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Sample records for enteric viruses coliphages

  1. Water quality indicators: bacteria, coliphages, enteric viruses.

    PubMed

    Lin, Johnson; Ganesh, Atheesha

    2013-12-01

    Water quality through the presence of pathogenic enteric microorganisms may affect human health. Coliform bacteria, Escherichia coli and coliphages are normally used as indicators of water quality. However, the presence of above-mentioned indicators do not always suggest the presence of human enteric viruses. It is important to study human enteric viruses in water. Human enteric viruses can tolerate fluctuating environmental conditions and survive in the environment for long periods of time becoming causal agents of diarrhoeal diseases. Therefore, the potential of human pathogenic viruses as significant indicators of water quality is emerging. Human Adenoviruses and other viruses have been proposed as suitable indices for the effective identification of such organisms of human origin contaminating water systems. This article reports on the recent developments in the management of water quality specifically focusing on human enteric viruses as indicators.

  2. Evaluation of FRNA coliphages as indicators of human enteric viruses in a tropical urban freshwater catchment.

    PubMed

    Vergara, G G R V; Goh, S G; Rezaeinejad, S; Chang, S Y; Sobsey, M D; Gin, K Y H

    2015-08-01

    This study aimed to evaluate the relationship between FRNA coliphages (FRNA GI to GIV) and human enteric viruses (human adenoviruses, HAdV, astroviruses, AstV, noroviruses, NoV, and rotaviruses, RoV) in a tropical urban freshwater catchment. Positive associations between human-specific coliphages and human viral pathogens substantiate their use as viral indicators and in microbial source tracking. Reverse transcription qPCR was used to measure the concentrations of viruses and FRNA coliphages in concentrated water samples. Environmental water samples were also analyzed for male-specific (F+) and somatic (Som) coliphages using plaque assay. The most abundant enteric virus was NoV (55%) followed by HAdV (33%), RoV (33%), and AstV (23%), while the most abundant FRNA genogroup was GI (85%) followed by GII (48%), GIV (8%) and GIII (7%). Concentrations of human-specific coliphages FRNA GII were positively correlated with NoV, HAdV, RoV, AstV, F+ and Som (τ = 0.5 to 0.3, P < 0.05) while concentrations of animal-specific coliphages FRNA GI were negatively correlated with HAdV and RoV (τ = -0.2, P < 0.05). This study demonstrates statistical relationships between human-specific coliphages and a suite of human enteric viruses in the environment.

  3. Assessment of F-RNA coliphage as a potential indicator of enteric virus contamination of hog carcasses.

    PubMed

    Jones, T H; Johns, M W

    2012-08-01

    Hepatitis E virus (HEV) is common in pigs, and some swine HEV strains are closely related to human strains. The zoonotic transmission of HEV is now well established. HEV can be detected by molecular techniques, but the significance of the presence of viral nucleic acid is questionable when foods are subjected to virus inactivation treatments. F-RNA coliphages are attractive candidates as indicators for enteric viruses because they are similar in size and survival characteristics and can be rapidly cultured. Information on the contamination of hog carcasses with enteric or hepatic viruses during slaughter is lacking. The objective of this study was to compare the incidence and levels of contamination of hog carcasses with F-RNA coliphages, HEV, total aerobic bacteria, coliforms, and Escherichia coli at different stages of the dressing process. Hog carcasses entering the commercial slaughter facility are heavily contaminated with F-RNA coliphages and HEV. Subsequent processes such as scalding, singing, and pasteurization can reduce the incidence and levels of F-RNA coliphages and HEV substantially to almost undetectable levels. Large discrepancies between the amount of viral nucleic acid and infectious F-RNA coliphage particles, both at high levels and low levels of contamination, were observed. The prevalence and levels of viable F-RNA coliphages were lower than those of total aerobic bacteria, coliforms, and E. coli in the anal area and on random sites before pasteurization. At a research abattoir, there was no overall mean reduction of viable F-RNA coliphages recovered from random sites before pasteurization and after washing, whereas overall mean reductions of 1.2, 2.6, and 2.9 log CFU for total aerobic bacteria, coliforms, and E. coli, respectively, were obtained. These findings suggest that bacteria such as coliforms and E. coli may not be suitable as indicators for enteric viruses in a meat processing environment.

  4. Comparative study of enteric viruses, coliphages and indicator bacteria for evaluating water quality in a tropical high-altitude system

    PubMed Central

    2009-01-01

    Background Bacteria used as indicators for pathogenic microorganisms in water are not considered adequate as enteric virus indicators. Surface water from a tropical high-altitude system located in Mexico City that receives rainwater, treated and non-treated wastewater used for irrigation, and groundwater used for drinking, was studied. Methods The presence of enterovirus, rotavirus, astrovirus, coliphage, coliform bacteria, and enterococci was determined during annual cycles in 2001 and 2002. Enteric viruses in concentrated water samples were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Coliphages were detected using the double agar layer method. Bacteria analyses of the water samples were carried out by membrane filtration. Results The presence of viruses and bacteria in the water used for irrigation showed no relationship between current bacterial indicator detection and viral presence. Coliphages showed strong association with indicator bacteria and enterovirus, but weak association with other enteric viruses. Enterovirus and rotavirus showed significant seasonal differences in water used for irrigation, although this was not clear for astrovirus. Conclusion Coliphages proved to be adequate faecal pollution indicators for the irrigation water studied. Viral presence in this tropical high-altitude system showed a similar trend to data previously reported for temperate zones. PMID:19860917

  5. Evaluation of an anion exchange resin-based method for concentration of F-RNA coliphages (enteric virus indicators) from water samples.

    PubMed

    Pérez-Méndez, A; Chandler, J C; Bisha, B; Goodridge, L D

    2014-08-01

    Enteric viral contaminants in water represent a public health concern, thus methods for detecting these viruses or their indicator microorganisms are needed. Because enteric viruses and their viral indicators are often found at low concentrations in water, their detection requires upfront concentration methods. In this study, a strong basic anion exchange resin was evaluated as an adsorbent material for the concentration of F-RNA coliphages (MS2, Qβ, GA, and HB-P22). These coliphages are recognized as enteric virus surrogates and fecal indicator organisms. Following adsorption of the coliphages from 50ml water samples, direct RNA isolation and real time RT-PCR detection were performed. In water samples containing 10(5)pfu/ml of the F-RNA coliphages, the anion exchange resin (IRA-900) adsorbed over 96.7% of the coliphages present, improving real time RT-PCR detection by 5-7 cycles compared to direct testing. F-RNA coliphage RNA recovery using the integrated method ranged from 12.6% to 77.1%. Resin-based concentration of samples with low levels of the F-RNA coliphages allowed for 10(0)pfu/ml (MS2 and Qβ) and 10(-1)pfu/ml (GA and HB-P22) to be detected. The resin-based method offers considerable advantages in cost, speed, simplicity and field adaptability.

  6. Removal of indigenous coliphages and enteric viruses during riverbank filtration from highly polluted river water in Delhi (India).

    PubMed

    Sprenger, C; Lorenzen, G; Grunert, A; Ronghang, M; Dizer, H; Selinka, H-C; Girones, R; Lopez-Pila, J M; Mittal, A K; Szewzyk, R

    2014-06-01

    Emerging countries frequently afflicted by waterborne diseases require safe and cost-efficient production of drinking water, a task that is becoming more challenging as many rivers carry a high degree of pollution. A study was conducted on the banks of the Yamuna River, Delhi, India, to ascertain if riverbank filtration (RBF) can significantly improve the quality of the highly polluted surface water in terms of virus removal (coliphages, enteric viruses). Human adenoviruses and noroviruses, both present in the Yamuna River in the range of 10(5) genomes/100 mL, were undetectable after 50 m infiltration and approximately 119 days of underground passage. Indigenous somatic coliphages, used as surrogates of human pathogenic viruses, underwent approximately 5 log10 removal after only 3.8 m of RBF. The initial removal after 1 m was 3.3 log10, and the removal between 1 and 2.4 m and between 2.4 and 3.8 m was 0.7 log10 each. RBF is therefore an excellent candidate to improve the water situation in emerging countries with respect to virus removal.

  7. Preliminary Source Tracking of Male-Specific (F(+)) RNA Coliphage on Lettuce as a Surrogate of Enteric Viruses Using Reverse Transcription-PCR.

    PubMed

    Yazdi, Mojgan; Yavarmanesh, Masoud; Bahreini, Masumeh; Mohebbi, Mohebbat

    2017-03-01

    The aim of this research was to preliminary track fecal source male-specific F(+)RNA coliphages including human and animals in lettuce. At first, two published virus extraction procedures of ultracentrifugation and PEG precipitation were compared using DAL assay for determining the recovery efficiency in lettuce spiked artificially with three concentrations (10(2), 10(4), 10(6) pfu/100 ml) of MS2 coliphage. The results showed that PEG precipitation had the highest recovery in which the recovery efficiency at the spiked level of 10(6) pfu/100 ml was 16.63 %. Aqueous phase obtained from the final step of PEG method was applied for enumeration of coliphage and viral RNA extraction in naturally contaminated lettuce samples (N = 30) collected from two sources (market and farm). The samples were then analyzed based on (I, II, III, and IV primer sets) using RT-PCR method. Coliphages were detected in 9 (60 %) and 12 (80 %) out of 15 market and farm samples, respectively, using DAL assay, whereas male-specific F(+)RNA coliphages were detected using the RT-PCR method in 9 (60 %) and 13 (86.6 %) out of 15 samples of market and farm, respectively. Based on the results, only genotype I of male-specific F(+)RNA coliphages was detected in lettuce samples and no sample tested was positive for other genotypes (II, III, and IV).

  8. Application of F⁺RNA Coliphages as Source Tracking Enteric Viruses on Parsley and Leek Using RT-PCR.

    PubMed

    Shahrampour, Dina; Yavarmanesh, Masoud; Najafi, Mohammad Bagher Habibi; Mohebbi, Mohebbat

    2015-12-01

    The objective of this study was to identify sources of fecal contamination in leek and parsley, by using four different F(+)RNA coliphage genogroups (IV, I indicate animal fecal contamination and II, III indicate human fecal contamination). Three different concentrations (10(2), 10(4), 10(6) pfu/ml) of MS2 coliphage were inoculated on the surface of parsley and leek samples for detection of phage recovery efficiency among two methods of elution concentration (PEG-precipitation and Ultracentrifugation) by performing double agar layer (DAL) assay in three replications. Highest recovery of MS2 was observed in PEG method and in 10(6) inoculation concentration. Accordingly, the PEG method was used for washing and isolation of potentially contaminated phages of 30 collected samples (15 samples from the market and 15 samples from the farm). The final solutions of PEG method were tested for the enumeration of plaques by DAL assay. Total RNA was then extracted from recovered phages, and RT-PCR was performed by using four primer sets I, II, III, and IV. Incidence of F(+)RNA coliphages was observed in 12/15 (80 %) and 10/15 (66/6 %) of samples were obtained from farm and market, respectively, using both DAL and RT-PCR test methods. Different genotypes (I, II, and IV) of F(+)RNA coliphages were found in farm samples, while only genotype I was detected in market samples by using the primer sets. Due to the higher frequency of genotype I and IV, the absence of genotype III, and also the low frequency of genotype II, it is concluded that the contamination of vegetable (parsley and leek) in Neyshabour, Iran is most likely originated from animal sources.

  9. Inactivation of a model coliphage virus in water by iodine

    NASA Technical Reports Server (NTRS)

    Brion, Gail M.; Silverstein, Joann

    1992-01-01

    Until now, NASA's space water reuse research program has not considered the transport of water-borne infectious enteric viruses; however, viral diseases probably are a signifficant concern in long-duration space missions. To simplify monitoring and prediction of pathogen distribution, model indicator strains historically have been used. In this research, the male specific RNA coliphage MS-2 is used as a model of enteric viruses due to their similar size and biochemical composition. Inactivation of some water-borne enteric viruses by iodine has previously been characterized. In this paper, iodine inactivation of the model coliphage MS-2 in buffered water is compared with earlier bench-scale disinfection survival data and with survival in iodinated simulated shower water used in a test water recycling system.

  10. 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.

  11. F-coliphages, porcine adenovirus and porcine teschovirus as potential indicator viruses of fecal contamination for pork carcass processing.

    PubMed

    Jones, Tineke H; Muehlhauser, Victoria

    2017-01-16

    There are concerns about the zoonotic transmission of viruses through undercooked pork products. There is a lack of information on suitable indicator viruses for fecal contamination with pathogenic enteric viruses in the meat processing chain. The study compared the incidence and levels of contamination of hog carcasses with F-coliphages, porcine teschovirus (PTV), and porcine adenovirus (PAdV) at different stages of the dressing process to assess their potential as indicator viruses of fecal contamination. One hundred swab samples (200cm(2)) were collected from random sites on hog carcasses at 4 different stages of the dressing process and from retail pork over the span of a year from 2 pork processing plants (500/plant). Viable F-coliphages, PAdV DNA and PTV RNA were each detected on ≥99% of the incoming carcasses at both plants and were traceable through the pork processing chain. Significant correlations were observed between viable F-coliphages and PAdV DNA and between F-coliphages and PTV RNA but not between PAdV DNA and PTV RNA at the various stages of pork processing. Detection of viable F-coliphages was more sensitive than genomic copies of PAdV and PTV at low levels of contamination, making F-coliphages a preferred indicator in the pork slaughter process as it also provides an indication of infectivity. For plant A, F-RNA coliphages were detected in 25%, 63%, and 21% of carcass swabs after pasteurization, evisceration, and retail pork products, respectively. For plant B, F-coliphages were detected in 33%, 25%, and 13% of carcass swabs after skinning, evisceration, and retail pork samples, respectively. Viable F-RNA coliphages were genotyped. Viable F-RNA GII and GIII were generally not detected at the earlier stages of the slaughter process but they were detected on 13% of carcasses after evisceration and 2% of retail pork samples at plant A, which raises concerns of potential food handler contamination during pork processing. Consumers could be at risk

  12. Animal viruses, coliphages, and bacteria in aerosols and wastewater at a spray irrigation site.

    PubMed Central

    Brenner, K P; Scarpino, P V; Clark, C S

    1988-01-01

    Aerosol samples collected at the Muskegon County Wastewater Management System Number 1 spray irrigation site in Michigan by using the Army prototype XM2 Biological Sampler/Collector were examined for the presence of animal viruses, coliphages, and bacteria. Air samples, collected in Earle lactalbumen hydrolysate, and wastewater samples were filtered through a 0.45- and 1.2-micron membrane filter sandwich, pretreated with 10% beef extract (pH 7.0), and assayed for animal viruses by the plaque method on Buffalo green monkey kidney cells. Untreated air and wastewater samples were assayed for coliphages by the soft agar overlay method with three Escherichia coli hosts (ATCC 13706, 15597, and 11303) and for bacteria by the heterotrophic plate count method. Filtered air samples were assayed for coliphages by the most-probable-number method with the same three hosts. Although no animal viruses were detected in the aerosol samples, coliphages and bacteria were recovered. E. coli ATCC 13706 coliphage were recovered more often and in greater numbers than either of the other two types of coliphages. Concentrations of animal viruses, coliphages, and bacteria detected in the raw influent decreased as the wastewater was aerated and stored in the lagoons. No animal viruses were detected in the wastewater at the pump station just before distribution to the spray irrigation rigs. The most-probable-number method was more sensitive and consistent than the overlay procedure in detecting low levels of coliphages in air samples. PMID:3128164

  13. Animal viruses, coliphages, and bacteria in aerosols and wastewater at a spray irrigation site.

    PubMed

    Brenner, K P; Scarpino, P V; Clark, C S

    1988-02-01

    Aerosol samples collected at the Muskegon County Wastewater Management System Number 1 spray irrigation site in Michigan by using the Army prototype XM2 Biological Sampler/Collector were examined for the presence of animal viruses, coliphages, and bacteria. Air samples, collected in Earle lactalbumen hydrolysate, and wastewater samples were filtered through a 0.45- and 1.2-micron membrane filter sandwich, pretreated with 10% beef extract (pH 7.0), and assayed for animal viruses by the plaque method on Buffalo green monkey kidney cells. Untreated air and wastewater samples were assayed for coliphages by the soft agar overlay method with three Escherichia coli hosts (ATCC 13706, 15597, and 11303) and for bacteria by the heterotrophic plate count method. Filtered air samples were assayed for coliphages by the most-probable-number method with the same three hosts. Although no animal viruses were detected in the aerosol samples, coliphages and bacteria were recovered. E. coli ATCC 13706 coliphage were recovered more often and in greater numbers than either of the other two types of coliphages. Concentrations of animal viruses, coliphages, and bacteria detected in the raw influent decreased as the wastewater was aerated and stored in the lagoons. No animal viruses were detected in the wastewater at the pump station just before distribution to the spray irrigation rigs. The most-probable-number method was more sensitive and consistent than the overlay procedure in detecting low levels of coliphages in air samples.

