-negativity rates and contamination values is of great importance in terms of assessing compliance with laboratory quality standards and contributing to the surveillance studies.

Some blood chemistry values for five Chesapeake Bay area fishes

USGS Publications Warehouse

Hunn, J.B.; Robinson, P.F.

1966-01-01

Blood samples from gizzard shad,largemouth bass, white perch, pumpkinseed, and toadfish were analyzed for hemoglobin, total plasma protein, total plasma cholesterol, and ion concentrations of plasma sodium, potassium, and chloride. The hemoglobin concentration and total plasma cholesterol found in a given species seem to have positive correlation with the customary activity level of that species. The plasma ionic concentrations in general agree with those found by other authors.

Diagnostic usefulness of Xpert MTB/RIF assay for detection of tuberculous meningitis using cerebrospinal fluid.

PubMed

Rufai, Syed Beenish; Singh, Amit; Singh, Jitendra; Kumar, Parveen; Sankar, Manimuthu Mani; Singh, Sarman

2017-08-01

Tuberculous meningitis (TBM) is the most severe form of extra-pulmonary tuberculosis (TB) due to association of diseases with high rates of mortality and morbidity. Diagnosis continues to be a clinical challenge as microbiological confirmation is rare and time consuming resulting in delayed treatment. Xpert MTB/RIF assay is a rapid and simple test, which has been endorsed by World Health Organization as an initial diagnostic test for the diagnosis of TBM. However, evidence still lacks for its performance on cerebrospinal fluid (CSF) for the diagnosis of TBM especially from India. A total of 267 CSF samples from patients with high clinico-radiological suspicion of TBM were included in this study. Ziehl-Neelsen (ZN) staining, BACTEC Mycobacterial Growth Indicator Tube (MGIT-960) culture system, and Xpert MTB/RIF assay (using cartridge version G4) were tested on all samples. Of total 267 samples, all were negative for smear AFB and 52 (19.5%) were culture positive by MGIT-960 culture system. However, out of 52 (19.5%) cultures detected positive by MGIT-960, 5 (9.6%) were detected as resistant to rifampicin. Xpert MTB/RIF assay was positive in 38 (14.2%) samples and negative in 223 (83.5%) samples. Cartridge error was detected in 6 (2.2%) samples, which could not be repeated due to insufficient sample volume. The sensitivity and specificity of Xpert MTB/RIF assay in comparison to MGIT-960 was 55.1% (95%, CI: 40.2-69.3) and 94.8% (95%, CI: 90.9-97.4) respectively. Overall, Xpert MTB/RIF assay detected 38 (14.2%) as positive for MTB of which 4 (10.5%), 31 (81.6%) and 3 (7.9%) were found to be rifampicin resistant, sensitive and indeterminate respectively. Xpert MTB/RIF assay showed lower sensitivity as compared to MGIT 960 culture for the diagnosis of TBM from CSF samples. Copyright © 2017 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

Ubiquity of microplastics in coastal seafloor sediments.

PubMed

Ling, S D; Sinclair, M; Levi, C J; Reeves, S E; Edgar, G J

2017-08-15

Microplastic pollutants occur in marine environments globally, however estimates of seafloor concentrations are rare. Here we apply a novel method to quantify size-graded (0.038-4.0mm diam.) concentrations of plastics in marine sediments from 42 coastal and estuarine sites spanning pollution gradients across south-eastern Australia. Acid digestion/density separation revealed 9552 individual microplastics from 2.84l of sediment across all samples; equating to a regional average of 3.4 microplastics·ml -1 sediment. Microplastics occurred as filaments (84% of total) and particle forms (16% of total). Positive correlations between microplastic filaments and wave exposure, and microplastic particles with finer sediments, indicate hydrological/sediment-matrix properties are important for deposition/retention. Contrary to expectations, positive relationships were not evident between microplastics and other pollutants (heavy metals/sewage), nor were negative relationships with neighbouring reef biota detected. Rather, microplastics were ubiquitous across sampling sites. Positive associations with some faunal-elements (i.e. invertebrate species richness) nevertheless suggest high potential for microplastic ingestion. Copyright © 2017 Elsevier Ltd. All rights reserved.

[Banana peel: a possible source of infection in the treatment of nipple fissures].

PubMed

Novak, Franz Reis; de Almeida, João Aprígio Guerra; de Souza e Silva, Rosana

2003-01-01

To study the microbiology of banana peel being sold in the city of Rio de Janeiro, in an attempt to determine the possibility that the peel may represent a source of infection for women who use it to treat nipple fissures. The following microorganisms were studied in 20 banana peel samples: mesophiles, total coliforms, fecal coliforms, Pseudomonas aeruginosa, lipolytic and proteolytic microorganisms, molds and yeasts, lactic bacteria, and coagulase-positive staphylococcus. The microbiological analyses revealed the occurrence of several typical groups of microorganisms, with the following distribution of positive results being detected in banana peel samples: mesophiles, 100%; total coliforms, 20%; coagulase-positive staphylococcus, 25%; molds and yeasts, 30%; proteolytic microorganisms, 70%; lipolytic microorganisms, 30%, and lactic bacteria, 95%. Fecal coliforms and Pseudomonas aeruginosa were not isolated. The results show the presence of potentially pathogenic microorganisms in levels which could compromise the microbiological quality of the banana peel. Its use for the treatment of nipple fissures can initiate an infectious process.

[The method and result analyses of pathogenic bacteria culture on chronic periprosthetic joint infection after total knee arthroplasty and total hip arthroplasty].

PubMed

Ji, Baochao; Xu, Enjie; Cao, Li; Yang, Desheng; Xu, Boyong; Guo, Wentao; Aili, Rehei

2015-02-01

To analyze the results of pathogenic bacteria culture on chronic periprosthetic joint infection after total knee arthroplasty (TKA) and total hip arthroplasty (THA). The medical data of 23 patients with chronic periprosthetic joint infection after TKA or THA from September 2010 to March 2014 were reviewed. Fifteen cases of TKA and 8 cases of THA were included in this study. There were 12 male and 11 female patients with the mean age of 62 years (range from 32 to 79 years), and among them 9 patients with sinus. All patients discontinued antibiotic therapy for a minimum of 2 weeks before arthrocentesis, taking pathogenic bacteria culture and antimicrobial susceptibility test by using synovial fluid taken preoperatively and intraoperatively of revision. Common pathogenic bacteria culture and pathological biopsy were taken on tissues intraoperatively of revision. Culture-negative specimens were prolonged the period of incubation for 2 weeks. The overall culture-positive rate of all 23 patients for 1 week before revision was 30.4% (7/23), and the positive rate of culture-negative samples which prolonged for 2 weeks was 39.1% (9/23). The overall culture-positive rate of patients for 1 week intraoperatively of revision was 60.9% (14/23), and the positive rate of culture-negative samples which prolonged for 2 weeks was 82.6% (19/23). The incubation results of 7 cases (30.4%) preoperatively conformed to that of intraoperation. The culture-positive rate of pathogenic bacteria culture can be increased evidently by discontinuing antimicrobial therapy for a minimum of 2 weeks prior to the definite diagnosis.

Use of the polymerase chain reaction to directly detect malaria parasites in blood samples from the Venezuelan Amazon.

PubMed

Laserson, K F; Petralanda, I; Hamlin, D M; Almera, R; Fuentes, M; Carrasquel, A; Barker, R H

1994-02-01

We have examined the reproducibility, sensitivity, and specificity of detecting Plasmodium falciparum using the polymerase chain reaction (PCR) and the species-specific probe pPF14 under field conditions in the Venezuelan Amazon. Up to eight samples were field collected from each of 48 consenting Amerindians presenting with symptoms of malaria. Sample processing and analysis was performed at the Centro Amazonico para la Investigacion y Control de Enfermedades Tropicales Simon Bolivar. A total of 229 samples from 48 patients were analyzed by PCR methods using four different P. falciparum-specific probes. One P. vivax-specific probe and by conventional microscopy. Samples in which results from PCR and microscopy differed were reanalyzed at a higher sensitivity by microscopy. Results suggest that microscopy-negative, PCR-positive samples are true positives, and that microscopy-positive and PCR-negative samples are true negatives. The sensitivity of the DNA probe/PCR method was 78% and its specificity was 97%. The positive predictive value of the PCR method was 88%, and the negative predictive value was 95%. Through the analysis of multiple blood samples from each individual, the DNA probe/PCR methodology was found to have an inherent reproducibility that was highly statistically significant.

Evaluation of Antibiotic Residues in Pasteurized and Raw Milk Distributed in the South of Khorasan-e Razavi Province, Iran.

PubMed

Moghadam, Mortez Mohammadzadeh; Amiri, Mostafa; Riabi, Hamed Ramezani Awal; Riabi, Hamid Ramezani Awal

2016-12-01

The presence of antibiotic residues in milk and other products livestock is a health problem which can endanger public health. Antibiotics are used widely in animal husbandry to treat diseases related to bacterial infections. Antimicrobial drugs have been in use for decades in industry. They are commonly used in livestock facilities to treat mastitis. This study aimed to investigate antibiotic residues in pasteurized milk distributed in schools, in milk collection centers, and in milk production factories in Gonabad city. This cross-sectional study was conducted on 251 samples of commercial pasteurized milk packet distributed in schools (code A), raw milk collection centers in Gonabad city (code B), and pasteurized milk production factories (code C) in Gonabad city. The Copan test kit of Denmark Christian Hansen Company was used to monitor antibiotic residues in milk. The data were analysed employing Chi-square test and one-way analysis of variance (ANOVA) to determine significant differences using SPSS software version 20. The significant level was considered at p<0.05. In total, 251 milk samples were collected out of which 143 (57%) were code A, 84 (33.5%) code B and 24 (9.6%) code C. Total number of 189 samples (75.2%) were negative and 62 (24.8%) were positive. From the three types of milk samples, 41 samples (28.7%) of the code A, 18 samples (21.4%) of the code B and 3 samples (12.5%) of the code C were positive. In general, from the milk samples most contaminated with antibiotics, 17 samples were positive in January and regarding code A, 13 samples were positive in the same month. There was not a significant difference among the three types of milk (p>0.05). The highest number of milk samples (n=7) contaminated with antibiotics were related to code B (38.5%). Most positive cases were related to code A in winter. Also, there was no significant difference among the three types of contaminated milk regarding the year and month (p=0.164 and p=0.917, respectively). Pasteurized milk supplied in the studied city has high level of contamination due to high use of antibiotics. A standard limit needs to be set for the right level of residue of antibiotics in milk to avoid the harmful effects.

Evaluation of Antibiotic Residues in Pasteurized and Raw Milk Distributed in the South of Khorasan-e Razavi Province, Iran

PubMed Central

Moghadam, Mortez Mohammadzadeh; Amiri, Mostafa; Riabi, Hamid Ramezani Awal

2016-01-01

Introduction The presence of antibiotic residues in milk and other products livestock is a health problem which can endanger public health. Antibiotics are used widely in animal husbandry to treat diseases related to bacterial infections. Antimicrobial drugs have been in use for decades in industry. They are commonly used in livestock facilities to treat mastitis. Aim This study aimed to investigate antibiotic residues in pasteurized milk distributed in schools, in milk collection centers, and in milk production factories in Gonabad city. Materials and Methods This cross-sectional study was conducted on 251 samples of commercial pasteurized milk packet distributed in schools (code A), raw milk collection centers in Gonabad city (code B), and pasteurized milk production factories (code C) in Gonabad city. The Copan test kit of Denmark Christian Hansen Company was used to monitor antibiotic residues in milk. The data were analysed employing Chi-square test and one-way analysis of variance (ANOVA) to determine significant differences using SPSS software version 20. The significant level was considered at p<0.05. Results In total, 251 milk samples were collected out of which 143 (57%) were code A, 84 (33.5%) code B and 24 (9.6%) code C. Total number of 189 samples (75.2%) were negative and 62 (24.8%) were positive. From the three types of milk samples, 41 samples (28.7%) of the code A, 18 samples (21.4%) of the code B and 3 samples (12.5%) of the code C were positive. In general, from the milk samples most contaminated with antibiotics, 17 samples were positive in January and regarding code A, 13 samples were positive in the same month. There was not a significant difference among the three types of milk (p>0.05). The highest number of milk samples (n=7) contaminated with antibiotics were related to code B (38.5%). Most positive cases were related to code A in winter. Also, there was no significant difference among the three types of contaminated milk regarding the year and month (p=0.164 and p=0.917, respectively). Conclusion Pasteurized milk supplied in the studied city has high level of contamination due to high use of antibiotics. A standard limit needs to be set for the right level of residue of antibiotics in milk to avoid the harmful effects. PMID:28208877

Development and validation of a Pneumocystis jirovecii real-time polymerase chain reaction assay for diagnosis of Pneumocystis pneumonia

PubMed Central

Church, Deirdre L; Ambasta, Anshula; Wilmer, Amanda; Williscroft, Holly; Ritchie, Gordon; Pillai, Dylan R; Champagne, Sylvie; Gregson, Daniel G

2015-01-01

BACKGROUND: Pneumocystis jirovecii (PJ), a pathogenic fungus, causes severe interstitial Pneumocystis pneumonia (PCP) among immunocompromised patients. A laboratory-developed real-time polyermase chain reaction (PCR) assay was validated for PJ detection to improve diagnosis of PCP. METHODS: Forty stored bronchoalveolar lavage (BAL) samples (20 known PJ positive [PJ+] and 20 known PJ negative [PJ−]) were initially tested using the molecular assay. Ninety-two sequentially collected BAL samples were then analyzed using an immunofluorescence assay (IFA) and secondarily tested using the PJ real-time PCR assay. Discrepant results were resolved by retesting BAL samples using another real-time PCR assay with a different target. PJ real-time PCR assay performance was compared with the existing gold standard (ie, IFA) and a modified gold standard, in which a true positive was defined as a sample that tested positive in two of three methods in a patient suspected to have PCP. RESULTS: Ninety of 132 (68%) BAL fluid samples were collected from immunocompromised patients. Thirteen of 92 (14%) BALs collected were PJ+ when tested using IFA. A total of 40 BAL samples were PJ+ in the present study including: all IFA positive samples (n=13); all referred PJ+ BAL samples (n=20); and seven additional BAL samples that were IFA negative, but positive using the modified gold standard. Compared with IFA, the PJ real-time PCR had sensitivity, specificity, and positive and negative predictive values of 100%, 91%, 65% and 100%, respectively. Compared with the modified gold standard, PJ real-time PCR had a sensitivity, specificity, and positive and negative predictive values of 100%. CONCLUSION: PJ real-time PCR improved detection of PJ in immunocompromised patients. PMID:26600815

Legionella species colonization in cooling towers: risk factors and assessment of control measures.

PubMed

Mouchtouri, Varvara A; Goutziana, Georgia; Kremastinou, Jenny; Hadjichristodoulou, Christos

2010-02-01

Cooling towers can be colonized by Legionella spp, and inhalation of aerosols generated by their operation may cause Legionnaires' disease in susceptible hosts. Environmental investigations of Legionnaires' disease outbreaks linked with cooling towers have revealed poorly maintained systems, lack of control measures, and failure of system equipment. The purpose of this study was to identify Legionella-contaminated cooling towers, identify risk factors for contamination, and assess the effectiveness of control measures. A total of 96 cooling towers of public buildings were registered and inspected, and 130 samples were collected and microbiologically tested. Microbiological test results were associated with characteristics of cooling towers, water samples, inspection results, and maintenance practices. Of the total 96 cooling towers examined, 47 (48.9%) were colonized by Legionella spp, and 22 (22.9%) required remedial action. A total of 65 samples (50.0%) were positive (> or = 500 cfu L(-1)), and 30 (23%) were heavily contaminated (> or = 10(4) cfu L(-1)). Of the 69 isolates identified, 55 strains (79.7.%) were L pneumophila. Legionella colonization was positively associated with the absence of training on Legionella control (relative risk [RR] = 1.66; P = .02), absence of regular Legionella testing (RR = 2.07: P = .002), absence of sunlight protection (RR = 1.63: P = .02), with samples in which the free residual chlorine level in the water sample was < 0.5 mg/L (RR = 2.23; P = .01), and with total plate count (P =.001). Colonization was negatively associated with chemical disinfection (RR = 0.2; P = .0003) and with the presence of a risk assessment and management plan (RR = 0.12; P = .0005). A statistically significant higher age (P =.01) was found in legionellae-positive cooling towers (median, 17 years; interquartile range [IQR] =5.0 to 26.0 years) compared with noncolonized cooling towers (median age, 6 years; IQR =1.0 to 13.5 years). After the 22 legionellae-positive cooling towers were disinfected with chlorine, 2 (9%) of them remained positive for Legionella spp with a concentration > or = 1000 cfu L(-1). Cooling towers can be heavily colonized by Legionella spp and thus present a potential risk for infection. This study demonstrates the importance of a risk assessment and management plan. Water chlorination effectively reduces legionellae contamination. Proper training of cooling tower operators is paramount. Copyright 2010 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

In-house PCR with DNA extracted directly from positive slides to confirm or exclude the diagnosis of tuberculosis: focus on biosafety.

PubMed

de Almeida, Isabela Neves; Aleixo, Agdemir Valéria; Carvalho, Wânia da Silva; de Miranda, Silvana Spindola

2015-01-01

The possibility to obtain DNA from smears is a valuable alternative to remedy the lack of samples when they are totally used for bacilloscopy; this technique solves the biosafety problem related to a possible accident with the transportation of flasks containing potentially transmissible clinical samples. Hence, the purpose of this study was to utilize the insertion sequence IS6110 for amplification of DNA from a smear-positive sample for tuberculosis (TB) diagnosis. Among the 52 positive bacilloscopies, sensitivity, specificity, positive predictive value and negative predictive value were 52.3%, 100%, 100% and 89.7%, respectively whereas accuracy was 90.7%. The IS6110-based PCR for TB diagnosis developed in DNA extracted from a positive smear is a fast, simple, specific, and safe method. Copyright © 2014 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Fluorometric determination of the DNA concentration in municipal drinking water.

PubMed Central

McCoy, W F; Olson, B H

1985-01-01

DNA concentrations in municipal drinking water samples were measured by fluorometry, using Hoechst 33258 fluorochrome. The concentration, extraction, and detection methods used were adapted from existing techniques. The method is reproducible, fast, accurate, and simple. The amounts of DNA per cell for five different bacterial isolates obtained from drinking water samples were determined by measuring DNA concentration and total cell concentration (acridine orange epifluorescence direct cell counting) in stationary pure cultures. The relationship between DNA concentration and epifluorescence total direct cell concentration in 11 different drinking water samples was linear and positive; the amounts of DNA per cell in these samples did not differ significantly from the amounts in pure culture isolates. We found significant linear correlations between DNA concentration and colony-forming unit concentration, as well as between epifluorescence direct cell counts and colony-forming unit concentration. DNA concentration measurements of municipal drinking water samples appear to monitor changes in bacteriological quality at least as well as total heterotrophic plate counting and epifluorescence direct cell counting. PMID:3890737

Tularaemia seroprevalence of captured and wild animals in Germany: the fox (Vulpes vulpes) as a biological indicator.

PubMed

Kuehn, A; Schulze, C; Kutzer, P; Probst, C; Hlinak, A; Ochs, A; Grunow, R

2013-04-01

A total of 2475 animals from Germany, both captive and wild, were tested for antibodies against Francisella tularensis to obtain more knowledge about the presence of this pathogen in Germany. An indirect and a competitive ELISA served as screening methods, positive and inconclusive samples were confirmed by Western blot. Of the zoo animals sampled between 1992 and 2007 (n = 1122), three (0·3%) were seropositive. The seroconversion of a hippopotamus in Berlin Zoo was documented. From 1353 serum samples of wild foxes (Vulpes vulpes), raccoon dogs (Nyctereutes procyonoides) and wild boars (Sus scrofa), collected between 2005 and 2009 in the federal state of Brandenburg (surrounding Berlin), a total of 101 (7·5%) tested positive for antibodies to F. tularensis lipopolysaccharide. Our results indicate a higher seroprevalence of F. tularensis in wildlife in eastern Germany than commonly assumed. Furthermore, we found foxes and raccoon dogs to be biological indicators for tularaemia.

Avian Influenza Virus (H11N9) in Migratory Shorebirds Wintering in the Amazon Region, Brazil

PubMed Central

de Araujo, Jansen; de Azevedo Júnior, Severino M.; Gaidet, Nicolas; Hurtado, Renata F.; Walker, David; Thomazelli, Luciano M.; Ometto, Tatiana; Seixas, Marina M. M.; Rodrigues, Roberta; Galindo, Daniele B.; da Silva, Adriana C. S.; Rodrigues, Arlinéa M. M.; Bomfim, Leonardo L.; Mota, Marcelo A.; Larrazábal, Maria E.; Branco, Joaquim O.; Serafini, Patricia; Neto, Isaac S.; Franks, John; Webby, Richard J.; Webster, Robert G.; Durigon, Edison L.

2014-01-01

Aquatic birds are the natural reservoir for avian influenza viruses (AIV). Habitats in Brazil provide stopover and wintering sites for water birds that migrate between North and South America. The current study was conducted to elucidate the possibility of the transport of influenza A viruses by birds that migrate annually between the Northern and Southern Hemispheres. In total, 556 orotracheal/cloacal swab samples were collected for influenza A virus screening using real-time RT-PCR (rRT-PCR). The influenza A virus-positive samples were subjected to viral isolation. Four samples were positive for the influenza A matrix gene by rRT-PCR. From these samples, three viruses were isolated, sequenced and characterized. All positive samples originated from a single bird species, the ruddy turnstone (Arenaria interpres), that was caught in the Amazon region at Caeté Bay, Northeast Pará, at Ilha de Canelas. To our knowledge, this is the first isolation of H11N9 in the ruddy turnstone in South America. PMID:25329399

Significance of serum antibody test for toxocariasis in healthy healthcare examinees with eosinophilia in Seoul and Gyeongsangnam-do, Korea.

PubMed

Kim, Hong Seok; Jin, Yan; Choi, Min-Ho; Kim, Jae-Hwan; Lee, Young Ha; Yoon, Cheong Ha; Hwang, Eui-Hyuk; Kang, Hun; Ahn, Sang-Yong; Kim, Gi Jin; Hong, Sung-Tae

2014-12-01

There have been numerous reports on the relationship between eosinophilia and toxocariasis. The present study investigated seropositive rates of toxocariasis among healthy people with or without eosinophilia in urban and rural areas, and assessed risk factors for positive antibody test. A total of 610 healthy people, who visited health check-up (Medicheck®, Korea Association of Health Promotion), 310 from Seoul and 300 from Gyeongsangnam-do, were subjected for this study. Their serum samples were tested by ELISA with the crude antigen of Toxocara canis larvae. Cross-reactions with other tissue invading helminth antigens were also investigated. Total antibody positive rate of toxocariasis was 8.7% of the 610 subjects. When the subjects were grouped into 3 by their eosinophil counts, the antibody positive rates significantly differed by the groups; 5.9% (18/306) in the group<350/µL, 10.0% (11/110) in the group 350-500/µL, and 12.4% (24/194) in the group>500/µL (P=0.028). A total of 22 serum samples cross-reacted with other tissue-invading helminth antigens. A questionnaire analysis recognized drinking alcohol and smoking as significant risk factors of toxocariasis. In conclusion, toxocariasis antibody positive rate is correlated with eosinophil counts. It is recommended that healthy subjects with eosinophilia by routine health examination and risk factors undergo Toxocara serology by multiantigen ELISA to investigate etiology.

Sleep duration and its association with demographics, lifestyle factors, poor mental health and chronic diseases in older Chinese adults.

PubMed

Wang, Shibin; Wu, Yanhua; Ungvari, Gabor S; Ng, Chee H; Forester, Brent P; Gatchel, Jennifer R; Chiu, Helen F K; Kou, Changgui; Fu, Yingli; Qi, Yue; Yu, Yaqin; Li, Bo; Xiang, Yu-Tao

2017-11-01

This study investigated the total sleep time (TST) and its associated factors in an older Chinese adult population. Multistage stratified cluster sampling was used in this cross-sectional study. A total of 4,115 older adults aged 60 to 79 years were selected and interviewed. Sleep duration was classified as short (<7h per day), long (>8h per day) and medium sleep (7-8h per day). The total mean sleep time was 6.86±1.75h. Short and long sleepers accounted for 45.2% and 14.8% of the sample, respectively. Multivariate logistic regression analysis revealed that inadequate fruit intake and poor mental health were positively associated with short sleep, and married/cohabitation status and living in rural areas was negatively associated with short sleep. In addition, aged 75-79 years old, inadequate fruit intake, poor mental health and multi-morbidity were positively associated with long sleep. Ischemic heart disease, COPD and chronic gastroenteritis/peptic ulcer were positively associated with short sleep duration, while hyperlipidemia, hypertension, cerebrovascular diseases, and urolithiasis were positively associated with long sleep duration. Given the high frequency of aberrant sleep duration and its negative health impact, health professionals should pay more attention to sleep patterns in older people. Copyright © 2017 Elsevier B.V. All rights reserved.

Comparison of fecal pooling strategies for detection of Mycobacterium avium ssp. paratuberculosis in cattle.

PubMed

McKenna, S L B; Ritter, C; Dohoo, I; Keefe, G P; Barkema, H W

2018-05-23

In herds with typical moderate to low within-herd prevalence, testing for Mycobacterium avium ssp. paratuberculosis (MAP), the infectious agent of Johne's disease, will be more cost-effective if individual fecal samples are cultured in composite pools. However, sensitivity to classify a pool containing 1 or more positive individual samples as positive may depend on pool size and number of individual positive samples within a pool. Fecal samples collected from 994 dairy cows sampled at slaughter were cultured to detect MAP. Culturing was done both individually and as composite pooled samples using the TREK ESP Culture System II broth medium (Thermo Fisher Scientific, Trek Diagnostic Systems Inc., Cleveland, OH). Composite samples consisted of pools containing feces from 3, 5, 8, 10, or 15 cows. The number of individual fecal culture-positive cows within each pool ranged from 0 to 4. Culture of individual fecal samples detected MAP in 36 (3.6%) of the 994 cows. Individual samples that were detected within the first 50 d by TREK ESP Culture System II were more likely to lead to a positive pool result. In total, 840 pooled fecal samples were examined for presence of MAP, and of those, 272 pools actually contained feces from fecal culture-positive cows. The crude sensitivity (proportion of pools that contained at least 1 fecal-positive cow that tested positive) for pools of 3, 5, 8, 10, and 15 was 47, 67, 44, 59, and 39%, respectively. Across pools, an increase of the number of fecal culture-positive samples from 1 to 2 enhanced overall crude sensitivity from 44 to 71%. However, sensitivity did not further increase for pools with 3 or 4 fecal culture-positive samples (63 and 60%, respectively). Additionally, a simulation analysis assessing probability of pooled fecal samples being positive in herds of 50 and 100 cows was conducted. The simulation assumed that 1, 2, or 5 cows per herd were MAP fecal culture-positive and that pools of 5 and 10 were used. This low-prevalence herd simulation indicated that weighted mean herd probabilities of detecting a positive herd ranged between 52 and 99.3%, with the lowest probability for pools of 10 with 1 positive cow in the herd and the highest probability for pools of 5 with 5 positive cows in the herd. However, overall, pools of 5 and 10 had similar diagnostic capabilities, enabling cost savings by utilizing pools of 10. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Working group session report: Neutron beam line shielding.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Russell, G. J.; Ikedo, Y.

2001-01-01

We have examined the differences between a 2-D model and a 3-D model for designing the beam-line shield for the HIPPO instrument at the Lujan Center at the Los Alamos National Laboratory. We have calculated the total (neutron and gamma ray) dose equivalent rate coming out of the personal access ports from the HIPPO instrument experiment cave. In order to answer this question, we have investigated two possible worst-case scenarios: (a) failure of the T{sub 0}-chopper and no sample at the sample position; and (b) failure of the T{sub 0}-chopper with a thick sample (a piece of Inconel-718, 10 cmmore » diam by 30 cm long) at the sample position.« less

[Contamination of human milk with aerobic flora: Evaluation of losses for a human milk bank].

PubMed

Dewitte, C; Courdent, P; Charlet, C; Dumoulin, D; Courcol, R; Pierrat, V

2015-05-01

In France, human milk banks pasteurize milk for the mother's own hospitalized baby (personalized milk) and for donation. There is specific legislation regulating the activity of human milk banks with bacterial screening of donor milk before and after pasteurization. Milk should be tested for Staphylococcus aureus and total aerobic flora. Any sample of milk positive for aerobic flora and/or S. aureus before and/or after pasteurization should be discarded. The real pathogenicity of the total aerobic flora is actually debated as well as the usefulness of systematic postpasteurization screening. The aim of this study was to quantify milk losses related to prepasteurization contamination by total aerobic flora in a regional milk bank, to identify losses due to contamination with S. aureus or aerobic flora, and to analyze differences between centers. This was a prospective observational study conducted in the regional human milk bank of the Nord-Pas-de-Calais area in France. Data were collected from six major centers providing 80% of the milk collected between June 2011 and June 2012. Variables were the volumes of personalized milk collected by each center, volumes of contaminated milk, and the type of bacteria identified. During the study period, the regional human milk bank treated 4715 L (liters) of personalized milk and 508 L (10.8%) were discarded due to bacteriological screening. Among these 508 L, 43% were discarded because of a prepasteurization contamination with aerobic flora, 55% because of a prepasteurization contamination with S. aureus, and 2% because of other pathogenic bacteria. Postpasteurization tests were positive in 25 samples (0.5%). Only five of these 25 samples were positive before pasteurization and in all cases with S. aureus. A total of 218 L were destroyed because of prepasteurization contamination with total aerobic flora, while the postpasteurization culture was sterile. There was a great difference between centers in the percentage of discarded milk and the type of contamination. The percentage of discarded milk varied from 4 to 16% (P<0.001) and the percentage of prepasteurization positive samples with aerobic flora from 0 to 70% (P<0.001). Costing 80 €/L in France, this represented an economic loss of €17,440. A significant volume of milk is discarded because of contamination with total aerobic flora found only in prepasteurization tests. Reassessment of the French regulations with regard to microbiological safety could save human milk to cover the needs of a larger group of preterm babies. Copyright © 2015. Published by Elsevier SAS.

Campylobacter spp., Giardia spp., Cryptosporidium spp., Noroviruses, and Indicator Organisms in Surface Water in Southwestern Finland, 2000-2001

PubMed Central

Hörman, Ari; Rimhanen-Finne, Ruska; Maunula, Leena; von Bonsdorff, Carl-Henrik; Torvela, Niina; Heikinheimo, Annamari; Hänninen, Marja-Liisa

2004-01-01

A total of 139 surface water samples from seven lakes and 15 rivers in southwestern Finland were analyzed during five consecutive seasons from autumn 2000 to autumn 2001 for the presence of various enteropathogens (Campylobacter spp., Giardia spp., Cryptosporidium spp., and noroviruses) and fecal indicators (thermotolerant coliforms, Escherichia coli, Clostridium perfringens, and F-RNA bacteriophages) and for physicochemical parameters (turbidity and temperature); this was the first such systematic study. Altogether, 41.0% (57 of 139) of the samples were positive for at least one of the pathogens; 17.3% were positive for Campylobacter spp. (45.8% of the positive samples contained Campylobacter jejuni, 25.0% contained Campylobacter lari, 4.2% contained Campylobacter coli, and 25.0% contained Campylobacter isolates that were not identified), 13.7% were positive for Giardia spp., 10.1% were positive for Cryptosporidium spp., and 9.4% were positive for noroviruses (23.0% of the positive samples contained genogroup I and 77.0% contained genogroup II). The samples were positive for enteropathogens significantly (P < 0.05) less frequently during the winter season than during the other sampling seasons. No significant differences in the prevalence of enteropathogens were found when rivers and lakes were compared. The presence of thermotolerant coliforms, E. coli, and C. perfringens had significant bivariate nonparametric Spearman's rank order correlation coefficients (P < 0.001) with samples that were positive for one or more of the pathogens analyzed. The absence of these indicators in a logistic regression model was found to have significant predictive value (odds ratios, 1.15 × 108, 7.57, and 2.74, respectively; P < 0.05) for a sample that was negative for the pathogens analyzed. There were no significant correlations between counts or count levels for thermotolerant coliforms or E. coli or the presence of F-RNA phages and pathogens in the samples analyzed. PMID:14711629

Peste des petits ruminants infection in domestic ruminants in Sudan.

PubMed

Intisar, K S; Ali, Y H; Haj, M A; Sahar, M A T; Shaza, M M; Baraa, A M; Ishag, O M; Nouri, Y M; Taha, K M; Nada, E M; Ahmed, A M; Khalafalla, A I; Libeau, G; Diallo, A

2017-04-01

The existence of peste des petits ruminants (PPR) in domestic ruminants and camels in Sudan during 2008-2012 was investigated. Lung tissues and serum samples were randomly collected from sheep, goats, cattle, and camels at different areas of Sudan. A total of 12,384 serum samples were collected from clinically healthy 7413 sheep, 1988 camels, 1501 cattle, 1459 goats, and 23 gazelles at different areas in the Sudan. They were examined for PPR antibodies using competitive ELISA (cELISA). The overall detected seroprevalence of PPR in tested sera was 49.4%; seroprevalence values within species were 67.1, 48.2, 25.8, 2.1, and 21.7% in sheep, goat, cattle, camels, and gazelles, respectively. The highest seroprevalence (68.1%) was observed in sera collected from Darfur states, then the central states (54.3%). A total of 1276 lung tissue samples (623 sheep, 324 cattle, 220 camels, and 109 goats) were collected. The majority of lung samples were collected from clinically healthy animals that showed lesions on PM in slaughterhouses (95%) and during PPR outbreaks; samples were tested for PPR antigen using immunocapture ELISA (IcELISA). PPR antigen was detected in 233 out of the 1276 tested samples (18.3%). Positive results were observed in samples collected from clinically healthy and diseased animals. The observed prevalence values in each species were 33.6, 21.1, 15.4, and 12.3% in camel, goat, sheep, and cattle, respectively. PPR antigen was detected in samples from different areas; however, the highest prevalence (63.9%) was found in samples collected from the eastern states, then Khartoum state (28%). Trials for virus isolation were done in different cell cultures. Out of 30 IcELISA-positive samples inoculated in primary bovine and ovine kidney cells, Vero cells, the PPR virus was successfully isolated from 15 (eight sheep, five camels, and two goats) samples in the three cell culture types. Using RT-PCR, PPRV nucleic acid was detected in all 25 IcELISA-positive tested samples.

Microbiological evaluation of water during the 2011 flood crisis in Thailand.

PubMed

Chaturongkasumrit, Yuphakhun; Techaruvichit, Punnida; Takahashi, Hajime; Kimura, Bon; Keeratipibul, Suwimon

2013-10-01

In 2011, a severe flood occurred in Thailand, covering nearly half the country in water for several months. The contamination of floodwater and subsequent contamination of water for human consumption could have potentially led to a widespread health crisis. However, to date, no study has been conducted to determine the safety of the waters used for human consumption in Thailand during the severe flood. Therefore, we conducted microbiological analysis of 4 kinds of water (floodwater, river water, tap water, and filtered tap water) collected from industrial and residential areas that were damaged due to flooding. Higher net levels of bacteria were found in water with a higher turbidity. No clear trend was observed in the pH value of all 4 water samples. The level of total bacterial contamination in the water samples was estimated by real-time quantitative polymerase chain reaction (PCR). Eleven of the 12 tap water samples and all of the filtered tap water samples had a total bacterial load that exceeded the Thai water quality standards. One of the tap water samples and one of the filtered tap water samples were found to be positive for Shigella sp., although none of the floodwater samples showed detectable levels of this pathogen as determined by PCR analysis. One of the samples of floodwater was also found to be positive for Leptospira sp., but none of the tap water or filtered tap water samples were positive. Most of the tap water samples and all filtered tap water samples were found to be contaminated with Vibrio cholerae. Bacterial contamination in water samples was also analyzed by denaturing gradient gel electrophoresis (DGGE) analysis. These results revealed that several microorganisms were transferred via floodwater to different areas in the central part of Thailand and cross-contaminated between floodwater and water for human consumption. © 2013.

Intrafamilial clustering of anti-ATLA-positive persons.

PubMed

Kajiyama, W; Kashiwagi, S; Hayashi, J; Nomura, H; Ikematsu, H; Okochi, K

1986-11-01

A total of 1,333 persons in 627 families were surveyed for presence of antibody to adult T-cell leukemia-associated antigen (anti-ATLA). Each person was classified according to the anti-ATLA status (positive for sample 1, negative for sample 2) of the head of household of his or her family. In sample 1, the sex- and age-standardized prevalence of anti-ATLA was 38.5%. This was five times as high as the standardized prevalence in sample 2 (7.8%). There were significant differences in prevalence of anti-ATLA between males in samples 1 and 2 and between females in samples 1 and 2. In every age group, prevalence in sample 1 was greater than that in sample 2 except for males aged 60-69 years. In each of four subareas, families in sample 1 had higher standardized prevalence (29.6-42.5%) than families in sample 2 (6.0-9.7%). Although crude prevalence decreased with family size in sample 1 (62.1-25.4%) as well as in sample 2, indirectly standardized prevalence was almost equal within each sample, regardless of number of family members. The degree of aggregation was independent of locality and family size. These data suggest that anti-ATLA-positive persons aggregate in family units.

Performance Evaluation of the Elecsys Syphilis Assay for the Detection of Total Antibodies to Treponema pallidum

PubMed Central

Enders, Martin; Hunjet, Andrea; Gleich, Michael; Imdahl, Roland; Mühlbacher, Annelies; Schennach, Harald; Chaiwong, Kriangsak; Sakuldamrongpanich, Tasanee; Turhan, Ajda; Sertöz, Rüchan; Wolf, Eva; Mayer, Wolfgang; Tao, Chuanmin; Wang, Lan Lan; Semprini, Simona

2014-01-01

Syphilis is a health problem of increasing incidence in recent years that may have severe complications if not diagnosed and treated at an early stage. There are many diagnostic tests available for syphilis, but there is no gold standard, and diagnosis therefore usually relies upon a combination of tests. In this multicenter study, we evaluated the treponemal Elecsys syphilis assay for use in the diagnosis of syphilis in routine samples, i.e., when syphilis is suspected or during antenatal or blood donation screening. The sensitivity and specificity of the Elecsys syphilis assay were compared head to head with those of other treponemal assays used in routine clinical practice and were assessed in potentially cross-reactive samples from patients with Epstein-Barr virus, HIV, and Lyme disease. In a total of 8,063 syphilis-negative samples collected from routine diagnostic requests and blood donations, the Elecsys syphilis assay had a specificity of 99.88%. In 928 samples previously identified as syphilis positive, the sensitivity was 99.57 to 100% (the result is presented as a range depending on whether four initially indeterminate samples are included in the assessment). The specificity of the Elecsys syphilis assay in patients with other infections was 100%; no false-positive samples were identified. PMID:25355799

Associations between emotion regulation difficulties, eating disorder symptoms, non-suicidal self-injury, and suicide attempts in a heterogeneous eating disorder sample.

PubMed

Pisetsky, Emily M; Haynos, Ann F; Lavender, Jason M; Crow, Scott J; Peterson, Carol B

2017-02-01

This study examined the associations between specific dimensions of emotion dysregulation and eating disorder (ED) symptoms and behaviors, non-suicidal self-injury (NSSI), and suicide attempts in a heterogeneous ED sample. Participants (N=110) completed the Difficulties in Emotion Regulation Scale (DERS), the Eating Disorder Examination Questionnaire (EDE-Q), and self-reported the presence of lifetime NSSI and a lifetime suicide attempt. The EDE-Q global score, a primarily cognitive measure of ED symptoms, was significantly positively correlated with DERS strategies, clarity, and awareness subscale scores and DERS total score (ps<0.01). Only the strategies subscale was uniquely positively associated with EDE-Q global score in a multivariate regression analysis. There was no association between the frequency of binge eating or frequency of driven exercise and any of the DERS subscale scores or total score (ps>0.01). Frequency of purging was significantly, positively associated with DERS impulse subscale score and total score (p<0.01). None of the DERS subscale scores were significantly different between those with and without NSSI or between those with and without a lifetime suicide attempt (ps>0.01). Findings indicate that in a heterogeneous ED sample, emotion regulation deficits are more strongly associated with cognitively-oriented symptoms of EDs than behavioral symptoms such as a binge eating, purging, driven exercise, NSSI, or suicide attempts. Copyright © 2016 Elsevier Inc. All rights reserved.

Evaluation of the Architect HIV Ag/Ab Combo Assay in a low-prevalence setting: The role of samples with a low S/CO ratio.

PubMed

Alonso, Roberto; Pérez-García, Felipe; Gijón, Paloma; Collazos, Ana; Bouza, Emilio

2018-06-01

The Architect HIV Ag/Ab Combo Assay, a fourth-generation ELISA, has proven to be highly reliable for the diagnosis of HIV infection. However, its high sensitivity may lead to false-positive results. To evaluate the diagnostic performance of Architect in a low-prevalence population and to assess the role of the sample-to-cutoff ratio (S/CO) in reducing the frequency of false-positive results. We conducted a retrospective study of samples analyzed by Architect between January 2015 and June 2017. Positive samples were confirmed by immunoblot (RIBA) or nucleic acid amplification tests (NAATs). Different S/CO thresholds (1, 2.5, 10, 25, and 100) were analyzed to determine sensitivity, specificity, and negative and positive predictive values (NPV, PPV). ROC analysis was used to determine the optimal S/CO. A total of 69,471 samples were analyzed. 709 (1.02%) were positive by Architect. Of these, 63 (8.89%) were false-positive results. Most of them (93.65%) were in samples with S/CO < 100. However, most confirmations by NAATs (12 out of 19 cases) were also recorded for these samples. The optimal S/CO was 2.5, which provided the highest area under the ROC curve (0.9998) and no false-negative results. With this S/CO, sensitivity and specificity were 100.0%, and PPV and NPV were 95.8% and 100.0%, respectively. In addition, the frequency of false-positive results decreased significantly to 4.15%. Although Architect generates a relatively high number of false-positive results, raising the S/CO limit too much to increase specificity can lead to false-negative results, especially in newly infected individuals. Copyright © 2018 Elsevier B.V. All rights reserved.

Direct detection of Trichomonas vaginalis virus in Trichomonas vaginalis positive clinical samples from the Netherlands.

PubMed

Jehee, Ivo; van der Veer, Charlotte; Himschoot, Michelle; Hermans, Mirjam; Bruisten, Sylvia

2017-12-01

Trichomonas vaginalis is the most common sexually transmitted parasitical infection worldwide. T. vaginalis can carry a virus: Trichomonas vaginalis virus (TVV). To date, four TVV species have been described. Few studies have investigated TVV prevalence and its clinical importance. We have developed a nested reverse-transcriptase PCR, with novel, type specific primers to directly detect TVV RNA in T. vaginalis positive clinical samples. A total of 119T. vaginalis positive clinical samples were collected in Amsterdam and "s-Hertogenbosch, the Netherlands, from 2012 to 2016. For all samples T. vaginalis was genotyped using multi-locus sequence typing. The T. vaginalis positive samples segregated into a two-genotype population: type I (n=64) and type II (n=55). All were tested for TVV with the new TVV PCR. We detected 3 of the 4 TVV species. Sequencing of the amplified products showed high homology with published TVV genomes (82-100%). Half of the T. vaginalis clinical samples (n=60, 50.4%) were infected with one or more TVV species, with a preponderance for TVV infections in T. vaginalis type I (n=44, 73.3%). Clinical data was available for a subset of samples (n=34) and we observed an association between testing positive for (any) TVV and reporting urogenital symptoms (p=0.023). The nested RT-PCR allowed for direct detection of TVV in T. vaginalis positive clinical samples. This may be helpful in studies and clinical settings, since T. vaginalis disease and/or treatment outcome may be influenced by the protozoa"s virus. Copyright © 2017 Elsevier B.V. All rights reserved.

Ammonia-Oxidizing Archaea Show More Distinct Biogeographic Distribution Patterns than Ammonia-Oxidizing Bacteria across the Black Soil Zone of Northeast China.

PubMed

Liu, Junjie; Yu, Zhenhua; Yao, Qin; Sui, Yueyu; Shi, Yu; Chu, Haiyan; Tang, Caixian; Franks, Ashley E; Jin, Jian; Liu, Xiaobing; Wang, Guanghua

2018-01-01

Black soils (Mollisols) of northeast China are highly productive and agriculturally important for food production. Ammonia-oxidizing microbes play an important role in N cycling in the black soils. However, the information related to the composition and distribution of ammonia-oxidizing microbes in the black soils has not yet been addressed. In this study, we used the amoA gene to quantify the abundance and community composition of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) across the black soil zone. The amoA abundance of AOA was remarkably larger than that of AOB, with ratios of AOA/AOB in the range from 3.1 to 91.0 across all soil samples. The abundance of AOA amoA was positively correlated with total soil C content ( p < 0.001) but not with soil pH ( p > 0.05). In contrast, the abundance of AOB amoA positively correlated with soil pH ( p = 0.009) but not with total soil C. Alpha diversity of AOA did not correlate with any soil parameter, however, alpha diversity of AOB was affected by multiple soil factors, such as soil pH, total P, N, and C, available K content, and soil water content. Canonical correspondence analysis indicated that the AOA community was mainly affected by the sampling latitude, followed by soil pH, total P and C; while the AOB community was mainly determined by soil pH, as well as total P, C and N, water content, and sampling latitude, which highlighted that the AOA community was more geographically distributed in the black soil zone of northeast China than AOB community. In addition, the pairwise analyses showed that the potential nitrification rate (PNR) was not correlated with alpha diversity but weakly positively with the abundance of the AOA community ( p = 0.048), whereas PNR significantly correlated positively with the richness ( p = 0.003), diversity ( p = 0.001) and abundance ( p < 0.001) of the AOB community, which suggested that AOB community might make a greater contribution to nitrification than AOA community in the black soils when ammonium is readily available.

Ammonia-Oxidizing Archaea Show More Distinct Biogeographic Distribution Patterns than Ammonia-Oxidizing Bacteria across the Black Soil Zone of Northeast China

PubMed Central

Liu, Junjie; Yu, Zhenhua; Yao, Qin; Sui, Yueyu; Shi, Yu; Chu, Haiyan; Tang, Caixian; Franks, Ashley E.; Jin, Jian; Liu, Xiaobing; Wang, Guanghua

2018-01-01

Black soils (Mollisols) of northeast China are highly productive and agriculturally important for food production. Ammonia-oxidizing microbes play an important role in N cycling in the black soils. However, the information related to the composition and distribution of ammonia-oxidizing microbes in the black soils has not yet been addressed. In this study, we used the amoA gene to quantify the abundance and community composition of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) across the black soil zone. The amoA abundance of AOA was remarkably larger than that of AOB, with ratios of AOA/AOB in the range from 3.1 to 91.0 across all soil samples. The abundance of AOA amoA was positively correlated with total soil C content (p < 0.001) but not with soil pH (p > 0.05). In contrast, the abundance of AOB amoA positively correlated with soil pH (p = 0.009) but not with total soil C. Alpha diversity of AOA did not correlate with any soil parameter, however, alpha diversity of AOB was affected by multiple soil factors, such as soil pH, total P, N, and C, available K content, and soil water content. Canonical correspondence analysis indicated that the AOA community was mainly affected by the sampling latitude, followed by soil pH, total P and C; while the AOB community was mainly determined by soil pH, as well as total P, C and N, water content, and sampling latitude, which highlighted that the AOA community was more geographically distributed in the black soil zone of northeast China than AOB community. In addition, the pairwise analyses showed that the potential nitrification rate (PNR) was not correlated with alpha diversity but weakly positively with the abundance of the AOA community (p = 0.048), whereas PNR significantly correlated positively with the richness (p = 0.003), diversity (p = 0.001) and abundance (p < 0.001) of the AOB community, which suggested that AOB community might make a greater contribution to nitrification than AOA community in the black soils when ammonium is readily available. PMID:29497404

Detection and molecular characterization of Cryptosporidium spp. in captive canaries (Serinus canaria) using different diagnostic methods.

PubMed

Camargo, Vinícius da Silva; Santana, Bruna Nicoleti; Ferrari, Elis Domingos; Nakamura, Alex Akira; Nagata, Walter Bertequini; Nardi, Ana Rita Moraes; Meireles, Marcelo Vasconcelos

2018-01-01

This study used several diagnostic methods to examine the occurrence of and molecularly characterize Cryptosporidium spp. in captive canaries (Serinus canaria) in southern and southeastern Brazil. A total of 498 fecal samples were purified by centrifugal-flotation using Sheather's solution. Cryptosporidium spp. diagnosis was performed using three diagnostic methods: malachite green negative staining, nested PCR targeting the 18S rRNA gene, followed by sequencing the amplified fragments, and duplex real-time PCR targeting the 18S rRNA specific to detect Cryptosporidium galli and Cryptosporidium avian genotype III. The overall positivity for Cryptosporidium spp. (total samples positive in at least one protocol) from the microscopic analysis, nested PCR and duplex real-time PCR protocol results was 13.3% (66/498). The positivity rates were 2.0% (10/498) and 4.6% (23/498) for Cryptosporidium spp. by microscopy and nested PCR, respectively. Sequencing of 20 samples amplified by nested PCR identified C. galli (3.0%; 15/498), Cryptosporidium avian genotype I (0.8%; 4/498) and Cryptosporidium avium (0.2%; 1/498). Duplex real-time PCR revealed a positivity of 7.8% (39/498) for C. galli and 2.4% (12/498) for avian genotype III. Malachite green negative staining differed significantly from nested PCR in detecting Cryptosporidium spp. Duplex real-time PCR was more sensitive than nested PCR/sequencing for detecting gastric Cryptosporidium in canaries.

Comparative performance of novel self-sampling methods in detecting high-risk human papillomavirus in 30,130 women not attending cervical screening.

PubMed

Bosgraaf, Remko P; Verhoef, Viola M J; Massuger, Leon F A G; Siebers, Albert G; Bulten, Johan; de Kuyper-de Ridder, Gabriëlle M; Meijer, Chris J M; Snijders, Peter J F; Heideman, Daniëlle A M; IntHout, Joanna; van Kemenade, Folkert J; Melchers, Willem J G; Bekkers, Ruud L M

2015-02-01

We determined whether the participation rate for a brush-based cervicovaginal self-sampling device is noninferior to the participation rate for a lavage-based one for testing for hrHPV (high-risk human papillomavirus). Additionally, positivity rates for hrHPV, the detection rates for cervical intraepithelial neoplasia grades 2 and 3 or worse (CIN2+/3+), and user comfort were compared. A total of 35,477 non-responders of the regular cervical screening program aged 33-63 years were invited to participate. Eligible women (n = 30,130) were randomly assigned to receive either a brush-based or a lavage-based device, and a questionnaire for reporting user convenience. Self-sampling responders testing hrHPV-positive were invited for a physician-taken sample for cytology; triage-positive women were referred for colposcopy. A total of 5,218 women participated in the brush-based sampling group (34.6%) and 4809 women in the lavage-based group (31.9%), i.e. an absolute difference of 2.7% (95%CI 1.8-4.2). The hrHPV-positivity rates in the two groups were identical (8.3%, relative risk (RR) 0.99, 95%CI 0.87-1.13). The detection of CIN2+ and CIN3+ in the brush group (2.0% for CIN2+; 1.3% for CIN3+) was similar to that in the lavage group (1.9% for CIN2+; 1.0% for CIN3+) with a cumulative RR of 1.01, 95%CI 0.83-1.24 for CIN2+ and 1.25, 95%CI 0.92-1.70 for CIN3+. The two self-sampling devices performed similarly in user comfort. In conclusion, offering a brush-based device to non-responders is noninferior to offering a lavage-based device in terms of participation. The two self-sampling methods are equally effective in detecting hrHPV, CIN2+/CIN3+ and are both well accepted. © 2014 UICC.

Waterborne parasites: a current status from the Philippines.

PubMed

Onichandran, Subashini; Kumar, Thulasi; Salibay, Cristina C; Dungca, Julieta Z; Tabo, Hazel Al; Tabo, Norbel; Tan, Tian-Chye; Lim, Yvonne Al; Sawangjaroen, Nongyao; Phiriyasamith, Sucheep; Andiappan, Hemah; Ithoi, Init; Lau, Yee-Ling; Nissapatorn, Veeranoot

2014-05-28

Despite the amount of awareness created, waterborne disease still poses threat, especially in developing countries. Due to the scarcity of reported data on waterborne parasites, the consumption of unsafe water prolongs. Thus, the occurrences of waterborne parasites from various samples were investigated from one of the Southeast Asian country, the Philippines. A total of thirty three samples, each consisting of twelve liters, were collected and processed to obtain the sediment. Ten liters of sample each was processed to detect Cryptosporidium spp. and Giardia spp. using an immunomagnetic separation method prior to enumeration via fluorescence microscope. Meanwhile, the remaining two liters were cultured to detect Acanthamoeba and Naegleria through microscopy examination and polymerase chain reaction (PCR) analysis. Twelve samples (36.4%) from river (5), swimming pool (1), pond (3), rain tank (1), and natural lake (2) were positive for Cryptosporidium spp., 17 (45.5%) samples from river (9), pond (2), swimming pool (1), rain tank (1), and natural lake (4) were positive for Giardia spp. while, 13 (33.3%) samples from river (3), swimming pool (2), pond (2), dispenser (1), well (1), tap (2) and natural lake (2) were positive for Acanthamoeba spp. and 5 (18.2%) samples from river (1), natural lake (1), tap (1), dispenser (1) and mineral (1) were Naegleria spp. positive. Physical parameters such as temperature, conductivity, total dissolved solid (TDS), salinity, dissolved oxygen (DO), pH, and turbidity and chemical parameters such as ammonia, chlorine, fluoride, nitrate and nitrite were also measured. The highest chemical contamination was observed at pond 2. A good correlation was observed between Giardia and nitrite (r = 0.736, p < 0.01) and Giardia and nitrate (r = 0.502, p < 0.01). This study was aimed to create greater awareness of parasitic contamination in the water environment in the Philippines and also to act as a platform of the current scenario for policymakers as water pollution is a key health issue in this region.

Waterborne parasites: a current status from the Philippines

PubMed Central

2014-01-01

Background Despite the amount of awareness created, waterborne disease still poses threat, especially in developing countries. Due to the scarcity of reported data on waterborne parasites, the consumption of unsafe water prolongs. Thus, the occurrences of waterborne parasites from various samples were investigated from one of the Southeast Asian country, the Philippines. Methods A total of thirty three samples, each consisting of twelve liters, were collected and processed to obtain the sediment. Ten liters of sample each was processed to detect Cryptosporidium spp. and Giardia spp. using an immunomagnetic separation method prior to enumeration via fluorescence microscope. Meanwhile, the remaining two liters were cultured to detect Acanthamoeba and Naegleria through microscopy examination and polymerase chain reaction (PCR) analysis. Results Twelve samples (36.4%) from river (5), swimming pool (1), pond (3), rain tank (1), and natural lake (2) were positive for Cryptosporidium spp., 17 (45.5%) samples from river (9), pond (2), swimming pool (1), rain tank (1), and natural lake (4) were positive for Giardia spp. while, 13 (33.3%) samples from river (3), swimming pool (2), pond (2), dispenser (1), well (1), tap (2) and natural lake (2) were positive for Acanthamoeba spp. and 5 (18.2%) samples from river (1), natural lake (1), tap (1), dispenser (1) and mineral (1) were Naegleria spp. positive. Physical parameters such as temperature, conductivity, total dissolved solid (TDS), salinity, dissolved oxygen (DO), pH, and turbidity and chemical parameters such as ammonia, chlorine, fluoride, nitrate and nitrite were also measured. The highest chemical contamination was observed at pond 2. A good correlation was observed between Giardia and nitrite (r = 0.736, p < 0.01) and Giardia and nitrate (r = 0.502, p < 0.01). Conclusion This study was aimed to create greater awareness of parasitic contamination in the water environment in the Philippines and also to act as a platform of the current scenario for policymakers as water pollution is a key health issue in this region. PMID:24885105

Chemical contaminants in water and sediment near fish nesting sites in the Potomac River basin: determining potential exposures to smallmouth bass (Micropterus dolomieu)

USGS Publications Warehouse

Kolpin, Dana W.; Blazer, Vicki; Gray, James L.; Focazio, Michael J.; Young, John A.; Alvarez, David A.; Iwanowicz, Luke R.; Foreman, William T.; Furlong, Edward T.; Speiran, Gary K.; Zaugg, Steven D.; Hubbard, Laura E.; Meyer, Michael T.; Sandstrom, Mark W.; Barber, Larry B.

2013-01-01

The Potomac River basin is an area where a high prevalence of abnormalities such as testicular oocytes (TO), skin lesions, and mortality has been observed in smallmouth bass (SMB, Micropterus dolomieu). Previous research documented a variety of chemicals in regional streams, implicating chemical exposure as one plausible explanation for these biological effects. Six stream sites in the Potomac basin (and one out-of-basin reference site) were sampled to provide an assessment of chemicals in these streams. Potential early life-stage exposure to chemicals detected was assessed by collecting samples in and around SMB nesting areas. Target chemicals included those known to be associated with important agricultural and municipal wastewater sources in the Potomac basin. The prevalence and severity of TO in SMB were also measured to determine potential relations between chemistry and biological effects. A total of 39 chemicals were detected at least once in the discrete-water samples, with atrazine, caffeine, deethylatrazine, simazine, and iso-chlorotetracycline being most frequently detected. Of the most frequently detected chemicals, only caffeine was detected in water from the reference site. No biogenic hormones/sterols were detected in the discrete-water samples. In contrast, 100 chemicals (including six biogenic hormones/sterols) were found in a least one passive-water sample, with 25 being detected at all such samples. In addition, 46 chemicals (including seven biogenic hormones/sterols) were found in the bed-sediment samples, with caffeine, cholesterol, indole, para-cresol, and sitosterol detected in all such samples. The number of herbicides detected in discrete-water samples per site had a significant positive relation to TOrank (a nonparametric indicator of TO), with significant positive relations between TOrank and atrazine concentrations in discrete-water samples and to total hormone/sterol concentration in bed-sediment samples. Such significant correlations do not necessarily imply causation, as these chemical compositions and concentrations likely do not adequately reflect total SMB exposure history, particularly during critical life stages.

Total coliform and E. coli in public water systems using undisinfected ground water in the United States.

PubMed

Messner, Michael J; Berger, Philip; Javier, Julie

2017-06-01

Public water systems (PWSs) in the United States generate total coliform (TC) and Escherichia coli (EC) monitoring data, as required by the Total Coliform Rule (TCR). We analyzed data generated in 2011 by approximately 38,000 small (serving fewer than 4101 individuals) undisinfected public water systems (PWSs). We used statistical modeling to characterize a distribution of TC detection probabilities for each of nine groupings of PWSs based on system type (community, non-transient non-community, and transient non-community) and population served (less than 101, 101-1000 and 1001-4100 people). We found that among PWS types sampled in 2011, on average, undisinfected transient PWSs test positive for TC 4.3% of the time as compared with 3% for undisinfected non-transient PWSs and 2.5% for undisinfected community PWSs. Within each type of PWS, the smaller systems have higher median TC detection than the larger systems. All TC-positive samples were assayed for EC. Among TC-positive samples from small undisinfected PWSs, EC is detected in about 5% of samples, regardless of PWS type or size. We evaluated the upper tail of the TC detection probability distributions and found that significant percentages of some system types have high TC detection probabilities. For example, assuming the systems providing data are nationally-representative, then 5.0% of the ∼50,000 small undisinfected transient PWSs in the U.S. have TC detection probabilities of 20% or more. Communities with such high TC detection probabilities may have elevated risk of acute gastrointestinal (AGI) illness - perhaps as great or greater than the attributable risk to drinking water (6-22%) calculated for 14 Wisconsin community PWSs with much lower TC detection probabilities (about 2.3%, Borchardt et al., 2012). Published by Elsevier GmbH.

Head shop compound abuse amongst attendees of the Drug Treatment Centre Board.

PubMed

McNamara, S; Stokes, S; Coleman, N

2010-05-01

The use of "Head Shop" compounds has received much media attention lately. There is very little research in the current literature with regard to the extent of the usage of these substances amongst the drug using population in Ireland. We conducted a study to examine the extent of the usage of Mephedrone, Methylone and BZP amongst attendees of Methadone maintenance programs at the DTCB. Two hundred and nine samples in total were tested. The results showed significant usage of these compounds amongst this cohort of drug users, with 29 (13.9%) of samples tested being positive for Mephedrone, 7 (3.3%) positive for Methylone and 1 (0.5%) positive for BZP.

Bacteriological and molecular studies of Clostridium perfringens infections in newly born calves.

PubMed

Selim, A M; Elhaig, M M; Zakaria, I; Ali, A

2017-01-01

Clostridium perfringens is considered one of the important causes of calf diarrhea. Two hundred and twenty-seven clinical samples from newly born and dead diarrheic calves were examined bacteriologically and by PCR. Bacterial culture identified C. perfringens in 168 of 227 samples. A total of 144 of these isolates were lecithinase positive, indicating C. perfringens Type A. In addition, 154 isolates were positive by alpha toxin encoding gene-PCR assay. This study showed high agreement between the results of bacteriology and multiplex PCR. The multiplex PCR typed all isolates that were typed as C. perfringens Type A through bacteriologic methods, but ten samples that were lecithinase negative were positive in the multiplex PCR. The study showed the highest occurrence of C. perfringens Type A isolations from calves during the winter and autumn compared with other seasons.

Epidemiological and molecular characteristics of Chlamydia psittaci from 8 human cases of psittacosis and 4 related birds in Argentina.

PubMed

Cadario, María E; Frutos, María C; Arias, Maite B; Origlia, Javier A; Zelaya, Vanina; Madariaga, María J; Lara, Claudia S; Ré, Viviana; Cuffini, Cecilia G

In Argentina, the epidemiological and molecular characteristics of Chlamydia psittaci infections are still not sufficiently known. A total of 846 respiratory and 10 ocular samples from patients with suspected human psittacosis were tested for C. psittaci from January 2010 to March 2015. Four samples of birds related to these patients were also studied. Forty-eight samples were positive for C. psittaci by a nested PCR. The molecular characterization of twelve C. psittaci PCR-positive samples received in the National Reference Laboratory INEI-ANLIS "Dr. Carlos G. Malbrán", Buenos Aires, Argentina was performed. Eight positive samples from humans and four from birds were genotyped by ompA gene sequencing. C. psittaci genotype A was found in all human samples and in the related birds. This report contributes to our increasing knowledge of the epidemiological and molecular characteristics of C. psittaci to conduct effective surveillance of its zoonotic infections. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL.

PubMed

Tiyo, Rogerio; de Souza, Carla Zangari; Nishi, Letícia; Brustolin, Camila Fernanda; Ratti, Bianca Altrão; Falavigna Guilherme, Ana Lucia

2015-01-01

The aim of this work was to compare, from a parasitological ( Cryptosporidium spp. and Giardia duodenalis), bacteriological (total and thermotolerants coliforms) and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analysis, and five liters for physicochemical analysis. Water samples were filtered under vacuum with a kit containing a cellulose acetate membrane filter, 1.2 µm (Millipore(r), Barueri, SP, Brazil). The material retained on the membrane was mechanically extracted and analyzed by direct immunofluorescence (Merifluor(r)kit). From 20 rural properties investigated, 10 had artesian wells (40 samples), 10 had common wells (40 samples), and one had a mine (four samples), the latter contaminated by Cryptosporidium spp. In samples from artesian wells, 90 to 130 meters depth, 42.5% were positive for total coliforms and 5.0% were identified to have abnormal coloration. From the samples of common wells, 14 to 37 meters depth, 87.5% were contaminated with total coliforms, 82.5% were positive for thermotolerant coliforms, and 12.5% had color abnormalities. We did not detect the presence of Giardia spp. or Cryptosporidium spp. in artesian and common wells. The use of artesian or common wells is an important step in the control of the spreading of zoonoses, particularly Cryptosporidium spp. and Giardia spp., as well as artesian wells for coliform control in local production of vegetables to be marketed.

WATER FROM DIFFERENT SOURCES USED FOR THE IRRIGATION OF VEGETABLES TO BE MARKETED: RESEARCH ON Cryptosporidium spp., Giardia spp., AND COLIFORMS IN PARANA, BRAZIL

PubMed Central

TIYO, Rogerio; de SOUZA, Carla Zangari; NISHI, Letícia; BRUSTOLIN, Camila Fernanda; RATTI, Bianca Altrão; FALAVIGNA GUILHERME, Ana Lucia

2015-01-01

SUMMARY The aim of this work was to compare, from a parasitological ( Cryptosporidiumspp. and Giardia duodenalis), bacteriological (total and thermotolerants coliforms) and physicochemical perspective, water sources used for drinking and irrigation of vegetables intended to be sold for human consumption. From January 2010 to May 2011, samples of different water sources from vegetable producing properties were collected; 100 liters for parasitological analysis, 200 mL for bacteriological analysis, and five liters for physicochemical analysis. Water samples were filtered under vacuum with a kit containing a cellulose acetate membrane filter, 1.2 µm (Millipore(r), Barueri, SP, Brazil). The material retained on the membrane was mechanically extracted and analyzed by direct immunofluorescence (Merifluor(r)kit). From 20 rural properties investigated, 10 had artesian wells (40 samples), 10 had common wells (40 samples), and one had a mine (four samples), the latter contaminated by Cryptosporidiumspp. In samples from artesian wells, 90 to 130 meters depth, 42.5% were positive for total coliforms and 5.0% were identified to have abnormal coloration. From the samples of common wells, 14 to 37 meters depth, 87.5% were contaminated with total coliforms, 82.5% were positive for thermotolerant coliforms, and 12.5% had color abnormalities. We did not detect the presence of Giardiaspp. or Cryptosporidiumspp. in artesian and common wells. The use of artesian or common wells is an important step in the control of the spreading of zoonoses, particularly Cryptosporidiumspp. and Giardiaspp., as well as artesian wells for coliform control in local production of vegetables to be marketed. PMID:26422158

[Analysis on the results of etiology and serology of plague in Qinghai province from 2001 to 2010].

PubMed

Yang, Yonghai; Wang, Mei; Zhao, Xiaolong; Zhao, Zhongzhi; Zhang, Aiping; Wei, Rongjie; Wei, Baiqing; Wang, Zuyun

2014-02-01

To analyze the results of etiology and serology of plague among human and infected animals in Qinghai province from 2001 to 2010. Thirty-seven cases of human infected with plague, 53 541 different animal samples, 5 685 sets of vector insects flea and 49 039 different animal serum samples were obtained between 2001 and 2010. A total of 7 811 samples of serum from healthy farmers and herdsmen in 14 counties in Qinghai from 2005 to 2007 were collected. Yersinia pestis (Y. pestis) were detected in visceral and secretions from human, infected animals and vector insects, respectively. Plague antigen was detected by reverse indirect hemagglutination assay (RIHA) in those samples. Indirect hemagglutination assay (IHA) was used to test plague FI antibody in serum of human and infected animals. 37 human plague cases were confirmed, 21 strains of plague Y. pestis were isolated from human cases and 14 positive were detected out. 133 of 7 811 samples of human serum were IHA positive, with the positive rate at 1.7%. A total of 146 strains of plague were isolated from infected animals and vector insects, 99 out of which were from infected animals, with a ratio of Marmota himalayan at 72.7% (72/99) and the other 47 were from vector insects, with a ratio of callopsylla solaris at 68.1% (32/47). The number of IHA and PIHA positive were 300 and 10, respectively. A total of 3 animals and 3 insects species were identified as new epidemic hosts for plague. The natural plague focus of Microtus fuscus was discovered and confirmed and coexisted with natural focus of Marmota himalayan in Chengduo county, Yushu prefecture. The epidemic situation of plague is distributed mainly in Haixi, Yushu and Hainan prefectures. From 2001 to 2010, animal infected with plague was detected in successive years and human plague was very common in Qinghai. New infected animals and vector insects species and new epidemic areas were confirmed, hence the trend of plague prevalence for humans and animals is very active in Qinghai province.

Molecular characterizations of Vibrio parahaemolyticus in seafood from the Black Sea, Turkey.

PubMed

Terzi Gulel, G; Martinez-Urtaza, J

2016-06-01

Vibrio parahaemolyticus is a marine bacterium that is considered as one of the major causes of bacterial food-borne outbreaks at a global scale. A total of 114 samples including mussel (n = 42), seawater (n = 22) and fish (n = 50) samples were collected and subjected to investigation. Vibrio parahaemolyticus was detected in 45 (39%) of 114 samples with an occurrence in mussel, seawater and fish samples of 76, 40·9 and 8% respectively. A total of 96 isolates were positive for the species-specific genes toxR and tlh and confirmed as V. parahaemolyticus. Presence of the virulence marker gene tdh was not identified in any of the strains investigated; however, four of strains were positive for the trh gene. Serological analysis of eight randomly selected trh-negative isolates identified three different serotypes: O4:K untypeable (KUT), O2:KUT, O3:KUT. Conversely, all four trh-positive strains belonged to a single serotype (O1:K1) and share an undistinguishable genetic profile by PFGE analysis, suggesting the existence of a dominant clone for the trh-positive strains in the region. Vibrio parahaemolyticus is the most prevalent food-poisoning bacterium associated with seafood consumption. The number of infections is increasing worldwide and is being reported in areas with no previous incidence. This study provides the first instance of the occurrence of V. parahaemolyticus strains with virulence traits in the Black Sea, contributing to gain a better understanding about potential risk associated with this pathogen in the region. © 2016 The Society for Applied Microbiology.

COMPARISON OF THE RECOVERIES OF ESCHERICHIA COLI AND TOTAL COLIFORMS FROM DRINKING WATER BY THE MI AGAR METHOD AND THE U.S. ENVIRONMENTAL PROTECTION AGENCY-APPROVED MEMBRANE FILTER METHOD

EPA Science Inventory

Drinking water regulations under the Final Coliform Rule require that total coliform-positive drinking water samples be examined for the presence of Escherichia coli or fecal coliforms. The current U.S. Environmental Protection Agency-approved membrane filter (MF) method for E. c...

Detection and identification of Leishmania spp.: application of two hsp70-based PCR-RFLP protocols to clinical samples from the New World.

PubMed

Montalvo, Ana M; Fraga, Jorge; Tirado, Dídier; Blandón, Gustavo; Alba, Annia; Van der Auwera, Gert; Vélez, Iván Darío; Muskus, Carlos

2017-07-01

Leishmaniasis is highly prevalent in New World countries, where several methods are available for detection and identification of Leishmania spp. Two hsp70-based PCR protocols (PCR-N and PCR-F) and their corresponding restriction fragment length polymorphisms (RFLP) were applied for detection and identification of Leishmania spp. in clinical samples recruited in Colombia, Guatemala, and Honduras. A total of 93 cases were studied. The samples were classified into positive or suspected of leishmaniasis according to parasitological criteria. Molecular amplification of two different hsp70 gene fragments and further RFLP analysis for identification of Leishmania species was done. The detection in parasitologically positive samples was higher using PCR-N than PCR-F. In the total of samples studied, the main species identified were Leishmania panamensis, Leishmania braziliensis, and Leishmania infantum (chagasi). Although RFLP-N was more efficient for the identification, RFLP-F is necessary for discrimination between L. panamensis and Leishmania guyanesis, of great importance in Colombia. Unexpectedly, one sample from this country revealed an RFLP pattern corresponding to Leishmania naiffi. Both molecular variants are applicable for the study of clinical samples originated in Colombia, Honduras, and Guatemala. Choosing the better tool for each setting depends on the species circulating. More studies are needed to confirm the presence of L. naiffi in Colombian territory.

Prevalence of Listeria monocytogenes in Idiazabal cheese.

PubMed

Arrese, E; Arroyo-Izaga, M

2012-01-01

Raw-milk cheese has been identified in risk assessment as a food of greater concern to public health due to listeriosis. To determine the prevalence and levels of Listeria monocytogenes in semi-hard Idiazabal cheese manufactured by different producers in the Basque Country at consumer level. A total of 51 Idiazabal cheese samples were obtained from 10 separate retail establishments, chosen by stratified random sampling. Samples were tested using the official standard ISO procedure 11290-1 for detection and enumeration methods. All cheese samples tested negative for L. monocytogenes. However, 9.8% tested positive for Listeria spp., different from L. monocytogenes. Positive samples came from two brands, two were natural and three were smoked. The presence of Listeria spss. suggests that the cheese making process and the hygiene whether at milking or during cheese making could be insufficient.

Prevalence of Clostridium difficile in uncooked ground meat products from Pittsburgh, Pennsylvania.

PubMed

Curry, Scott R; Marsh, Jane W; Schlackman, Jessica L; Harrison, Lee H

2012-06-01

The prevalence of Clostridium difficile in retail meat samples has varied widely. The food supply may be a source for C. difficile infections. A total of 102 ground meat and sausage samples from 3 grocers in Pittsburgh, PA, were cultured for C. difficile. Brand A pork sausages were resampled between May 2011 and January 2012. Two out of 102 (2.0%) meat products initially sampled were positive for C. difficile; both were pork sausage from brand A from the same processing facility (facility A). On subsequent sampling of brand A products, 10/19 samples from processing facility A and 1/10 samples from 3 other facilities were positive for C. difficile. The isolates recovered were inferred ribotype 078, comprising 6 genotypes. The prevalence of C. difficile in retail meat may not be as high as previously reported in North America. When contamination occurs, it may be related to events at processing facilities.

Prevalence and distribution of Arcobacter spp. in raw milk and retail raw beef.

PubMed

Shah, A H; Saleha, A A; Murugaiyah, M; Zunita, Z; Memon, A A

2012-08-01

A total of 106 beef samples which consisted of local (n = 59) and imported (n = 47) beef and 180 milk samples from cows (n = 86) and goats (n = 94) were collected from Selangor, Malaysia. Overall, 30.2% (32 of 106) of beef samples were found positive for Arcobacter species. Imported beef was significantly more contaminated (46.80%) than local beef (16.9%). Arcobacter butzleri was the species isolated most frequently from imported (81.8%) and local (60%) beef, followed by Arcobacter cryaerophilus in local (33.3%) and imported (18.2%) beef samples. Only one local beef sample (10%) yielded Arcobacter skirrowii. Arcobacter species were detected from cow's milk (5.8%), with A. butzleri as the dominant species (60%), followed by A. cryaerophilus (40%), whereas none of the goat's milk samples were found positive for Arcobacter. This is the first report of the detection of Arcobacter in milk and beef in Malaysia.

The association between total phthalate concentration and non-communicable diseases and chronic inflammation in South Australian urban dwelling men.

PubMed

Bai, Peter Y; Wittert, Gary; Taylor, Anne W; Martin, Sean A; Milne, Robert W; Jenkins, Alicia J; Januszewski, Andrzej S; Shi, Zumin

2017-10-01

To investigate associations between urinary total phthalate concentration, chronic low-grade inflammation and non-communicable diseases in a cohort of South Australian men. 1504 men aged 39-84 years who provided a urinary sample at the follow-up visit of the Men Androgen Inflammation Lifestyle Environment and Stress (MAILES) study, a randomly-selected group of urban-dwelling, community-based men from Adelaide, Australia (n = 2038; study participation rate: 78.1%). Total phthalate concentration was quantified in fasting morning urine samples. Chronic diseases were assessed through self-report questionnaire or directly measured using standardised clinical and laboratory procedures. Inflammatory biomarkers were assayed by ELISA or spectroscopy. Multivariable linear and logistic regression models were applied to determine associations of log-transformed urinary phthalate concentration with inflammation and chronic disease. Total phthalates were detected in 99.6% of urinary samples; geometric mean (95% CI) was 114.1 (109.5-118.9)µg/g creatinine. Higher total phthalate levels were associated with higher levels of hs-CRP, IL-6 (all p < 0.05) and TNF-α but not MPO. Urinary total phthalate concentrations were positively associated with cardiovascular disease, type-2-diabetes and hypertension. Comparing extreme quartiles of total phthalate, prevalence ratios were 1.78 (95% CI 1.17 - 2.71, p-trend = 0.001) for cardiovascular disease and 1.84 (95%CI 1.34 - 2.51, p-trend = 0.001) for type-2-diabetes and 1.14 (95%CI 1.01 - 1.29, p-trend = 0.013) for hypertension. Total phthalates and asthma and depression were not significantly associated. A positive association between total phthalates and cardiovascular disease, type-2-diabetes, hypertension and increased levels of chronic low-grade inflammatory biomarkers was observed in urban-dwelling Australian men. Copyright © 2017 Elsevier Inc. All rights reserved.

Adenovirus is a key pathogen in hemorrhagic cystitis associated with bone marrow transplantation.

PubMed

Akiyama, H; Kurosu, T; Sakashita, C; Inoue, T; Mori Si; Ohashi, K; Tanikawa, S; Sakamaki, H; Onozawa, Y; Chen, Q; Zheng, H; Kitamura, T

2001-05-01

Late-onset hemorrhagic cystitis (HC) is a well-known complication of bone marrow transplantation (BMT) that is mainly attributed to infection with BK virus (BKV) and adenovirus (AdV). From 1986 through 1998, 282 patients underwent BMT, and 45 of them developed HC. Urine samples tested positive for AdV in 26 patients, of which 22 showed virus type 11. Among patients who underwent allogeneic BMT, logistic regression analysis revealed acute graft-versus-host disease (grade, > or = 2) to be the most significant predictive factor for HC (P < .0001). In addition, a total of 193 urine samples regularly obtained from 26 consecutive patients who underwent allogeneic BMT were examined for BKV, JC virus (JCV), and AdV by means of polymerase chain reaction. Of patients without HC, approximately 30% of the specimens tested positive for BKV (58 samples) and JCV (55 samples), whereas 5 (3%) tested positive for AdV. Of the 3 samples obtained from patients with HC, the numbers of positive results for BKV, JCV, and AdV were 3, 1, and 1, respectively; the numbers of positive results increased to 14 of 17, 9 of 17, and 10 of 17, respectively, when we added another 14 samples obtained from 14 patients with HC (P < .0001, P = .026, and P < .0001, respectively). In conclusion, there was significant correlation between AdV and HC in the patients we studied.

Source apportionment of heavy metals and ionic contaminants in rainwater tanks in a subtropical urban area in Australia.

PubMed

Huston, R; Chan, Y C; Chapman, H; Gardner, T; Shaw, G

2012-03-15

Due to prolonged droughts in recent years, the use of rainwater tanks in urban areas has increased in Australia. In order to apportion sources of contribution to heavy metal and ionic contaminants in rainwater tanks in Brisbane, a subtropical urban area in Australia, monthly tank water samples (24 sites, 31 tanks) and concurrent bulk deposition samples (18 sites) were collected during mainly April 2007-March 2008. The samples were analysed for acid-soluble metals, soluble anions, total inorganic carbon and total organic carbon, and characteristics such as total solid and pH. The Positive Matrix Factorisation model, EPA PMF 3.0, was used to apportion sources of contribution to the contaminants. Four source factors were identified for the bulk deposition samples, including 'crustal matter/sea salt', 'car exhausts/road side dust', 'industrial dust' and 'aged sea salt/secondary aerosols'. For the tank water samples, apart from these atmospheric deposition related factors which contributed in total to 65% of the total contaminant concentration on average, another six rainwater collection system related factors were identified, including 'plumbing', 'building material', 'galvanizing', 'roofing', 'steel' and 'lead flashing/paint' (contributing in total to 35% of the total concentration on average). The Australian Drinking Water Guideline for lead was exceeded in 15% of the tank water samples. The collection system related factors, in particular the 'lead flashing/paint' factor, contributed to 79% of the lead in the tank water samples on average. The concentration of lead in tank water was found to vary with various environmental and collection system factors, in particular the presence of lead flashing on the roof. The results also indicated the important role of sludge dynamics inside the tank on the quality of tank water. Copyright © 2011 Elsevier Ltd. All rights reserved.

A rapid chemiluminescent slot blot immunoassay for the detection and quantification of Clostridium botulinum neurotoxin type E, in cultures.

PubMed

Cadieux, Brigitte; Blanchfield, Burke; Smith, James P; Austin, John W

2005-05-01

A simple, rapid, cost-effective in vitro slot blot immunoassay was developed for the detection and quantification of botulinum neurotoxin type E (BoNT/E) in cultures. Culture supernatants of 36 strains of clostridia, including 12 strains of Clostridium botulinum type E, 12 strains of other C. botulinum neurotoxin serotypes, and 12 strains of other clostridial species were tested. Samples containing BoNT/E were detected using affinity-purified polyclonal rabbit antisera prepared against BoNT/E with subsequent detection of secondary antibodies using chemiluminescence. All strains of C. botulinum type E tested positive, while all non C. botulinum type E strains tested negative. The sensitivity of the slot blot immunoassay for detection of BoNT/E was approximately four mouse lethal doses (MLD). The intensity of chemiluminescence was directly correlated with the concentration of BoNT/E up to 128 MLD, allowing quantification of BoNT/E between 4 and 128 MLD. The slot blot immunoassay was compared to the mouse bioassay for detection of BoNT/E using cultures derived from fish samples inoculated with C. botulinum type E, and cultures derived from naturally contaminated environmental samples. A total of 120 primary enrichment cultures derived from fish samples, of which 103 were inoculated with C. botulinum type E, and 17 were uninoculated controls, were assayed. Of the 103 primary enrichment cultures derived from inoculated fish samples, all were positive by mouse bioassay, while 94 were also positive by slot blot immunoassay, resulting in a 7.5% false-negative rate. All 17 primary enrichment cultures derived from the uninoculated fish samples were negative by both mouse bioassay and slot blot immunoassay. A total of twenty-six primary enrichment cultures derived from environmental samples were tested by mouse bioassay and slot blot immunoassay. Of 13 primary enrichment cultures positive by mouse bioassay, 12 were also positive by slot blot immunoassay, resulting in a 3.8% false-negative rate. All 13 primary enrichment cultures that tested negative by mouse bioassay also tested negative by slot blot immunoassay. The slot blot immunoassay could be used routinely as a positive screen for BoNT/E in primary enrichment cultures, and could be used as a replacement for the mouse bioassay for pure cultures.

Viable Mycobacterium avium ssp. paratuberculosis isolated from calf milk replacer.

PubMed

Grant, Irene R; Foddai, Antonio C G; Tarrant, James C; Kunkel, Brenna; Hartmann, Faye A; McGuirk, Sheila; Hansen, Chungyi; Talaat, Adel M; Collins, Michael T

2017-12-01

When advising farmers on how to control Johne's disease in an infected herd, one of the main recommendations is to avoid feeding waste milk to calves and instead feed calf milk replacer (CMR). This advice is based on the assumption that CMR is free of viable Mycobacterium avium ssp. paratuberculosis (MAP) cells, an assumption that has not previously been challenged. We tested commercial CMR products (n = 83) obtained from dairy farms around the United States by the peptide-mediated magnetic separation (PMS)-phage assay, PMS followed by liquid culture (PMS-culture), and direct IS900 quantitative PCR (qPCR). Conventional microbiological analyses for total mesophilic bacterial counts, coliforms, Salmonella, coagulase-negative staphylococci, streptococci, nonhemolytic Corynebacterium spp., and Bacillus spp. were also performed to assess the overall microbiological quality of the CMR. Twenty-six (31.3%) of the 83 CMR samples showed evidence of the presence of MAP. Seventeen (20.5%) tested positive for viable MAP by the PMS-phage assay, with plaque counts ranging from 6 to 1,212 pfu/50 mL of reconstituted CMR (average 248.5 pfu/50 mL). Twelve (14.5%) CMR samples tested positive for viable MAP by PMS-culture; isolates from all 12 of these samples were subsequently confirmed by whole-genome sequencing to be different cattle strains of MAP. Seven (8.4%) CMR samples tested positive for MAP DNA by IS900 qPCR. Four CMR samples tested positive by both PMS-based tests and 5 CMR samples tested positive by IS900 qPCR plus one or other of the PMS-based tests, but only one CMR sample tested positive by all 3 MAP detection tests applied. All conventional microbiology results were within current standards for whole milk powders. A significant association existed between higher total bacterial counts and presence of viable MAP indicated by either of the PMS-based assays. This represents the first published report of the isolation of viable MAP from CMR. Our findings raise concerns about the potential ability of MAP to survive manufacture of dried milk-based products. The Authors. Published by the Federation of Animal Science Societies and Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

[Diagnostic yield of paediatric respiratory samples in the Balearic Islands Sentinel Influenza Surveillance Network].

PubMed

Reina, J; Nicolau, A; Galmes, A; Arbona, B

2009-05-01

Influenza disease is subjected to surveillance by national networks (RC) that predict the epidemic behaviour by reporting clinical and virological data. To evaluate the effectiveness of the paediatric respiratory samples in the Balearic Islands RC in the last five epidemic seasons. A breath sample was taken from paediatric patients in the RC who had flu symptoms. The samples were inoculated in the MDCK cell line. We reviewed the epidemiological data of patients with a culture positive to influenza A and B. A total of 338 pharyngeal swabs from the RC were analysed during the study period. Of these, 65 (19.3%) belonged to <14 years old patients, and 44.6% of the samples were positive as opposed to 39.1% of adult respiratory samples. The influenza A virus was isolated in 24 paediatric samples (82.7%) and the influenza B virus in 5 (17.3%). The mean age of the paediatric patients of the RC who were positive was 8.5 years. Only 3 patients in the 0-4 year old group were positive (10.3%) and 26 patients (89.7%) in the 5-14 years old group. In spite that paediatricians represented only 22% of the RC doctors and obtain the 19.3% of all respiratory samples, the percentage and effectiveness of these is higher that that obtained in the adult population.

Responses of aquatic macrophytes to anthropogenic pressures: comparison between macrophyte metrics and indices.

PubMed

Camargo, Julio A

2018-02-26

Macrophyte responses to anthropogenic pressures in two rivers of Central Spain were assessed to check if simple metrics can exhibit a greater discriminatory and explanatory power than complex indices at small spatial scales. Field surveys were undertaken during the summer of 2014 (Duraton River) and the spring of 2015 (Tajuña River). Aquatic macrophytes were sampled using a sampling square (45 × 45 cm). In the middle Duraton River, macrophytes responded positively to the presence of a hydropower dam and a small weir, with Myriophyllum spicatum and Potamogeton pectinatus being relatively favored. Index of Macrophytes (IM) was better than Macroscopic Aquatic Vegetation Index (MAVI) and Fluvial Macrophyte Index (FMI) in detecting these responses, showing positive and significant correlations with total coverage, species richness, and species diversity. In the upper Tajuña River, macrophytes responded both negatively and positively to the occurrence of a trout farm effluent and a small weir, with Leptodictyum riparium and Veronica anagallis-aquatica being relatively favored. Although IM, MAVI, and FMI detected both negative and positive responses, correlations of IM with total coverage, species richness, and species diversity were higher. Species evenness was not sensitive enough to detect either positive or negative responses of aquatic macrophytes along the study areas. Overall, traditional and simple metrics (species composition, total coverage, species richness, species diversity) exhibited a greater discriminatory and explanatory power than more recent and complex indices (IM, MAVI, FMI) when assessing responses of aquatic macrophytes to anthropogenic pressures at impacted specific sites.

Can routine automated urinalysis reduce culture requests?

PubMed

Kayalp, Damla; Dogan, Kubra; Ceylan, Gozde; Senes, Mehmet; Yucel, Dogan

2013-09-01

There are a substantial number of unnecessary urine culture requests. We aimed to investigate whether urine dipstick and microscopy results could accurately rule out urinary tract infection (UTI) without urine culture. The study included a total of 32,998 patients (11,928 men and 21,070 women, mean age: 39 ± 32 years) with a preliminary diagnosis of UTI and both urinalysis and urinary culture were requested. All urine cultures were retrospectively reviewed; association of culture positivity with a positive urinalysis result for leukocyte esterase (LE) and nitrite in chemical analysis and pyuria (WBC) and bacteriuria in microscopy was determined. Diagnostic performance of urinalysis parameters for detection of UTI was evaluated. In total, 758 (2.3%) patients were positive by urine culture. Out of these culture positive samples, ratios of positive dipstick results for LE and nitrite were 71.0% (n=538) and 17.7% (n=134), respectively. The positive microscopy results for WBC and bacteria were 68.2% (n=517) and 78.8% (n=597), respectively. Negative predictive values for LE, nitrite, bacteriuria and WBC were very close to 100%. Most of the samples have no or insignificant bacterial growth. Urine dipstick and microscopy can accurately rule out UTI. Automated urinalysis is a practicable and faster screening test which may prevent unnecessary culture requests for majority of patients. © 2013. Published by Elsevier Inc. All rights reserved.

Hexavalent and total chromium at low reporting concentrations in source-water aquifers and surface waters used for public supply in Illinois, 2013

USGS Publications Warehouse

Mills, Patrick C.; Cobb, Richard P.

2015-01-01

There was a weak positive relation (ρ = 0.23) between concentrations of Cr(VI) and Cr(T) in untreated water samples, with a much stronger positive relation (ρ = 0.86 and ρ = 0.90, respectively) in samples collected soon after treatment and near the endpoint of distribution. The stronger relation and greater similarity between Cr(VI) and Cr(T) concentrations in treated water samples indicate that Cr(VI) represents a greater proportion of the measured concentrations of Cr(T) in treated waters than in untreated waters. The analysis of spikes and other quality-assurance samples indicate uncertainties associated with obtaining or confirming consistently accurate analytical results for Cr(VI) at near the applied reporting limit of 0.02 µg/L.

Low Zika virus seroprevalence among pregnant women in North Central Nigeria, 2016.

PubMed

Mathé, Philipp; Egah, Daniel Z; Müller, Janis A; Shehu, Nathan Y; Obishakin, Emmanuel T; Shwe, David D; Pam, Victor C; Okolo, Mark O; Yilgwan, Christopher; Gomerep, Simji S; Fuchs, Jonas; Abok, Ibrahim; Onyedibe, Kenneth I; Olugbo, Ewa J; Isa, Samson E; Machunga-Mambula, Salamatu S; Attah, Caleb J; Münch, Jan; Oguche, Stephen; Panning, Marcus

2018-05-26

Zika virus (ZIKV) has been known for decades in Africa but contemporary data is lacking at large. To describe the seroepidemiology of ZIKV in North Central Nigeria. We performed a cross-sectional study at six health care facilities in North Central Nigeria from January to December 2016. Detection of ZIKV antibodies was done using an anti-ZIKV recombinant non-structural protein 1 (NS1)-based ELISA. A colorimetric assay to detect ZIKV neutralizing antibodies was used on ELISA reactive and randomly selected ELISA non-reactive samples. ZIKV real-time RT-PCR was done on a subset of samples. A total of 468 individual samples were included with almost 60% from pregnant women. Using NS1-based ELISA, an anti-ZIKV positive rate of 6% for IgM and 4% for IgG was found. Pregnant women showed anti-ZIKV positive rates of 4% for IgM and 3% for IgG. None of the ZIKV antibody positive samples tested ZIKV RT-PCR positive. An association with male sex was found for anti-ZIKV IgG ELISA positivity (prevalence ratio 3.49; 95% confidence interval: 1.48-8.25; p = .004). No association with pregnancy, yellow fever vaccination or malaria was found for anti-ZIKV IgM or IgG positivity. ZIKV neutralizing antibodies were detected in 17/18 (94%) anti-ZIKV NS1 positive/borderline samples and in one sample without detectable ZIKV NS1 antibodies. Partial ZIKV E gene sequence was retrieved in one sample without ZIKV antibodies, which clustered within the West African ZIKV lineage. Our results show a largely ZIKV immunologically naïve population and reinforce the importance of ZIKV surveillance in Africa. Copyright © 2018 Elsevier B.V. All rights reserved.

The Kjeldahl method as a primary reference procedure for total protein in certified reference materials used in clinical chemistry. I. A review of Kjeldahl methods adopted by laboratory medicine.

PubMed

Chromý, Vratislav; Vinklárková, Bára; Šprongl, Luděk; Bittová, Miroslava

2015-01-01

We found previously that albumin-calibrated total protein in certified reference materials causes unacceptable positive bias in analysis of human sera. The simplest way to cure this defect is the use of human-based serum/plasma standards calibrated by the Kjeldahl method. Such standards, commutative with serum samples, will compensate for bias caused by lipids and bilirubin in most human sera. To find a suitable primary reference procedure for total protein in reference materials, we reviewed Kjeldahl methods adopted by laboratory medicine. We found two methods recommended for total protein in human samples: an indirect analysis based on total Kjeldahl nitrogen corrected for its nonprotein nitrogen and a direct analysis made on isolated protein precipitates. The methods found will be assessed in a subsequent article.

Cell block samples from malignant pleural effusion might be valid alternative samples for anaplastic lymphoma kinase detection in patients with advanced non-small-cell lung cancer.

PubMed

Zhou, Jianya; Yao, Hongtian; Zhao, Jing; Zhang, Shumeng; You, Qihan; Sun, Ke; Zou, Yinying; Zhou, Caicun; Zhou, Jianying

2015-06-01

To evaluate the clinical value of cell block samples from malignant pleural effusion (MPE) as alternative samples to tumour tissue for anaplastic lymphoma kinase (ALK) detection in patients with advanced non-small-cell lung cancer (NSCLC). Fifty-two matched samples were eligible for analysis. ALK status was detected by Ventana immunohistochemistry (IHC) (with the D5F3 clone), reverse transcription polymerase chain reaction (RT-PCR) and fluorescence in-situ hybridization (FISH) in MPE cell block samples, and by FISH in tumour tissue block samples. In total, ALK FISH results were obtained for 52 tumour tissue samples and 41 MPE cell block samples. Eight cases (15.4%) were ALK-positive in tumour tissue samples by FISH, and among matched MPE cell block samples, five were ALK-positive by FISH, seven were ALK-positive by RT-PCR, and eight were ALK-positive by Ventana IHC. The ALK status concordance rates between tumour tissue and MPE cell block samples were 78.9% by FISH, 98.1% by RT-PCR, and 100% by Ventana IHC. In MPE cell block samples, the sensitivity and specificity of Ventana IHC (100% and 100%) and RT-PCR (87.5% and 100%) were higher than those of FISH (62.5% and 100%). Malignant pleural effusion cell block samples had a diagnostic performance for ALK detection in advanced NSCLC that was comparable to that of tumour tissue samples. MPE cell block samples might be valid alternative samples for ALK detection when tissue is not available. Ventana IHC could be the most suitable method for ALK detection in MPE cell block samples. © 2014 John Wiley & Sons Ltd.

Carabelli's trait in contemporary Slovenes and inhabitants of a medieval settlement (Sredisce by the Drava River).

PubMed

Stamfelj, Iztok; Stefancić, Marija; Gaspersic, Dominik; Cvetko, Erika

2006-06-01

The objectives of this study were to determine the total frequency, expression and asymmetry of Carabelli's trait in permanent dentitions of contemporary Slovenes and a medieval skeletal population from northeastern Slovenia. A total of 254 dental casts from contemporary Slovene children were examined. The population of a medieval settlement (10th-15th centuries), was represented by 94 skeletons. A modification of the method of Alvesalo and associates was used to classify Carabelli's trait on a five-grade scale. The trait was expressed on the upper first molars of 79.7% of the contemporary subjects and 75.8% of the medieval sample. Positive expressions of the trait were found in 10.1% of the contemporary subjects and 15.2% of the medieval sample. While the observed total frequency of the trait in both samples is characteristic of Europeans, the rates of positive expressions are surprisingly low but consistent with data from a recently published worldwide literature survey. Both populations showed a low rate of left-right fluctuating asymmetry of the trait. This finding might reflect a pronounced ability of individuals in the medieval population to buffer unfavourable influences from the environment and a relatively low level of environmental stress in the contemporary population.

Prolonged incubation time does not increase sensitivity for the diagnosis of implant-related infection using samples prepared by sonication of the implants.

PubMed

Esteban, J; Alvarez-Alvarez, B; Blanco, A; Fernández-Roblas, R; Gadea, I; Garcia-Cañete, J; Sandoval, E; Valdazo, M

2013-07-01

We have designed a prospective study to evaluate the usefulness of prolonged incubation of cultures from sonicated orthopaedic implants. During the study period 124 implants from 113 patients were processed (22 osteosynthetic implants, 46 hip prostheses, 54 knee prostheses, and two shoulder prostheses). Of these, 70 patients had clinical infection; 32 had received antibiotics at least seven days before removal of the implant. A total of 54 patients had sonicated samples that produced positive cultures (including four patients without infection). All of them were positive in the first seven days of incubation. No differences were found regarding previous antibiotic treatment when analysing colony counts or days of incubation in the case of a positive result. In our experience, extending incubation of the samples to 14 days does not add more positive results for sonicated orthopaedic implants (hip and knee prosthesis and osteosynthesis implants) compared with a conventional seven-day incubation period.

40 CFR 141.859 - Coliform treatment technique triggers and assessment requirements for protection against...

Code of Federal Regulations, 2014 CFR

2014-07-01

... per month, the system has two or more total coliform-positive samples in the same month. (iii) The... second Level 1 trigger as defined in paragraph (a)(1) of this section, within a rolling 12-month period...) Level 1 treatment technique triggers. (i) For systems taking 40 or more samples per month, the system...

40 CFR 141.859 - Coliform treatment technique triggers and assessment requirements for protection against...

Code of Federal Regulations, 2013 CFR

2013-07-01

... per month, the system has two or more total coliform-positive samples in the same month. (iii) The... second Level 1 trigger as defined in paragraph (a)(1) of this section, within a rolling 12-month period...) Level 1 treatment technique triggers. (i) For systems taking 40 or more samples per month, the system...

Occurrence of infectious laryngotracheitis outbreaks in commercial layer hens detected by ELISA.

PubMed

Aras, Zeki; Yavuz, Orhan; Sanioğlu Gölen, Gökçenur

2018-02-09

Infectious laryngotracheitis (ILT) is an acute respiratory disease of chickens and a cause of great economic loss in commercial layers. The aims of this study were to investigate the prevalence of ILT in the field outbreaks and to compare the characteristics of ILT-infected and free flocks of commercial layers. A total of 625 blood serum samples were collected from 25 different layer flocks. The presence of antibodies against infectious laryngotracheitis virus (ILTV) in each sample was determined by ELISA. Of the 625 serum samples, 266 (42.56%) were found to be positive for ILTV antibodies. A total of 16 (64%) flocks were detected ILT positive by ELISA method. The mortality of infected flocks was statistically higher (P < 0.05) than uninfected flocks. The egg production of positive flocks was lower than that of the free flocks, but this difference was not statistically significant. The average live weight of hens in infected flocks was lower (P > 0.05) than hens in free flocks. In conclusion, the results of this study indicated a high prevalence of ILT infection in the commercial layer flocks in Konya region, Turkey. In outbreaks, ILT significantly increased the mortality rate and decreased the average live weight in layer hens.

Activity and Phylogenetic Diversity of Bacterial Cells with High and Low Nucleic Acid Content and Electron Transport System Activity in an Upwelling Ecosystem

PubMed Central

Longnecker, K.; Sherr, B. F.; Sherr, E. B.

2005-01-01

We evaluated whether bacteria with higher cell-specific nucleic acid content (HNA) or an active electron transport system, i.e., positive for reduction of 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), were responsible for the bulk of bacterioplankton metabolic activity. We also examined whether the phylogenetic diversity of HNA and CTC-positive cells differed from the diversity of Bacteria with low nucleic acid content (LNA). Bacterial assemblages were sampled both in eutrophic shelf waters and in mesotrophic offshore waters in the Oregon coastal upwelling region. Cytometrically sorted HNA, LNA, and CTC-positive cells were assayed for their cell-specific [3H]leucine incorporation rates. Phylogenetic diversity in sorted non-radioactively labeled samples was assayed using denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA genes. Cell-specific rates of leucine incorporation of HNA and CTC-positive cells were on average only slightly greater than the cell-specific rates of LNA cells. HNA cells accounted for most bacterioplankton substrate incorporation due to high abundances, while the low abundances of CTC-positive cells resulted in only a small contribution by these cells to total bacterial activity. The proportion of the total bacterial leucine incorporation attributable to LNA cells was higher in offshore regions than in shelf waters. Sequence data obtained from DGGE bands showed broadly similar phylogenetic diversity across HNA, LNA, and CTC-positive cells, with between-sample and between-region variability in the distribution of phylotypes. Our results suggest that LNA bacteria are not substantially different from HNA bacteria in either cell-specific rates of substrate incorporation or phylogenetic composition and that they can be significant contributors to bacterial metabolism in the sea. PMID:16332746

Activity and phylogenetic diversity of bacterial cells with high and low nucleic acid content and electron transport system activity in an upwelling ecosystem.

PubMed

Longnecker, K; Sherr, B F; Sherr, E B

2005-12-01

We evaluated whether bacteria with higher cell-specific nucleic acid content (HNA) or an active electron transport system, i.e., positive for reduction of 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), were responsible for the bulk of bacterioplankton metabolic activity. We also examined whether the phylogenetic diversity of HNA and CTC-positive cells differed from the diversity of Bacteria with low nucleic acid content (LNA). Bacterial assemblages were sampled both in eutrophic shelf waters and in mesotrophic offshore waters in the Oregon coastal upwelling region. Cytometrically sorted HNA, LNA, and CTC-positive cells were assayed for their cell-specific [3H]leucine incorporation rates. Phylogenetic diversity in sorted non-radioactively labeled samples was assayed using denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA genes. Cell-specific rates of leucine incorporation of HNA and CTC-positive cells were on average only slightly greater than the cell-specific rates of LNA cells. HNA cells accounted for most bacterioplankton substrate incorporation due to high abundances, while the low abundances of CTC-positive cells resulted in only a small contribution by these cells to total bacterial activity. The proportion of the total bacterial leucine incorporation attributable to LNA cells was higher in offshore regions than in shelf waters. Sequence data obtained from DGGE bands showed broadly similar phylogenetic diversity across HNA, LNA, and CTC-positive cells, with between-sample and between-region variability in the distribution of phylotypes. Our results suggest that LNA bacteria are not substantially different from HNA bacteria in either cell-specific rates of substrate incorporation or phylogenetic composition and that they can be significant contributors to bacterial metabolism in the sea.

Health assessment of free-living eastern box turtles (Terrapene carolina carolina) in and around the Maryland Zoo in Baltimore 1996-2011.

PubMed

Adamovicz, Laura; Bronson, Ellen; Barrett, Kevin; Deem, Sharon L

2015-03-01

Health data for free-living eastern box turtles (Terrapene carolina carolina) at the Maryland Zoo in Baltimore were analyzed. One hundred and eighteen turtles were captured on or near zoo grounds over the course of 15 yr (1996-2011), with recapture of many individuals leading to 208 total evaluations. Of the 118 individuals, 61 were male, 50 were female, and 7 were of undetermined sex. Of the 208 captures, 188 were healthy, and 20 were sick or injured. Complete health evaluations were performed on 30 turtles with physical examination records, complete blood counts (CBCs), and plasma biochemistry profiles. Eight animals were sampled more than once, yielding 40 total samples for complete health evaluations of these 30 individuals. The 40 samples were divided into healthy (N=29) and sick (N=11) groups based on clinical findings on physical examination. Samples from healthy animals were further divided into male (N=17) and female (N= 12) groups. CBC and biochemistry profile parameters were compared between sick and healthy groups and between healthy males and females. Sick turtles had lower albumin, globulin, total protein (TP), calcium, phosphorous, sodium, and potassium than healthy animals. Sick turtles also had higher heterophil to lymphocyte ratios. Healthy female turtles had higher leukocyte count, eosinophil count, total solids, TP, globulin, cholesterol, calcium, and phosphorous than healthy males. Banked plasma from all 40 samples was tested for antibodies to Mycoplasma agassizii and Mycoplasma testudineum via enzyme-linked immunosorbent assay. One sample from a clinically healthy female was antibody positive for M. agassizii; none were positive for M. testudineum. This study provides descriptive health data for eastern box turtles and CBC and biochemistry profile information for T. carolina carolina at and near the Maryland Zoo in Baltimore. It also reports low serologic evidence of exposure to mycoplasmosis.

Accelerating Sequences in the Presence of Metal by Exploiting the Spatial Distribution of Off-Resonance

PubMed Central

Smith, Matthew R.; Artz, Nathan S.; Koch, Kevin M.; Samsonov, Alexey; Reeder, Scott B.

2014-01-01

Purpose To demonstrate feasibility of exploiting the spatial distribution of off-resonance surrounding metallic implants for accelerating multispectral imaging techniques. Theory Multispectral imaging (MSI) techniques perform time-consuming independent 3D acquisitions with varying RF frequency offsets to address the extreme off-resonance from metallic implants. Each off-resonance bin provides a unique spatial sensitivity that is analogous to the sensitivity of a receiver coil, and therefore provides a unique opportunity for acceleration. Methods Fully sampled MSI was performed to demonstrate retrospective acceleration. A uniform sampling pattern across off-resonance bins was compared to several adaptive sampling strategies using a total hip replacement phantom. Monte Carlo simulations were performed to compare noise propagation of two of these strategies. With a total knee replacement phantom, positive and negative off-resonance bins were strategically sampled with respect to the B0 field to minimize aliasing. Reconstructions were performed with a parallel imaging framework to demonstrate retrospective acceleration. Results An adaptive sampling scheme dramatically improved reconstruction quality, which was supported by the noise propagation analysis. Independent acceleration of negative and positive off-resonance bins demonstrated reduced overlapping of aliased signal to improve the reconstruction. Conclusion This work presents the feasibility of acceleration in the presence of metal by exploiting the spatial sensitivities of off-resonance bins. PMID:24431210

Metabolic power and energetic costs of professional Australian Football match-play.

PubMed

Coutts, Aaron J; Kempton, Thomas; Sullivan, Courtney; Bilsborough, Johann; Cordy, Justin; Rampinini, Ermanno

2015-03-01

To compare the metabolic power demands between positional groups, and examine temporal changes in these parameters during Australian Football match-play. Longitudinal observational study. Global positioning system data were collected from 39 Australian Football players from the same club during 19 Australian Football League competition games over two seasons. A total of 342 complete match samples were obtained for analysis. Players were categorised into one of six positional groups: tall backs, mobile backs, midfielders, tall forwards, mobile forwards and rucks. Instantaneous raw velocity data obtained from the global positioning system units was exported to a customised spreadsheet which provided estimations of both speed-based (e.g. total and high-speed running distance) and derived metabolic power and energy expenditure variables (e.g. average metabolic power, high-power distance, total energy expenditure). There were significant differences between positional groups for both speed-based and metabolic power indices, with midfielders covering more total and high-speed distance, as well as greater average and overall energy expenditure compared to other positions (all p<0.001). There were reductions in total, high-speed, and high-power distance, as well as average metabolic power throughout the match (all p<0.001). Positional differences exist for both metabolic power and traditional running based variables. Generally, midfielders, followed by mobile forwards and mobile backs had greater activity profiles compared to other position groups. We observed that the reductions in most metabolic power variables during the course of the match are comparable to traditional running based metrics. This study demonstrates that metabolic power data may contribute to our understanding of the physical demands of Australian Football. Copyright © 2014 Sports Medicine Australia. Published by Elsevier Ltd. All rights reserved.

Antibiotic Screening of Urine Culture for Internal Quality Audit at Amrita Hospital, Kochi.

PubMed

Suresh, Aswathy; Gopinathan, Anusha; Dinesh, Kavitha R; Kumar, Anil

2017-07-01

Urine antimicrobial activity is a seldom analysed laboratory test which greatly impacts the quantification of urine specimens. Presence of antimicrobial activity in the urine reduces the bacterial load in these specimens. Hence, the chances of erroneously reporting insignificant bacteriuria can be reduced on analysis of the antimicrobial activity in urine. The aim of the study was to measure the antimicrobial activity of urine samples obtained from patients in a tertiary care hospital. A total of 100 urine specimens were collected from the study group. Tests like wet mount, Gram staining and culture were performed. Antimicrobial susceptibility testing was done on the bacteria isolated from each specimen. The urine specimens were reported as significant bacteriuria (>105 Colony Forming Unit (CFU)/ml) and insignificant bacteriuria (<105 CFU/ml - clean catch midstream urine; <102 CFU/ml - catheterized urine sample) according to the CFU/ml. Staphylococcus aureus ATCC ® 25923 ™ and Escherichia coli ATCC ® 25922 ™ were used to identify the presence of antimicrobial activity in the urine sample by Urine Anti-Bacterial substance Assay (UABA). McNemar test was used for statistical analysis using Statistical Package for the Social Sciences (SPSS) version 21.0. On analysis of the antimicrobial activity of urine sample with the prior antibiotic history of the patients, 17 were true positives and 43 were true negatives. Twenty six of samples with UABA positivity were culture negative and 28 samples with UABA positivity were culture positive. Sensitivity and specificity of the test was 85% and 53.8% respectively. Accuracy of the test was 60%. The p-value of UABA was <0.001. Enterobacteriaceae was the most common bacterial family isolated from the urine specimens. A total of 85% patients responded to treatment. Presence of antimicrobial activity in urine has a great impact on the interpretation of urine culture reports. Identification of urine antimicrobial activity helps in evaluating the quantification of bacterial growth reported in urine culture. It facilitates speedy recovery of patients by early administration of antibiotics.

Vaginal Prostate Specific Antigen (PSA) Is a Useful Biomarker of Semen Exposure Among HIV-Infected Ugandan Women.

PubMed

Woolf-King, Sarah E; Muyindike, Winnie; Hobbs, Marcia M; Kusasira, Adrine; Fatch, Robin; Emenyonu, Nneka; Johnson, Mallory O; Hahn, Judith A

2017-07-01

The practical feasibility of using prostate specific antigen (PSA) as a biomarker of semen exposure was examined among HIV-infected Ugandan women. Vaginal fluids were obtained with self-collected swabs and a qualitative rapid test (ABAcard ® p30) was used to detect PSA. Trained laboratory technicians processed samples on-site and positive PSA tests were compared to self-reported unprotected vaginal sex (UVS) in the last 48 h. A total of 77 women submitted 126 samples for PSA testing at up to three study visits. Of these samples, 31 % (n = 39/126) were PSA positive, and 64 % (n = 25/39) of the positive PSA samples were accompanied by self-report of no UVS at the study visit the PSA was collected. There were no reported difficulties with specimen collection, storage, or processing. These findings provide preliminary data on high levels of misreported UVS among HIV-infected Ugandan women using practically feasible methods for PSA collection and processing.

Excluding Anti-cytomegalovirus Immunoglobulin M-Positive Cord Blood Units Has a Minimal Impact on the Korean Public Cord Blood Bank Inventory

PubMed Central

Shin, Sue; Roh, Eun Youn; Oh, Sohee; Song, Eun Young; Kim, Eui Chong; Yoon, Jong Hyun

2017-01-01

Cord blood units (CBUs) for transplantation should be free of communicable disease and must contain a specific amount of total nucleated cells and CD34+ cells. Although posttransplantation cytomegalovirus (CMV) infections are from latent infection in patients, ensuring CMV-free CBUs by performing CMV-specific IgM and nucleic acid amplification testing (NAT) is one of the mandatory procedures for the safety of CBUs. However, the exclusion policies (based on these test results) vary among nations and institutions. We tested 28,000 processed CBUs between May 2006 and June 2014. The cord blood leukocytes from CMV IgM-positive samples were then subjected to NAT. The total nucleated cell and CD34+ cell counts were measured for each CBU, and the results were compared to the CMV IgM and IgG results. The seroprevalence of CMV among pregnant women was 98.1% (18,459/18,818) for IgG and 1.7% (441/25,293) for IgM. The concentration and the total number of CD34+ cells were significantly higher in CBUs from IgM-negative mothers compared to those from IgM-positive mothers (72.4/μl vs. 57.2/μl, respectively, p < 0.0001; 1.45 × 106/unit vs. 1.15 × 106/unit, respectively, p < 0.0001). Among CBUs with positive CMV IgM in their mothers' plasma or cord blood plasma, only 0.58% of the samples (3/517) had a positive NAT. The number of excluded CBUs from inventory due to positive CMV IgM in the cord blood was 54 of 18,326 (0.3%). For inventory purposes, it is appropriate to remove CBUs with positive cord blood CMV IgM findings irrespective of the NAT status as well as positive maternal CMV IgM in South Korea. PMID:27524276

Excluding Anti-cytomegalovirus Immunoglobulin M-Positive Cord Blood Units Has a Minimal Impact on the Korean Public Cord Blood Bank Inventory.

PubMed

Shin, Sue; Roh, Eun Youn; Oh, Sohee; Song, Eun Young; Kim, Eui Chong; Yoon, Jong Hyun

2017-01-24

Cord blood units (CBUs) for transplantation should be free of communicable disease and must contain a specific amount of total nucleated cells and CD34+ cells. Although posttransplantation cytomegalovirus (CMV) infections are from latent infection in patients, ensuring CMV-free CBUs by performing CMV-specific IgM and nucleic acid amplification testing (NAT) is one of the mandatory procedures for the safety of CBUs. However, the exclusion policies (based on these test results) vary among nations and institutions. We tested 28,000 processed CBUs between May 2006 and June 2014. The cord blood leukocytes from CMV IgM-positive samples were then subjected to NAT. The total nucleated cell and CD34+ cell counts were measured for each CBU, and the results were compared to the CMV IgM and IgG results. The seroprevalence of CMV among pregnant women was 98.1% (18,459/18,818) for IgG and 1.7% (441/25,293) for IgM. The concentration and the total number of CD34+ cells were significantly higher in CBUs from IgM-negative mothers compared to those from IgM-positive mothers (72.4/μl vs. 57.2/μl, respectively, p < 0.0001; 1.45 × 106/unit vs. 1.15 × 106/unit, respectively, p < 0.0001). Among CBUs with positive CMV IgM in their mothers' plasma or cord blood plasma, only 0.58% of the samples (3/517) had a positive NAT. The number of excluded CBUs from inventory due to positive CMV IgM in the cord blood was 54 of 18,326 (0.3%). For inventory purposes, it is appropriate to remove CBUs with positive cord blood CMV IgM findings irrespective of the NAT status as well as positive maternal CMV IgM in South Korea.

Comparison of four β-glucuronidase and β-galactosidase-based commercial culture methods used to detect Escherichia coli and total coliforms in water.

PubMed

Maheux, Andrée F; Dion-Dupont, Vanessa; Bouchard, Sébastien; Bisson, Marc-Antoine; Bergeron, Michel G; Rodriguez, Manuel J

2015-06-01

The MI agar, Colilert(®), Chromocult coliform(®) agar, and DC with BCIG agar chromogenic culture-based methods used to assess microbiological quality of drinking water were compared in terms of their ubiquity, sensitivity, ease of use, growth of atypical colonies and affordability. For ubiquity, 129 total coliform (representing 76 species) and 19 Escherichia coli strains were tested. Then, 635 1-L well water samples were divided into 100 mL subsamples for testing by all four methods. Test results showed that 70.5, 52.7, 36.4, and 23.3% of the non-E. coli total coliform strains and 94.7, 94.7, 89.5, and 89.5% of the 19 E. coli strains yielded a positive signal with the four methods, respectively. They also yielded a total coliform positive signal for 66.5, 51.7, 64.9, and 55.0% and an E. coli positive signal for 16.1, 14.8, 17.3, and 13.4% of the 635 well water samples tested, respectively. Results showed that Colilert(®) is the most expensive method tested in terms of reactants, yet it is the easiest to use. Large numbers of atypical colonies were also often observed on Chromocult coliform(®) and DC with BCIG, thereby challenging the target microorganism count. Thus, the MI agar method seems to be the best option for the assessment of drinking water quality.

Comparative impacts of stormwater runoff on water quality of an urban, a suburban, and a rural stream.

PubMed

Mallin, Michael A; Johnson, Virginia L; Ensign, Scott H

2009-12-01

Water quality data at 12 sites within an urban, a suburban, and a rural stream were collected contemporaneously during four wet and eight dry periods. The urban stream yielded the highest biochemical oxygen demand (BOD), orthophosphate, total suspended sediment (TSS), and surfactant concentrations, while the most rural stream yielded the highest total organic carbon concentrations. Percent watershed development and percent impervious surface coverage were strongly correlated with BOD (biochemical oxygen demand), orthophosphate, and surfactant concentrations but negatively with total organic carbon. Excessive fecal coliform abundance most frequently occurred in the most urbanized catchments. Fecal coliform bacteria, TSS, turbidity, orthophosphate, total phosphorus, and BOD were significantly higher during rain events compared to nonrain periods. Total rainfall preceding sampling was positively correlated with turbidity, TSS, BOD, total phosphorus, and fecal coliform bacteria concentrations. Turbidity and TSS were positively correlated with phosphorus, fecal coliform bacteria, BOD, and chlorophyll a, which argues for better sedimentation controls under all landscape types.

Gram-positive rods prevailing in teeth with apical periodontitis undergoing root canal treatment.

PubMed

Chávez de Paz, L E; Molander, A; Dahlén, G

2004-09-01

To identify Gram-positive rods from root canals of teeth with apical periodontitis and to examine their associations with other species. Consecutive root canal samples (RCSs) from 139 teeth undergoing root canal treatment were analyzed prospectively for cultivable microbes. Gram-positive rods in the first RCS submitted after chemo-mechanical preparation were categorised to genus level by selective media and gas-liquid chromatography (GLC), and identified to species level by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Associations between organisms were measured by odds ratios (OR). In the first samples submitted a total of 158 Gram-positive rods, 115 Gram-positive cocci, 26 Gram-negative rods and 9 Gram-negative cocci, were identified. At genus levels Gram-positive rods were classified into: Lactobacillus spp. (38%), Olsenella spp. (18%), Propionibacterium spp. (13%), Actinomyces spp. (12%), Bifidobacterium spp. (13%) and Eubacterium spp. (6%). The most frequent species were Olsenella uli, Lactobacillus paracasei and Propionibacterium propionicum. In subsequent samples taken during treatment, Gram-positive rods were also identified, although the number of strains was considerably reduced. Positive associations were observed between members of the genus lactobacilli and Gram-positive cocci (OR>2). Olsenella uli and Lactobacillus spp. predominated over other Gram-positive rods. A possible association exists between Lactobacillus spp. and Gram-positive cocci in root canals of teeth with apical periodontitis receiving treatment.

Rhinitis, Ocular, Throat and Dermal Symptoms, Headache and Tiredness among Students in Schools from Johor Bahru, Malaysia: Associations with Fungal DNA and Mycotoxins in Classroom Dust

PubMed Central

Norbäck, Dan; Hashim, Jamal Hisham; Cai, Gui-Hong; Hashim, Zailina; Ali, Faridah; Bloom, Erica; Larsson, Lennart

2016-01-01

There are few studies on rhinitis and sick building syndrome (SBS) among students in tropical countries. We studied associations between levels of five fungal DNA sequences, two mycotoxins (sterigmatocystin and verrucarol) and cat allergen (Fel d 1) levels in schools and rhinitis and other weekly SBS symptoms in the students. Fungal DNA was measured by quantitative PCR and cat allergen by ELISA. Pupils (N = 462) from eight randomly selected schools in Johor Bahru, Malaysia participated (96%). Dust samples were collected by cotton swabs and Petri dishes exposed for one week. None of the schools had a mechanical ventilation system, but all classrooms had openable windows that were kept open during lectures and indoor CO2 levels were low (mean 492 ppm; range 380–690 ppm). Weekly nasal symptoms (rhinitis) (18.8%), ocular (11.6%), throat (11.1%), dermal symptoms, headache (20.6%) and tiredness (22.1%) were common. Total fungal DNA in swab samples was associated with rhinitis (p = 0.02), ocular symptoms (p = 0.009) and tiredness (p = 0.001). There were positive associations between Aspergillus versicolor DNA in Petri dish samples, ocular symptoms (p = 0.02) and tiredness (p = 0.001). The level of the mycotoxin verrucarol (produced by Stachybotrys chartarum) in swab samples was positively associated with tiredness (p = 0.04). Streptomyces DNA in swab samples (p = 0.03) and Petri dish samples (p = 0.03) were negatively associated with tiredness. In conclusion, total fungal contamination, measured as total fungal DNA) in the classrooms, Aspergillus versicolor and verrucarol can be risk factors for rhinitis and SBS symptoms among students in the tropical country Malaysia. PMID:26829324

Rapid Multiplex PCR Assay To Identify Respiratory Viral Pathogens: Moving Forward Diagnosing The Common Cold

PubMed Central

Gordon, Sarah M; Elegino-Steffens, Diane U; Agee, Willie; Barnhill, Jason; Hsue, Gunther

2013-01-01

Upper respiratory tract infections (URIs) can be a serious burden to the healthcare system. The majority of URIs are viral in etiology, but definitive diagnosis can prove difficult due to frequently overlapping clinical presentations of viral and bacterial infections, and the variable sensitivity, and lengthy turn-around time of viral culture. We tested new automated nested multiplex PCR technology, the FilmArray® system, in the TAMC department of clinical investigations, to determine the feasibility of replacing the standard viral culture with a rapid turn-around system. We conducted a feasibility study using a single-blinded comparison study, comparing PCR results with archived viral culture results from a convenience sample of cryopreserved archived nasopharyngeal swabs from acutely ill ED patients who presented with complaints of URI symptoms. A total of 61 archived samples were processed. Viral culture had previously identified 31 positive specimens from these samples. The automated nested multiplex PCR detected 38 positive samples. In total, PCR was 94.5% concordant with the previously positive viral culture results. However, PCR was only 63.4% concordant with the negative viral culture results, owing to PCR detection of 11 additional viral pathogens not recovered on viral culture. The average time to process a sample was 75 minutes. We determined that an automated nested multiplex PCR is a feasible alternative to viral culture in an acute clinical setting. We were able to detect at least 94.5% as many viral pathogens as viral culture is able to identify, with a faster turn-around time. PMID:24052914

Determining the prevalence of cytomegalovirus infection in a cohort of preterm infants.

PubMed

Pitlick, Mitchell M; Orr, Kristin; Momany, Allison M; McDonald, Erin L; Murray, Jeffrey C; Ryckman, Kelli K

2015-01-01

Preterm birth is a global public health problem that is a significant cause of infant morbidity and mortality. Congenital cytomegalovirus (CMV) infection has been proposed as a risk factor for preterm birth, but the rate of CMV in infants born preterm is unclear. CMV is the leading infectious cause of sensorineural hearing loss, which will affect 15% - 20% of congenitally infected infants later in their childhood. 90% of infected infants are asymptomatic at birth and are not recognized as at risk for CMV-associated deficits. To determine the prevalence of CMV infection in a large cohort of preterm infants. DNA was extracted from cord blood, peripheral blood, saliva, and buccal swab samples collected from preterm infants. A total of 1200 unique DNA samples were tested for CMV using a nested PCR protocol. The proportions of preterm infants with CMV was compared by sample collection type, race, gender, and gestational age. A total of 37 infants tested positive for CMV (3.08%). After excluding twins, siblings, and infants older than two weeks at the time of sample collection, two out of 589 infants were CMV positive (0.3%), which was lower than the proportion of CMV observed in the general population. All positive samples came from buccal swabs. Our work suggests that while CMV infection may not be greater in preterm infants than in the general population, given the neurologic consequences of CMV in preterm infants, screening of this population may still be warranted. If so, our results suggest buccal swabs, collected at pregnancy or at birth, may be an ideal method for such a program.

Reservoir characteristics of coal-shale sedimentary sequence in coal-bearing strata and their implications for the accumulation of unconventional gas

NASA Astrophysics Data System (ADS)

Wang, Yang; Zhu, Yanming; Liu, Yu; Chen, Shangbin

2018-04-01

Shale gas and coalbed methane (CBM) are both considered unconventional natural gas and are becoming increasingly important energy resources. In coal-bearing strata, coal and shale are vertically adjacent as coal and shale are continuously deposited. Research on the reservoir characteristics of coal-shale sedimentary sequences is important for CBM and coal-bearing shale gas exploration. In this study, a total of 71 samples were collected, including coal samples (total organic carbon (TOC) content >40%), carbonaceous shale samples (TOC content: 6%-10%), and shale samples (TOC content <6%). Combining techniques of field emission scanning electron microscopy (FE-SEM), x-ray diffraction, high-pressure mercury intrusion porosimetry, and methane adsorption, experiments were employed to characterize unconventional gas reservoirs in coal-bearing strata. The results indicate that in the coal-shale sedimentary sequence, the proportion of shale is the highest at 74% and that of carbonaceous shale and coal are 14% and 12%, respectively. The porosity of all measured samples demonstrates a good positive relationship with TOC content. Clay and quartz also have a great effect on the porosity of shale samples. According to the FE-SEM image technique, nanoscale pores in the organic matter of coal samples are much more developed compared with shale samples. For shales with low TOC, inorganic minerals provide more pores than organic matter. In addition, TOC content has a positive relationship with methane adsorption capacity, and the adsorption capacity of coal samples is more sensitive than the shale samples to temperature.

Estimation of total bacteria by real-time PCR in patients with periodontal disease.

PubMed

Brajović, Gavrilo; Popović, Branka; Puletić, Miljan; Kostić, Marija; Milasin, Jelena

2016-01-01

Periodontal diseases are associated with the presence of elevated levels of bacteria within the gingival crevice. The aim of this study was to evaluate a total amount of bacteria in subgingival plaque samples in patients with a periodontal disease. A quantitative evaluation of total bacteria amount using quantitative real-time polymerase chain reaction (qRT-PCR) was performed on 20 samples of patients with ulceronecrotic periodontitis and on 10 samples of healthy subjects. The estimation of total bacterial amount was based on gene copy number for 16S rRNA that was determined by comparing to Ct values/gene copy number of the standard curve. A statistically significant difference between average gene copy number of total bacteria in periodontal patients (2.55 x 10⁷) and healthy control (2.37 x 10⁶) was found (p = 0.01). Also, a trend of higher numbers of the gene copy in deeper periodontal lesions (> 7 mm) was confirmed by a positive value of coefficient of correlation (r = 0.073). The quantitative estimation of total bacteria based on gene copy number could be an important additional tool in diagnosing periodontitis.

Seroprevalence and risk factors associated with bovine brucellosis in the Potohar Plateau, Pakistan.

PubMed

Ali, Shahzad; Akhter, Shamim; Neubauer, Heinrich; Melzer, Falk; Khan, Iahtasham; Abatih, Emmanuel Nji; El-Adawy, Hosny; Irfan, Muhammad; Muhammad, Ali; Akbar, Muhammad Waqas; Umar, Sajid; Ali, Qurban; Iqbal, Muhammad Naeem; Mahmood, Abid; Ahmed, Haroon

2017-01-28

The seroprevalence and risk factors of bovine brucellosis were studied at animal and herd level using a combination of culture, serological and molecular methods. The study was conducted in 253 randomly selected cattle herds of the Potohar plateau, Pakistan from which a total of 2709 serum (1462 cattle and 1247 buffaloes) and 2330 milk (1168 cattle and 1162 buffaloes) samples were collected. Data on risk factors associated with seroprevalence of brucellosis were collected through interviews using questionnaires. Univariable and multivariable random effects logistic regression models were used for identifying important risk factors at animal and herd levels. One hundred and seventy (6.3%) samples and 47 (18.6%) herds were seropositive for brucellosis by Rose Bengal Plate test. Variations in seroprevalence were observed across the different sampling sites. At animal level, sex, species and stock replacement were found to be potential risk factors for brucellosis. At herd level, herd size (≥9 animals) and insemination method used were important risk factors. The presence of Brucella DNA was confirmed with a real-time polymerase chain reaction assay (qRT-PCR) in 52.4% out of 170 serological positive samples. In total, 156 (6.7%) milk samples were positive by milk ring test. B. abortus biovar 1 was cultured from 5 positive milk samples. This study shows that the seroprevalence of bovine brucellosis is high in some regions in Pakistan. Prevalence was associated with herd size, abortion history, insemination methods used, age, sex and stock replacement methods. The infected animal may act as source of infection for other animals and for humans. The development of control strategies for bovine brucellosis through implementation of continuous surveillance and education programs in Pakistan is warranted.

Molecular characterization of Toxocara spp. eggs isolated from public parks and playgrounds in Shiraz, Iran.

PubMed

Choobineh, M; Mikaeili, F; Sadjjadi, S M; Ebrahimi, S; Iranmanesh, S

2018-05-07

Human toxocariasis, a worldwide parasitic disease, is caused by the larval stage of intestinal nematodes of dogs and cats, namely Toxocara canis and Toxocara cati. Human infection occurs by the accidental ingestion of embryonated eggs present in the soil, vegetables or on other contaminated surfaces, as well as via consumption of uncooked paratenic hosts, such as bird meat and giblets. The objective of this study was to evaluate the contamination of soil in public parks and playgrounds in Shiraz using microscopy and molecular methods. A total of 150 soil samples were collected from public parks and playgrounds in various areas of Shiraz, southern Iran. The samples were treated with saturated zinc sulphate solution, and Toxocara spp. eggs were detected by microscopic observation followed by nested polymerase chain reaction (PCR). To differentiate T. canis and T. cati eggs from each other, PCR restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS)-rDNA region by SalI endonuclease enzyme was used. PCR-sequencing was performed to confirm the results of the PCR-RFLP method. Based on the flotation results of the 150 soil samples, six (4%) were found to be positive for Toxocara spp. eggs, whereas nested-PCR showed 24 samples to be positive (16%). Based on the PCR-RFLP method and the sequence of the ITS-rDNA region, a total of 23 out of 24 isolates were confirmed as T. cati and one out of 24 as T. canis. The results showed a higher number of soil samples to be positive for Toxocara by the molecular method than microscopy, and higher T. cati infection in soil samples, which could have an important role in human infection with toxocariasis in this region.

Human papillomavirus detection and typing using a nested-PCR-RFLP assay.

PubMed

Coser, Janaina; Boeira, Thaís da Rocha; Fonseca, André Salvador Kazantzi; Ikuta, Nilo; Lunge, Vagner Ricardo

2011-01-01

It is clinically important to detect and type human papillomavirus (HPV) in a sensitive and specific manner. Development of a nested-polymerase chain reaction-restriction fragment length polymorphism (nested-PCR-RFLP) assay to detect and type HPV based on the analysis of L1 gene. Analysis of published DNA sequence of mucosal HPV types to select sequences of new primers. Design of an original nested-PCR assay using the new primers pair selected and classical MY09/11 primers. HPV detection and typing in cervical samples using the nested-PCR-RFLP assay. The nested-PCR-RFLP assay detected and typed HPV in cervical samples. Of the total of 128 clinical samples submitted to simple PCR and nested-PCR for detection of HPV, 37 (28.9%) were positive for the virus by both methods and 25 samples were positive only by nested-PCR (67.5% increase in detection rate compared with single PCR). All HPV positive samples were effectively typed by RFLP assay. The method of nested-PCR proved to be an effective diagnostic tool for HPV detection and typing.

The French Version of the Modified-Checklist for Autism in Toddlers (M-CHAT): A Validation Study on a French Sample of 24 Month-Old Children

ERIC Educational Resources Information Center

Baduel, Sophie; Guillon, Quentin; Afzali, Mohammad H.; Foudon, Nadège; Kruck, Jeanne; Rogé, Bernadette

2017-01-01

Early ASD screening has the potential to reduce delays between initial parental concerns and diagnosis, and promote early intervention. The aim of this study was to validate the M-CHAT on a French population sample of 24 month-old children. This study included a low-risk sample of 1,227 children. A total of 20 children screened positive on the…

Fourier transform infrared attenuated total reflection analysis of human hair: comparison of hair from breast cancer patients with hair from healthy subjects.

PubMed

Lyman, Donald J; Murray-Wijelath, Jacqueline

2005-01-01

A comparative study of Fourier transform infrared attenuated total reflection (FTIR-ATR) spectra of 32 scalp and pubic hair samples from individuals diagnosed with breast cancer and those who were negative for breast cancer showed increases in the beta-sheet/disorder structures (relative to alpha-helix structures) and C-H lipid content of hair from breast cancer patients. Thus, the presence of breast cancer appears to alter the hair growth process, resulting in changes in the composition and conformation of cell membrane and matrix materials of hair fiber. These appear to be consistent with the changes observed in X-ray diffraction patterns for hair from breast cancer patients. A blind study of 12 additional hair samples using these FTIR-ATR spectral differences as markers correctly identified all four hair samples from cancer patients (100%). Two of these samples were from breast cancer patients. Of the remaining two samples analyzing positive for cancer, one was from a prostate cancer patient and one from a lung cancer patient. Thus, it appears that the mechanism that alters hair fiber synthesis in the three types of cancer may be similar. The blind study incorrectly identified as positive for cancer three hair samples from two apparently healthy individuals and one patient considered cured from prostate cancer.

Five-year microbiological monitoring of wards and operating theatres in southern Italy.

PubMed

La Fauci, V; Genovese, C; Facciolà, A; Palamara, M A R; Squeri, R

2017-06-01

Nosocomial infections are one of the greatest problems in public health. Several studies have highlighted the role played by the hospital environment as a possible source of transmission of nosocomial pathogens. A five-year monitoring of bacterial contamination on healthcare workers hands, surfaces most closely in contact with inpatient wards, operating theatres and "at rest" and "in use" operating theatre air samples. For the samples, we used sterile swabs, contact slides, manual API, and automated VITEK systems for identification. In the five-year period, a total of 9396 samples were collected and analysed. In ward patients, 4398 samplings were carried out with 4.7%, 9.4%, 7%, 10.8% and 7.9% positive results respectively from 2010 to 2014. For hands, 648 samplings were carried out, with a positivity of 40.74%. In operating theatres, 4188 samples were taken, with a positivity of 11.9%. Regarding air in empty and full theatres, 1962 samplings were carried out with a positivity rate equal to 31.9%. The monitoring showed a low rate of contamination with a progressive decrease in the fiveyear period on operating theatres surfaces and hands, while there was an increase in the surgical site wards and in the air of operating rooms. Our investigation has revealed the presence of pathogens on the assessed surfaces and the need for environmental monitoring, which can be a valuable tool for reducing contamination.

PubMed Central

Genovese, C.; Facciolà, A.; Palamara, M.A.R.; Squeri, R.

2017-01-01

Summary Introduction. Nosocomial infections are one of the greatest problems in public health. Several studies have highlighted the role played by the hospital environment as a possible source of transmission of nosocomial pathogens. Methods. A five-year monitoring of bacterial contamination on healthcare workers hands, surfaces most closely in contact with inpatient wards, operating theatres and "at rest" and "in use" operating theatre air samples. For the samples, we used sterile swabs, contact slides, manual API, and automated VITEK systems for identification. Results. In the five-year period, a total of 9396 samples were collected and analysed. In ward patients, 4398 samplings were carried out with 4.7%, 9.4%, 7%, 10.8% and 7.9% positive results respectively from 2010 to 2014. For hands, 648 samplings were carried out, with a positivity of 40.74%. In operating theatres, 4188 samples were taken, with a positivity of 11.9%. Regarding air in empty and full theatres, 1962 samplings were carried out with a positivity rate equal to 31.9%. The monitoring showed a low rate of contamination with a progressive decrease in the fiveyear period on operating theatres surfaces and hands, while there was an increase in the surgical site wards and in the air of operating rooms. Conclusions. Our investigation has revealed the presence of pathogens on the assessed surfaces and the need for environmental monitoring, which can be a valuable tool for reducing contamination. PMID:28900357

Occurrence of severe gastroenteritis in pups after canine parvovirus vaccine administration: a clinical and laboratory diagnostic dilemma.

PubMed

Decaro, Nicola; Desario, Costantina; Elia, Gabriella; Campolo, Marco; Lorusso, Alessio; Mari, Viviana; Martella, Vito; Buonavoglia, Canio

2007-01-26

A total of 29 faecal samples collected from dogs with diarrhoea following canine parvovirus (CPV) vaccination were tested by minor groove binder (MGB) probe assays for discrimination between CPV vaccine and field strains and by diagnostic tests for detection of other canine pathogens. Fifteen samples tested positive only for CPV field strains; however, both vaccine and field strains were detected in three samples. Eleven samples were found to contain only the vaccine strain, although eight of them tested positive for other pathogens of dogs. Only three samples were found to contain the vaccine strain without evidence of canine pathogens. The present study confirms that most cases of parvovirus-like disease occurring shortly after vaccination are related to infection with field strains of canine parvovirus type 2 (CPV-2) rather than to reversion to virulence of the modified live virus contained in the vaccine.

Molecular Detection of Avian Influenza Virus from Sediment Samples in Waterfowl Habitats on the Delmarva Peninsula, United States.