  14. Clostridium perfringens and somatic coliphages as indicators of the efficiency of drinking water treatment for viruses and protozoan cysts.

    PubMed

    Payment, P; Franco, E

    1993-08-01

    To find the most suitable indicator of viral and parasitic contamination of drinking water, large-volume samples were collected and analyzed for the presence of pathogens (cultivable human enteric viruses, Giardia lamblia cysts, and Cryptosporidium oocysts) and potential indicators (somatic and male-specific coliphages, Clostridium perfringens). The samples were obtained from three water treatment plants by using conventional or better treatments (ozonation, biological filtration). All samples of river water contained the microorganisms sought, and only C. perfringens counts were correlated with human enteric viruses, cysts, or oocysts. For settled and filtered water samples, all indicators were statistically correlated with human enteric viruses but not with cysts or oocysts. By using multiple regression, the somatic coliphage counts were the only explanatory variable for the human enteric virus counts in settled water, while in filtered water samples it was C. perfringens counts. Finished water samples of 1,000 liters each were free of all microorganisms, except for a single sample that contained low levels of cysts and oocysts of undetermined viability. Three of nine finished water samples of 20,000 liters each revealed residual levels of somatic coliphages at 0.03, 0.10, and 0.26 per 100 liters. Measured virus removal was more than 4 to 5 log10, and cyst removal was more than 4 log10. Coliphage and C. perfringens counts suggested that the total removal and inactivation was more than 7 log10 viable microorganisms. C. perfringens counts appear to be the most suitable indicator for the inactivation and removal of viruses in drinking water treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

  15. Clostridium perfringens and somatic coliphages as indicators of the efficiency of drinking water treatment for viruses and protozoan cysts.

    PubMed Central

    Payment, P; Franco, E

    1993-01-01

    To find the most suitable indicator of viral and parasitic contamination of drinking water, large-volume samples were collected and analyzed for the presence of pathogens (cultivable human enteric viruses, Giardia lamblia cysts, and Cryptosporidium oocysts) and potential indicators (somatic and male-specific coliphages, Clostridium perfringens). The samples were obtained from three water treatment plants by using conventional or better treatments (ozonation, biological filtration). All samples of river water contained the microorganisms sought, and only C. perfringens counts were correlated with human enteric viruses, cysts, or oocysts. For settled and filtered water samples, all indicators were statistically correlated with human enteric viruses but not with cysts or oocysts. By using multiple regression, the somatic coliphage counts were the only explanatory variable for the human enteric virus counts in settled water, while in filtered water samples it was C. perfringens counts. Finished water samples of 1,000 liters each were free of all microorganisms, except for a single sample that contained low levels of cysts and oocysts of undetermined viability. Three of nine finished water samples of 20,000 liters each revealed residual levels of somatic coliphages at 0.03, 0.10, and 0.26 per 100 liters. Measured virus removal was more than 4 to 5 log10, and cyst removal was more than 4 log10. Coliphage and C. perfringens counts suggested that the total removal and inactivation was more than 7 log10 viable microorganisms. C. perfringens counts appear to be the most suitable indicator for the inactivation and removal of viruses in drinking water treatment.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8368831

  16. Enteric viruses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Characteristic clinical signs associated with viral enteritis in young poultry include diarrhea, anorexia, litter eating, ruffled feathers, and poor growth. Intestines may have lesions; intestines are typically dilated and are filled with fluid and gaseous contents. The sequela to clinical disease...

  17. A fate model of pathogenic viruses in a composting toilet based on coliphage inactivation.

    PubMed

    Kazama, Shinobu; Tameike, Narue; Nakagawa, Naoko; Otaki, Masahiro

    2011-01-01

    A composting toilet using sawdust as a matrix has the potential to trap pathogens that might occasionally be contained in human feces. Therefore, care should be taken when handling the sawdust. It should also be noted that pathogenic viruses tend to have stronger tolerance than pathogenic bacteria. The fates of several species of coliphages, T4, lambda, Qbeta and MS2, in sawdust were investigated as a viral model. The fates of coliphages were significantly different among them, and they changed in response to temperature and the water content of the sawdust. As the results, T4 coliphage had the strongest tolerance and Qbeta had the weakest one in sawdust. It was estimated the days required to decrease virus to a safe level based on a risk assessment. According to the rates of Qbeta and T4, 15 days and 167 days were required respectively for a safe level of infection risk based on actually operated composting toilet condition. Thus, it was significantly different depending on the species and sawdust conditions.

  18. Incidence of Somatic and F+ Coliphage at Three Great Lake Beaches

    EPA Science Inventory

    There is a growing interest for the potential use of coliphage as an alternative indicator to assess fecal pollution in recreational waters. Coliphage are a group of viruses that infect E. coli and are commonly used as models to infer the likely presence of human enteric viral pa...

  19. Bacteriophages as indicators of fecal pollution and enteric virus removal.

    PubMed

    McMinn, Brian R; Ashbolt, Nicholas J; Korajkic, Asja

    2017-03-17

    Bacteriophages are an attractive alternative to fecal indicator bacteria (FIB), particularly as surrogates of enteric virus fate and transport, due to their closer morphological and biological properties. Based on a review of published data, we summarize densities of coliphages (F+ and somatic), Bacteroides spp. and enterococci bacteriophages (phages) in individual human waste, raw wastewater, ambient fresh and marine waters and removal through wastewater treatment processes utilizing traditional treatments. We also provide comparisons with FIB and enteric viruses whenever possible. Lastly, we examine fate and transport characteristics in the aquatic environment and provide an overview of the environmental factors affecting their survival. In summary, concentrations of bacteriophages in various sources were consistently lower than FIB, but more reflective of infectious enteric virus levels. Overall, our investigation indicates that bacteriophages may be adequate viral surrogates, especially in built systems, such as wastewater treatment plants. This article is protected by copyright. All rights reserved.

  20. In situ inactivation of animal viruses and a coliphage in nonaerated liquid and semiliquid animal wastes.

    PubMed

    Pesaro, F; Sorg, I; Metzler, A

    1995-01-01

    The persistence of five animal viruses, representing picorna-, rota-, parvo-, adeno-, and herpesviruses, and the coliphage f2 was determined in the field by exposing the viruses to different animal wastes and by adopting an established filter sandwich technique. This technique allows us to copy the natural state of viruses in the environment, where adsorption onto or incorporation into suspended solids may prolong virus survival. Using filter sandwiches either equipped with porous (15 nm in diameter) or poreless polycarbonate (PC) membranes, it was possible to differentiate between overall virus inactivation and the effect of virucidal agents that act through poreless PC membranes. Depending on ambient temperature, pH, and type of animal waste, values for time, in days, required for a 90% reduction of virus titer varied widely, ranging from less than 1 week for herpesvirus to more than 6 months for rotavirus. Virus inactivation progressed substantially faster in liquid cattle manure, a mixture of urine and water (pH > 8.0), than in semiliquid wastes that consisted of mixtures of feces, urine, water, and bedding materials (pH < 8.0). Hitherto unidentified virucidal agents that permeate poreless PC membranes contributed substantially to the overall inactivation. On the other hand, substances that protect rotavirus and possibly other viruses from inactivation may be present in animal wastes. Together, the study showed that viruses contained in manure may persist for prolonged periods of time if stored under nonaerated conditions. At times of land application, this may lead to environmental contamination with pathogens.

  1. Waterborne Viruses and F-Specific Coliphages in Mixed-Use Watersheds: Microbial Associations, Host Specificities, and Affinities with Environmental/Land Use Factors.

    PubMed

    Jones, Tineke H; Brassard, Julie; Topp, Edward; Wilkes, Graham; Lapen, David R

    2017-02-01

    From the years 2008 to 2014, a total of 1,155 water samples were collected (spring to fall) from 24 surface water sampling sites located in a mixed-used but predominantly agricultural (i.e., dairy livestock production) river basin in eastern Ontario, Canada. Water was analyzed for viable F-specific DNA (F-DNA) and F-specific RNA (F-RNA) (genogroup I [GI] to GIV) coliphage and a suite of molecularly detected viruses (norovirus [GI to GIV], torque teno virus [TTV], rotavirus, kobuvirus, adenovirus, astrovirus, hepatitis A, and hepatitis E). F-DNA and F-RNA coliphage were detected in 33 and 28% of the samples at maximum concentrations of 2,000 and 16,300 PFU · 100 ml(-1), respectively. Animal TTV, human TTV, kobuvirus, astrovirus, and norovirus GIII were the most prevalent viruses, found in 23, 20, 13, 12, and 11% of samples, respectively. Viable F-DNA coliphage was found to be a modest positive indicator of molecularly detected TTV. F-RNA coliphage, unlike F-DNA coliphage, was a modest positive predictor of norovirus and rotavirus. There were, however, a number of significant negative associations among F-specific coliphage and viruses. F-DNA coliphage densities of >142 PFU · 100 ml(-1) delineated conditions when ∼95% of water samples contained some type of virus. Kobuvirus was the virus most strongly related to detection of any other virus. Land use had some associations with virus/F-specific coliphage detection, but season and surface water flow were the variables that were most important for broadly delineating detection. Higher relative levels of detection of human viruses and human F-RNA coliphage were associated with higher relative degrees of upstream human land development in a catchment.

  2. Removal of human enteric viruses and indicator microorganisms from domestic wastewater by aerated lagoons.

    PubMed

    Locas, Annie; Martinez, Veronica; Payment, Pierre

    2010-02-01

    Aerated lagoons offer a low-cost and simple approach to treating domestic wastewater in small municipalities. The objective of the current study was to evaluate, for each cell in the lagoons, the removal of indicator microorganisms and human enteric viruses under warm (summer) and cold (early spring) conditions. The two sites are located in southwest Quebec, Canada. Samples were assayed for thermotolerant coliforms, enterococci, Clostridium perfringens, somatic and male-specific coliphages, and culturable human enteric viruses (HEV). The results show higher removal under warm ambient conditions for all microorganisms. Thermotolerant coliforms and enterococci were removed to a greater extent than C. perfringens and HEV. HEV removal was only observed in warm ambient conditions. The removal of coliphages was different from the observed removal of HEV. The use of coliphages as surrogates for HEV has been proposed, but this does not seem appropriate for aerated lagoons, as the removal of coliphages overestimates the removal of HEV. Given the low observed removal of HEV during this study, the effluents remain a significant source of pathogens that can affect drinking water treatment plants drawing their raw water from receiving streams. Ultraviolet disinfection of treated wastewater effluent is a possible solution.

  3. Enteric viruses of poultry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Despite the economic importance of the poultry gut, very little is known about the complex gut microbial community. Enteric disease syndromes such as Runting-Stunting Syndrome (RSS) in broiler chickens and Poult Enteritis Complex (PEC) in young turkeys are difficult to characterize and reproduce in ...

  4. Pathogenic Enteric Viruses and Microbial Indicators during Secondary Treatment of Municipal Wastewater.

    PubMed

    Montazeri, Naim; Goettert, Dorothee; Achberger, Eric C; Johnson, Crystal N; Prinyawiwatkul, Witoon; Janes, Marlene E

    2015-09-01

    Pathogenic enteric viruses are responsible for a wide range of infections in humans, with diverse symptoms. Raw and partially treated wastewaters are major sources of environmental contamination with enteric viruses. We monitored a municipal secondary wastewater treatment plant (New Orleans, LA) on a monthly basis for norovirus (NoV) GI and GII and enterovirus serotypes using multiplex reverse transcription-quantitative PCR (RT-qPCR) and microbial indicators of fecal contamination using standard plating methods. Densities of indicator bacteria (enterococci, fecal coliforms, and Escherichia coli) did not show monthly or seasonal patterns. Norovirus GII was more abundant than GI and, along with enterovirus serotypes, increased in influent during fall and spring. The highest NoV GI density in influent was in the fall, reaching an average of 4.0 log10 genomic copies/100 ml. Norovirus GI removal (0.95 log10) was lower than that for GII, enterovirus serotypes, and male-specific coliphages (1.48 log10) or for indicator bacteria (4.36 log10), suggesting higher resistance of viruses to treatment. Male-specific coliphages correlated with NoV GII densities in influent and effluent (r = 0.48 and 0.76, respectively) and monthly removal, indicating that male-specific coliphages can be more reliable than indicator bacteria to monitor norovirus GII load and microbial removal. Dominant norovirus genotypes were classified into three GI genotypes (GI.1, GI.3, and GI.4) and four GII genotypes (GII.3, GII.4, GII.13, and GII.21), dominated by GI.1 and GII.4 strains. Some of the seasonal and temporal patterns we observed in the pathogenic enteric viruses were different from those of epidemiological observations.

  5. Assessment of coliphage surrogates for testing drinking water treatment devices.

    PubMed

    Gerba, Charles P; Abd-Elmaksoud, Sherif; Newick, Huikheng; El-Esnawy, Nagwa A; Barakat, Ahmed; Ghanem, Hossam

    2015-03-01

    Test protocols have been developed by the United States Environmental Protection Agency (USEPA) and the World Health Organization (WHO) to test water treatment devices/systems that are used at the individual and home levels to ensure the removal of waterborne viruses. The goal of this study was to assess if coliphage surrogates could be used in this testing in place of the currently required use of animal or human enteric viruses. Five different coliphages (MS-2, PRD1, ΦX-174, Qβ, and fr) were compared to the removal of poliovirus type 1 (LSc-2ab) by eight different water treatment devices/systems using a general case and a challenge case (high organic load, dissolved solids, and turbidity) test water as defined by the USEPA. The performance of the units was rated as a pass/fail based on a 4 log removal/inactivation of the viruses. In all cases, a failure or a pass of the units/system for poliovirus also corresponded to a pass/fail by all of the coliphages. In summary, in using pass/fail criteria as recommended under USEPA guidelines for testing water treatment device/systems, the use of coliphages should be considered as an alternative to reduce cost and time of testing such devices/systems.

  6. Removal of bacterial fecal indicators, coliphages and enteric adenoviruses from waters with high fecal pollution by slow sand filtration.

    PubMed

    Bauer, Rosalie; Dizer, Halim; Graeber, Ingeborg; Rosenwinkel, Karl-Heinz; López-Pila, Juan M

    2011-01-01

    The aim of the present study was to estimate the performance of slow sand filtration (SSF) facilities, including the time needed for reaching stabilization (maturation), operated with surface water bearing high fecal contamination, representing realistic conditions of rivers in many emerging countries. Surface water spiked with wastewater was infiltrated at different pore water velocities (PWV) and samples were collected at different migration distances. The samples were analyzed for phages and to a lesser extent for fecal bacteria and enteric adenoviruses. At the PWV of 50 cm/d, at which somatic phages showed highest removal, their mean log(10) removal after 90 cm migration was 3.2. No substantial differences of removal rates were observed at PWVs between 100 and 900 cm/d (2.3 log(10) mean removal). The log(10) mean removal of somatic phages was less than the observed for fecal bacteria and tended more towards that of enteric adenoviruses This makes somatic phages a potentially better process indicator than Escherichia coli for the removal of viruses in SSF. We conclude that SSF, and by inference in larger scale river bank filtration (RBF), is an excellent option as a component in multi-barrier systems for drinking water treatment also in areas where the sources of raw water are considerably fecally polluted, as often found in many emerging countries.