PubMed

Densmore, C L; Iwanowicz, D D; Ottinger, C A; Hindman, L J; Bessler, A M; Iwanowicz, L R; Prosser, D J; Whitbeck, M; Driscoll, C P

2017-12-01

Avian influenza viruses (AIV) affect many species of birds including waterfowl and may persist in sediment in aquatic habitats. Sediment samples were collected from two areas representative of prime migration and overwintering waterfowl habitat in Dorchester County, Maryland in the fall and winter of 2013-2014. Samples were screened for the presence of AIV via reverse transcriptase-quantitative PCR targeting the matrix gene. Although 13.6% of sediment samples were positive for the AIV matrix gene across all collection dates and locations, differences in detection were noted with location and collection season. Percentage of AIV-positive sediment samples recovered corresponded to trends in waterfowl abundance at collection sites both temporally and spatially. These findings provide further support for the assertion that the presence of AIV in the aquatic environment is likely affected by the total number, site-specific density, and array of waterfowl species.

Variability of Total and Pathogenic Vibrio parahaemolyticus Densities in Northern Gulf of Mexico Water and Oysters▿

PubMed Central

Zimmerman, A. M.; DePaola, A.; Bowers, J. C.; Krantz, J. A.; Nordstrom, J. L.; Johnson, C. N.; Grimes, D. J.

2007-01-01

Vibrio parahaemolyticus is indigenous to coastal environments and a frequent cause of seafood-borne gastroenteritis in the United States, primarily due to raw-oyster consumption. Previous seasonal-cycle studies of V. parahaemolyticus have identified water temperature as the strongest environmental predictor. Salinity has also been identified, although it is evident that its effect on annual variation is not as pronounced. The effects of other environmental factors, both with respect to the seasonal cycle and intraseasonal variation, are uncertain. This study investigated intraseasonal variations of densities of total and pathogenic V. parahaemolyticus organisms in oysters and overlying waters during the summer of 2004 at two sites in the northern Gulf of Mexico. Regression analyses indicated significant associations (P < 0.001) between total V. parahaemolyticus densities and salinity, as well as turbidity in water and in oysters at the Mississippi site but not at the Alabama site. Pathogenic V. parahaemolyticus organisms in Mississippi oyster and water samples were detected in 56% (9 out of 16) and 78% (43 out of 55) of samples, respectively. In contrast, 44% (7 out of 16) of oyster samples and 30% (14 out of 47) of water samples from Alabama were positive. At both sites, there was greater sample-to-sample variability in pathogenic V. parahaemolyticus densities than in total V. parahaemolyticus densities. These data suggest that, although total V. parahaemolyticus densities may be very informative, there is greater uncertainty when total V. parahaemolyticus densities are used to predict the risk of infection by pathogenic V. parahaemolyticus than previously recognized. PMID:17921270

Variability of total and pathogenic Vibrio parahaemolyticus densities in northern Gulf of Mexico water and oysters.

PubMed

Zimmerman, A M; DePaola, A; Bowers, J C; Krantz, J A; Nordstrom, J L; Johnson, C N; Grimes, D J

2007-12-01

Vibrio parahaemolyticus is indigenous to coastal environments and a frequent cause of seafood-borne gastroenteritis in the United States, primarily due to raw-oyster consumption. Previous seasonal-cycle studies of V. parahaemolyticus have identified water temperature as the strongest environmental predictor. Salinity has also been identified, although it is evident that its effect on annual variation is not as pronounced. The effects of other environmental factors, both with respect to the seasonal cycle and intraseasonal variation, are uncertain. This study investigated intraseasonal variations of densities of total and pathogenic V. parahaemolyticus organisms in oysters and overlying waters during the summer of 2004 at two sites in the northern Gulf of Mexico. Regression analyses indicated significant associations (P < 0.001) between total V. parahaemolyticus densities and salinity, as well as turbidity in water and in oysters at the Mississippi site but not at the Alabama site. Pathogenic V. parahaemolyticus organisms in Mississippi oyster and water samples were detected in 56% (9 out of 16) and 78% (43 out of 55) of samples, respectively. In contrast, 44% (7 out of 16) of oyster samples and 30% (14 out of 47) of water samples from Alabama were positive. At both sites, there was greater sample-to-sample variability in pathogenic V. parahaemolyticus densities than in total V. parahaemolyticus densities. These data suggest that, although total V. parahaemolyticus densities may be very informative, there is greater uncertainty when total V. parahaemolyticus densities are used to predict the risk of infection by pathogenic V. parahaemolyticus than previously recognized.

The Association of Socio-Demographic Status, Lifestyle Factors and Dietary Patterns with Total Urinary Phthalates in Australian Men

PubMed Central

Bai, Peter Y.; Wittert, Gary A.; Taylor, Anne W.; Martin, Sean A.; Milne, Robert W.; Shi, Zumin

2015-01-01

Objective To investigate the associations between socio-demographic status, lifestyle factors, dietary patterns and urinary total phthalate concentration in a cohort of South Australian men. Method We randomly selected 1527 males aged 39 to 84 from wave two of the Men Androgen Inflammation Lifestyle Environment and Stress (MAILES) study. Total phthalate concentration was examined in fasting morning urine samples. Socio-demographic and lifestyle factors were assessed by questionnaire. Food intake was assessed by food frequency questionnaire (FFQ). Dietary patterns were constructed using factor analysis. Results Total phthalates were detected in 99.6% of the urine samples. The overall geometric mean (95% CI) of total phthalate concentration was 112.4 (107.5–117.5) ng/mL. The least square geometric means (LSGMs) of total phthalate concentration were significantly higher among people who were obese (127.8 ng/mL), consuming less than two serves fruit per day (125.7 ng/mL) and drinking more than one can (375mL) of carbonated soft drink per day (131.9 ng/mL). Two dietary patterns were identified: a prudent dietary pattern and a western dietary pattern. Both the western dietary pattern (p = 0.002) and multiple lifestyle risk factors including smoking, obesity, insufficient physical activity and the highest quartile of the western dietary pattern (p<0.001), were positively associated with total phthalate levels. There was no significant relationship between total phthalate concentration and socio-demographic status. Conclusion Phthalate exposure is ubiquitous and positively associated with lifestyle risk factors in urban dwelling Australian men. PMID:25875472

Monitoring of Foodborne Pathogens in Raw Cow Milk in Tuscany

PubMed Central

D’Alonzo, Alessia; Senese, Matteo; Fabbri, Ilaria; Cirri, Cristina; Milioni, Carla; Valenza, Valeria; Tolli, Rita; Campeis, Francesca; Fischetti, Roberto

2014-01-01

Raw milk consumption in Italy has increased over the last few years and although raw milk is characterised by cold chain, short shelf-life and the duty of boiling before domestic consumption, it is still considered a hazard. From 2010 to 2013 a monitoring survey of raw milk sold through vending machines was carried out to investigate the occurrence of several foodborne pathogens stipulated in the national legal requirements, i.e. Listeria monocytogenes, Campylobacter spp., Salmonella spp., Escherichia coli O:157 and coagulase-positive Staphylococci. A total of 127 raw milk samples were collected from 19 dairy herds in Tuscany Region, Italy. In addition, the milk samples were tested for the presence and count of Yersinia genus. Results shown that only one sample was positive for non verocytotoxin-producing E. coli O:157, whereas a total of 38 samples (29.9%) were postive for Yersinia genus; of the total 39 isolated bacteria, 23.6% were Y. enterocolitica, 2.4% Y. kristenseni and 4.7% Y. frederiksenii. None isolate was enteropathogenic; serotypes O:5 and O:8 were found in 16.6 and 13.3% of the isolates respectively, whereas none of the serotypes tested was detected in 70% of the isolates. The most probable number method revealed a count value between 0.03 and 24 MPN/mL. Based on these data a general assurance on health safety of raw milk produced and sold in Tuscany could be assessed. PMID:27800320

Evaluation of mercury pollution in cultivated and wild plants from two small communities of the Tapajós gold mining reserve, Pará State, Brazil.

PubMed

Egler, Silvia G; Rodrigues-Filho, Saulo; Villas-Bôas, Roberto C; Beinhoff, Christian

2006-09-01

This study examines the total Hg contamination in soil and sediments, and the correlation between the total Hg concentration in soil and vegetables in two small scale gold mining areas, São Chico and Creporizinho, in the State of Para, Brazilian Amazon. Total Hg values for soil samples for both study areas are higher than region background values (ca. 0.15 mg/kg). At São Chico, mean values in soils samples are higher than at Creporizinho, but without significant differences at alpha<0.05 level. São Chico's aboveground produce samples possess significantly higher values for total Hg levels than samples from Creporizinho. Creporizinho's soil-root produce regression model were significant, and the slope negative. Creporizinho's soil-aboveground and root wild plants regression models were also significant, and the slopes positives. Although, aboveground:root ratios were >1 in all of São Chico's produce samples, soil-plant parts regression were not significant, and Hg uptake probably occurs through stomata by atmospheric mercury deposition. Wild plants aboveground:root ratios were <1 at both study areas, and soil-plant parts regressions were significant in samples of Creporizinho, suggesting that they function as an excluder. The average total contents of Hg in edible parts of produces were close to FAO/WHO/JECFA PTWI values in São Chico area, and much lower in Creporizinho. However, Hg inorganic small gastrointestinal absorption reduces its adverse health effects.

TPH detection in groundwater: Identification and elimination of positive interferences

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zemo, D.A.; Synowiec, K.A.

1996-01-01

Groundwater assessment programs frequently require total petroleum hydrocarbon (TPH) analyses (Methods 8015M and 418.1). TPH analyses are often unreliable indicators of water quality because these methods are not constituent-specific and are vulnerable to significant sources of positive interferences. These positive interferences include: (a) non-dissolved petroleum constituents; (b) soluble, non-petroleum hydrocarbons (e.g., biodegradation products); and (c) turbidity, commonly introduced into water samples during sample collection. In this paper, we show that the portion of a TPH concentration not directly the result of water-soluble petroleum constituents can be attributed solely to these positive interferences. To demonstrate the impact of these interferences, wemore » conducted a field experiment at a site affected by degraded crude oil. Although TPH was consistently detected in groundwater samples, BTEX was not detected. PNAs were not detected, except for very low concentrations of fluorene (<5 ug/1). Filtering and silica gel cleanup steps were added to sampling and analyses to remove particulates and biogenic by-products. Results showed that filtering lowered the Method 8015M concentrations and reduced the Method 418.1 concentrations to non-detectable. Silica gel cleanup reduced the Method 8015M concentrations to non-detectable. We conclude from this study that the TPH results from groundwater samples are artifacts of positive interferences caused by both particulates and biogenic materials and do not represent dissolved-phase petroleum constituents.« less

TPH detection in groundwater: Identification and elimination of positive interferences

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zemo, D.A.; Synowiec, K.A.

1996-12-31

Groundwater assessment programs frequently require total petroleum hydrocarbon (TPH) analyses (Methods 8015M and 418.1). TPH analyses are often unreliable indicators of water quality because these methods are not constituent-specific and are vulnerable to significant sources of positive interferences. These positive interferences include: (a) non-dissolved petroleum constituents; (b) soluble, non-petroleum hydrocarbons (e.g., biodegradation products); and (c) turbidity, commonly introduced into water samples during sample collection. In this paper, we show that the portion of a TPH concentration not directly the result of water-soluble petroleum constituents can be attributed solely to these positive interferences. To demonstrate the impact of these interferences, wemore » conducted a field experiment at a site affected by degraded crude oil. Although TPH was consistently detected in groundwater samples, BTEX was not detected. PNAs were not detected, except for very low concentrations of fluorene (<5 ug/1). Filtering and silica gel cleanup steps were added to sampling and analyses to remove particulates and biogenic by-products. Results showed that filtering lowered the Method 8015M concentrations and reduced the Method 418.1 concentrations to non-detectable. Silica gel cleanup reduced the Method 8015M concentrations to non-detectable. We conclude from this study that the TPH results from groundwater samples are artifacts of positive interferences caused by both particulates and biogenic materials and do not represent dissolved-phase petroleum constituents.« less

Application of drug testing using exhaled breath for compliance monitoring of drug addicts in treatment.

PubMed

Carlsson, Sten; Olsson, Robert; Lindkvist, Irene; Beck, Olof

2015-04-01

Exhaled breath has recently been identified as a possible matrix for drug testing. This study explored the potential of this new method for compliance monitoring of patients being treated for dependence disorders. Outpatients in treatment programs were recruited for this study. Urine was collected as part of clinical routine and a breath sample was collected in parallel together with a questionnaire about their views of the testing procedure. Urine was analyzed for amphetamines, benzodiazepines, cannabis, cocaine, buprenorphine, methadone and opiates using CEDIA immunochemical screening and mass spectrometry confirmation. The exhaled breath was collected using the SensAbues device and analyzed by mass spectrometry for amphetamine, methamphetamine, diazepam, oxazepam, tetrahydrocannabinol, cocaine, benzoylecgonine, buprenorphine, methadone, morphine, codeine and 6-acetylmorphine. A total of 122 cases with parallel urine and breath samples were collected; 34 of these were negative both in urine and breath. Out of 88 cases with positive urine samples 51 (58%) were also positive in breath. Among the patients on methadone treatment, all were positive for methadone in urine and 83% were positive in breath. Among patients in treatment with buprenorphine, 92% were positive in urine and among those 80% were also positive in breath. The questionnaire response documented that in general, patients accepted drug testing well and that the breath sampling procedure was preferred. Compliance testing for the intake of prescribed and unprescribed drugs among patients in treatment for dependence disorders using the exhaled breath sampling technique is a viable method and deserves future attention.

[Incidence of Campylobacter spp. and Salmonella spp. in raw and roasted chicken in Guadalajara, Mexico].

PubMed

Castillo-Ayala, A; Salas-Ubiarco, M G; Márquez-Padilla, M L; Osorio-Hernández, M D

1993-01-01

The presence of Campylobacter spp. and Salmonella was studied in 70 samples of fresh retail chicken pieces and in 40 samples of roast chicken. Total plate count was performed in every sample as well. Most of the samples of fresh chicken yielded total plate counts > 10(8)/piece (thigh), while in roast chicken these counts ranged from 10(3) to 10(5)/piece (leg and thigh). Campylobacter was isolated from 33% of fresh chicken and from no sample of roast chicken. Salmonella was isolated from 69% of fresh chicken and 2.5% of roast chicken. There was no relationship between total plate counts in fresh chicken and isolation of either Campylobacter or Salmonella. Sixty percent of the Salmonella isolates belonged to serotype S. anatum, and about 50% of the isolates of Campylobacter were identified as being C. coli. The only Salmonella-positive sample of roast chicken yielded three serotypes: S. give, S. muenster, and S. manhattan. Presence of Campylobacter and Salmonella in chicken is of concern, due to the risk of spreading from the raw food to other cooked foods. The isolation of pathogens from roast chicken indicates mishandling during processing and/or storage of the product.

Minimally processed vegetable salads: microbial quality evaluation.

PubMed

Fröder, Hans; Martins, Cecília Geraldes; De Souza, Katia Leani Oliveira; Landgraf, Mariza; Franco, Bernadette D G M; Destro, Maria Teresa

2007-05-01

The increasing demand for fresh fruits and vegetables and for convenience foods is causing an expansion of the market share for minimally processed vegetables. Among the more common pathogenic microorganisms that can be transmitted to humans by these products are Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella. The aim of this study was to evaluate the microbial quality of a selection of minimally processed vegetables. A total of 181 samples of minimally processed leafy salads were collected from retailers in the city of Sao Paulo, Brazil. Counts of total coliforms, fecal coliforms, Enterobacteriaceae, psychrotrophic microorganisms, and Salmonella were conducted for 133 samples. L. monocytogenes was assessed in 181 samples using the BAX System and by plating the enrichment broth onto Palcam and Oxford agars. Suspected Listeria colonies were submitted to classical biochemical tests. Populations of psychrotrophic microorganisms >10(6) CFU/g were found in 51% of the 133 samples, and Enterobacteriaceae populations between 10(5) and 106 CFU/g were found in 42% of the samples. Fecal coliform concentrations higher than 10(2) CFU/g (Brazilian standard) were found in 97 (73%) of the samples, and Salmonella was detected in 4 (3%) of the samples. Two of the Salmonella-positive samples had <10(2) CFU/g concentrations of fecal coliforms. L. monocytogenes was detected in only 1 (0.6%) of the 181 samples examined. This positive sample was simultaneously detected by both methods. The other Listeria species identified by plating were L. welshimeri (one sample of curly lettuce) and L. innocua (2 samples of watercress). The results indicate that minimally processed vegetables had poor microbiological quality, and these products could be a vehicle for pathogens such as Salmonella and L. monocytogenes.

Monitoring baseline suspended sediment in forested basins: the effects of sampling on suspended sediment rating curves

Treesearch

Robert B. Thomas

1988-01-01

Abstract - Rating curves are widely used for directly assessing changes in the suspended sediment delivery process and indirectly for estimating total yields. Four sampling methods were simulated-over a 31-day record of suspended sediment from the North Fork of the Mad River near Korbel, California. The position and size of the four groups of plotted slope/intercept...

Comparison of two real-time PCR assays for the detection of malaria parasites from hemolytic blood samples - Short communication.

PubMed

Hagen, Ralf Matthias; Hinz, Rebecca; Tannich, Egbert; Frickmann, Hagen

2015-06-01

We compared the performance of an in-house and a commercial malaria polymerase chain reaction (PCR) assay using freeze-thawed hemolytic blood samples. A total of 116 freeze-thawed ethylenediamine tetraacetic acid (EDTA) blood samples of patients with suspicion of malaria were analyzed by an in-house as well as by a commercially available real-time PCR. Concordant malaria negative PCR results were reported for 39 samples and malaria-positive PCR results for 67 samples. The in-house assay further detected one case of Plasmodium falciparum infection, which was negative in the commercial assay as well as five cases of P. falciparum malaria and three cases of Plasmodium vivax malaria, which showed sample inhibition in the commercial assay. The commercial malaria assay was positive in spite of a negative in-house PCR result in one case. In all concordant results, cycle threshold values of P. falciparum-positive samples were lower in the commercial PCR than in the in-house assay. Although Ct values of the commercial PCR kit suggest higher sensitivity in case of concordant results, it is prone to inhibition if it is applied to hemolytic freeze-thawed blood samples. The number of misidentifications was, however, identical for both real-time PCR assays.

[Detection of herpes virus and human enterovirus in pathology samples using low-density arrays].

PubMed

Del Carmen Martínez, Sofía; Gervás Ríos, Ruth; Franco Rodríguez, Yoana; González Velasco, Cristina; Cruz Sánchez, Miguel Ángel; Abad Hernández, María Del Mar

Despite the frequency of infections with herpesviridae family, only eight subtypes affect humans (Herpex Simplex Virus types 1 and 2, Varicella Zoster Virus, Epstein-Barr Virus, Citomegalovirus and Human Herpes Virus types 6, 7 and 8). Amongst enteroviruses infections, the most important are Poliovirus, Coxackievirus and Echovirus. Symptoms can vary from mild to severe and early diagnosis is of upmost importance. Nowadays, low-density arrays can detect different types of viruses in a single assay using DNA extracted from biological samples. We analyzed 70 samples of formalin-fixed and paraffin-embedded tissue, searching for viruses (HSV-1, HSV-2, VZV, CMV, EBV, HHV-6, HHV-7 y HHV-8, Poliovirus, Echovirus and Coxsackievirus) using the kit CLART ® ENTHERPEX. Out of the total of 70 samples, 29 were positive for viral infection (41.43%), and only 4 of them showed cytopathic effect (100% correlation between histology and the test). 47.6% of GVHD samples were positive for virus; 68.75% of IBD analyzed showed positivity for viral infection; in colitis with ulcers (neither GVHD nor IBD), the test was positive in 50% of the samples and was also positive in 50% of ischemic lesions. The high sensitivity of the technique makes it a useful tool for the pathologist in addition to conventional histology-based diagnosis, as a viral infection may affect treatment. Copyright © 2016 Sociedad Española de Anatomía Patológica. Publicado por Elsevier España, S.L.U. All rights reserved.

Identification and characterization of Dichelobacter nodosus serogroup H from ovine footrot in India.

PubMed

Vinod Kumar, N; Sreenivasulu, D; Karthik, A

2016-08-01

A total of 56 foot swabs were collected from inter digital spaces of sheep with footrot lesions were screened for 16 rRNA of Dichelobacter nodosus by PCR. Out of the 56 samples, 38(67.85%) were found to be positive. All the positive samples were subjected to multiplex PCR targeting fimA gene for identification of serogroups of D. nodosus. Serogroup H was found along with serogroup B in 12 (55.26%) samples and with serogroup I in 8 (22.2%) samples. The serogroup H was identified for the first time from the Indian subcontinent. The phylogenetic analysis of the present sequence with the available serogroup H sequences of GenBank revealed to be in close association with the serotype H1. Copyright © 2016 Elsevier Ltd. All rights reserved.

Short communication: Evaluation of sampling socks for detection of Mycobacterium avium ssp. paratuberculosis on dairy farms.

PubMed

Wolf, R; Orsel, K; De Buck, J; Kanevets, U; Barkema, H W

2016-04-01

Mycobacterium avium ssp. paratuberculosis (MAP) causes Johne's disease, a production-limiting disease in cattle. Detection of infected herds is often done using environmental samples (ES) of manure, which are collected in cattle pens and manure storage areas. Disadvantages of the method are that sample accuracy is affected by cattle housing and type of manure storage area. Furthermore, some sampling locations (e.g., manure lagoons) are frequently not readily accessible. However, sampling socks (SO), as used for Salmonella spp. testing in chicken flocks, might be an easy to use and accurate alternative to ES. The objective of the study was to assess accuracy of SO for detection of MAP in dairy herds. At each of 102 participating herds, 6 ES and 2 SO were collected. In total, 45 herds had only negative samples in both methods and 29 herds had ≥1 positive ES and ≥1 positive SO. Furthermore, 27 herds with ≥1 positive ES had no positive SO, and 1 herd with no positive ES had 1 positive SO. Bayesian simulation with informative priors on sensitivity of ES and MAP herd prevalence provided a posterior sensitivity for SO of 43.5% (95% probability interval=33-58), and 78.5% (95% probability interval=62-93) for ES. Although SO were easy to use, accuracy was lower than for ES. Therefore, with improvements in the sampling protocol (e.g., more SO per farm and more frequent herd visits), as well as improvements in the laboratory protocol, perhaps SO would be a useful alternative for ES. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Analysis of beta-lactam antibiotics in incurred raw milk by rapid test methods and liquid chromatography coupled with electrospray ionization tandem mass spectrometry.

PubMed

Riediker, S; Diserens, J M; Stadler, R H

2001-09-01

A recently developed confirmatory LC-MS method has been applied to the quantification of five major beta-lactam antibiotics in suspect raw bovine milk samples that gave a positive response with receptor-based (BetaStar) and rapid microbial inhibitory screen tests (Delvotest SP). In total, 18 presumptive positive raw milk samples were reanalyzed; 16 samples showed traces of antibiotic residues that could be identified and quantified by the LC-MS method, ranging from the limits of confirmation up to 38 microg/kg. Of the positive samples, only five (approximately 30%) were found to be violative of EU maximum residue limits. The most frequently detected antibiotic residues were cloxacillin and penicillin G, the former often in combination with amoxicillin or ampicillin. This study compares the results obtained by the three methods on identical samples and addresses how these relate to certain criteria such as sensitivity and selectivity. Furthermore, the limitations of the LC-MS method and the potential impact of the presence of frequently more than one residue in the same milk sample on the response of the rapid test methods are discussed.

Total mercury and methylmercury in fish fillets, water, and bed sediments from selected streams in the Delaware River basin, New Jersery, New York, and Pennsylvania, 1998-2001

USGS Publications Warehouse

Brightbill, Robin A.; Riva-Murray, Karen; Bilger, Michael D.; Byrnes, John D.

2004-01-01

Within the Delaware River Basin, fish-tissue samples were analyzed for total mercury (tHg). Water and bed-sediment samples were analyzed for tHg and methylmercury (MeHg), and methylation efficiencies were calculated. This study was part of a National Mercury Pilot Program conducted by the U.S. Geological Survey (USGS). The Delaware River Basin was chosen because it is part of the USGS National Water-Quality Assessment Program that integrates physical, chemical, and biological sampling efforts to determine status and trends in surface-water and ground-water resources. Of the 35 sites in the study, 31 were sampled for fish. The species sampled at these sites include smallmouth bass (Micropterus dolomieu), the target species, and where smallmouth bass could not be collected, brown trout (Salmo trutta), chain pickerel (Esox niger), largemouth bass (Micropterus salmoides), and rock bass (Ambloplites rupestris). There were a total of 32 fish samples; 7 of these exceeded the 0.3 ?g/g (micrograms per gram) wet-weight mercury (Hg) concentration set for human health by the U.S. Environmental Protection Agency and 27 of these exceeded the U.S. Fish and Wildlife Service criteria of 0.1 ?g/g wet weight for the protection of fish-eating birds and wildlife. Basinwide analysis of Hg in fish, water, and bed sediment showed tHg concentration in fillets correlated positively with population density, urban land cover, and impervious land surface. Negative correlations included wetland land cover, septic density, elevation, and latitude. Smallmouth bass from the urban sites had a higher median concentration of tHg than fish from agricultural, low intensity-agricultural, or forested sites. Concentrations of tHg and MeHg in water were higher in samples from the more urbanized areas of the basin and were positively correlated with urbanization and negatively correlated with forested land cover. Methylation efficiency of water was negatively correlated with urbanization. Bed-sediment patterns were similar to those observed in water. Concentrations of tHg were higher in samples from the urbanized areas. In the more forested areas, MeHg concentrations were higher than in other land-use areas. Concentrations of tHg in bed sediment were positively correlated with urbanization factors (population, urban land cover, and impervious land surface) and negatively correlated with forested land cover and elevation. Forested land cover and latitude were positively correlated with concentrations of MeHg. The methylation efficiency was higher in samples from the forested areas and was negatively correlated with urbanization. Analyses within land-use groups showed that tHg concentrations in fish fillets from the urban sites were positively correlated with forested land cover and wetland cover. Urbanization factors within the agricultural group were positively correlated with tHg in fish; concentrations of tHg in fish from sites in the low intensity-agricultural group were negatively correlated with urbanization factors. Within the agricultural land-use group, tHg concentrations in water were negatively correlated with septic density, and MeHg concentrations were negatively correlated with elevation. In the forested and low intensity-agricultural groups, MeHg in water was negatively correlated with forested and agricultural land cover. Methylation efficiency in water also was negatively correlated with forested land cover but positively correlated with agricultural land cover. Bed sediment concentrations of tHg in the forested and low-agricultural groups were positively correlated with agricultural land cover and negatively correlated with forested land cover. Concentrations of MeHg in bed sediment were positively correlated with septic density and drainage area and negatively correlated with forested land cover. Methylation efficiency was negatively correlated with population density, a

A Cross-Reactivity of Fenofibric Acid With MDMA DRI Assay.

PubMed

Bugier, Sarah; Garcia-Hejl, Carine; Vest, Philippe; Plantamura, Julie; Chianea, Denis; Renard, Christophe

2016-09-01

Within the framework of routine fitness examinations, French Air Force military crew underwent urine testing for 3,4 methylenedioxymetamphetamine (MDMA [ecstasy]). The cross-reactivity of a dyslipidemic drug, fenofibrate, with an MDMA immunoassay was studied and confirmed on a large population sample. A 3-year retrospective study was performed on the MDMA DRI Ecstasy Assay on the Unicel DXC 600. In the event of positive test result, a confirmatory testing was carried out by gas chromatography/mass spectrometry (GC/MS) to establish the presence of MDMA. When analysis by GC/MS did not confirm the presence of MDMA, a false-positive result was suspected and the samples were analyzed by high-performance liquid chromatography-mass spectrometry to identify a potential interfering substance. A total of 15,169 urine samples, from 7,803 patients, were tested for 3 years. Of the tested samples, 22 (0.15%) were positive by DRI Ecstasy Assay. None of them were positive by GC/MS. A cross-reactivity of fenofibrate's metabolite with MDMA using this assay was systematically found. Fenofibrate's interference with MDMA immunoassay was confirmed. Fenofibrate being widely prescribed, physicians had to be alerted that this treatment could lead to false-positive results. Reprint & Copyright © 2016 Association of Military Surgeons of the U.S.

Smoking-Related Knowledge, Attitudes, Behaviors, Smoking Cessation Idea and Education Level among Young Adult Male Smokers in Chongqing, China

PubMed Central

Xu, Xianglong; Liu, Lingli; Sharma, Manoj; Zhao, Yong

2015-01-01

Introduction: In 2012 in China, 52.9% of men were reported to smoke while only 2.4% of women smoked. This study explored the smoking-related Knowledge, Attitudes and Practices (KAP) among young adult male smokers. Methods: A cross-sectional study was conducted in four municipal areas of Chongqing using a questionnaire administered to 536 natives young male smokers aged 18–45 years old. Results: The total score of smoking cognition, the total score of smoking attitude and the total score of positive behavior to quit smoking was significantly different among the three groups by education. Besides, 30.97% of male smokers never seriously thought about quitting smoking. Logistic regression analysis found smoking-related knowledge, attitudes, behaviors and sociodemographic factors affect having smoking cessation idea. But no statistically significant correlation was observed between smoking cognition and positive behavior to quit smoking in a sample of higher education. No statistically significant correlation was observed between smoking cognition and positive behavior to quit smoking (Pearson correlation coefficient = 0.03012, p = 0.6811), and also no statistically significant correlation was observed between smoking cognition and positive behavior to quit smoking (Pearson correlation coefficient = 0.08869, p = 0.2364)  in the sample of higher education young adult males Conclusions: Young adult males with higher education have a better knowledge of smoking hazards and a more positive attitude toward smoking, however, this knowledge and attitude do not necessarily translate into health behavioral outcomes such as not smoking. Overall the present findings indicate that no statistically significant correlation between the education level and quitting smoking idea exists among young adult male smokers in China. This survey gives a snapshot of the impact of education on smoking-related KAP among young adults male smokers. PMID:25689992

Smoking-related knowledge, attitudes, behaviors, smoking cessation idea and education level among young adult male smokers in Chongqing, China.

PubMed

Xu, Xianglong; Liu, Lingli; Sharma, Manoj; Zhao, Yong

2015-02-16

In 2012 in China, 52.9% of men were reported to smoke while only 2.4% of women smoked. This study explored the smoking-related Knowledge, Attitudes and Practices (KAP) among young adult male smokers. A cross-sectional study was conducted in four municipal areas of Chongqing using a questionnaire administered to 536 natives young male smokers aged 18-45 years old. The total score of smoking cognition, the total score of smoking attitude and the total score of positive behavior to quit smoking was significantly different among the three groups by education. Besides, 30.97% of male smokers never seriously thought about quitting smoking. Logistic regression analysis found smoking-related knowledge, attitudes, behaviors and sociodemographic factors affect having smoking cessation idea. But no statistically significant correlation was observed between smoking cognition and positive behavior to quit smoking in a sample of higher education. No statistically significant correlation was observed between smoking cognition and positive behavior to quit smoking (Pearson correlation coefficient = 0.03012, p = 0.6811), and also no statistically significant correlation was observed between smoking cognition and positive behavior to quit smoking (Pearson correlation coefficient = 0.08869, p = 0.2364) in the sample of higher education young adult males Young adult males with higher education have a better knowledge of smoking hazards and a more positive attitude toward smoking, however, this knowledge and attitude do not necessarily translate into health behavioral outcomes such as not smoking. Overall the present findings indicate that no statistically significant correlation between the education level and quitting smoking idea exists among young adult male smokers in China. This survey gives a snapshot of the impact of education on smoking-related KAP among young adults male smokers.

Feasibility of Estimating Constituent Concentrations and Loads Based on Data Recorded by Acoustic Instrumentation

USGS Publications Warehouse

Lietz, A.C.

2002-01-01

The acoustic Doppler current profiler (ADCP) and acoustic Doppler velocity meter (ADVM) were used to estimate constituent concentrations and loads at a sampling site along the Hendry-Collier County boundary in southwestern Florida. The sampling site is strategically placed within a highly managed canal system that exhibits low and rapidly changing water conditions. With the ADCP and ADVM, flow can be gaged more accurately rather than by conventional field-data collection methods. An ADVM velocity rating relates measured velocity determined by the ADCP (dependent variable) with the ADVM velocity (independent variable) by means of regression analysis techniques. The coefficient of determination (R2) for this rating is 0.99 at the sampling site. Concentrations and loads of total phosphorus, total Kjeldahl nitrogen, and total nitrogen (dependent variables) were related to instantaneous discharge, acoustic backscatter, stage, or water temperature (independent variables) recorded at the time of sampling. Only positive discharges were used for this analysis. Discharges less than 100 cubic feet per second generally are considered inaccurate (probably as a result of acoustic ray bending and vertical temperature gradients in the water column). Of the concentration models, only total phosphorus was statistically significant at the 95-percent confidence level (p-value less than 0.05). Total phosphorus had an adjusted R2 of 0.93, indicating most of the variation in the concentration can be explained by the discharge. All of the load models for total phosphorus, total Kjeldahl nitrogen, and total nitrogen were statistically significant. Most of the variation in load can be explained by the discharge as reflected in the adjusted R2 for total phosphorus (0.98), total Kjeldahl nitrogen (0.99), and total nitrogen (0.99).

Grouped fuzzy SVM with EM-based partition of sample space for clustered microcalcification detection.

PubMed

Wang, Huiya; Feng, Jun; Wang, Hongyu

2017-07-20

Detection of clustered microcalcification (MC) from mammograms plays essential roles in computer-aided diagnosis for early stage breast cancer. To tackle problems associated with the diversity of data structures of MC lesions and the variability of normal breast tissues, multi-pattern sample space learning is required. In this paper, a novel grouped fuzzy Support Vector Machine (SVM) algorithm with sample space partition based on Expectation-Maximization (EM) (called G-FSVM) is proposed for clustered MC detection. The diversified pattern of training data is partitioned into several groups based on EM algorithm. Then a series of fuzzy SVM are integrated for classification with each group of samples from the MC lesions and normal breast tissues. From DDSM database, a total of 1,064 suspicious regions are selected from 239 mammography, and the measurement of Accuracy, True Positive Rate (TPR), False Positive Rate (FPR) and EVL = TPR* 1-FPR are 0.82, 0.78, 0.14 and 0.72, respectively. The proposed method incorporates the merits of fuzzy SVM and multi-pattern sample space learning, decomposing the MC detection problem into serial simple two-class classification. Experimental results from synthetic data and DDSM database demonstrate that our integrated classification framework reduces the false positive rate significantly while maintaining the true positive rate.

Prevalence of Chlamydia psittaci and Other Chlamydia Species in Wild Birds in Poland.

PubMed

Krawiec, Marta; Piasecki, Tomasz; Wieliczko, Alina

2015-11-01

Avian chlamydiosis is a zoonotic disease occurring in humans, poultry, and exotic birds. It has been suggested that some wild bird species play an important role as reservoirs for Chlamydia, especially Chlamydia psittaci. Whereas C. psittaci is the predominant chlamydial agent in birds, in the present study we have determined the prevalence of different species of Chlamydia among selected wild bird species in Poland using a rapid and sensitive real-time PCR method. In total, 369 free-living birds from 35 bird species and 15 orders were examined. Samples from 27 birds (7.3%) were positive for chlamydial DNA in the PCR; 22 positive samples (81.5%) belonged to C. psittaci, three to Chlamydia trachomatis (11.1%), and two (7.4%) classified only to the genus Chlamydia. Most of C. psittaci-positive samples belonged to five orders: Anseriformes, Columbiformes, Gruiformes, Phasianiformes, and Passeriformes. All C. trachomatis samples were obtained from Eurasian coots (Gruiformes). Two Chlamydia-positive samples not classified to any Chlamydia species were obtained from a common wood pigeon (Columbiformes) and a common buzzard (Accipitriformes). Detection of C. psittaci and C. trachomatis in free-living bird populations force to think on significance of birds as reservoir of varied Chlamydia species and their epidemiological importance.

Detection of Toxoplasma gondii in raw caprine, ovine, buffalo, bovine, and camel milk using cell cultivation, cat bioassay, capture ELISA, and PCR methods in Iran.

PubMed

Dehkordi, Farhad Safarpoor; Borujeni, Mohammad Reza Haghighi; Rahimi, Ebrahim; Abdizadeh, Rahman

2013-02-01

This study was conducted to determine the presence of Toxoplasma gondii in animal milk samples in Iran. From a total of 395 dairy herds in three provinces of Iran, 66 bovine, 58 ovine, 54 caprine, 33 buffalo, and 30 camel herds were studied, and from these parts of Iran, 200 bovine, 185 ovine, 180 caprine, 164 buffalo, and 160 camel milk samples were collected from various seasons. Samples were tested for Toxoplasma gondii by cell line culture, enzyme-linked immunosorbent assay (ELISA), and polymerase chain reaction (PCR) technique. Only the results of cell line cultivation were confirmed by bioassay in cat. Results indicated that all herds were infected with Toxoplasma gondii. The culture method showed that 51 out of 889 milk samples (5.73%) were positive for Toxoplasma gondii, and all 51 positive culture results were positive with bioassay in cat. The Fars province had the highest prevalence of Toxoplasma gondii (6.84%). The ELISA test showed that 41 milk samples (4.61%) were positive for the presence of Toxoplasma gondii, while the PCR showed that 46 milk samples were positive for Toxoplasma gondii. The results showed higher sensitivity of PCR and higher specificity of ELISA. Caprine had the highest (10%) and camel had the lowest (3.12%) prevalence rate of parasite. The summer season had the highest (76.47%) but winter (3.92) had the lowest incidence of Toxoplasma gondii. This study is the first prevalence report of direct detection of Toxoplasma gondii in animal milk samples in Iran.

Detection of quinolones in commercial eggs obtained from farms in the Espaíllat Province in the Dominican Republic.

PubMed

Moscoso, S; de los Santos, F Solís; Andino, A G; Diaz-Sanchez, Sandra; Hanning, I

2015-01-01

Previously, we reported the use of quinolones in broiler chickens resulted in residues in retail poultry meat obtained from nine districts in the Santiago Province of the Dominican Republic. Residues in poultry products are a concern due to consumer allergies and the potential to develop antibiotic-resistant bacteria. Given the use of quinolones in poultry production and our previous findings in poultry meat, the objective of this study was to evaluate the presence of quinolone residues in eggs. Samples were collected from 48 different farms located in three of the four municipalities (Moca, Cayetano Germosén, and Jamao) of the Espaíllat Province. Each farm was sampled three times between July and September for a total of 144 samples. Samples were evaluated qualitatively and quantitatively for quinolone residues using the Equinox test. Operation systems (cage or floor), seasonality, and location were considered along with egg-producer sizes that were defined as small scale, <30,000 eggs per day; medium scale, 30,000 to 60,000 eggs per day; or large scale, >60,000 eggs per day. From small-, medium-, and large-scale producers, 69, 50, and 40% of samples were positive for quinolone residues, respectively. A greater number of samples were positive (61%) in floor-laying hen producers compared with those using cages (40%). In the Jamao municipality, 67% of the samples were positive compared with Moca and Cayetano Germosén, where 56 and 25% of samples were positive, respectively. Sampling time had an effect on percent positives: samples collected in July, August, and September were 71, 19, and 63% positive, respectively. Overall, 51% of the samples obtained from eggs produced in the province of Espaíllat were positive for quinolone residues at levels higher than the maximum limits for edible tissue established by the regulatory agencies, including the European Union and U.S. Department of Agriculture. The results obtained from this research confirmed the presence of quinolone residue in eggs, which may present a health risk to some consumers.

An assessment of the microbiological quality and safety of raw drinking milk on retail sale in England.

PubMed

Willis, C; Jørgensen, F; Aird, H; Elviss, N; Fox, A; Jenkins, C; Fenelon, D; Sadler-Reeves, L; McLauchlin, J

2018-02-01

This study aimed to review the microbiological results for raw drinking milk (RDM) samples submitted to Public Health England laboratories between 2014 and 2016 in order to produce up-to-date data on the microbiological safety of RDM and inform future risk assessments on its sale. A total of 902 samples of RDM were collected from retail sale in England for microbiological examination. Overall, 454 of 770 samples (59·0%) taken for routine monitoring were of a satisfactory quality, whilst eight (1·0%) were 'unsatisfactory and potentially injurious to health' due to the presence of Shiga toxin-producing Escherichia coli, Campylobacter or elevated levels of Listeria monocytogenes or coagulase-positive staphylococci. In contrast, 16 of 114 (14·0%) of samples taken in follow-up to a previous unsatisfactory result and 5 of 18 (27·8%) of samples related to illness were potentially injurious. A total of 229 of 902 samples (25·4%) gave unsatisfactory results due to elevated aerobic colony counts and/or coliforms, whilst 139 of 902 samples (15·4%) were of borderline quality due to coagulase-positive staphylococci. Listeria monocytogenes was detected at levels of <100 CFU per ml in 66 of 902 samples (7·3%) and other Listeria species in 44 of 902 samples (4·9%). Pathogens and/or indicators of poor hygiene were present in almost half of samples examined. Cows' milk samples gave a significantly greater proportion of unsatisfactory results compared to milk from other species (i.e. goat, sheep, buffalo, camel). These results demonstrate the importance of maintaining strict controls on the production and sale of this product. © 2017 The Society for Applied Microbiology.

The Role of the Five Factor Model of Personality in the Perceptions of Negative and Positive Forms of Work-Nonwork Spillover: A Meta-Analytic Review

ERIC Educational Resources Information Center

Michel, Jesse S.; Clark, Malissa A.; Jaramillo, David

2011-01-01

The present meta-analysis examines the relationships between the Five Factor Model (FFM) of personality and negative and positive forms of work-nonwork spillover (e.g., work-family conflict and facilitation). Results, based on aggregated correlations drawn from 66 studies and 72 independent samples (Total N = 28,127), reveal that the FFM is…

Determination of chloramphenicol residues in commercial chicken eggs in the Federal Capital Territory, Abuja, Nigeria.

PubMed

Mbodi, Felix E; Nguku, P; Okolocha, E; Kabir, J

2014-01-01

The use of antibiotics in poultry can result in residues in eggs. The joint FAO/WHO committee recommended banning the use of chloramphenicol (CAP) in food animals due to its public health hazards of aplastic anaemia, leukaemia, allergy, antibacterial resistance and carcinogenicity. This paper determines the prevalence of CAP residues in chicken eggs and assesses the usage and awareness of its ban amongst poultry farmers in the Federal Capital Territory (FCT), Abuja, Nigeria. A cross-sectional survey of registered poultry farmers in FCT was conducted using questionnaires to determine CAP administration in poultry and awareness of its ban. Pooled egg samples were collected from each poultry farm surveyed and from randomly sampled government-owned markets in FCT. Source of eggs by state were identified by the marketer at the time of collection. Samples were analysed using an enzyme-linked immunosorbent assay (ELISA) technique for the presence of CAP, and prevalence was determined. Of 288 total pooled samples collected, 257 (89.2%) were from the markets and 31 (10.8%) were from poultry farms. A total of 20 (7%) pooled egg samples tested CAP-positive; market eggs originated from 15 (41%) states of the country. Of the market eggs, 16 (6.2%) pooled samples tested positive. Of eggs from poultry farms, four (12.9%) tested positive. Mean CAP concentrations in the positive samples ranged from 0.49 to 1.17 µg kg(-1) (parts per billion). CAP use amongst poultry farmers in FCT was 75.5%; awareness of the CAP ban was 26.3%. Though 66% of veterinarians were unaware of a CAP ban, they were more likely to be aware than other poultry farmers (odds ratio (OR) = 1.4). Farm managers who use CAP were more likely to be aware of CAP ban than the farm managers not using CAP (OR = 5.5; p = 0.04). Establishing a drug residue surveillance and control program and enforcement of CAP legislation/regulation is needful to educate and prohibit the widespread CAP use amongst Nigerian poultry farmers.

A 10-day vacancy period after cleaning and disinfection has no effect on the bacterial load in pig nursery units.

PubMed

Luyckx, K; Millet, S; Van Weyenberg, S; Herman, L; Heyndrickx, M; Dewulf, J; De Reu, K

2016-10-19

Biosecurity measures such as cleaning, disinfection and a vacancy period between production cycles on pig farms are essential to prevent disease outbreaks. No studies have tested the effect of a longer vacancy period on bacterial load in nursery units. The present study evaluated the effect of a 10-day vacancy period in pig nursery units on total aerobic flora, Enterococcus spp., Escherichia coli, faecal coliforms and methicillin resistant Staphylococcus aureus (MRSA). Three vacancy periods of 10 days were monitored, each time applied in 3 units. The microbiological load was measured before disinfection and at 1, 4, 7 and 10 days after disinfection. No significant decrease or increase in E. coli, faecal coliforms, MRSA and Enterococcus spp. was noticed. Total aerobic flora counts were the lowest on day 4 after disinfection (i.e. 4.07 log CFU/625 cm 2 ) (P < 0.05), but the difference with other sampling moments was limited (i.e. 0.6 log CFU/625 cm 2 ) and therefore negligible. Furthermore, this observation on day 4 was not confirmed for the other microbiological parameters. After disinfection, drinking nipples were still mostly contaminated with total aerobic flora (i.e. 5.32 log CFU/625 cm 2 ) and Enterococcus spp. (i.e. 95 % of the samples were positive) (P < 0.01); the feeding troughs were the cleanest location (total aerobic flora: 3.53 log CFU/625 cm 2 and Enterococcus spp.: 50 % positive samples) (P < 0.01). This study indicates that prolonging the vacancy period in nursery units to 10 days after disinfection with no extra biosecurity measures has no impact on the environmental load of total aerobic flora, E. coli, faecal coliforms, MRSA and Enterococcus spp..

Relation between selected well-construction characteristics and occurrence of bacteria in private household-supply wells, south-central and southeastern Pennsylvania

USGS Publications Warehouse

Zimmerman, Tammy M.; Zimmerman, Michele L.; Lindsey, Bruce D.

2001-01-01

Total coliform and Escherichia coli (E. coli) bacteria were analyzed in ground water sampled from 78 private household-supply wells as part of a study by the U.S. Geological Survey in cooperation with the Pennsylvania Department of Environmental Protection to evaluate the relation between well construction characteristics and the occurrence of bacteria in ground water. Sampling was done in eight counties in south-central and southeastern Pennsylvania from September 2000 to March 2001. All samples were collected from wells in close proximity to agricultural land-use areas.Total coliform bacteria were found in water from 62 percent (48 of 78) of the wells, and bacteria were just as likely to be found in sanitary wells (grouted/loose-fitting well cap or grouted/sanitary sealed well cap) as in nonsanitary wells (nongrouted/ loose-fitting well cap). The areas underlain by carbonate bedrock had the highest percentages of total coliform detected (about 75 percent). Nearly half of the samples collected in the areas underlain by noncarbonate bedrock also were found to have total coliform present. E. coli bacteria were found in water from 10 percent of the wells. Seventeen percent of the samples that were positive for total coliform also were positive for E. coli. The presence of E. coli bacteria was more likely in water from nonsanitary wells. Additionally, the presence of E. coli bacteria was more likely in ground water from wells underlain by carbonate bedrock. A further breakdown of the data into four groups on the basis of sanitary construction and bedrock type indicated the presence of E. coli was more likely in water from nonsanitary wells in areas underlain by carbonate bedrock.Statistical analysis of other well-construction characteristics that might relate to occurrence of bacteria showed that the presence of total coliform bacteria was related to the depth to water-bearing zone in both sanitary and nonsanitary wells in areas underlain by carbonate bedrock. Relations also are present between the presence of total coliform bacteria and casing length in nonsanitary wells in areas underlain by noncarbonate bedrock. Bacteria were found in wells both with and without insects observed on the underside of the well cap. Because of the small number of wells sampled that had sanitary sealed caps, it is uncertain whether installation of sanitary sealed well caps would reduce the incidence of bacteria in ground water from wells or if the presence of bacteria is because of a combination of well-construction characteristics or aquifer-wide contamination of limited or broad areal extent.

Seroprevalence of Crimean-Congo Hemorrhagic Fever Virus in Erzincan Province, Turkey, Relationship with Geographic Features and Risk Factors.

PubMed

Cikman, Aytekin; Aydin, Merve; Gulhan, Baris; Karakecili, Faruk; Kesik, Ozan Arif; Ozcicek, Adalet; Akin, Hicran; Kara, Murat

2016-03-01

To determine the seroprevalence and risk factors associated with Crimean-Congo hemorrhagic fever virus (CCHFV) in residents of Erzincan, Turkey. Although CCHFV is endemic in Erzincan, this is the first study to evaluate its seroprevalence in this region. This study included a total of 372 subjects, 174 of whom had been exposed to or bitten by ticks, 145 of whom worked with livestock, and 53 of whom resided in the city and did not have exposure to livestock. Data on CCHFV IgG and IgM antibodies were extracted from serum samples collected from all subjects using an ELISA. All samples were tested for CCHFV IgG and CCHFV IgM. Only IgM-positive samples were processed for detection of viral RNA through RT-PCR. Using seropositive cases only, we performed spatial analyses to evaluate correlations between seroprevalence and geographic location (i.e., proximity to rivers, altitude, and slope angle of land). In this study, 14.0% (52/322) of the total subjects were positive for CCHFV IgG. Seven of the individuals were positive both for CCHFV IgG and CCHFV IgM. Of these seven, only one sample tested positive for CCHFV RNA. Individuals who worked with livestock in the rural areas and had a history of tick exposure were statistically more likely to test positive for CCHFV IgG than individuals from the city and not exposed to ticks (p < 0.05). Seroprevalence was affected by geographic characteristics, including distance to rivers, altitude, and slope angle of land. We observed a high seroprevalence of CCHFV in Erzincan, which is similar to that observed in other endemic regions of Turkey. CCHFV seroprevalence rates are found to be quite high in the people who live in the sloping fields at certain heights and where there are a lot of rivers and streams.

Prevalence of avian influenza (H9N2) in commercial quail, partridge, and turkey farms in Iran, 2014-2015.

PubMed

Mehrabadi, Mohammad Hosein Fallah; Bahonar, Alireza; Mirzaei, Kamran; Molouki, Aidin; Ghalyanchilangeroudi, Arash; Ghafouri, Seyed Ali; Tehrani, Farshad; Lim, Swee Hua Erin

2018-03-01

Avian influenza virus (AIV) H9N2 subtype is endemic in Iran and causes substantial economic loss to the growing poultry industry within the country. In this study, a cross-sectional analysis was carried out to determine the sero-prevalence of H9N2 in several commercial farms between the years 2014 and 2015. The comparison of the mean of serum titers and the ratio of sero-positive birds between all units were analyzed using one-way ANOVA test. In 2014, a total of 77 farms (58 turkey farms, 14 quail farms, and 5 partridge farms) and 894 birds (682 turkeys, 154 quails, and 58 partridges) were sampled while in 2015, a total of 69 farms (54 turkey farms, 8 quail farms, and 7 partridge farms) and 856 birds (675 turkeys, 105 quails, and 76 partridges) were sampled. Of that, 52 of 77 sampled farms (67.5%) and 437 of 894 samples (48.9%) were positive for H9N2 in 2014 while. Forty-one of 69 farms (59.4%) and 307 of 856 sera (35.9%) were positive in 2015. Furthermore, the mean titer of partridge farms was significantly lower than that of turkey farms (p < 0.01) and the mean percentage of sero-positive turkey farms was significantly higher than partridge farms (p < 0.01) in 2014. In 2015, no significant difference was observed between the mean sera titer amongst farms and percentage of sero-positive birds (p > 0.05). Our results indicated that H9N2 is circulating in these farms. Since many more such farms are being established for operations, in addition to the threat of emergence and continuous reemergence of the disease in these farms, enhanced veterinary biosecurity measures on farms are required for mitigation.

Chemical contaminants in water and sediment near fish nesting sites in the Potomac River basin: determining potential exposures to smallmouth bass (Micropterus dolomieu).

PubMed

Kolpin, Dana W; Blazer, Vicki S; Gray, James L; Focazio, Michael J; Young, John A; Alvarez, David A; Iwanowicz, Luke R; Foreman, William T; Furlong, Edward T; Speiran, Gary K; Zaugg, Steven D; Hubbard, Laura E; Meyer, Michael T; Sandstrom, Mark W; Barber, Larry B

2013-01-15

The Potomac River basin is an area where a high prevalence of abnormalities such as testicular oocytes (TO), skin lesions, and mortality has been observed in smallmouth bass (SMB, Micropterus dolomieu). Previous research documented a variety of chemicals in regional streams, implicating chemical exposure as one plausible explanation for these biological effects. Six stream sites in the Potomac basin (and one out-of-basin reference site) were sampled to provide an assessment of chemicals in these streams. Potential early life-stage exposure to chemicals detected was assessed by collecting samples in and around SMB nesting areas. Target chemicals included those known to be associated with important agricultural and municipal wastewater sources in the Potomac basin. The prevalence and severity of TO in SMB were also measured to determine potential relations between chemistry and biological effects. A total of 39 chemicals were detected at least once in the discrete-water samples, with atrazine, caffeine, deethylatrazine, simazine, and iso-chlorotetracycline being most frequently detected. Of the most frequently detected chemicals, only caffeine was detected in water from the reference site. No biogenic hormones/sterols were detected in the discrete-water samples. In contrast, 100 chemicals (including six biogenic hormones/sterols) were found in a least one passive-water sample, with 25 being detected at all such samples. In addition, 46 chemicals (including seven biogenic hormones/sterols) were found in the bed-sediment samples, with caffeine, cholesterol, indole, para-cresol, and sitosterol detected in all such samples. The number of herbicides detected in discrete-water samples per site had a significant positive relation to TO(rank) (a nonparametric indicator of TO), with significant positive relations between TO(rank) and atrazine concentrations in discrete-water samples and to total hormone/sterol concentration in bed-sediment samples. Such significant correlations do not necessarily imply causation, as these chemical compositions and concentrations likely do not adequately reflect total SMB exposure history, particularly during critical life stages. Published by Elsevier B.V.

A comparison of QuantStudio™ 3D Digital PCR and ARMS-PCR for measuring plasma EGFR T790M mutations of NSCLC patients.

PubMed

Feng, Qin; Gai, Fei; Sang, Yaxiong; Zhang, Jie; Wang, Ping; Wang, Yue; Liu, Bing; Lin, Dongmei; Yu, Yang; Fang, Jian

2018-01-01

The AURA3 clinical trial has shown that advanced non-small cell lung cancer (NSCLC) patients with EGFR T790M mutations in circulating tumor DNA (ctDNA) could benefit from osimertinib. The aim of this study was to assess the usefulness of QuantStudio™ 3D Digital PCR System platform for the detection of plasma EGFR T790M mutations in NSCLC patients, and compare the performances of 3D Digital PCR and ARMS-PCR. A total of 119 Chinese patients were enrolled in this study. Mutant allele frequency of plasma EGFR T790M was detected by 3D Digital PCR, then 25 selected samples were verified by ARMS-PCR and four of them were verified by next generation sequencing (NGS). In total, 52.94% (69/119) had EGFR T790M mutations detected by 3D Digital PCR. In 69 positive samples, the median mutant allele frequency (AF) was 1.09% and three cases presented low concentration (AF <0.1%). Limited by the amount of plasma DNA, 17 samples (AF <2.5%) and eight samples (T790M-) were selected for verification by ARMS-PCR. Four of those samples were verified by NGS as a third verification method. Among the selected 17 positive cases, ten samples presented mutant allele frequency <0.5%, and seven samples presented intermediate mutant allele frequency (0.5% AF 2.5%). However, only three samples (3/17) were identified as positive by ARMS-PCR, namely, P6 (AF =1.09%), P7 (AF =2.09%), and P8 (AF =2.21%). It is worth mentioning that sample P9 (AF =2.05%, analyzed by 3D Digital PCR) was identified as T790M- by ARMS-PCR. Four samples were identified as T790M+ by both NGS and 3D Digital PCR, and typically three samples (3/4) presented at a low ratio (AF <0.5%). Our study demonstrated that 3D Digital PCR is a novel method with high sensitivity and specificity to detect EGFR T790M mutation in plasma.

A comparison of QuantStudio™ 3D Digital PCR and ARMS-PCR for measuring plasma EGFR T790M mutations of NSCLC patients

PubMed Central

Sang, Yaxiong; Zhang, Jie; Wang, Ping; Wang, Yue; Liu, Bing; Lin, Dongmei; Yu, Yang; Fang, Jian

2018-01-01

Background The AURA3 clinical trial has shown that advanced non-small cell lung cancer (NSCLC) patients with EGFR T790M mutations in circulating tumor DNA (ctDNA) could benefit from osimertinib. Purpose The aim of this study was to assess the usefulness of QuantStudio™ 3D Digital PCR System platform for the detection of plasma EGFR T790M mutations in NSCLC patients, and compare the performances of 3D Digital PCR and ARMS-PCR. Patients and methods A total of 119 Chinese patients were enrolled in this study. Mutant allele frequency of plasma EGFR T790M was detected by 3D Digital PCR, then 25 selected samples were verified by ARMS-PCR and four of them were verified by next generation sequencing (NGS). Results In total, 52.94% (69/119) had EGFR T790M mutations detected by 3D Digital PCR. In 69 positive samples, the median mutant allele frequency (AF) was 1.09% and three cases presented low concentration (AF <0.1%). Limited by the amount of plasma DNA, 17 samples (AF <2.5%) and eight samples (T790M-) were selected for verification by ARMS-PCR. Four of those samples were verified by NGS as a third verification method. Among the selected 17 positive cases, ten samples presented mutant allele frequency <0.5%, and seven samples presented intermediate mutant allele frequency (0.5% AF 2.5%). However, only three samples (3/17) were identified as positive by ARMS-PCR, namely, P6 (AF =1.09%), P7 (AF =2.09%), and P8 (AF =2.21%). It is worth mentioning that sample P9 (AF =2.05%, analyzed by 3D Digital PCR) was identified as T790M- by ARMS-PCR. Four samples were identified as T790M+ by both NGS and 3D Digital PCR, and typically three samples (3/4) presented at a low ratio (AF <0.5%). Conclusion Our study demonstrated that 3D Digital PCR is a novel method with high sensitivity and specificity to detect EGFR T790M mutation in plasma. PMID:29403309

Performance Indicators of the Top Basketball Players: Relations with Several Variables.

PubMed

Sindik, Josko

2015-09-01

The aim of this study was to determine the differences in performance indicators for top senior male basketball players, with respect to several independent variables: position in the team, total situation-related efficiency, age, playing experience and the time spent on the court within the game and during championship season. The final sample of participants was selected from all teams in A-1 Croatian men's basketball league. Significant differences have been found according to the players': position in the team, total situation-related efficiency, and in interactions of the position in the team / total situation-related efficiency and minutes spent on the court in a game / playing experience. The differences in the situation-related efficiency between players have not been found according to the players' age and the number of games played. Further research can be directed towards deeper analysis of the influence of more complex differentiated variables playing experience and time spent on the court in a game on situation-related efficiency in basketball.

Same-Day Identification and Antimicrobial Susceptibility Testing of Bacteria in Positive Blood Culture Broths Using Short-Term Incubation on Solid Medium with the MicroFlex LT, Vitek-MS, and Vitek2 Systems

PubMed Central

Ha, Jihye; Han, Geum Hee; Kim, Myungsook; Lee, Kyungwon

2018-01-01

Background Early and appropriate antibiotic treatment improves the clinical outcome of patients with septicemia; therefore, reducing the turn-around time for identification (ID) and antimicrobial susceptibility test (AST) results is essential. We established a method for rapid ID and AST using short-term incubation of positive blood culture broth samples on solid media, and evaluated its performance relative to that of the conventional method using two rapid ID systems and a rapid AST method. Methods A total of 254 mono-microbial samples were included. Positive blood culture samples were incubated on blood agar plates for six hours and identified by the MicroFlex LT (Bruker Daltonics) and Vitek-MS (bioMeriéux) systems, followed by AST using the Vitek2 System (bioMeriéux). Results The correct species-level ID rates were 82.3% (209/254) and 78.3% (199/254) for the MicroFlex LT and Vitek-MS platforms, respectively. For the 1,174 microorganism/antimicrobial agent combinations tested, the rapid AST method showed total concordance of 97.8% (1,148/1,174) with the conventional method, with a very major error rate of 0.5%, major error rate of 0.7%, and minor error rate of 1.0%. Conclusions Routine implementation of this short-term incubation method could provide ID results on the day of blood culture-positivity detection and one day earlier than the conventional AST method. This simple method will be very useful for rapid ID and AST of bacteria from positive blood culture bottles in routine clinical practice. PMID:29401558

Same-Day Identification and Antimicrobial Susceptibility Testing of Bacteria in Positive Blood Culture Broths Using Short-Term Incubation on Solid Medium with the MicroFlex LT, Vitek-MS, and Vitek2 Systems.

PubMed

Ha, Jihye; Hong, Sung Kuk; Han, Geum Hee; Kim, Myungsook; Yong, Dongeun; Lee, Kyungwon

2018-05-01

Early and appropriate antibiotic treatment improves the clinical outcome of patients with septicemia; therefore, reducing the turn-around time for identification (ID) and antimicrobial susceptibility test (AST) results is essential. We established a method for rapid ID and AST using short-term incubation of positive blood culture broth samples on solid media, and evaluated its performance relative to that of the conventional method using two rapid ID systems and a rapid AST method. A total of 254 mono-microbial samples were included. Positive blood culture samples were incubated on blood agar plates for six hours and identified by the MicroFlex LT (Bruker Daltonics) and Vitek-MS (bioMeriéux) systems, followed by AST using the Vitek2 System (bioMeriéux). The correct species-level ID rates were 82.3% (209/254) and 78.3% (199/254) for the MicroFlex LT and Vitek-MS platforms, respectively. For the 1,174 microorganism/antimicrobial agent combinations tested, the rapid AST method showed total concordance of 97.8% (1,148/1,174) with the conventional method, with a very major error rate of 0.5%, major error rate of 0.7%, and minor error rate of 1.0%. Routine implementation of this short-term incubation method could provide ID results on the day of blood culture-positivity detection and one day earlier than the conventional AST method. This simple method will be very useful for rapid ID and AST of bacteria from positive blood culture bottles in routine clinical practice. © The Korean Society for Laboratory Medicine

Legionella spp. in dental unit waterlines.

PubMed

Sedlata Juraskova, E; Sedlackova, H; Janska, J; Holy, O; Lalova, I; Matouskova, I

2017-01-01

To determine the current presence of Legionella spp. in the output water of dental unit waterlines (DUWLs) and examine its mitigation by disinfection at the Institute of Dentistry and Oral Sciences, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc. The first stage of our survey involved collecting samples of DUWL output water from 50 dental chair units (DCUs), and 2 samples of the incoming potable water. In October 2015, a one-time disinfection (1 % Stabimed) of DUWLs was conducted. This was followed by collecting 10 control samples (survey stage 2). From the total of 50 samples (survey stage 1), 18 samples (36.0 %) tested positive for Legionella spp. Following the disinfection, nine of the ten samples no longer showed any presence of Legionella. Based on culture results, the one-time disinfection (1 % Stabimed) was effective. We are unable to comment on the duration of positive effect of disinfection on the occurrence of Legionella spp. in the outlet water. It was a one-time survey (Tab. 2, Ref. 32).

First isolation and molecular characterization of Ehrlichia canis in Costa Rica, Central America.

PubMed

Romero, L E; Meneses, A I; Salazar, L; Jiménez, M; Romero, J J; Aguiar, D M; Labruna, M B; Dolz, G

2011-08-01

The present study investigated Ehrlichia species in blood samples from dogs suspected of clinical ehrlichiosis, using molecular and isolation techniques in cell culture. From a total of 310 canine blood samples analyzed by 16S rRNA nested PCR, 148 (47.7%) were positive for Ehrlichia canis. DNA from Ehrlichia chaffeensis or Ehrlichia ewingii was not detected in any sample using species-specific primers in separated reactions. Leukocytes from five PCR-positive dogs were inoculated into DH82 cells; successful isolation of E. canis was obtained in four samples. Partial sequence of the dsb gene of eight canine blood samples (including the five samples for in vitro isolation) was obtained by PCR and their analyses through BLAST showed 100% of identity with the corresponding sequence of E. canis in GenBank. This study represents the first molecular diagnosis, isolation, and molecular characterization of E. canis in dogs from Costa Rica. Copyright © 2010 Elsevier Ltd. All rights reserved.

Comparative performance of aldolase and lactate dehydrogenase rapid diagnostic tests in Plasmodium vivax detection

PubMed Central

2014-01-01

Background Misdiagnosis of malaria by commercial rapid diagnostic tests (RDTs) is a major cause of concern in the diagnosis of malaria. This retrospective study was aimed at assessing the relative performance of four RDTs with emphasis on the detection of two Plasmodium vivax antigens: aldolase and lactate dehydrogenase (LDH). Methods Three commercially available Plasmodium LDH or aldolase antigen detection kits (One Step Malaria P.f/P.v, ParaHit Total ver. 1.0, SD Bioline Malaria) and an anti-P. vivax aldolase-specific monoclonal antibody (mAb) pair 1C3-12 F10 were evaluated with P. vivax positive as well as non-P. vivax samples and healthy samples using blood smear examination as standard. Each test was read according to the manufacturer’s instructions. Results MAb 1C3-12 F10 pair targeting P. vivax-specific aldolase exhibited very good specificity and sensitivity of 100 and 97.4%, respectively. Positive predictive value (PPV) and negative predictive value (NPV) of 100 and 99.5%, respectively, were also observed. The anti-P. vivax LDH in the One-Step Malaria P.f/P.v test showed sensitivity, specificity, PPV and NPV of 93.5, 98.0, 88.9 and 98.8%, respectively. ParaHit Total ver. 1.0 targeting the pan-aldolase antigen showed sensitivity, specificity of 97.4 and 99.6%, respectively. PPV and NPV were both 99.5%. SD Bioline had sensitivity, specificity, PPV and NPV of 93.5, 100, 100 and 98.8%, respectively. The overall sensitivity and specificity of all four RDTs were acceptable, especially for the aldolase detection tests. Five (6.5%) of the P. vivax-positive samples (n = 77) that were confirmed by microscopic examination as well as the two aldolase detection RDTs (mAb 1C3-12 F10 and ParaHit Total ver.1.0) were undetected by the two LDH detection RDTs (One Step Malaria P.f/P.v and SD Bioline). Similarly, two positive samples (2.6%) that were positively confirmed by the LDH detection RDTs were also undetected by the aldolase detection test kits. Conclusion Aldolase and LDH antigens perform differently in different P. vivax samples; hence there is a high risk of misdiagnosis when monoclonal antibodies are used against only one particular antigen in the test. A combination of both aldolase and LDH in RDTs for the rapid diagnosis of P. vivax will enhance the sensitivity of the assay and reduce misdiagnosis. PMID:25015737

[Seropositivity of Borrelia burgdorferi in risky groups in Van region, Turkey].

PubMed

Parlak, Mehmet; Bayram, Yasemin; Çıkman, Aytekin; Ceylan, Nesrin; Berktaş, Mustafa

2015-07-01

Lyme borreliosis, which is more prevalent in the northern hemisphere, is the most common tick-borne contagious disease among people living in the North America and Europe. The causative agent of Lyme borreliosis, Borrelia burgdorferi, is transmitted by the bites of ticks of the genus Ixodes. In Turkey, the seroprevalence of Lyme disease is increased in regions where ticks and tick-bite cases are prevalent. The present study aimed to determine the seroprevalence of Lyme borreliosis in people at risk, living in the rural areas of Van province, which is located in the eastern region of Turkey. No previous study on this topic has been performed in our province. The study included a total of 446 subjects (mean age: 39.6±15.5 years), of them 139 were male and 307 were female, living in the rural areas of Van province between January 2012 and July 2012. The serum samples collected from participants after informed consent were screened for the presence of B.burgdorferi IgG antibodies by ELISA method. Western blot (WB) method was used for the confirmation of positive or borderline positive samples, and also for the investigation of IgM antibodies. During the study, the individuals from whom samples were taken, were questioned whether they have ever been exposed to tick or insect bite. B.burgdorferi IgG positivity was detected in 17 (3.8%) of the cases, whereas it was within the limit values in 14 cases. A total of 31 samples which yielded positive and borderline positive results were retested by WB and 4 (12.9%) were detected as positive while 10 (32.3%) of the samples were indeterminate. B.burgdorferi IgM antibody positivity was not detected in any of the samples. Considering the WB as reference method, the rate of B.burgdorferi IgG seropositivity was estimated as 0.9% (4/446). Three of these four cases were defined as tick or insect bites. The seroprevalence rate of B.burgdorferi detected in the present study was low as compared to the results of the other studies reported from Turkey. The reason of this result might be from the geographical characteristics and the differences of tick fauna in our region. As a result, it was concluded that our province is not endemic for Lyme borreliosis, however for the reduction of tick exposure, emphasis must be placed on preventive health services for the individuals at risk.