  7. Human enteric viruses--potential indicators for enhanced monitoring of recreational water quality.

    PubMed

    Updyke, Erin Allmann; Wang, Zi; Sun, Si; Connell, Christina; Kirs, Marek; Wong, Mayee; Lu, Yuanan

    2015-10-01

    Recreational waters contaminated with human fecal pollution are a public health concern, and ensuring the safety of recreational waters for public use is a priority of both the Environmental Protection Agency (EPA) and the Centers for Disease Control and Prevention (CDC). Current recreational water standards rely on fecal indicator bacteria (FIB) levels as indicators of human disease risk. However present evidence indicates that levels of FIB do not always correspond to the presence of other potentially harmful organisms, such as viruses. Thus, enteric viruses are currently tested as water quality indicators, but have yet to be successfully implemented in routine monitoring of water quality. This study utilized enteric viruses as possible alternative indicators of water quality to examine 18 different fresh and offshore recreational waters on O'ahu, Hawai'i, by using newly established laboratory techniques including highly optimized PCR, real time PCR, and viral infectivity assays. All sample sites were detected positive for human enteric viruses by PCR including enterovirus, norovirus genogroups I and II, and male specific FRNA coliphage. A six time-point seasonal study of enteric virus presence indicated significant variation in virus detection between the rainy and dry seasons. Quantitative PCR detected the presence of norovirus genogroup II at levels at which disease risk may occur, and there was no correlation found between enteric virus presence and FIB counts. Under the present laboratory conditions, no infectious viruses were detected from the samples PCR-positive for enteric viruses. These data emphasize both the need for additional indicators for improved monitoring of water quality, and the feasibility of using enteric viruses as these indicators.

  8. Effective reduction of enteric bacteria and viruses during the anaerobic digestion of biomass and wastes

    SciTech Connect

    Fannin, K.F.; Hsu, P.H.; Mensinger, J.; Cahill, C.

    1984-01-01

    Natural resource depletion increases the amount of waste requiring efficient and affordable disposal alternatives. Through effective management, many of these so-called wastes can be utilized as important energy and agricultural resources. One such management approach involves the utilization of emergent aquatic plant species, such as water hyacinth, to remove nutrients from the wastewater during growth. This process produces an energy-containing biomass that can then be anaerobically digested either separately or with other waste components to produce energy-containing methane and an effluent residue containing significant quantities of protein and nutrients. This residue can be utilized as an effective fertilizer, soil conditioner, or animal feed supplement provided it is rendered reasonably safe from such contaminants as enteric microorganisms. This study was conducted to identify the digester operating parameters that affect the survival of enteric bacteria and viruses during the anaerobic digestion of blends of water hyacinth and primary sewage sludge. Solids retetion time and temperature were demonstrated to be important parameters affecting the survival of poliovirus, f-2 coliphage, Streptoccus fecalis, and Escherichia coli during anaerobic digestion. The die-off rates of the coliphages were similar to those of the poliovirus at 35/sup 0/C. S. fecalis appeared to be the most stable of any of the bacteria and viruses studied. All organisms were more stable at 25 than at 35/sup 0/C. The data demonstrate that the concentration of enteric bacteria and viruses can be effectively reduced during anaerobic digestion using techniques, such as increased solids retention times and mesophilic temperatures, that are consistent with achieving high methane yields. The survival of enteric viruses during anaerobic digestion may be affected by the characteristics of the feedstock as well as by the process operating conditions.

  9. OCCURRENCE OF ENTERIC VIRUSES IN WATERS

    EPA Science Inventory

    A number of different types of human enteric viruses cause waterborne outbreaks when individuals are exposed to contaminated drinking and recreational waters. Vaccination against poliovirus has virtually eliminated poliomyelitis from the planet, but other members of the enterovi...

  10. OCCURRENCE OF ENTERIC VIRUSES IN SURFACE WATERS

    EPA Science Inventory

    Human enteric viruses cause a number of diseases when individuals are exposed to contaminated drinking & recreational waters. Vaccination against poliovirus has virtually eliminated poliomyelitis from the planet. Other members of enterovirus group cause numerous diseases. Hepatit...

  11. Thermal inactivation of enteric viruses and bioaccumulation of enteric foodborne viruses in live oysters (Crassostrea virginica)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Human enteric viruses are one of the main causative agents of shellfish associated outbreaks. In this study, the kinetics of viral bioaccumulation in live oysters and the heat stability of the most predominant enteric viruses were determined in both tissue culture and in oyster tissues. A human nor...

  12. Removal of Escherichia coli, Enterococcus fecalis, coliphage MS2, poliovirus, and hepatitis A virus from oysters (Crassostrea virginica) and hard shell clams (Mercinaria mercinaria) by depuration.

    PubMed

    Love, David C; Lovelace, Greg L; Sobsey, Mark D

    2010-10-15

    Filter-feeding bivalve mollusks (shellfish) can bioaccumulate pathogenic microorganisms in up to 1000-fold higher levels than overlying waters, and therefore disease risks are associated with consuming raw or partially cooked shellfish. Many of these shellfish-borne diseases are due to enteric bacteria and viruses associated with fecal contamination. To control shellfish-borne diseases, guidelines for shellfish harvest waters and shellfish meat have been devised, which include cleansing of contaminated shellfish by depuration in controlled systems, heat pasteurization, or relay to clean waters. This study examines the depuration of oysters (Crassostrea virginica) and hard shell clams (Mercinaria mercinaria) in a flow-through depuration system under variable temperature (12 °C, 18 °C, and 25 °C), salinity (8 ppt, 18 ppt, and 28 ppt), turbidity (<1NTU, 10NTU, and 20NTU), pH (pH 7 and pH 8), and algae conditions (0 cells/mL and 50,000 cells/mL), with constant dissolved oxygen (5-7 mg/L). Oysters and hard shell clams were artificially contaminated with enteric microorganisms: Escherichia coli, Enterococcus faecalis, coliphage MS2, Poliovirus type-1 and Hepatitis A virus HM-175 (HAV), then depurated in 5-day trials with daily sampling. In oysters, optimizing environmental parameters of water temperature improved E. coli, MS2, poliovirus and HAV depuration, and optimized salinity improved E. coli, E. faecalis, and MS2 depuration rates. In hard shell clams, salinity improved E. coli and E. faecalis depuration rates. Adjusting turbidity, pH or algae did not improve microorganism depuration in either oysters or hard shell clams, with the exception of turbidity on E. faecalis in hard shell clams. Microorganism depuration rates in oysters from greatest to least were: MS2>E. coli>E. faecalis>poliovirus>HAV, and in clams depuration rates from greatest to least were: E. coli>E. faecalis>HAV>MS2>poliovirus. Because E. coli and E. faecalis were removed at faster rates than HAV

  13. Somatic Coliphage Profiles of Produce and Environmental Samples from Farms in Northern México.

    PubMed

    Bartz, Faith E; Hodge, Domonique Watson; Heredia, Norma; de Aceituno, Anna Fabiszewski; Solís, Luisa; Jaykus, Lee-Ann; Garcia, Santos; Leon, Juan S

    2016-09-01

    Somatic coliphages were quantified in 459 produce and environmental samples from 11 farms in Northern Mexico to compare amounts of somatic coliphages among different types of fresh produce and environmental samples across the production steps on farms. Rinsates from cantaloupe melons, jalapeño peppers, tomatoes, and the hands of workers, soil, and water were collected during 2011-2012 at four successive steps on each farm, from the field before harvest through the packing facility, and assayed by FastPhage MPN Quanti-tray method. Cantaloupe farm samples contained more coliphages than jalapeño or tomato (p range <0.01-0.03). Across production steps, jalapeños had higher coliphage percentages before harvest than during packing (p = 0.03), while tomatoes had higher coliphage concentrations at packing than all preceding production steps (p range <0.01-0.02). These findings support the use of targeted produce-specific interventions at multiple points in the process of growing and packing produce to reduce the risk of enteric virus contamination and improve food safety during fruit and vegetable production.

  14. Field-based evaluation of a male-specific (F+) RNA coliphage ...

    EPA Pesticide Factsheets

    Fecal contamination of water poses a significant risk to public health due to the potential presence of pathogens, including enteric viruses. Thus, sensitive, reliable and easy to use methods for the detection of microorganisms are needed to evaluate water quality. In this study, we performed a field evaluation of an anion-exchange resin based platform to concentrate F-RNA coliphages (fecal/enteric virus indicators) from diverse fecally impacted environmental waters. In this platform, F-RNA coliphages are adsorbed to anion-exchange resin and direct nucleic acid isolation is performed, yielding a sample amenable to real-time reverse transcriptase PCR detection. Matrix-dependent inhibition was evaluated using known quantities of spiked F-RNA coliphage genogroups GI, GII, GII and GIV. Detection was successful in 97%, 72%, 85% and 98% of the samples for spiked F-RNA coliphage GI, GII, GIII and GIV, respectively, and was differentially affected by inhibitory properties specific to each water sample. No association between inhibition and the water samples’ physicochemical properties was apparent. Parallel evaluations of the spiked samples with internal amplification control (IAC) reactions (a widely used control to assess inhibition) demonstrated that IAC reaction inhibition was not agreement with that observed for spiked samples, suggesting that testing of spiked samples allows for better assessments of matrix-dependent inhibition. Additionally, the anion-

  15. Interaction of human adenoviruses and coliphages with kaolinite and bentonite.

    PubMed

    Bellou, Maria I; Syngouna, Vasiliki I; Tselepi, Maria A; Kokkinos, Petros A; Paparrodopoulos, Spyros C; Vantarakis, Apostolos; Chrysikopoulos, Constantinos V

    2015-06-01

    Human adenoviruses (hAdVs) are pathogenic viruses responsible for public health problems worldwide. They have also been used as viral indicators in environmental systems. Coliphages (e.g., MS2, ΦX174) have also been studied as indicators of viral pollution in fecally contaminated water. Our objective was to evaluate the distribution of three viral fecal indicators (hAdVs, MS2, and ΦΧ174), between two different phyllosilicate clays (kaolinite and bentonite) and the aqueous phase. A series of static and dynamic experiments were conducted under two different temperatures (4, 25°C) for a time period of seven days. HAdV adsorption was examined in DNase I reaction buffer (pH=7.6, and ionic strength (IS)=1.4mM), whereas coliphage adsorption in phosphate buffered saline solution (pH=7, IS=2mM). Moreover, the effect of IS on hAdV adsorption under static conditions was evaluated. The adsorption of hAdV was assessed by real-time PCR and its infectivity was tested by cultivation methods. The coliphages MS2 and ΦΧ174 were assayed by the double-layer overlay method. The experimental results have shown that coliphage adsorption onto both kaolinite and bentonite was higher for the dynamic than the static experiments; whereas hAdV adsorption was lower under dynamic conditions. The adsorption of hAdV increased with decreasing temperature, contrary to the results obtained for the coliphages. This study examines the combined effect of temperature, agitation, clay type, and IS on hAdV adsorption onto clays. The results provide useful new information on the effective removal of viral fecal indicators (MS2, ΦX174 and hAdV) from dilute aqueous solutions by adsorption onto kaolinite and bentonite. Factors enabling enteric viruses to penetrate soils, groundwater and travel long distances within aquifers are important public health issues. Because the observed adsorption behavior of surrogate coliphages MS2 and ΦΧ174 is substantially different to that of hAdV, neither MS2 nor

  16. Incidence of Enteric Viruses in Groundwater from Household Wells in Wisconsin

    PubMed Central

    Borchardt, Mark A.; Bertz, Phil D.; Spencer, Susan K.; Battigelli, David A.

    2003-01-01

    Recent studies on the contamination of groundwater with human enteric viruses have focused on public water systems, whereas little is known about the occurrence of viruses in private household wells. The objective of the present study was to estimate the incidence of viruses in Wisconsin household wells located near septage land application sites or in rural subdivisions served by septic systems. Fifty wells in seven hydrogeologic districts were sampled four times over a year, once each season. Reverse transcriptase PCR (RT-PCR), followed by Southern hybridization, was used to detect enteroviruses, rotavirus, hepatitis A virus (HAV), and Norwalk-like viruses (NLVs). In addition, cell culture was used to detect culturable enteroviruses. Companion water samples were collected for total coliforms, Escherichia coli, fecal enterococci, F-specific RNA coliphages, nitrate, and chloride analyses. Among the 50 wells, four (8%) were positive for viruses by RT-PCR. Three wells were positive for HAV, and the fourth well was positive for both rotavirus and NLV in one sample and an enterovirus in another sample. Contamination was transient, since none of the wells was virus positive for two sequential samples. Culturable enteroviruses were not detected in any of the wells. Water quality indicators were not statistically associated with virus occurrence, although some concordance was noted for chloride. The present study is the first in the United States to systematically monitor private household wells for virus contamination and, combined with data for public wells, provides further insight on the extent of groundwater contamination with human enteric viruses. PMID:12571044

  17. 9 CFR 113.204 - Mink Enteritis Vaccine, Killed Virus.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Mink Enteritis Vaccine, Killed Virus... REQUIREMENTS Killed Virus Vaccines § 113.204 Mink Enteritis Vaccine, Killed Virus. Mink Enteritis Vaccine... prior to challenge. If unfavorable reactions attributable to the vaccine occur, the serial...

  18. 9 CFR 113.204 - Mink Enteritis Vaccine, Killed Virus.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Mink Enteritis Vaccine, Killed Virus... REQUIREMENTS Killed Virus Vaccines § 113.204 Mink Enteritis Vaccine, Killed Virus. Mink Enteritis Vaccine... prior to challenge. If unfavorable reactions attributable to the vaccine occur, the serial...

  19. 9 CFR 113.204 - Mink Enteritis Vaccine, Killed Virus.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Mink Enteritis Vaccine, Killed Virus... REQUIREMENTS Killed Virus Vaccines § 113.204 Mink Enteritis Vaccine, Killed Virus. Mink Enteritis Vaccine... prior to challenge. If unfavorable reactions attributable to the vaccine occur, the serial...

  20. 9 CFR 113.204 - Mink Enteritis Vaccine, Killed Virus.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Mink Enteritis Vaccine, Killed Virus... REQUIREMENTS Killed Virus Vaccines § 113.204 Mink Enteritis Vaccine, Killed Virus. Mink Enteritis Vaccine... prior to challenge. If unfavorable reactions attributable to the vaccine occur, the serial...

  1. 9 CFR 113.204 - Mink Enteritis Vaccine, Killed Virus.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Mink Enteritis Vaccine, Killed Virus... REQUIREMENTS Killed Virus Vaccines § 113.204 Mink Enteritis Vaccine, Killed Virus. Mink Enteritis Vaccine... prior to challenge. If unfavorable reactions attributable to the vaccine occur, the serial...

  2. Thermal Inactivation of Enteric Viruses and Bioaccumulation of Enteric Foodborne Viruses in Live Oysters (Crassostrea virginica)

    PubMed Central

    Araud, Elbashir; DiCaprio, Erin; Ma, Yuanmei; Lou, Fangfei; Gao, Yu; Kingsley, David; Hughes, John H.

    2016-01-01

    Human enteric viruses are among the main causative agents of shellfish-associated outbreaks. In this study, the kinetics of viral bioaccumulation in live oysters and the heat stabilities of the predominant enteric viruses were determined both in tissue culture and in oyster tissues. A human norovirus (HuNoV) GII.4 strain, HuNoV surrogates (murine norovirus [MNV-1], Tulane virus [TV]), hepatitis A virus (HAV), and human rotavirus (RV) bioaccumulated to high titers within oyster tissues, with different patterns of bioaccumulation for the different viruses. We tested the thermal stability of each virus at 62, 72, and 80°C in culture medium. The viruses can be ranked from the most heat resistant to the least stable as follows: HAV, RV, TV, MNV-1. In addition, we found that oyster tissues provided protection to the viruses during heat treatment. To decipher the mechanism underlying viral inactivation by heat, purified TV was treated at 80°C for increasing time intervals. It was found that the integrity of the viral capsid was disrupted, whereas viral genomic RNA remained intact. Interestingly, heat treatment leading to complete loss of TV infectivity was not sufficient to completely disrupt the receptor binding activity of TV, as determined by the porcine gastric mucin–magnetic bead binding assay. Similarly, HuNoV virus-like particles (VLPs) and a HuNoV GII.4 strain retained some receptor binding ability following heat treatment. Although foodborne viruses have variable heat stability, 80°C for >6 min was sufficient to completely inactivate enteric viruses in oysters, with the exception of HAV. PMID:26826225

  3. 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.

  4. 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

  5. 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.

  6. Development of reference antisera to enteric-origin avian viruses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent molecular surveys have revealed geographically distinct lineages of avian reovirus, rotavirus and astrovirus circulating in commercial poultry. To improve our understanding of enteric virus pathogenesis, specific immunological reagents are needed to detect viruses in histological samples. To ...