Virus occupational exposure in solid waste processing facilities.

PubMed

Carducci, Annalaura; Federigi, Ileana; Verani, Marco

2013-11-01

It is well known that workers involved in the management of solid waste are at risk of exposure to bioaerosol, which is generally studied in relation to bacteria, fungi, and endotoxins. However, to date, there have been no reports on the incidence of work-related infectious diseases. To determine if occupational exposure to viruses occurs upon exposure to waste-related activities, monitoring was carried out in a landfill, a waste recycling plant, an incineration plant, and a waste collection vehicles. Air and surfaces were sampled and analyzed for torque teno virus (TTV), human adenovirus (HAdV), norovirus, rotavirus, and enterovirus using polymerase chain reaction (PCR)-based techniques. Positivity was confirmed by sequencing and quantification with real-time PCR; infectivity was also tested for culturable viruses. Samples were analyzed in parallel for mean total bacterial and fungi counts in both the summer and winter. In total, 30% (12/40) of air and 13.5% (5/37) of surface samples collected in plants were positive for HAdV and TTV. Among the eight HAdV-positive samples, six (75%), revealed in landfill and recycling plant air and in incinerator and waste vehicles surfaces, were able to replicate in cell culture and were subsequently confirmed as infective. The frequency of detection of virus-positive samples was similar in both seasons, but with evident differences in the type of virus detected: TTV and HAdV were more frequently detected in the summer and winter, respectively. The area of highest viral contamination was the paper selection landfill. Fungi and bacterial contamination did not correlate with viral presence or concentration. In conclusion, we evidence that working with solid and liquid waste can lead to infectious viruses, included in Group 2 of the European Directive 90/679/CEE pathogens list; thus, further investigation on the sources and routes of contamination is needed in order to assess the occupational risk.

Effect of different postoperative limb positions on blood loss and range of motion in total knee arthroplasty: An updated meta-analysis of randomized controlled trials.

PubMed

Wu, Yuangang; Yang, Timin; Zeng, Yi; Si, Haibo; Li, Canfeng; Shen, Bin

2017-01-01

Postoperative limb positioning has been reported to be an efficient and simple way to reduce blood loss and improve range of motion following total knee arthroplasty (TKA). This meta-analysis was designed to compare the effectiveness of two different limb positions in primary TKA. A meta-analysis of the PubMed, CENTRAL, Web of Science, EMBASE and Google Search Engine electronic databases was performed. In this meta-analysis, two postoperative limb positions were considered: mild-flexion (flexion less than 60°) and high-flexion (flexion at 60° or more). The subgroups were analysed using RevMan 5.3. Nine RCTs were included with a total sample size of 913 patients. The mild- and high-flexion positions significantly reduced postoperative total blood loss (P = 0.04 and P = 0.01; respectively). Subgroup analysis indicated that knee flexion significantly reduced hidden blood loss when the knee was fixed in mild-flexion (P = 0.0004) and significantly reduced transfusion requirements (P = 0.03) and improved range of motion (ROM) (P < 0.00001) when the knee was fixed in high-flexion. However, the rates of wound-related infection, deep venous thrombosis (DVT) and pulmonary embolism (PE) did not significantly differ between the two flexion groups. This meta-analysis suggests that mild- and high-flexion positions have similar efficacy in reducing total blood loss. In addition, subgroup analysis indicates that the mild-flexion position is superior in decreasing hidden blood loss compared with high-flexion; the high-flexion position is superior to mild-flexion in reducing transfusion requirements and improving postoperative ROM. Thus, the use of the high-flexion position is a viable option to reduce blood loss in patients following primary TKA without increasing the risk of wound-related infection, DVT or PE. Copyright © 2016 IJS Publishing Group Ltd. Published by Elsevier Ltd. All rights reserved.

The metabolic power and energetic demands of elite Gaelic football match play.

PubMed

Malone, Shane; Solan, Barry; Collins, Kieran; Doran, Dominic

2017-05-01

Metabolic power has not yet been investigated within elite Gaelic football. The aim of the current investigation was to compare the metabolic power demands between positional groups and examine the temporal profile of elite Gaelic football match play. Global positional satellite system (GPS) data were collected from 50 elite Gaelic football players from 4 inter-county teams during 35 elite competitive matches over a three season period. A total of 351 complete match samples were obtained for final analysis. Players were categorized based on positional groups; full-back, half-back, midfield, half-forward and full-forward. Instantaneous raw velocity data was obtained from the GPS and exported to a customized spreadsheet which provided estimations of both speed based, derived metabolic power and energy expenditure variables (total distance, high speed distance, average metabolic power, high power distance and total energy expenditure). Match mean distance was 9222±1588 m, reflective of an average metabolic power of 9.5-12.5 W·kg-1, with an average energy expenditure of 58-70 Kj·kg-1 depending on position. There were significant differences between positional groups for both speed-based and metabolic power indices. Midfielders covered more total and high-speed distance, as well as greater average and overall energy expenditure compared to other positions (P<0.001). A reduction in total, high-speed, and high-power distance, as well as average metabolic power throughout the match (P<0.001) was observed. Positional differences exist for both metabolic power and traditional running based variables. The middle three positions (midfield, half-back and half-forward) possess greater activity profiles when compared to other positional groups. The reduction in metabolic power and traditional running based variables are comparable across match play. The current study demonstrates that metabolic power may contribute to our understanding of Gaelic football match-play.

Marine Bacteria Cause False-Positive Results in the Colilert-18 Rapid Identification Test for Escherichia coli in Florida Waters

PubMed Central

Pisciotta, John M.; Rath, Damon F.; Stanek, Paul A.; Flanery, D. Michael; Harwood, Valerie J.

2002-01-01

The Colilert-18 system for enumeration of total coliforms and Escherichia coli is approved by the U.S. Environmental Protection Agency for use in drinking water analysis and is also used by various agencies and research studies for enumeration of indicator organisms in fresh and saline waters. During monitoring of Pinellas County, Fla., marine waters, estimates of E. coli numbers (by Colilert-18) frequently exceeded fecal coliform counts (by membrane filtration) by 1 to 3 orders of magnitude. Samples from freshwater sites did not display similar discrepancies. Fecal coliforms, including E. coli, could be cultured from 100% of yellow fluorescent wells (denoting E. coli-positive results) inoculated with freshwater samples but could be cultured from only 17.1% of the “positive” wells inoculated with marine samples. Ortho-nitrophenyl-β-d-galactopyranoside (ONPG)-positive or 4-methylumbelliferyl-β-d-glucuronide (MUG)-positive noncoliform bacteria were readily cultured from Colilert-18 test wells inoculated with marine samples. Filtered cell-free seawater did not cause false positives. Coculture preparations of as few as 5 CFU of Vibrio cholerae (ONPG positive) and Providencia sp. (MUG positive) ml−1 inoculated into Colilert-18 caused false-positive E. coli results. Salinity conditions influenced coculture results, as the concentration of coculture inoculum required to cause false positives in most wells increased from about 5 CFU ml−1 in seawater diluted 1:10 with freshwater to ≈5,000 CFU ml−1 in seawater diluted 1:20 with freshwater. Estimated E. coli numbers in various marine water samples processed at the 1:10 dilution ranged from 10 to 7,270 CFU·100 ml−1, while E. coli numbers in the same samples processed at the 1:20 dilution did not exceed 40 CFU·100 ml−1. The lower estimates of E. coli numbers corresponded well with fecal coliform counts by membrane filtration. This study indicates that assessment of E. coli in subtropical marine waters by Colilert-18 is not accurate when the recommended 1:10 sample dilution is used. The results suggest that greater dilution may diminish the false-positive problem, but further study of this possibility is recommended. PMID:11823188

Stellar Stream Candidates in the Solar Neighborhood Found in the LAMOST DR3 and TGAS

NASA Astrophysics Data System (ADS)

Liang, X. L.; Zhao, J. K.; Oswalt, T. D.; Chen, Y. Q.; Zhang, L.; Zhao, G.

2017-08-01

We have cross-matched the LAMOST DR3 with the Gaia DR1 TGAS catalogs and obtained a sample of 166,827 stars with reliable kinematics. A technique based on the wavelet transform was applied to detect significant overdensities in velocity space among five subsamples divided by spatial position. In total, 16 significant overdensities of stars with very similar kinematics were identified. Among these, four are new stream candidates and the rest are previously known groups. Both the U-V velocity and metallicity distributions of the local sample show a clear gap between the Hercules structure and the Hyades-Pleiades structure. The U-V positions of these peaks shift with the spatial position. Following a description of our analysis, we speculate on possible origins of our stream candidates.

Dose-Response Analysis of RNA-Seq Profiles in Archival ...

EPA Pesticide Factsheets

Use of archival resources has been limited to date by inconsistent methods for genomic profiling of degraded RNA from formalin-fixed paraffin-embedded (FFPE) samples. RNA-sequencing offers a promising way to address this problem. Here we evaluated transcriptomic dose responses using RNA-sequencing in paired FFPE and frozen (FROZ) samples from two archival studies in mice, one 20 years old. Experimental treatments included 3 different doses of di(2-ethylhexyl)phthalate or dichloroacetic acid for the recently archived and older studies, respectively. Total RNA was ribo-depleted and sequenced using the Illumina HiSeq platform. In the recently archived study, FFPE samples had 35% lower total counts compared to FROZ samples but high concordance in fold-change values of differentially expressed genes (DEGs) (r2 = 0.99), highly enriched pathways (90% overlap with FROZ), and benchmark dose estimates for preselected target genes (2% difference vs FROZ). In contrast, older FFPE samples had markedly lower total counts (3% of FROZ) and poor concordance in global DEGs and pathways. However, counts from FFPE and FROZ samples still positively correlated (r2 = 0.84 across all transcripts) and showed comparable dose responses for more highly expressed target genes. These findings highlight potential applications and issues in using RNA-sequencing data from FFPE samples. Recently archived FFPE samples were highly similar to FROZ samples in sequencing q

Status of microbial diversity in agroforestry systems in Tamil Nadu, India.

PubMed

Radhakrishnan, Srinivasan; Varadharajan, Mohan

2016-06-01

Soil is a complex and dynamic biological system. Agroforestry systems are considered to be an alternative land use option to help and prevent soil degradation, improve soil fertility, microbial diversity, and organic matter status. An increasing interest has emerged with respect to the importance of microbial diversity in soil habitats. The present study deals with the status of microbial diversity in agroforestry systems in Tamil Nadu. Eight soil samples were collected from different fields in agroforestry systems in Cuddalore, Villupuram, Tiruvanamalai, and Erode districts, Tamil Nadu. The number of microorganisms and physico-chemical parameters of soils were quantified. Among different microbial population, the bacterial population was recorded maximum (64%), followed by actinomycetes (23%) and fungi (13%) in different samples screened. It is interesting to note that the microbial population was positively correlated with the physico-chemical properties of different soil samples screened. Total bacterial count had positive correlation with soil organic carbon (C), moisture content, pH, nitrogen (N), and micronutrients such as Iron (Fe), copper (Cu), and zinc (Zn). Similarly, the total actinomycete count also showed positive correlations with bulk density, moisture content, pH, C, N, phosphorus (P), potassium (K), calcium (Ca), copper (Cu), magnesium (Mg), manganese (Mn), and zinc (Zn). It was also noticed that the soil organic matter, vegetation, and soil nutrients altered the microbial community under agroforestry systems. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Escherichia coli O104 in Feedlot Cattle Feces: Prevalence, Isolation and Characterization.

PubMed

Shridhar, Pragathi B; Noll, Lance W; Shi, Xiaorong; Cernicchiaro, Natalia; Renter, David G; Bai, J; Nagaraja, T G

2016-01-01

Escherichia coli O104:H4, an hybrid pathotype of Shiga toxigenic and enteroaggregative E. coli, involved in a major foodborne outbreak in Germany in 2011, has not been detected in cattle feces. Serogroup O104 with H type other than H4 has been reported to cause human illnesses, but their prevalence and characteristics in cattle have not been reported. Our objectives were to determine the prevalence of E. coli O104 in feces of feedlot cattle, by culture and PCR detection methods, and characterize the isolated strains. Rectal fecal samples from a total of 757 cattle originating from 29 feedlots were collected at a Midwest commercial slaughter plant. Fecal samples, enriched in E. coli broth, were subjected to culture and PCR methods of detection. The culture method involved immunomagnetic separation with O104-specific beads and plating on a selective chromogenic medium, followed by serogroup confirmation of pooled colonies by PCR. If pooled colonies were positive for the wzxO104 gene, then colonies were tested individually to identify wzxO104-positive serogroup and associated genes of the hybrid strains. Extracted DNA from feces were also tested by a multiplex PCR to detect wzxO104-positive serogroup and associated major genes of the O104 hybrid pathotype. Because wzxO104 has been shown to be present in E. coli O8/O9/O9a, wzxO104-positive isolates and extracted DNA from fecal samples were also tested by a PCR targeting wbdDO8/O9/O9a, a gene specific for E. coli O8/O9/O9a serogroups. Model-adjusted prevalence estimates of E. coli O104 (positive for wzxO104 and negative for wbdDO8/O9/O9a) at the feedlot level were 5.7% and 21.2%, and at the sample level were 0.5% and 25.9% by culture and PCR, respectively. The McNemar's test indicated that there was a significant difference (P < 0.01) between the proportions of samples that tested positive for wzxO104 and samples that were positive for wzxO104, but negative for wbdDO8/O9/O9a by PCR and culture methods. A total of 143 isolates, positive for the wzxO104, were obtained in pure culture from 146 positive fecal samples. Ninety-two of the 143 isolates (64.3%) also tested positive for the wbdDO8/O9/O9a, indicating that only 51 (35.7%) isolates truly belonged to the O104 serogroup (positive for wzxO104 and negative for wbdDO8/O9/O9a). All 51 isolates tested negative for eae, and 16 tested positive for stx1 gene of the subtype 1c. Thirteen of the 16 stx1-positive O104 isolates were from one feedlot. The predominant serotype was O104:H7. Pulsed-field gel electrophoresis analysis indicated that stx1-positive O104:H7 isolates had 62.4% homology to the German outbreak strain and 67.9% to 77.5% homology to human diarrheagenic O104:H7 strains. The 13 isolates obtained from the same feedlot were of the same PFGE subtype with 100% Dice similarity. Although cattle do not harbor the O104:H4 pathotype, they do harbor and shed Shiga toxigenic O104 in the feces and the predominant serotype was O104:H7.

Airborne Bacteria in an Urban Environment

PubMed Central

Mancinelli, Rocco L.; Shulls, Wells A.

1978-01-01

Samples were taken at random intervals over a 2-year period from urban air and tested for viable bacteria. The number of bacteria in each sample was determined, and each organism isolated was identified by its morphological and biochemical characteristics. The number of bacteria found ranged from 0.013 to 1.88 organisms per liter of air sampled. Representatives of 19 different genera were found in 21 samples. The most frequently isolated organisms and their percent of occurence were Micrococcus (41%), Staphylococcus (11%), and Aerococcus (8%). The bacteria isolated were correlated with various weather and air pollution parameters using the Pearson product-moment correlation coefficient method. Statistically significant correlations were found between the number of viable bacteria isolated and the concentrations of nitric oxide (−0.45), nitrogen dioxide (+0.43), and suspended particulate pollutants (+0.56). Calculated individually, the total number of Micrococcus, Aerococcus, and Staphylococcus, number of rods, and number of cocci isolated showed negative correlations with nitric oxide and positive correlations with nitrogen dioxide and particulates. Statistically significant positive correlations were found between the total number of rods isolated and the concentration of nitrogen dioxide (+0.54) and the percent relative humidity (+0.43). The other parameters tested, sulfur dioxide, hydrocarbons, and temperature, showed no significant correlations. Images PMID:677875

Molecular Phylogenetic Diversity and Spatial Distribution of Bacterial Communities in Cooling Stage during Swine Manure Composting

PubMed Central

Guo, Yan; Zhang, Jinliang; Yan, Yongfeng; Wu, Jian; Zhu, Nengwu; Deng, Changyan

2015-01-01

Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and subsequent sub-cloning and sequencing were used in this study to analyze the molecular phylogenetic diversity and spatial distribution of bacterial communities in different spatial locations during the cooling stage of composted swine manure. Total microbial DNA was extracted, and bacterial near full-length 16S rRNA genes were subsequently amplified, cloned, RFLP-screened, and sequenced. A total of 420 positive clones were classified by RFLP and near-full-length 16S rDNA sequences. Approximately 48 operational taxonomic units (OTUs) were found among 139 positive clones from the superstratum sample; 26 among 149 were from the middle-level sample and 35 among 132 were from the substrate sample. Thermobifida fusca was common in the superstratum layer of the pile. Some Bacillus spp. were remarkable in the middle-level layer, and Clostridium sp. was dominant in the substrate layer. Among 109 OTUs, 99 displayed homology with those in the GenBank database. Ten OTUs were not closely related to any known species. The superstratum sample had the highest microbial diversity, and different and distinct bacterial communities were detected in the three different layers. This study demonstrated the spatial characteristics of the microbial community distribution in the cooling stage of swine manure compost. PMID:25925066

Prevalence and Characterization of Salmonella in Animal Meals Collected from Rendering Operations.

PubMed

Jiang, Xiuping

2016-06-01

As part of the Salmonella Education Reduction Program, the Animal Protein Producers Industry initiated a yearlong microbiological survey of animal meals from 1 January to 31 December 2010. The types of animal meals included poultry meal, pork and beef crax, meat meal, meat and bone meal, feather meal, blood meal, and fish meal from a variety of rendering operations (n = 65). Salmonella was positive in 731 (8.3%) of 8,783 analyzed samples, with contamination rates as 1.0, 33.2, and 21.3% from samples collected right after press, being loaded out, or unidentified, respectively. The randomly selected positive Salmonella samples (n = 100) representing 1.1% of the total samples tested were enumerated by the most-probable-number (MPN) method. The Salmonella contamination level ranged from <0.03 (below the detection limit) to 240 MPN/g with a median MPN per gram of 0.036. Among 102 Salmonella isolates from those 100 positive samples, a total of 42 Salmonella serotypes or groups were identified with Montevideo (13%), Senftenberg (11%), Mbandaka (7%), Orion (7%), Livingstone (6%), Tennessee (4%), Infantis (4%), Cerro (4%), and group C1 (4%) as the most predominant ones. Those Salmonella isolates were further analyzed for antimicrobial resistance to the 15 most common antibiotics by using the National Antimicrobial Resistance Monitoring System gram-negative plate. Most Salmonella isolates (n = 94) were sensitive to all antibiotics tested, with seven isolates resistant to one antibiotic and one resistant to seven antibiotics. Clearly, the prevalence of Salmonella in animal meals declined compared with previous surveys, and none of the Salmonella serotypes concerning target animal health were isolated. In addition, most Salmonella isolates remained susceptible to the majority of the 15 most commonly used antibiotics.

Prevalence and genetic characterization of Toxoplasma gondii in donkeys in northeastern China.

PubMed

Zhang, Xiao-Xuan; Shi, Wei; Zhang, Nian-Zhang; Shi, Kun; Li, Jian-Ming; Xu, Peng; Zhao, Quan; Du, Rui

2017-10-01

Toxolasma gondii is one of the most important obligate intracellular protozoan parasites. Recently, toxoplasmosis is of increasing concerns because T. gondii not only has a worldwide distribution but also can infect virtually all warm-blooded hosts including donkeys. However, limited information is available concerning the genetic characterization of T. gondii in donkeys in northeastern China. In this study, a total of 302 brain tissue samples collected from donkeys from Jilin (n=108) and Liaoning (n=194) provinces, northeastern China, were examined for T. gondii infection by semi-nested PCR of B1 gene. The positive samples were genotyped at 11 loci (i.e., SAG1, alternative SAG2, 5'-and 3'-SAG2, SAG3, L358, BTUB, c22-8, GRA6, c29-2, PK1 and Apico) using polymerase chain reaction-restriction fragment length polymorphism technology. Of 302 brain tissue samples, 19 (6.29%) were PCR-positive for T. gondii. The prevalence rates of T. gondii were 6.48% (7/108) and 6.19% (12/194) in Jilin Province and Liaoning Province, respectively. The prevalence of T. gondii in different season groups varied from 5.56% to 7.41%. The prevalence of T. gondii in "Cage-free" donkeys was higher than that in "Captive" donkeys. Of the 19 positive samples, only two isolates were successfully genotyped at all loci, three were genotyped at 9 loci. In total, four samples belong to ToxoDB genotype #9, one belong to ToxoDB genotype #10. This is firstly characterized the T. gondii isolates from donkeys in northeastern China. The results of the present study will improve the information of the distribution of T. gondii genotypes in China. Copyright © 2017. Published by Elsevier B.V.

Histopathological and immunohistochemical approaches for the diagnosis of Pasteurellosis in swine population of Punjab.

PubMed

Bhat, Payal; Singh, Nittin Dev; Leishangthem, Geeta Devi; Kaur, Amninder; Mahajan, Vishal; Banga, Harmanjit Singh; Brar, Rajinder Singh

2016-09-01

Infectious porcine bronchopneumonia, caused by Pasteurella multocida , is a widespread disease of major economic significance. Thus, the aim of the present study was to diagnose swine Pasteurellosis using gross, histopathological, and immunopathological approaches in the swine population of Punjab and to compare the efficacy of immunohistochemical (IHC) techniques with conventional diagnostic techniques. A total of 71 adult swine lung samples showing gross pneumonic changes were collected along with the associated lymph nodes to carry out the study. The collected samples were then processed for histopathological and IHC studies. Out of the total 71 lung samples, 26 samples were found to be suspected for Pasteurellosis as per the microscopic changes observed, and out of these 26 samples, 16 cases were confirmed to be positive for Pasteurellosis by IHC. Varied macroscopic changes noted in lungs were pneumonic patches with consolidation of many lobes, congestion, and focal hemorrhages. Main lesions associated with lymph nodes were its enlargement and hemorrhages. Histologically, the lung showed fibrinous and suppurative bronchopneumonia, multifocal suppuration, thickening of septa with fibrin combined with cellular infiltration and edema. The higher IHC expression of P. multocida was seen in the bronchial epithelium besides in alveolar and bronchial exudate. Moreover, on comparing the histopathological and IHC scores which were calculated on the basis of characteristic microscopic lesions and number of antigen positive cells, respectively, a significant positive correlation (r=0.4234) was found. It was concluded that swine population of Punjab is having P. multocida infection. The gross and histopathological lesions can be helpful in the preliminary diagnosis of Pasteurellosis but needs to be supplemented by other immunodiagnostic tests. Moreover, IHC technique proved to be a specific, reliable, precise, and rapid technique to supplement these conventional methods of diagnosis for Pasteurellosis.

Water quality and diversity of yeasts from tropical lakes and rivers from the Rio Doce basin in Southeastern Brazil

PubMed Central

Medeiros, Adriana O.; Missagia, Beatriz S.; Brandão, Luciana R.; Callisto, Marcos; Barbosa, Francisco A. R.; Rosa, Carlos A.

2012-01-01

Yeast communities were assessed in 14 rivers and four lakes from the Doce River basin in Brazil, during the rainy and dry seasons of the years 2000 and 2001. Water samples were collected at the subsurface in all sites. The following physical and chemical parameters were measured: temperature, dissolved oxygen, pH, electrical conductivity, total phosphorus, ortho-phosphate, ammonium, nitrate, nitrite and total nitrogen and the counts of faecal coliforms and heterotrophic bacteria were carried out to characterize the aquatic environmental sampled. The yeast counts were higher in aquatic environments with the highest counts of coliform and heterotrophic bacteria. These environments receive a high influx of domestic and industrial waste. A total of 317 isolates identified in forty eight yeast species were recorded in the sites sampled and the specie Aureobasidium pullulans were found in eleven out of eighteen sites sampled and some opportunistic pathogens such as the yeast species Candida krusei were isolated only in the polluted rivers with a positive correlation with the biotic and abiotic parameters that indicate sewage contamination. PMID:24031990

Population diversity of ammonium oxidizers investigated by specific PCR amplification

USGS Publications Warehouse

Ward, B.B.; Voytek, M.A.; Witzel, K.-P.

1997-01-01

The species composition of ammonia-oxidizing bacteria in aquatic environments was investigated using PCR primers for 16S rRNA genes to amplify specific subsets of the total ammonia-oxidizer population. The specificity of the amplification reactions was determined using total genomic DNA from known nitrifying strains and non-nitrifying strains identified as having similar rDNA sequences. Specificity of amplification was determined both for direct amplification, using the nitrifier specific primers, and with nested amplification, in which the nitrifier primers were used to reamplify a fragment obtained from direct amplification with Eubacterial universal primers. The present level of specificity allows the distinction between Nitrosomonas europaea, Nitrosomonas sp. (marine) and the other known ammonia-oxidizers in the beta subclass of the Proteobacteria. Using total DNA extracted from natural samples, we used direct amplification to determine presence/absence of different species groups. Species composition was found to differ among depths in vertical profiles of lake samples and among samples and enrichments from various other aquatic environments. Nested PCR yielded several more positive reactions, which implies that nitrifier DNA was present in most samples, but often at very low levels.

The relationship between personalities and self-report positive driving behavior in a Chinese sample

PubMed Central

Sun, Xianghong; Zhang, Kan

2018-01-01

Driving behaviors play an important role in accident involvement. Concretely speaking, aberrant driving behaviors would cause more accidents, and oppositely positive driving behaviors would promote to build safety traffic environment. The main goals of this study were to explore the positive driving behavior and its relationship with personality in a Chinese sample. A total of 421 licensed drivers (286 male and 135 female) from Beijing, China completed the Positive Driver Behavior Scale (PDBS), the Driver Behavior Questionnaire (DBQ), the Dula Dangerous Driving Index (DDDI) and the Big Five Inventory (BFI) on a voluntary and anonymous basis. The results showed that the Chinese version of the PDBS has both reliability and validity and that the PDBS was significantly correlated with the BFI. Specifically, the PDBS was negatively correlated with neuroticism (r = -0.38) and positively correlated with extraversion, agreeableness, conscientiousness and openness to experience (the correlation coefficient ranged from 0.36 to 0.55). In contrast with previous research, age was negatively correlated with the PDBS (r = -0.38) in our sample, which may have resulted from less driving experience or a lack of available cognitive resources. PMID:29324823

The relationship between personalities and self-report positive driving behavior in a Chinese sample.

PubMed

Shen, Biying; Qu, Weina; Ge, Yan; Sun, Xianghong; Zhang, Kan

2018-01-01

Driving behaviors play an important role in accident involvement. Concretely speaking, aberrant driving behaviors would cause more accidents, and oppositely positive driving behaviors would promote to build safety traffic environment. The main goals of this study were to explore the positive driving behavior and its relationship with personality in a Chinese sample. A total of 421 licensed drivers (286 male and 135 female) from Beijing, China completed the Positive Driver Behavior Scale (PDBS), the Driver Behavior Questionnaire (DBQ), the Dula Dangerous Driving Index (DDDI) and the Big Five Inventory (BFI) on a voluntary and anonymous basis. The results showed that the Chinese version of the PDBS has both reliability and validity and that the PDBS was significantly correlated with the BFI. Specifically, the PDBS was negatively correlated with neuroticism (r = -0.38) and positively correlated with extraversion, agreeableness, conscientiousness and openness to experience (the correlation coefficient ranged from 0.36 to 0.55). In contrast with previous research, age was negatively correlated with the PDBS (r = -0.38) in our sample, which may have resulted from less driving experience or a lack of available cognitive resources.

Detection of Human Immunodeficiency Virus Type 1 (HIV-1) RNA in Pools of Sera Negative for Antibodies to HIV-1 and HIV-2

PubMed Central

Morandi, Pierre-Alain; Schockmel, Gérard A.; Yerly, Sabine; Burgisser, Philippe; Erb, Peter; Matter, Lukas; Sitavanc, Radan; Perrin, Luc

1998-01-01

A total of 234 pools were prepared from 10,692 consecutive serum samples negative for antibodies to human immunodeficiency virus type 1 (HIV-1) and HIV-2 collected at five virological laboratories (average pool size, 45 serum samples). Pools were screened for the presence of HIV-1 RNA by a modified commercial assay (Amplicor HIV-1 Monitor test) which included an additional polyethylene glycol (PEG) precipitation step prior to purification of viral RNA (PEG Amplicor assay). The sensitivity of this assay for HIV-1 RNA detection in individual serum samples within pools matches that of standard commercial assays for individual serum samples, i.e., 500 HIV-1 RNA copies per ml. Five pools were identified as positive, and each one contained one antibody-negative, HIV-1 RNA-positive serum sample, corresponding to an average of 1 infected sample per 2,138 serum samples. Retrospective analysis revealed that the five HIV-1 RNA-positive specimens originated from individuals who had symptomatic primary HIV-1 infection at the time of sample collection and who were also positive for p24 antigenemia. We next assessed the possibility of performing the prepurification step by high-speed centrifugation (50,000 × g for 80 min) of 1.5-ml pools containing 25 μl of 60 individual serum samples, of which only 1 contained HIV-1 RNA (centrifugation Amplicor assay). The sensitivity of this assay also matches the sensitivities of standard commercial assays for HIV-1 RNA detection in individual serum samples. The results demonstrate that both assays with pooled sera can be applied to the screening of large numbers of serum samples in a time- and cost-efficient manner. PMID:9620372

Access to environmental resources and physical activity levels of adults in Hawaii.

PubMed

Geller, Karly S; Nigg, Claudio R; Ollberding, Nicholas J; Motl, Robert W; Horwath, Caroline; Dishman, Rodney K

2015-03-01

Examine associations between physical activity (PA) and spatial accessibility to environmental PA resources in Hawaii. Metabolic equivalents (METs) of mild, moderate, and strenuous PA were compared for accessibility with environmental PA resources within a population-based sample of Hawaiian adults (n = 381). Multiple linear regression estimated differences in PA levels for residing further from a PA resource or residing in an area with a greater number of resources. No associations were found in the total sample. Analyses within subsamples stratified by ethnicity revealed that greater spatial accessibility to a PA resource was positively associated with strenuous PA among Caucasians (P = .04) but negatively associated with moderate PA among Native Hawaiians (P = .00). The lack of association in the total sample may be a consequence of Hawaii's unique environment. Results of stratified sample analyses are unique, providing groundwork for future examinations within parallel environments and among similar ethnic groups. © 2012 APJPH.

Hookworm infections among migrant workers in Malaysia: Molecular identification of Necator americanus and Ancylostoma duodenale.

PubMed

Sahimin, Norhidayu; Lim, Yvonne Ai Lian; Douadi, Benacer; Mohd Khalid, Mohd Khairul Nizam; Wilson, John-James; Behnke, Jerzy M; Mohd Zain, Siti Nursheena

2017-09-01

Ongoing urbanisation of the working population as well as cross-border migration of workers particularly into large cities has contributed to the development and growth of urban slums. These deprived areas are conducive for the transmission of intestinal pathogens including hookworm. The aim of this study was to determine both the prevalence and species identity of hookworm infections among the migrant worker community in Malaysia. A total of 388 faecal samples were collected from migrant workers between September 2014 and August 2015, representing workers from five employment sectors: construction, manufacturing, agriculture and plantations, food services and domestic services. Faecal samples were examined by microscopy and positive samples were subjected to molecular analysis. A total of 51 samples (13.1%) were positive by microscopy for hookworm infections. A two-step PCR based method amplifying a fragment of the 28S rRNA-ITS2 region was used to identify infections by Necator americanus and Ancylostoma spp. PCR products positive for Ancylostoma spp. were sequenced bidirectionally, and sequences analysed through BLAST and phylogenetic analysis. Samples containing Ancylostoma duodenale were further characterized by amplification and sequencing a fragment of cytochrome c oxidase subunit 1 (cox1) gene. PCR amplicons were successfully obtained from 42 (82.4%) of 51 samples, with 81.0% (34 of 42) identified as Necator americanus, 16.7% (7 of 42) as Ancylostoma spp. and 2.4% (1 of 42) as mixed infections of both species. All eight Ancylostoma spp. were confirmed to be Ancylostoma duodenale and this is the first time A. duodenale was reported in Malaysia. Samples containing A. duodenale from Nepalese and Indonesian workers shared high-similarity and were distinct compared to sequences from other countries. This study highlights the prevalence of hookworm infections among migrant workers living in Malaysia. Our findings underscore the necessity of screening migrant workers for hookworm infections, particularly those working in food-related services and industries. Copyright © 2017 Elsevier B.V. All rights reserved.

Molecular method for detection of total coliforms in drinking water samples.

PubMed

Maheux, Andrée F; Boudreau, Dominique K; Bisson, Marc-Antoine; Dion-Dupont, Vanessa; Bouchard, Sébastien; Nkuranga, Martine; Bergeron, Michel G; Rodriguez, Manuel J

2014-07-01

This work demonstrates the ability of a bacterial concentration and recovery procedure combined with three different PCR assays targeting the lacZ, wecG, and 16S rRNA genes, respectively, to detect the presence of total coliforms in 100-ml samples of potable water (presence/absence test). PCR assays were first compared to the culture-based Colilert and MI agar methods to determine their ability to detect 147 coliform strains representing 76 species of Enterobacteriaceae encountered in fecal and environmental settings. Results showed that 86 (58.5%) and 109 (74.1%) strains yielded a positive signal with Colilert and MI agar methods, respectively, whereas the lacZ, wecG, and 16S rRNA PCR assays detected 133 (90.5%), 111 (75.5%), and 146 (99.3%) of the 147 total coliform strains tested. These assays were then assessed by testing 122 well water samples collected in the Québec City region of Canada. Results showed that 97 (79.5%) of the samples tested by culture-based methods and 95 (77.9%), 82 (67.2%), and 98 (80.3%) of samples tested using PCR-based methods contained total coliforms, respectively. Consequently, despite the high genetic variability of the total coliform group, this study demonstrated that it is possible to use molecular assays to detect total coliforms in potable water: the 16S rRNA molecular assay was shown to be as efficient as recommended culture-based methods. This assay might be used in combination with an Escherichia coli molecular assay to assess drinking water quality. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Molecular Method for Detection of Total Coliforms in Drinking Water Samples

PubMed Central

Boudreau, Dominique K.; Bisson, Marc-Antoine; Dion-Dupont, Vanessa; Bouchard, Sébastien; Nkuranga, Martine; Bergeron, Michel G.; Rodriguez, Manuel J.

2014-01-01

This work demonstrates the ability of a bacterial concentration and recovery procedure combined with three different PCR assays targeting the lacZ, wecG, and 16S rRNA genes, respectively, to detect the presence of total coliforms in 100-ml samples of potable water (presence/absence test). PCR assays were first compared to the culture-based Colilert and MI agar methods to determine their ability to detect 147 coliform strains representing 76 species of Enterobacteriaceae encountered in fecal and environmental settings. Results showed that 86 (58.5%) and 109 (74.1%) strains yielded a positive signal with Colilert and MI agar methods, respectively, whereas the lacZ, wecG, and 16S rRNA PCR assays detected 133 (90.5%), 111 (75.5%), and 146 (99.3%) of the 147 total coliform strains tested. These assays were then assessed by testing 122 well water samples collected in the Québec City region of Canada. Results showed that 97 (79.5%) of the samples tested by culture-based methods and 95 (77.9%), 82 (67.2%), and 98 (80.3%) of samples tested using PCR-based methods contained total coliforms, respectively. Consequently, despite the high genetic variability of the total coliform group, this study demonstrated that it is possible to use molecular assays to detect total coliforms in potable water: the 16S rRNA molecular assay was shown to be as efficient as recommended culture-based methods. This assay might be used in combination with an Escherichia coli molecular assay to assess drinking water quality. PMID:24771030

An integrated multiple-analyte pharmacokinetic model to characterize trastuzumab emtansine (T-DM1) clearance pathways and to evaluate reduced pharmacokinetic sampling in patients with HER2-positive metastatic breast cancer.

PubMed

Lu, Dan; Joshi, Amita; Wang, Bei; Olsen, Steve; Yi, Joo-Hee; Krop, Ian E; Burris, Howard A; Girish, Sandhya

2013-08-01

Trastuzumab emtansine (T-DM1) is an antibody-drug conjugate recently approved by the US Food and Drug Administration for the treatment of human epidermal growth factor receptor 2 (HER2)-positive metastatic breast cancer previously treated with trastuzumab and taxane chemotherapy. It comprises the microtubule inhibitory cytotoxic agent DM1 conjugated to the HER2-targeted humanized monoclonal antibody trastuzumab via a stable linker. To characterize the pharmacokinetics of T-DM1 in patients with metastatic breast cancer, concentrations of multiple analytes were quantified, including serum concentrations of T-DM1 conjugate and total trastuzumab (the sum of conjugated and unconjugated trastuzumab), as well as plasma concentrations of DM1. The clearance of T-DM1 conjugate is approximately 2 to 3 times faster than its parent antibody, trastuzumab. However, the clearance pathways accounting for this faster clearance rate are unclear. An integrated population pharmacokinetic model that simultaneously fits the pharmacokinetics of T-DM1 conjugate and total trastuzumab can help to elucidate the clearance pathways of T-DM1. The model can also be used to predict total trastuzumab pharmacokinetic profiles based on T-DM1 conjugate pharmacokinetic data and sparse total trastuzumab pharmacokinetic data, thereby reducing the frequency of pharmacokinetic sampling. T-DM1 conjugate and total trastuzumab serum concentration data, including baseline trastuzumab concentrations prior to T-DM1 treatment, from phase I and II studies were used to develop this integrated population pharmacokinetic model. Based on a hypothetical T-DM1 catabolism scheme, two-compartment models for T-DM1 conjugate and trastuzumab were integrated by assuming a one-step deconjugation clearance from T-DM1 conjugate to trastuzumab. The ability of the model to predict the total trastuzumab pharmacokinetic profile based on T-DM1 conjugate pharmacokinetics and various sampling schemes of total trastuzumab pharmacokinetics was assessed to evaluate total trastuzumab sampling schemes. The final model reflects a simplified catabolism scheme of T-DM1, suggesting that T-DM1 clearance pathways include both deconjugation and proteolytic degradation. The model fits T-DM1 conjugate and total trastuzumab pharmacokinetic data simultaneously. The deconjugation clearance of T-DM1 was estimated to be ~0.4 L/day. Proteolytic degradation clearances for T-DM1 and trastuzumab were similar (~0.3 L/day). This model accurately predicts total trastuzumab pharmacokinetic profiles based on T-DM1 conjugate pharmacokinetic data and sparse total trastuzumab pharmacokinetic data sampled at preinfusion and end of infusion in cycle 1, and in one additional steady state cycle. This semi-mechanistic integrated model links T-DM1 conjugate and total trastuzumab pharmacokinetic data, and supports the inclusion of both proteolytic degradation and deconjugation as clearance pathways in the hypothetical T-DM1 catabolism scheme. The model attributes a faster T-DM1 conjugate clearance versus that of trastuzumab to the presence of a deconjugation process and suggests a similar proteolytic clearance of T-DM1 and trastuzumab. Based on the model and T-DM1 conjugate pharmacokinetic data, a sparse pharmacokinetic sampling scheme for total trastuzumab provides an entire pharmacokinetic profile with similar predictive accuracy to that of a dense pharmacokinetic sampling scheme.

Specific detection of cultivable Helicobacter pylori cells from wastewater treatment plants.

PubMed

Moreno, Yolanda; Ferrús, M Antonía

2012-10-01

Helicobacter pylori is present in surface water and wastewater, and biofilms in drinking water systems have been reported as possible reservoirs of H. pylori. However, its ability to survive in an infectious state in the environment is hindered because it rapidly loses its cultivability. The aim of this study was to determine the presence of cultivable and therefore viable H. pylori in wastewater treatment plants to understand the role of wastewater in the pathogen's transmission. A modified filter technique was used to obtain a positive H. pylori culture, and specific detection of this pathogen was achieved with FISH and PCR techniques. A total of six positive H. pylori cultures were obtained from the water samples, and molecular techniques positively identified H. pylori in 21 culture-negative samples. The combination of a culturing procedure after sample filtration followed by the application of a molecular method, such as PCR or FISH, provides a specific tool for the detection, identification, and direct visualization of cultivable and therefore viable H. pylori cells from complex mixed communities such as water samples. © 2012 Blackwell Publishing Ltd.

Prevalence and molecular characterization of Giardia duodenalis in dogs in Aydin, Turkey.

PubMed

Gultekin, Mehmet; Ural, Kerem; Aysul, Nuran; Ayan, Adnan; Balikci, Canberk; Akyildiz, Gurkan

2017-06-01

The purpose of the present study was to investigate the prevalence and molecular characterization of Giardia duodenalis among dogs in Aydin, Turkey. A total of 473 faecal samples from dogs were collected. The overall prevalence of G. duodenalis was 18.8%. Higher infection rates were observed in dogs younger than three months old, from shelters, and with diarrhoea. Faecal samples of 89 dogs, diagnosed Giardia-positive by microscopy, were found positive by nested PCR. The β-giardin nested PCR assay revealed assemblage B in all samples (100%), whereas 38 of the samples were mixed with assemblage A (42%). Sequence analysis of isolates indicated sub-genotypes A3 and B4 which have been previously detected in human isolates from Turkey. The results of the present study indicated the relatively high prevalence of giardiasis and the presence of the zoonotic sub-genotypes suggesting the important role of dogs as potential reservoirs of human infections.

Detection of antibodies against hepatitis A in blood spots dried on filter paper. Is this a reliable method for epidemiological studies?

PubMed Central

Gil, A.; González, A.; Dal-Ré, R.; Dominguez, V.; Astasio, P.; Aguilar, L.

1997-01-01

Diluted dried blood drops on filter paper were compared with serum samples as a specimen source for qualitative anti-HAV antibody determination by ELISA. A total of 298 serum samples and dried blood drops were collected from a population of healthy adolescents (15.3 +/- 1.2 years old). The prevalence of anti-HAV antibody obtained by testing serum samples was 7.7% (95% CI:4.8 10.1). Compared with serum sampling the sensitivity and specificity of diluted dried blood drops were 91.3 and 99.3%. The positive and negative predictive values were 91.3 and 99.3%, respectively, and the likelihood ratios of positive and negative results were 91 and 0.09. It is proposed that this test represents a reliable procedure for anti-HAV antibody testing. PMID:9129596

Comparison of DOT-ELISA and Standard-ELISA for Detection of the Vibrio cholerae Toxin in Culture Supernatants of Bacteria Isolated from Human and Environmental Samples.

PubMed

Meza-Lucas, Antonio; Pérez-Villagómez, María-Fernanda; Martínez-López, José-Patricio; García-Rodea, Ricardo; Martínez-Castelán, María-Guadalupe; Escobar-Gutiérrez, Alejandro; de-la-Rosa-Arana, Jorge-Luis; Villanueva-Zamudio, Altagracia

2016-09-01

A comparison of DOT-ELISA and Standard-ELISA was made for detection of Vibrio cholerae toxin in culture supernatants of bacteria isolated from human and environmental samples. A total of 293 supernatants were tested in a double blind assay. A correlation of 100 % was obtained between both techniques. The cholera toxin was found in 20 Inaba and 3 Ogawa strains. Positive samples were from seafood (17 samples), potable water (1 sample) and sewage (5 samples). The DOT-ELISA was useful as the standard-ELISA to confirm the presence of cholera toxin in the environmental samples.

Coxiellosis in domestic livestock of Puducherry and Tamil Nadu: Detection of Coxiella burnetii DNA by polymerase chain reaction in slaughtered ruminants

PubMed Central

Pradeep, Jothimani; Stephen, Selvaraj; Pooja, Pratheesh; Akshayavardhini, Anbalagan; Sangeetha, Balakrishnan; Antony, Prabakar Xavier

2017-01-01

Background and Aim:: In the course of our Indian Council of Medical Research project on coxiellosis in Puducherry and Tamil Nadu, 5.64% goat, 1.85% sheep, 1.06% buffaloes, and 0.97% cattle were positive for Coxiella burnetii antibodies by enzyme linked immunosorbent assay kit (IDEXX, Liebefeld, Switzerland). In this preliminary study, we have proceeded to look for C. burnetii DNA in those antibody positive specimens employing an imported commercial C. burnetii polymerase chain reaction (PCR) kit. Materials and Methods:: Blood samples were collected during slaughtering. All 15 blood samples of antibody positive ruminants and three antibody negative samples were subjected to conventional Trans-PCR assay with a commercial PCR kit (Genekam Biotechnology AG, Duisburg, Germany). An in-house Trans-PCR was included in the study for comparison. Results:: A total of 15 antibody positive and three antibody-negative serum samples belonging to 11 goat, 4 sheep, 1 cattle, and 2 buffaloes were tested in duplicate for the presence of C. burnetii DNA by the commercial agar gel PCR kit and an in-house Trans-PCR. Only one buffalo serum sample was positive for C. burnetii with a band at 243 bp in in-house Trans-PCR. Discussion:: Seropositivity for C. burnetii need not necessarily translate into infectivity status of the animal. Conversely, seronegative ruminants can shed C. burnetii. Rapid disintegration of C. burnetii DNA during the storage period is an important impediment in QF-PCR research. This is the first time the performance of this commercial PCR kit is being validated in India. Conclusion:: Commercial PCR kit, Genekam did not identify any positive sample, probably because it targeted a larger amplicon of 687 bp. PMID:28717320

Large-Scale Evaluation of the Immuno-Mycologics Lateral Flow and Enzyme-Linked Immunoassays for Detection of Cryptococcal Antigen in Serum and Cerebrospinal Fluid

PubMed Central

Hansen, Jessica; Slechta, E. Susan; Gates-Hollingsworth, Marcellene A.; Neary, Brandon; Barker, Adam P.; Bauman, Sean; Kozel, Thomas R.

2013-01-01

Cryptococcosis is a systemic infection caused by the pathogenic yeasts Cryptococcus neoformans and C. gattii. Detection of cryptococcal capsular antigen (CrAg) in serum and cerebrospinal fluid (CSF) plays an important diagnostic role. We prospectively compared the new Immuno-Mycologics Inc. (IMMY) lateral flow assay (LFA) and enzyme immunoassay (EIA) to our current CrAg test (Premier EIA; Meridian Bioscience Inc.). Discordant samples were retested with the latex-Cryptococcus antigen test (IMMY) and using serotype-specific monoclonal antibodies (MAbs). A total of 589 serum and 411 CSF specimens were tested in parallel. Qualitative agreement across assays was 97.7%. In all, 56 (41 serum and 15 CSF) samples were positive and 921 (527 serum and 394 CSF) samples were negative by all three assays. The 23 discrepant specimens were all Meridian EIA negative. Of 23 discordant specimens, 20 (87.0%) were positive by both the IMMY LFA and EIA, 2 were LFA positive only, and 1 was EIA positive only. Eleven discrepant specimens had adequate volume for latex agglutination (LA) testing; 8 were LA positive, and 3 were LA negative. LA-negative samples (2 CSF samples and 1 serum) had low IMMY LFA/EIA titers (≤1:10). Serotype-specific MAb analysis of the LA-positive samples suggested that these specimens contained CrAg epitopes similar to those of serotype C strains. In conclusion, the IMMY assays showed excellent overall concordance with the Meridian EIA. Assay performance differences were related to issues of analytic sensitivity and possible serotype bias. Incomplete access to patient-level data combined with low specimen volumes limited our ability to fully resolve discrepant results. PMID:23114703

Large-scale evaluation of the immuno-mycologics lateral flow and enzyme-linked immunoassays for detection of cryptococcal antigen in serum and cerebrospinal fluid.

PubMed

Hansen, Jessica; Slechta, E Susan; Gates-Hollingsworth, Marcellene A; Neary, Brandon; Barker, Adam P; Bauman, Sean; Kozel, Thomas R; Hanson, Kimberly E

2013-01-01

Cryptococcosis is a systemic infection caused by the pathogenic yeasts Cryptococcus neoformans and C. gattii. Detection of cryptococcal capsular antigen (CrAg) in serum and cerebrospinal fluid (CSF) plays an important diagnostic role. We prospectively compared the new Immuno-Mycologics Inc. (IMMY) lateral flow assay (LFA) and enzyme immunoassay (EIA) to our current CrAg test (Premier EIA; Meridian Bioscience Inc.). Discordant samples were retested with the latex-Cryptococcus antigen test (IMMY) and using serotype-specific monoclonal antibodies (MAbs). A total of 589 serum and 411 CSF specimens were tested in parallel. Qualitative agreement across assays was 97.7%. In all, 56 (41 serum and 15 CSF) samples were positive and 921 (527 serum and 394 CSF) samples were negative by all three assays. The 23 discrepant specimens were all Meridian EIA negative. Of 23 discordant specimens, 20 (87.0%) were positive by both the IMMY LFA and EIA, 2 were LFA positive only, and 1 was EIA positive only. Eleven discrepant specimens had adequate volume for latex agglutination (LA) testing; 8 were LA positive, and 3 were LA negative. LA-negative samples (2 CSF samples and 1 serum) had low IMMY LFA/EIA titers (≤1:10). Serotype-specific MAb analysis of the LA-positive samples suggested that these specimens contained CrAg epitopes similar to those of serotype C strains. In conclusion, the IMMY assays showed excellent overall concordance with the Meridian EIA. Assay performance differences were related to issues of analytic sensitivity and possible serotype bias. Incomplete access to patient-level data combined with low specimen volumes limited our ability to fully resolve discrepant results.

Real-time polymerase chain reaction and culture in the diagnosis of invasive group B streptococcal disease in infants: a retrospective study.

PubMed

Meehan, M; Cafferkey, M; Corcoran, S; Foran, A; Hapnes, N; LeBlanc, D; McGuinness, C; Nusgen, U; O'Sullivan, N; Cunney, R; Drew, R

2015-12-01

Group B streptococcus (GBS) is a leading cause of invasive disease in infants. Accurate and rapid diagnosis is crucial to reduce morbidity and mortality. Real-time polymerase chain reaction (PCR) targeting the dltR gene was utilised for the direct detection of GBS DNA in blood and cerebrospinal fluid (CSF) from infants at an Irish maternity hospital. A retrospective review of laboratory and patient records during the period 2011-2013 was performed in order to evaluate PCR and culture for the diagnosis of invasive GBS disease. A total of 3570 blood and 189 CSF samples from 3510 infants had corresponding culture and PCR results. Culture and PCR exhibited concordance in 3526 GBS-negative samples and 13 (25%) GBS-positive samples (n = 53). Six (11%) and 34 (64%) GBS-positive samples were positive only in culture or PCR, respectively. Culture and PCR identified more GBS-positive infants (n = 47) than PCR (n = 43) or culture (n = 16) alone. Using culture as the reference standard, the sensitivity, specificity, and positive and negative predictive values for PCR on blood samples were 71.4%, 99.2%, 25% and 99.9%, and for CSF samples, they were 60%, 97.8%, 42.9% and 98.9%, respectively. The sensitivity and positive predictive values were improved (blood: 84.6% and 55%; CSF: 77.8% and 100%, respectively) when maternal risk factors and other laboratory test results were considered. The findings in this study recommend the use of direct GBS real-time PCR for the diagnosis of GBS infection in infants with a clinical suspicion of invasive disease and as a complement to culture, but should be interpreted in the light of other laboratory and clinical findings.

Intrinsic religiousness and spirituality as predictors of mental health and positive psychological functioning in Latter-Day Saint adolescents and young adults.

PubMed

Sanders, Peter W; Allen, G E Kawika; Fischer, Lane; Richards, P Scott; Morgan, David T; Potts, Richard W

2015-06-01

We investigated the relationships between religiousness and spirituality and various indicators of mental health and positive psychosocial functioning in three separate samples of college students. A total of 898 students at Brigham Young University participated in the three studies. The students ranged in age from 17 to 26 years old, with the average age of 20.9 across all three samples. Our results indicate that intrinsic religiousness, spiritual maturity, and self-transcendence were significantly predictive of better mental health and positive functioning, including lower levels of depression, anxiety, and obsessive-compulsiveness, and higher levels of global self-esteem, identity integration, moral self-approval, and meaning in life. Intrinsic religiousness was not predictive of shame, perfectionism, and eating disorder symptoms. These findings are consistent with many prior studies that have found religiousness and spirituality to be positively associated with better mental health and positive psychosocial functioning in adolescents and young adults.

Kidney and urinary tract polyomavirus infection and distribution: molecular biology investigation of 10 consecutive autopsies.

PubMed

Boldorini, Renzo; Veggiani, Claudia; Barco, Diana; Monga, Guido

2005-01-01

Distinct human polyomavirus genotypes cause different diseases in patients with renal transplants: BK virus (BKV) causes tubulointerstitial nephritis and ureteral stenosis, whereas both JC virus (JCV) and BKV are responsible for hemorrhagic cystitis. These findings could result from a selective infection of kidney and urinary tract segments by JCV or BKV. To verify this hypothesis, 10 complete, unselected, consecutive autopsies from 9 immunocompetent patients and 1 patient affected by acquired immunodeficiency syndrome were investigated. Samples from kidneys (n = 80), renal pelvis (n = 20), ureter (n = 40), and urinary bladder (n = 30) obtained from 10 consecutive autopsies were investigated by means of multiplex nested polymerase chain reaction to detect polyomavirus DNA and to distinguish different species of the Polyomavirus genus. In situ hybridization and immunohistochemistry were also carried out to define the viral status of the infected tissues. Polyomavirus DNA was detected in all of the subjects (positive samples ranging from 2 to 7 samples), for a total of 43 of 170 samples (25.3%), distributed as follows: urinary bladder (10/30, 33%), renal pelvis (6/20, 30%), ureter (10/40, 25%), and kidney tissue (17/80, 21%). We found that JCV was most frequently detected overall (23/43 samples, 53.5%) and was also detected most frequently within the kidney (8/17 positive samples, 47%), the renal pelvis (5/6 positive samples, 70%), and the ureter (7/10 positive samples, 70%), whereas BKV was found in 14 samples (32.5%), and it was the prevailing genotype in urinary bladder (6/10 positive samples, 60%). Coinfection of BKV-JCV was found in 6 samples (14%). Immunohistochemistry and in situ hybridization returned negative results. The viruses JCV and BKV latently persist randomly in kidney and urinary tract. Distinct diseases induced by them could be related more closely to molecular viral rearrangements than to the topographic distribution of latent viruses.

[Avian influenza virus in various environments and risk factors for the contamination of live poultry markets during winter and spring season in Zhejiang province].

PubMed

Wang, Xiaoxiao; Cheng, Wei; Yu, Zhao; Mao, Haiyan; Chen, Enfu

2016-03-01

To evaluate the prevalence of avian influenza virus in various environment and the influence factors for subtype H7 prevalence in live poultry markets. We collected environmental samples from various environments across 11 cities of Zhejiang province between October 2014 and March 2015. Cage surface swabs, chopping board surface swabs, feces, water for cleaning, drinking water and swabs of other surfaces were collected. A total of 6 457 samples were collected, including 4 487 samples from poultry markets, 820 samples from poultry farms, 715 samples from backyard poultry pens, 118 samples from poultry processing factories, 118 samples from wild bird habitats and 86 samples from other sites. The chi-squared test was used to compare virus prevalence among sample types, sites types, and poultry markets types. Binary logistic regression was used to analyze factors on H7 subtype prevalence in poultry markets. Of 6 457 samples, 32.54% (2 101) samples were positive for avian influenza, with 3.67% (237) positive for H5 subtype, 12.02%(776) positive for H7 subtype, 11.77%(760) positive for H9 subtype. Of 237 live poultry markets, 33.8% (80) were positive for H7 subtype. The prevalence of influenza A in poultry processing factories was the highest at 43.72% (101/231) (χ(2)=737.80, P<0.001). Poultry markets were contaminated most seriously by subtype H5/H7/H9 with the prevalence of 27.55% (1 236/4 487) (χ(2)=436.37, P<0.001). Compared with markets with 1 type of poultry, OR was 4.58 (95%CI: 1.63-12.87) for markets with ≥2 types of poultry. Live poultry markets and poultry processing factories were contaminated most seriously by avian influenza. The types of poultry might be the factor which influenced the subtype H7 prevalence in poultry markets.

Avian influenza H9N2 seroprevalence among poultry workers in Pune, India, 2010.

PubMed

Pawar, Shailesh D; Tandale, Babasaheb V; Raut, Chandrashekhar G; Parkhi, Saurabh S; Barde, Tanaji D; Gurav, Yogesh K; Kode, Sadhana S; Mishra, Akhilesh C

2012-01-01

Avian influenza (AI) H9N2 has been reported from poultry in India. A seroepidemiological study was undertaken among poultry workers to understand the prevalence of antibodies against AI H9N2 in Pune, Maharashtra, India. A total of 338 poultry workers were sampled. Serum samples were tested for presence of antibodies against AI H9N2 virus by hemagglutination inhibition (HI) and microneutralization (MN) assays. A total of 249 baseline sera from general population from Pune were tested for antibodies against AI H9N2 and were negative by HI assay using ≥40 cut-off antibody titre. Overall 21 subjects (21/338 = 6.2%) were positive for antibodies against AI H9N2 by either HI or MN assays using ≥40 cut-off antibody titre. A total of 4.7% and 3.8% poultry workers were positive for antibodies against AI H9N2 by HI and MN assay respectively using 40 as cut-off antibody titre. This is the first report of seroprevalence of antibodies against AI H9N2 among poultry workers in India.

[Effect of afforestation modes on soil microbial community and nitrogen functional genes in Hippophae rhamnoides plantation].

PubMed

Yang, Dan; Yu, Xuan; Liu, Xu; Liu, Jin-liana; Zhang, Shun-xiang; Yu, Ze-qun

2015-12-01

The study aimed to assess the effect of different afforestation modes on microbial composition and nitrogen functional genes in soil. Soil samples from a pure Hippophae rhamnoides stand (SS) and three mixed stands, namely, H. rhamnoides and Pinus tabuliformis (SY), H. rhamnoides and Platycladus orientalis (SB), H. rhamnoides and Robinia pseucdoacacia (SC) were selected. The results showed that the total PLFA (TPLFA), bacterial PLFA, gram positive bacterial PLFA (G⁺PLFA) were significantly higher in soil samples from other three stands than those of the pure one. However, no significant difference was found for fungal PLFA among them. The abundance of nifH, amoA, nirK and narG genes were higher in SY and SC than in SS. The TPLFA, G⁺PLFA, gram negative bacterial PLFA (G⁻PLFA), and all of the detected gene abundance were significantly and positively correlated with soil pH, total organic carbon, total nitrogen, ammonium nitrogen and available potassium. Afforestation modes affected indirectly soil microbial composition and functional genes through soil properties. Mixing P. tabuliformis or P. orientalis with H. rhamnoides might be suitable afforestation modes, which might improve soil quality.

Attenuated total reflectance spectroscopy of plant leaves: A tool for ecological and botanical studies

USGS Publications Warehouse

Ribeiro da Luz, B.

2006-01-01

??? Attenuated total reflectance (ATR) spectra of plant leaves display complex absorption features related to organic constituents of leaf surfaces. The spectra can be recorded rapidly, both in the field and in the laboratory, without special sample preparation. ??? This paper explores sources of ATR spectral variation in leaves, including compositional, positional and temporal variations. Interspecific variations are also examined, including the use of ATR spectra as a tool for species identification. ??? Positional spectral variations generally reflected the abundance of cutin and the epicuticular wax thickness and composition. For example, leaves exposed to full sunlight commonly showed more prominent cutin- and wax-related absorption features compared with shaded leaves. Adaxial vs. abaxial leaf surfaces displayed spectral variations reflecting differences in trichome abundance and wax composition. Mature vs. young leaves showed changes in absorption band position and intensity related to cutin, polysaccharide, and possibly amorphous silica development on and near the leaf surfaces. ??? Provided that similar samples are compared (e.g. adaxial surfaces of mature, sun-exposed leaves) same-species individuals display practically identical ATR spectra. Using spectral matching procedures to analyze an ATR database containing 117 individuals, including 32 different tree species, 83% of the individuals were correctly identified. ?? The Authors (2006).

77 FR 58091 - Risk-Based Sampling of Beef Manufacturing Trimmings for Escherichia coli (E. coli) O157:H7 and...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-19

... (total volume CY 2007- 2011: 1,268,625,000 pounds) has the highest E. coli O157:H7 percent positive. The... that the percent-positive rate in the high- prevalence season (now considered to be May through October, rather than April through September, on the basis of FSIS data) is about 2 times as high as it is the...

Monitoring of Microscopic Filamentous Fungi in Indoor Air of Transplant Unit.

PubMed

Holý, Ondřej; Matoušková, Ivanka; Kubátová, Alena; Hamal, Petr; Svobodová, Lucie; Jurásková, Eva; Raida, Luděk

2015-12-01

The aim of the study was to control the microbial contamination of indoor air monitored monthly at the Transplant Unit of the University Hospital Olomouc from August 2010 to July 2011. The unit is equipped with a three-stage air filtration system with HEPA filters. The MAS-100 air sampler (Merck, GER) was used. Twenty locations were singled out for the purposes of collecting a total of 720 samplings of the indoor air. Swabs of the HVAC diffusers at the sampling locations were always carried out after the sampling of the indoor air. In total, 480 samples of the indoor air were taken for Sabouraud chloramphenicol agar. In 11 cases (2.29%) the cultivation verified the presence of microscopic filamentous fungi. Only two cases involved the sanitary facilities of a patient isolation box; the other positive findings were from the facilities. The most frequent established genus was Aspergillus spp. (4x), followed by Trichoderma spp. (2x) and Penicillium spp. (2x), Paecilomyces spp., Eurotium spp., and Chrysonilia spp. (1x each). In 2 cases the cultivation established sterile aerial mycelium, unfortunately no further identification was possible. A total of 726 swabs of HVAC diffusers were collected (2 positive-0.28%). The study results demonstrated the efficacy of the HVAC equipment. With the continuing increase in the number of severely immunocompromised patients, hospitals are faced with the growing problem of invasive aspergillosis and other opportunistic infections. Preventive monitoring of microbial air contaminants is of major importance for the control of invasive aspergillosis. Copyright© by the National Institute of Public Health, Prague 2015.

Background concentrations of metals in soils from selected regions in the State of Washington

USGS Publications Warehouse

Ames, K.C.; Prych, E.A.

1995-01-01

Soil samples from 60 sites in the State of Washington were collected and analyzed to determine the magnitude and variability of background concen- trations of metals in soils of the State. Samples were collected in areas that were relatively undisturbed by human activity from the most pre- dominant soils in 12 different regions that are representative of large areas of Washington State. Concentrations of metals were determined by five different laboratory methods. Concentrations of mercury and nickel determined by both the total and total-recoverable methods displayed the greatest variability, followed by chromium and copper determined by the total-recoverable method. Concentrations of other metals, such as aluminum and barium determined by the total method, varied less. Most metals concentrations were found to be more nearly log-normally than normally distributed. Total metals concentrations were not significantly different among the different regions. However, total-recoverable metals concentrations were not as similar among different regions. Cluster analysis revealed that sampling sites in three regions encompassing the Puget Sound could be regrouped to form two new regions and sites in three regions in south-central and southeastern Washington State could also be regrouped into two new regions. Concentrations for 7 of 11 total-recoverable metals correlated with total metals concentrations. Concen- trations of six total metals also correlated positively with organic carbon. Total-recoverable metals concentrations did not correlate with either organic carbon or particle size. Concentrations of metals determined by the leaching methods did not correlate with total or total-recoverable metals concentrations, nor did they correlate with organic carbon or particle size.

Serosurvey for antibodies to malignant catarrhal fever-associated viruses in free-living and captive cervids in Germany.

PubMed

Frölich, K; Li, H; Müller-Doblies, U

1998-10-01

A total of 486 serum samples collected from several species of both free-living and captive cervids in Germany was examined for antibodies against malignant catarrhal fever (MCF)-associated viruses (MCFV) by a competitive-inhibition enzyme-linked immunosorbent assay (CI-ELISA). Eleven (2%) of these samples were positive for antibodies against MCFV. Among 157 serum samples collected from 16 different species of captive deer including four (7%) of 54 fallow deer and one (7%) of 14 sika deer (Cervus nippon) were seropositive. Among 329 samples from three species of free-ranging deer, including 253 roe deer (Capreolus capreolus), 22 red deer (Cervus elaphus) and 54 fallow deer (Cervus dama), only fallow deer were antibody-positive. Of the 25 fallow deer samples collected between 1990 and 1993, four (16%) were seropositive. Among 29 free-ranging fallow deer samples collected in the hunting period 1996-1997, antibodies to MCFV were detected in two (7%) of these sera. All of these fallow deer samples were collected from a circumscribed area in northern Germany. In the same area a high seroprevalence (72%) to MCFV was observed in domestic sheep (n = 50). Among 20 sheep samples (buffy coat) and 15 fallow deer samples (spleen or lymph nodes) examined for ovine herpesvirus 2 (OvHV-2) by PCR, all 20 sheep samples examined were OvHV-2 positive, but all of the 15 fallow deer samples, including seven seropositive deer, were OvHV-2 negative.

Profile of microflora of the posterior intestine of Chinook salmon before, during, and after administration of rations with and without erythromycin

USGS Publications Warehouse

Moffitt, C.M.; Mobin, S.M.A.

2006-01-01

We describe the resident heterotrophic aerobic microflora of the salmonid posterior intestine before, during, and after the administration of rations with erythromycin in a hatchery raceway environment. We compare the profiles of medicated Chinook salmon Oncorhynchus tshawytscha with those of control fish that were not fed erythromycin. The combined counts of bacteria and yeasts per gram of fish intestine originating from four upstream raceways ranged from 3.0 ?? 102 to 9.6 ?? 105 colony-forming units (CFU) over the study period. Yeasts were commonly identified in the gut, and abundances ranged from 0% to more than 80% of the CFU. Erythromycin therapy decreased the total microbial population and altered the bacterial diversity in the gut during treatment. The intestinal microbial populations in fish medicated with erythromycin increased rapidly after treatment ceased, and by 25 d after treatment the CFU were similar in samples from both medicated and control fish populations. Of 325 isolates from fish selected for biochemical profiles, we identified a total of eight gram-positive and eight gram-negative genera. Bacillus spp. were common throughout sampling and were identified in samples of fish feed. Erythromycin-resistant, gram-positive bacteria were observed throughout the sampling in medicated and control fish. We identified seven gram-positive and two gram-negative genera in 74 selected isolates from control and erythromycin feeds. Our studies suggest that the aerobic microflora of the posterior intestine varies over time, and it is likely that few resistant genera of concern to human health are present.

First detection and characterization of Salmonella spp. in poultry and swine raised in backyard production systems in central Chile.

PubMed

Alegria-Moran, R; Rivera, D; Toledo, V; Moreno-Switt, A I; Hamilton-West, C

2017-11-01

Little is known about Salmonella serovars circulating in backyard poultry and swine populations worldwide. Backyard production systems (BPS) that raise swine and/or poultry are distributed across Chile, but are more heavily concentrated in central Chile, where industrialized systems are in close contact with BPS. This study aims to detect and identify circulating Salmonella serovars in poultry and swine raised in BPS. Bacteriological Salmonella isolation was carried out for 1744 samples collected from 329 BPS in central Chile. Faecal samples were taken from swine, poultry, geese, ducks, turkeys and peacocks, as well as environmental faecal samples. Confirmation of Salmonella spp. was performed using invA-polymerase chain reaction (PCR). Identification of serovars was carried out using a molecular serotyping approach, where serogroups were confirmed by a multiplex PCR of Salmonella serogroup genes for five Salmonella O antigens (i.e., D, B, C1, C2-C3, and E1), along with two PCR amplifications, followed by sequencing of fliC and fljB genes. A total of 25 samples (1·4% of total samples) from 15 BPS (4·6 % of total sampled BPS) were found positive for Salmonella. Positive samples were found in poultry (chickens and ducks), swine and environmental sources. Molecular prediction of serovars on Salmonella isolated showed 52·0% of S. Typhimurium, 16·0% of S. Infantis, 16·0% S. Enteritidis, 8·0% S. Hadar, 4·0% S. Tennessee and 4·0% S. Kentucky. Poor biosecurity measures were found on sampled BPS, where a high percentage of mixed confinement systems (72·8%); and almost half of the sampled BPS with improper management of infected mortalities (e.g. selling the carcasses of infected animals for consumption). Number of birds other than chickens (P = 0·014; OR = 1·04; IC (95%) = 1·01-1·07), mixed productive objective (P = 0·030; OR = 5·35; IC (95%) = 1·24-27·59) and mixed animal replacement origin (P = 0017; OR = 5·19; IC (95%) = 1·35-20·47) were detected as risk factors for BPS positivity to Salmonella spp. This is the first evidence of serovars of Salmonella spp. circulating in BPS from central Chile. Detected serovars have been linked to human and animal clinical outbreaks worldwide and in Chile, highlighting the importance of BPS on the control and dissemination of Salmonella serovars potentially hazardous to public health.

Methylation of Werner syndrome protein is associated with the occurrence and development of invasive meningioma via the regulation of Myc and p53 expression.

PubMed

Li, Puxian; Hao, Shuyu; Bi, Zhiyong; Zhang, Junting; Wu, Zhen; Ren, Xiaohui

2015-08-01

The aim of the present study was to investigate the positive rate of Werner syndrome protein (WRN) methylation in meningioma patients, and further assess the association between WRN methylation and the occurrence of meningioma. A total of 56 consecutive meningioma patients and 26 healthy individuals were enrolled in the study. A methylation-specific polymerase chain reaction assay was performed to detect the positive rate of WRN methylation in the peripheral blood and tissue samples collected from the recruited subjects. In addition, western blot analysis was performed to determine the protein expression levels of WRN, Myc and p53 in the peripheral blood and tissue samples. The positive rate of WRN methylation in the peripheral blood of the meningioma group was increased when compared with the control group (P<0.05). In addition, the protein expression levels of WRN were significantly decreased in the peripheral blood and tissue samples collected from the individuals with a positive WRN methylation status (P<0.05), as compared with the samples without WRN methylation. Furthermore, the protein expression levels of Myc and p53 were increased in the peripheral blood and tissue samples that exhibited positive WRN methylation when compared with those without WRN methylation (P<0.05). Therefore, WRN methylation was demonstrated to be associated with the occurrence and development of invasive meningioma, possibly through the regulation of Myc and p53 expression.

Speciation and monitoring test for inorganic arsenic in white rice flour.

PubMed

Narukawa, Tomohiro; Hioki, Akiharu; Chiba, Koichi

2012-02-01

A monitoring test for arsenic species in white rice flour was developed and applied to flours made from 20 samples of polished rice collected from locations all over Japan. The arsenic species in white rice flour made from five samples each of four types of rice were analyzed by HPLC-ICP-MS after a heat-assisted aqueous extraction. The total arsenic and major and minor element concentrations in the white rice flours were measured by ICP-MS and ICP-OES after microwave-assisted digestion. 91 ± 1% of the arsenic in the flours was extractable. Concentrations of arsenite [As(III)], arsenate [As(V)], and dimethylarsinic acid (DMAA) were closely positively correlated with the total arsenic concentrations. The total arsenic concentration in flours made from rice collected around Japan was 0.15 ± 0.07 mg kg(-1) (highest, 0.32 mg kg(-1)), which is very low. It was thus confirmed that the white rice flour samples collected in this experiment were not suffered from noticeable As contamination.

Assessment of groundwater quality of Ballia district, Uttar Pradesh, India, with reference to arsenic contamination using multivariate statistical analysis

NASA Astrophysics Data System (ADS)

Singh, Asha Lata; Singh, Vipin Kumar

2018-06-01

A total of 22 water quality parameters were selected for the analysis of groundwater samples with reference to arsenic contamination. Samples were collected in the pre-monsoon and monsoon seasons of the year 2013. The maximum arsenic concentration in both the pre-monsoon and monsoon seasons was approximately the same, i.e., the maximum arsenic concentration being 75.60 and 74.46 µg/L in pre-monsoon and monsoon, respectively. Out of 72 collected samples, three were below the WHO guideline value of 10 µg/L for arsenic concentration. In 95.83% of the groundwater samples, the arsenic concentration was above the permissible limit. Nickel, manganese, and chromium concentrations were above the permissible limits in nearly all samples except for chromium concentration in a few pre-monsoon samples. However, the total iron concentrations in 23 samples (31.94%) were above the permissible limit. A total of six and seven principal components (PCs) were extracted using principal component analysis during the pre-monsoon and monsoon seasons, respectively, accounting for 76.25 and 78.52% of the total variation during two consecutive seasons. Correlation statistics revealed that the arsenic concentration was positively correlated with phosphate, iron, ammonium, bicarbonate, and manganese concentrations but negatively correlated with oxidation reduction potential (ORP), sulfate concentration, electrical conductivity, and total dissolved solids concentration. The negative correlation of arsenic with ORP suggested reducing conditions prevailing in the groundwater. The trilinear Piper diagram revealed calcium and magnesium enrichment of groundwater with an abundance of chloride ions but no predominance of bicarbonate ions. Thus, the groundwater fell into Ca2+ - Mg2+ - Cl- - SO4 2- category.

The Use of Multiplex PCR to Determine the Prevalence of Enterotoxigenic Staphylococcus aureus Isolated from Raw Milk, Feta Cheese, and Hand Swabs.

PubMed

Zeinhom, Mohamed M A; Abdel-Latef, Gihan K; Jordan, Kieran

2015-12-01

Staphylococcus aureus (S. aureus) can cause mastitis in cattle and, therefore, can be present in milk. This study was undertaken to determine the prevalence of coagulase positive S. aureus and its enterotoxin genes sea, seb, and sec in isolates recovered from raw milk, feta cheese, and human hand swabs of milk and cheese handlers in Beni-Suef province, Egypt. A total of 100 samples of raw milk and 50 samples of pasteurized-milk feta cheese were collected. In addition, 50 hand swabs from milk handlers and 25 hand swabs from cheese handlers were examined for the presence of coagulase positive S. aureus. The isolates were characterized by multiplex PCR for detection of sea, seb, and sec genes, and for resistance to 5 classes of commonly used antibiotics. Twelve (12/100), 12 (6/50), and 17% (13/75) of milk, cheese, and hand swab samples, respectively, were positive for coagulase positive S. aureus. One isolate was obtained from each positive sample (31 isolates), and none contained genes for SEA or SEC production. Twenty-five percent, 33%, and 31%, respectively, of the isolates contained the genes for SEB, resulting in 3%, 4%, and 5% of samples being positive for toxin producing coagulase positive S. aureus, respectively. At least one isolate was resistant to each of the antibiotics tested. Despite the low potential for SEB production shown, preventative measures, such as maintenance of the cold-chain and good hygienic practices should be implemented to further reduce the potential risk to public health from SEB, and to reduce the spread of antimicrobial resistance. © 2015 Institute of Food Technologists®

Field comparison of real-time polymerase chain reaction and bacterial culture for identification of bovine mastitis bacteria.

PubMed

Koskinen, M T; Wellenberg, G J; Sampimon, O C; Holopainen, J; Rothkamp, A; Salmikivi, L; van Haeringen, W A; Lam, T J G M; Pyörälä, S

2010-12-01

Fast and reliable identification of the microorganisms causing mastitis is important for management of the disease and for targeting antimicrobial treatment. Methods based on PCR are being used increasingly in mastitis diagnostics. Comprehensive field comparisons of PCR and traditional milk bacteriology have not been available. The results of a PCR kit capable of detecting 11 important etiological agents of mastitis directly from milk in 4h were compared with those of conventional bacterial culture (48h). In total, 1,000 quarter milk samples were taken from cows with clinical or subclinical mastitis, or from clinically healthy quarters with low somatic cell count (SCC). Bacterial culture identified udder pathogens in 600/780 (77%) of the clinical samples, whereas PCR identified bacteria in 691/780 (89%) of the clinical samples. The PCR analysis detected major pathogens in a large number of clinical samples that were negative for the species in culture. These included 53 samples positive for Staphylococcus aureus by PCR, but negative by culture. A total of 137 samples from clinical mastitis, 5 samples from subclinical mastitis, and 1 sample from a healthy quarter were positive for 3 or more bacterial species in PCR, whereas culture identified 3 or more species in 60 samples from clinical mastitis. Culture identified a species not targeted by the PCR test in 44 samples from clinical mastitis and in 9 samples from subclinical mastitis. Low SCC samples provided a small number of positive results both in culture (4/93; 4.3%) and by PCR (7/93; 7.5%). In conclusion, the PCR kit provided several benefits over conventional culture, including speed, automated interpretation of results, and increased sensitivity. This kit holds much promise as a tool to complement traditional methods in identification of pathogens. In conventional mastitis bacteriology, a sample with 3 or more species is considered contaminated, and resampling of the cow is recommended. Further study is required to investigate how high sensitivity of PCR and its quantitative features can be applied to improve separation of relevant udder pathogens from likely contaminants in samples where multiple species are detected. Furthermore, increasing the number of species targeted by the PCR test would be advantageous. Copyright © 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Effectiveness of sampling methods employed for Acanthamoeba keratitis diagnosis by culture.

PubMed

Muiño, Laura; Rodrigo, Donoso; Villegas, Rodrigo; Romero, Pablo; Peredo, Daniel E; Vargas, Rafael A; Liempi, Daniela; Osuna, Antonio; Jercic, María Isabel

2018-06-18

This retrospective, observational study was designed to evaluate the effectiveness of the sampling methods commonly used for the collection of corneal scrapes for the diagnosis of Acanthamoeba keratitis (AK) by culture, in terms of their ability to provide a positive result. A total of 553 samples from 380 patients with suspected AK received at the Parasitology Section of the Public Health Institute of Chile, between January 2005 and December 2015, were evaluated. A logistic regression model was used to determine the correlation between the culture outcome (positive or negative) and the method for sample collection. The year of sample collection was also included in the analysis as a confounding variable. Three hundred and sixty-five samples (27%) from 122 patients (32.1%) were positive by culture. The distribution of sample types was as follows: 142 corneal scrapes collected using a modified bezel needle (a novel method developed by a team of Chilean corneologists), 176 corneal scrapes obtained using a scalpel, 50 corneal biopsies, 30 corneal swabs, and 155 non-biological materials including contact lens and its paraphernalia. Biopsy provided the highest likelihood ratio for a positive result by culture (1.89), followed by non-biological materials (1.10) and corneal scrapes obtained using a modified needle (1.00). The lowest likelihood ratio was estimated for corneal scrapes obtained using a scalpel (0.88) and cotton swabs (0.78). Apart from biopsy, optimum corneal samples for the improved diagnosis of AK can be obtained using a modified bezel needle instead of a scalpel, while cotton swabs are not recommended.

Evaluation of Bacterial Contamination as an Indicator of Viral Contamination in a Sedimentary Aquifer in Uruguay.

PubMed

Gamazo, P; Victoria, M; Schijven, J F; Alvareda, E; Tort, L F L; Ramos, J; Burutaran, L; Olivera, M; Lizasoain, A; Sapriza, G; Castells, M; Colina, R

2018-03-21

In Uruguay, groundwater is frequently used for agricultural activities, as well as for human consumption in urban and rural areas. As in many countries worldwide, drinking water microbiological quality is evaluated only according to bacteriological standards and virological analyses are not mentioned in the legislation. In this work, the incidence of human viral (Rotavirus A, Norovirus GII, and human Adenovirus) and bacterial (total and thermotolerant coliform and Pseudomonas aeruginosa) contamination in groundwater in the Salto district, Uruguay, as well as the possible correlation between these groups of microorganisms, was studied. From a total of 134 groundwater samples, 42 (32.1%) were positive for Rotavirus, only 1 (0.7%) for both Rotavirus and Adenovirus, and 96 (72.6%) samples were positive for bacterial indicators. Results also show that Rotavirus presence was not associated with changes in chemical composition of the aquifer water. Bacteriological indicators were not adequate to predict the presence of viruses in individual groundwater samples (well scale), but a deeper spatial-temporal analysis showed that they are promising candidates to assess the viral contamination degree at aquifer scale, since from the number of wells with bacterial contamination the number of wells with viral contamination could be estimated.

Comparison of the efficacy of disinfectants in automated endoscope reprocessors for colonoscopes: tertiary amine compound (Sencron2®) versus ortho-phthalaldehyde (Cidex®OPA)

PubMed Central

Seo, Hyun Il; Lee, Dae Sung; Yoon, Eun Mi; Kwon, Min-Jung; Park, Hyosoon; Jung, Yoon Suk; Park, Jung Ho; Sohn, Chong Il

2016-01-01

Background/Aims To prevent the transmission of pathogens by endoscopes, following established reprocessing guidelines is critical. An ideal reprocessing step is simple, fast, and inexpensive. Here, we evaluated and compared the efficacy and safety of two disinfectants, a tertiary amine compound (TAC) and ortho-phthalaldehyde (OPA). Methods A total of 100 colonoscopes were randomly reprocessed using two same automated endoscope reprocessors, according to disinfectant. The exposure time was 10 minutes for 0.55% OPA (Cidex® OPA, Johnson & Johnson) and 5 minutes for 4% TAC (Sencron2®, Bab Gencel Pharma & Chemical Ind. Co.). Three culture samples were obtained from each colonoscope after reprocessing. Results A total of nine samples were positive among the 300 culture samples. The positive culture rate was not statistically different between the two groups (4% for OPA and 2% for TAC, P=0.501). There were no incidents related to safety during the study period. Conclusions TAC was non-inferior in terms of reprocessing efficacy to OPA and was safe to use. Therefore, TAC seems to be a good alternative disinfectant with a relatively short exposure time and is also less expensive than OPA. PMID:27175119

Water characteristics associated with the occurrence of Legionella pneumophila in dental units.

PubMed

Zanetti, F; Stampi, S; De Luca, G; Fateh-Moghadam, P; Antonietta, M; Sabattini, B; Checchi, L

2000-02-01

This study evaluated the incidence of Legionella pneumophila in dental unit water samples and investigated how the occurrence of these bacteria may be related to some physical, chemical and bacteriological characteristics of the water. The samples were taken from the incoming tap water, oral rinsing cup, air-water syringe, ultrasonic scaler, and the turbine of 23 dental units of private and public institutions. Apart from L. pneumophila (serogroup 1 and 3) isolated in 22 out of the 101 (21.8%) water samples tested, two other species were found: L. bozemanii and L. dumoffii. The highest densities and frequency of L. pneumophila were observed in the water coming into the units and in the dental units of public institutions. A negative association between L. pneumophila and 36 degrees C and 22 degrees C heterotrophic total plate counts and other gram-negative bacteria was found. An inverse association between the concentration of L. pneumophila and water temperature was also observed. The values of pH and total hardness did not show any significant difference in the L. pneumophila-positive and -negative dental unit waters. Finally, the chemical oxygen demand (COD) and residual chlorine were found to correlate positively with L. pneumophila.

Detection of reticuloendotheliosis virus as a contaminant of fowl pox vaccines.

PubMed

Awad, A M; Abd El-Hamid, H S; Abou Rawash, A A; Ibrahim, H H

2010-11-01

This study was designed to detect reticuloendotheliosis virus (REV) as a contaminant in fowl pox vaccines. A total of 30 fowl pox vaccine samples were examined for the presence of REV using both in vitro and in vivo methods. In in vitro testing, the fowl pox vaccine samples were inoculated into chicken embryo fibroblast cultures prepared from specific-pathogen-free embryonated chicken eggs, and the cultures were examined using PCR to detect REV. In in vivo testing, each fowl pox vaccine sample was inoculated into 5-d-old specific-pathogen-free chicks, which were kept under observation for up to 12 wk postinoculation; serum samples were collected at 15, 30, and 45 d postinoculation for the detection of REV-specific antibodies using ELISA. Tissue samples were collected at 8 and 12 wk postinoculation for histopathological examination. Of the tested vaccines, only one imported vaccine sample tested positive for REV using PCR. Serum samples collected from chicks infected with the PCR-positive vaccine batch also tested positive for REV-specific antibodies using ELISA. Histopathological examination of the liver, spleen, and bursa of Fabricius demonstrated the presence of tumor cells in these organs, confirming the results obtained using PCR and ELISA, and indicating that the sample was contaminated with REV. These data clearly indicate that the screening of all commercial poultry vaccines for viruses is an important factor in assuring the biosafety of animal vaccines.

Aspergillus and aflatoxin in groundnut (Arachis hypogaea L.) and groundnut cake in Eastern Ethiopia.

PubMed

Mohammed, Abdi; Chala, Alemayehu; Dejene, Mashilla; Fininsa, Chemeda; Hoisington, David A; Sobolev, Victor S; Arias, Renee S

2016-12-01

This study was conducted to assess major Aspergillus species and aflatoxins associated with groundnut seeds and cake in Eastern Ethiopia and evaluate growers' management practices. A total of 160 groundnut seed samples from farmers' stores and 50 groundnut cake samples from cafe and restaurants were collected. Fungal isolation was done from groundnut seed samples. Aspergillus flavus was the dominant species followed by Aspergillus parasiticus. Aflatoxin analyses of groundnut seed samples were performed using ultra performance liquid chromatography; 22.5% and 41.3% of samples were positive, with total aflatoxin concentrations of 786 and 3135 ng g -1 from 2013/2014 and 2014/2015 samples, respectively. The level of specific aflatoxin concentration varied between 0.1 and 2526 ng g -1 for B 2 and B 1 , respectively. Among contaminated samples of groundnut cake, 68% exhibited aflatoxin concentration below 20 ng g -1 , while as high as 158 ng g -1 aflatoxin B 1 was recorded. The study confirms high contamination of groundnut products in East Ethiopia.

Evaluation of the Roche LightMix Gastro parasites multiplex PCR assay detecting Giardia duodenalis, Entamoeba histolytica, cryptosporidia, Dientamoeba fragilis, and Blastocystis hominis.

PubMed

Friesen, J; Fuhrmann, J; Kietzmann, H; Tannich, E; Müller, M; Ignatius, R

2018-03-23

Multiplex PCR assays offer highly sensitive and specific tools for the detection of enteric pathogens. This prospective study aimed at comparing the novel Roche LightMix Modular Assay Gastro Parasites (LMAGP) detecting Giardia duodenalis, Entamoeba histolytica, Cryptosporidium spp., Blastocystishominis, and Dientamoebafragilis with routine laboratory procedures. Stool specimens (n = 1062 from 1009 patients) were consecutively examined by LMAGP, R-Biopharm Ridascreen enzyme immunoassays (EIAs) detecting G. duodenalis or E. histolytica/dispar, and microscopy of wet mounts. Discrepant results were analysed by in-house PCR. D. fragilis or B. hominis were detected by LMAGP in 131 (14.4%) and 179 (19.9%; 16 samples positive by microscopy; p < 0.0001) of 909 samples, respectively. Of 918 samples analysed for Cryptosporidium spp., six were positive by LMAGP (three could be confirmed by Kinyoun staining and one by in-house PCR). G. duodenalis was detected by LMAGP, EIA, or microscopy in 20, 16, or 9 of 1039 stool samples, respectively; all four samples missed by EIA were confirmed by in-house PCR. In total, 938 stool samples were analysed for E. histolytica/dispar. Nine of ten EIA-positive samples were negative by LMAGP but positive by in-house PCR for E. dispar. One E. histolytica infection (positive by both LMAGP and in-house PCR) was missed by EIA and microscopy. Parasites only detected by microscopy included Enterobius vermicularis eggs (n = 3) and apathogenic amoebae (n = 27). The data call for routine use of multiplex PCR assays for the detection of enteric protozoan parasites in laboratory diagnostics. Copyright © 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Stress-Induced Subclinical Reactivation of Varicella Zoster Virus in Astronauts

NASA Technical Reports Server (NTRS)

Mehta, Satish K.; Pierson, Duane L.; Forghani, Bagher; Zerbe, Gary; Cohrs, Randall J.; Gilden, Donald H.

2003-01-01

After primary infection, varicella-zoster virus (VZV) becomes latent in ganglia. VZV reactivation occurs primarily in elderly individuals, organ transplant recipients, and patients with cancer and AIDS, correlating with a specific decline in cell-mediated immunity to VZV. VZV can also reactivate after surgical stress. To determine whether VZV can also reactivate after acute non-surgical stress, we examined total DNA extracted from 312 saliva samples of eight astronauts before, during and after space flight for VZV DNA by PCR: 112 samples were obtained 234 to 265 days before flight, 84 samples on days 2 through 13 of space flight, and 116 samples on days 1 through 15 after flight. Before space flight only one of the 112 saliva samples from a single astronaut was positive for VZV DNA. In contrast, during and after space flight, 61 of 200 (30%) saliva samples were positive in all 8 astronauts. No VZV DNA was detected in any of 88 saliva samples from 10 healthy control subjects. These data indicate that VZV can reactivate subclinically in healthy individuals after acute stress.

Progress toward the determination of correct classification rates in fire debris analysis.

PubMed

Waddell, Erin E; Song, Emma T; Rinke, Caitlin N; Williams, Mary R; Sigman, Michael E

2013-07-01

Principal components analysis (PCA), linear discriminant analysis (LDA), and quadratic discriminant analysis (QDA) were used to develop a multistep classification procedure for determining the presence of ignitable liquid residue in fire debris and assigning any ignitable liquid residue present into the classes defined under the American Society for Testing and Materials (ASTM) E 1618-10 standard method. A multistep classification procedure was tested by cross-validation based on model data sets comprised of the time-averaged mass spectra (also referred to as total ion spectra) of commercial ignitable liquids and pyrolysis products from common building materials and household furnishings (referred to simply as substrates). Fire debris samples from laboratory-scale and field test burns were also used to test the model. The optimal model's true-positive rate was 81.3% for cross-validation samples and 70.9% for fire debris samples. The false-positive rate was 9.9% for cross-validation samples and 8.9% for fire debris samples. © 2013 American Academy of Forensic Sciences.

The adjusted effect of maternal body mass index, energy and macronutrient intakes during pregnancy, and gestational weight gain on body composition of full-term neonates.

PubMed

Pereira-da-Silva, Luis; Cabo, Catarina; Moreira, Ana Catarina; Virella, Daniel; Guerra, Tatiana; Camoes, Tania; Silva, Ana Rita; Neves, Rute; Ferreira, Goncalo Cordeiro

2014-11-01

To evaluate the effect of prepregnancy body mass index (BMI), energy and macronutrient intakes during pregnancy, and gestational weight gain (GWG) on the body composition of full-term appropriate-for-gestational age neonates. This is a cross-sectional study of a systematically recruited convenience sample of mother-infant pairs. Food intake during pregnancy was assessed by food frequency questionnaire and its nutritional value by the Food Processor Plus (ESHA Research Inc, Salem, OR). Neonatal body composition was assessed both by anthropometry and air displacement plethysmography. Explanatory models for neonatal body composition were tested by multiple linear regression analysis. A total of 100 mother-infant pairs were included. Prepregnancy overweight was positively associated with offspring weight, weight/length, BMI, and fat-free mass in the whole sample; in males, it was also positively associated with midarm circumference, ponderal index, and fat mass. Higher energy intake from carbohydrate was positively associated with midarm circumference and weight/length in the whole sample. Higher GWG was positively associated with weight, length, and midarm circumference in females. Positive adjusted associations were found between both prepregnancy BMI and energy intake from carbohydrate and offspring body size in the whole sample. Positive adjusted associations were also found between prepregnancy overweight and adiposity in males, and between GWG and body size in females. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Psychometric properties of revised Thought-Action Fusion questionnaire (TAF-R) in an Iranian population.

PubMed

Pourfaraj, Majid; Mohammadi, Nourallah; Taghavi, Mohammadreza

2008-12-01

The purpose of this study is to examine the psychometric properties of Thought-Action Fusion revised scale (TAF-R; Amir, N., freshman, M., Ramsey, B., Neary, E., & Brigidi, B. (2001). Thought-action fusion in individuals with OCD symptoms. Behaviour Research and Therapy, 39, 765-776) in a sample of 565 (321 female) students of Shiraz university. The results of factor analysis with using varimax rotation yielded eight factors that explained 80% variances of total scale. These factors are labeled: moral TAF, responsibility for positive thoughts, likelihood negative events, likelihood positive events, responsibility for negative thoughts, responsibility for harm avoidance, likelihood harm avoidance and likelihood self, respectively. The reliability coefficients of total scale are calculated by two methods: internal consistency and test-retest, which were 0.81 and 0.61, respectively. Concurrent validity showed that TAF-R scores positively and significantly correlate with responsibility, guilt and obsessive-compulsive symptoms. Confirming the expectations, there were people with high obsessive-compulsive symptoms having higher TAF-R scores than those with low symptoms. Moreover, subscales-total correlations showed that the correlations between subscales were low, but subscales correlating with total score of TAF-R were moderated.

[Superficial infections caused by Microsporum canis in humans and animals].

PubMed

Segundo, Carolina; Martínez, Alejandrina; Arenas, Roberto; Fernández, Ramón; Cervantes, Roberto A

2004-03-01

Dermatophytic infections caused by M. canis in humans and animals have a world wide distribution and they are zoonotic. The objective in this work was to know the frequency of M. canis infections in humans and pets. We studied our cases from January 1994 to December 2002. The human samples were obtained from a Dermatological Department in a General Hospital and we registered the next data: age, sex, job, and affected area. The animal samples were obtained from a mycological veterinary laboratory, and we registered the presence or absence of clinical lesions. A total of 46 clinical cases of M. canis infections were recorded, 26 female and 20 males: tinea capitis 21, tinea corporis 17, tinea pedis five, onychomycosis two, and only one case with tinea faciei. The 46 cases with positive culture yield 42 positive samples in KOH. The age range varied from 2 to 60 years. Among the animals, we studied 461 dogs and found six KOH positive (1%) samples and cultured 23 isolates (4.98%): 21 M. canis, one M. gypseum and one Trichophyton spp. From the 68 samples of cats, eight (11.76%) were positive to KOH, being 26 (38.23%) M. canis isolates. In M. canis infections in humans, the age rage was wide with predominance in women. In animals, M. canis isolates represented the most dermatophytic infection.

Molecular Characterization of Shiga Toxin-Producing Escherichia coli Isolated from Ruminant and Donkey Raw Milk Samples and Traditional Dairy Products in Iran

PubMed Central

Momtaz, Hassan; Farzan, Rahil; Rahimi, Ebrahim; Safarpoor Dehkordi, Farhad; Souod, Negar

2012-01-01

The aims of the current study were to detect the virulence factors and antibiotic resistance of Shiga toxin-producing E. coli, in animal milk and dairy products in Iran. After E. coli dentification with culture method, PCR assay were developed for detection of pathogenic genes, serotypes and antibiotic resistance genes of E. coli. Results showed that out of 719 samples, 102 (14.18%) were confirmed to be positive for E. coli and out of 102 positive samples, 17.64% were O26 and 13.72% were O157 and 1.96% were O91 and 1.96% were O145 serotypes. Totally, the prevalence of stx1 and papA genes were the highest while the prevalence of sfaS and fyuA were the lowest in the positive samples. PCR results showed that tetA, tetB were the highest (64.70%) and aac(3)-IV were the lowest (27.45%) antibiotic resistant genes in E. coli positive samples. Our study indicated that the isolated E. coli trains in these regions had a highest antibiotic resistance to tetracycline (58.82%) and the lowest to nitrofurantoin (3.92%). tetA gene and E. coli O157 serotype had highest and aac(3)-IV gene, and E. coli O145 serotype had a lowest frequency rates of antibiotics resistance genes, in the region. PMID:22919299

Type-specific detection of high-risk human papillomavirus (HPV) in self-sampled cervicovaginal cells applied to FTA elute cartridge.

PubMed

Gustavsson, Inger; Sanner, Karin; Lindell, Monica; Strand, Anders; Olovsson, Matts; Wikström, Ingrid; Wilander, Erik; Gyllensten, Ulf

2011-08-01

Most procedures for self-sampling of cervical cells are based on liquid-based media for transportation and storage. An alternative is to use a solid support, such as dry filter paper media. To evaluate if self-sampling of cervicovaginal fluid using a cytobrush (Viba-brush; Rovers Medical Devices B.V., Oss, The Netherlands) and a solid support such as the Whatman Indicating FTA Elute cartridge (GE Healthcare, United Kingdom) can be used for reliable typing of human papillomavirus (HPV), as compared to cervical samples obtained by a physician using a cytobrush and the indicating FTA Elute Micro card and biopsy analysis. A total of 50 women with a previous high-risk (HR) HPV positive test were invited to perform self-sampling using the Viba-brush and the FTA cartridge and thereafter a physician obtained a cervical sample using the cytobrush and a FTA card, together with a cervical biopsy for histology and HPV typing. Detection of HR-HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59 was performed using three multiplex real-time polymerase chain reaction (PCR) assays. All samples contained sufficient amounts of genomic DNA and the self-samples yielded on average 3.5 times more DNA than those obtained by the physician. All women that were positive for HR-HPV in the biopsy sample also typed positive both by self-sampling and physician-obtained sampling. For women with a histological diagnosis of cervical intraepithelial neoplasia grades 2-3 (CIN 2-3) all three HPV samples showed 100% concordance. A higher number of women were HPV positive by self-sampling than by physician-obtained sampling or by biopsy analysis. The Viba-brush and the FTA cartridge are suitable for self-sampling of vaginal cells and subsequent HR-HPV typing. Copyright © 2011 Elsevier B.V. All rights reserved.

[Ophthalmopathy caused by precision work of sorters of precious stones].

PubMed

Feĭgin, A A; Korniushina, T A; Rozenblium, Iu Z

1992-01-01

A total of 440 female workers aged 17 to 50, whose work records ranged from 1 to 29 years, engaged in grading the diamonds by the color, shape, size, and quality (a total of 24 to 33 positions) were examined. A random sample of 110 subjects was singled out; this sample was divided into 2 equal groups with or without asthenopic complaints. The refraction, absolute accommodation volume, and relative accommodation reserves were under study. Comparison of these two groups of workers has shown that subjects with precision ophthalmopathy show a trend to a higher incidence of myopia, reduction of the absolute accommodation volume by 1.6 diopters and of the relative accommodation reserves by 1.3 diopters.

[Characteristics of lip-mouth region in smiling position from 80 adults with acceptable faces and individual normal occlusions].

PubMed

Zhang, J; Chen, Y; Zhou, X

1997-07-01

The Characteristics of Lip-mouth region including the soft and hard tissues in smiling position with frontal fixed position photographic computered analysis was studied. The examples were 80 adults (40 males and 40 females) with acceptable faces and individual normal occlusions. The age was between 17-25 years of age. The method adopting maximum smiling position to study the lip-mouth region is scientific, and the maximum smile line could be divided into three categories: low smile line-a-counted for 16.25% of the total samples; average smile line--68.75%; high smile line--15%.

Fast screening tests for the simultaneous detection of 11 drugs of abuse in urine specimens. A forensic epidemiology study of 28,298 cases in Tunisia.

PubMed

Moslah, B; Araoud, M; Nouioui, M A; Najjar, S; Amira, D; Ben Salah, N; Hedhili, A

2018-02-01

Forensic investigation performed on people suspected to be drug abusers covering all Tunisian cities was conducted by monitoring an epidemiological study of human urine samples surveying positive rates of consumption for drugs of abuse. The forensic investigations were conducted on a total of 28,298 arrested individuals suspected to be drug addicts during five years (January 2010-December 2015). An immunoassay screening tests to detect elevated levels of drugs classes in urine samples was performed. These screening assays provide a preliminary qualitative test result. Only positives urine specimens were analyzed with GC-MS for confirmation. Except for cannabis, the results showed insignificant number of positive cases for cocaine, ecstasy (MDMA) and amphetamine consumptions (<1%). Copyright © 2017 Elsevier B.V. All rights reserved.

Isolation and molecular characterization of Vibrio parahaemolyticus from fresh, low-temperature preserved, dried, and salted seafood products in two coastal areas of eastern China.

PubMed

Yang, Zhen-Quan; Jiao, Xin-An; Zhou, Xiao-Hui; Cao, Guo-Xiang; Fang, Wei-Ming; Gu, Rui-Xia

2008-07-31

A total of 1293 seafood samples from fishing farm, retail markets, restaurants and cooking rooms of hotels in Jiangsu province and Shanghai city of China were collected and analyzed for the prevalence of Vibrio parahaemolyticus during July to October in 2007. Two hundred and fifty one isolates of V. parahaemolyticus were identified, of which 8 isolates were positive for tdh and 2 were positive for trh gene. Three tdh positive isolates were identified from low-temperature preserved seafood samples and 5 isolates from fresh seafood samples, of these tdh positive isolates, 3 were positive in ORF8-PCR test. The genetic diversity among V. parahaemolyticus isolates was assessed using random amplified polymorphic DNA (RAPD)-PCR and the results showed that there were 33 different genetic patterns that were clustered into nine groups (groups A to I) at 82% similarity level. About 31.9% of the isolates belong to type III9d that were widely distributed in fresh, iced, frozen, dried and salted seafood samples. Seven tdh positive isolates belonged to group A and one belonged to group C, 2 trh positive isolates were type I10d belonging to group F, which was identical to that of reference strains isolated from patients. This study demonstrated genetic variability within V. parahaemolyticus isolates from seafood in Chinese markets and confirmed the presence of toxigenic V. parahaemolyticus not only in fresh but also in iced and frozen seafood products indicating that low-temperature preserved seafood might be also a vehicle for transmitting pathogenic V. parahaemolyticus.

Correlation between sperm DNA fragmentation index and CMA3 positive spermatozoa in globozoospermic patients.

PubMed

Hosseinifar, H; Yazdanikhah, S; Modarresi, T; Totonchi, M; Sadighi Gilani, M A; Sabbaghian, M

2015-05-01

The absence of the acrosome causes the situation which is called globozoospermia. There are a few studies, mostly as case reports, about correlation between levels of sperm DNA damage in patients with total round-headed spermatozoa. We investigated this correlation as well as CMA3 positive spermatozoa in 20 globozoospermic men (with more than 90% round-headed spermatozoa) attending to Royan Institute. Semen samples divided into three parts to semen analysis, to measure DNA fragmentation index (DFI) using sperm chromatin structure assay (SCSA) and to detect CMA3(+) sperm cells by chromomycin A3 staining and fluorescent microscopy. Our results showed that there were significant differences in sperm concentration, total sperm motility, and normal morphology between patients and controls group (p < 0.001). Moreover, the average of DFI and CMA3 positive spermatozoa in patients group significantly increases compared with control group (p < 0.001). A significant correlation between DFI and CMA3(+) in total population was also detected in patients group (r = 0.45, p = 0.046). To our knowledge, this is the largest study about correlation between DNA damage levels and CMA3 positive spermatozoa with round head sperm cells in total globozoospermic men. It seems that the increase in DNA damage may be because of defective sperm DNA compaction, as we detected CMA3 positive sperm cells in these patients. © 2015 American Society of Andrology and European Academy of Andrology.

Auto-calibrated scanning-angle prism-type total internal reflection microscopy for nanometer-precision axial position determination and optional variable-illumination-depth pseudo total internal reflection microscopy

DOEpatents

Fang, Ning; Sun, Wei

2015-04-21

A method, apparatus, and system for improved VA-TIRFM microscopy. The method comprises automatically controlled calibration of one or more laser sources by precise control of presentation of each laser relative a sample for small incremental changes of incident angle over a range of critical TIR angles. The calibration then allows precise scanning of the sample for any of those calibrated angles for higher and more accurate resolution, and better reconstruction of the scans for super resolution reconstruction of the sample. Optionally the system can be controlled for incident angles of the excitation laser at sub-critical angles for pseudo TIRFM. Optionally both above-critical angle and sub critical angle measurements can be accomplished with the same system.

Comparing microbial water quality in an intermittent and continuous piped water supply.

PubMed

Kumpel, Emily; Nelson, Kara L

2013-09-15

Supplying piped water intermittently is a common practice throughout the world that increases the risk of microbial contamination through multiple mechanisms. Converting an intermittent supply to a continuous supply has the potential to improve the quality of water delivered to consumers. To understand the effects of this upgrade on water quality, we tested samples from reservoirs, consumer taps, and drinking water provided by households (e.g. from storage containers) from an intermittent and continuous supply in Hubli-Dharwad, India, over one year. Water samples were tested for total coliform, Escherichia coli, turbidity, free chlorine, and combined chlorine. While water quality was similar at service reservoirs supplying the continuous and intermittent sections of the network, indicator bacteria were detected more frequently and at higher concentrations in samples from taps supplied intermittently compared to those supplied continuously (p < 0.01). Detection of E. coli was rare in continuous supply, with 0.7% of tap samples positive compared to 31.7% of intermittent water supply tap samples positive for E. coli. In samples from both continuously and intermittently supplied taps, higher concentrations of total coliform were measured after rainfall events. While source water quality declined slightly during the rainy season, only tap water from intermittent supply had significantly more indicator bacteria throughout the rainy season compared to the dry season. Drinking water samples provided by households in both continuous and intermittent supplies had higher concentrations of indicator bacteria than samples collected directly from taps. Most households with continuous supply continued to store water for drinking, resulting in re-contamination, which may reduce the benefits to water quality of converting to continuous supply. Copyright © 2013 Elsevier Ltd. All rights reserved.

Detection of Total and Pathogenic Vibrio parahaemolyticus in Shellfish Growing along the South Yellow Sea and the East China Sea.

PubMed

Han, Feng; Gu, Run-Run; Shen, Xiao-Sheng; Chen, Yuan-Ge; Tian, Liang-Liang; Zhou, Wei-Feng; Cai, You-Qiong

2017-10-17

This study was conducted to monitor the densities of total and pathogenic Vibrio parahaemolyticus in 300 samples of nine shellfish species harvested from the coasts of the South Yellow Sea and the East China Sea (N 23° to 34°, E 116° to 124°), People's Republic of China, between May and October 2015. Total V. parahaemolyticus densities were measured, and V. parahaemolyticus isolates were biochemically identified with probes for the thermostable direct hemolysin gene (tdh) and the thermostable direct hemolysin-related hemolysin gene (trh). We found that 202 of the 300 samples were positive for V. parahaemolyticus from all the sites: 58 of the 100 samples from the Fujian province, 71 of the 100 samples from the Zhejiang province, and 73 of the 100 samples from the Jiangsu province. In most (170) of the 300 samples, V. parahaemolyticus densities were 0.3 to 10 most probable number (MPN)/g; five lots exceeded 110 MPN/g, and two lots were estimated at 110 MPN/g. Among the 202 V. parahaemolyticus strains, only one was trh positive. Densities of V. parahaemolyticus in these shellfish were temperature dependent, with highest densities in June and July. Among the nine mollusk species, V. parahaemolyticus was most abundant in the agemaki clam (Sinonovacula constricta). The highest and lowest V. parahaemolyticus prevalences were found in oriental cyclina (Cyclina sinensis, 93.8%) and mussels (Mytilus edulis, 28.1%), respectively. Overall, although V. parahaemolyticus is widely distributed in marine environments, the density of V. parahaemolyticus was low and the prevalence of the main virulence factor was very low in shellfish along the coasts of the South Yellow Sea and East China Sea, which is important from a public health perspective. Data presented here will be useful for correlational research and can be utilized for developing risk management plans that establish food safety guidelines for V. parahaemolyticus in Chinese shellfish.

Mutagenicity of drinking water sampled from the Yangtze River and Hanshui River (Wuhan section) and correlations with water quality parameters.

PubMed

Lv, Xuemin; Lu, Yi; Yang, Xiaoming; Dong, Xiaorong; Ma, Kunpeng; Xiao, Sanhua; Wang, Yazhou; Tang, Fei

2015-03-31

A total of 54 water samples were collected during three different hydrologic periods (level period, wet period, and dry period) from Plant A and Plant B (a source for Yangtze River and Hanshui River water, respectively), and several water parameters, such as chemical oxygen demand (COD), turbidity, and total organic carbon (TOC), were simultaneously analyzed. The mutagenicity of the water samples was evaluated using the Ames test with Salmonella typhimurium strains TA98 and TA100. According to the results, the organic compounds in the water were largely frame-shift mutagens, as positive results were found for most of the tests using TA98. All of the finished water samples exhibited stronger mutagenicity than the relative raw and distribution water samples, with water samples collected from Plant B presenting stronger mutagenic strength than those from Plant A. The finished water samples from Plant A displayed a seasonal-dependent variation. Water parameters including COD (r = 0.599, P = 0.009), TOC (r = 0.681, P = 0.02), UV254 (r = 0.711, P = 0.001), and total nitrogen (r = 0.570, P = 0.014) exhibited good correlations with mutagenicity (TA98), at 2.0 L/plate, which bolsters the argument of the importance of using mutagenicity as a new parameter to assess the quality of drinking water.

Mutagenicity of drinking water sampled from the Yangtze River and Hanshui River (Wuhan section) and correlations with water quality parameters

PubMed Central

Lv, Xuemin; Lu, Yi; Yang, Xiaoming; Dong, Xiaorong; Ma, Kunpeng; Xiao, Sanhua; Wang, Yazhou; Tang, Fei

2015-01-01

A total of 54 water samples were collected during three different hydrologic periods (level period, wet period, and dry period) from Plant A and Plant B (a source for Yangtze River and Hanshui River water, respectively), and several water parameters, such as chemical oxygen demand (COD), turbidity, and total organic carbon (TOC), were simultaneously analyzed. The mutagenicity of the water samples was evaluated using the Ames test with Salmonella typhimurium strains TA98 and TA100. According to the results, the organic compounds in the water were largely frame-shift mutagens, as positive results were found for most of the tests using TA98. All of the finished water samples exhibited stronger mutagenicity than the relative raw and distribution water samples, with water samples collected from Plant B presenting stronger mutagenic strength than those from Plant A. The finished water samples from Plant A displayed a seasonal-dependent variation. Water parameters including COD (r = 0.599, P = 0.009), TOC (r = 0.681, P = 0.02), UV254 (r = 0.711, P = 0.001), and total nitrogen (r = 0.570, P = 0.014) exhibited good correlations with mutagenicity (TA98), at 2.0 L/plate, which bolsters the argument of the importance of using mutagenicity as a new parameter to assess the quality of drinking water. PMID:25825837

Simplicimonas-like DNA in vaginal swabs of cows and heifers cross-reacting in the real-time PCR for T. foetus.

PubMed

Frey, Caroline F; Müller, Norbert; Stäuber, Norbert; Marreros, Nelson; Hofmann, Larissa; Hentrich, Brigitte; Hirsbrunner, Gaby

2017-04-15

Cows on an alpine pasture were presented with severe signs of vaginitis. To rule out infection with Tritrichomonas foetus, vaginal swabs were taken and real-time PCR based on detection via fluorescence resonance energy transfer (FRET) probes and targeting the first internal transcribed spacer (ITS-1) of nuclear ribosomal DNA (rDNA) was performed. PCR was positive in 25 of totally 34 assessed cows. However, the melting profiles of the probes targeting the diagnostic PCR products differed from the T. foetus positive control. Subsequent sequencing of the amplicons revealed 91% identity to Simplicimonas sp. sequences deposited in GenBank™. Furthermore, there was no clear association between positive PCR result and presence of vaginitis. To investigate the distribution of this Simplicimonas-like organism in cows, more herds grazing on the same alpine pastures as well as unrelated cows were tested. In total, 133 cows and 16 heifers were sampled, 53 cows and 6 heifers even twice. Vaginitis was evident in 43 cows and 4 heifers. All-over-positivity of PCR was 44%, including nine tests performed on heifers. Melting peak analysis indicated Simplicimonas-like organisms in all positive samples. Culture attempts in bovine InPouch ™ TF failed. No association between a positive PCR result and the presence of vaginitis was found. This is, to the best of our knowledge, the first report on Simplicimonas-like DNA in vaginal swabs of female cattle. Our data suggest that when testing vaginal swabs of cattle by means of T. foetus PCR, false positive reactions due to Simplicimonas-like organisms may occur. Copyright © 2017 Elsevier B.V. All rights reserved.

Feature genes predicting the FLT3/ITD mutation in acute myeloid leukemia

PubMed Central

LI, CHENGLONG; ZHU, BIAO; CHEN, JIAO; HUANG, XIAOBING

2016-01-01

In the present study, gene expression profiles of acute myeloid leukemia (AML) samples were analyzed to identify feature genes with the capacity to predict the mutation status of FLT3/ITD. Two machine learning models, namely the support vector machine (SVM) and random forest (RF) methods, were used for classification. Four datasets were downloaded from the European Bioinformatics Institute, two of which (containing 371 samples, including 281 FLT3/ITD mutation-negative and 90 mutation-positive samples) were randomly defined as the training group, while the other two datasets (containing 488 samples, including 350 FLT3/ITD mutation-negative and 138 mutation-positive samples) were defined as the test group. Differentially expressed genes (DEGs) were identified by significance analysis of the micro-array data by using the training samples. The classification efficiency of the SCM and RF methods was evaluated using the following parameters: Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and the area under the receiver operating characteristic curve. Functional enrichment analysis was performed for the feature genes with DAVID. A total of 585 DEGs were identified in the training group, of which 580 were upregulated and five were downregulated. The classification accuracy rates of the two methods for the training group, the test group and the combined group using the 585 feature genes were >90%. For the SVM and RF methods, the rates of correct determination, specificity and PPV were >90%, while the sensitivity and NPV were >80%. The SVM method produced a slightly better classification effect than the RF method. A total of 13 biological pathways were overrepresented by the feature genes, mainly involving energy metabolism, chromatin organization and translation. The feature genes identified in the present study may be used to predict the mutation status of FLT3/ITD in patients with AML. PMID:27177049

Substantial variation in the hepatitis B surface antigen (HBsAg) in hepatitis B virus (HBV)-positive patients from South Africa: Reliable detection of HBV by the Elecsys HBsAg II assay.

PubMed

Gencay, Mikael; Vermeulen, Marion; Neofytos, Dionysis; Westergaard, Gaston; Pabinger, Stephan; Kriegner, Albert; Seffner, Anja; Gohl, Peter; Huebner, Kirsten; Nauck, Markus; Kaminski, Wolfgang E

2018-04-01

It is essential that hepatitis B surface antigen (HBsAg) diagnostic assays reliably detect genetic diversity in the major hydrophilic region (MHR) of HBsAg to avoid false-negative results. Mutations in this domain display marked ethno-geographic variation and may lead to failure to diagnose hepatitis B virus (HBV) infection. Evaluate diagnostic performance of the Elecsys ® HBsAg II Qualitative assay in a cohort of South African HBV-positive blood donors. A total of 179 South African HBsAg- and HBV DNA > 100 IU/mL-positive blood donor samples were included. Samples were sequenced for genetic variation in HBsAg MHR using next-generation ultra-deep sequencing. HBsAg seropositivity was determined using the Roche Elecsys HBsAg II Qualitative assay. Mutation rates were compared between the first (amino acids 124-137) and second (amino acids 139-147) loops of the immunodominant MHR 'a' determinant region. Frequency of occult HBV infection-associated Y100C mutations was also determined. We observed a total of 279 MHR mutations (117 variants) in 102 (57%) samples, of which 91 were located in the 'a' determinant region. The major vaccine-induced escape mutation G145R was observed in two samples. All occult HBV infection-associated Y100C and common diagnostic and vaccine-escape-associated P120T, G145R, K122R, M133L, M133T, Q129H, G130N, and T126S mutations were reliably detected by the assay, which consistently detected the presence of HBsAg in all 179 samples including samples with 11 novel mutations. Despite substantial variation in HBsAg MHR, the Elecsys HBsAg II Qualitative assay robustly detects HBV infection in this South African cohort. Copyright © 2018 The Author(s). Published by Elsevier B.V. All rights reserved.

Feature genes predicting the FLT3/ITD mutation in acute myeloid leukemia.

PubMed

Li, Chenglong; Zhu, Biao; Chen, Jiao; Huang, Xiaobing

2016-07-01

In the present study, gene expression profiles of acute myeloid leukemia (AML) samples were analyzed to identify feature genes with the capacity to predict the mutation status of FLT3/ITD. Two machine learning models, namely the support vector machine (SVM) and random forest (RF) methods, were used for classification. Four datasets were downloaded from the European Bioinformatics Institute, two of which (containing 371 samples, including 281 FLT3/ITD mutation-negative and 90 mutation‑positive samples) were randomly defined as the training group, while the other two datasets (containing 488 samples, including 350 FLT3/ITD mutation-negative and 138 mutation-positive samples) were defined as the test group. Differentially expressed genes (DEGs) were identified by significance analysis of the microarray data by using the training samples. The classification efficiency of the SCM and RF methods was evaluated using the following parameters: Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and the area under the receiver operating characteristic curve. Functional enrichment analysis was performed for the feature genes with DAVID. A total of 585 DEGs were identified in the training group, of which 580 were upregulated and five were downregulated. The classification accuracy rates of the two methods for the training group, the test group and the combined group using the 585 feature genes were >90%. For the SVM and RF methods, the rates of correct determination, specificity and PPV were >90%, while the sensitivity and NPV were >80%. The SVM method produced a slightly better classification effect than the RF method. A total of 13 biological pathways were overrepresented by the feature genes, mainly involving energy metabolism, chromatin organization and translation. The feature genes identified in the present study may be used to predict the mutation status of FLT3/ITD in patients with AML.

Clinical Follow-Up of Responses to Treatment with Benznidazol in Amazon: A Cohort Study of Acute Chagas Disease

PubMed Central

Pinto, Ana Yecê das Neves; Valente, Vera da Costa; Coura, José Rodrigues; Valente, Sebastião Aldo da Silva; Junqueira, Angela Cristina Veríssimo; Santos, Laura Cristina; Ferreira, Alberto Gomes; de Macedo, Roberto Cavalleiro

2013-01-01

A total of 179 individuals with acute Chagas disease mainly transmitted by oral source, from Pará and Amapá State, Amazonian, Brazil were included during the period from 1988 to 2005. Blood samples were used to survey peripheral blood for T. cruzi hemoparasites by quantitative buffy coat (QBC), indirect xenodiagnosis, blood culture and serology to detection of total IgM and anti-T. cruzi IgG antibodies by indirect immunofluorescence assay (IFA) and indirect hemagglutination assay (HA). All assays were performed pre-treatment (0 days) and repeated 35 (±7) and 68 (±6) days after the initiation of treatment with benznidazol and every 6 months while remained seropositive. The endpoint of collection was performed in 2005. Total medium period of follow-up per person was 5.6 years. Also, a blood sample was collected from 72 randomly chosen treated patients to perform polimerase chain reaction (PCR) method. Proportions of subjects with negative or positive serology according to the number of years after treatment were compared. In the endpoint of follow-up we found 47 patients (26.7%) serologically negative, therefore considered cured and 5 (2.7%) exhibited mild cardiac Chagas disease. Other 132 patients had persistent positive serologic tests. The PCR carried out in 72 individuals was positive in 9.8%. Added, there was evidence of therapeutic failure immediately following treatment, as demonstrated by xenodiagnosis and blood culture methods in 2.3% and 3.5% of cases, respectively. There was a strong evidence of antibody clearing in the fourth year after treatment and continuous decrease of antibody titers. Authors suggest that control programs should apply operational researches with new drug interventions four years after the acute phase for those treated patients with persistently positive serology. PMID:23724050

Longitudinal study of Escherichia coli O157 shedding and super shedding in dairy heifers.

PubMed

Williams, K J; Ward, M P; Dhungyel, O P

2015-04-01

A longitudinal study was conducted to assess the methods available for detection of Escherichia coli O157 and to investigate the prevalence and occurrence of long-term shedding and super shedding in a cohort of Australian dairy heifers. Samples were obtained at approximately weekly intervals from heifers at pasture under normal management systems. Selective sampling techniques were used with the aim of identifying heifers with a higher probability of shedding or super shedding. Rectoanal mucosal swabs (RAMS) and fecal samples were obtained from each heifer. Direct culture of feces was used for detection and enumeration. Feces and RAMS were tested by enrichment culture. Selected samples were further tested retrospectively by immunomagnetic separation of enriched samples. Of 784 samples obtained, 154 (19.6%) were detected as positive using culture methods. Adjusting for selective sampling, the prevalence was 71 (15.6%) of 454. In total, 66 samples were detected as positive at >10(2) CFU/g of which 8 were >10(4) CFU/g and classed as super shedding. A significant difference was observed in detection by enriched culture of RAMS and feces. Dairy heifers within this cohort exhibited variable E. coli O157 shedding, consistent with previous estimates of shedding. Super shedding was detected at a low frequency and inconsistently from individual heifers. All detection methods identified some samples as positive that were not detected by any other method, indicating that the testing methods used will influence survey results.

[A comparison of soil contamination with Toxocara canis and Toxocara cati eggs in rural and urban areas of Wielkopolska district in 2000-2005].

PubMed

Mizgajska-Wiktor, Hanna; Jarosz, Wojciech

2007-01-01

The aim of the studies was to compare the degree of soil contamination with Toxocara canis and T. cati eggs in rural and urban areas depending on time of sampling and type of places examined. Material and methods. Over 2000-2005 a total of 538 soil samples from 3 villages and 368 from Poznań city (Poland) areas were examined for Toxocara spp. eggs. In spring 418 samples in rural areas and 184 samples in urban areas were collected and in autumn 120 and 184 respectively. The samples were examined using flotation technique in saturated sodium nitrate. The discrimination of T. canis and T. cati eggs was based on the size of eggs and transparency of shell layers. Results. The contamination of soil with Toxocara eggs was higher in the urban areas (19.8% positive samples) than in the rural ones (15.6% positive samples) and city or village-backyards were most heavily contaminated. Both, in the villages and in the city, the degree of soil contamination with eggs in spring and autumn was similar (17.6 and 14.8% positive samples respectively). T. cati eggs were much more prevalent in urban areas (97% of all eggs recovered) while T. canis in rural areas (84% of all recovered eggs). The share of T. canis and T. cati eggs in soil contamination did not depend on the time of sampling.

Clinicopathological Implications of Human Papilloma Virus (HPV) L1 Capsid Protein Immunoreactivity in HPV16-Positive Cervical Cytology

PubMed Central

Lee, Sung-Jong; Lee, Ah-Won; Kang, Chang-Suk; Park, Jong-Sup; Park, Dong-Choon; Ki, Eun-Young; Lee, Keun-Ho; Yoon, Joo-Hee; Hur, Soo-Young; Kim, Tae-Jung

2014-01-01

Background: The objective of this study was to investigate the expression of human papilloma virus (HPV) L1 capsid protein in abnormal cervical cytology with HPV16 infection and analyze its association with cervical histopathology in Korean women. Material and Methods: We performed immunocytochemistry for HPV L1 in 475 abnormal cervical cytology samples from patients with HPV16 infections using the Cytoactiv® HPV L1 screening set. We investigated the expression of HPV L1 in cervical cytology samples and compared it with the results of histopathological examination of surgical specimens. Results: Of a total of 475 cases, 188 (39.6%) were immunocytochemically positive and 287 (60.4%) negative for HPV L1. The immunocytochemical expression rates of HPV L1 in atypical squamous cells of unknown significance (ASCUS), low-grade squamous intraepithelial lesions (LSIL), high-grade squamous intraepithelial lesions (HSIL), and cancer were 21.8%, 59.7%, 19.1%, and 0.0%, respectively. LSIL exhibited the highest rate of HPV L1 positivity. Of a total of 475 cases, the multiple-type HPV infection rate, including HPV16, in HPV L1-negative cytology samples was 27.5%, which was significantly higher than that in HPV L1-positive cytology samples (p = 0.037). The absence of HPV L1 expression in ASCUS and LSIL was significantly associated with high-grade (≥cervical intraepithelial neoplasia [CIN] 2) than low-grade (≤CIN1) histopathology diagnoses (p < 0.05), but was not significantly different between HPV16 single and multiple-type HPV infections (p > 0.05). On the other hand, among 188 HPV L1-positive cases, 30.6% of multiple-type HPV infections showed high-grade histopathology diagnoses (≥CIN3), significantly higher than the percentage of HPV16 single infections (8.6%) (p = 0.0004) Conclusions: Our study demonstrates that the expression of HPV L1 is low in advanced dysplasia. Furthermore, the absence of HPV L1 in HPV16-positive low-grade cytology (i.e., ASCUS and LSIL) is strongly associated with high-grade histopathology diagnoses. The multiplicity of HPV infections may have an important role in high-grade histopathology diagnoses (≥CIN3) in HPV L1-positive cases. PMID:24396289

Long-Term Memory for Knowledge Learned in School.

ERIC Educational Resources Information Center

Semb, George B.; And Others

1993-01-01

Three experiments (initial total sample of 190 college students) examined students' long-term retention of knowledge learned in college courses. Results of all experiments suggest that students remember a great deal of what they learn up to 11 months later and that tutoring has positive effects on recall by tutors. (SLD)

Habitat use and trophic position effects on contaminant bioaccumulation in St. Louis River Estuary fishes

EPA Science Inventory

The objective of our study was to determine the relationship between fish tissue stable isotope composition and total mercury or polychlorinated biphenyl (PCB) concentrations in the St. Louis River estuary food web. We sampled two resident fishes, Yellow Perch (Perca flavescens) ...

Habitat use and trophic position effects on contaminant bioaccumulation in fish indicated by stable isotope composition

EPA Science Inventory

The objective of our study was to determine the relationship between fish tissue stable isotope composition and total mercury or polychlorinated biphenyl (PCB) concentrations in a Great Lakes coastal food web. We sampled two resident fishes, Yellow Perch (Perca flavescens) and Bl...

Diversity of cuticular wax among Salix species and Populus species hybrids.

PubMed

Cameron, Kimberly D; Teece, Mark A; Bevilacqua, Eddie; Smart, Lawrence B

2002-08-01

The leaf cuticular waxes of three Salix species and two Populus species hybrids, selected for their ability to produce high amounts of biomass, were characterized. Samples were extracted in CH(2)Cl(2) three times over the growing season. Low kV SEM was utilized to observe differences in the ultrastructure of leaf surfaces from each clone. Homologous series of wax components were classified into organic groups, and the variation in wax components due to clone, sample time, and their interaction was identified. All Salix species and Populus species hybrids showed differences in total wax load at each sampling period, whereas the pattern of wax deposition over time differed only between the Salix species. A strong positive relationship was identified between the entire homologous series of alcohols and total wax load in all clones. Similarly strong relationships were observed between fatty acids and total wax load as well as fatty acids and alcohols in two Salix species and one Populus species hybrid. One Salix species, S. dasyclados, also displayed a strong positive relationship between alcohols and alkanes. These data indicate that species grown under the same environmental conditions produce measurably different cuticular waxes and that regulation of wax production appears to be different in each species. The important roles cuticular waxes play in drought tolerance, pest, and pathogen resistance, as well as the ease of wax extraction and analysis, strongly suggest that the characteristics of the cuticular wax may prove to be useful selectable traits in a breeding program.

Genus Level Identification of Mycobacteria from Clinical Specimens by Using an Easy-To-Handle Mycobacterium-Specific PCR Assay

PubMed Central

Stauffer, Fritz; Haber, Heinrich; Rieger, Armin; Mutschlechner, Robert; Hasenberger, Petra; Tevere, Vincent J.; Young, Karen K. Y.

1998-01-01

An easy-to-handle Mycobacterium-specific PCR assay for detection of the presence of a wide range of mycobacterial species in clinical samples was evaluated. The performance of the genus probe was compared with the performance of probes specific for Mycobacterium tuberculosis and Mycobacterium avium and with that of standard culture. In addition, the utility of an internal control in monitoring amplification inhibitors was studied. Of 545 respiratory and 325 nonrespiratory specimens (a total of 870 specimens), 58 (6.7%) showed the presence of amplification inhibitors, as determined by a negative result for the internal control. Of these 58 specimens, 31 (53%) were stool specimens; other material, even citrate blood after lysis of erythrocytes, did not pose a problem with regard to inhibition of PCR amplification. Eighty-one of the remaining 812 specimens had a positive Mycobacterium culture result. Of these culture-positive specimens, 58 (71.6%) showed a positive result with the Mycobacterium genus-specific probe. Seventy-two samples had a positive result with the Mycobacterium-specific probe but a negative culture result. Of these 72 samples, 26 samples were regarded as true positive, either because the M. tuberculosis- or M. avium-specific probe was also positive at the same time or because other specimens from the same patient taken at the same time were culture positive. The sensitivity of the Mycobacterium-specific probe was 78.5% and the specificity was 93.5%. This study showed that pretesting of clinical specimens for mycobacteria to the genus level with a Mycobacterium-specific probe offers the routine clinical laboratory the possibility of detecting tuberculous and nontuberculous mycobacteria with one test. Furthermore, specimens testing positive with the genus-specific probe can be immediately identified with species-specific probes. PMID:9508282

Rapid detection of coliforms in drinking water of Arak city using multiplex PCR method in comparison with the standard method of culture (Most Probably Number)

PubMed Central

Fatemeh, Dehghan; Reza, Zolfaghari Mohammad; Mohammad, Arjomandzadegan; Salomeh, Kalantari; Reza, Ahmari Gholam; Hossein, Sarmadian; Maryam, Sadrnia; Azam, Ahmadi; Mana, Shojapoor; Negin, Najarian; Reza, Kasravi Alii; Saeed, Falahat

2014-01-01

Objective To analyse molecular detection of coliforms and shorten the time of PCR. Methods Rapid detection of coliforms by amplification of lacZ and uidA genes in a multiplex PCR reaction was designed and performed in comparison with most probably number (MPN) method for 16 artificial and 101 field samples. The molecular method was also conducted on isolated coliforms from positive MPN samples; standard sample for verification of microbial method certificated reference material; isolated strains from certificated reference material and standard bacteria. The PCR and electrophoresis parameters were changed for reducing the operation time. Results Results of PCR for lacZ and uidA genes were similar in all of standard, operational and artificial samples and showed the 876 bp and 147 bp bands of lacZ and uidA genes by multiplex PCR. PCR results were confirmed by MPN culture method by sensitivity 86% (95% CI: 0.71-0.93). Also the total execution time, with a successful change of factors, was reduced to less than two and a half hour. Conclusions Multiplex PCR method with shortened operation time was used for the simultaneous detection of total coliforms and Escherichia coli in distribution system of Arak city. It's recommended to be used at least as an initial screening test, and then the positive samples could be randomly tested by MPN. PMID:25182727

Do elevated nutrients and organic carbon on Philippine reefs increase the prevalence of coral disease?

NASA Astrophysics Data System (ADS)

Kaczmarsky, L.; Richardson, L. L.

2011-03-01

Characterizations of Philippine coral diseases are very limited. The two most common, ulcerative white spot disease (UWS) and massive Porites growth anomalies (MPGA), target the genus Porites, a dominant reef-building genus. This is the first investigation in the Philippines to detect positive correlations between coral disease, nutrient levels, and organic carbon. A total of 5,843 Porites colonies were examined. Water and sediment samples were collected for analyses of nutrients (total nitrogen and phosphorus) and total organic carbon at 15 sites along a 40.5 km disease gradient, which was previously shown to positively correlate with human population levels. Results suggest that outbreaks of UWS and MPGAs are driven by elevated nutrient and organic carbon levels. Although the variables analyzed could be proxies for other causative agents (e.g., high sediment levels), the results provide quantitative evidence linking relatively higher coral disease prevalence to an anthropogenically impacted environment.

Microbiological Survey of Packaged Ready-to-Eat Red Meats at Retail in New Zealand.

PubMed

Rivas, Lucia; Horn, Beverley; Cook, Roger; Castle, Marion

2017-10-04

A microbiological survey was undertaken on packaged ready-to-eat red meats available at retail in New Zealand. A total of 1,485 samples (297 lots of five samples each) were collected according to a sampling plan based on market share and regulatory regimes (Animal Products Act 1999 and Food Act 1981) and were tested against the microbiological limits specified in Food Standards Code (FSC) 1.6.1 applicable at the time of sampling. Each lot was tested as a composite for the presence or absence of Salmonella spp., coagulase-producing staphylococci, Listeria monocytogenes, and other Listeria spp. at the end of the manufacturer's stated shelf life. Individual samples within a positive lot were subsequently enumerated for L. monocytogenes. None of the samples contained Salmonella spp. or had coagulase-producing staphylococci counts above the acceptable level specified in FSC 1.6.1 (>100 CFU/g). Data showed that 93.6% (278 of 297 lots) of ready-to-eat red meat complied with the FSC 1.6.1 criteria applicable at the time of the survey. The failure of 19 lots (6.4%) was due to the presence of L. monocytogenes from product obtained from 8 of 33 producers tested. Thirteen samples of 95 positive samples were found to contain between 50 and 500 CFU/g L. monocytogenes, but all of these samples were manufactured by the same operator. Pulsed-field gel electrophoresis typing of all of the L. monocytogenes isolates obtained from the survey identified 12 different pulsotypes. Different pulsotypes were often identified in samples from the same operator sampled on separate occasions. A total of 46 lots (15.5%) contained Listeria spp. (including L. monocytogenes). The detection of Listeria in samples may highlight the existence of problems in operator processing and/or packaging processes and suggests that improvements in good hygienic practice and implementation of more effective risk mitigation strategies are needed.

[Evaluation of Mascia Brunelli rapid antigen test in the diagnosis of group A streptococcal pharyngitis].

PubMed

Barış, Ayşe; Anlıaçık, Nur; Bulut, Mehmet Emin; Deniz, Rıdvan; Yücel, Elif; Aktaş, Elif

2017-01-01

Pharyngitis in most cases is due to viral microorganisms however drug therapy without the detection of etiological agent leads to unnecessary use of antibiotics. On the other hand, when the etiologic agent is group A beta-hemolytic streptococci (GAS) it is important to identify the etiologic agent rapidly which will guide the treatment with appropriate antibiotics. The use of highly sensitive rapid tests will contribute significantly to early diagnosis and appropriate therapy. The aim of this study is to evaluate the efficacy of Mascia Brunelli rapid antigen test for the detection of GAS in throat swab samples. A total of 833 throat swab samples submitted to our laboratory with pre-diagnosis of pharyngitis were assessed between June 2016 and August 2016. The samples were simultaneously cultured and tested by rapid Mascia Brunelli Strep-A Card (Mascia Brunelli S.p.a, Italy). For identification, bacitracin sensitivity, PYR test and latex agglutination test in addition to Bruker MALDI-TOF MS (Daltonics, Germany) system were used. The density of GAS growth in the culture was noted. The samples that were false negative with Mascia Brunelli test were re-tested with QuickVue + Strep A Test (Quidel Corporation, San Diego, USA) rapid antigen test. A total of 833 patients, 376 (45.2%) female and 457 (54.8%) male were included in the study. The age range was between 0-94 years with a mean value of 7.86 ± 6.72. 125 (15%) and 94 (11.28%) of the samples were positive with culture and rapid antigen test, respectively. Mascia Brunelli antigen test gave negative results for 31 culture positive samples. Of these 31 samples, 28 were found positive by QuickVue + Strep A antigen test. As a result, the sensitivity of the test was found to be independent of the inoculum effect. The culture positivity rate in patients between 5-15 years was 18.4%. The sensitivity, specificity, positive predictive value, negative predictive value and the accuracy of Mascia Brunelli antigen test, with respect to culture, were 75.2%, 100%, 100%, 95.81% and 96.28%, respectively. In conclusion, the selection of rapid antigen tests with high sensitivity in the diagnosis of GAS pharyngitis will contribute to the prevention of resistance development by appropriate use of antibiotics as well as early diagnosis and appropriate treatment. However, confirmation of negative rapid antigen test results by culture is very important in terms of false diagnosis and prevention of incomplete treatment.

Molecular identification of Blastocystis sp. subtypes in water samples collected from Black sea, Turkey.

PubMed

Koloren, Zeynep; Gulabi, Berivan Basak; Karanis, Panagiotis

2018-04-01

The aim of this study was to identify the subtypes of Blastocystis sp. and complete a phylogenetic analysis of 268 water samples that were collected from the Samsun, Amasya and Sinop Provinces of the Black Sea in Turkey, between the years 2011 and 2014. Blastocystis sp. was investigated in 48 uncultured sea water samples that were collected from 4 sites within the Sinop Province. A total of 100 river water samples were collected from 37 sites in the Samsun Province and 120 river water samples were collected from 10 sampling sites within the Amasya Province. The small subunit (SSU) rDNA gene Polymerase chain reaction (PCR) were performed for the detection of Blastocytis sp. and the PCR-positive samples were sequenced. Subsequently, the (SSU) rDNA sequences were aligned by Bioedit and phylogenetic trees were constructed for Blastocystis with reference to the genotypes from GenBank. Blastocystis sp. were found in 3 out of the 75 (4%) river water samples that were collected from the Samsun Province. Six of the 120 (5%) river water samples and 1 out of the 48 (2%) seawater samples were positive for Blastocystis in the Amasya and Sinop Provinces. There were two different subtypes (ST; 1 and 3) found from sequencing all of the samples from the investigated sites. Two and one PCR products were found to be positive for ST1 and ST3 from the different samples collected within the Samsun Province. Two and 4 PCR products from the Amasya Province were ST1 and ST3, respectively and only one sample from the Sinop Province was found to be positive for ST1. This is the first report to identify and report the occurrence of Blastocystis subtypes within the Black Sea of Turkey. Copyright © 2018 Elsevier B.V. All rights reserved.