  7. Enteric virus and vibrio contamination of shellfish: intervention strategies

    Technology Transfer Automated Retrieval System (TEKTRAN)

    INTRODUCTION. Molluscan shellfish include oysters, clams, mussels, and cockles, which can cause illnesses from a variety of human pathogens. Enteric viruses, like norovirus and hepatitis A virus, are generally transmitted to shellfish through fecal contamination of shellfish harvesting areas, alth...

  8. Enteric viruses of chickens and turkeys

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Although enteric disease in commercial poultry operations is common, and often unofficially reported and discussed by field veterinarians as “non-specific enteric disease”, three recognized enteric syndromes do exist in poultry: poult enteritis complex (PEC) and poult enteritis mortality syndrome (P...

  9. Factors affecting decay of Salmonella Birkenhead and coliphage MS2 during mesophilic anaerobic digestion and air drying of sewage sludge.

    PubMed

    Mondal, Tania; Rouch, Duncan A; Thurbon, Nerida; Smith, Stephen R; Deighton, Margaret A

    2015-06-01

    Factors affecting the decay of Salmonella Birkenhead and coliphage, as representatives of bacterial and viral pathogens, respectively, during mesophilic anaerobic digestion (MAD) and air drying treatment of anaerobically digested sewage sludge were investigated. Controlled concentrations of S. Birkenhead were inoculated into non-sterile, autoclaved, γ-irradiated and nutrient-supplemented sludge and cultures were incubated at 37 °C (MAD sludge treatment temperature) or 20 °C (summer air drying sludge treatment temperature). Nutrient limitation caused by microbial competition was the principal mechanism responsible for the decay of S. Birkenhead by MAD and during air drying of digested sludge. The effects of protease activity in sludge on MS2 coliphage decay in digested and air dried sludge were also investigated. MS2 coliphage showed a 3.0-3.5 log10 reduction during incubation with sludge-protease extracts at 37 °C for 25 h. Proteases produced by indigenous microbes in sludge potentially increase coliphage inactivation and may therefore have a significant role in the decay of enteric viruses in sewage sludge. The results help to explain the loss of viability of enteric bacteria and viral pathogens with treatment process time and contribute to fundamental understanding of the various biotic inactivation mechanisms operating in sludge treatment processes at mesophilic and ambient temperatures.

  10. Processing Strategies to Inactivate Enteric Viruses in Shellfish: Limitations of Surrogate Viruses and Molecular Methods

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Noroviruses, hepatitis A and E viruses, sapovirus, astrovirus, rotavirus, Aichi virus, enteric adenoviruses, poliovirus, and other enteroviruses enter shellfish through contaminated seawater or by contamination during handling and processing, resulting in outbreaks ranging from isolated to epidemic....

  11. Replication of coliphage Q beta as affected by host cell number, nutrition, competition from insusceptible cells and non-FRNA coliphages.

    PubMed

    Woody, M A; Cliver, D O

    1997-04-01

    F-specific RNA (FRNA) coliphages, which infect Escherichia coli by attachment to F pili, might serve as indicators of human enteric viruses in groundwater, provided these phages do not replicate in groundwater and replicate only to a limited extent in wastewater. Several factors that could influence phage replication in either of these environments were examined. Q beta did not replicate when host cells were fewer than 10(4) cfu ml-1. Replication selected for insusceptible cells when Q beta was incubated with its E. coli host. Loss of Q beta, presumably by inactivation, occurred in autoclaved on-site and urban wastewater, autoclaved groundwater, and in filter-sterilized spent LB broth. Replication did not occur in LB broth diluted with sterile saline to 1% of its original strength, which indicates that replication of FRNA coliphages cannot occur in such nutrient-poor environments as wastewater and groundwater. Competition from non-FRNA coliphages and insusceptible cells tended to reduce Q beta replication, as predicted, but phage yields unexpectedly increased significantly when Enterococcus faecalis was added to cultures.

  12. Persistence of enteric viruses within oysters (Crassostrea virginica)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    It is well known that water-borne enteric viruses are concentrated by bivalves. Why these viruses are selectively retained and remain infectious within shellfish tissues for extended periods is unknown. Our current hypothesis is that phagocytic hemocytes (blood cells) are a site of virus persiste...

  13. AN OVERVIEW OF ENTERIC VIRUS EXTRACTION AND ASSAY METHODS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Enteric viruses, particularly norovirus and hepatitis A virus, are major contaminants of molluscan shellfish, leading to outbreaks of viral illness. A host of procedures have been developed for the extraction and assay of viruses from shellfish. Early extraction and assay methods focused on the de...

  14. 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

  15. Survival of human enteric viruses in the environment and food.

    PubMed

    Rzezutka, Artur; Cook, Nigel

    2004-10-01

    Human enteric pathogenic viruses can enter the environment through discharge of waste materials from infected persons, and be transmitted back to susceptible persons to continue the cycle of disease. Contamination of food with viruses may also promote disease outbreaks. A number of studies have investigated the survival characteristics of several enteric viruses in various environments and foodstuffs, to help explain the transmissibility of these pathogens. This review deals with published work on enteric virus survival on fomites, and in waters, soil, and foods; the results of these studies have illustrated the robust survival of viruses in these environments. Much information is lacking, however, especially for foodstuffs and soils, and no detailed information is available concerning the survival of noroviruses, the most significant foodborne type.

  16. Field survey of enteric viruses in solid waste landfill leachates.

    PubMed Central

    Sobsey, M D

    1978-01-01

    Because municipal solid waste may contain fecal material from a variety of sources, there is concern that the leachate discharged from some solid waste landfills may contain enteric pathogens, including enteric viruses. In this study, 22 leachate samples from 21 different landfills in the United States and Canada were examined for enteric viruses. The sites represented a broad range of conditions for solid waste landfills and the leachate samples ranged from 10.3 to 18 liters in volume. Enteric viruses were found in only one of the 22 leachate samples examined. Two viruses, identified as poliovirus types 1 and 3, were found in an 11.8 liter sample obtained from a site where solid waste landfill practice was deficient. The low levels of enteric viruses detected in field samples of raw leachate and the opportunities for further reductions in the virus concentration of leachates by such processes as thermal inactivation, removal by soil and dilution in ground and surface waters, suggest that leachates from properly operated solid waste landfills do not constitute an environmental or public health hazard due to enteric viruses. PMID:28677

  17. Detection of enteric viruses in shellfish

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Norovirus and hepatitis A virus contamination are significant threats to the safety of shellfish and other foods. Methods for the extraction and assay of these viruses from shellfish are complex, time consuming, and technically challenging. Here, we itemize some of the salient points in extracting...

  18. Presence of human enteric viruses in deep urban groundwater

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Enteric viruses, because of their small size, have a high potential to move deeply through the subsurface environment, penetrate aquitards, and reach confined aquifers. Until recently, few water utilities or researchers were aware of possible virus presence in deep aquifers and wells. Over the past ...

  19. A dilemma for viruses and giant viruses: which endocytic pathway to use to enter cells?

    PubMed

    Ghigo, Eric

    2010-01-01

    Viruses must enter host cells to deliver their genetic material and accessory proteins. Endocytosis offers to viruses the opportunity to enter host cells. However, endocytosis is a complex phenomenon that includes different mechanisms, clathrin-mediated endocytosis, caveolin-mediated endocytosis, macropinocytosis, and phagocytosis. Here, I describe the ways used by different viruses to exploit these endocytic pathways.

  20. Identification of Enteric Viruses in Foods from Mexico City.

    PubMed

    Parada-Fabián, José Carlos; Juárez-García, Patricia; Natividad-Bonifacio, Iván; Vázquez-Salinas, Carlos; Quiñones-Ramírez, Elsa Irma

    2016-09-01

    Foodborne viruses are a common and, probably, the most under-recognized cause of outbreaks of gastroenteritis. Among the main foods involved in the transmission of human enteric viruses are mollusks, and fruits and vegetables irrigated with wastewater and/or washed with non-potable water or contaminated by contact with surfaces or hands of the infected personnel during its preparation. In this study, 134 food samples were analyzed for the detection of Norovirus, Rotavirus, and Hepatitis A virus (HAV) by amplification of conserved regions of these viruses. From the 134 analyzed samples, 14 were positive for HAV, 6 for Norovirus, and 11 for Rotavirus. This is the first report in Mexico where emphasis is given to the presence of HAV and Norovirus on perishable foods and food from fisheries, as well as Rotavirus on frozen vegetables, confirming the role of vegetables and bivalve mollusks as transmitting vehicles of enteric viruses.

  1. Phytocompounds for the control of human enteric viruses.

    PubMed

    D'Souza, Doris H

    2014-02-01

    Plant extracts and associated polyphenols are known for their varied health benefits that include antioxidant effects and antimicrobial properties. The increasing consumer demand for cost-effective and natural alternatives to chemically-synthesized antimicrobials and therapeutics that are also sustainable makes the field of phytochemical research rather intriguing and challenging. Human enteric viruses are increasingly recognized worldwide as significant causes of human disease in adults and children, alike. In the absence of available vaccines for the human noroviruses, plant extracts are gaining popularity for the prevention and treatment of viral diseases. Research on plant extracts (particularly polyphenols derived from fruits) for human enteric virus control will be briefly summarized in this article.

  2. Enteric and indicator virus removal by surface flow wetlands.

    PubMed

    Rachmadi, Andri T; Kitajima, Masaaki; Pepper, Ian L; Gerba, Charles P

    2016-01-15

    We investigated the occurrence and attenuation of several human enteric viruses (i.e., norovirus, adenovirus, Aichi virus 1, polyomaviruses, and enterovirus) as well as a plant virus, pepper mild mottle virus (PMMoV), at two surface flow wetlands in Arizona. The retention time in one of the wetlands was seven days, whereas in the other wetland it could not be defined. Water samples were collected at the inlet and outlet from the wetlands over nine months, and concentration of viral genomes was determined by quantitative polymerase chain reaction (qPCR). Of the human enteric viruses tested, adenovirus and Aichi virus 1 were found in the greatest prevalence in treated wastewater (i.e., inlet of the wetlands). Reduction efficiencies of enteric viruses by the wetlands ranged from 1 to 3 log10. Polyomaviruses were generally removed to below detection limit, indicating at least 2 to 4 log10 removal. PMMoV was detected in a greater concentration in the inlet of both wetlands for all the viruses tested (10(4) to 10(7) genome copies/L), but exhibited little or no removal (1 log10 or less). To determine the factors associated with virus genome attenuation (as determined by qPCR), the persistence of PMMoV and poliovirus type 1 (an enterovirus) was studied in autoclaved and natural wetland water, and deionized water incubated under three different temperatures for 21 days. A combination of elevated water temperature and biological activities reduced poliovirus by 1 to 4 log10, while PMMoV was not significantly reduced during this time period. Overall, PMMoV showed much greater persistence than human viruses in the wetland treatment.

  3. The effect of carvacrol on enteric viruses.

    PubMed

    Sánchez, C; Aznar, R; Sánchez, G

    2015-01-02

    Carvacrol, a monoterpenic phenol, is said to have extensive antimicrobial activity in a wide range of food spoilage or pathogenic fungi, yeast and bacteria. The aim of this study was to assess its antiviral activity on norovirus surrogates, feline calicivirus (FCV), murine norovirus (MNV), and hepatitis A virus (HAV), as well as its potential in food applications. Initially, different concentrations of carvacrol (0.25, 0.5, 1%) were individually mixed with each virus at titers of ca. 6-7 log TCID50/ml and incubated 2h at 37°C. Carvacrol at 0.5% completely inactivated the two norovirus surrogates, whereas 1% concentration was required to achieve ca. 1 log reduction of HAV. In lettuce wash water, carvacrol efficacy on MNV was dependent on the chemical oxygen demand (COD), with no effect over 300 ppm. A 4 log reduction in FCV infectivity was observed when 0.5% carvacrol was used to sanitize lettuce wash water, regardless of COD. Carvacrol was also evaluated as a natural disinfectant of produce, showing 1% carvacrol reduced inoculated NoV surrogates titers in lettuce by 1 log after 30 min contact. These results represent a step forward in improving food safety by using carvacrol as an alternative natural additive to reduce viral contamination in the fresh vegetable industry.

  4. Expression and serological reactivity of hemorrhagic enteritis virus hexon protein.

    PubMed

    Lobová, Dana; Celer, Vladimír

    2016-05-01

    The aim of this work was to express the recombinant hexon protein of the hemorrhagic enteritis virus, to establish the diagnostic value of this protein for serological detection of antibodies in turkey serum samples and to assess seroprevalence of the infection in the Czech Republic. The N' terminal part of the hexon protein was expressed in a bacterial expression system and used as an antigen in an ELISA test for the detection of hemorrhagic enteritis virus specific antibodies in turkey sera. Validation of the test was performed by comparison with a commercially available ELISA test. Serological reactivity was assessed on a panel of 126 turkey sera by a newly developed ELISA test. Serum samples were taken from turkey farms with the history of hemorrhagic enteritis virus infection, from farms with animals free of infection, and from turkey farms following vaccination. Both ELISA kits gave identical results (100 %) with the tested sera. ELISA based on the recombinant hexon protein thus proved useful and cheaper for detection of antibodies in turkey flocks infected with the hemorrhagic enteritis virus.

  5. Enteritis

    MedlinePlus

    ... enteritis E coli enteritis Food poisoning Radiation enteritis Salmonella enteritis Shigella enteritis Staph aureus food poisoning Symptoms ... store food that needs to stay chilled. Images Salmonella typhi organism Yersinia enterocolitica organism Campylobacter jejuni organism ...

  6. Coliphages as ecological indicators of enteroviruses in various water systems*

    PubMed Central

    Šimková, A.; Červenka, J.

    1981-01-01

    The occurrence of coliphages and enteroviruses in a variety of water systems in Czechoslovakia was monitored for two years. Two host strains of Escherichia coli bacteria were used to test 1161 water samples for the presence of bacteriophages. These strains were polyvalent hosts for a broad spectrum of morphologically distinct coliphages, and their use thus gave quantitative data on the degree of viral pollution in any given water sample. Ninety-two water samples were tested in parallel for the presence of enteroviruses, by using a flocculation method to concentrate the viruses followed by isolation in cultures of a buffalo green monkey (BGM) kidney continuous cell line. The enterovirus and coliphage recovery rates showed similar differences when waters with different levels of pollution were compared. Seasonal fluctuations of both the coliphage and enterovirus (mostly poliovirus) levels in river water were demonstrated by statistical analysis of the data collected. The levels increased in the winter and sharply declined in the summer months as the river water temperature increased. Chemical pollution did not seem to influence the survival of either the coliphages or the enteroviruses in the observed rivers. PMID:6274536

  7. Hemocytes Are Sites of Enteric Virus Persistence within Oysters ▿

    PubMed Central

    Provost, Keleigh; Dancho, Brooke A.; Ozbay, Gulnihal; Anderson, Robert S.; Richards, Gary P.; Kingsley, David H.

    2011-01-01

    The goal of this study was to determine how enteric viruses persist within shellfish tissues. Several lines of novel evidence show that phagocytic blood cells (hemocytes) of Eastern oysters (Crassostrea virginica) play an important role in the retention of virus particles. Our results demonstrated an association of virus contamination with hemocytes but not with hemolymph. Live oysters contaminated overnight with hepatitis A virus (HAV) and murine norovirus (MNV) had 56% and 80% of extractable virus associated with hemocytes, respectively. Transfer of HAV-contaminated hemocytes to naïve (virus-free) oysters resulted in naïve oyster meat testing HAV positive for up to 3 weeks. Acid tolerance of HAV, MNV, poliovirus (PV), and feline calicivirus (FCV) correlated with the ability of each virus to persist within oysters. Using reverse transcription-PCR (RT-PCR) to evaluate persistence of these viruses in oysters, we showed that HAV persisted the longest (>21 days) and was most acid resistant, MNV and PV were less tolerant of acidic pH, persisting for up to 12 days and 1 day, respectively, and FCV did not persist (<1 day) within oysters and was not acid tolerant. This suggests that the ability of a virus to tolerate the acidic conditions typical of phagolysosomal vesicles within hemocytes plays a role in determining virus persistence in shellfish. Evaluating oyster and hemocyte homogenates and live contaminated oysters as a prelude to developing improved viral RNA extraction methods, we found that viruses were extracted more expediently from hemocytes than from whole shellfish tissues and gave similar RT-PCR detection sensitivities. PMID:21948840

  8. Detection of enteric viruses in treated drinking water.

    PubMed Central

    Keswick, B H; Gerba, C P; DuPont, H L; Rose, J B

    1984-01-01

    The occurrence of viruses in conventionally treated drinking water derived from a heavily polluted source was evaluated by collecting and analyzing 38 large-volume (65- to 756-liter) samples of water from a 9 m3/s (205 X 10(6) gallons [776 X 10(6) liters] per day) water treatment plant. Samples of raw, clarified, filtered, and chlorinated finished water were concentrated by using the filter adsorption-elution technique. Of 23 samples of finished water, 19 (83%) contained viruses. None of the nine finished water samples collected during the dry season contained detectable total coliform bacteria. Seven of nine finished water samples collected during the dry season met turbidity, total coliform bacteria, and total residual chlorine standards. Of these, four contained virus. During the dry season the percent removals were 25 to 93% for enteric viruses, 89 to 100% for bacteria, and 81% for turbidity. During the rainy season the percent removals were 0 to 43% for enteric viruses, 80 to 96% for bacteria, and 63% for turbidity. None of the 14 finished water samples collected during the rainy season met turbidity standards, and all contained rotaviruses or enteroviruses. PMID:6331313

  9. Enteric viruses in a mangrove lagoon, survival and shellfish incidence

    SciTech Connect

    Lopez de Cardona, I.; Bermudez, M.; Billmire, E.; Hazen, T.C.

    1988-12-31

    Mangrove oysters (Crassostrea rhizophorae) were screened for enteric viruses. For 18 months oysters were collected from Cano Boqueron, a tropical mangrove lagoon on the southwest coast of Puerto Rico. This popular tourist resort has two primary sewage treatment plants which service 158 single family cabanas. In spite of the heavy seasonal input of sewage to Cano Boqueron and high densities of fecal coliform bacteria, enteric viruses were not detected in shellfish meat. Because no viruses were detected in the oysters, a virus survival study was performed. Poliovirus type 1 was placed in diffusion chambers in situ at two sites in Cano Boqueron. More than 95% of the poliovirus inactivation occurred within 24 h. Virus inactivation was significantly different by site, indicating different inactivation rates within the lagoon. Chamber studies done simultaneously with Escherichia coli did not reveal differences between sites. It is suggested that the sewage effluent had an antiviral effect in the absence of an antibacterial effect. This study demonstrates the importance for establishing microbial contamination standards for shellfish growing waters in the tropics based upon in situ studies with tropical species, e.g. mangrove oyster.

  10. Inactivation of enteric viruses in minimally processed berries and herbs.

    PubMed

    Butot, S; Putallaz, T; Amoroso, R; Sánchez, G

    2009-06-01

    Several hepatitis A virus (HAV) and human norovirus (HuNoV) outbreaks due to consumption of contaminated berries and vegetables have recently been reported. Model experiments were performed to determine the effectiveness of freeze-drying, freeze-drying combined with heating, and steam blanching for inactivation of enteric viruses that might be present on the surface of berries and herbs. Inactivation of HAV and inactivation of feline calicivirus, a surrogate for HuNoV, were assessed by viral culturing and quantitative reverse transcription PCR (RT-PCR), whereas HuNoV survival was determined only by quantitative RT-PCR. While freeze-drying barely reduced (<1.3 log(10) units) the amount of HAV RNA detected in frozen produce, a greater decline in HAV infectivity was observed. The resistance of HuNoV genogroup I (GI) to freeze-drying was significantly higher than that of HuNoV GII on berries. Addition of a terminal dry heat treatment at 120 degrees C after freeze-drying enhanced virus inactivation by at least 2 log(10) units, except for HuNoV GII. The results suggest that steam blanching at 95 degrees C for 2.5 min effectively inactivated infectious enteric viruses if they were present in herbs. Our results provide data for adjusting food processing technologies if viral contamination of raw materials is suspected.

  11. Coliphages as indicators of gastrointestinal illness in recreational waters: a pooled analysis of six prospective marine beach cohorts

    EPA Science Inventory

    Background: Coliphages have been proposed as potential indicators of fecal contamination of marine recreational waters because they may better predict the presence of viruses than fecal indicator bacteria. We estimated the association between the presence of coliphages and self-r...

  12. Norovirus and other human enteric viruses in moroccan shellfish.

    PubMed

    Benabbes, Laila; Ollivier, Joanna; Schaeffer, Julien; Parnaudeau, Sylvain; Rhaissi, Houria; Nourlil, Jalal; Le Guyader, Françoise S

    2013-03-01

    The aim of this study was to evaluate the presence of human enteric viruses in shellfish collected along the Mediterranean Sea and Atlantic Coast of Morocco. A total of 77 samples were collected from areas potentially contaminated by human sewage. Noroviruses were detected in 30 % of samples, with an equal representation of GI and GII strains, but were much more frequently found in cockles or clams than in oysters. The method used, including extraction efficiency controls, allowed the quantification of virus concentration. As in previous reports, results showed levels of contamination between 100 and 1,000 copies/g of digestive tissues. Sapoviruses were detected in 13 % of samples mainly in oyster and clam samples. Hepatitis A virus was detected in two samples, with concentrations around 100 RNA copies/g of digestive tissues. Only two samples were contaminated with enterovirus and none with norovirus GIV or Aichi virus. This study highlights the interest of studying shellfish samples from different countries and different production areas. A better knowledge of shellfish contamination helps us to understand virus levels in shellfish and to improve shellfish safety, thus protecting consumers.

  13. African Swine Fever Virus Uses Macropinocytosis to Enter Host Cells

    PubMed Central

    Sánchez, Elena G.; Quintas, Ana; Pérez-Núñez, Daniel; Nogal, Marisa; Barroso, Susana; Carrascosa, Ángel L.; Revilla, Yolanda

    2012-01-01

    African swine fever (ASF) is caused by a large and highly pathogenic DNA virus, African swine fever virus (ASFV), which provokes severe economic losses and expansion threats. Presently, no specific protection or vaccine against ASF is available, despite the high hazard that the continued occurrence of the disease in sub-Saharan Africa, the recent outbreak in the Caucasus in 2007, and the potential dissemination to neighboring countries, represents. Although virus entry is a remarkable target for the development of protection tools, knowledge of the ASFV entry mechanism is still very limited. Whereas early studies have proposed that the virus enters cells through receptor-mediated endocytosis, the specific mechanism used by ASFV remains uncertain. Here we used the ASFV virulent isolate Ba71, adapted to grow in Vero cells (Ba71V), and the virulent strain E70 to demonstrate that entry and internalization of ASFV includes most of the features of macropinocytosis. By a combination of optical and electron microscopy, we show that the virus causes cytoplasm membrane perturbation, blebbing and ruffles. We have also found that internalization of the virions depends on actin reorganization, activity of Na+/H+ exchangers, and signaling events typical of the macropinocytic mechanism of endocytosis. The entry of virus into cells appears to directly stimulate dextran uptake, actin polarization and EGFR, PI3K-Akt, Pak1 and Rac1 activation. Inhibition of these key regulators of macropinocytosis, as well as treatment with the drug EIPA, results in a considerable decrease in ASFV entry and infection. In conclusion, this study identifies for the first time the whole pathway for ASFV entry, including the key cellular factors required for the uptake of the virus and the cell signaling involved. PMID:22719252

  14. African swine fever virus uses macropinocytosis to enter host cells.

    PubMed

    Sánchez, Elena G; Quintas, Ana; Pérez-Núñez, Daniel; Nogal, Marisa; Barroso, Susana; Carrascosa, Ángel L; Revilla, Yolanda

    2012-01-01

    African swine fever (ASF) is caused by a large and highly pathogenic DNA virus, African swine fever virus (ASFV), which provokes severe economic losses and expansion threats. Presently, no specific protection or vaccine against ASF is available, despite the high hazard that the continued occurrence of the disease in sub-Saharan Africa, the recent outbreak in the Caucasus in 2007, and the potential dissemination to neighboring countries, represents. Although virus entry is a remarkable target for the development of protection tools, knowledge of the ASFV entry mechanism is still very limited. Whereas early studies have proposed that the virus enters cells through receptor-mediated endocytosis, the specific mechanism used by ASFV remains uncertain. Here we used the ASFV virulent isolate Ba71, adapted to grow in Vero cells (Ba71V), and the virulent strain E70 to demonstrate that entry and internalization of ASFV includes most of the features of macropinocytosis. By a combination of optical and electron microscopy, we show that the virus causes cytoplasm membrane perturbation, blebbing and ruffles. We have also found that internalization of the virions depends on actin reorganization, activity of Na(+)/H(+) exchangers, and signaling events typical of the macropinocytic mechanism of endocytosis. The entry of virus into cells appears to directly stimulate dextran uptake, actin polarization and EGFR, PI3K-Akt, Pak1 and Rac1 activation. Inhibition of these key regulators of macropinocytosis, as well as treatment with the drug EIPA, results in a considerable decrease in ASFV entry and infection. In conclusion, this study identifies for the first time the whole pathway for ASFV entry, including the key cellular factors required for the uptake of the virus and the cell signaling involved.

  15. Comparison of canine parvovirus with mink enteritis virus by restriction site mapping.

    PubMed Central

    McMaster, G K; Tratschin, J D; Siegl, G

    1981-01-01

    The genomes of canine parvovirus and mink enteritis virus were compared by restriction enzyme analysis of their replicative-form DNAs. Of 79 mapped sites, 68, or 86%, were found to be common for both types of DNA, indicating that canine parvovirus and mink enteritis virus are closely related viruses. Whether they evolved from a common precursor or whether canine parvovirus is derived from mink enteritis virus, however, cannot be deduced from our present data. Images PMID:6264109

  16. Molecular detection of enteric viruses from diarrheic calves in Egypt.

    PubMed

    Mohamed, Fakry F; Mansour, Shimaa M G; El-Araby, Iman E; Mor, Sunil K; Goyal, Sagar M

    2017-01-01

    Neonatal calf diarrhea (NCD) is a major cause of morbidity, mortality and economic losses in the beef and dairy industries. This study was conducted to investigate the existence of enteric viruses in two Egyptian farms with a history of recurrent diarrhea. Fecal samples were collected from 25 diarrheic calves. RNA was extracted and tested by reverse transcription polymerase chain reaction (RT-PCR) for the presence of rotavirus, norovirus, astrovirus, torovirus, coronavirus and bovine viral diarrhea virus. Overall, 76 % (19/25) of samples tested positive for one or more viruses. Rota-, noro- and astroviruses were detected in 48 %, 24 % and 32 % of tested samples, respectively. About 37 % (7/19) of positive samples had two different viruses. One-month-old calves were the group most vulnerable to infections. Based on phylogenetic analysis, bovine rotaviruses were of genotypes G6 and G10, bovine noroviruses were in GIII.2, and bovine astroviruses were in the BAstV lineage 1. Astrovirus sequences showed a high level nucleotide sequence similarity with the Brazilian BAstV sequences available in GenBank. We believe this is the first report of bovine norovirus and bovine astrovirus circulating among calves in Egypt. Further epidemiological studies are recommended to investigate their presence on a wider scale, to predict their association with NCD, and to design appropriate diagnostic and control methods.

  17. Shellfish-associated enteric virus illness: virus localization, disease outbreaks and prevention

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Numerous outbreaks of shellfish-borne enteric virus illness have been reported worldwide. Most notable among the outbreaks are those involving norovirus illness and hepatitis A. Lessons learned from outbreak investigations indicate that most outbreaks are preventable. Anthropogenic sources of con...

  18. The influence of commensal bacteria on infection with enteric viruses

    PubMed Central

    Karst, Stephanie M.

    2016-01-01

    The intestinal microbiota exerts a marked influence in the mammalian host, both during homeostasis and disease. However, until very recently, there has been relatively little focus on the potential effect of commensal microorganisms on viral infection of the intestinal tract. In this Progress article, I review the recent advances that elucidate the mechanisms by which enteric viruses use commensal bacteria to enhance viral infectivity. These mechanisms segregate into two general categories: the direct facilitation of viral infection, including bacterial stabilization of viral particles and the facilitation of viral attachment to host target cells; and the indirect skewing of the antiviral immune response in a manner that promotes viral infection. Finally, I discuss the implications of these interactions for the development of vaccines and novel therapeutic approaches. PMID:26853118

  19. The influence of commensal bacteria on infection with enteric viruses.

    PubMed

    Karst, Stephanie M

    2016-04-01

    The intestinal microbiota exerts a marked influence in the mammalian host, both during homeostasis and disease. However, until very recently, there has been relatively little focus on the potential effect of commensal microorganisms on viral infection of the intestinal tract. In this Progress article, I review the recent advances that elucidate the mechanisms by which enteric viruses use commensal bacteria to enhance viral infectivity. These mechanisms segregate into two general categories: the direct facilitation of viral infection, including bacterial stabilization of viral particles and the facilitation of viral attachment to host target cells; and the indirect skewing of the antiviral immune response in a manner that promotes viral infection. Finally, I discuss the implications of these interactions for the development of vaccines and novel therapeutic approaches.

  20. A mass balance approach to the fate of viruses in a municipal wastewater treatment plant during summer and winter seasons.

    PubMed

    Ulbricht, Katharina; Selinka, Hans-Christoph; Wolter, Stefanie; Rosenwinkel, Karl-Heinz; Nogueira, Regina

    2014-01-01

    In contrast to previous discussion on general virus removal efficiency and identifying surrogates for human pathogenic viruses, this study focuses on virus retention within each step of a wastewater treatment plant (WWTP). Additionally, the influence of weather conditions on virus removal was addressed. To account for the virus retention, this study describes a mass balance of somatic coliphages (bacterial viruses) in a municipal WWTP, performed in the winter and summer seasons of 2011. In the winter season, the concentration of coliphages entering the WWTP was about 1 log lower than in summer. The mass balance in winter revealed a virus inactivation of 85.12 ± 13.97%. During the summer season, virus inactivation was significantly higher (95.25 ± 3.69%, p-value <0.05), most likely due to additional virus removal in the secondary clarifier by insolation. Thus, a total removal of coliphages of about 2.78 log units was obtained in summer compared to 1.95 log units in winter. Rainfall events did not statistically correlate with the concentrations of coliphages entering the WWTP in summer.

  1. Heat inactivation of enteric viruses in dewatered wastewater sludge.

    PubMed

    Ward, R L; Ashley, C S

    1978-12-01

    The effect of moisture content on the rates of heat inactivation of enteric viruses in wastewater sludge was determined. The protective effect of raw sludge on poliovirus previously observed (R. L. Ward, C. S. Ashley, and R. H. Moseley, Appl. Environ. Microbiol. 32:339--346, 1976) was found to be greatly enhanced in sludge dewatered by evaporation. Other enteroviruses responded in a similar fashion. This effect did not appear to be due merely to the state of dryness of the sludge samples because in humus-deficient soil, a relatively inert material, the rate of poliovirus inactivation by heat was not significantly altered through dewatering. Instead, this effect appeared to have been caused by protective substances in the sludge, such as detergents, which are concentrated through dewatering. As reported previously (R. L. Ward and C. S. Ashley, Appl. Environ. Microbiol. 34:681-688, 1977; R. L. Ward and C. S. Ashley, Appl. Environ. Microbiol 36:889--897, 1978) raw sludge is not protective of reovirus, but, instead, the ionic detergents in sludge cause the rate of heat inactivation of this virus to be accelerated. Dewatering of sludge, however, was found to partially reverse this virucidal effect. Evidence is presented indicating that this reversal is caused by an unidentified protective substance in sludge also concentrated through dewatering. Finally, it was shown that the effects of raw sludge on heat inactivation of poliovirus and reovirus are greatly reduced by composting, a result that correlated with the degradation of detergents.