[Investigation of Coxiella burnetii contamination in commercial milk and PCR method for the detection of C. burnetii in egg].

PubMed

Hirai, Akihiko; Kaneko, Seiji; Nakama, Akiko; Ishizaki, Naoto; Odagiri, Megumi; Kai, Akemi; Sadamasu, Kenji; Shinkai, Takayuki; Yano, Kazuyoshi; Morozumi, Satoshi

2005-06-01

A total of 244 milk samples collected from supermarkets in Tokyo were examined for contamination with Coxiella burnetii. C. burnetii DNA was detected in 131 (53.7%) of the samples by nested PCR. PCR-positive samples were injected into immunosuppressed A/J strain mice. Of the 22 PCR-positive milk samples tested, none resulted in isolation of C. burnetii from the mice. Heat-treatment was sufficient to inactivate C. burnetii in commercial milk. In addition, a PCR detection method for C. burnetii in chicken egg was developed. Egg yolk was added to an equal volume of 1 mol/L of NaCl phosphate buffer and homogenized for removal of protein and lipid. After centrifugal separation, the supernatant was removed, and template DNA in the precipitate was extracted using SDS, proteinase K and NaI. Using such prepared samples, 3.2 x 10(1) C. burnetii particles in 1 g of egg yolk could be detected by nested PCR. All of 200 chicken egg samples collected from supermarkets in Tokyo were negative for C. burnetii by the nested PCR method.

Detection and quantification of Vibrio parahaemolyticus in shellfish from Italian production areas.

PubMed

Suffredini, Elisabetta; Mioni, Renzo; Mazzette, Rina; Bordin, Paola; Serratore, Patrizia; Fois, Federica; Piano, Annamaria; Cozzi, Loredana; Croci, Luciana

2014-08-01

Vibrio parahaemolyticus is a marine microorganism, recognized as an important cause of foodborne illness particularly in Asia, South America and United States. Outbreaks are rarely reported in Europe, but they can occur unexpectedly in relation, among other reasons, to the spread of highly virulent strains. It is known that the risk is proportional to exposure levels to pathogenic V. parahaemolyticus (i.e. carrying the tdh and/or the trh genes) but currently there is a lack of occurrence data for pathogenic V. parahaemolyticus in shellfish production areas of the Member States. In this study a total of 147 samples of bivalve molluscs, from harvesting areas of two Italian regions (Sardinia and Veneto) were analyzed for Escherichia coli and salmonella, according to Reg 2073/2005, and for detection and enumeration of total and toxigenic V. parahaemolyticus strains using a new DNA colony hybridization method. Environmental parameters (water temperature and salinity) were also recorded. Results of E. coli were consistently in agreement with the legislation limits for the harvesting class of origin and Salmonella was detected only in one sample. The average contamination levels for total V. parahaemolyticus were 84 and 73 CFU/g respectively for Sardinia and Veneto, with the highest value reaching 8.7 × 10(3)CFU/g. Nineteen samples (12.9%) resulted positive for the presence of potentially pathogenic V. parahaemolyticus strains, with levels ranging between 10 and 120 CFU/g and most of the positive samples (n=17) showing values equal or below 20 CFU/g. A significant correlation (r=0.41) was found between water temperature and V. parahaemolyticus levels, as well as with isolation frequency. The data provided in this study on contamination levels of total and potentially pathogenic V. parahaemolyticus, seasonal distribution and correlation with water temperature, will help in defining appropriate monitoring programs and post-harvest policies for this hazard, improving the management of the harvesting areas and the safety of bivalve molluscs. Copyright © 2014 Elsevier B.V. All rights reserved.

Microbiological quality assessment of milk at different stages of the dairy value chain in a developing country setting.

PubMed

Islam, Mohammad Aminul; Roy, Subarna; Nabi, Ashikun; Solaiman, Sultana; Rahman, Mahdia; Huq, Mohsina; Siddiquee, Nurul Amin; Ahmed, Niyaz

2018-08-02

The main objective of the study was to assess the microbiological quality of milk at different stages of the dairy value chain from farm to the factory in Bangladesh. A total of 438 raw milk samples (387 from primary producers, 32 from collectors, 15 from chilling plants, 4 from local restaurants) and 95 commercially processed milk samples were collected from northern part of Bangladesh. Almost 72% (n = 280) of samples at producer level and 100% from both collectors (n = 32) and chilling plants (n = 15) were contaminated with coliforms while 57% (n = 220) of samples from producers, 91% (n = 29) of samples from collectors and 100% (n = 15) from chilling plants were contaminated with fecal coliforms. Around 31% (n = 119) of samples from producers were positive for E. coli whereas >60% (n = 20) and 100% (n = 15) samples from collectors and chilling plants, respectively were positive for E. coli. One quarter of samples from collectors were positive for B. cereus and coagulase positive staphylococci and 33% (n = 5) of samples from chilling plants were positive for both of these microorganisms. In case of commercially processed milk, 77% (n = 46) and 37% (n = 22) of pasteurized milk samples had a high aerobic plate count (APC) (10 4  CFU/ml) and coliform count (>10 CFU/ml), respectively. None of the samples was positive for Shigella spp., Salmonella spp., and Campylobacter spp. Among 158 E. coli positive raw milk samples, 9% (n = 14) contained pathogenic E. coli, and enteroaggregative E. coli (EAEC) and Shiga-toxin producing E. coli (STEC) were found to be the predominant pathotypes. Of the 23 pathogenic E. coli identified from 14 samples based on their gene contents, >95% (n = 22) were resistant to at least one antibiotic and 13% (n = 3) of isolates were resistant to ≥3 classes of antibiotics. Several factors including the time of milking, hygiene practices of the producers, cow breed and amount of milk produced by the cow were found to be significantly associated with high APC of milk samples. In conclusion, both raw and commercially pasteurized milk are highly contaminated with fecal organisms. For intervention, more emphasis should be given at producer's level as microorganisms introduced to milk at this stage get the longest time for survival and multiplication. Copyright © 2018 Elsevier B.V. All rights reserved.

Brain structure mediates the association between height and cognitive ability.

PubMed

Vuoksimaa, Eero; Panizzon, Matthew S; Franz, Carol E; Fennema-Notestine, Christine; Hagler, Donald J; Lyons, Michael J; Dale, Anders M; Kremen, William S

2018-05-11

Height and general cognitive ability are positively associated, but the underlying mechanisms of this relationship are not well understood. Both height and general cognitive ability are positively associated with brain size. Still, the neural substrate of the height-cognitive ability association is unclear. We used a sample of 515 middle-aged male twins with structural magnetic resonance imaging data to investigate whether the association between height and cognitive ability is mediated by cortical size. In addition to cortical volume, we used genetically, ontogenetically and phylogenetically distinct cortical metrics of total cortical surface area and mean cortical thickness. Height was positively associated with general cognitive ability and total cortical volume and cortical surface area, but not with mean cortical thickness. Mediation models indicated that the well-replicated height-general cognitive ability association is accounted for by individual differences in total cortical volume and cortical surface area (highly heritable metrics related to global brain size), and that the genetic association between cortical surface area and general cognitive ability underlies the phenotypic height-general cognitive ability relationship.

Loop Mediated Isothermal Amplification (LAMP) for Embryo Sex Determination in Pregnant Women at Eight Weeks of Pregnancy.

PubMed

Almasi, Mohammad Amin; Almasi, Galavizh

2017-01-01

In human, SRY (sex-determining region of the Y chromosome) is the major gene for the testis-determining factor which is found in normal XY males and in the rare XX males, and it is absent in normal XX females and many XY females. There are several methods which can indicate a male genotype by the presence of the amplified product of SRY gene. The aim of this study was to identify the SRY gene for embryo sex determination in human during pregnancy using loop mediated isothermal amplification (LAMP) method. A total of 15 blood samples from pregnant women at eight weeks of pregnancy were collected, and Plasma DNA was extracted. LAMP assay was performed using DNA obtained for detection of SRY gene. Furthermore, colorimetric LAMP assay for rapid and easy detection of SRY gene was developed. LAMP results revealed that the positive reaction was highly specific only with samples containing XY chromosomes, while no amplification was found in samples containing XX chromosomes. A total of 15 blood samples from pregnant women were seven male embryos (46.6%) and eight female embryos (53.4%). All used visual components in the colorimetric assay could successfully make a clear distinction between positive and negative ones. The LAMP assay developed in this study is a valuable tool capable of monitoring the purity and detection of SRY gene for sex determination.

Evidence of the presence of the Zika virus in Mexico since early 2015.

PubMed

Díaz-Quiñonez, José Alberto; López-Martínez, Irma; Torres-Longoria, Belem; Vázquez-Pichardo, Mauricio; Cruz-Ramírez, Edith; Ramírez-González, José Ernesto; Ruiz-Matus, Cuitláhuac; Kuri-Morales, Pablo

2016-12-01

To assess the possible circulation of Zika virus (ZIKV) prior to the first documented case in Mexico, we reanalyzed the stored samples from the states of Veracruz and Yucatán, which were originally collected to test for dengue (DENV) and chikungunya (CHIKV) but were negative for these viruses despite the symptomatology. The samples were originally collected between the 30 and 46 epidemiological weeks (EW) when the ZIKV was not yet declared as a Public Health Emergency of International Concern (PHEIC). From the total 4016 negative samples, a total of one hundred samples, 50 from Veracruz (CHIK - DENV - ) and 50 from Yucatán (4 CHIK - DENV - and 46 CHIK - or DENV - ), were tested for Zika virus by using RT-PCR. Results showed that in Veracruz and Yucatán, 20 % (10/50) and 70 % (35/50) were, respectively, ZIKV positive, indicating unequivocally the presence of ZIKV at least since July 2015. We also tested non-confirmed suspect measles cases from early 2015 for ZIKV by RT-PCR. Remarkably in 11 Mexican states, 86 % (18/21) were positive with the earlier symptoms onset as early as May 2015. Finally, RT-PCR analyses on RNA extracted from Aedes aegypti mosquitoes captured from January to March 2015 showed the presence of ZIKV, strongly suggesting that the vector was already carrying the virus at the start of 2015.

The presence-absence coliform test for monitoring drinking water quality.

PubMed Central

Rice, E W; Geldreich, E E; Read, E J

1989-01-01

The concern for improved monitoring of the sanitary quality of drinking water has prompted interest in alternative methods for the detection of total coliform bacteria. A simplified qualitative presence-absence test has been proposed as an alternate procedure for detecting coliform bacteria in potable water. In this paper data from four comparative studies were analyzed to compare the recovery of total coliform bacteria from drinking water using the presence-absence test, the multiple fermentation tube procedure, and the membrane filter technique. The four studies were of water samples taken from four different geographic areas of the United States: Hawaii, New England (Vermont and New Hampshire), Oregon, and Pennsylvania. Analysis of the results of these studies were compared, based upon the number of positive samples detected by each method. Combined recoveries showed the presence-absence test detected significantly higher numbers of samples with coliforms than either the fermentation tube or membrane filter methods, P less than 0.01. The fermentation tube procedure detected significantly more positive samples than the membrane filter technique, P less than 0.01. Based upon the analysis of the combined data base, it is clear that the presence-absence test is as sensitive as the current coliform methods for the examination of potable water. The presence-absence test offers a viable alternative to water utility companies that elect to use the frequency-of-occurrence approach for compliance monitoring. PMID:2493663

Seroprevalence of hepatitis A virus antibody in a population aged 0-30 years in Shanghai, China: implications for hepatitis A immunization.

PubMed

Zhu, Y; Yuan, Z; Zhao, Q; Chen, G; Xu, B

2013-03-01

This study aimed to determine current seroprevalence of hepatitis A virus (HAV) antibody in subjects aged 0-30 years in Shanghai. A total of 5515 subjects were recruited through random clustering sampling in 2009. Basic clinical and demographic information and records of HAV vaccination were acquired by questionnaire interviews and review of immunization records. Serum samples were collected and tested for anti-HAV IgM and total anti-HAV. The overall seroprevalence of total anti-HAV was 52·9%. Subjects aged 20-24 years had the lowest (34·4%) anti-HAV seropositivity and subjects aged 5-9 years had the highest (74·6%) positivity. Seroprevalence was significantly greater in subjects from suburban areas than in subjects from urban areas. Subjects who had HAV immunizations had significantly greater seropositivity for total anti-HAV. Previous hepatitis A immunization policies had a significant impact on the presence of anti-HAV seroprevalence in a population aged 0-30 years from Shanghai.

The detection and recovery of Erysipelothrix spp. in meat and abattoir samples in Western Australia.

PubMed

Wang, Q; Fidalgo, S; Chang, B J; Mee, B J; Riley, T V

2002-01-01

To investigate the occurrence of Erysipelothrix rhusiopathiae and other Erysipelothrix spp. in abattoir and meat samples in Western Australia. Samples were collected from various parts of pig and sheep carcasses, as well as different sections of slaughtering line, pen soil and effluent. Previously evaluated culture methods were applied for the isolation of Erysipelothrix spp., in conjunction with phenotypic and genotypic detection and identification procedures. Of 109 samples from the two abattoirs, 35 (32.1%) were Erysipelothrix genus-specific PCR-positive. These came from swabs of animal exterior surfaces and joints, slaughtering areas, pig pen soil and abattoir effluent. Four samples (3.7%) from sheep arthritic joints and pig abattoir effluent were also E. rhusiopathiae species-specific PCR-positive. Of 123 carcass washing samples, 12 (9.8%) were genus-specific PCR-positive, and these came from all five kinds of meat samples tested, including beef, lamb, mutton, pork and chicken. Four of them (3.3%) were also species-specific PCR-positive. A total of 25 isolates was recovered from the samples, of which seven were identified as E. rhusiopathiae, seven were consistent with E. tonsillarum, and the remaining 11 were other species of Erysipelothrix. Erysipelothrix spp. can still be isolated and identified from specimens of animal origin with relative ease, provided that appropriate cultural and molecular procedures are used. Clinical microbiology laboratories may need to improve their diagnostic protocols. This study confirms that E. rhusiopathiae and other species of Erysipelothrix continue to colonize and contaminate farmed animals and animal products. Erysipelothrix infection still poses a potential threat to the economy of the farmed animal industry, as well as being a potential human public health hazard.

Two Years After the Schmallenberg Virus Epidemic in the Netherlands: Does the Virus still Circulate?

PubMed

Veldhuis, A M B; Mars, M H; Roos, C A J; van Wuyckhuise, L; van Schaik, G

2017-02-01

Two years after the introduction of the Schmallenberg virus in north-western Europe, it is unknown whether the virus is still circulating in countries that were the first to be confronted with it. When the population-level immunity declines in Europe, reintroduction or the re-emergence of SBV in Europe might eventually result in an outbreak of similar magnitude of that seen in 2011-2012. The Netherlands was part of the primary outbreak region of SBV in 2011. The aim of this study was to determine whether SBV circulated amongst dairy herds in the Netherlands in 2013, and if so, to which extent. For this purpose, the presence of SBV-specific antibodies in naive cattle was investigated. A total of 394 dairy farms were sampled between October and December 2013 by collecting five serum samples per herd. Antibodies were detected in 1.1% [95% confidence interval (CI): 0.7-1.7)] of the animals. All seropositive animals were single reactors per herd and were at least 8 months old at sampling. As these results were inconclusive in demonstrating freedom of SBV circulation, a more in-depth investigation was initiated to provide more insight: an additional sample of 20 youngstock within the same age category (including the five initially sampled animals) was collected from 17 of the 21 positive herds and tested for SBV-specific antibodies. This resulted in 9 antibody-positive test results of 316 samples. Again, the positive samples were single reactors within the sample obtained from each farm, which is unlikely given the characteristics of SBV. Therefore, assuming the single reactors as false-positive, this survey showed with 95% confidence that the maximum possible prevalence of herds with SBV circulation in the Netherlands was <1% in 2013. © 2015 Blackwell Verlag GmbH.

Genotoxicity and physicochemical characteristics of traffic-related ambient particulate matter.

PubMed

de Kok, Theo M; Hogervorst, Janneke G; Briedé, Jacco J; van Herwijnen, Marcel H; Maas, Lou M; Moonen, Edwin J; Driece, Hermen A; Kleinjans, Jos C

2005-08-01

Exposure to ambient particulate matter (PM) has been linked to several adverse health effects. Since vehicular traffic is a PM source of growing importance, we sampled total suspended particulate (TSP), PM(10), and PM(2.5) at six urban locations with pronounced differences in traffic intensity. The mutagenicity, DNA-adduct formation, and induction of oxidative DNA damage by the samples were studied as genotoxicological parameters, in relation to polycyclic aromatic hydrocarbon (PAH) levels, elemental composition, and radical-generating capacity (RGC) as chemical characteristics. We found pronounced differences in the genotoxicity and chemical characteristics of PM from the various locations, although we could not establish a correlation between traffic intensity and any of these characteristics for any of the PM size fractions. Therefore, the differences between locations may be due to local sources of PM, other than traffic. The concentration of total (carcinogenic) PAHs correlated positively with RGC, direct and S9-mediated mutagenicity, as well as the induction of DNA adducts and oxidative DNA damage. The interaction between total PAHs and transition metals correlated positively with DNA-adduct formation, particularly from the PM(2.5) fraction. RGC was not associated with one specific PM size fraction, but mutagenicity and DNA reactivity after metabolic activation were relatively high in PM(10) and PM(2.5), when compared with TSP. We conclude that the toxicological characteristics of urban PM samples show pronounced differences, even when PM concentrations at the sample sites are comparable. This implies that emission reduction strategies that take chemical and toxicological characteristics of PM into account may be useful for reducing the health risks associated with PM exposure. Copyright 2005 Wiley-Liss, Inc.

Molecular detection of Theileria spp. and Babesia spp. in sheep and ixodid ticks from the northeast of Iran.

PubMed

Razmi, Gholamreza; Pourhosseini, Moslem; Yaghfouri, Saeed; Rashidi, Ahmad; Seidabadi, Mohsen

2013-02-01

Theilerioses and babesioses are important diseases in Iranian sheep. The present study was undertaken to identify and classify/specify Theileria spp. and Babesia spp. in sheep and vector ticks. Investigation was carried out from 2009 to 2011 in the Khorasan Razavi Province, Iran. In total, 302 sheep originating from 60 different flocks were clinically examined and their blood collected. In addition, from the same flocks, ixodid ticks were sampled. Stained blood smears were microscopically examined for the presence of Theileria and Babesia organisms, and a semi-nested PCR was used for subsequent molecular specification. From the ticks, salivary glands and uterus were isolated and subsequently analyzed by semi-nested PCR. Piroplasm organisms were observed in 29% of the blood smears with low parasitemia, whereas 65% of the blood samples yielded positive PCR findings. The presence of Theileria ovis (55.6%), Theileria lestoquardi, and mixed infection with Theileria spp. and Babesia ovis were detected by semi-nested PCR in 0.3%, 5.6%, and 0.99%, respectively. In total, 429 ixodid ticks were collected from different areas of the province. The most prevalent ticks were Rhipicephalus turanicus (n = 376; 87.6% of the total), followed by Hyalomma marginatum turanicum (n = 30; 7.0%), Dermacentor raskemensis (n = 12; 2.8%), Hyalomma anatolicum anatolicum (n = 7; 1.6%), Dermacentor marginatus (n = 2; 0.5%), Rhipicephalus bursa (n = 1; 0.2%), and Haemaphysalis sp. (n = 1; 0.2%). Of the positive R. turanicus samples, 5 (5.7%) were infected with T. ovis and 2 (2.9%) with T. lestoquardi. Neither Babesia ovis nor Babesia motasi infection was detected in salivary glands or uterine samples of the ticks. The results also suggest that R. turanicus could be the vector responsible for transmission of the 2 Theileria species.

Can direct gas-liquid chromatography of clinical samples detect specific organisms?

PubMed Central

Watt, B; Geddes, P A; Greenan, O A; Napier, S K; Mitchell, A

1982-01-01

A total of 1929 samples was analyzed by direct gas-liquid chromatography and the volatile fatty acid (VFA) patterns of the positive samples were compared with the results of culture. There was no correlation between any bacterial genus or species and the detailed VFA patterns although the presence of butyric or valeric acids, or both, was generally associated with the presence of anaerobes and that of acetic acid was generally associated with aerobic bacteria; however, the technique could not predict the nature of the subsequent bacterial isolate. There was also poor correlation between the VFA pattern in a given sample and the VFA pattern(s) of anaerobic bacteria subsequently isolated from that sample. PMID:7096589

Passive surveillance of Leptospira infection in swine in Germany.

PubMed

Strutzberg-Minder, Katrin; Tschentscher, Astrid; Beyerbach, Martin; Homuth, Matthias; Kreienbrock, Lothar

2018-01-01

As no current data are available on the prevalence of leptospiral infection in swine in Germany, we analysed laboratory data from diagnostic examinations carried out on samples from swine all over Germany from January 2011 to September 2016. A total of 29,829 swine sera were tested by microscopic agglutination test (MAT) for antibodies against strains of eleven Leptospira serovars. Overall, 20.2% (6025) of the total sample collection tested positive for leptospiral infection. Seropositivity ranged between 16.3% (964) in 2011 and 30.9% (941) in 2016 (January to September only). Of all samples, 11.6% (57.3% of the positives) reacted with only one Leptospira serovar, and only 8.6% (42.7% of the positives) reacted simultaneously with two or more serovars. The most frequently detected serovar was Bratislava, which was found in 11.6% (3448) of all samples, followed by the serovars Australis in 7.3% (2185), Icterohaemorrhagiae in 4.0% (1191), Copenhageni in 4.0% (1182), Autumnalis in 3.7% (1054), Canicola in 2.0% (585), and Pomona in 1.2% (368). Modelling shows that both the year and the reason for testing at the laboratory had statistically strong effects on the test results; however, no interactions were determined between those factors. The results support the suggestion that the seropositivities found may be considered to indicate the state of leptospiral infections in the German swine population. Although data from passive surveillance are prone to selection bias, stratified analysis by initial reason for examination and analyses by model approaches may correct for biases. A prevalence of about 20% for a leptospiral infection is most probable for sows with reproductive problems in Germany, with an increasing trend. Swine in Germany are probably a reservoir host for serovar Bratislava, but in contrast to other studies not for Pomona and Tarassovi.

Frequency of tuberculous and non-tuberculous mycobacteria in HIV infected patients from Bogota, Colombia

PubMed Central

Murcia-Aranguren, Martha I; Gómez-Marin, Jorge E; Alvarado, Fernando S; Bustillo, José G; de Mendivelson, Ellen; Gómez, Bertha; León, Clara I; Triana, William A; Vargas, Erwing A; Rodríguez, Edgar

2001-01-01

Background The prevalence of infections by Mycobacterium tuberculosis and non-tuberculous Mycobacterium species in the HIV-infected patient population in Colombia was uncertain despite some pilot studies. We determined the frequency of isolation of Mycobacterium tuberculosis and of non-tuberculous Mycobacterium species in diverse body fluids of HIV-infected patients in Bogota, Colombia. Methods Patients who attended the three major HIV/AIDS healthcare centres in Bogota were prospectively studied over a six month period. A total of 286 patients were enrolled, 20% of them were hospitalized at some point during the study. Sixty four percent (64%) were classified as stage C, 25% as stage B, and 11% as stage A (CDC staging system, 1993). A total of 1,622 clinical samples (mostly paired samples of blood, sputum, stool, and urine) were processed for acid-fast bacilli (AFB) stain and culture. Results Overall 43 of 1,622 cultures (2.6%) were positive for mycobacteria. Twenty-two sputum samples were positive. Four patients were diagnosed with M. tuberculosis (1.4%). All isolates of M. tuberculosis were sensitive to common anti-tuberculous drugs. M. avium was isolated in thirteen patients (4.5%), but only in three of them the cultures originated from blood. The other isolates were obtained from stool, urine or sputum samples. In three cases, direct AFB smears of blood were positive. Two patients presented simultaneously with M. tuberculosis and M. avium. Conclusions Non-tuberculous Mycobacterium infections are frequent in HIV infected patients in Bogota. The diagnostic sensitivity for infection with tuberculous and non-tuberculous mycobacteria can be increased when diverse body fluids are processed from each patient. PMID:11722797

Quantification of γ-Aminobutyric Acid in Cerebrospinal Fluid Using Liquid Chromatography-Electrospray Tandem Mass Spectrometry.

PubMed

Arning, Erland; Bottiglieri, Teodoro

2016-01-01

We describe a simple stable isotope dilution method for accurate and precise measurement of γ-aminobutyric acid (GABA), a major inhibitory neurotransmitter in human cerebrospinal fluid (CSF) as a clinical diagnostic test. Determination of GABA in CSF (50 μL) was performed utilizing high performance liquid chromatography coupled with electrospray positive ionization tandem mass spectrometry (HPLC-ESI-MS/MS). Analysis of free and total GABA requires two individual sample preparations and mass spectrometry analyses. Free GABA in CSF is determined by a 1:2 dilution with internal standard (GABA-D2) and injected directly onto the HPLC-ESI-MS/MS system. Determination of total GABA in CSF requires additional sample preparation in order to hydrolyze all the bound GABA in the sample to the free form. This requires hydrolyzing the sample by boiling in acidic conditions (hydrochloric acid) for 4 h. The sample is then further diluted 1:10 with a 90 % acetonitrile/0.1 % formic acid solution and injected into the HPLC-ESI-MS/MS system. Each assay is quantified using a five-point standard curve and is linear from 6 nM to 1000 nM and 0.63 μM to 80 μM for free and total GABA, respectively.

A consensus prognostic gene expression classifier for ER positive breast cancer

PubMed Central

Teschendorff, Andrew E; Naderi, Ali; Barbosa-Morais, Nuno L; Pinder, Sarah E; Ellis, Ian O; Aparicio, Sam; Brenton, James D; Caldas, Carlos

2006-01-01

Background A consensus prognostic gene expression classifier is still elusive in heterogeneous diseases such as breast cancer. Results Here we perform a combined analysis of three major breast cancer microarray data sets to hone in on a universally valid prognostic molecular classifier in estrogen receptor (ER) positive tumors. Using a recently developed robust measure of prognostic separation, we further validate the prognostic classifier in three external independent cohorts, confirming the validity of our molecular classifier in a total of 877 ER positive samples. Furthermore, we find that molecular classifiers may not outperform classical prognostic indices but that they can be used in hybrid molecular-pathological classification schemes to improve prognostic separation. Conclusion The prognostic molecular classifier presented here is the first to be valid in over 877 ER positive breast cancer samples and across three different microarray platforms. Larger multi-institutional studies will be needed to fully determine the added prognostic value of molecular classifiers when combined with standard prognostic factors. PMID:17076897

Effect of illite particle shape on cesium sorption

USGS Publications Warehouse

Rajec, Pavol; Šucha, Vladimír; Eberl, Dennis D.; Środoń, Jan; Elsass, Françoise E.

1999-01-01

Samples containing illite and illite-smectite, having different crystal shapes (plates, “barrels”, and filaments), were selected for sorption experiments with cesium. There is a positive correlation between total surface area and Cs-sorption capacity, but no correlation between total surface area and the distribution coefficient, Kd. Generally Kd increases with the edge surface area, although “hairy” (filamentous) illite does not fit this pattern, possibly because elongation of crystals along one axis reduces the number of specific sorption sites.

Fears of happiness and compassion in relationship with depression, alexithymia, and attachment security in a depressed sample.

PubMed

Gilbert, Paul; McEwan, Kirsten; Catarino, Francisca; Baião, Rita; Palmeira, Lara

2014-06-01

In a non-clinical population, fears of compassion and fear of happiness have both been found to be highly correlated with alexithymia and depression. This study sought to explore these processes and their links with adult attachment and social safeness and pleasure in a depressed group. A total of 52 participants suffering from moderate to severe depression completed measures of fears of happiness, compassion from others and for self, in addition to measures of alexithymia, attachment, social safeness, and depression, anxiety, and stress. Fears of compassion and happiness were highly correlated with alexithymia, adult attachment, and depression, anxiety, and stress. Fear of happiness was found to be the best predictor of depression, anxiety, and stress, whereas fear of compassion from others was the best predictor of adult attachment. A path analysis showed that fears of positive emotion fully mediate the link between alexithymia and depression. This clinical sample had higher mean scores in fears of positive emotions, alexithymia, and depression, anxiety, and stress than a previously studied student sample. This study adds to the evidence that fears of positive emotions are important features of mental health difficulties. Unaddressed, these fears can block positive emotions and may lead to emotional avoidance of positive affect thus contributing as blocks to successful therapy. Therapies for depression may therefore profitably assess and desensitize the fear of positive emotions. © 2013 The British Psychological Society.

Molecular Detection and Genotyping of Chlamydia psittaci in Captive Psittacines from Costa Rica.

PubMed

Sheleby-Elías, Jessica; Solórzano-Morales, Antony; Romero-Zuñiga, Juan José; Dolz, Gaby

2013-01-01

Oropharyngeal and cloacal swabs from 117 captive psittacine birds presented at veterinary clinics (88) and from shelters/rescue centers of wildlife (29) were collected to determine the prevalence of C. psittaci in captive birds in Costa Rica. Samples were collected during 2009 from a total of 19 different species of parrots, with Ara macao (33), Amazona autumnalis (24), Amazona ochrocephala (21), and Ara ararauna (8) being the most representative species sampled. C. psittaci was detected in four (3.4%) birds using molecular detection (PCR). The positive samples belonged to birds presented at veterinary clinics; three of them were Ara macao and one Amazona ochrocephala. Three birds were adults; all positive birds showed no symptoms of illness and lived in homes with other birds, two in San José and two in Heredia. Sequencing was used to confirm the PCR positive results, showing that two samples of C. psittaci belonged to genotype A, representing the first report of the presence of this genotype in Costa Rica. The detection of this bacterium in captive psittacine birds shows that there is a potential risk for people living or having contact with them and that there is a possibility of infecting other birds.

Molecular Detection and Genotyping of Chlamydia psittaci in Captive Psittacines from Costa Rica

PubMed Central

Sheleby-Elías, Jessica; Solórzano-Morales, Ántony; Romero-Zuñiga, Juan José

2013-01-01

Oropharyngeal and cloacal swabs from 117 captive psittacine birds presented at veterinary clinics (88) and from shelters/rescue centers of wildlife (29) were collected to determine the prevalence of C. psittaci in captive birds in Costa Rica. Samples were collected during 2009 from a total of 19 different species of parrots, with Ara macao (33), Amazona autumnalis (24), Amazona ochrocephala (21), and Ara ararauna (8) being the most representative species sampled. C. psittaci was detected in four (3.4%) birds using molecular detection (PCR). The positive samples belonged to birds presented at veterinary clinics; three of them were Ara macao and one Amazona ochrocephala. Three birds were adults; all positive birds showed no symptoms of illness and lived in homes with other birds, two in San José and two in Heredia. Sequencing was used to confirm the PCR positive results, showing that two samples of C. psittaci belonged to genotype A, representing the first report of the presence of this genotype in Costa Rica. The detection of this bacterium in captive psittacine birds shows that there is a potential risk for people living or having contact with them and that there is a possibility of infecting other birds. PMID:24163776

Rapid Detection of Staphylococcus aureus and Methicillin-Resistant S. aureus in Atopic Dermatitis by Using the BD Max StaphSR Assay.

PubMed

Lee, Mi Kyung; Park, Kui Young; Jin, Taewon; Kim, Ju Hee; Seo, Seong Jun

2017-07-01

Eczematous lesions of atopic dermatitis (AD) patients are known to be a source of Staphylococcus aureus (SA) transmission and might be a reservoir for community-associated methicillin-resistant SA (MRSA). The BD Max StaphSR (BD-SR) is a fully automated, multiplex real-time PCR assay for the direct detection and differentiation of SA and MRSA from nasal swab samples. We evaluated the detection rates of SA and MRSA from skin lesions of outpatients with AD using the BD-SR assay, and determined the usefulness of the BD-SR assay. A total of 244 skin swab samples (skin lesions of 213 outpatients with AD and normal skin of 31 healthy controls) were tested directly by using the BD-SR assay. Of the 213 samples from patients with AD, 69 (32.4%) were positive for SA, 6 (8.7%) of which were positive for MRSA. Only 1 (3.2%) of 31 samples from healthy controls was positive for SA. The BD-SR assay is effective for the rapid detection of SA and MRSA from skin swab samples, which can provide important information for managing patients with AD and preventing the spread of MRSA. © The Korean Society for Laboratory Medicine.

Effects on goat milk quality of the presence of Mycoplasma spp. in herds without symptoms of contagious agalactia.

PubMed

de la Fe, Christian; Sánchez, Antonio; Gutierrez, Aldo; Contreras, Antonio; Carlos Corrales, Juan; Assunçao, Patricia; Poveda, Carlos; Poveda, José B

2009-02-01

This study was designed to assess the possible effects of mycoplasmas on the quality of milk produced by goat herds in a contagious agalactia (CA) endemic area with absence of classical symptoms. Several factors related to milk quality (percentages of fat, total protein, lactose and total solids, standard plate counts (SPC) and presence of Staphylococcus aureus) were compared in mycoplasma-infected and non-infected herds. To define the CA status of 26 herds on the island of Lanzarote (Spain), where CA is endemic, 570 individual milk samples and 266 bulk tank milk (BTM) samples were microbiologically analysed for the presence of Mycoplasma spp. A herd was considered infected by mycoplasmas when at least a sample (individual or BTM) was positive. BTM samples were also used to determine milk quality parameters. Mycoplasma infection was confirmed in 13 herds. A total of 31, 10 and 11 strains of Mycoplasma mycoides subsp. mycoides LC (MmmLC), Mp. agalactiae and Mp. capricolum subsp. capricolum were isolated. No significant differences were observed between the least square means of the variables fat, total protein, lactose and total solids or SPC recorded for the infected v. non-infected herds. The Staph. aureus status of a herd was also found to be independent of the presence of Mycoplasma spp. Our findings indicate that neither the presence of mycoplasmas in a goat herd with absence of classical symptoms seem to compromise the quality of the BTM.

[Levels of Ochratoxin A and total Aflatoxins in Panamanian exportation coffee by an ELISA Method].

PubMed

Franco, Heriberto; Vega, Aracelly; Reyes, Stephany; De Léon, Javier; Bonilla, Alexis

2014-03-01

A study about processing conditions of exportation coffee in 15 benefits located in Chiriqui, western region of Panama, was conducted. In addition, 21 samples of processed coffee (green beans), from the benefits, were analyzed. The samples were microbiologically tested in order to quantify total aflatoxins (B1, B2, G1 and G2) and Ochratoxin A (OTA), using the immunoaffinity ELISA method. A detection limit of 0.017 ng/mL, was determined for Ochratoxin A, which is equivalent to a concentration of 0.829 µg/kg, and a detection limit of 0.027 ng/mL, for total aflatoxins, which is equivalent to a concentration of 1.350 µg/kg. It was found that four (19%) out of the 21 samples were positive to the presence of Ochratoxin A and three (14%) to the presence of total aflatoxins. Samples showed levels of Ochratoxin A in the range 4.90 - 37.73 µg/kg; only three of them exceeded the maximum limit allowed by the European Union, for the concentration of Ochratoxin, which is of 5.0 µg/kg. Total aflatoxins were found in the range 1.51 - 1.93 µg/kg, below 10 µg/kg which is the maximum limit allowed for coffee by the European Union. The results indicate that the processing of coffee produced in Panama successfully meets international standards for postharvest handling, which leads to a low incidence ofmycotoxins and very low levels ofmycotoxin-producing fungi.

[Correlation analysis of nutrients and microorganisms in soils with polyphenols and total flavonoids of Houttuynia cordata].

PubMed

Wu, Dan; Luo, Shi-qiong; Yang, Zhan-nan; Ma, Jing; Hong, Liang

2015-04-01

The relationship of nutrients and microorganisms in soils with polyphenols and total flavonoids of Houttuynia cordata were investigated by measuring nutrients, enzyme activity, pH, concentrations of microbe phospholipid fatty acids (PLFAs) in soils, and determining concentrations of polyphenols and total flavonoids of H. cordata. The research is aimed to understand characteristics of the planting soils and improve the quality of cultivated H. cordata. The soils at different sample sites varied greatly in nutrients, enzyme activity, pH, microbic PLFAs and polyphenols and all flavonoids. The content of total PLFAs in sample sites was following: bacteria > fungi > actinomyces > nematode. The content of bacteria PLFAs was 37.5%-65.0% at different sample sites. Activities of polyphenol oxidease, concentrations of available P and content of PLFAs of bacteria, actinomyces and total microorganisms in soils were significantly and positively related to the concentrations of polyphenols and total flavonoids of H. cordata, respectively (P < 0.05) . The Content of fungi PLFAs in soils was significantly and negatively related to concentrations of polyphenols and total flavonoids of H. cordata, respectively (P < 0.05). This study provides evidence that effectiveness of the soil nutrient, which may be improved due to transformation of soil microorganisms and enzymes to N and P in the soils, was beneficial to adaptation of H. cordata adapted to different soil conditions, and significantly affects metabolic accumulation of polyphenols and flavonoids of H. cordata.

High hepatitis E virus antibody positive rates in dogs and humans exposed to dogs in the south-west of China.

PubMed

Zeng, M Y; Gao, H; Yan, X X; Qu, W J; Sun, Y K; Fu, G W; Yan, Y L

2017-12-01

Hepatitis E (HE) is a zoonotic viral disease caused by hepatitis E virus (HEV). The objective of this study was to investigate the prevalence of HEV infection among dogs and humans exposed to dogs in the south-west region of China. A total of 4,490 dog serum samples and 2,206 relative practitioner serum samples were collected from 18 pet hospitals and dog farms in Yunnan, Sichuan and Guizhou province, and the anti-HEV IgG antibodies were detected by ELISA. The results showed that the total positive rate of anti-HEV antibodies was 36.55% with the highest rate in city stray dogs, and the differences in distinct species and growth phases were significant. The positive rate of anti-HEV antibody in veterinarian and farm staff-related practitioners was significantly higher than the general population. The finding of the present survey suggested that high HEV seroprevalence in dogs and humans exposed to dogs in the south-west area of China poses a significant public health concern. It is urgent to improve integrated strategies to detect, prevent and control HEV infection in dogs and humans exposed to dogs in this area. © 2017 Blackwell Verlag GmbH.

Epidemiology of intramammary infections with Staphylococcus aureus and mastitis streptococci in a dairy cattle herd with a history of recurrent clinical mastitis.

PubMed

Vlkova, H; Babak, V; Vrtkova, I; Cervinkova, D; Marosevic, D; Moravkova, M; Jaglic, Z

2017-03-28

The aim of the present work was to examine a dairy herd with an anamnesis of recurrent clinical mastitis and decreased milk production. A total of 239 individual cow milk samples originating from asymptomatic cows were collected at four-month intervals and examined mainly for the presence of Staphylococcus aureus and mastitis streptococci using standard cultivation methods. In total, 29.7% and 9.2% samples were positive for S. aureus and mastitis streptococci, respectively. Unlike for mastitis streptococci, the prevalence of animals positive for S. aureus had an increasing trend (p<0.05; Chi-squared test for trend) with rising parity. Despite in vitro susceptibility of S. aureus to potentiated penicillins and cephalosporins, the persistence of S. aureus was observed in cows undergoing intramammary treatment with amoxicillin/clavulanic acid (a potentiated penicillin antibiotic). All isolates of S. aureus were biofilm-positive and had the same macrorestriction pattern. Furthermore, no dependence was observed between the occurrence of S. aureus in milk and previous cases of clinical mastitis, reproductive and periparturient disorders and administration of antibiotics. In contrast to S. aureus, the occurrence of mastitis streptococci in milk was linked with previous cases of clinical mastitis and intramammary administration of antibiotics.

The pharmacokinetics of abacavir 600 mg once daily in HIV-1-positive pregnant women.

PubMed

Schalkwijk, Stein; Colbers, Angela; Konopnicki, Deborah; Weizsäcker, Katharina; Moltó, José; Tenorio, Carmen Hidalgo; Hawkins, David; Taylor, Graham; Wood, Chris; van der Ende, Marchina; Burger, David

2016-05-15

To describe the pharmacokinetics of abacavir 600 mg once daily (q.d.) in HIV-1-positive women during pregnancy and postpartum. A nonrandomized, open-label, multicentre, phase-IV study. HIV-positive pregnant women receiving abacavir 600 mg q.d. as part of clinical care were included. Intensive 24-h pharmacokinetic sampling was performed during the third trimester and at least 2 weeks after delivery. Pharmacokinetic parameters were calculated by noncompartmental analysis. Paired cord blood and maternal blood samples were taken at delivery when feasible. A total of 14 women were included in the analysis. Geometric mean ratios (90% confidence intervals) of third trimester versus postpartum were 1.05 (0.92-1.19) for AUC0-24h and 1.00 (0.83-1.21) for Cmax. The median (range) ratio of abacavir cord plasma to maternal plasma was 1.0 (0.7-1.0, n = 3). Viral load at the third trimester visit was less than 50 copies/ml in 13 participants (93%; one unknown). In total, 13 (93%; one unknown) children were tested HIV-negative. The pharmacokinetics of abacavir 600 mg q.d. during pregnancy are equivalent to postpartum. No dose adjustments are required during pregnancy and similar antiviral activity is expected.

Applying Incremental Sampling Methodology to Soils Containing Heterogeneously Distributed Metallic Residues to Improve Risk Analysis.

PubMed

Clausen, J L; Georgian, T; Gardner, K H; Douglas, T A

2018-01-01

This study compares conventional grab sampling to incremental sampling methodology (ISM) to characterize metal contamination at a military small-arms-range. Grab sample results had large variances, positively skewed non-normal distributions, extreme outliers, and poor agreement between duplicate samples even when samples were co-located within tens of centimeters of each other. The extreme outliers strongly influenced the grab sample means for the primary contaminants lead (Pb) and antinomy (Sb). In contrast, median and mean metal concentrations were similar for the ISM samples. ISM significantly reduced measurement uncertainty of estimates of the mean, increasing data quality (e.g., for environmental risk assessments) with fewer samples (e.g., decreasing total project costs). Based on Monte Carlo resampling simulations, grab sampling resulted in highly variable means and upper confidence limits of the mean relative to ISM.

Evaluation of TECRA broth, Bolton broth, and direct plating for recovery of Campylobacter spp, from broiler carcass rinsates from commercial processing plants.

PubMed

Richardson, L J; Cox, N A; Bailey, J S; Berrang, M E; Cox, J M; Buhr, R J; Fedorka-Cray, P J; Harrison, M A

2009-05-01

The purpose of this study was to compare a conventional culture broth method (Bolton enrichment), a newly developed proprietary broth method (TECRA Campylobacter enrichment), and direct plating for recovery of Campylobacter spp. from chicken carcass rinsates. Whole carcass rinses were taken from 140 carcasses at rehang (immediately after defeathering but before evisceration) and from 140 carcasses at postchill from eight different processing plants in the United States. The rinsate samples were packed in ice and shipped overnight to the laboratory. Aliquots of the rinsate were transferred into Bolton and TECRA enrichment broths and were direct plated. Standard laboratory procedures with Campy-cefex plates were followed for recovery of Campylobacter spp. For rehang carcasses, 94% were positive for Campylobacter spp. with the TECRA enrichment broth and 74% were positive with the Bolton enrichment broth. For postchill carcasses, 74% were positive for Campylobacter spp. with the TECRA enrichment broth and 71% were positive with the Bolton enrichment broth. Compared with the Bolton enrichment broth, TECRA enrichment broth significantly suppressed non-Campylobacter microflora (P < 0.05). Overall, TECRA enrichment broth yielded an 11% higher total number of Campylobacter-positive samples compared with the Bolton enrichment broth. Campylobacter spp. detection in postchill samples was significantly greater (P < 0.05) by enrichment (84%) than by direct plating (19%). The high number of Campylobacter-positive samples obtained with all procedures indicated that 99% of the carcass rinsates obtained at rehang and 84% obtained at postchill contained Campylobacter spp.

Detection of antibiotics in chicken eggs obtained from supermarkets in Ho Chi Minh City, Vietnam.

PubMed

Yamaguchi, Takahiro; Okihashi, Masahiro; Harada, Kazuo; Konishi, Yoshimasa; Uchida, Kotaro; Hoang Ngoc Do, Mai; Thi Bui, Long; Nguyen, Thinh Duc; Phan, Ha Bich; Dang Thien Bui, Huong; Nguyen, Phuc Do; Kajimura, Keiji; Kumeda, Yuko; Van Dang, Chinh; Hirata, Kazumasa; Yamamoto, Yoshimasa

2017-06-03

The residual levels of antibiotics in Vietnamese eggs were monitored from 2014 to 2015. A total of 111 egg packages, distributed by 11 different companies, were collected from supermarkets in Ho Chi Minh City and the levels of 28 antibiotics were analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) screening method. Sixteen samples tested positive for antibiotics; a total of eight compounds (enrofloxacin, ciprofloxacin, norfloxacin, sulfadimethoxine, sulfamethazine, sulfamonomethoxine, tilmicosin and trimethoprim) were detected. Enrofloxacin was detected in eight samples, with two samples exhibiting concentrations exceeding 1,000 µg kg -1 . Tilmicosin was detected in three samples at a range of 49-568 µg kg -1 . We observed that two of the 11 companies frequently sold antibiotic-contaminated eggs (detection rates of 56 and 60%), suggesting that a number of companies do not regulate the use of antibiotics in egg-laying hens. Our findings indicate that livestock farmers require instruction regarding antibiotic use and that continual antibiotic monitoring is essential in Vietnam.

[Hygienic evaluation of the total mutagenic activity of snow samples from Magnitogorsk].

PubMed

Legostaeva, T B; Ingel', F I; Antipanova, N A; Iurchenko, V V; Iuretseva, N A; Kotliar, N N

2010-01-01

The paper gives the results of 4-year monitoring of the total mutagenic activity of snow samples from different Magnitogork areas in a test for induction of dominant lethal mutations (DLM) in the gametes of Drosophila melanogaster. An association was first found between the rate of DLM and the content of some chemical compounds in the ambient air and snow samples; moreover all the substances present in the samples, which had found genotoxic effects, showed a positive correlation with the rate of DLM. Furthermore, direct correlations were first established between the rate of DLM and the air pollution index and morbidity rates in 5-7-year-old children residing in the areas under study. The findings allow the test for induction of dominant lethal mutations (DLM) in the gametes of Drosophila melanogaster to be recommended due to its unique informative and prognostic value for monitoring ambient air pollution and for extensive use in the risk assessment system.

Environmental assessment of three egg production systems - Part III: Airborne bacteria concentrations and emissions.

PubMed

Zhao, Y; Zhao, D; Ma, H; Liu, K; Atilgan, A; Xin, H

2016-07-01

Airborne microorganism level is an important indoor air quality indicator, yet it has not been well documented for laying-hen houses in the United States. As a part of the Coalition for Sustainable Egg Supply (CSES) environmental monitoring project, this study comparatively monitored the concentrations and emissions of airborne total and Gram-negative (Gram(-)) bacteria in three types of commercial laying-hen houses, i.e., conventional cage (CC), aviary (AV), and enriched colony (EC) houses, over a period of eight months covering the mid and late stages of the flock cycle. It also delineated the relationship between airborne total bacteria and particulate matter smaller than 10 μm in aerodynamic diameter (PM10). The results showed airborne total bacteria concentrations (log CFU/m(3)) of 4.7 ± 0.3 in CC, 6.0 ± 0.8 in AV, and 4.8 ± 0.3 in EC, all being higher than the level recommended for human environment (3.0 log CFU/m(3)). The much higher concentrations in AV arose from the presence of floor litter and hen activities on it, as evidenced by the higher concentrations in the afternoon (with litter access) than in the morning (without litter access). The overall means and standard deviation of airborne total bacteria emission rates, in log CFU/[h-hen] (or log CFU/[h-AU], AU = animal unit or 500 kg live weight) were 4.8 ± 0.4 (or 7.3 ± 0.4) for CC, 6.1 ± 0.7 (or 8.6 ± 0.7) for AV, and 4.8 ± 0.5 (or 7.3 ± 0.5) for EC. Both concentration and emission rate of airborne total bacteria were positively related to PM10 Gram(-) bacteria were present at low concentrations in all houses; and only 2 samples (6%) in CC, 7 (22%) samples in AV, and 2 (6%) samples in EC out of 32 air samples collected in each house were found positive with Gram(-) bacteria. The concentration of airborne Gram(-) bacteria was estimated to be <2% of the total bacteria. Total bacteria counts in manure on belt (in all houses) and floor litter (only in AV) were similar; however, the manure had much more Gram(-) bacteria than the litter. The results point out the need to mitigate airborne total bacteria in laying-hen houses, especially in AV houses. © 2016 Poultry Science Association Inc.

Application of enzyme-linked immunosorbent assay for measurement of polychlorinated biphenyls from hydrophobic solutions: Extracts of fish and dialysates of semipermeable membrane devices: Chapter 26

USGS Publications Warehouse

Zajicek, James L.; Tillitt, Donald E.; Huckins, James N.; Petty, Jimmie D.; Potts, Michael E.; Nardone, David A.

1996-01-01

Determination of PCBs in biological tissue extracts by enzyme-linked immunosorbent assays (ELISAs) can be problematic, since the hydrophobic solvents used for their extraction and isolation from interfering biochemicals have limited compatibility with the polar solvents (e.g. methanol/water) and the immunochemical reagents used in ELISA. Our studies of these solvent effects indicate that significant errors can occur when microliter volumes of PCB containing extracts, in hydrophobic solvents, are diluted directly into methanol/water diluents. Errors include low recovery and excess variability among sub-samples taken from the same sample dilution. These errors are associated with inhomogeneity of the dilution, which is readily visualized by the use of a hydrophobic dye, Solvent Blue 35. Solvent Blue 35 is also used to visualize the evaporative removal of hydrophobic solvent and the dissolution of the resulting PCB/dye residue by pure methanol and 50% (v/v) methanol/water, typical ELISA diluents. Evaporative removal of isooctane by an ambient temperature nitrogen purge with subsequent dissolution in 100% methanol gives near quantitative recovery of model PCB congeners. We also compare concentrations of total PCBs from ELISA (ePCB) to their corresponding concentrations determined from capillary gas chromatography (GC) in selected fish sample extracts and dialysates of semipermeable membrane device (SPMD) passive samplers using an optimized solvent exchange procedure. Based on Aroclor 1254 calibrations, ePCBs (ng/mL) determined in fish extracts are positively correlated with total PCB concentrations (ng/mL) determined by GC: ePCB = 1.16 * total-cPCB - 5.92. Measured ePCBs (ng/3 SPMDs) were also positively correlated (r2 = 0.999) with PCB totals (ng/3 SPMDs) measured by GC for dialysates of SPMDs: ePCB = 1.52 * total PCB - 212. Therefore, this ELISA system for PCBs can be a rapid alternative to traditional GC analyses for determination of PCBs in extracts of biota or in SPMD dialysates.

Generation and distribution of PAHs in the process of medical waste incineration

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen, Ying, E-mail: echochen327@163.com; National Center of Solid Waste Management, Ministry of Environmental Protection, Beijing 100029; Zhao, Rongzhi

Highlights: ► PAHs generation and distribution features of medical waste incineration are studied. ► More PAHs were found in fly ash than that in bottom ash. ► The highest proportion of PAHs consisted of the seven most carcinogenic ones. ► Increase of free oxygen molecule and burning temperature promote PAHs degradation. ► There is a moderate positive correlation between total PCDD/Fs and total PAHs. - Abstract: After the deadly earthquake on May 12, 2008 in Wenchuan county of China, several different incineration approaches were used for medical waste disposal. This paper investigates the generation properties of polycyclic aromatic hydrocarbons (PAHs)more » during the incineration. Samples were collected from the bottom ash in an open burning slash site, surface soil at the open burning site, bottom ash from a simple incinerator, bottom ash generated from the municipal solid waste (MSW) incinerator used for medical waste disposal, and bottom ash and fly ash from an incinerator exclusively used for medical waste. The species of PAHs were analyzed, and the toxicity equivalency quantities (TEQs) of samples calculated. Analysis results indicate that the content of total PAHs in fly ash was 1.8 × 10{sup 3} times higher than that in bottom ash, and that the strongly carcinogenic PAHs with four or more rings accumulated sensitively in fly ash. The test results of samples gathered from open burning site demonstrate that Acenaphthylene (ACY), Acenaphthene (ACE), Fluorene (FLU), Phenanthrene (PHE), Anthracene (ANT) and other PAHs were inclined to migrate into surrounding environment along air and surface watershed corridors, while 4- to 6-ring PAHs accumulated more likely in soil. Being consistent with other studies, it has also been confirmed that increases in both free oxygen molecules and combustion temperatures could promote the decomposition of polycyclic PAHs. In addition, without the influence of combustion conditions, there is a positive correlation between total PCDD/Fs and total PAHs, although no such relationship has been found for TEQ.« less

Equine sperm-bound antisperm antibodies are associated with poor semen quality.

PubMed

Ferrer, M S; Miller, L M J

2018-06-01

Antisperm antibodies (ASAs) have been associated with infertility in stallions. The objectives of this study were to investigate the frequency of ASA-positive semen samples in satisfactory and non-satisfactory breeder stallions, the association between ASA binding and semen quality, and factors that may affect the diagnosis. Breeding soundness examinations were performed in 21 stallions and the percentage of IgG- and IgA-bound spermatozoa was evaluated using flow cytometry. Median IgG and IgA binding did not differ between the first and second ejaculates. The percentage of IgA-bound spermatozoa was higher in non-satisfactory (n = 10) than satisfactory breeder stallions (n = 11). However, IgG binding or frequency of IgG-positive ejaculates did not differ with stallion classification. The IgG-positive stallions had significantly lower total sperm motility, concentration and total numbers than IgG-negative stallions in the first ejaculate, and lower sperm concentration in the second ejaculate. The IgA-positive stallions had lower total sperm motility, normal spermatozoa and total numbers than IgA-negative stallions in the first ejaculate, and lower total sperm motility, normal spermatozoa and total numbers in the second ejaculate. While IgG binding did not differ with season, IgA binding was higher in the non-breeding season (n = 6 stallions) than the breeding season (n = 15 stallions) in the first ejaculate. Stallion age did not differ with ASA classification. In conclusion, IgG binding was highly prevalent in both groups of stallions, while IgA binding was higher and more prevalent in non-satisfactory breeders. Both isotypes were associated with poor semen quality. Season and sexual rest had an effect on IgA but not IgG binding. Copyright © 2018 Elsevier Inc. All rights reserved.

Male Partners' Investment and Alternatives as Correlates of Women's Coerced First Sexual Intercourse in Chinese Dating Relationships: A Social Exchange Perspective.

PubMed

He, Shanshan; Tsang, Sandra

2016-10-01

Social exchange theory has great potential to help our understanding of dating partners' sexual interaction. Yet, to our knowledge, there is still no empirical study applying this theory to explain sexual coercion in the context of intimate relationships. This study examined the relationship between male partners' social exchange variables (investment and alternatives) and women's coerced first sexual intercourse in dating relationships, within both gender samples (not dyadic data). A total of 927 valid questionnaires were collected by purposive snowball sampling in five main cities in China of college students who were currently in a romantic relationship. Results showed that in the male sample, male partners' investment significantly and positively correlated with emotional manipulation coercive tactics, and their alternatives significantly and positively predicted defection threats as coercive tactics. In the female sample, there were no such observations. The research hypotheses of this study have been partially supported, and its implications and limitations are discussed.

Antibiotic Residues in Milk from Three Popular Kenyan Milk Vending Machines.

PubMed

Kosgey, Amos; Shitandi, Anakalo; Marion, Jason W

2018-05-01

Milk vending machines (MVMs) are growing in popularity in Kenya and worldwide. Milk vending machines dispense varying quantities of locally sourced, pasteurized milk. The Kenya Dairy Board has a regulatory framework, but surveillance is weak because of several factors. Milk vending machines' milk is not routinely screened for antibiotics, thereby increasing potential for antibiotic misuse. To investigate, a total of 80 milk samples from four commercial providers ( N = 25), street vendors ( N = 21), and three MVMs ( N = 34) were collected and screened in Eldoret, Kenya. Antibiotic residue surveillance occurred during December 2016 and January 2017 using Idexx SNAP ® tests for tetracyclines, sulfamethazine, beta-lactams, and gentamicin. Overall, 24% of MVM samples and 24% of street vendor samples were presumably positive for at least one antibiotic. No commercial samples were positive. Research into cost-effective screening methods and increased monitoring by food safety agencies are needed to uphold hazard analysis and critical control point for improving antibiotic stewardship throughout the Kenyan private dairy industry.

Seroprevalence of Hepatitis A and E Virus Infections Among Healthy Population in Shiraz, Southern Iran.

PubMed

Asaei, Sadaf; Ziyaeyan, Mazyar; Moeini, Mahsa; Jamalidoust, Marzieh; Behzadi, Mohammad Amin

2015-07-01

Enterically-transmitted acute viral hepatitis is caused predominantly by hepatitis A virus (HAV) and hepatitis E virus (HEV). The prevalence of HEV and HAV infections varies in different geographical regions. This study was conducted to determine the prevalence of HEV and HAV infections among Iranian healthy individuals in southern Iran. Totally, 1030 samples were collected from healthy subjects in schools, those referred to tertiary outpatient clinics and health centers in Shiraz between November 2011 and May 2012. Their ages ranged between six months and 95 years. The presence of total anti-HAV and anti-HEV immunoglobulin M (IgM) in plasma was assessed by ELISA. The results showed that 66.2% and 0.6% of the general population in this area were positive for total anti-HAV and IgM antibodies by ELISA, respectively. As seen, 13.4% and 0.9% were positive for total anti-HEV and IgM antibodies, respectively. The difference in total anti-HAV and anti-HEV antibodies was significant among the age groups (P < 0.001). This study showed that the prevalence rates of HAV and HEV antibodies were positively correlated with age. The results demonstrated that the infection with these two viruses in the region was high and some high-risk individuals including females at child-bearing age were more susceptible. HAV vaccination could be recommended for antibody-negative adults.

Volume dependency for culture of fungi from respiratory secretions and increased sensitivity of Aspergillus quantitative PCR.

PubMed

Fraczek, Marcin G; Kirwan, Marie B; Moore, Caroline B; Morris, Julie; Denning, David W; Richardson, Malcolm D

2014-02-01

Diagnosis of aspergillosis is often difficult. We compared fungal yields from respiratory specimens using the Health Protection Agency standard culture method (BSOP57), a higher volume undiluted culture method Mycology Reference Centre Manchester (MRCM) and Aspergillus quantitative real time polymerase chain reaction (qPCR). Sputum, bronchial aspirate and bronchoalveolar lavage (BAL) samples (total 23) were collected from aspergillosis patients. One fraction of all samples was cultured using the MRCM method, one BSOP57 and one was used for qPCR. The recovery rate for fungi was significantly higher by MRCM (87%) than by BSOP57 (8.7%) from all 23 specimens. Sputum samples were 44% positive by MRCM compared to no fungi isolated (0%) by BSOP57. Bronchial aspirates were 75% positive by MRCM and 0% by BSOP57. BAL samples were positive in 20% by MRCM and 10% by BSOP57. qPCR was always more sensitive than culture (95.6%) from all samples. In general, over 100 mould colonies (81 Aspergillus fumigatus) were grown using the MRCM method compared with only one colony from BSOP57. This study provides a reference point for standardisation of respiratory sample processing in diagnostic laboratories. Culture from higher volume undiluted respiratory specimens has a much higher yield for Aspergillus than BSOP57. qPCR is much more sensitive than culture and the current UK method requires revision. © 2013 Blackwell Verlag GmbH.

Confirmation of congenital adrenal hyperplasia by adrenal steroid profiling of filter paper dried blood samples using ultra-performance liquid chromatography-tandem mass spectrometry.

PubMed

Rossi, Claudia; Calton, Lisa; Brown, Heather A; Gillingwater, Scott; Wallace, A Michael; Petrucci, Francesca; Ciavardelli, Domenico; Urbani, Andrea; Sacchetta, Paolo; Morris, Michael

2011-04-01

The specificity of screening for congenital adrenal hyperplasia by direct measurement of 17-hydroxyprogesterone in filter paper dried blood spot samples by immunoassay is low and has a high false-positive rate. In order to reduce the false-positive rate of this test, we developed a rapid, robust, specific confirmatory procedure in which cortisol, 4-androstene-3,17-dione and 17-hydroxyprogesterone were measured simultaneously by ultra-performance liquid chromatography-tandem mass spectrometry. After extraction, samples were analysed by ultra-performance liquid chromatography-tandem mass spectrometry and 17-hydroxyprogesterone was quantified accurately. Other steroids were determined using stable deuterated internal standards. In total, 25 patient blood spot samples and 92 control samples were analysed. The assay was linear for 17-hydroxyprogesterone, with a coefficient of determination >0.997 and imprecision ≤ 6.5%. An upper limit of normal for 17-hydroxyprogester-one of 4.45 nmol/L was established by analysing a cohort of samples from unaffected newborns. In addition, a cut-off of 3.5 for the peak areas ratio (17-hydroxyprogesterone+4-androstene-3,17-dione)/cortisol, allows confirmation of the affected steroidogenic enzyme. A high throughput method for the detection of steroids related to congenital adrenal hyperplasia has been developed, allowing the false-positive rate associated with screening for 17-hydroxyprogesterone by immunoassay to be determined.

Performance of the Fecal Immunochemical Test for Colorectal Cancer Screening Using Different Stool-Collection Devices: Preliminary Results from a Randomized Controlled Trial.

PubMed

Shin, Hye Young; Suh, Mina; Baik, Hyung Won; Choi, Kui Son; Park, Boyoung; Jun, Jae Kwan; Hwang, Sang-Hyun; Kim, Byung Chang; Lee, Chan Wha; Oh, Jae Hwan; Lee, You Kyoung; Han, Dong Soo; Lee, Do-Hoon

2016-11-15

We are in the process of conducting a randomized trial to determine whether compliance with the fecal immunochemical test (FIT) for colorectal cancer screening differs according to the stool-collection method. This study was an interim analysis of the performance of two stool-collection devices (sampling bottle vs conventional container). In total, 1,701 individuals (age range, 50 to 74 years) were randomized into the sampling bottle group (intervention arm) or the conventional container group (control arm). In both groups, we evaluated the FIT positivity rate, the positive predictive value for advanced neoplasia, and the detection rate for advanced neoplasia. The FIT positivity rates were 4.1% for the sampling bottles and 2.0% for the conventional containers; these values were significantly different. The positive predictive values for advanced neoplasia in the sampling bottles and conventional containers were 11.1% (95% confidence interval [CI], -3.4 to 25.6) and 12.0% (95% CI, -0.7 to 24.7), respectively. The detection rates for advanced neoplasia in the sampling bottles and conventional containers were 4.5 per 1,000 persons (95% CI, 2.0 to 11.0) and 2.4 per 1,000 persons (95% CI, 0.0 to 5.0), respectively. The impact of these findings on FIT screening performance was unclear in this interim analysis. This impact should therefore be evaluated in the final analysis following the final enrollment period.

Novel, improved sample preparation for rapid, direct identification from positive blood cultures using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry.

PubMed

Schubert, Sören; Weinert, Kirsten; Wagner, Chris; Gunzl, Beatrix; Wieser, Andreas; Maier, Thomas; Kostrzewa, Markus

2011-11-01

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is widely used for rapid and reliable identification of bacteria and yeast grown on agar plates. Moreover, MALDI-TOF MS also holds promise for bacterial identification from blood culture (BC) broths in hospital laboratories. The most important technical step for the identification of bacteria from positive BCs by MALDI-TOF MS is sample preparation to remove blood cells and host proteins. We present a method for novel, rapid sample preparation using differential lysis of blood cells. We demonstrate the efficacy and ease of use of this sample preparation and subsequent MALDI-TOF MS identification, applying it to a total of 500 aerobic and anaerobic BCs reported to be positive by a Bactec 9240 system. In 86.5% of all BCs, the microorganism species were correctly identified. Moreover, in 18/27 mixed cultures at least one isolate was correctly identified. A novel method that adjusts the score value for MALDI-TOF MS results is proposed, further improving the proportion of correctly identified samples. The results of the present study show that the MALDI-TOF MS-based method allows rapid (<20 minutes) bacterial identification directly from positive BCs and with high accuracy. Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

Detection of Cytomegalovirus (CMV) DNA in EDTA Whole-Blood Samples: Evaluation of the Quantitative artus CMV LightCycler PCR Kit in Conjunction with Automated Sample Preparation▿

PubMed Central

Michelin, Birgit D. A.; Hadžisejdić, Ita; Bozic, Michael; Grahovac, Maja; Hess, Markus; Grahovac, Blaženka; Marth, Egon; Kessler, Harald H.

2008-01-01

Whole blood has been found to be a reliable matrix for the detection and quantitation of cytomegalovirus (CMV) DNA. In this study, the performance of the artus CMV LightCycler (LC) PCR kit in conjunction with automated sample preparation on a BioRobot EZ1 workstation was evaluated. The accuracy, linearity, analytical sensitivity, and inter- and intra-assay variations were determined. A total of 102 clinical EDTA whole-blood samples were investigated, and results were compared with those obtained with the in vitro diagnostics (IVD)/Conformité Européene (CE)-labeled CMV HHV6,7,8 R-gene quantification kit. When the accuracy of the new kit was tested, seven of eight results were found to be within ±0.5 log10 unit of the expected panel results. Determination of linearity resulted in a quasilinear curve over more than 5 log units. The lower limit of detection of the assay was determined to be 139 copies/ml in EDTA whole blood. The interassay variation ranged from 15 to 58%, and the intra-assay variation ranged from 7 to 35%. When clinical samples were tested and the results were compared with those of the routinely used IVD/CE-labeled assay, 53 samples tested positive and 13 samples tested negative by both of the assays. One sample was found to be positive with the artus CMV LC PCR kit only, and 35 samples tested positive with the routinely used assay only. The majority of discrepant results were found with low-titer samples. In conclusion, use of the artus CMV LC PCR kit in conjunction with automated sample preparation on the BioRobot EZ1 workstation may be suitable for the detection and quantitation of CMV DNA in EDTA whole blood in the routine low-throughput laboratory; however, low-positive results may be missed by this assay. PMID:18272703

Detection of cytomegalovirus (CMV) DNA in EDTA whole-blood samples: evaluation of the quantitative artus CMV LightCycler PCR kit in conjunction with automated sample preparation.

PubMed

Michelin, Birgit D A; Hadzisejdic, Ita; Bozic, Michael; Grahovac, Maja; Hess, Markus; Grahovac, Blazenka; Marth, Egon; Kessler, Harald H

2008-04-01

Whole blood has been found to be a reliable matrix for the detection and quantitation of cytomegalovirus (CMV) DNA. In this study, the performance of the artus CMV LightCycler (LC) PCR kit in conjunction with automated sample preparation on a BioRobot EZ1 workstation was evaluated. The accuracy, linearity, analytical sensitivity, and inter- and intra-assay variations were determined. A total of 102 clinical EDTA whole-blood samples were investigated, and results were compared with those obtained with the in vitro diagnostics (IVD)/Conformité Européene (CE)-labeled CMV HHV6,7,8 R-gene quantification kit. When the accuracy of the new kit was tested, seven of eight results were found to be within +/-0.5 log(10) unit of the expected panel results. Determination of linearity resulted in a quasilinear curve over more than 5 log units. The lower limit of detection of the assay was determined to be 139 copies/ml in EDTA whole blood. The interassay variation ranged from 15 to 58%, and the intra-assay variation ranged from 7 to 35%. When clinical samples were tested and the results were compared with those of the routinely used IVD/CE-labeled assay, 53 samples tested positive and 13 samples tested negative by both of the assays. One sample was found to be positive with the artus CMV LC PCR kit only, and 35 samples tested positive with the routinely used assay only. The majority of discrepant results were found with low-titer samples. In conclusion, use of the artus CMV LC PCR kit in conjunction with automated sample preparation on the BioRobot EZ1 workstation may be suitable for the detection and quantitation of CMV DNA in EDTA whole blood in the routine low-throughput laboratory; however, low-positive results may be missed by this assay.