  2. Dynamics of Virus Distribution in a Defined Swine Production Network Using Enteric Viruses as Molecular Markers.

    PubMed

    Lachapelle, Virginie; Letellier, Ann; Fravalo, Philippe; Brassard, Julie; L'Homme, Yvan

    2017-02-15

    Modern swine production systems represent complex and dynamic networks involving numerous stakeholders. For instance, livestock transporters carry live animals between fattening sites, abattoirs, and other premises on a daily basis. This interconnected system may increase the risk of microbial spread within and between networks, although little information is available in that regard. In the present study, a swine network composed of 10 finishing farms, one abattoir, and three types of stakeholders (veterinarians, livestock transporters, and nutritional technicians) in Quebec, Canada, was selected to investigate specific vectors and reservoirs of enteric viruses. Environmental samples were collected from the premises over a 12-month period. Samples were screened using targeted reverse transcription-PCR and sequencing of two selected viral markers, group A rotaviruses (RVA) and porcine astroviruses (PoAstV), both prevalent and genetically heterogeneous swine enteric viruses. The results revealed frequent contamination of farm sites (21.4 to 100%), livestock transporter vehicles (30.6 to 68.8%) and, most importantly, the abattoir yard (46.7 to 94.1%), depending on the sample types. Although high levels of strain diversity for both viruses were found, identical PoAstV and RVA strains were detected in specific samples from farms, the abattoir yard, and the livestock transporter vehicle, suggesting interconnections between these premises and transporters. Overall, the results from this study underscore the potential role of abattoirs and livestock transport as a reservoir and transmission route for enteric viruses within and between animal production networks, respectively.

  3. Differential depuration of poliovirus, Escherichia coli, and a coliphage by the common mussel, Mytilus edulis

    SciTech Connect

    Power, U.F.; Collins, J.K. )

    1989-06-01

    The elimination of sewage effluent-associated poliovirus, Escherichia coli, and a 22-nm icosahedral coliphage by the common mussel, Mytilus edulis, was studied. Both laboratory-and commercial-scale recirculating, UV depuration systems were used in this study. In the laboratory system, the logarithms of the poliovirus, E. coli, and coliphage levels were reduced by 1.86, 2.9, and 2.16, respectively, within 52 h of depuration. The relative patterns and rates of elimination of the three organisms suggest that they are eliminated from mussels by different mechanisms during depuration under suitable conditions. Poliovirus was not included in experiments undertaken in the commercial-scale depuration system. The differences in the relative rates and patterns of elimination were maintained for E. coli and coliphage in this system, with the logarithm of the E. coli levels being reduced by 3.18 and the logarithm of the coliphage levels being reduced by 0.87. The results from both depuration systems suggest that E. coli is an inappropriate indicator of the efficiency of virus elimination during depuration. The coliphage used appears to be a more representative indicator. Depuration under stressful conditions appeared to have a negligible affect on poliovirus and coliphage elimination rates from mussels. However, the rate and pattern of E. coli elimination were dramatically affected by these conditions. Therefore, monitoring E. coli counts might prove useful in ensuring that mussels are functioning well during depuration.

  4. [Morphological characteristics of haemorrhagic enteritis in dogs caused by parvo-like viruses (author's transl)].

    PubMed

    v d Gaag, I; van den Ingh, T S; van Dijk, J E

    1980-03-15

    Various outbreaks of parvo-like virus infection in dogs are reported. A form of haemorrhagic enteritis was observed, which was microscopically characterized by a hypo-regenerative villous atrophy of the small intestine, which bears a close resemblance to the typical lesion of feline panleucopenia. This pathomorphological feature may be regarded as typical of canine enteritis due to a parvo-like virus.

  5. Retention of Enteric Viruses by the Hemocytes of the Eastern Oyster (Crassostrea virginica)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Shellfish are an important vector for transmission of enteric pathogens. Interventions, such as depuration, do not adequately clear enteric viruses, while fecal bacteria levels are significantly reduced. Why viruses are retained in the bivalve flesh is not well understood. We hypothesize that phagoc...

  6. IMPROVED DETECTION OF HUMAN ENTERIC VIRUSES IN FOODS BY RT-PCR. (R826139)

    EPA Science Inventory

    Human enteric viruses (including hepatitis A virus (HAV) and Norwalk-like viruses (NLVs)) are now recognized as common causes of foodborne disease. While methods to detect these agents in clinical specimens have improved significantly over the last 10 years, applications to fo...

  7. Source and transport of human enteric viruses in deep municipal water supply wells

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Until recently, few water utilities or researchers were aware of possible virus presence in deep aquifers and wells. Over the past several years, repeated detection of enteric viruses in water from deep wells in south-central Wisconsin, shows that viruses can be significant groundwater contaminants ...

  8. Hydrogeological and statistical evidence for wide-spread enteric virus contamination of deep municipal wells

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Over the past eight years our research group has repeatedly detected human enteric viruses in water produced from deep (over 800 ft) bedrock water-supply wells in Madison, WI. The likely source of the viruses is leakage from urban sewers. These virus detections have been surprising because human ent...

  9. DEVELOPMENT OF HOMOLOGOUS VIRAL INTERNAL CONTROLS FOR USE IN RT-PCR ASSAYS OF WATERBORNE ENTERIC VIRUSES

    EPA Science Inventory

    Enteric viruses often contaminate water sources causing frequent outbreaks of gastroenteritis. Reverse transcription-polymerase chain reaction (RT-PCR) assays are commonly used for detection of human enteric viruses in environmental and drinking water samples. RT-PCR provides ...

  10. Coliphage and indigenous phage in Mamala Bay, Oahu, Hawaii.

    PubMed

    Paul, J H; Rose, J B; Jiang, S C; London, P; Xhou, X; Kellogg, C

    1997-01-01

    Public concern over the discharge of primarily treated sewage by two offshore outfalls in Mamala Bay, Oahu, prompted a multidisciplinary study to determine the impact of such activities on the water quality in the bay and at adjacent recreational beaches. As part of this study, we determined the abundance of coliphage as an indicator of fecal pollution along with total viral direct counts and phages infective for Vibrio parahaemoltyicus 16 at stations in Mamala Bay in four quarterly samplings over 13 months. Coliphage (< 1 to 1.2 x 10(3)/liter) were found during each quarterly sampling along an offshore transect to the Sand Island waste treatment facility outfall. The nonpoint coastal stations (Pearl Harbor, Ala Wai Canal, and Ke'ehi Lagoon) had high levels of coliphage during the storm event sampling in February 1994 but much lower levels or none when sampled during dry weather. Coliphage were absent at all samplings at Waikiki Beach and at the control station off Diamond Head. Viral direct counts in eutrophic coastal stations (Pearl Harbor, Ke'ehi Lagoon, Ala Moana Beach, and Ala Wai canal) averaged 10(9)/liter, while counts at offshore stations ranged from 9 x 10(7) to 1 x 10(9) viruses/liter, values similar to those for other marine environments. Vibriophage were found mainly in eutrophic coastal environments (Ala Wai Canal, Pearl Harbor, and Ke'ehi Lagoon) and at the Sand Island Transect stations D1 and D2. The greatest abundance was found during the storm event (February 1994) sampling. These results suggest that the Sand Island outfall influenced the water quality of the immediate surrounding waters but had little effect on the quality of the recreational beaches. Nonpoint discharge sources appeared to be more important in the distribution of fecal indicators in the coastal zone.

  11. The effect of chlorine and combined chlorine/UV treatment on coliphages in drinking water disinfection.

    PubMed

    Zyara, Alyaa M; Torvinen, Eila; Veijalainen, Anna-Maria; Heinonen-Tanski, Helvi

    2016-08-01

    Chlorine disinfection is a globally used method to ensure the safety of drinking water. However, it has not always been successful against viruses and, therefore, it is important to find new methods to disinfect water. Seventeen different coliphages were isolated from the treated municipal wastewater. These coliphages and MS2 were treated with different dosages of chlorine in drinking water, and a combined chlorine/ultraviolet irradiation treatment for the chlorine-resistant coliphages. Chlorine disinfection with 0.3-0.5 mg/L total chlorine (free Cl-dosage 0.12-0.21 mg/L) for 10 min achieved 2.5-5.7 Log10-reductions for 11 sensitive coliphages. The six most resistant coliphages showed no reduction with these chlorine concentrations. MS2 was intermediate in chlorine resistance, and thus it is not a good indicator for viruses in chlorine disinfection. In the combined treatment total chlorine of 0.05-0.25 mg/L (free Cl-dosage 0.02-0.08 mg/L) and ultraviolet irradiation (14-22 mWs/cm(2)) were more effective than chlorine alone, and 3-5 Log10-reductions were achieved for the chlorine-resistant strains. The chlorination efficiency could be increased by higher dosages and longer contact times, but this could increase the formation of disinfection by-products. Therefore, the combination treatment is a recommended disinfection method.

  12. Pathogenic simian immunodeficiency virus infection is associated with expansion of the enteric virome.

    PubMed

    Handley, Scott A; Thackray, Larissa B; Zhao, Guoyan; Presti, Rachel; Miller, Andrew D; Droit, Lindsay; Abbink, Peter; Maxfield, Lori F; Kambal, Amal; Duan, Erning; Stanley, Kelly; Kramer, Joshua; Macri, Sheila C; Permar, Sallie R; Schmitz, Joern E; Mansfield, Keith; Brenchley, Jason M; Veazey, Ronald S; Stappenbeck, Thaddeus S; Wang, David; Barouch, Dan H; Virgin, Herbert W

    2012-10-12

    Pathogenic simian immunodeficiency virus (SIV) infection is associated with enteropathy, which likely contributes to AIDS progression. To identify candidate etiologies for AIDS enteropathy, we used next-generation sequencing to define the enteric virome during SIV infection in nonhuman primates. Pathogenic, but not nonpathogenic, SIV infection was associated with significant expansion of the enteric virome. We identified at least 32 previously undescribed enteric viruses during pathogenic SIV infection and confirmed their presence by using viral culture and PCR testing. We detected unsuspected mucosal adenovirus infection associated with enteritis as well as parvovirus viremia in animals with advanced AIDS, indicating the pathogenic potential of SIV-associated expansion of the enteric virome. No association between pathogenic SIV infection and the family-level taxonomy of enteric bacteria was detected. Thus, enteric viral infections may contribute to AIDS enteropathy and disease progression. These findings underline the importance of metagenomic analysis of the virome for understanding AIDS pathogenesis.

  13. Enteric Virus Survival during Household Laundering and Impact of Disinfection with Sodium Hypochlorite▿

    PubMed Central

    Gerba, Charles P.; Kennedy, Denise

    2007-01-01

    This study was conducted to determine whether enteric viruses (adenovirus, rotavirus, and hepatitis A virus) added to cotton cloth swatches survive the wash cycle, the rinse cycle, and a 28-min permanent press drying cycle as commonly practiced in households in the United States. Detergent with and without bleach (sodium hypochlorite) was added to washing machines containing sterile and virus-inoculated 58-cm2 swatches, 3.2 kg of cotton T-shirts and underwear, and a soiled pillowcase designed to simulate the conditions (pH, organic load, etc.) encountered in soiled laundry. The most important factors for the reduction of virus in laundry were passage through the drying cycle and the addition of sodium hypochlorite. Washing with detergent alone was not found to be effective for the removal or inactivation of enteric viruses, as significant concentrations of virus were found on the swatches (reductions of 92 to 99%). It was also demonstrated that viruses are readily transferred from contaminated cloths to uncontaminated clothes. The use of sodium hypochlorite reduced the number of infectious viruses on the swatches after washing and drying by at least 99.99%. Laundering practices in common use in the United States do not eliminate enteric and respiratory viruses from clothes. The use of bleach can further reduce the numbers of enteric viruses in laundry. PMID:17526793

  14. Tissue distribution of a coliphage and Escherichia coli in mussels after contamination and depuration.

    PubMed Central

    Power, U F; Collins, J K

    1990-01-01

    Experiments were undertaken to determine the tissue distribution of Escherichia coli and a coliphage after contamination of the common mussel (Mytilus edulis). Mussels were contaminated with high levels of feces-associated E. coli and a 22-nm icosahedral coliphage over a 2-day period in a flowing-seawater facility. After contamination, individual tissues were carefully dissected and assayed for E. coli and the coliphage. Contaminated mussels were also analyzed to determine the tissue distribution of the contaminants after 24- and 48-h depuration periods. The majority of each contaminant was located in the digestive tract (94 and 89% of E. coli and coliphage, respectively). Decreasing concentrations were found in the gills and labial palps, foot and muscles, mantle lobes, and hemolymph. Our results indicate that contamination above levels in water occurred only in the digestive tract. Contaminated mussels were depurated in a commercial-scale recirculating UV depuration system over a 48-h period. The percent reductions of E. coli occurred in the following order: digestive tract, hemolymph, foot and muscles, mantle lobes, and gills and labial palps. The percent reductions of the coliphage were different, occurring in the following order: hemolymph, foot and muscles, gills and labial palps, mantle lobes, and digestive tract. Our results clearly demonstrate that E. coli and the coliphage are differentially eliminated from the digestive tract. The two microorganisms are eliminated at similar rates from the remaining tissues. Our results also clearly show that the most significant coliphage retention after depuration for 48 h is in the digestive tract. Thus, conventional depuration practices are inappropriate for efficient virus elimination from mussels. PMID:2180372

  15. Isolation and immunisation studies of a canine parco-like virus from dogs with haemorrhagic enteritis.

    PubMed

    Appel, M J; Scott, F W; Carmichael, L E

    1979-08-25

    A newly recognised canine parvo like virus was isolated from faeces of dogs with haemorrhagic enteritis. Cell cultures from several species were susceptible to it. Virus infected cells could be demonstrated by staining with fluorescent antibody reagents (prepared against canine virus or feline panleucopenia virus) or by haemagglutination with pig or rhesus monkey red blood cells. Inhibition of haemagglutination by specific antiserum prepared in specific-pathogen-free beagles provided a convenient method for viral identification. Experimental inoculation of specific-pathogen-free beagles resulted in elevated body temperatures and caused lymphopenia lasting one to three days. Feline panleucopenia virus vaccines protected dogs against challenge with virulent canine parvo-like virus.

  16. Application of enteric viruses for fecal pollution source tracking in environmental waters

    EPA Science Inventory

    Microbial source tracking (MST) tools are used to identify sources of fecal pollution for accurately assessing public health risk and implementing best management practices (BMPs). This review focuses on the potential of enteric viruses for MST applications. Following host infect...

  17. Groundwater sampling methods using glass wool filtration to trace human enteric viruses in Madison, Wisconsin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Human enteric viruses have been detected in the Madison, Wisconsin deep municipal well system. Earlier projects by the Wisconsin Geological and Natural History Survey (WGNHS) have used glass wool filters to sample groundwater for these viruses directly from the deep municipal wells. Polymerase chain...

  18. Retention of Enteric Viruses by the Hemocytes of the Eastern Oyster (Crassostrea virginica)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Virus accumulation and persistence in bivalve mollusks has long been documented in the United States and also throughout the world. Shellfish are an important vector for transmission of enteric pathogens. Interventions, such as depuration, do not adequately clear oysters of virus, while fecal bacter...

  19. Human enteric viruses in groundwater from a confined bedrock aquifer

    USGS Publications Warehouse

    Borchardt, M. A.; Bradbury, K.R.; Gotkowitz, M.B.; Cherry, J.A.; Parker, B.L.

    2007-01-01

    Confined aquifers are overlain by low-permeability aquitards that are commonly assumed to protect underlying aquifers from microbial contaminants. However, empirical data on microbial contamination beneath aquitards is limited. This study determined the occurrence of human pathogenic viruses in well water from a deep sandstone aquifer confined by a regionally extensive shale aquitard. Three public water-supply wells were each sampled 10 times over 15 months. Samples were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) for several virus groups and by cell culture for infectious enteroviruses. Seven of 30 samples were positive by RT-PCR for enteroviruses; one of these was positive for infectious echovirus 18. The virus-positive samples were collected from two wells cased through the aquitard, indicating the viruses were present in the confined aquifer. Samples from the same wells showed atmospheric tritium, indicating water recharged within the past few decades. Hydrogeologic conditions support rapid porous media transport of viruses through the upper sandstone aquifer to the top of the aquitard 61 m below ground surface. Natural fractures in the shale aquitard are one possible virus transport pathway through the aquitard; however, windows, cross-connecting well bores, or imperfect grout seals along well casings also may be involved. Deep confined aquifers can be more vulnerable to contamination by human viruses than commonly believed. ?? 2007 American Chemical Society.

  20. Inadequately Treated Wastewater as a Source of Human Enteric Viruses in the Environment

    PubMed Central

    Okoh, Anthony I.; Sibanda, Thulani; Gusha, Siyabulela S.