The occurrence and significance to animal health of salmonellas in sewage and sewage sludges.

PubMed Central

Jones, P. W.; Rennison, L. M.; Lewin, V. H.; Redhead, D. L.

1980-01-01

A total of 882 samples of settled sewage, sewage sludges and final effluents from eight sewage treatment plants were examined for the presence of salmonellas. Of these samples 68% were positive, isolations being made most frequently from settled sewage (85%), raw sludge (87%) and anaerobically digested sludge (96%). Fewer isolations were made from final effluent (24%) and processed sludges (58%). Samples usually contained less than 200 salmonellas/100 ml and arguments are presented that such concentrations should not lead to disease in animals if suitable grazing restrictions are followed. PMID:6985928

Residual pyrethroids in fresh horticultural products in Sonora, Mexico.

PubMed

Aldana-Madrid, Maria L; Valenzuela-Quintanar, Ana I; Silveira-Gramont, Maria I; Rodríguez-Olibarría, Guillermo; Grajeda-Cota, Patricia; Zuno-Floriano, Fabiola G; Miller, Marion G

2011-10-01

This study was conducted to evaluate the presence of cyhialothrin, cyfluthrin, cypermethrin, fenvalerate, and deltamethrin in vegetables produced and consumed in Sonora, Mexico. A total of 345 samples were collected from cluster sampling of markets and fields. Approximately 9% of the samples tested positive for pyrethroids (residue range 0.004-0.573 mg kg(-1)). Based on the results, the potential toxicological risk of human exposure to the pyrethroid insecticides measured in vegetables appears to be minimal, with the estimated exposure being 1,000 times lower than admissible levels. © Springer Science+Business Media, LLC 2011

Lower urinary tract infection and bacterial colonization in patient with double J ureteral stent.

PubMed

Joshi, R; Singh, D R; Sharma, S

2011-10-01

The aim of this study is to investigate the bacteriology of urinary tract infection associated with indwelling DJ stent. A total of 60 patients were included and 14 lost during follow up. Study period was for 6 months carried out in the department of surgery, Kathmandu Medical College. Prophylactic antibiotic was given at the time of intervention. Mid stream urine samples for routine and culture were sent before intervention. Urine samples during DJ removal and DJ tip cultures were also sent. All patients were "stented" during the various open and endourolgical procedures. Patients were clinically followed for a period till the DJ was removed. Statistical Package for Scientific Study (SPSS) 11, Chi square Test was used for statistical analysis. A total of 46 cases were included. Mean age in years was 35.70 (10-78 years). Male were 22 and female 24. Eleven patients (23.91%) had stent placed less than 30 days and 35 patients (76.08%) had it for equal or more than 30 days. DJ indwelling time was in between 12-86 days. Bacterial colonies were found in 28.3% (13 of 46) of the urine samples and 30.4% (14 of 46) from the tip of the DJ stent segment. Of the pathogens identified, E. coli was found to be the most common. An increased stent colonization rate was associated with implantation time, female sex. On urine culture 70.21% had no growth, 14.89% E. coli, 4.25% Klebsiella, Actinobacter, 2.12% E. coli/kleb, multiple org, psuedomonas. Ten patients (21.7%) had positive urine culture before stent insertion. Thirteen patients (28.3%) were shown to have positive urine culture on stent removal. Fourteen patients (30.4%) had positive DJ stent culture. Positive urine culture and positive DJ tip cultures had strong correlation. Longer duration of placement of stent showed stent colonization. The commonest pathogen was E. coli.

Characterization of West Nile Viruses Isolated from Captive American Flamingoes (Phoenicopterus ruber) in Medellin, Colombia

PubMed Central

Osorio, Jorge E.; Ciuoderis, Karl A.; Lopera, Juan G.; Piedrahita, Leidy D.; Murphy, Darby; LeVasseur, James; Carrillo, Lina; Ocampo, Martha C.; Hofmeister, Erik

2012-01-01

Serum samples from a total of 71 healthy captive birds belonging to 18 species were collected in July of 2008 in Medellin (Colombia) and tested for flaviviruses. Eighteen of 29 samples from American Flamingoes (Phoenicopterus ruber) were positive for West Nile virus (WNV) by reverse transcription-polymerase chain reaction. Selected positive samples were serially passaged and WNV was confirmed by immunofluorescence. Two isolates (524/08, 9835/08) were characterized in vitro and in vivo. Sequence analysis revealed WNV with 16 nucleotide substitutions resulting in six amino acid changes when compared with the NY99 strain. Colombian (COL) viruses were more closely related to Louisiana isolates from 2001. When compared with attenuated strains isolated from Texas, COL isolates differed in their plaque size and temperature sensitivity phenotype. The COL viruses were pathogenic in embryonated chicken eggs and Balb/c mice. PMID:22802436

Prevalence and Molecular Analyses of Hemotrophic Mycoplasma spp. (Hemoplasmas) Detected in Sika Deer (Cervus nippon yesoensis) in Japan

PubMed Central

TAGAWA, Michihito; MATSUMOTO, Kotaro; YOKOYAMA, Naoaki; INOKUMA, Hisashi

2013-01-01

ABSTRACT Hemotropic mycoplasmas (hemoplasmas) are cell-wall deficient, epierythrocytic bacteria that cause infectious anemia in several mammalian species. The prevalence of hemoplasma species was examined by screening and species-specific PCR using blood samples collected from 51 sika deer in Hokkaido, Japan. Molecular analyses were performed for the 16S rRNA, 23S rRNA and RNase P RNA (rnpB) gene sequences. A total of 23/51 (45%) deer DNA samples were positive for hemoplasmas in the screening PCR. Using species-specific PCR, 12 and 17 samples were positive for ‘Candidatus Mycoplasma haemocervae’ and ‘Candidatus M. erythrocervae’, respectively. Sequencing and phylogenetic trees of those three genes indicate that the ‘Candidatus M. haemocervae’ and ‘Candidatus M. erythrocervae’ detected in Japanese deer are potentially different species from the cervine hemoplasma found in deer from America and Brazil. PMID:24270803

Serologic evidence of exposure of raptors to influenza A virus.

PubMed

Redig, Patrick T; Goyal, Sagar M

2012-06-01

Serum or plasma samples from raptors that prey or scavenge upon aquatic birds were tested by a commercially available blocking enzyme-linked immunosorbent assay for the evidence of antibodies to influenza A virus. Samples were taken from birds (n = 616) admitted to two rehabilitation centers in the United States. In addition, samples from 472 migrating peregrine falcons (Falco peregrinus) trapped on autumnal and vernal migrations for banding purposes were also tested. Only bald eagles were notably seropositive (22/406). One each of peregrine falcon, great horned owl (Bubo virginianus), and Cooper's hawk (Accipiter cooperi) from a total of 472, 81, and 100, respectively, were also positive. None of the turkey vultures (n = 21) or black vultures (n = 8) was positive. No clinical signs referable to avian influenza were seen in any bird at the time of capture. These data indicate that, among raptors, bald eagles do have exposure to influenza A viruses.

Characterization of West Nile viruses isolated form captive American flamingoes (Phoenicopterus ruber) in Medellin, Colombia.

USGS Publications Warehouse

Osorio, Jorge E.; Ciuoderis, Karl A.; Lopera, Juan G.; Piedrahita, Leidy D.; Murphy, Darby; LeVasseur, James; Carrillo, Lina; Ocampo, Martha C.; Hofmeister, Erik

2012-01-01

Serum samples from a total of 71 healthy captive birds belonging to 18 species were collected in July of 2008 in Medellin (Colombia) and tested for flaviviruses. Eighteen of 29 samples from American Flamingoes (Phoenicopterus ruber) were positive for West Nile virus (WNV) by reverse transcription-polymerase chain reaction. Selected positive samples were serially passaged and WNV was confirmed by immunofluorescence. Two isolates (524/08, 9835/08) were characterized in vitro and in vivo. Sequence analysis revealed WNV with 16 nucleotide substitutions resulting in six amino acid changes when compared with the NY99 strain. Colombian (COL) viruses were more closely related to Louisiana isolates from 2001. When compared with attenuated strains isolated from Texas, COL isolates differed in their plaque size and temperature sensitivity phenotype. The COL viruses were pathogenic in embryonated chicken eggs and Balb/c mice.

Molecular Typing of Treponema pallidum in the Czech Republic during 2011 to 2013: Increased Prevalence of Identified Genotypes and of Isolates with Macrolide Resistance

PubMed Central

Grillová, Linda; Pĕtrošová, Helena; Mikalová, Lenka; Strnadel, Radim; Dastychová, Eliška; Kuklová, Ivana; Kojanová, Martina; Kreidlová, Miluše; Vaňousová, Daniela; Hercogová, Jana; Procházka, Přemysl; Zákoucká, Hana; Krchňáková, Alena; Vašků, Vladimír

2014-01-01

From January 2011 to December 2013, a total of 262 samples, from 188 patients suspected of having syphilis were tested for the presence of treponemal DNA by PCR amplification of five chromosomal loci, including the polA (TP0105), tmpC (TP0319), TP0136, TP0548, and 23S rRNA genes. Altogether, 146 samples from 103 patients were PCR positive for treponemal DNA. A set of 81 samples from 62 PCR-positive patients were typeable, and among them, nine different genotypes were identified. Compared to a previous study in the Czech Republic during 2004 to 2010, the number of genotypes detected among syphilis patients in a particular year increased to six in both 2012 and 2013, although they were not the same six. The proportion of macrolide-resistant clinical isolates in this 3-year study was 66.7%. PMID:25100820

PERFORMANCE EVALUATION OF A PROTOTYPE ARCHITECT ANTIBODY ASSAY FOR BABESIA MICROTI.

PubMed

Cheng, Kevin; Coller, Kelly E; Marohnic, Christopher C; Pfeiffer, Zachary A; Fino, James R; Elsing, Randee R; Bergsma, Janet; Marcinkus, Marilee A; Kar, Alak K; Gumbs, Orlando H; Otis, Kathy S; Fishpaugh, Jeffrey; Schultz, Phillip W; Pope, Mark R; Narvaez, Alfredo R; Wong, Susan J; Madison-Antenucci, Susan; Leary, Thomas P; Dawson, George J

2018-05-09

The tick-borne protozoan Babesia microti is responsible for more than 200 cases of transfusion-transmitted babesiosis (TTB) infection in the United States over the last 30 years. Measures to mitigate the risk of TTB include nucleic acid testing (NAT) and B. microti antibody testing. A fully automated prototype B. microti antibody test was developed on the ARCHITECT instrument. The specificity was determined to be 99.98% in volunteer blood donors (n=28,740) from areas considered as low endemic for B. microti The sensitivity of the prototype test was studied in experimentally-infected macaques; a total of 128 samples were detected compared to 125 with the indirect fluorescent antibody test (IFA), additionally, 83 (89.2%) of the PCR positive samples were detected compared to 81 (87.1%) using the IFA test. All PCR positive samples that tested negative in the prototype antibody test were pre-seroconversion period samples. Following seroconversion, periods of intermittent parasitemia occurred; 17 PCR negative samples drawn in-between PCR positive bleed dates, tested positive both by the prototype test (robust reactivity) and IFA (marginal reactivity) prior to the administration of therapeutic drugs, indicating that the PCR test failed to detect samples from persistently infected macaques. The prototype assay detected 56 of 58 (96.6%) human subjects diagnosed with clinical babesiosis by both PCR and IFA testing. Overall, the prototype anti-babesia assay provides a highly sensitive and specific test for the diagnosis of B. microti infection. While PCR is preferred for detection of window period parasitemia, antibody tests detect infected subjects during periods of low level parasitemia. Copyright © 2018 Cheng et al.

The zoonotic potential of Giardia intestinalis assemblage E in rural settings.

PubMed

Abdel-Moein, Khaled A; Saeed, Hossam

2016-08-01

Giardiasis is a globally re-emerging protozoan disease with veterinary and public health implications. The current study was carried out to investigate the zoonotic potential of livestock-specific assemblage E in rural settings. For this purpose, a total of 40 microscopically positive Giardia stool samples from children with gastrointestinal complaints with or without diarrhea were enrolled in the study as well as fecal samples from 46 diarrheic cattle (18 dairy cows and 28 calves). Animal samples were examined by sedimentation method to identify Giardia spp., and then, all Giardia positive samples from human and animals were processed for molecular detection of livestock-specific assemblage E through amplification of assemblage-specific triosephosphate isomerase (tpi) gene using nested polymerase chain reaction (PCR). The results of the study revealed high unexpected occurrence of assemblage E among human samples (62.5 %), whereas the distribution among patients with diarrhea and those without was 42.1 and 81 %, respectively. On the other hand, the prevalence of Giardia spp. among diarrheic dairy cattle was (8.7 %), while only calves yielded positive results (14.3 %) and all bovine Giardia spp. were genetically classified as Giardia intestinalis assemblage E. Moreover, DNA sequencing of randomly selected one positive human sample and another bovine one revealed 100 and 99 % identity with assemblage E tpi gene sequences available at GenBank after BLAST analysis. In conclusion, the current study highlights the wide dissemination of livestock-specific assemblage E among humans in rural areas, and thus, zoonotic transmission cycle should not be discounted during the control of giardiasis in such settings.

Molecular characterization of chicken infectious anemia virus from contaminated live-virus vaccines.

PubMed

Li, Yang; Hu, Yan; Cui, Shuai; Fu, Jiayuan; Wang, Yixin; Cui, Zhizhong; Fang, Lichun; Chang, Shuang; Zhao, Peng

2017-05-01

The aim of this study was to investigate possible causes of the pervasiveness of chicken infectious anemia virus (CIAV) infection in chickens in recent years in China. A total of 14 batches of live-virus vaccines were examined using PCR to detect CIAV contamination, of which only 2 samples (a Newcastle disease vaccine and a fowl pox vaccine) tested positive for CIAV. These Newcastle and fowl pox vaccines were then inoculated into 1-day-old specific-pathogen-free chickens. Serum samples were collected from chickens infected with the PCR-positive vaccines, and these tested positive for CIAV-specific antibodies as tested using ELISA. In addition, DNA samples isolated from the serum samples also tested positive by PCR. The results indicated that the samples were contaminated with CIAV and identified 2 exogenous CIAV strains, designated CIAV-N22 and CIAV-F10, in the respective samples. The full genome sequences of these novel CIAV strains were sequenced and analyzed. Phylogenetic tree analysis indicated that the CIAV-F10 strain might represent a recombinant viral strain arising from the parental CIAV strains JQ690762 and KJ728816. Overall, the results suggested that vaccination with CIAV-contaminated vaccines contributed to the prevalence and spread of CIAV infection in chickens. Furthermore, the CIAV contaminant was likely subsequently transmitted to commercial chickens through congenital transmission. Our findings therefore highlight the need for more extensive screening of live-virus vaccines for poultry in China to reduce the threat of contamination with exogenous viruses. © 2016 Poultry Science Association Inc.

Assessing total nitrogen in surface-water samples--precision and bias of analytical and computational methods

USGS Publications Warehouse

Rus, David L.; Patton, Charles J.; Mueller, David K.; Crawford, Charles G.

2013-01-01

The characterization of total-nitrogen (TN) concentrations is an important component of many surface-water-quality programs. However, three widely used methods for the determination of total nitrogen—(1) derived from the alkaline-persulfate digestion of whole-water samples (TN-A); (2) calculated as the sum of total Kjeldahl nitrogen and dissolved nitrate plus nitrite (TN-K); and (3) calculated as the sum of dissolved nitrogen and particulate nitrogen (TN-C)—all include inherent limitations. A digestion process is intended to convert multiple species of nitrogen that are present in the sample into one measureable species, but this process may introduce bias. TN-A results can be negatively biased in the presence of suspended sediment, and TN-K data can be positively biased in the presence of elevated nitrate because some nitrate is reduced to ammonia and is therefore counted twice in the computation of total nitrogen. Furthermore, TN-C may not be subject to bias but is comparatively imprecise. In this study, the effects of suspended-sediment and nitrate concentrations on the performance of these TN methods were assessed using synthetic samples developed in a laboratory as well as a series of stream samples. A 2007 laboratory experiment measured TN-A and TN-K in nutrient-fortified solutions that had been mixed with varying amounts of sediment-reference materials. This experiment identified a connection between suspended sediment and negative bias in TN-A and detected positive bias in TN-K in the presence of elevated nitrate. A 2009–10 synoptic-field study used samples from 77 stream-sampling sites to confirm that these biases were present in the field samples and evaluated the precision and bias of TN methods. The precision of TN-C and TN-K depended on the precision and relative amounts of the TN-component species used in their respective TN computations. Particulate nitrogen had an average variability (as determined by the relative standard deviation) of 13 percent. However, because particulate nitrogen constituted only 14 percent, on average, of TN-C, the precision of the TN-C method approached that of the method for dissolved nitrogen (2.3 percent). On the other hand, total Kjeldahl nitrogen (having a variability of 7.6 percent) constituted an average of 40 percent of TN-K, suggesting that the reduced precision of the Kjeldahl digestion may affect precision of the TN-K estimates. For most samples, the precision of TN computed as TN-C would be better (lower variability) than the precision of TN-K. In general, TN-A precision (having a variability of 2.1 percent) was superior to TN-C and TN-K methods. The laboratory experiment indicated that negative bias in TN-A was present across the entire range of sediment concentration and increased as sediment concentration increased. This suggested that reagent limitation was not the predominant cause of observed bias in TN-A. Furthermore, analyses of particulate nitrogen present in digest residues provided an almost complete accounting for the nitrogen that was underestimated by alkaline-persulfate digestion. This experiment established that, for the reference materials at least, negative bias in TN-A was caused primarily by the sequestration of some particulate nitrogen that was refractory to the digestion process. TN-K biases varied between positive and negative values in the laboratory experiment. Positive bias in TN-K is likely the result of the unintended reduction of a small and variable amount of nitrate to ammonia during the Kjeldahl digestion process. Negative TN-K bias may be the result of the sequestration of a portion of particulate nitrogen during the digestion process. Negative bias in TN-A was present across the entire range of suspended-sediment concentration (1 to 14,700 milligrams per liter [mg/L]) in the synoptic-field study, with relative bias being nearly as great at sediment concentrations below 10 mg/L (median of -3.5 percent) as that observed at sediment concentrations up to 750 mg/L (median of -4.4 percent). This lent support to the laboratory-experiment finding that some particulate nitrogen is sequestered during the digestion process, and demonstrated that negative TN-A bias was present in samples with very low suspended-sediment concentrations. At sediment concentrations above 750 mg/L, the negative TN-A bias became more likely and larger (median of -13.2 percent), suggesting a secondary mechanism of bias, such as reagent limitation. From a geospatial perspective, trends in TN-A bias were not explained by selected basin characteristics. Though variable, TN-K bias generally was positive in the synoptic-field study (median of 3.1 percent), probably as a result of the reduction of nitrate. Three alternative approaches for assessing TN in surface water were evaluated for their impacts on existing and future sampling programs. Replacing TN-A with TN-C would remove the bias from subsequent data, but this approach also would introduce discontinuity in historical records. Replacing TN-K with TN-C would lead to the removal of positive bias in TN-K in the presence of elevated nitrate. However, in addition to the issues that may arise from a discontinuity in the data record, this approach may not be applicable to regulatory programs that require the use of total Kjeldahl nitrogen for stream assessment. By adding TN-C to existing TN-A or TN-K analyses, historical-data continuity would be preserved and the transitional period could be used to minimize the impact of bias on data analyses. This approach, however, imposes the greatest burdens on field operations and in terms of analytical costs. The variation in these impacts on different sampling programs will challenge U.S. Geological Survey scientists attempting to establish uniform standards for TN sample collection and analytical determinations.

Expanding STEM opportunities through inclusive STEM‐focused high schools

PubMed Central

Wang, Haiwen; Wei, Xin; Lynch, Sharon; Peters, Vanessa; Young, Viki; Allen, Carrie

2017-01-01

Abstract Inclusive STEM high schools (ISHSs) (where STEM is science, technology, engineering, and mathematics) admit students on the basis of interest rather than competitive examination. This study examines the central assumption behind these schools—that they provide students from subgroups underrepresented in STEM with experiences that equip them academically and attitudinally to enter and stay in the STEM pipeline. Hierarchical modeling was applied to data from student surveys and state longitudinal data records for 5113 students graduating from 39 ISHSs and 22 comprehensive high schools in North Carolina and Texas. Compared to peers from the same demographic group with similar Grade 8 achievement levels, underrepresented minority and female ISHS students in both states were more likely to undertake advanced STEM coursework. Hispanics in Texas and females in both states expressed more STEM career interest in Grade 12 if they attended an ISHS. Positive relationships between ISHS attendance and grade point average were found in the total sample and each subgroup in North Carolina. Positive ISHS advantages in terms of test scores for the total student sample were found for science in both states and for mathematics in Texas. For the various student subgroups, test score differences favored the ISHS samples but attained statistical significance only for African Americans’ science achievement scores in the Texas study. PMID:28919649

Fast preparation of a long chimeric armored RNA as controls for external quality assessment for molecular detection of Zika virus.

PubMed

Lin, Guigao; Zhang, Kuo; Zhang, Dong; Han, Yanxi; Xie, Jiehong; Li, Jinming

2017-03-01

The emergence of Zika virus demands accurate laboratory diagnostics. Nucleic acid testing is currently the definitive method for diagnosis of Zika infection. In 2016, an external quality assurance (EQA) for assessing the quality of molecular testing of Zika virus was carried out in China. A single armored RNA encapsulating a 4942-nucleotides (nt) long specific RNA sequence of Zika virus was prepared and used as positive samples. A pre-tested EQA panel, consisting of 4 negative and 6 positive samples with different concentrations of armored RNA, was distributed to 38 laboratories that perform molecular detection of Zika virus. A total of 39 data sets (1 laboratory used two test kits in parallel), produced by using commercial (n=38) or laboratory developed (n=1) quantitative reverse-transcriptase PCR (qRT-PCR) kits, were received. Of these, 35 (89.7%) had correct results for all 10 samples, and 4 (10.3%) reported at least 1 error (11 in total). The testing errors were all false-negatives, highlighting the need of improvements in detecting sensitivity. The EQA reveals that the majority of participating laboratories are proficient in molecular testing of Zika virus. Copyright © 2017 Elsevier B.V. All rights reserved.

Occurrence of gastrointestinal parasites in wild animals in State of Paraná, Brazil.

PubMed

Sprenger, Lew K; Yoshitani, Ursula Y; Buzatti, Andreia; Molento, Marcelo B

2018-01-01

The objective of this study was to determine the prevalence and diversity of gastrointestinal parasites in fecal samples from wild birds and mammals from the State of Paraná. In total, 220 stool samples were sent to Parasitic Diseases Laboratory of the Federal University of Paraná during 13 months (Jan/2013-Jan/2014). A total of 52.7% (116/220) of the animals were positive for cysts, oocysts, eggs and/or trophozoites. In birds, the positivity rate was 37.9% (25/66) and mammals was 59.1% (91/154). Strongyloidea superfamily eggs were observed in 37.3% (82/220) of the samples, Eimeria spp. in 10% (22/220), and Trichuris spp. in 4.5% (10/220). The most frequent mammal species were llamas (Lama glama), and dromedaries (Camelus bactrianus) with infection rate of 70.1% (54/77) and 60.8% (14/23), respectively. In other hand, cockatiels (Nymphicus hollandicus) and ring necks (Psittacula krameri), were the most researched birds, with infection rate of 20% (40/50) and 100% (6/6), respectively. A high prevalence of gastrointestinal parasites was observed in most of wildlife animals. Further investigations should be conducted focusing on parasite control strategies and the conservation measurements for harmonizing the human-animal interaction on the long-term, reducing associated health risks.

Exploring chemical variables in Ligustrum lucidum Ait. F. tricolor (rehd.) Rehd. in relation to air pollutants and environmental conditions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pignata, M.L.; Canas, M.S.; Carreras, H.A.

1997-09-01

A diagnostic study was done on Ligustrum lucidum Ait. f. tricolor (Rehd.) Rehd. in relation to atmospheric pollutants in Cordoba city, Argentina. The study area receives regional Pollutants and was categorized taking into account traffic level, industrial density, type of industry, location of the sample point in relation to the street corner, treeless condition, and topographic level. Dried weight/fresh weight ratio (DW/FW) and specific leaf area (SLA) were calculated, and concentrations of chlorophylls, carotenoids, total sulfur, soluble proteins, malondialdehyde (MDA), and hydroperoxy conjugated dienes (HPCD) were determined in leaf samples. Sulfur content correlates positively with traffic density and SLA correlatesmore » negatively with some combinations of the categorical variables; MDA correlates positively with topographic level and total protein concentration correlates negatively with treeless condition. On the basis of our results, traffic, location of trees, type of industry, situation of a tree with respect to others, and topographic level are the environmental variables to bear in mind when selecting analogous sampling points in a passive monitoring program. An approximation to predict tree injury may be obtained by measuring DW/FW ratio, proteins, pigments, HPCD, and MDA as they are responsible for the major variability of data.« less

Expanding STEM opportunities through inclusive STEM-focused high schools.

PubMed

Means, Barbara; Wang, Haiwen; Wei, Xin; Lynch, Sharon; Peters, Vanessa; Young, Viki; Allen, Carrie

2017-09-01

Inclusive STEM high schools (ISHSs) (where STEM is science, technology, engineering, and mathematics) admit students on the basis of interest rather than competitive examination. This study examines the central assumption behind these schools-that they provide students from subgroups underrepresented in STEM with experiences that equip them academically and attitudinally to enter and stay in the STEM pipeline. Hierarchical modeling was applied to data from student surveys and state longitudinal data records for 5113 students graduating from 39 ISHSs and 22 comprehensive high schools in North Carolina and Texas. Compared to peers from the same demographic group with similar Grade 8 achievement levels, underrepresented minority and female ISHS students in both states were more likely to undertake advanced STEM coursework. Hispanics in Texas and females in both states expressed more STEM career interest in Grade 12 if they attended an ISHS. Positive relationships between ISHS attendance and grade point average were found in the total sample and each subgroup in North Carolina. Positive ISHS advantages in terms of test scores for the total student sample were found for science in both states and for mathematics in Texas. For the various student subgroups, test score differences favored the ISHS samples but attained statistical significance only for African Americans' science achievement scores in the Texas study.

Sensitivity of bhk-21 cells supplemented with diethylaminoethyl-dextran for detection of street rabies virus in saliva samples.

PubMed Central

Larghi, O P; Nebel, A E; Lazaro, L; Savy, V L

1975-01-01

A tissue culture system for detecting rabies virus from saliva samples of suspected animals was developed and compared to suckling mouse inoculation. Swab samples were obtained from the mouth of the animal heads received for rabies diagnosis; these swabs were submerged in maintenance medium. The maintenance medium was inoculated intracerebrally into suckling mice and onto BHK-21 cells with diethylaminoethyl (DEAE)-dextran (BHK/DEAE) and without (BHK). Rabies immunofluorescence was performed on the brain of the mice dying during the observation period and also on both tissue culture systems every day after infection. The BHK-DEAE system detected 28 positive samples obtained from 48 rabid animals and the BHK system detected 18. By suckling mouse inoculation only 11 of the same positive samples were detected. A total of 90 samples was studied by the three methods. Rabies virus was detected by the tissue culture methods earlier than by suckling mouse inoculation. The BHK-DEAE method was an economic and fast method for rabies virus detection in saliva samples, which could be used for ecological and pathogenesis studies, as well for rabies diagnosis before the death of the suspected animal. PMID:1100655

Predictors of Multiple Suicide Attempts among Suicidal Black Adolescents

PubMed Central

Merchant, Christopher; Kramer, Anne; Joe, Sean; Venkataraman, Sanjeev; King, Cheryl A.

2015-01-01

Psychopathology, social support, and interpersonal orientation were studied in relation to suicide attempt status in acutely suicidal, psychiatrically hospitalized Black adolescents and a matched sample of White adolescents. In the total sample, multiple attempters were differentiated by lower perceived support. Within the Black youth subsample, social comparison and positive stimulation from others differentiated multiple attempters from single attempters/ideators. Only suicidal ideation predicted multiple attempts among White youth and only higher interpersonal orientation predicted multiple suicide attempts within Black adolescents. PMID:19527152

Predictors of multiple suicide attempts among suicidal black adolescents.

PubMed

Merchant, Christopher; Kramer, Anne; Joe, Sean; Venkataraman, Sanjeev; King, Cheryl A

2009-04-01

Psychopathology, social support, and interpersonal orientation were studied in relation to suicide attempt status in acutely suicidal, psychiatrically hospitalized Black adolescents and a matched sample of White adolescents. In the total sample, multiple attempters were differentiated by lower perceived support. Within the Black youth subsample, social comparison and positive stimulation from others differentiated multiple attempters from single attempters/ideators. Only suicidal ideation predicted multiple attempts among White youth and only higher interpersonal orientation predicted multiple suicide attempts within Black adolescents.

Assessment of region, farming system, irrigation source and sampling time as food safety risk factors for tomatoes.

PubMed

Pagadala, Sivaranjani; Marine, Sasha C; Micallef, Shirley A; Wang, Fei; Pahl, Donna M; Melendez, Meredith V; Kline, Wesley L; Oni, Ruth A; Walsh, Christopher S; Everts, Kathryne L; Buchanan, Robert L

2015-03-02

In the mid-Atlantic region of the United States, small- and medium-sized farmers use varied farm management methods and water sources to produce tomatoes. It is unclear whether these practices affect the food safety risk for tomatoes. This study was conducted to determine the prevalence, and assess risk factors for Salmonella enterica, Shiga toxin-producing Escherichia coli (STEC) and bacterial indicators in pre-harvest tomatoes and their production areas. A total of 24 organic and conventional, small- to medium-sized farms were sampled for six weeks in Maryland (MD), Delaware (DE) and New Jersey (NJ) between July and September 2012, and analyzed for indicator bacteria, Salmonella and STEC. A total of 422 samples--tomato fruit, irrigation water, compost, field soil and pond sediment samples--were collected, 259 of which were tomato samples. A low level of Salmonella-specific invA and Shiga toxin genes (stx1 or stx2) were detected, but no Salmonella or STEC isolates were recovered. Of the 422 samples analyzed, 9.5% were positive for generic E. coli, found in 5.4% (n=259) of tomato fruits, 22.5% (n=102) of irrigation water, 8.9% (n=45) of soil, 3/9 of pond sediment and 0/7 of compost samples. For tomato fruit, farming system (organic versus conventional) was not a significant factor for levels of indicator bacteria. However, the total number of organic tomato samples positive for generic E. coli (1.6%; 2/129) was significantly lower than for conventional tomatoes (6.9% (9/130); (χ(2) (1)=4.60, p=0.032)). Region was a significant factor for levels of Total Coliforms (TC) (p=0.046), although differences were marginal, with western MD having the highest TC counts (2.6 log CFU/g) and NJ having the lowest (2.0 log CFU/g). Tomatoes touching the ground or plastic mulch harbored significantly higher levels of TC compared to vine tomatoes, signaling a potential risk factor. Source of irrigation water was a significant factor for all indicator bacteria (p<0.0001), and groundwater had lower bacterial levels than surface water. End of line surface water samples were not significantly different from source water samples, but end of line groundwater samples had significantly higher bacterial counts than source (p<0.0001), suggesting that Good Agricultural Practices that focus on irrigation line maintenance might be beneficial. In general, local effects other than cropping practices, including topography, land use and adjacent industries, might be important factors contributing to microbiological inputs on small- and medium-sized farms in the mid-Atlantic region. Copyright © 2014 Elsevier B.V. All rights reserved.

Comparison of Subjective Wellbeing and Positive Future Expectations in Between Working and Nonworking Adolescents in Turkey.

PubMed

Karaca, Semra; Karakoc, Ayse; Bingol, Fadime; Eren, Nurhan; Andsoy, Isil Isik

2016-02-01

Wellbeing is one's evaluation and judgment of one's life. It consists of 3 dimensions: positive affectivity, negative affectivity, and life satisfaction. This study aimed to compare the subjective wellbeing and positive future expectations between working and nonworking adolescents. The study was designed as descriptive and comparative. The study sample consisted of 420 working and 482 nonworking adolescents (n = 902) aged 15 - 20 years, who were randomly recruited from two occupational education centers in Istanbul, Turkey and two high schools (formal educations) in the same district. Adolescent subjective wellbeing scale (ASWS) total mean (SD) scores for working adolescents and nonworking adolescents were 48.76 (9.50) and 49.72 (8.01), respectively. In addition, positive future expectations scale (PFES) total mean (SD) scores for working adolescents and nonworking adolescents were 18.71 (4.50) and 19.06 (3.49), respectively. In this study, no significant difference was found between the general wellbeing (scale total median score) scores of the working and nonworking adolescent groups (Z = 1.01, P = 0.315). However, significant differences were found in the family relations satisfaction (Z = 3.23, P = 0.002) and relations with significant others (Z = 2.85, P = 0.004) subscales of the ASWS. A positive relationship was found between adolescent subjective wellbeing and positive future expectations. It was found that nonworking adolescents scored higher on the dimensions of "family relations" and "relations with significant others" of subjective wellbeing compared to those dimensions in working adolescents.

Evaluative Conditioning in Humans: A Meta-Analysis

ERIC Educational Resources Information Center

Hofmann, Wilhelm; De Houwer, Jan; Perugini, Marco; Baeyens, Frank; Crombez, Geert

2010-01-01

This article presents a meta-analysis of research on "evaluative conditioning" (EC), defined as a change in the liking of a stimulus (conditioned stimulus; CS) that results from pairing that stimulus with other positive or negative stimuli (unconditioned stimulus; US). Across a total of 214 studies included in the main sample, the mean…

Drinking and Driving PSAs: A Content Analysis of Behavioral Influence Strategies.

ERIC Educational Resources Information Center

Slater, Michael D.

1999-01-01

Study randomly samples 66 drinking and driving television public service announcements that were then coded using a categorical and dimensional scheme. Data set reveals that informational/testimonial messages made up almost half of the total; positive appeals were the next most common, followed by empathy, fear, and modeling appeals. (Contains 34…

SOURCE APPORTIONMENT OF PM2.5 AT AN URBAN IMPROVE SITE IN SEATTLE, WA

EPA Science Inventory

The multivariate receptor models Positive Matrix Factorization (PMF) and Unmix were used along with EPA's Chemical Mass Balance model to deduce the sources of PM2.5 at a centrally located urban site in Seattle, Washington. A total of 289 filter samples were obtained with an IM...

40 CFR 141.860 - Violations.

Code of Federal Regulations, 2014 CFR

2014-07-01

... PRIMARY DRINKING WATER REGULATIONS Revised Total Coliform Rule § 141.860 Violations. (a) E. coli MCL Violation. A system is in violation of the MCL for E. coli when any of the conditions identified in paragraphs (a)(1) through (a)(4) of this section occur. (1) The system has an E. coli-positive repeat sample...

40 CFR 141.860 - Violations.

Code of Federal Regulations, 2013 CFR

2013-07-01

... PRIMARY DRINKING WATER REGULATIONS Revised Total Coliform Rule § 141.860 Violations. (a) E. coli MCL Violation. A system is in violation of the MCL for E. coli when any of the conditions identified in paragraphs (a)(1) through (a)(4) of this section occur. (1) The system has an E. coli-positive repeat sample...

Self-Harm and Conventional Gender Roles in Women

ERIC Educational Resources Information Center

Straiton, Melanie L.; Hjelmeland, Heidi; Grimholt, Tine K.; Dieserud, Gudrun

2013-01-01

A total of thirty-two women admitted to a general hospital for medical treatment after self-harming completed measures of conventional positive and negative masculinity and femininity. Comparisons were made with two control groups with no self-harm history; 33 women receiving psychiatric outpatient treatment and a nonclinical sample of 206 women.…

Self-Informant Agreement in Well-Being Ratings: A Meta-Analysis

ERIC Educational Resources Information Center

Schneider, Leann; Schimmack, Ulrich

2009-01-01

A meta-analysis of published studies that reported correlations between self-ratings and informant ratings of well-being (life-satisfaction, happiness, positive affect, negative affect) was performed. The average self-informant correlation based on 44 independent samples and 81 correlations for a total of 8,897 participants was r = 0.42 [99%…

Molecular identification of Enterocytozoon bieneusi, Cryptosporidium, and Giardia in brazilian captive birds

USDA-ARS?s Scientific Manuscript database

A total of 85 fecal samples from captive birds collected from October 2013 to September 2014 in Uberlândia and Belo Horizonte in the state of Minas Gerais (Brazil) were evaluated for the presence of Enterocytozoon bieneusi, Cryptosporidium, and Giardia by PCR. Of these, 3 birds were found positive f...

Development of an on-site screening system for amphetamine-type stimulant tablets with a portable attenuated total reflection Fourier transform infrared spectrometer.

PubMed

Tsujikawa, Kenji; Kuwayama, Kenji; Miyaguchi, Hajime; Kanamori, Tatsuyuki; Iwata, Yuko T; Yoshida, Takemi; Inoue, Hiroyuki

2008-02-04

We tried to develop a library search system using a portable, attenuated total reflection Fourier transform infrared (ATR-FT-IR) spectrometer for on-site identification of 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA) tablets. The library consisted of the spectra from mixtures of controlled drugs (e.g. MDMA and ketamine), adulterants (e.g. caffeine), and diluents (e.g. lactose). In the seven library search algorithms, the derivative correlation coefficient showed the best discriminant capability. This was enhanced by segmentation of the search area. The optimized search algorithm was validated by the positive (n=154, e.g. the standard mixtures containing the controlled drug, and the MDMA/MDA tablets confiscated) and negative samples (n=56, e.g. medicinal tablets). All validation samples except for four were judged truly. Final criteria for positive identification were decided on the basis of the results of the validation. In conclusion, a portable ATR-FT-IR spectrometer with our library search system would be a useful tool for on-site identification of amphetamine-type stimulant tablets.

SAPOVIRUSES IN CHILDREN WITH ACUTE GASTROENTERITIS FROM MANAUS , AMAZON REGION, BRAZIL, 2010-2011

PubMed Central

REYMÃO, Tammy Kathlyn Amaral; HERNANDEZ, Juliana das Merces; da COSTA, Samya Thalita Picanço; de SOUSA, Maísa Silva; OLIVEIRA, Darleise de Souza; da SILVA, Luciana Damascena; BANDEIRA, Renato da Silva; de LIMA, Ian Carlos Gomes; SOARES, Luana da Silva; MASCARENHAS, Joana Darc Pereira; GABBAY, Yvone Benchimol

2016-01-01

SUMMARY Sapoviruses (SaVs) are responsible for acute gastroenteritis in humans, especially children and the elderly. In Brazil, data on SaVs infections are very limited, especially in Northern Brazil. Here, we investigated the occurrence of SaVs in samples from hospitalized children under ten years old that presented acute gastroenteritis. Positive samples were genotyped and phylogenetic analysis was performed using prototype strains sequences obtained from GenBank database. In total, 156 fecal samples were screened by RT-PCR for SaVs. A positivity rate of 3.8% (6/156) was found in children under three years of age. Four genotypes were detected: GI.I, GI.2 and GII.2?-GII.4?/GII.4, suggesting a possible inter-genotypes recombination. Most infections (83.3%) occurred between August and September. The positivity was similar to that found in other countries and genotyping demonstrated the presence of distinct genotypes. To our knowledge, this is the first study reporting the circulation of SaVs in Manaus, state of Amazonas, Amazon region, Brazil. PMID:27828622

The epidemiology of infections with Giardia species and genotypes in well cared for dogs and cats in Germany.

PubMed

Pallant, Louise; Barutzki, Dieter; Schaper, Roland; Thompson, R C Andrew

2015-01-06

Giardia is now considered the most common enteric parasite in well cared for dogs and cats in developed countries. The ecology, epidemiology and clinical impact of infections with this parasite in such animals is still not fully understood due to variable results across different studies. Faecal samples were collected between 2009 and 2012 from privately owned cats and dogs in Germany presented to local veterinarians for a variety of reasons. Giardia positive samples were identified by microscopy and coproantigen methods. Total faecal DNA was extracted from Giardia positive samples and multilocus genotyping methods (18S rDNA, β-giardin, GDH) were applied. Relationships between host age, sex, and breed, season of presentation and the different species of Giardia detected were assessed. A total of 60 cat and 130 dog samples were identified as Giardia positive. Potentially zoonotic Giardia was identified in both animal species. Cats had a similarly high rate of infection with the G. duodenalis and G. cati. Cats less than 1 year were more likely to have G. duodenalis than cats older than 1 year. Pure breed cats demonstrated a greater proportion of zoonotic species than mixed breed cats. In samples from dogs, G. canis (C and D genotypes) were identified most commonly. Male dogs were more likely to have G. canis (genotype D) than female dogs. The 18S rDNA PCR protocol was the most successful followed by the β-giardin and GDH (amplifying from 92%, 42% and 13% of samples respectively). The potentially zoonotic species G. duodenalis and G. enterica were found in cat and dog samples, with G. duodenalis found in greater numbers; however, this may be due to the detection techniques utilised. Cats appeared to show a relationship between G. duodenalis and G. cati with age and breed, which may be explained by different housing habitats for pure and mixed breed cats. The different success rates for the three loci utilised highlights the usefulness of the 18S locus as a screening tool, as well as the importance of using multiple loci for genotyping to fully determine the level of multiple infection of Giardia present.

Identical Assemblage of Giardia duodenalis in Humans, Animals and Vegetables in an Urban Area in Southern Brazil Indicates a Relationship among Them

PubMed Central

Colli, Cristiane Maria; Bezagio, Renata Coltro; Nishi, Letícia; Bignotto, Thaís Souto; Ferreira, Érika Cristina; Falavigna-Guilherme, Ana Lúcia; Gomes, Mônica Lúcia

2015-01-01

Background Giardia duodenalis infects humans and other mammals by ingestion of cysts in contaminated water or food, or directly in environments with poor hygiene. Eight assemblages, designated A–H, are described for this species. Methodology/Principal Findings We investigated by microscopy or by direct immunofluorescence technique the occurrence of G. duodenalis in 380 humans, 34 animals, 44 samples of water and 11 of vegetables. G. duodenalis cysts present in samples were genotyped through PCR-RFLP of β giardin and glutamate dehydrogenase (gdh) genes and sequencing of gdh. The gdh gene was amplified in 76.5% (26/34) of the human faeces samples with positive microscopy and in 2.9% (1/34) of negative samples. In 70.4% (19/27) of the positive samples were found BIV assemblage. In two samples from dogs with positive microscopy and one negative sample, assemblages BIV, C, and D were found. Cysts of Giardia were not detected in water samples, but three samples used for vegetable irrigation showed total coliforms above the allowed limit, and Escherichia coli was observed in one sample. G. duodenalis BIV was detected in two samples of Lactuca sativa irrigated with this sample of water. BIV was a common genotype, with 100% similarity, between different sources or hosts (humans, animals and vegetables), and the one most often found in humans. Conclusions/Significance This is the first study in Brazil that reports the connection among humans, dogs and vegetables in the transmission dynamics of G. duodenalis in the same geographic area finding identical assemblage. BIV assemblage was the most frequently observed among these different links in the epidemiological chain. PMID:25761119

Detection and clearance of prostate cells subsequent to ultrasound-guided needle biopsy as determined by multiplex nested reverse transcription polymerase chain reaction assay.

PubMed

Price, D K; Clontz, D R; Woodard, W L; Kaufman, J S; Daniels, J M; Stolzenberg, S J; Teigland, C M

1998-08-01

To determine if circulating prostate cells are detectable subsequent to transrectal ultrasound (TRUS)-guided biopsy, and if so, whether cells remain in circulation for up to 4 weeks. Blood samples were drawn from 90 patients with elevated serum prostate-specific antigen (PSA) levels and/or abnormal digital rectal examination. Two samples were drawn from all patients immediately prior to TRUS and 30 minutes postbiopsy. Blood samples were also obtained 1 week postbiopsy from 83 patients, and 1 month postbiopsy from 61 patients. Multiplex nested reverse transcription polymerase chain reaction assay (RT-PCR) for PSA and prostate-specific membrane antigen (PSM) was performed on total ribonucleic acid (RNA) from each sample. Results were reported as positive if at least one of the targets was detected. Of 45 patients with biopsy-proven adenocarcinoma, 22 were RT-PCR positive prebiopsy and all remained positive 30 minutes postbiopsy. Of 23 patients with adenocarcinoma who were RT-PCR negative prebiopsy, 5 (22%) converted to positive 30 minutes postbiopsy (P < 0.001). Four of these 5 patients returned to negative after 1 week or 1 month. Of 45 patients without cancer at biopsy, 32 were RT-PCR negative prebiopsy and 6 (19%) converted to positive 30 minutes postbiopsy (P < 0.001). Although four of six available samples were still positive at 1 week, all four samples available 1 month postbiopsy were negative. Detection of circulating prostate cells subsequent to biopsy occurred in 11 of 55 (20%) previously RT-PCR negative patients, a proportion twice that reported in the literature. We attribute this higher proportion to the simultaneous detection of PSA and PSM mRNA in our multiplex assay. Conversion rates were similar in patients regardless of biopsy result. Testing of serial postbiopsy blood demonstrates clearing of these cells by 4 weeks in most patients.

Species richness of arbuscular mycorrhizal fungi: associations with grassland plant richness and biomass.

PubMed

Hiiesalu, Inga; Pärtel, Meelis; Davison, John; Gerhold, Pille; Metsis, Madis; Moora, Mari; Öpik, Maarja; Vasar, Martti; Zobel, Martin; Wilson, Scott D

2014-07-01

Although experiments show a positive association between vascular plant and arbuscular mycorrhizal fungal (AMF) species richness, evidence from natural ecosystems is scarce. Furthermore, there is little knowledge about how AMF richness varies with belowground plant richness and biomass. We examined relationships among AMF richness, above- and belowground plant richness, and plant root and shoot biomass in a native North American grassland. Root-colonizing AMF richness and belowground plant richness were detected from the same bulk root samples by 454-sequencing of the AMF SSU rRNA and plant trnL genes. In total we detected 63 AMF taxa. Plant richness was 1.5 times greater belowground than aboveground. AMF richness was significantly positively correlated with plant species richness, and more strongly with below- than aboveground plant richness. Belowground plant richness was positively correlated with belowground plant biomass and total plant biomass, whereas aboveground plant richness was positively correlated only with belowground plant biomass. By contrast, AMF richness was negatively correlated with belowground and total plant biomass. Our results indicate that AMF richness and plant belowground richness are more strongly related with each other and with plant community biomass than with the plant aboveground richness measures that have been almost exclusively considered to date. © 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.

Groundwater arsenic in Chimaltenango, Guatemala.

PubMed

Lotter, Jason T; Lacey, Steven E; Lopez, Ramon; Socoy Set, Genaro; Khodadoust, Amid P; Erdal, Serap

2014-09-01

In the Municipality of Chimaltenango, Guatemala, we sampled groundwater for total inorganic arsenic. In total, 42 samples were collected from 27 (43.5%) of the 62 wells in the municipality, with sites chosen to achieve spatial representation throughout the municipality. Samples were collected from household faucets used for drinking water, and sent to the USA for analysis. The only site found to have a concentration above the 10 μg/L World Health Organization provisional guideline for arsenic in drinking water was Cerro Alto, where the average concentration was 47.5 μg/L. A health risk assessment based on the arsenic levels found in Cerro Alto showed an increase in noncarcinogenic and carcinogenic risks for residents as a result of consuming groundwater as their primary drinking water source. Using data from the US Geological Survey and our global positioning system data of the sample locations, we found Cerro Alto to be the only site sampled within the tertiary volcanic rock layer, a known source of naturally occurring arsenic. Recommendations were made to reduce the levels of arsenic found in the community's drinking water so that the health risks can be managed.

Rapid identification of bacteria from positive blood culture bottles by use of matrix-assisted laser desorption-ionization time of flight mass spectrometry fingerprinting.

PubMed

Christner, Martin; Rohde, Holger; Wolters, Manuel; Sobottka, Ingo; Wegscheider, Karl; Aepfelbacher, Martin

2010-05-01

Early and adequate antimicrobial therapy has been shown to improve the clinical outcome in bloodstream infections (BSI). To provide rapid pathogen identification for targeted treatment, we applied matrix-assisted laser desorption-ionization time of flight (MALDI-TOF) mass spectrometry fingerprinting to bacteria directly recovered from blood culture bottles. A total of 304 aerobic and anaerobic blood cultures, reported positive by a Bactec 9240 system, were subjected in parallel to differential centrifugation with subsequent mass spectrometry fingerprinting and reference identification using established microbiological methods. A representative spectrum of bloodstream pathogens was recovered from 277 samples that grew a single bacterial isolate. Species identification by direct mass spectrometry fingerprinting matched reference identification in 95% of these samples and worked equally well for aerobic and anaerobic culture bottles. Application of commonly used score cutoffs to classify the fingerprinting results led to an identification rate of 87%. Mismatching mostly resulted from insufficient bacterial numbers and preferentially occurred with Gram-positive samples. The respective spectra showed low concordance to database references and were effectively rejected by score thresholds. Spiking experiments and examination of the respective study samples even suggested applicability of the method to mixed cultures. With turnaround times around 100 min, the approach allowed for reliable pathogen identification at the day of blood culture positivity, providing treatment-relevant information within the critical phase of septic illness.

Clinical comparison of the Treponema pallidum CAPTIA syphilis-G enzyme immunoassay with the fluorescent treponemal antibody absorption immunoglobulin G assay for syphilis testing.

PubMed

Halling, V W; Jones, M F; Bestrom, J E; Wold, A D; Rosenblatt, J E; Smith, T F; Cockerill, F R

1999-10-01

Recently, a treponema-specific immunoglobulin G (IgG) enzyme immunoassay (EIA), the CAPTIA Syphilis-G (Trinity Biotech, Jamestown, N.Y.), has become available as a diagnostic test for syphilis. A total of 89 stored sera previously tested by the fluorescent treponemal antibody absorption (FTA-ABS) IgG assay were evaluated by the CAPTIA EIA. The FTA-ABS IgG procedure was performed by technologists unblinded to results of rapid plasmid reagin (RPR) testing of the same specimens. Borderline CAPTIA-positive samples (antibody indices of >/=0.650 and 0.900, the sample was considered positive. Thirteen of 89 (15%) samples had discrepant results. Compared to the FTA-ABS assay, the CAPTIA EIA had a sensitivity and specificity and positive and negative predictive values of 70.7, 97.9, 96.7, and 79.7%, respectively. In another analysis, discrepancies between results were resolved by repeated FTA-ABS testing (technologists were blinded to previous RPR results) and patient chart reviews. Seven CAPTIA-negative samples which were previously interpreted (unblinded) as minimally reactive by the FTA method were subsequently interpreted (blinded) as nonreactive. One other discrepant sample (CAPTIA negative and FTA-ABS positive [at an intensity of 3+], unblinded) was FTA negative with repeated testing (blinded). For the five remaining discrepant samples, chart reviews indicated that one patient (CAPTIA negative and FTA-ABS positive [minimally reactive], blinded) had possible syphilis. These five samples were also evaluated and found to be negative by another treponema-specific test, the Treponema pallidum microhemagglutination assay. Therefore, after repeated testing and chart reviews, 2 of the 89 (2%) samples had discrepant results; the adjusted sensitivity, specificity, and positive and negative predictive values were 96.7, 98.3, 96.7, and 98.3%, respectively. This study demonstrates that the CAPTIA IgG EIA is a reliable method for syphilis testing and that personnel performing tests which require subjective interpretation, like the FTA-ABS test, may be biased by RPR test results.

Abattoir based surveillance of cystic echinococcosis (CE) in the Sultanate of Oman during 2010-2013.

PubMed

Al Kitani, Fadya Abdullah; Al Riyami, Shumoos; Al Yahyai, Sabra; Al Rawahi, Abdulmajeed Hamood; Al Maawali, Mahir; Al Awahi, Abdulmajeed Hamood; Al Aawali, Mahir; Hussain, Muhammad Hammad

2015-07-30

A passive surveillance was conducted in 8 selected abattoirs belonging to 8 governorates of Oman. Routine slaughtering of 282.020 animals was observed during 2010-2013 by duty veterinarians and suspected samples (n=1440) for cystic echinococcosis (CE) from camels (n=292), cattle (n=303), goats (n=682) and sheep (n=163) were collected. In total, 587 (40.8%) samples were found positive for CE and the highest percentage (64.2%) of positive samples belonged to Dhofar (Salalah) as compared to other governorates, p<0.01. The infection rate of CE was calculated as 5.3, 0.6, 0.07 and 0.03 in camel, cattle, sheep and goats respectively. Fertile cysts were found in 30.7% of positive samples and camels (52%) were found as major source of these followed by cattle (14.1%) and goats (9.7%). Percentage of positive to submitted samples was higher in females (67.9%) than males (29.8%). The fertility of cyst increased with age and the highest percentage of fertile cysts were collected from animals above 5 years of age (54.4%) followed by between 3 and 5 years (36.7%) and below 3 years of age (8.9%). For different breeds, 478 (62.7%) samples from local breeds of livestock were positive for CE as compared to 108 (15.9%) from imported animals (OR 8.92), p<0.01. The highest percent of positive samples was observed in cysts from lungs (54.4%) followed by those from liver (25.7%), liver and lungs (14.1%) and others (3.8%). Molecular investigations indicated that G1 and G6/7 were the two strains of CE circulating in camels and cattle while G1 was the only strain found in samples from small ruminants. Results indicated that CE is endemic in the livestock of Oman and further molecular epidemiological work in different host species is required for the conceptualization of a comprehensive control program. Copyright © 2015 Elsevier B.V. All rights reserved.

Clinical Comparison of the Treponema pallidum CAPTIA Syphilis-G Enzyme Immunoassay with the Fluorescent Treponemal Antibody Absorption Immunoglobulin G Assay for Syphilis Testing

PubMed Central

Halling, V. W.; Jones, M. F.; Bestrom, J. E.; Wold, A. D.; Rosenblatt, J. E.; Smith, T. F.; Cockerill, F. R.

1999-01-01

Recently, a treponema-specific immunoglobulin G (IgG) enzyme immunoassay (EIA), the CAPTIA Syphilis-G (Trinity Biotech, Jamestown, N.Y.), has become available as a diagnostic test for syphilis. A total of 89 stored sera previously tested by the fluorescent treponemal antibody absorption (FTA-ABS) IgG assay were evaluated by the CAPTIA EIA. The FTA-ABS IgG procedure was performed by technologists unblinded to results of rapid plasmid reagin (RPR) testing of the same specimens. Borderline CAPTIA-positive samples (antibody indices of ≥0.650 and ≤0.900) were retested; if the second analysis produced an index of >0.900, the sample was considered positive. Thirteen of 89 (15%) samples had discrepant results. Compared to the FTA-ABS assay, the CAPTIA EIA had a sensitivity and specificity and positive and negative predictive values of 70.7, 97.9, 96.7, and 79.7%, respectively. In another analysis, discrepancies between results were resolved by repeated FTA-ABS testing (technologists were blinded to previous RPR results) and patient chart reviews. Seven CAPTIA-negative samples which were previously interpreted (unblinded) as minimally reactive by the FTA method were subsequently interpreted (blinded) as nonreactive. One other discrepant sample (CAPTIA negative and FTA-ABS positive [at an intensity of 3+], unblinded) was FTA negative with repeated testing (blinded). For the five remaining discrepant samples, chart reviews indicated that one patient (CAPTIA negative and FTA-ABS positive [minimally reactive], blinded) had possible syphilis. These five samples were also evaluated and found to be negative by another treponema-specific test, the Treponema pallidum microhemagglutination assay. Therefore, after repeated testing and chart reviews, 2 of the 89 (2%) samples had discrepant results; the adjusted sensitivity, specificity, and positive and negative predictive values were 96.7, 98.3, 96.7, and 98.3%, respectively. This study demonstrates that the CAPTIA IgG EIA is a reliable method for syphilis testing and that personnel performing tests which require subjective interpretation, like the FTA-ABS test, may be biased by RPR test results. PMID:10488183

Vulnerability of drinking-water wells in La Crosse, Wisconsin, to enteric-virus contamination from surface water contributions.

PubMed

Borchardt, Mark A; Haas, Nathaniel L; Hunt, Randall J

2004-10-01

Human enteric viruses can contaminate municipal drinking-water wells, but few studies have examined the routes by which viruses enter these wells. In the present study, the objective was to monitor the municipal wells of La Crosse, Wisconsin, for enteric viruses and determine whether the amount of Mississippi River water infiltrating the wells was related to the frequency of virus detection. From March 2001 to February 2002, one river water site and four wells predicted by hydrogeological modeling to have variable degrees of surface water contributions were sampled monthly for enteric viruses, microbial indicators of sanitary quality, and oxygen and hydrogen isotopes. (18)O/(16)O and (2)H/(1)H ratios were used to determine the level of surface water contributions. All samples were collected prior to chlorination at the wellhead. By reverse transcription-PCR (RT-PCR), 24 of 48 municipal well water samples (50%) were positive for enteric viruses, including enteroviruses, rotavirus, hepatitis A virus (HAV), and noroviruses. Of 12 river water samples, 10 (83%) were virus positive by RT-PCR. Viable enteroviruses were not detected by cell culture in the well samples, although three well samples were positive for culturable HAV. Enteroviruses detected in the wells by RT-PCR were identified as several serotypes of echoviruses and group A and group B coxsackieviruses. None of the well water samples was positive for indicators of sanitary quality, namely male-specific and somatic coliphages, total coliform bacteria, Escherichia coli, and fecal enterococci. Contrary to expectations, viruses were found in all wells regardless of the level of surface water contributions. This result suggests that there were other unidentified sources, in addition to surface water, responsible for the contamination.

Indeterminate RIBA results were associated with the absence of hepatitis C virus RNA (HCV-RNA) in blood donors.

PubMed

Pereira, Felicidade Mota; Zarife, Maria Alice Sant'ana; Reis, Eliana Almeida Gomes; G Reis, Mitermayer

2014-01-01

Hepatitis C virus (HCV) infection is diagnosed by the presence of antibodies and is supplemented by confirmatory testing methods, such as recombinant immunoblot assay (RIBA) and HCV-RNA detection. This study aimed to evaluate the efficacy of RIBA testing to diagnose HCV infection in blood donors positive for anti-HCV antibodies. A total of 102 subjects positive for anti-HCV determined by enzyme-linked immunosorbent assay (ELISA) at the Hematology and Hemotherapy Foundation of Bahia (HEMOBA) were later assessed with new samples using the Abbott Architect anti-HCV test (Abbott Diagnostics, Wiesbaden, Germany), the RIBA III test (Chiron RIBA HCV 3.0 SIA, Chiron Corp., Emeryville, CA, USA), the polymerase chain reaction (PCR; COBAS® AMPLICOR HCV Roche Diagnostics Corp., Indianapolis, IN, USA) and line probe assay (LiPA - Siemens, Tarrytown, NY, USA) genotyping for HCV diagnosis. Of these new samples, 38.2% (39/102) were positive, 57.8% (59/102) were negative and 3.9% (4/102) were indeterminate for anti-HCV; HCV-RNA was detected in 22.5% (23/102) of the samples. RIBA results were positive in 58.1% (25/43), negative in 9.3% (4/43) and indeterminate in 32.6% (14/43) of the samples. The prevailing genotypes were 1 (78.3%, 18/23), 3 (17.4%, 4/23) and 2 (4.3%, 1/23). All 14 samples with indeterminate RIBA results had undetectable viral loads (detection limit ≤50 IU/mL). Of these samples, 71.4% (10/14) were reevaluated six months later. Eighty percent (8/10) of these samples remained indeterminate by RIBA, and 20% (2/10) were negative. In this study, individuals with indeterminate RIBA results had no detectable HCV-RNA.

Vulnerability of Drinking-Water Wells in La Crosse, Wisconsin, to Enteric-Virus Contamination from Surface Water Contributions

PubMed Central

Borchardt, Mark A.; Haas, Nathaniel L.; Hunt, Randall J.

2004-01-01

Human enteric viruses can contaminate municipal drinking-water wells, but few studies have examined the routes by which viruses enter these wells. In the present study, the objective was to monitor the municipal wells of La Crosse, Wisconsin, for enteric viruses and determine whether the amount of Mississippi River water infiltrating the wells was related to the frequency of virus detection. From March 2001 to February 2002, one river water site and four wells predicted by hydrogeological modeling to have variable degrees of surface water contributions were sampled monthly for enteric viruses, microbial indicators of sanitary quality, and oxygen and hydrogen isotopes. 18O/16O and 2H/1H ratios were used to determine the level of surface water contributions. All samples were collected prior to chlorination at the wellhead. By reverse transcription-PCR (RT-PCR), 24 of 48 municipal well water samples (50%) were positive for enteric viruses, including enteroviruses, rotavirus, hepatitis A virus (HAV), and noroviruses. Of 12 river water samples, 10 (83%) were virus positive by RT-PCR. Viable enteroviruses were not detected by cell culture in the well samples, although three well samples were positive for culturable HAV. Enteroviruses detected in the wells by RT-PCR were identified as several serotypes of echoviruses and group A and group B coxsackieviruses. None of the well water samples was positive for indicators of sanitary quality, namely male-specific and somatic coliphages, total coliform bacteria, Escherichia coli, and fecal enterococci. Contrary to expectations, viruses were found in all wells regardless of the level of surface water contributions. This result suggests that there were other unidentified sources, in addition to surface water, responsible for the contamination. PMID:15466536

Vulnerability of drinking-water wells in La Crosse, Wisconsin, to enteric-virus contamination from surface water contributions

USGS Publications Warehouse

Borchardt, M. A.; Haas, N.L.; Hunt, R.J.

2004-01-01

Human enteric viruses can contaminate municipal drinking-water wells, but few studies have examined the routes by which viruses enter these wells. In the present study, the objective was to monitor the municipal wells of La Crosse, Wisconsin, for enteric viruses and determine whether the amount of Mississippi River water infiltrating the wells was related to the frequency of virus detection. From March 2001 to February 2002, one river water site and four wells predicted by hydrogeological modeling to have variable degrees of surface water contributions were sampled monthly for enteric viruses, microbial indicators of sanitary quality, and oxygen and hydrogen isotopes. 18O/ 16O and 2H/1H ratios were used to determine the level of surface water contributions. All samples were collected prior to chlorination at the wellhead. By reverse transcription-PCR (RT-PCR), 24 of 48 municipal well water samples (50%) were positive for enteric viruses, including enteroviruses, rotavirus, hepatitis A virus (HAV), and noroviruses. Of 12 river water samples, 10 (83%) were virus positive by RT-PCR. Viable enteroviruses were not detected by cell culture in the well samples, although three well samples were positive for culturable HAV. Enteroviruses detected in the wells by RT-PCR were identified as several serotypes of echoviruses and group A and group B coxsackieviruses. None of the well water samples was positive for indicators of sanitary quality, namely male-specific and somatic coliphages, total coliform bacteria, Escherichia coli, and fecal enterococci. Contrary to expectations, viruses were found in all wells regardless of the level of surface water contributions. This result suggests that there were other unidentified sources, in addition to surface water, responsible for the contamination.

Is repetitive intraoperative splash basin use a source of bacterial contamination in total joint replacement?

PubMed

Glait, Sergio A; Schwarzkopf, Ran; Gould, Steven; Bosco, Joseph; Slover, James

2011-09-09

Splash basins are used in arthroplasty cases to wash instruments. Several studies in the literature have shown these basins being a potential source of bacterial infection. This study assesses the risk of contamination of intraoperative splash basins used to wash and store instruments. A total of 46 random clean primary arthroplasty cases (32 hips, 13 knees, and 1 unicondylar knee) were studied by taking cultures of sterile splash basins as soon as they are opened (controls) and again at wound closure after instruments and debris have come into contact with the sterile water. All cultures were taken with sterile culture swabs and sent to the laboratory for aerobic, anaerobic, and fungal culture. Outcome measured was any positive culture. A total of 92 cultures from 46 cases were tested. Only 1 (2.17%) control culture, which grew Streptococcus viridans, was positive for bacterial growth. One of 46 samples (2.17%) taken at wound closure was positive for coagulase-negative Staphylococcus. Mean time between basin opening and wound closure was 180±45 minutes. For the 1 infected sample taken at the conclusion of the case, it was 240 minutes. Previous studies show contamination rates as high as 74% for splash basins used intraoperatively. Our study contradicts the belief that splash basins are a high source of infection, with only 2.17% of basins showing contamination. Splash basins can be a potential source of contamination, but the risk is not as high as previously cited in the orthopedic literature. Copyright 2011, SLACK Incorporated.

Isolation of microbial pathogens of subclinical mastitis from raw sheep's milk of Epirus (Greece) and their role in its hygiene.

PubMed

Fotou, K; Tzora, A; Voidarou, Ch; Alexopoulos, A; Plessas, S; Avgeris, I; Bezirtzoglou, E; Akrida-Demertzi, K; Demertzis, P G

2011-12-01

The natural raw milk microflora is a factor that expresses its sensorial characteristics. The microbial charge into the mammary gland of healthy animal is low and the application of right and healthy conditions during milking and cheese making procedure, prevents from contaminating as well as maintains the natural microflora in order to lend the particular characteristics of milk. The purpose of the present project was the study of the Total Viable Count (T.V.C.) and the count of total psychrotropic bacteria of raw sheep milk from Boutsiko and Karamaniko breeds, collected from healthy animals, as well as the isolation, identification and enumeration of pathogenic bacteria related with the hygiene and the quality of raw sheep milk (with a particular interest in bacteria that may cause human infection). During the experiment we examined two hundred forty (240) samples of raw sheep milk. In these samples a) Staphylococcus aureus, Salmonella sp., Escherichia coli, Clostridium perfringens (vegetative cells and spores) and Bacillus sp. were isolated and identified b) the Total Viable Count and the total number of psychrotropic bacteria were also specified. The sampling, the preparation of samples and decimal dilutions were based on international methods. The Total viable count was determined using the standard methods of the American Public Health Association, 2002. The total number of psychrotropic bacteria was determined using APHA 1976, 1978 rules. The identification of the bacteria was carried out according to the Bergey's manual. Microscopic examination of Gram stained cells, catalase, oxidase and biochemical tests were performed when necessary to further identify. From the 240 milk samples tested, only 5% were E. coli positive, with mean counts ranged from 2 × 10(3) to 2.4 × 10(4) cfu/ml. S. aureus was isolated from 24% of the samples and the mean count per ml was ranged from <10 to 3.4 × 10(2). Meanwhile, Bacillus spp. was also detected in 29% samples. Vegetative forms and spores of C. perfringens were detected in 13% and 63% of the samples respectively. However, microbiological analyses revealed the presence of a small number of selected pathogens in milk samples such as Salmonella, which was only detected in 5% of the samples. Listeria sp., Pseudomonas sp. and Vibrio cholerae were never found. From the experimental results, the Total Viable Count from raw sheep milk samples, fulfils the microbiological criteria of EU Legislation in a percentage of approximately 97%. Copyright © 2011 Elsevier Ltd. All rights reserved.

Environmental monitoring and analysis of faecal contamination in an urban setting in the city of Bari (Apulia region, Italy): health and hygiene implications.

PubMed

Tarsitano, Elvira; Greco, Grazia; Decaro, Nicola; Nicassio, Francesco; Lucente, Maria Stella; Buonavoglia, Canio; Tempesta, Maria

2010-11-01

Few studies have been conducted in Italy to quantify the potential risk associated with dynamics and distribution of pathogens in urban settings. The aim of this study was to acquire data on the environmental faecal contamination in urban ecosystems, by assessing the presence of pathogens in public areas in the city of Bari (Apulia region, Italy). To determine the degree of environmental contamination, samples of dog faeces and bird guano were collected from different areas in the city of Bari (park green areas, playgrounds, public housing areas, parkways, and a school). A total of 152 canine faecal samples, in 54 pools, and two samples of pigeon guano from 66 monitored sites were examined. No samples were found in 12 areas spread over nine sites. Chlamydophila psittaci was detected in seven canine and two pigeon guano samples. Salmonella species were not found. On the other hand, four of 54 canine faecal samples were positive for reovirus. Thirteen canine faecal samples were positive for parasite eggs: 8/54 samples contained Toxocara canis and Toxascaris leonina eggs and 5/54 samples contained Ancylostoma caninum eggs. Our study showed that public areas are often contaminated by potentially zoonotic pathogens.

Effect of season and ambient temperature on outcome of guaiac-based faecal occult blood tests performed for colorectal cancer screening.