    2010-01-01

    Human enteric viruses are causative agents in both developed and developing countries of many non-bacterial gastrointestinal tract infections, respiratory tract infections, conjunctivitis, hepatitis and other more serious infections with high morbidity and mortality in immunocompromised individuals such as meningitis, encephalitis and paralysis. Human enteric viruses infect and replicate in the gastrointestinal tract of their hosts and are released in large quantities in the stools of infected individuals. The discharge of inadequately treated sewage effluents is the most common source of enteric viral pathogens in aquatic environments. Due to the lack of correlation between the inactivation rates of bacterial indicators and viral pathogens, human adenoviruses have been proposed as a suitable index for the effective indication of viral contaminants in aquatic environments. This paper reviews the major genera of pathogenic human enteric viruses, their pathogenicity and epidemiology, as well as the role of wastewater effluents in their transmission. PMID:20644692

  1. Occurrence and distribution of microbiological contamination and enteric viruses in shallow ground water in Baltimore and Harford counties, Maryland

    USGS Publications Warehouse

    Banks, William S.L.; Battigelli, David A.

    2002-01-01

    The U.S. Geological Survey, in cooperation with the Maryland Department of the Environment and the Wisconsin State Laboratory of Hygiene, conducted a study to characterize the occurrence and distribution of viral contamination in small (withdrawing less than 10,000 gallons per day) public water-supply wells screened in the shallow aquifer in the Piedmont Physiographic Province in Baltimore and Harford Counties, Maryland. Two hundred sixty-three small public water-supply wells were in operation in these counties during the spring of 2000. Ninety-one of these sites were selected for sampling using a methodology that distributed the samples evenly over the population and the spatial extent of the study area. Each site, and its potential susceptibility to microbiological contamination, was evaluated with regard to hole depth, casing interval, and open interval. Each site was evaluated using characteristics such as on-site geology and on-site land use.Samples were collected by pumping between 200 and 400 gallons of untreated well water through an electropositive cartridge filter. Water concentrates were subjected to cell-culture assay for the detection of culturable viruses and reverse-transcription polymerase chain reaction/gene probe assays to detect viral ribonucleic acid; grab samples were analyzed for somatic and male-specific coliphages, Bacteroides fragilis, Clostridium perfringens, enterococci, Escherichia coli, total coliforms, total oxidized nitrogen, nitrite, organic nitrogen, total phosphate, ortho-phosphate, calcium, magnesium, sodium, potas-sium, chloride, sulfate, iron, acid-neutralizing capacity, pH, specific conductance, temperature, and dissolved oxygen.One sample tested positive for the presence of the ribonucleic acid of rotavirus through poly-merase chain-reaction analysis. Twenty-nine per-cent of the samples (26 of 90) had bacterial con-tamination. About 7 percent of the samples (6 of 90) were contaminated with either male-specific coliphage

  2. Efficient strategy for constructing duck enteritis virus-based live attenuated vaccine against homologous and heterologous H5N1 avian influenza virus and duck enteritis virus infection.

    PubMed

    Zou, Zhong; Hu, Yong; Liu, Zhigang; Zhong, Wei; Cao, Hangzhou; Chen, Huanchun; Jin, Meilin

    2015-04-16

    Duck is susceptible to many pathogens, such as duck hepatitis virus, duck enteritis virus (DEV), duck tembusu virus, H5N1 highly pathogenic avian influenza virus (HPAIV) in particular. With the significant role of duck in the evolution of H5N1 HPAIV, control and eradication of H5N1 HPAIV in duck through vaccine immunization is considered an effective method in minimizing the threat of a pandemic outbreak. Consequently, a practical strategy to construct a vaccine against these pathogens should be determined. In this study, the DEV was examined as a candidate vaccine vector to deliver the hemagglutinin (HA) gene of H5N1, and its potential as a polyvalent vaccine was evaluated. A modified mini-F vector was inserted into the gB and UL26 gene junction of the attenuated DEV vaccine strain C-KCE genome to generate an infectious bacterial artificial chromosome (BAC) of C-KCE (vBAC-C-KCE). The HA gene of A/duck/Hubei/xn/2007 (H5N1) was inserted into the C-KCE genome via the mating-assisted genetically integrated cloning (MAGIC) to generate the recombinant vector pBAC-C-KCE-HA. A bivalent vaccine C-KCE-HA was developed by eliminating the BAC backbone. Ducks immunized with C-KCE-HA induced both the cross-reactive antibodies and T cell response against H5. Moreover, C-KCE-HA-immunized ducks provided rapid and long-lasting protection against homologous and heterologous HPAIV H5N1 and DEV clinical signs, death, and primary viral replication. In conclusion, our BAC-C-KCE is a promising platform for developing a polyvalent live attenuated vaccine.

  3. Adhesion of human pathogenic enteric viruses and surrogate viruses to inert and vegetal food surfaces.

    PubMed

    Deboosere, Nathalie; Pinon, Anthony; Caudrelier, Yvette; Delobel, Alexandre; Merle, Ghislaine; Perelle, Sylvie; Temmam, Sarah; Loutreul, Julie; Morin, Thierry; Estienney, Marie; Belliot, Gael; Pothier, Pierre; Gantzer, Christophe; Vialette, Michèle

    2012-10-01

    Enteric viruses, particularly human Noroviruses (NoV) and hepatitis A virus (HAV), are key food-borne pathogens. The attachment of these pathogens to foodstuff and food-contact surfaces is an important mechanism in the human contamination process. Studies were done to investigate the nature of the physicochemical forces, such as hydrophobic and electrostatic ones, involved in the interaction virus/matrix but, at this day, only few data are available concerning surface properties of viruses and prediction of the adhesion capacity of one specific virus onto matrices is still very difficult. The purpose of this study was to propose a reference system, including a representative virus surrogate, able to predict as close as possible behaviour of pathogenic viruses in term of adhesion on inert (stainless steel and polypropylene) and food surfaces (lettuce leaves, strawberries and raspberries). The adhesion of human pathogenic enteric viruses, cultivable strain of HAV and non-cultivable strains of human NoV (genogroups I and II), have been quantified and compared to these of human enteric viruses surrogates, included the MNV-1 and three F-specific RNA bacteriophages (MS2, GA and Qβ). A standardized approach was developed to assess and quantify viral adhesion on tested matrices after a contact time with each virus using real-time RT-PCR. Methods used for virus recovery were in accordance with the CEN recommendations, including a bovine Enterovirus type 1 as control to monitor the efficiency of the extraction process and amplification procedure from directly extracted or eluted samples. The adhesion of human pathogenic viruses, ranging from 0.1 to 2%, could be comparable for all matrices studied, except for NoV GII on soft fruits. Adhesion percentages obtained for the studied surrogate virus and phages were shown to be comparable to those of HAV and NoV on inert and lettuce surfaces. The MNV-1 appeared as the best candidate to simulate adhesion phenomena of all human

  4. Relationships Between Environmental Factors, Bacterial Indicators, and the Occurrence of Enteric Viruses in Estuarine Sediments

    PubMed Central

    LaBelle, Raymond L.; Gerba, Charles P.; Goyal, Sagar M.; Melnick, Joseph L.; Cech, Irina; Bogdan, Gregory F.

    1980-01-01

    Current standards for evaluation of the public health safety of recreational and shellfish-harvesting waters are based upon bacteriological analysis, but do not include an evaluation of the number of viruses. The objective of this study was to determine the occurrence of enteric viruses in estuarine sediments and to find a relationship, if any, between the presence of viruses in seawater or sediment or both and various biological and physicochemical characteristics of the environment. Viruses were found in greater numbers in sediment than in overlying seawater on a volume basis. Several types of enteroviruses were isolated: coxsackievirus types A16, B1, and B5, echovirus type 1, and poliovirus type 2. On several occasions, viruses were isolated from sediments when overlying seawaters met bacteriological water quality standards for recreational use. Statistical analysis of the relationship between viruses in seawater or in sediment and other variables measured yielded only one significant association: the number of viruses in sediment was found to be positively correlated with the number of fecal coliforms in sediment. No other physical, chemical, or biological characteristic of seawater or sediment that was measured showed statistically significant association with viral numbers. No correlation was found between bacterial indicators and virus in the overlying waters. The data indicated that evaluation of the presence of bacteria and viruses in sediment may provide additional insight into long-term water quality conditions and that indicator bacteria in water are not reflective of the concentration of enteric viruses in marine waters. PMID:6247974

  5. Evaluation of Human Enteric Viruses in Surface Water and Drinking Water Resources in Southern Ghana

    PubMed Central

    Gibson, Kristen E.; Opryszko, Melissa C.; Schissler, James T.; Guo, Yayi; Schwab, Kellogg J.

    2011-01-01

    An estimated 884 million people worldwide do not have access to an improved drinking water source, and the microbial quality of these sources is often unknown. In this study, a combined tangential flow, hollow fiber ultrafiltration (UF), and real-time PCR method was applied to large volume (100 L) groundwater (N = 4), surface water (N = 9), and finished (i.e., receiving treatment) drinking water (N = 6) samples for the evaluation of human enteric viruses and bacterial indicators. Human enteric viruses including norovirus GI and GII, adenovirus, and polyomavirus were detected in five different samples including one groundwater, three surface water, and one drinking water sample. Total coliforms and Escherichia coli assessed for each sample before and after UF revealed a lack of correlation between bacterial indicators and the presence of human enteric viruses. PMID:21212196

  6. Evaluation of human enteric viruses in surface water and drinking water resources in southern Ghana.

    PubMed

    Gibson, Kristen E; Opryszko, Melissa C; Schissler, James T; Guo, Yayi; Schwab, Kellogg J

    2011-01-01

    An estimated 884 million people worldwide do not have access to an improved drinking water source, and the microbial quality of these sources is often unknown. In this study, a combined tangential flow, hollow fiber ultrafiltration (UF), and real-time PCR method was applied to large volume (100 L) groundwater (N = 4), surface water (N = 9), and finished (i.e., receiving treatment) drinking water (N = 6) samples for the evaluation of human enteric viruses and bacterial indicators. Human enteric viruses including norovirus GI and GII, adenovirus, and polyomavirus were detected in five different samples including one groundwater, three surface water, and one drinking water sample. Total coliforms and Escherichia coli assessed for each sample before and after UF revealed a lack of correlation between bacterial indicators and the presence of human enteric viruses.

  7. Metagenomic analysis of the shrew enteric virome reveals novel viruses related to human stool-associated viruses.

    PubMed

    Sasaki, Michihito; Orba, Yasuko; Ueno, Keisuke; Ishii, Akihiro; Moonga, Ladslav; Hang'ombe, Bernard M; Mweene, Aaron S; Ito, Kimihito; Sawa, Hirofumi

    2015-02-01

    Shrews are small insectivorous mammals that are distributed worldwide. Similar to rodents, shrews live on the ground and are commonly found near human residences. In this study, we investigated the enteric virome of wild shrews in the genus Crocidura using a sequence-independent viral metagenomics approach. A large portion of the shrew enteric virome was composed of insect viruses, whilst novel viruses including cyclovirus, picornavirus and picorna-like virus were also identified. Several cycloviruses, including variants of human cycloviruses detected in cerebrospinal fluid and stools, were detected in wild shrews at a high prevalence rate. The identified picornavirus was distantly related to human parechovirus, inferring the presence of a new genus in this family. The identified picorna-like viruses were characterized as different species of calhevirus 1, which was discovered previously in human stools. Complete or nearly complete genome sequences of these novel viruses were determined in this study and then were subjected to further genetic characterization. Our study provides an initial view of the diversity and distinctiveness of the shrew enteric virome and highlights unique novel viruses related to human stool-associated viruses.

  8. Detection of enteric viruses in activated sludge by feasible concentration methods.

    PubMed

    Prado, Tatiana; Gaspar, Ana Maria Coimbra; Miagostovich, Marize Pereira

    2014-01-01

    Human enteric viruses are responsible to cause several diseases, including gastroenteritis and hepatitis, and can be present in high amounts in sewage sludge. This study compared virus recovery efficiency of two feasible concentration methods used for detecting human adenovirus (HAdV), rotavirus species A (RV-A), norovirus genogroup II (NoV GII) and hepatitis A virus (HAV) in sewage sludge from an activated sludge process. Twelve sewage sludge samples were collected bi-monthly from January to July, 2011. Ultracentrifugation was compared with a simplified protocol based on beef extract elution for recovering enteric viruses. Viruses were quantified by quantitative real-time PCR assays and virus recovery efficiency and limits of detection were determined. Methods showed mean recovery rates lower than 7.5%, presenting critical limits of detection (higher than 10(2) - 10(3) genome copies - GC L(-1) for all viruses analyzed). Nevertheless, HAdV were detected in 90% of the analyzed sewage sludge samples (range: 1.8 × 10(4) to 1.1 × 10(5) GC L(-1)), followed by RV-A and NoV (both in 50%) and HAV (8%). Results suggesting that activated sludge is contaminated with high viral loads and HAdV are widely disseminated in these samples. The low virus recovery rates achieved, especially for HAV, indicate that other feasible concentration methods could be developed to improve virus recovery efficiency in these environmental matrices.

  9. Techniques for the recovery of enteric viruses from artificially contaminated marine sediments.

    PubMed

    Pianetti, Anna; Citterio, Barbara; Sabatini, Luigia; Pierfelici, Lucia; Colantoni, Paolo; Bruscolini, Francesca

    2007-01-01

    Viruses are an important component of aquatic microbial communities and marine sediments may represent an optimal means for their survival. The aim of this study was to evaluate different methods for virus recovery from marine sediments. Three methods were used for virus recovery from artificially contaminated sediments: elution and centrifugation technique, sonication technique, and mechanical disgregation followed by elution and centrifuge technique. The sonication technique obtained the highest virus recovery percentages (94,25%). Eluent 2 provided more efficient recovery of enteric viruses than eluent 1 presumably due to the presence, in eluent 2, of NANO3, a chaotropic agent that enhances the solubilization of hydrophobic compounds in water. Finally, the authors confirm the importance of searching for viruses in sediments, which protect them from inactivation by biological, chemical and physical factors and allow them to survive longer than in the overlaying water column.

  10. Case report: epithelial intracytoplasmic herpes viral inclusions associated with an outbreak of duck virus enteritis

    USGS Publications Warehouse

    Barr, B.C.; Jessup, David A.; Docherty, Douglas E.; Lownestine, L.J.

    1992-01-01

    Several muscovy ducks from a free-roaming flock of 65 muscovy and mallard ducks died over a 3-week period. Three muscovy ducks were necropsied. Gross and microscopic changes were compatible with duck virus enteritis, and the virus was isolated. In addition to intranuclear viral inclusion bodies in several tissues, intracytoplasmic inclusion bodies were present in esophageal and cloacal epithelium, By electron microscopy, the membrane-bound intracytoplasmic inclusions were found to contain enveloped herpesvirus, and nuclei contained herpes viral nucleocapsids.

  11. Thermal Inactivation of Foodborne Enteric Viruses and Their Viral Surrogates in Foods.

    PubMed

    Bozkurt, Hayriye; D'Souza, Doris H; Davidson, P Michael

    2015-08-01

    Foodborne viruses, in particular human norovirus and hepatitis A virus, are the most common causes of food-associated infections and foodborne illness outbreaks around the world. Since it is currently not possible to cultivate human noroviruses and the wild-type strain of hepatitis A virus in vitro, the use of a variety of viral surrogates is essential to determine appropriate thermal processing conditions to reduce the risk associated with their contamination of food. Therefore, the objectives of this review are to (i) present pertinent characteristics of enteric foodborne viruses and their viral surrogates, (ii) discuss the viral surrogates currently used in thermal inactivation studies and their significance and value, (iii) summarize available data on thermal inactivation kinetics of enteric viruses, (iv) discuss factors affecting the efficacy of thermal treatment, (v) discuss suggested mechanisms of thermal inactivation, and (vi) provide insights on foodborne enteric viruses and viral surrogates for future studies and industrial applications. The overall goal of this review is to contribute to the development of appropriate thermal processing protocols to ensure safe food for human consumption.

  12. Presence of enteric viruses in freshwater and their removal by the conventional drinking water treatment process.

    PubMed Central

    Hurst, C. J.