PubMed

Hunter, J P; Saratzis, A; Froggatt, P; Harmston, C

2012-09-01

Guaiac-based faecal occult blood tests (gFOBTs) are used in the colorectal cancer screening programme. Recent data suggested that the immunological faecal occult blood test illustrated a variation in positivity according to season and ambient temperature. Our aim was to assess the effect of season and ambient temperature on the positivity rates of the gFOBT during pilot screening for colorectal cancer. Data from the first year of round 1 of the pilot screening programme in Coventry and Warwickshire were analysed. Patients with positive and negative gFOBT samples were included. Patients with spoilt samples or incomplete data were excluded. Of the total of 59513 patients, 30311 were men and 29202 women. Mean age was 56 years. Daily temperature data were provided by the meteorological office. Median exposure of the gFOBT test card was 6 days (range 1-17). Median daily maximum temperature was 14°C. Spring and summer illustrated significantly decreased positivity rates compared with autumn and winter (Pearson's chi-squared test, P<0.001). Mean daily maximum temperature for the test card exposure showed no significant difference in positivity rates (P=0.53). Subgroup analysis revealed a significant reduction in positive samples in the >25°C subgroup (P=0.045). There is a seasonal variation in positivity rates of gFOBTs with increased positivity in spring and summer months. There is no difference in positivity rates in relation to ambient temperature except in subgroup analysis where there is a significant reduction in positivity rates above 25°C. © 2011 The Authors. Colorectal Disease © 2011 The Association of Coloproctology of Great Britain and Ireland.

[Genetic profiling of Giardia intestinalis by polimerase chain in human and dogs samples of Colombian Caribean Coast].

PubMed

Arroyo-Salgado, Bárbara; Buelvas-Montes, Yaleyvis; Villalba-Vizcaíno, Vivian; Salomón-Arzuza, Octavio

2014-01-01

Giardia intestinalis (G. Intestinalis) is a protozoan that causes diarrheal disease and malabsorption syndrome in humans and other mammals. It presents a high genetic diversity evidenced in the recognition of 7 genotypes (A-G). Genotypes A and B are commonly associated to humans and domestic animals such as dogs. The aim of this study was to conduct a preliminary genetic characterization of G. intestinalis in humans and dogs from two cities on the Caribbean coast of Colombia. Sampling areas were selected according to the highest numbers of acute diarrheal disease. Stool samples were collected from children under 7 years old, with positive medical tests for G. intestinalis. Cysts were purified by sucrose gradient and DNA samples were isolated by extraction with organic solvents. Molecular characterization was performed by amplifying the gene triose phosphate isomerase (tpi) by using a semi-nested PCR. A total of 202 samples of DNA were obtained; of these, 111 were positive in coproparasitological analysis (13 dogs and 98 children). Genotype distribution in positive samples was: 5.1% belonged to genotype A and 92.3% to genotype B. Genotype B was present in humans and animals. The most common genotype in both human and animal samples was genotype B, suggesting a zoonotic transmission cycle. Copyright © 2012 Elsevier España, S.L.U. y Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

Spatial distribution of enteric viruses and somatic coliphages in a Lagoon used as drinking water source and recreation in Southern Brazil.

PubMed

Elmahdy, M E I; Fongaro, G; Magri, M E; Petruccio, M M; Barardi, C R M

2016-10-01

This study aimed to evaluate the contamination level of the Peri Lagoon, the main freshwater reservoir of Santa Catarina Island, Southern Brazil, for human adenovirus (HAdV), hepatitis A virus (HAV), rotavirus species A (RVA), and somatic coliphages (SOMCPH). Viruses were also investigated in sediments and their sensitivity against natural sunlight was analysed by studying their spatial distribution in different depths of the water column. A total of 84 water samples and 48 sediment samples were examined by qPCR or RT-qPCR. Infectivity of HAdV and SOMCPH was determined and quantified by plaque assay method. A sum of 64% and 48% of water and sediment samples were positive for HAdV, respectively. RVA was present in 33% and 18% of water and sediment samples, and 25% of water samples were positive for HAV. HAdV were infectious in 76% of water and 83% of sediment samples that were positive by qPCR. SOMCPH could be detected in 42% and 18% of water and sediment samples, respectively. The data pointed a variation of viruses' prevalence according to the different water column depths. These results demonstrated that water sources and sediments contaminated by human wastes could play an important role in the recontamination of water columns harvested for further treatment or used for recreational purposes. These data can be of great value for future risk assessment analysis. Copyright © 2016. Published by Elsevier GmbH.

Microbial and Antibiotic Susceptibility Profile among Clinical Samples of Patients with Acute Leukemia.

PubMed

Abdollahi, Alireza; Hakimi, Faezeh; Doomanlou, Mahsa; Azadegan, Azadeh

2016-04-01

Preventing and starting early treatment of infections in patients whose immunity system is weak due to malignancies like leukemia can reduce mortality. This study aimed to determine microbial and antibiotic resistance patterns in clinical samples of patients with acute leukemia to start early treatment before the results of clinical tests are known. In this cross-sectional study, the clinical samples of all patients hospitalized with the diagnosis of acute leukemia were cultured and their antibiogram was evaluated. Then, the data were analyzed by SPSS 18 based on the objectives of the study. Of a total of 2,366 samples, 18.95% were reported to be positive blood samples, 22.96% were reported to be urine samples and 36% wound samples. E. coli was the most common bacteria isolated from the blood and urine cultures (34% in blood, 32% in urine culture) while Staphylococcus Aureus was the most common in the wound culture (35%). The highest level of sensitivity in the organisms with positive blood culture was to Ciprofloxacin, while in positive urine and wound culture was to Imipenem. The highest resistance in blood, urine and wound culture was to Cotrimoxazole. According to results obtained from this study, it is necessary to conduct appropriate studies on this issue in specific conditions in our country. The findings of this study can be used in clinics for more accurate diagnosis, more effective treatment before the results of clinical tests are known and also for prevention of infection in cancer patients.

Relationship between presence of cows with milk positive for Mycobacterium avium subsp. paratuberculosis-specific antibody by enzyme-linked immunosorbent assay and viable M. avium subsp. paratuberculosis in dust in cattle barns.

PubMed

Eisenberg, Susanne W F; Chuchaisangrat, Ruj; Nielen, Mirjam; Koets, Ad P

2013-09-01

Paratuberculosis, or Johne's disease, in cattle is caused by Mycobacterium avium subsp. paratuberculosis, which has recently been suspected to be transmitted through dust. This longitudinal study on eight commercial M. avium subsp. paratuberculosis-positive dairy farms studied the relationship between the number of cows with M. avium subsp. paratuberculosis antibody-positive milk and the presence of viable M. avium subsp. paratuberculosis in settled-dust samples, including their temporal relationship. Milk and dust samples were collected in parallel monthly for 2 years. M. avium subsp. paratuberculosis antibodies in milk were measured by enzyme-linked immunosorbent assay (ELISA) and used as a proxy for M. avium subsp. paratuberculosis shedding. Settled-dust samples were collected by using electrostatic dust collectors (EDCs) at six locations in housing for dairy cattle and young stock. The presence of viable M. avium subsp. paratuberculosis was identified by liquid culture and PCR. The results showed a positive relationship (odds ratio [OR], 1.2) between the number of cows with ELISA-positive milk and the odds of having positive EDCs in the same airspace as the adult dairy cattle. Moreover, the total number of lactating cows also showed an OR slightly above 1. This relationship remained the same for settled-dust samples collected up to 2 months before or after the time of milk sampling. The results suggest that removal of adult cows with milk positive for M. avium subsp. paratuberculosis-specific antibody by ELISA might result in a decrease in the presence of viable M. avium subsp. paratuberculosis in dust and therefore in the environment. However, this decrease is likely delayed by several weeks at least. In addition, the data support the notion that M. avium subsp. paratuberculosis exposure of young stock is reduced by separate housing.

Relationship between Presence of Cows with Milk Positive for Mycobacterium avium subsp. paratuberculosis-Specific Antibody by Enzyme-Linked Immunosorbent Assay and Viable M. avium subsp. paratuberculosis in Dust in Cattle Barns

PubMed Central

Chuchaisangrat, Ruj; Nielen, Mirjam; Koets, Ad P.

2013-01-01

Paratuberculosis, or Johne's disease, in cattle is caused by Mycobacterium avium subsp. paratuberculosis, which has recently been suspected to be transmitted through dust. This longitudinal study on eight commercial M. avium subsp. paratuberculosis-positive dairy farms studied the relationship between the number of cows with M. avium subsp. paratuberculosis antibody-positive milk and the presence of viable M. avium subsp. paratuberculosis in settled-dust samples, including their temporal relationship. Milk and dust samples were collected in parallel monthly for 2 years. M. avium subsp. paratuberculosis antibodies in milk were measured by enzyme-linked immunosorbent assay (ELISA) and used as a proxy for M. avium subsp. paratuberculosis shedding. Settled-dust samples were collected by using electrostatic dust collectors (EDCs) at six locations in housing for dairy cattle and young stock. The presence of viable M. avium subsp. paratuberculosis was identified by liquid culture and PCR. The results showed a positive relationship (odds ratio [OR], 1.2) between the number of cows with ELISA-positive milk and the odds of having positive EDCs in the same airspace as the adult dairy cattle. Moreover, the total number of lactating cows also showed an OR slightly above 1. This relationship remained the same for settled-dust samples collected up to 2 months before or after the time of milk sampling. The results suggest that removal of adult cows with milk positive for M. avium subsp. paratuberculosis-specific antibody by ELISA might result in a decrease in the presence of viable M. avium subsp. paratuberculosis in dust and therefore in the environment. However, this decrease is likely delayed by several weeks at least. In addition, the data support the notion that M. avium subsp. paratuberculosis exposure of young stock is reduced by separate housing. PMID:23793639

Mycobacterium avium subsp. paratuberculosis from free-ranging deer and rabbits surrounding Minnesota dairy herds

PubMed Central

2005-01-01

Abstract The objectives of this study were to estimate the prevalence of Mycobacterium avium subsp. paratuberculosis (MAP) among deer and rabbits surrounding infected and noninfected Minnesota dairy farms using fecal culture, and to describe the frequency that farm management practices were used that could potentially lead to transmission of infection between these species. Fecal samples from cows and the cow environment were collected from 108 Minnesota dairy herds, and fecal pellets from free-ranging white-tailed deer and eastern cottontail rabbits were collected from locations surrounding 114 farms; all samples were tested using bacterial culture. In addition, a questionnaire was administered to 114 herd owners. Sixty-two percent of the dairy herds had at least 1 positive fecal pool or environmental sample. A total of 218 rabbit samples were collected from 90% of the herds, and 309 deer samples were collected from 47% of the herds. On 2 (4%) of the farms sampled, 1 deer fecal sample was MAP positive. Both farms had samples from the cow fecal pool and cow environment that were positive by culture. On 2 (2%) other farms, 1 rabbit fecal sample was positive by culture to MAP, with one of these farms having positive cow fecal pools and cow environmental samples. Pasture was used on 79% of the study farms as a grazing area for cattle, mainly for dry cows (75%) and bred or prebred heifers (87%). Of the 114 farms, 88 (77%) provided access to drylot for their cattle, mainly for milking cows (77/88; 88%) and bred heifers (87%). Of all study farms, 90 (79%) used some solid manure broadcasting on their crop fields. Of all 114 farms, the estimated probability of daily physical contact between cattle manure and deer or rabbits was 20% and 25%, respectively. Possible contact between cattle manure and deer or rabbits was estimated to occur primarily from March through December. The frequency of pasture or drylot use and manure spreading on crop fields may be important risk factors for transmission of MAP among dairy cattle, deer, and rabbits. Although the MAP prevalence among rabbits and deer is low, their role as MAP reservoirs should be considered. PMID:15745220

Forest soil chemistry and terrain attributes in a Catskills watershed

USGS Publications Warehouse

Johnson, C.E.; Ruiz-Mendez, J. J.; Lawrence, G.B.

2000-01-01

Knowledge of soil chemistry is useful in assessing the sensitivity of forested areas to natural and anthropogenic disturbances, but characterizing large areas is expensive because of the large sample numbers required and the cost of soil chemical analyses. We collected and chemically analyzed soil samples from 72 sites within a 214-ha watershed in the Catskill Mountains of New York to evaluate factors that influence soil chemistry and whether terrain features could be used to predict soil chemical properties. Using geographic information system (GIS) techniques, we determined five terrain attributes at each sampling location: (i) slope, (ii) aspect, (iii) elevation, (iv) topographic index, and (v) flow accumulation. These attributes were ineffective in predicting the chemical properties of organic and mineral soil samples; together they explained only 4 to 25% of the variance in pH(w), effective cation-exchange capacity (CEC(e)), exchangeable bases, exchangeable acidity, total C, total N, and C/N ratio. Regressions among soil properties were much better; total C and pH(w) together explained 33 to 66% of the variation in exchangeable bases and CEC(e). Total C was positively correlated with N (r = 0.91 and 0.96 in Oa horizons and mineral soil, respectively), exchangeable bases (r = 0.65, 0.76), and CEC(e) (r = 0.54, 0.44), indicating the importance of organic matter to the chemistry of these acidic soils. The fraction of CEC(e) occupied by H explained 44% of the variation in pH(w). Soil chemical properties at this site vary on spatial scales finer than typical GIS analyses, resulting in relationships with poor predictive power. Thus, interrelationships among soil properties are more reliable for prediction.Knowledge of soil chemistry is useful in assessing the sensitivity of forested areas to natural and anthropogenic disturbances, but characterizing large areas is expensive because of the large sample numbers required and the cost of soil chemical analyses. We collected and chemically analyzed soil samples from 72 sites within a 214-ha watershed in the Catskill Mountains of New York to evaluate factors that influence soil chemistry and whether terrain features could be used to predict soil chemical properties. Using geographic information system (GIS) techniques, we determined five terrain attributes at each sampling location: (i) slope, (ii) aspect, (iii) elevation, (iv) topographic index, and (v) flow accumulation. These attributes were ineffective in predicting the chemical properties of organic and mineral soil samples; together they explained only 4 to 25% of the variance in pHw, effective cation-exchange capacity (CECe), exchangeable bases, exchangeable acidity, total C, total N, and C/N ratio. Regressions among soil properties were much better; total C and pHw together explained 33 to 66% of the variation in exchangeable bases and CECe. Total C was positively correlated with N (r = 0.91 and 0.96 in Oa horizons and mineral soil, respectively), exchangeable bases (r = 0.65, 0.76), and CECe (r = 0.54, 0.44), indicating the importance of organic matter to the chemistry of these acidic soils. The fraction of CECe occupied by H explained 44% of the variation in pHw. Soil chemical properties at this site vary on spatial scales finer than typical GIS analyses, resulting in relationships with poor predictive power. Thus, interrelationships among soil properties are more reliable for prediction.

[Investigation on sanitation of freshwater aquaculture environments and Clonorchis sinensis intermediate host infection in a city of Pearl River Delta region, China].

PubMed

Man, Wang; Le, Luo; Xue-Qin, Chen; Lei, Li; Yue-Yi, Fang

2017-10-19

To understand the current status of the sanitation of freshwater aquaculture environments, and Clonorchis sinensis infection of freshwater fish in the aquaculture and market in a city of Pearl River Delta region, so as to provide the evidence for formulating the prevention and control strategy of clonorchiasis sinensis. In 2016, based on the distribution of freshwater aquaculture, 36 freshwater fish ponds among 14 towns were selected for sampling and investigation, and 10-20 pieces were collected from each pond. Besides, 3 aquatic product wholesale markets were included, among which 3-6 stalls were selected from each market, and 20-30 pieces were collected from each stall. The metacercaria in the fish was examined by the digestion method. In the 36 fish ponds, there were no toilets with the stool being drained into fish ponds directly, and there was only one pond with duck sheds with the stool being drained into fish ponds directly. Totally 437 pieces of freshwater fish from ponds were detected, with a metacercaria positive rate of 4.35% (19/437). The metacercaria positive fish were distributed in 50% (7/14) of towns and 25% (9/36) ponds. The positive rates of crucian carp, grass carp, dace, aristichthysnobilis, and tilapia were 13.95% (6/43), 4.76% (9/189), 4.44 (2/45), 1.55% (2/129), and 0 (0/31) respectively, with statistically significant difference ( χ 2 = 13.46, P = 0.01). Totally 307 pieces of freshwater fish were collected from the wholesale markets, with a total positive rate of 1.95% (6/307). The positive rate of grass carp and aristichthysnobilis were 3.20% (4/125) and 2.78% (2/72) respectively, and no positive samples were found in crucian carp, dace and tilapia, with no statistically significant difference among the different fish in the infection rate (Fisher exact P = 0.75). The sanitation of freshwater aquaculture environments in a city of Pearl River Delta region is relative good. However, there are different degrees of Clonorchis sinensis infection of freshwater fish in some aquaculture environments and markets.

Molecular diagnosis of Toxoplasma gondii infection in Libya.

PubMed

Gashout, Aisha; Amro, Ahmad; Erhuma, Mabruk; Al-Dwibe, Hamida; Elmaihub, Eanas; Babba, Hamouda; Nattah, Nabil; Abudher, Abdalhafid

2016-04-16

Toxoplasma gondii infections are prevalent in humans and animals throughout Libya. Current diagnosis is based on detection of Toxoplasma-specific IgM and IgG. In this study, we established and optimized a diagnostic PCR assay for molecular diagnosis of T. gondii in Libya. From January to December, 2010, 177 blood and serum samples were collected from suspected patients. This includes: 140 women who have had spontaneous abortions, 26 HIV-positive patients, nine patients with leukemia and lymphoma, and two infants with ocular infection. Samples were screened for anti-Toxoplasma IgG and IgM antibodies before DNA extraction. The surface antigen gene 2 (SAG2) was targeted in a semi-nested PCR to amplify a 999 bp and a 614 bp fragment in the first and the second run respectively. A total of 54/140 (38.5 %) women who have had spontaneous abortions, 23/26 (88 %) HIV patients, 6/9 (66.6 %) of the leukaemia and lymphoma patients, and one child with ocular infection were seropositive for anti-Toxoplasma IgG and/or IgM. Genomic DNA was extracted from 38 selected seropositive samples. The PCR was sensitive enough to detect DNA concentration of 12 ng/μL. PCR analysis was performed for 38 selected seropositive patients (16 women who have had spontaneous abortions, 15 positive HIV patients, six leukaemia patients and one child with ocular infection). Our designed primers were successfully amplified in 22/38 (57.9 %) samples; 5/12 (35.7 %) from serum and 17/26 (65.8 %) from whole blood samples. All PCR positive samples were IgG-positive except two samples which were IgM and IgG & IgM-positive serum samples respectively. The semi-nested PCR confirmed five more samples. These included two leukaemia and two HIV-positive whole blood samples and one serum sample from an aborted woman. The ability of PCR to diagnose active toxoplasmosis is needed in immunocompromised patients and congenital toxoplasmosis cases, especially when serological techniques fail. For the first time in Libya, we established and optimized semi-nested PCR of SAG2 gene. The developed PCR method was able to detect as little as 12 ng/μL of T. gondii DNA and was useful to diagnose the diseases in women who have had spontaneous abortions, HIV-positive patients, patients with leukemia and lymphoma, and infants with ocular infection.

[Detection of Cryptospordium spp. in environmental water samples by FTA-PCR].

PubMed

Zhang, Xiao-Ping; Zhu, Qian; He, Yan-Yan; Jiang, Li; Jiang, Shou-Fu

2011-02-01

To establish a FTA-polymeras chain reaction (FTA-PCR) method in detection of Cryptospordium spp. in different sources of water. The semi automated immunomagnetic separation (IMS) of Cryptospordium oocysts in environmental water samples was performed firstly, and then genomic DNA of Cryptospordium oocysts was extracted by FTA filters disk. Oligonucleotide primers were designed based on the DNA fragment of the 18 S rRNA gene from C. parvum. Plate DNA was amplified with primers in PCR. The control DNA samples from Toxoplasma gondii,Sarcocystis suihominis, Echinococcus granulosus, and Clonorchis sinensis were amplified simultaneously. All PCR products were detected by agar electrophoresis dyed with ethidium bromide. The 446 bp fragment of DNA was detected in all samples of C. parvum, C. andersoni, and C. baileyi, while it was not detected in control groups in laboratory. No positive samples were found from 10 samples collected from tape water in 5 districts of Shanghai City by FTA-PCR. Nine positive samples were detected totally from 70 different environmental water samples, there were 0 out of 15 samples from the source of tape water, 2 out of 25 from the Huangpu River, 5 out of 15 from rivers around the animal farmers, 1 out of 9 from output water of contaminating water treatment factory, 1 out of 6 from the out gate of living contaminating water. The 446 bp fragment was detected from all the amplified positive water samples. FTA-PCR is an efficient method for gene detection of Cryptospordium oocysts, which could be used in detection of environmental water samples. The contamination degree of Cryptospordium oocysts in the river water around animal farms is high.

Detection of autoantibodies against thyroid peroxidase in serum samples of hypothyroid dogs.

PubMed

Skopek, Evelyn; Patzl, Martina; Nachreiner, Raymond F

2006-05-01

To establish a sensitive test for the detection of autoantibodies against thyroid peroxidase (TPO) in canine serum samples. 365 serum samples from dogs with hypothyroidism as determined on the basis of serum concentrations of total and free triiodothyronine (T3), total and free thyroxine (T4), and thyroid-stimulating hormone, of which 195 (53%) had positive results for at least 1 of 3 thyroid autoantibodies (against thyroglobulin [Tg], T4, or T3) and serum samples from 28 healthy dogs (control samples). TPO was purified from canine thyroid glands by extraction with detergents, ultracentrifugation, and precipitation with ammonium sulfate. Screening for anti-TPO autoantibodies in canine sera was performed by use of an immunoblot assay. Thyroid extract containing TPO was separated electrophoretically, blotted, and probed with canine sera. Alkaline phosphatase-conjugated rabbit anti-dog IgG was used for detection of bound antibodies. TPO bands were observed at 110, 100, and 40 kd. Anti-TPO autoantibodies against the 40-kd fragment were detected in 33 (17%) sera of dogs with positive results for anti-Tg, anti-T4, or anti-T3 autoantibodies but not in sera of hypothyroid dogs without these autoantibodies or in sera of healthy dogs. The immunoblot assay was a sensitive and specific method for the detection of autoantibodies because it also provided information about the antigen. Anti-TPO autoantibodies were clearly detected in a fraction of hypothyroid dogs. The value of anti-TPO autoantibodies for use in early diagnosis of animals with thyroid gland diseases should be evaluated in additional studies.

Effect of intertidal exposure on Vibrio parahaemolyticus levels in Pacific Northwest oysters.

PubMed

Nordstrom, J L; Kaysner, C A; Blackstone, G M; Vickery, M C L; Bowers, J C; DePaola, A

2004-10-01

Interest in Vibrio parahaemolyticus (Vp) increased in the United States following Vp-associated gastroenteritis outbreaks in 1997 and 1998 involving the West Coast and other areas. The present study evaluated multiple aspects of Vp ecology in the Pacific Northwest with three objectives: (i) to determine the effect of low-tide exposure on Vp levels in oysters, (ii) to determine the relationship between total and pathogenic Vp, and (iii) to examine sediments and aquatic fauna as reservoirs for pathogenic Vp. Samples were collected from intertidal reefs along Hood Canal, Wash., in August 2001. Fecal matter from marine mammals and aquatic birds as well as intestinal contents from bottom-dwelling fish were tested. Total and pathogenic Vp levels in all the samples were enumerated with colony hybridization procedures using DNA probes that targeted the thermolabile direct hemolysin (tlh) and thermostable direct hemolysin (tdh) genes, respectively. The mean Vp densities in oysters were four to eight times greater at maximum exposure than at the corresponding first exposure. While tdh-positive Vp counts were generally < or = 10 CFU/g at first exposure, counts as high as 160 CFU/g were found at maximum exposure. Vp concentrations in sediments were not significantly different from those in oysters at maximum exposure. Pathogenic (tdh positive) Vp was detected in 9 of 42 (21%) oyster samples at maximum exposure, in 5 of 19 (26%) sediment samples, but in 0 of 9 excreta samples. These results demonstrate that summer conditions permit the multiplication of Vp in oysters exposed by a receding tide.

Antioxidant properties, physico-chemical characteristics and proximate composition of five wild fruits of Manipur, India.

PubMed

Sharma, Ph Baleshwor; Handique, Pratap Jyoti; Devi, Huidrom Sunitibala

2015-02-01

Antioxidant properties, physico-chemical characteristics and proximate composition of five wild fruits viz., Garcinia pedunculata, Garcinia xanthochymus, Docynia indica, Rhus semialata and Averrhoa carambola grown in Manipur, India were presented in the current study. The order of the antioxidant activity and reducing power of the fruit samples was found as R. semialata > D. indica > G. xanthochymus > A. carambola > G. pedunculata. Good correlation coefficient (R(2) > 0.99) was found among the three methods applied to determine antioxidant activity. Total phenolic content was positively correlated (R(2) = 0.960) with the antioxidant activity however, total flavonoid content was not positively correlated with the antioxidant activity. Physico-chemical and proximate composition of these fruits is documented for the first time.

Evaluation of negative results of BacT/Alert 3D automated blood culture system.

PubMed

Kocoglu, M Esra; Bayram, Aysen; Balci, Iclal

2005-06-01

Although automated continuous-monitoring blood culture systems are both rapid and sensitive, false-positive and false-negative results still occur. The objective of this study, then, was to evaluate negative results occurring with BacT/Alert 3D blood culture systems. A total of 1032 samples were cultured with the BacT/Alert 3D automated blood culture system, using both aerobic (FA) and anaerobic (FN) [corrected] media, and 128 of these samples yielded positive results. A total of 904 negative blood samples were then subcultured in 5% sheep blood agar, eosin methylene blue, chocolate agar, and sabouraud-dextrose agar. Organisms growing on these subcultures were subsequently identified using both Vitek32 (bioMerieux, Durham, NC) and conventional methods. Twenty four (2.6%) of the 904 subcultures grew on the subculture media. The majority (83.3%) of these were determined to be gram-positive microorganisms. Fourteen (58.3%) were coagulase-negative staphylococci, two (8.3%) were Bacillus spp., one (4.2%) was Staphylococcus aureus, and one (4.2%) was identified as Enterococcus faecium. Streptococcus pneumoniae and Neisseria spp. were isolated together in two (8.3%) vials. Gram-negative microorganisms comprised 12.5% of the subcultures, of which two (8.3%) were found to be Pseudomonas aeruginosa, and one (4.2%) was Pseudomonas fluorescens. The other isolate (4.2%) was identified as Candida albicans. We conclude that the subculture of negative results is valuable in the BacT/Alert 3D system, especially in situations in which only one set of blood cultures is taken.

Gratitude mediates the effect of emotional intelligence on subjective well-being: A structural equation modeling analysis.

PubMed

Geng, Yuan

2016-11-01

This study investigated the relationship among emotional intelligence, gratitude, and subjective well-being in a sample of university students. A total of 365 undergraduates completed the emotional intelligence scale, the gratitude questionnaire, and the subjective well-being measures. The results of the structural equation model showed that emotional intelligence is positively associated with gratitude and subjective well-being, that gratitude is positively associated with subjective well-being, and that gratitude partially mediates the positive relationship between emotional intelligence and subjective well-being. Bootstrap test results also revealed that emotional intelligence has a significant indirect effect on subjective well-being through gratitude.

Comparing the biocidal properties of non-thermal plasma sources by reference protocol

NASA Astrophysics Data System (ADS)

Khun, Josef; Jirešová, Jana; Kujalová, Lucie; Hozák, Pavel; Scholtz, Vladimír

2017-10-01

The previously proposed reference protocol enabling easy comparison of biocidal properties of different non-thermal plasma sources has been followed and discussed. For inactivation tests the reference protocol has used spores of Gram positive bacterium Bacillus subtilis (ATCC 6633) deposited on a polycarbonate membrane as reference sample. In this work, biocidal properties of a negative glow corona, positive streamer corona, positive transient spark and cometary discharges are being compared in both open air and closed apparatus. Despite the total number of bacteria surviving 1 h exposure has decreased by up to 7 orders in closed apparatus, in open one, only weak inhibition bactericidal effect has been observed.

Carbapenem-Resistant Acinetobacter baumannii: Concomitant Contamination of Air and Environmental Surfaces.

PubMed

Shimose, Luis A; Masuda, Eriko; Sfeir, Maroun; Berbel Caban, Ana; Bueno, Maria X; dePascale, Dennise; Spychala, Caressa N; Cleary, Timothy; Namias, Nicholas; Kett, Daniel H; Doi, Yohei; Munoz-Price, L Silvia

2016-07-01

OBJECTIVE To concomitantly determine the differential degrees of air and environmental contamination by Acinetobacter baumannii based on anatomic source of colonization and type of ICU layout (single-occupancy vs open layout). DESIGN Longitudinal prospective surveillance study of air and environmental surfaces in patient rooms. SETTING A 1,500-bed public teaching hospital in Miami, Florida. PATIENTS Consecutive A. baumannii-colonized patients admitted to our ICUs between October 2013 and February 2014. METHODS Air and environmental surfaces of the rooms of A. baumannii-colonized patients were sampled daily for up to 10 days. Pulsed-field gel electrophoresis (PFGE) was used to type and match the matching air, environmental, and clinical A. baumannii isolates. RESULTS A total of 25 A. baumannii-colonized patients were identified during the study period; 17 were colonized in the respiratory tract and 8 were colonized in the rectum. In rooms with rectally colonized patients, 38.3% of air samples were positive for A. baumannii; in rooms of patients with respiratory colonization, 13.1% of air samples were positive (P=.0001). In rooms with rectally colonized patients, 15.5% of environmental samples were positive for A. baumannii; in rooms of patients with respiratory colonization, 9.5% of environmental samples were positive (P=.02). The rates of air contamination in the open-layout and single-occupancy ICUs were 17.9% and 21.8%, respectively (P=.5). Environmental surfaces were positive in 9.5% of instances in open-layout ICUs versus 13.4% in single-occupancy ICUs (P=.09). CONCLUSIONS Air and environmental surface contaminations were significantly greater among rectally colonized patients; however, ICU layout did not influence the rate of contamination. Infect Control Hosp Epidemiol 2016;37:777-781.

Multiplex Real-Time PCR Method for Simultaneous Identification and Toxigenic Type Characterization of Clostridium difficile From Stool Samples

PubMed Central

Alam, Mohammad J.; Tisdel, Naradah L.; Shah, Dhara N.; Yapar, Mehmet; Lasco, Todd M.; Garey, Kevin W.

2015-01-01

Background The aim of this study was to develop and validate a multiplex real-time PCR assay for simultaneous identification and toxigenic type characterization of Clostridium difficile. Methods The multiplex real-time PCR assay targeted and simultaneously detected triose phosphate isomerase (tpi) and binary toxin (cdtA) genes, and toxin A (tcdA) and B (tcdB) genes in the first and sec tubes, respectively. The results of multiplex real-time PCR were compared to those of the BD GeneOhm Cdiff assay, targeting the tcdB gene alone. The toxigenic culture was used as the reference, where toxin genes were detected by multiplex real-time PCR. Results A total of 351 stool samples from consecutive patients were included in the study. Fifty-five stool samples (15.6%) were determined to be positive for the presence of C. difficile by using multiplex real-time PCR. Of these, 48 (87.2%) were toxigenic (46 tcdA and tcdB-positive, two positive for only tcdB) and 11 (22.9%) were cdtA-positive. The sensitivity, specificity, negative predictive value (NPV), and positive predictive value (PPV) of the multiplex real-time PCR compared with the toxigenic culture were 95.6%, 98.6%, 91.6%, and 99.3%, respectively. The analytical sensitivity of the multiplex real-time PCR assay was determined to be 103colonyforming unit (CFU)/g spiked stool sample and 0.0625 pg genomic DNA from culture. Analytical specificity determined by using 15 enteric and non-clostridial reference strains was 100%. Conclusions The multiplex real-time PCR assay accurately detected C. difficile isolates from diarrheal stool samples and characterized its toxin genes in a single PCR run. PMID:25932438

Analysis of student attitudes towards e-learning using Fishbein Multiattribute approach

NASA Astrophysics Data System (ADS)

Jasuli

2018-01-01

This research aimed to know students’ attitudes toward e-learning and to determine what attributes were considered to be dominant by students toward the use of e-learning. The research population was all postgraduate students in the 2016 academic year at Universitas Negeri Surabaya. The sampling technique is using nonprobability sampling and purposive sampling with the sample totaled 100 respondents. The research instrument is using questionnaire with semantic differential scale. The models used to analyze is multi-attribute Fishbein model. The findings indicated that student attitudes toward e-learning are positive and easy accessibility which is considered as the most important attribute by students toward the use of e-learning.

Quantitative Polymerase Chain Reaction to Assess Response to Treatment of Bacterial Vaginosis and Risk of Preterm Birth.

PubMed

Abramovici, Adi; Lobashevsky, Elena; Cliver, Suzanne P; Edwards, Rodney K; Hauth, John C; Biggio, Joseph R

2015-10-01

The aim of this study was to determine whether quantitative polymerase chain reaction (qPCR) bacterial load measurement is a valid method to assess response to treatment of bacterial vaginosis and risk of preterm birth in pregnant women. Secondary analysis by utilizing stored vaginal samples obtained during a previous randomized controlled trial studying the effect of antibiotics on preterm birth (PTB). All women had risk factors for PTB: (1) positive fetal fibronectin (n=146), (2) bacterial vaginosis (BV) and a prior PTB (n=43), or (3) BV and a prepregnancy weight<50 kg (n=54). Total and several individual BV-related bacteria loads were measured using qPCR for 16S rRNA. Loads were correlated with Nugent scores (Spearman correlation coefficients). Loads were compared pre- and posttreatment with Wilcoxon rank-sum test. Individual patient differences were examined with Wilcoxon signed-rank test. A total of 243 paired vaginal samples were available for analysis: 123 antibiotics and 120 placebo. Groups did not differ by risk factors for PTB. For all samples, bacterial loads were correlated with Nugent score and each of its specific bacterial components (all p<0.01). Baseline total bacterial load did not differ by treatment group (p=0.87). Posttreatment total bacterial load was significantly lower in the antibiotics group than the placebo group (p<0.01). Individual patient total bacterial load decreased significantly posttreatment in the antibiotics group (p<0.01), but not in the placebo group (p=0.12). The rate of PTB did not differ between groups (p=0.24). PTB relative risks calculated for BV positive versus BV negative women and women with the highest quartile total and individual bacterial loads were not statistically significant. qPCR correlates with Nugent score and demonstrates decreased bacterial load after antibiotic treatment. Therefore, it is a valid method of vaginal flora assessment in pregnant women who are at high risk for PTB. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

Association between Macronutrients Intake, Visceral Obesity and Blood Pressure in a Sample of Obese Egyptian Women.

PubMed

Hassan, Nayera E; El Shebini, Salwa M; Ahmed, Nihad H; Selim Mostafa, Mohamed

2015-03-15

Study the association between the total caloric intake, protein, lipid, and some classes of fatty acids of the diet, and their effects on blood pressure in a sample of Egyptian obese women with and without visceral obesity. Five hundred forty-nine obese women were included in the study with mean age of 38.1 ± 11.56 years and mean Body mass index [BMI] of 36.17 ± 7.23. They enrolled in a program for losing weight. Visceral fat was determined using ultrasound. Blood pressure was measured 3 times and the mean was recorded. Twenty four hours dietary recall was reported. Thirty point four percentages of samples has visceral obesity ≥ 7cm; they were the older, showed higher values of BMI, visceral obesity and blood pressure. Significant difference was found between groups regarding mean value of BMI, visceral obesity, both systolic blood pressure SBP and diastolic blood pressure DBP and most of the daily macronutrients intake. In groups (2&3) positive significant correlation was recorded between (SBP) & (DBP) and total daily intake of total calories, carbohydrate, total fat, saturated fatty acids and cholesterol, and negative significant correlation with total daily intake of total protein, animal and vegetable protein, linolenic and linoleic fatty acids, while oleic fatty acid showed negative correlation with SBP&DBP in all groups. This study emphasizes the hypothesis that the macronutrients composition of diet influences blood pressure in different ways, in obese patients with visceral obesity.

Association between Macronutrients Intake, Visceral Obesity and Blood Pressure in a Sample of Obese Egyptian Women

PubMed Central

Hassan, Nayera E.; El Shebini, Salwa M.; Ahmed, Nihad H.; Selim Mostafa, Mohamed

2015-01-01

AIM: Study the association between the total caloric intake, protein, lipid, and some classes of fatty acids of the diet, and their effects on blood pressure in a sample of Egyptian obese women with and without visceral obesity. METHODS: Five hundred forty-nine obese women were included in the study with mean age of 38.1 ± 11.56 years and mean Body mass index [BMI] of 36.17 ± 7.23. They enrolled in a program for losing weight. Visceral fat was determined using ultrasound. Blood pressure was measured 3 times and the mean was recorded. Twenty four hours dietary recall was reported. RESULTS: Thirty point four percentages of samples has visceral obesity ≥ 7cm; they were the older, showed higher values of BMI, visceral obesity and blood pressure. Significant difference was found between groups regarding mean value of BMI, visceral obesity, both systolic blood pressure SBP and diastolic blood pressure DBP and most of the daily macronutrients intake. In groups (2&3) positive significant correlation was recorded between (SBP) & (DBP) and total daily intake of total calories, carbohydrate, total fat, saturated fatty acids and cholesterol, and negative significant correlation with total daily intake of total protein, animal and vegetable protein, linolenic and linoleic fatty acids, while oleic fatty acid showed negative correlation with SBP&DBP in all groups. CONCLUSION: This study emphasizes the hypothesis that the macronutrients composition of diet influences blood pressure in different ways, in obese patients with visceral obesity. PMID:27275219

Sample pooling for real-time PCR detection and virulence determination of the footrot pathogen Dichelobacter nodosus.

PubMed

Frosth, Sara; König, Ulrika; Nyman, Ann-Kristin; Aspán, Anna

2017-09-01

Dichelobacter nodosus is the principal cause of ovine footrot and strain virulence is an important factor in disease severity. Therefore, detection and virulence determination of D. nodosus is important for proper diagnosis of the disease. Today this is possible by real-time PCR analysis. Analysis of large numbers of samples is costly and laborious; therefore, pooling of individual samples is common in surveillance programs. However, pooling can reduce the sensitivity of the method. The aim of this study was to develop a pooling method for real-time PCR analysis that would allow sensitive detection and simultaneous virulence determination of D. nodosus. A total of 225 sheep from 17 flocks were sampled using ESwabs within the Swedish Footrot Control Program in 2014. Samples were first analysed individually and then in pools of five by real-time PCR assays targeting the 16S rRNA and aprV2/B2 genes of D. nodosus. Each pool consisted of four negative and one positive D. nodosus samples with varying amounts of the bacterium. In the individual analysis, 61 (27.1%) samples were positive in the 16S rRNA and the aprV2/B2 PCR assays and 164 (72.9%) samples were negative. All samples positive in the aprV2/B2 PCR-assay were of aprB2 variant. The pooled analysis showed that all 41 pools were also positive for D. nodosus 16S rRNA and the aprB2 variant. The diagnostic sensitivity for pooled and individual samples was therefore similar. Our method includes concentration of the bacteria before DNA-extraction. This may account for the maintenance of diagnostic sensitivity. Diagnostic sensitivity in the real-time PCR assays of the pooled samples were comparable to the sensitivity obtained for individually analysed samples. Even sub-clinical infections were able to be detected in the pooled PCR samples which is important for control of the disease. This method may therefore be implemented in footrot control programs where it can replace analysis of individual samples.

Serological, molecular detection and potential risk factors associated with camel brucellosis in Pakistan.

PubMed

Fatima, Sana; Khan, Iahtasham; Nasir, Amar; Younus, Muhammad; Saqib, Muhammad; Melzer, Falk; Neubauer, Heinrich; El-Adawy, Hosny

2016-12-01

Brucellosis is one of the most important zoonoses in developing countries and was considered the most widespread zoonosis in the world. Brucellosis was reported in camels and has been reported from all camel-keeping countries.The present study was performed in three districts (Jhang, Chiniot, and Bhakkar) of Punjab province of Pakistan. A total of 200 camel (Camelus bactrianus) sera were collected using random and multistage cluster sampling from different areas. Fifty samples were collected from one organized governmental farm. One hundred fifty samples were collected randomly from nomadic/pastoral production systems. All sera were tested with Rose Bengal plate agglutination test (RBPT) and confirmed by ELISA. Genomic DNA was extracted from all serum samples and tested by real-time PCR. Various potential risk factors (season, rearing with other animals, and abortion or orchitis history) recorded through questionnaires were statistically analyzed by Chi-square test.In total, 5 % of investigated sera were positive by RBPT. Only 2 % of the camel sera were CELISA positive. Brucella abortus DNA was detected in 1.5 % of the investigated animals. Season, rearing of camels with other ruminants, abortion, and orchitis history were found to be statistically significant (p < 0.05) disease for determinants.Camel brucellosis is a zoonotic disease in the Pakistani Punjab with various risk factors maintaining and perpetuating its spread. Therefore, there is a need for implementing control measures and raising public health awareness in prevention of brucellosis in Pakistan.

SEROPREVALENCE AND GENETIC CHARACTERIZATION OF TOXOPLASMA GONDII IN NATURALLY-INFECTED SYNANTHROPIC RODENTS IN YUNNAN PROVINCE, SOUTHWESTERN CHINA.

PubMed

Wang, Xin-Lei; Dong, Ling; Zhang, Li; Lv, Yan; Li, Qian; Li, Hailong

2018-05-03

Rodents play important rule in the transmission of Toxoplasma gondii, however, little is known of the seroprevalence and genetic diversity of T. gondii infection in rodents in Yunnan, southwestern China. In this study, we examined the seroprevalence and genetically characterized T. gondii infection in naturally-infected rodents in Yunnan Province, southwestern China between March and July 2016. A total of 261 serum samples were collected from rodents in Jingha district, Xishuangbanna Dai Autonomous Prefecture, Yunnan province, and examined for T. gondii antibodies by modified agglutination test (MAT) at a 1:20 cut-off, with titers of 1:20 in 24, 1:40 in 10, 1:80 in 10, 1:160 in 4. Thirty-two of 261 (12.26%) serum samples were positive for T. gondii by MAT. A total of 751 rodent brain samples were examined by semi-nested-PCR, 46 (6.13%) were positive for the T. gondii B1 gene, 2 of them showed complete genotyping results for all 11 polymorphic loci (SAG1, SAG2, alt.SAG2, SAG3, BTUB, GRA6, L358, PK1, C22-8, C29-2 and Apico). The two samples were characterized to represent ToxoDB Genotype #137 (http://toxodb.org/toxo/). These findings enriched the epidemiological and genetic diversity of T. gondii in China, and implications for the better prevention and control of T. gondii infection in humans and animals.

DETECTION OF CANINE PARVOVIRUS ANTIGEN IN DOGS IN KUMASI, GHANA

PubMed Central

Folitse, R. D; Kodie, D.O; Amemor, E.; Dei, D.; Tasiame, W.; Burimuah, V.; Emikpe, B.O

2018-01-01

Background: Canine Parvovirus (CPV) in dogs has been documented in many countries. However, evidence of the infection is scanty in Ghana. This study was conducted to detect canine parvovirus antigen in dogs presented with diarrhoea to the Government Veterinary Clinic in Kumasi, Ghana. Materials and Methods: Faecal samples from 72 dogs presented with diarrhoea were tested for the presence of canine parvovirus antigen using commercially available rapid test kit (BIT® Rapid Colour Canine Parvovirus Ag Test Kit, BIOINDIST Co. Ltd, Korea) based on the principle of immunochromatography. Influence of breed, sex, age, vaccination history and the nature of diarrhoea were assessed. Data obtained was analysed with SPSS and subjected to the chi-square test. Significance was at α0.05 Results: We found 61.11% tested positive (44/72) for CPV. Based on sex, 61.54% of males (20/33) and 60.61% of females tested positive (24/39). A total of 65.67% of samples from puppies below 6 months were positive. 56.25% of CPV vaccinated dogs and 70.83% of unvaccinated dogs were positive respectively. 69.05% of samples from haemorrhagic diarrhoeic dogs and 50.00% from non-haemorrhagic diarrhoeic dogs were positive of CPV. Conclusion: The study is the first documented evidence of the existence of CPV in Ghana. It also revealed that absence of bloody diarrhoea does not necessarily rule out CPV infection. PMID:29302647

DETECTION OF CANINE PARVOVIRUS ANTIGEN IN DOGS IN KUMASI, GHANA.

PubMed

Folitse, R D; Kodie, D O; Amemor, E; Dei, D; Tasiame, W; Burimuah, V; Emikpe, B O

2018-01-01

Canine Parvovirus (CPV) in dogs has been documented in many countries. However, evidence of the infection is scanty in Ghana. This study was conducted to detect canine parvovirus antigen in dogs presented with diarrhoea to the Government Veterinary Clinic in Kumasi, Ghana. Faecal samples from 72 dogs presented with diarrhoea were tested for the presence of canine parvovirus antigen using commercially available rapid test kit (BIT ® Rapid Colour Canine Parvovirus Ag Test Kit, BIOINDIST Co. Ltd, Korea) based on the principle of immunochromatography. Influence of breed, sex, age, vaccination history and the nature of diarrhoea were assessed. Data obtained was analysed with SPSS and subjected to the chi-square test. Significance was at α 0.05 . We found 61.11% tested positive (44/72) for CPV. Based on sex, 61.54% of males (20/33) and 60.61% of females tested positive (24/39). A total of 65.67% of samples from puppies below 6 months were positive. 56.25% of CPV vaccinated dogs and 70.83% of unvaccinated dogs were positive respectively. 69.05% of samples from haemorrhagic diarrhoeic dogs and 50.00% from non-haemorrhagic diarrhoeic dogs were positive of CPV. The study is the first documented evidence of the existence of CPV in Ghana. It also revealed that absence of bloody diarrhoea does not necessarily rule out CPV infection.

The unexpected discovery of Brucella abortus Buck 19 vaccine in goats from Ecuador underlines the importance of biosecurity measures.

PubMed

Ron-Román, Jorge; Berkvens, Dirk; Barzallo-Rivadeneira, Daniela; Angulo-Cruz, Alexandra; González-Andrade, Pablo; Minda-Aluisa, Elizabeth; Benítez-Ortíz, Washington; Brandt, Jef; Rodríguez-Hidalgo, Richar; Saegerman, Claude

2017-03-01

Very few, mostly old, and only preliminary serological studies of brucellosis in goats exist in Ecuador. In order to assess the current epidemiological situation, we performed a cross-sectional serological study in the goat populations of Carchi (n = 160 animals), Pichincha (n = 224 animals), and Loja provinces (n = 2024 animals). Only two positive serological results (RB negative and SAT-EDTA ≥400 IU/ml) were obtained in lactating goats from the same farm in Quito (Pichincha province). Additionally, milk was sampled from 220 animals in Pichincha province. The present study indicates a low apparent prevalence in Pichincha province and absence in Carchi and Loja provinces. A total of 25 positive milk ring tests (MRT) were obtained in Pichincha province yielding a prevalence of MRT of 11.16%. Subsequent culture was performed on the positive MRT samples. All results were negative, apart from a single sample, obtained from a serologically positive goat in Quito, that was positive for Brucella abortus strain 19 (B19). Several hypotheses are forwarded concerning this unexpected result. The most likely hypothesis is the possible accidental use of a needle, previously used for vaccination of cattle with the said vaccine, for the administration of drug treatment to the goat. This hypothesis underlines the necessity of biosecurity measures to prevent this type of accidents.

Molecular characterization of Orientia tsutsugamushi serotypes causing scrub typhus outbreak in southern region of Andhra Pradesh, India.

PubMed

Usha, K; Kumar, E; Kalawat, Usha; Kumar, B Siddhartha; Chaudhury, A; Gopal, D V R Sai

2016-10-01

Scrub typhus is a vector-borne zoonotic infection caused by Orientiatsutsugamushi. Local epidemiology of the circulating serotypes of scrub typhus is not available from most parts of India. We conducted this study for the diagnosis of scrub typhus using IgM ELISA and to detect O. tsutsugamushi serotypes circulating in southern Andhra Pradesh, India. Samples were collected from patients clinically suspected to have scrub typhus and were subjected to IgM ELISA to measure IgM antibodies against O. tsutsugamushi. Nested polymerase chain reaction (PCR) was performed targeting strain-specific regions in ELISA-positive samples. Of a total of 663 samples, 258 (38.91%) were found to be positive by IgM ELISA. Serotypes could be detected in 230 (34.69%) samples only. Only two serotypes, Karp and Kawasaki, were found in the serum samples, with the former being predominant. The dual infection of Karp and Kawasaki serotypes was found in seven patients. Other serotypes such as Gilliam, Kuroki and Kato were not detected in the samples. The nested PCR products proved useful in presumptively identifying the endemic O. tsutsugamushi serotypes. The present study could be significant in understanding scrub typhus epidemiology in this region.

Occurrence, Persistence, and Virulence Potential of Listeria ivanovii in Foods and Food Processing Environments in the Republic of Ireland

PubMed Central

Morgan, Ciara A.; Hill, Colin; Gahan, Cormac G. M.; Jordan, Kieran

2015-01-01

The aim of this study was to assess the occurrence of L. ivanovii in foods and food processing environments in Ireland, to track persistence, and to characterize the disease causing potential of the isolated strains. A total of 2,006 samples (432 food samples and 1,574 environmental swabs) were collected between March 2013 and March 2014 from 48 food business operators (FBOs) belonging to different production sectors (dairy, fish, meat, and fresh-cut vegetable). Six of the forty-eight FBOs had samples positive for L. ivanovii on at least one sampling occasion. L. ivanovii was present in fifteen samples (fourteen environmental samples and one food sample). All but one of those positive samples derived from the dairy sector, where L. ivanovii prevalence was 1.7%. Six distinguishable pulsotypes were obtained by PFGE analysis, with one pulsotype being persistent in the environment of a dairy food business. Sequence analysis of the sigB gene showed that fourteen isolates belonged to L. ivanovii subsp. londoniensis, while only one isolate was L. ivanovii subsp. ivanovii. Cell invasion assays demonstrated that the majority of L. ivanovii strains were comparable to L. monocytogenes EGDe in their ability to invade CACO-2 epithelial cells whilst four isolates had significantly higher invasion efficiencies. PMID:26543856

Epidemiology and Etiology of Sexually Transmitted Infection among Hotel-Based Sex Workers in Dhaka, Bangladesh

PubMed Central

Nessa, Khairun; Waris, Shama-A; Sultan, Zafar; Monira, Shirajum; Hossain, Maqsud; Nahar, Shamsun; Rahman, Habibur; Alam, Mahbub; Baatsen, Pam; Rahman, Motiur

2004-01-01

The prevalence of sexually transmitted infections (STIs) and reproductive tract infections (RTIs) among hotel-based sex workers (HBSWs) in Dhaka, Bangladesh, was studied. A total of 400 HBSWs were enrolled in the study during April to July 2002. Endocervical swabs, high vaginal swabs, and blood samples from 400 HBSWs were examined for Neisseria gonorrhoeae (by culture), Chlamydia trachomatis (by PCR), Trichomonas vaginalis (by microscopy), antibody to Treponema pallidum (by both rapid plasma reagin and Treponema pallidum hemagglutination tests), and antibody to herpes simplex virus type 2 (HSV-2) (by enzyme-linked immunosorbent assay). Sociodemographic information as well as gynecological and obstetric information was collected. Among the HBSWs, 228 women (57%) were symptomatic and 172 (43%) were asymptomatic, 35.8% were positive for N. gonorrhoeae, 43.5% were positive for C. trachomatis, and 4.3% were positive for T. vaginalis. A total of 8.5% had syphilis, 34.5% were positive for HSV-2, and 86.8% were positive for at least one RTI or STI. There was no significant difference between the prevalences of STIs among the symptomatic and asymptomatic HBSWs. These data suggested a high prevalence of STIs, particularly gonorrhea and chlamydia, among HBSWs in Dhaka. PMID:14766825

Prevalence of Cryptosporidium species among HIV positive asymptomatic and symptomatic immigrant population in Kashmir, India

PubMed Central

Masarat, S; Ahmad, F; Chisti, M; Hamid, S; Sofi, B Ahmad

2012-01-01

Background and Objectives Cryptosporidiosis has not been reported as an endemic disease in Kashmir, but high prevalence of Cryptosporidium sp. has been found among asymptomatic (non-diarrheic) HIV positive immigrants in present study. Due to increasing number of HIV positive immigrants in Kashmir, Cryptosporidium may become a public health problem in Kashmir. Materials and Methods A total of 45 stool samples were obtained from symptomatic (diarrheic n = 9) and asymptomatic (non-diarrheic n = 36) patients infected with HIV. The stool samples were concentrated using formalin ethyl acetate concentration technique, stained with modified Kinyoun's cold stain and oocysts were identified by microscopy under 1000 x magnification. It was confirmed by detection of antigens in stool samples by ELISA. Results It was established that all the patients studied were carriers of Cryptosporidium. In present study though 80% of patients were asymptomatic (non-diarrheic) and HIV positive which involved non-Kashmiri army personals and travelers (immigrants) but were carriers of Cryptosporidium and 20% of HIV positive patients were emigrants (local Kashmiri traders) who travelled different states of India were having diarrhea (symptomatic) as well as carrier of Cryptosporidium. Conclusion Though Cryptosporidium infection causes chronic diarrhea but in present study all HIV positive patients screened whether diarrheic or non-diarrheic were positive for Cryptosporidium. To prevent the transmission of Cryptosporidium oocyst in environment and endemic spread of cryptosporidiosis as non-diarrheic HIV positive population may be potential source of infection, obligatory laboratory testing for Cryptosporidium in HIV positive immigrant population like traders and travelers is highly recommended in order to have a better understanding of the cause of spread Cryptosporidium infection in Kashmir. PMID:22783459

THE PREVALENCE OF CANNABIS-INVOLVED DRIVING IN CALIFORNIA

PubMed Central

Johnson, Mark B.; Kelley-Baker, Tara; Voas, Robert B.; Lacey, John H.

2013-01-01

Background Various national surveys suggest that cannabis use is rising nationally, and many States have passed legislation that has potential to increase usage even further. This presents a problem for public roadways, as research suggests that cannabis impairs driving ability. Methods Anonymous oral fluid samples and breath tests were obtained from more than 900 weekend nighttime drivers randomly sampled from six jurisdictions in California. Oral fluid samples were assayed for the presence of Schedule I drugs. Drivers also completed information on self-reported drug use and possession of a medical cannabis permit. Data from the 2007 National Roadside Survey (collected using comparable methods) were used as a comparison. Results Using the 2010 data, a total of 14.4% of weekend nighttime drivers tested positive for illegal drugs, with 8.5% testing positive for delta-9-tetrahydrocannabinol (THC). THC-positive rates varied considerably among jurisdictions, from a low of 4.3% in Fresno to a high of 18.3% in Eureka. A comparison with the 2007 NRS data found an increase in THC-positive drivers in 2010, but no increase in illegal drugs other than cannabis. Drivers who reported having a medical cannabis permit were significantly more likely to test positive for THC. Conclusions Cannabis-involved driving has increased in California since 2007. Nearly 1-in-10 weekend, nighttime drivers tested positive for THC, and in some jurisdictions, the rate was nearly 1-in-5. The possible contribution of cannabis legislation, such as decriminalization and medical cannabis usage, is discussed. PMID:22101027

Rapid identification of pneumococci, enterococci, beta-haemolytic streptococci and S. aureus from positive blood cultures enabling early reports.

PubMed

Larsson, Marie C; Karlsson, Ewa; Woksepp, Hanna; Frölander, Kerstin; Mårtensson, Agneta; Rashed, Foad; Annika, Wistedt; Schön, Thomas; Serrander, Lena

2014-03-19

The aim of this study was to evaluate diagnostic tests in order to introduce a diagnostic strategy to identify the most common gram-positive bacteria (pneumococci, enterococci, β-haemolytic streptococci and S. aureus) found in blood cultures within 6 hours after signalling growth. The tube coagulase test was optimized and several latex agglutination tests were compared and evaluated before a validation period of 11 months was performed on consecutive positive blood culture patient samples from Kalmar County Hospital, Sweden. During the validation period 150 (91%) of a total of 166 gram-positive cocci (119 in clusters, 45 in chains or pairs and 2 undefined morphology) were correctly identified as S. aureus, CoNS, Pneumococci, Enterococci or group A streptococci (GAS), group B streptococci (GBS), group G streptococci (GGS) within 6 hours with a minimal increase in work-load and costs. The remaining samples (9%) were correctly identified during the next day. No samples were incorrectly grouped with this diagnostic strategy and no patient came to risk by early reporting. A simple strategy gives reliable and cost-effective reporting of >90% of the most common gram-positive cocci within 6 hours after a blood cultures become positive. The high specificity of the tests used makes preliminary reports reliable. The reports can be used to indicate the focus of infection and not the least, support faster administration of proper antimicrobial treatment for patients with serious bacterial infections.

Geographic distribution and incidence of Lyme borreliosis in the west of Ireland.

PubMed

Vellinga, A; Kilkelly, H; Cullinan, J; Hanahoe, B; Cormican, M

2018-05-01

Lyme borreliosis is caused by Borrelia burgdorferi and is the most common tick-transmitted infection in temperate regions. Infection often presents with erythema migrans and/or other clinical features in early infection. Blood samples are submitted for testing for antibodies to Borrelia burgdorferi by enzyme immunoassay and positive samples are confirmed by a reference laboratory by IgG and IgM line immune assay. A retrospective extraction of all laboratory requests and results for Lyme borreliosis from 2011 to 2014 was performed. Patient addresses were mapped to local electoral area (LEA). The total number of requests was 5049 and 242 (5%) were positive over 5 years. The number of positive and tested samples were 40/748, 45/905, 41/947, 73/1126 and 43/1323 from 2011 to 2014. Even though the number of requests increased over the years, there was no significant increase in the number of positives. Incidences per 100,000 population for requests and positives were calculated at LEA level and showed considerable variation. The highest incidence was shown in one LEA (Connemara) with nearly 500 requests and 43 positives per 100,000 population per year. Increased awareness may explain the increase in requests. There is no indication of an increase in incidence. As many GPs treat suspected Lyme borreliosis empirically without testing and as antibody may be undetectable early in the course of illness, the true incidence of infection is likely to exceed the number of laboratory-confirmed cases.

Probability of detecting Porcine reproductive and respiratory syndrome virus infection using pen-based swine oral fluid specimens as a function of within-pen prevalence.

PubMed

Olsen, Chris; Wang, Chong; Christopher-Hennings, Jane; Doolittle, Kent; Harmon, Karen M; Abate, Sarah; Kittawornrat, Apisit; Lizano, Sergio; Main, Rodger; Nelson, Eric A; Otterson, Tracy; Panyasing, Yaowalak; Rademacher, Chris; Rauh, Rolf; Shah, Rohan; Zimmerman, Jeffrey

2013-05-01

Pen-based oral fluid sampling has proven to be an efficient method for surveillance of infectious diseases in swine populations. To better interpret diagnostic results, the performance of oral fluid assays (antibody- and nucleic acid-based) must be established for pen-based oral fluid samples. Therefore, the objective of the current study was to determine the probability of detecting Porcine reproductive and respiratory syndrome virus (PRRSV) infection in pen-based oral fluid samples from pens of known PRRSV prevalence. In 1 commercial swine barn, 25 pens were assigned to 1 of 5 levels of PRRSV prevalence (0%, 4%, 12%, 20%, or 36%) by placing a fixed number (0, 1, 3, 5, or 9) of PRRSV-positive pigs (14 days post PRRSV modified live virus vaccination) in each pen. Prior to placement of the vaccinated pigs, 1 oral fluid sample was collected from each pen. Thereafter, 5 oral fluid samples were collected from each pen, for a total of 150 samples. To confirm individual pig PRRSV status, serum samples from the PRRSV-negative pigs (n = 535) and the PRRSV vaccinated pigs (n = 90) were tested for PRRSV antibodies and PRRSV RNA. The 150 pen-based oral fluid samples were assayed for PRRSV antibody and PRRSV RNA at 6 laboratories. Among the 100 samples from pens containing ≥1 positive pig (≥4% prevalence) and tested at the 6 laboratories, the mean positivity was 62% for PRRSV RNA and 61% for PRRSV antibody. These results support the use of pen-based oral fluid sampling for PRRSV surveillance in commercial pig populations.

Occurrence and characterization of Shiga toxin-producing Escherichia coli in raw meat, raw milk, and street vended juices in Bangladesh.

PubMed

Islam, Mohammad A; Mondol, Abdus S; Azmi, Ishrat J; de Boer, Enne; Beumer, Rijkelt R; Zwietering, Marcel H; Heuvelink, Annet E; Talukder, Kaisar A

2010-11-01

The major objective of this study was to investigate the prevalence of Shiga toxin (Stx)-producing Escherichia coli (STEC) in different types of food samples and to compare their genetic relatedness with STEC strains previously isolated from animal sources in Bangladesh. We investigated a total of 213 food samples, including 90 raw meat samples collected from retail butcher shops, 20 raw milk samples from domestic cattle, and 103 fresh juice samples from street vendors in Dhaka city. We found that more than 68% (n = 62) of the raw meat samples were positive for the stx gene(s); 34% (n = 21) of buffalo meats and 66% (n = 41) of beef. Approximately 10% (n = 2) of the raw milk and 8% (n = 8) of the fresh juice samples were positive for stx. We isolated STEC O157 from seven meat samples (7.8%), of which two were from buffalo meats and five from beef; and no other STEC serotypes could be isolated. We could not isolate STEC from any of the stx-positive raw milk and juice samples. The STEC O157 isolates from raw meats were positive for the stx(2), eae, katP, etpD, and enterohemorrhagic E. coli hly virulence genes, and they belonged to three different phage types: 8 (14.3%), 31 (42.8%), and 32 (42.8%). Pulsed-field gel electrophoresis (PFGE) typing revealed six distinct patterns among seven isolates of STEC O157, suggesting a heterogeneous clonal diversity. Of the six PFGE patterns, one was identical and the other two were ≥90% related to PFGE patterns of STEC O157 strains previously isolated from animal feces, indicating that raw meats are readily contaminated with fecal materials. This study represents the first survey of STEC in the food chain in Bangladesh.

Detection of antileishmanial antibodies in blood sampled from blood bank donors in Istanbul.

PubMed

Ates, Sezen Canim; Bagirova, Malahat; Allahverdiyev, Adil M; Baydar, Serap Yesilkir; Koc, Rabia Cakir; Elcicek, Serhat; Abamor, Emrah Sefik; Oztel, Olga Nehir

2012-06-01

According to the WHO, only 5-20% of the total cases of leishmaniasis are symptomatic leishmaniasis; the other cases are identified as asymptomatic leishmaniasis. In recent studies, it has been demonstrated that donor blood plays an important role in the epidemiology of asymptomatic leishmaniasis. However, the number of the studies on this subject is still insufficient. Additionally, donor blood samples obtained from Istanbul, which is the biggest metropolitan area in Turkey, have not been investigated with regard to Leishmania. Moreover, there is no information about the sensitivity of noninvasive serological methods that are used in the detection of leishmaniasis donor blood samples. Accordingly, this study aimed to investigate the presence of antileishmanial antibodies in blood samples obtained from blood bank donors in Istanbul, by using different serologic methods, and to determine the most sensitive detection method. Blood samples were taken from 188 healthy blood bank donors to the Capa Turkish Red Crescent Blood Bank (Istanbul, Turkey), and the presence of antileishmanial antibodies was measured by indirect immunofluorescent antibody test (IFAT), ELISA, immunochromatographic dipstick rapid test, and western blot (WB). Antileishmanial antibodies were determined in 12 out of 188 samples by IFAT (6.4%), and six out of these 12 donors were found to be positive at diagnostic titer 1:128 (3.2%). One hundred and eighty eight samples were investigated by ELISA and one (0.5%) of them gave a positive result. None of 188 samples provided a positive result by immunochromatographic test. WB applied to the 12 seroreactive donors showed that three out of 12 donors were positive. In this study, the presence of antileishmanial antibodies in blood samples of blood bank donors from Istanbul has been demonstrated by using feasible and low-cost serological methods. Additionally, in comparison with other simple and low-cost detection methods, WB was used for confirmation. IFAT has a higher sensitivity and therefore may be preferred as a prescreening method in endemic or nonendemic areas.

Reciprocal contribution of clinical studies and the HP10 antigen ELISA for the diagnosis of extraparenchymal neurocysticercosis.

PubMed

Parkhouse, R Michael E; Carpio, Arturo; Campoverde, Alfredo; Sastre, Patricia; Rojas, Glenda; Cortez, María Milagros

2018-02-01

To evaluate diagnosis of active neurocysticercosis, paired cerebral spinal fluid (CSF) and serum samples from 24 neurocysticercosis (NCC) patients and 17 control neurological patients were assayed in the HP10 Taenia antigen (Ag) ELISA. The CSF samples were also tested with an HP10 Lateral Flow Assay (LFA). The HP10 Ag was detected by ELISA in the CSF of 5/5 patients with Definitive extraparenchymal NCC, and in 4/5 of the corresponding sera. In the Definitive parenchymal group, on the other hand, the HP10 Ag was absent in 2/3 CSF (with a very low value in the one positive sample) and all the corresponding serum samples. Samples of CSF from 4/7 patients in the Probable parenchymal group, were also significantly HP10 Ag positive, suggesting the presence of extraparenchymal cysts not identified by the imaging studies. With the possible exception of one patient, the corresponding serum samples of the Probable parenchymal NCC group, were all HP10 Ag negative. Samples of CSF from 9 NCC patients diagnosed with Mixed parenchymal and extraparenchymal NCC were all significantly HP10 Ag positive, confirming the presence of extraparenchymal cysts, with only 7/9 of the corresponding serum samples being HP10 positive. Thus detection of the HP10 Ag indicates extraparenchymal and not parenchymal cyst localization and is more sensitive with CSF than serum. Three neurological patients clinically diagnosed as subarachnoid cyst, hydrocephalus and tuberculoma, respectively, were clearly positive for HP10 Ag. Of these, two were confirmed as NCC by subsequent imaging; the third died prior to further examination. Thus, a total of 8 patients had their clinical diagnosis questioned. Finally, there was good agreement between the HP10 Ag ELISA and LFA with CSF samples giving an optical density ≥0.4 in the ELISA assay. In conclusion, the HP10 Ag assay should provide a valuable and reciprocal tool in the clinical diagnosis and follow up of extraparenchymal NCC. Copyright © 2017 Elsevier B.V. All rights reserved.

Detection of Tick-Borne Pathogens in Lambs Undergoing Prophylactic Treatment Against Ticks on Two Swedish Farms.

PubMed

Grandi, Giulio; Aspán, Anna; Pihl, Jenny; Gustafsson, Katarina; Engström, Fredrik; Jinnerot, Tomas; Söderlund, Robert; Chirico, Jan

2018-01-01

Tick-borne pathogens (TBPs), especially Anaplasma phagocytophilum , cause disease in grazing livestock. Tick prophylaxis is, therefore, a routine practice in sheep flocks in Sweden, especially in central, southern, and coastal areas of the country where ixodid ticks ( Ixodes ricinus and Haemaphysalis punctata ) are present. In the present study, the status of infection by A. phagocytophilum and other TBPs in lambs treated with tick prophylaxis has been assessed serologically and with polymerase chain reaction (PCR). Blood samples ( n  = 78) from lambs ( n  = 20) subjected to regular tick prophylactic treatment (flumethrin, Bayticol ® ) at two sites in different regions in Sweden (Östergötland, Gotland) were collected on four occasions from May until July 2013. The severity of clinical signs in Anaplasma- infected animals is known to differ between these two regions. In total, 20% of blood samples were PCR-positive for A. phagocytophilum . Serological analyses showed that 33% of all collected samples were positive for A. phagocytophilum , while 2.5% were positive for Borrelia burgdorferi s.l. and 13% for tick-borne encephalitis virus (TBEV). Percentages of lambs positive were 75 and 45% for A. phagocytophilum antibodies and DNA, respectively, while 10 and 45% were serologically positive for B. burgdorferi s.l. and TBEV, respectively. Sequencing of partial 16S rRNA genes from Anaplasma PCR positive samples revealed presence of A. phagocytophilum in all animals in Östergötland, while sequences consistent with A. phagocytophilum as well as A. capra and A. bovis were found on the island of Gotland. This is the first report of the occurrence of the latter two species in Sweden.