    1991-01-01

    A review of results published in English or French between 1980 and 1990 was carried out to determine the levels of indigenous human enteric viruses in untreated surface and subsurface freshwaters, as well as in drinking water that had undergone the complete conventional treatment process. For this purpose, the conventional treatment process was defined as an operation that included coagulation followed by sedimentation, filtration, and disinfection. Also assessed was the stepwise efficiency of the conventional treatment process, as practised at full-scale facilities, for removing indigenous viruses from naturally occurring freshwaters. A list was compiled of statistical correlations relating to the occurrence of indigenous viruses in water. PMID:1647273

  13. PRESENCE OF ENTERIC VIRUSES IN FRESHWATER AND THEIR REMOVAL BY THE CONVENTIONAL DRINKING WATER TREATMENT PROCESS

    EPA Science Inventory

    A review of results published in English or French between 1980 and 1990 was carried out to determine the levels of indigenous human enteric viruses in untreated surface and subsurface freshwaters, as well as in drinking water that had undergone the complete conventional treatmen...

  14. A MULTIPLEX REVERSE TRANSCIPTION-PCR METHOD FOR DETECTION OF HUMAN ENTERIC VIRUSES IN GROUNDWATER

    EPA Science Inventory

    Untreated groundwater is responsible for about half of the waterborne disease outbreaks in the United States. Human enteric viruses are thought to be leading etiological agents of many of these outbreaks, but there is relatively little information on the types and levels of viru...

  15. Effects of sanitation, freezing and frozen storage on enteric viruses in berries and herbs.

    PubMed

    Butot, S; Putallaz, T; Sánchez, G

    2008-08-15

    Norovirus (NV) and hepatitis A virus (HAV) are foodborne enteric viruses associated with outbreaks of disease following consumption of fresh or frozen produce. Model experiments were performed to determine the effectiveness of certain commercial processes for the removal of enteric viruses that might be present in berries and herbs. The survival and persistence of HAV, NV, rotavirus (RV) and feline calicivirus (FCV), a surrogate for NV, in frozen produce over time were determined. Survival and inactivation of HAV, RV and FCV were assessed by viral culture and quantitative reverse transcription-PCR (RT-PCR), whereas NV persistence was determined by quantitative RT-PCR only. Freezing did not significantly reduce the viability of any of the viruses except the infectivity of FCV in strawberries. Frozen storage for 3 months had limited effects on HAV and RV survival in all tested food products, whereas in frozen raspberries and strawberries FCV infectivity showed the highest decay rate due to acid pH. To simulate postharvesting conditions, fresh berries and herbs were rinsed with tap, warm or chlorinated water or with a chlorine dioxide (ClO(2)) solution. Available chlorine at a concentration of 200 ppm and ClO(2) at 10 ppm reduced measurable enteric viruses in raspberry and parsley samples by less than 2 log(10) units.

  16. Diversity of somatic coliphages in coastal regions with different levels of anthropogenic activity in São Paulo State, Brazil.

    PubMed

    Burbano-Rosero, E M; Ueda-Ito, M; Kisielius, J J; Nagasse-Sugahara, T K; Almeida, B C; Souza, C P; Markman, C; Martins, G G; Albertini, L; Rivera, I N G

    2011-06-01

    Bacteriophages are the most abundant and genetically diverse viruses on Earth, with complex ecology in both quantitative and qualitative terms. Somatic coliphages (SC) have been reported to be good indicators of fecal pollution in seawater. This study focused on determining the concentration of SC and their diversity by electron microscopy of seawater, plankton, and bivalve samples collected at three coastal regions in São Paulo, Brazil. The SC counts varied from <1 to 3.4 × 10(3) PFU/100 ml in seawater (73 samples tested), from <1 to 4.7 × 10(2) PFU/g in plankton (46 samples tested), and from <1 to 2.2 × 10(1) PFU/g in bivalves (11 samples tested). In seawater samples, a relationship between the thermotolerant coliforms and Escherichia coli and SC was observed at the three regions (P = 0.0001) according to the anthropogenic activities present at each region. However, SC were found in plankton samples from three regions: Baixada Santista (17/20), Canal de São Sebastião (6/14), and Ubatuba (3/12). In seawater samples collected from Baixada Santista, four morphotypes were observed: A1 (4.5%), B1 (50%), C1 (36.4%), and D1 (9.1%). One coliphage, Siphoviridae type T1, had the longest tail: between 939 and 995 nm. In plankton samples, Siphoviridae (65.8%), Podoviridae (15.8%), Microviridae (15.8%), and Myoviridae (2.6%) were found. In bivalves, only the morphotype B1 was observed. These SC were associated with enteric hosts: enterobacteria, E. coli, Proteus, Salmonella, and Yersinia. Baixada Santista is an area containing a high level of fecal pollution compared to those in the Canal de São Sebastião and Ubatuba. This is the first report of coliphage diversity in seawater, plankton, and bivalve samples collected from São Paulo coastal regions. A better characterization of SC diversity in coastal environments will help with the management and evaluation of the microbiological risks for recreation, seafood cultivation, and consumption.

  17. Intra-laboratory validation of a concentration method adapted for the enumeration of infectious F-specific RNA coliphage, enterovirus, and hepatitis A virus from inoculated leaves of salad vegetables.

    PubMed

    Dubois, Eric; Hennechart, Catherine; Deboosère, Nathalie; Merle, Ghislaine; Legeay, Odile; Burger, Christian; Le Calvé, Marie; Lombard, Bertrand; Ferré, Virginie; Traoré, Ousmane

    2006-04-25

    Salad vegetables exposed to fecal contamination may cause outbreaks of hepatitis or gastro-enteritis if they are eaten raw. A procedure, based on elution with phosphate-buffered saline and concentration by filtration through membrane filters, was developed for the recovery of enteric viruses from salad leaves. The method was evaluated using lettuce leaves inoculated with hepatitis A virus (HAV), poliovirus, and MS2 bacteriophage. In addition, this method was validated by an intra-laboratory study using leaves of various salad vegetables inoculated with MS2 phage. The French standard NF V 03-110 was used to establish the general principle and the technical protocol of the validation procedure. Linear regression models describing the quantitative reactions were good fits to data in the whole range of viral concentrations tested, which was from about 1 to 4 log plaque-forming units (PFU) per 25 g of lettuce. The fractions of inoculated viruses recovered were estimated to be about 64% for HAV, 18% for poliovirus, and 29% for MS2. No significant effect of the food matrix was found using various types of salad vegetable (butter lettuce, iceberg lettuce, romaine lettuce, witloof chicory, curly endive, corn salad, rocket and watercress). Moreover, the variance of the results was constant for all levels of virus contamination within the experimental range. Intermediate reproducibility experiments were also performed to allow calculation of the uncertainty factor, which was found to be 0.58 log PFU/25 g. When used in association with phage enumeration, this validated procedure is rapid enough to be used for screening salad vegetables for evaluation of the efficacy of processes for control of pathogenic microorganisms on such foods.

  18. Assessment of sewer source contamination of drinking water wells using tracers and human enteric viruses

    USGS Publications Warehouse

    Hunt, R.J.; Borchardt, M. A.; Richards, K.D.; Spencer, S. K.

    2010-01-01

    This study investigated the source, transport, and occurrence of human enteric viruses in municipal well water, focusing on sanitary sewer sources. A total of 33 wells from 14 communities were sampled once for wastewater tracers and viruses. Wastewater tracers were detected in four of these wells, and five wells were virus- positive by qRT-PCR. These results, along with exclusion of wells with surface water sources, were used to select three wells for additional investigation. Viruses and wastewater tracers were found in the groundwater at all sites. Some wastewater tracers, such as ionic detergents, flame retardants, and cholesterol, were considered unambiguous evidence of wastewater. Sampling at any given time may not show concurrent virus and tracer presence; however, given sufficient sampling over time, a relation between wastewater tracers and virus occurrence was identified. Presence of infectious viruses at the wellhead demonstrates that high-capacity pumping induced sufficiently short travel times for the transport of infectious viruses. Therefore, drinking-water wells are vulnerable to contaminants that travel along fast groundwater flowpaths even if they contribute a small amount of virus-laden water to the well. These results suggest that vulnerability assessments require characterization of "low yield-fast transport" in addition to traditional "high yield-slow transport", pathways. ?? 2010 American Chemical Society.

  19. Assessment of sewer source contamination of drinking water wells using tracers and human enteric viruses.

    PubMed

    Hunt, Randall J; Borchardt, Mark A; Richards, Kevin D; Spencer, Susan K

    2010-10-15

    This study investigated the source, transport, and occurrence of human enteric viruses in municipal well water, focusing on sanitary sewer sources. A total of 33 wells from 14 communities were sampled once for wastewater tracers and viruses. Wastewater tracers were detected in four of these wells, and five wells were virus- positive by qRT-PCR. These results, along with exclusion of wells with surface water sources, were used to select three wells for additional investigation. Viruses and wastewater tracers were found in the groundwater at all sites. Some wastewater tracers, such as ionic detergents, flame retardants, and cholesterol, were considered unambiguous evidence of wastewater. Sampling at any given time may not show concurrent virus and tracer presence; however, given sufficient sampling over time, a relation between wastewater tracers and virus occurrence was identified. Presence of infectious viruses at the wellhead demonstrates that high-capacity pumping induced sufficiently short travel times for the transport of infectious viruses. Therefore, drinking-water wells are vulnerable to contaminants that travel along fast groundwater flowpaths even if they contribute a small amount of virus-laden water to the well. These results suggest that vulnerability assessments require characterization of "low yield-fast transport" in addition to traditional "high yield-slow transport", pathways.

  20. AN UNEXPECTED TEMPORAL PATTERN OF COLIPHAGE ISOLATION IN GROUNDWATERS SAMPLED FROM WELLS AT VARIED DISTANCES FROM RECLAIMED WATER RECHARGE SITES

    EPA Science Inventory

    Potable and monitoring wells located in close proximity to a large groundwater recharge project which utilizes a blend of surface water and reclaimed wastewater for recharge were tested for coliphage over a period of 6 months to assess the potential for virus migration. During th...

  1. Simultaneous investigation of influenza and enteric viruses in the stools of adult patients consulting in general practice for acute diarrhea

    PubMed Central

    2012-01-01

    Background Gastrointestinal symptoms are not an uncommon manifestation of an influenza virus infection. In the present study, we aimed to investigate the presence of influenza viruses in the stools of adult patients consulting their general practitioner for uncomplicated acute diarrhea (AD) and the proportion of concurrent infections by enteric and influenza viruses. Method A case-control study was conducted from December 2010 to April 2011. Stool specimens were collected and tested for influenza viruses A (seasonal A/H3N2 and pandemic A/H1N1) and B, and for four enteric viruses (astrovirus, group A rotavirus, human enteric adenovirus, norovirus of genogroups I – NoVGI - and genogroup II - NoVGII). Results General practitioners enrolled 138 cases and 93 controls. Of the 138 stool specimens collected, 92 (66.7%) were positive for at least one of the four enteric viruses analysed and 10 (7.2%) tested positive for one influenza virus. None of these 10 influenza positive patients reported respiratory symptoms. In five influenza-positive patients (3.6%), we also detected one enteric virus, with 4 of them being positive for influenza B (2 had co-detection with NoVGI, 1 with NoVGII, and 1 with astrovirus). None of the 93 controls tested positive for one of the enteric and/or other influenza viruses we investigated. Conclusions In this study we showed that the simultaneous detection of influenza and enteric viruses is not a rare event. We have also reported, for the first time in general practice, the presence of seasonal and pandemic influenza viruses in the stools of adult patients consulting for uncomplicated AD. A simultaneous investigation of enteric and influenza viruses in patients complaining of gastrointestinal symptoms could be useful for future studies to better identify the agents responsible for AD. PMID:22709374

  2. Bacterial lipopolysaccharide binding enhances virion stability and promotes environmental fitness of an enteric virus.

    PubMed

    Robinson, Christopher M; Jesudhasan, Palmy R; Pfeiffer, Julie K

    2014-01-15

    Enteric viruses, including poliovirus and reovirus, encounter a vast microbial community in the mammalian gastrointestinal tract, which has been shown to promote virus replication and pathogenesis. Investigating the underlying mechanisms, we find that poliovirus binds bacterial surface polysaccharides, which enhances virion stability and cell attachment by increasing binding to the viral receptor. Additionally, we identified a poliovirus mutant, VP1-T99K, with reduced lipopolysaccharide (LPS) binding. Although T99K and WT poliovirus cell attachment, replication, and pathogenesis in mice are equivalent, VP1-T99K poliovirus was unstable in feces following peroral inoculation of mice. Consequently, the ratio of mutant virus in feces is reduced following additional cycles of infection in mice. Thus, the mutant virus incurs a fitness cost when environmental stability is a factor. These data suggest that poliovirus binds bacterial surface polysaccharides, enhancing cell attachment and environmental stability, potentially promoting transmission to a new host.

  3. An outbreak of duck virus enteritis (duck plague) in a captive flock of mixed waterfowl

    USGS Publications Warehouse

    Montgomery, R.D.; Stein, G.; Novilla, M.N.; Hurley, Sarah S.; Fink, R.J.

    1981-01-01

    An outbreak of duck virus enteritis occurred in a flock of captive waterfowl composed of mallards (Anas platyrhynchos), black ducks (Anas rubripes), and Canada geese (Branta canadensis). Although all three species were housed together, morbidity and mortality were confined to the 227 black ducks and Canada geese, of which 180 died and the rest were left in a weakened condition. Lesions are given for 20 black ducks and 4 Canada geese dying from DVE. In addition, both horizontal and vertical transmission are discussed as possible sources of the virus that caused this outbreak.

  4. Prevalence and Genetic Diversity of Enteric Viruses in Children with Diarrhea in Ouagadougou, Burkina Faso

    PubMed Central

    Ouédraogo, Nafissatou; Kaplon, Jérôme; Bonkoungou, Isidore Juste O.; Traoré, Alfred Sababénédjo; Pothier, Pierre; Barro, Nicolas; Ambert- Balay, Katia

    2016-01-01

    Enteric viruses are a major cause of diarrhea in children, especially those under five years old. Identifying the viral agents is critical to the development of effective preventive measures. This study aimed to determine the prevalence and genetic diversity of common enteric viruses in children under five years old in Burkina Faso. Stool samples from children with (n = 263) and without (n = 50) diarrhea disorders were collected in Ouagadougou, Burkina Faso from November 2011 to September 2012. Rotavirus, norovirus, sapovirus, astrovirus, adenovirus and Aichivirus A were detected using real-time or end-point (RT-)PCR. Rotavirus strains were G and P genotyped by multiplex RT-PCR and other viral strains were characterized by sequencing of viral subgenomic segements. At least one viral agent was detected in 85.6% and 72% of the symptomatic and asymptomatic patients, respectively. Rotavirus (63.5%), adenovirus (31.2%) and genogroup II norovirus (18.2%) were the most prevalent viruses in symptomatic patients, but only rotavirus and genogroup II norovirus were significantly associated with diarrhea (OR: 7.9, 95%CI: 3.7–17; OR: 3.5, 95%CI: 1–11.7, respectively). Sapovirus (10.3%), astrovirus (4.9%), genogroup I norovirus (2.7%) and Aichivirus A (0.8%) were less prevalent. The predominant genotype of rotavirus was G9P[8] (36.5%), and the predominant norovirus strain was GII.4 variant 2012 (71.4%). Among sapovirus, the genogroup II (87.5%) predominated. Astrovirus type 1 (41.7%) was the most frequent astrovirus identified. Aichivirus A belonged to the three genotypes (A, B and C). Enteric adenoviruses type 40 and 41 were identified in 10.2% and 5.1% respectively. Several cases of co-infections were detected. The results highlight the high prevalence and the high diversity of enteric viruses in Burkinabe children. PMID:27092779

  5. Prevalence and Genetic Diversity of Enteric Viruses in Children with Diarrhea in Ouagadougou, Burkina Faso.

    PubMed

    Ouédraogo, Nafissatou; Kaplon, Jérôme; Bonkoungou, Isidore Juste O; Traoré, Alfred Sababénédjo; Pothier, Pierre; Barro, Nicolas; Ambert-Balay, Katia

    2016-01-01

    Enteric viruses are a major cause of diarrhea in children, especially those under five years old. Identifying the viral agents is critical to the development of effective preventive measures. This study aimed to determine the prevalence and genetic diversity of common enteric viruses in children under five years old in Burkina Faso. Stool samples from children with (n = 263) and without (n = 50) diarrhea disorders were collected in Ouagadougou, Burkina Faso from November 2011 to September 2012. Rotavirus, norovirus, sapovirus, astrovirus, adenovirus and Aichivirus A were detected using real-time or end-point (RT-)PCR. Rotavirus strains were G and P genotyped by multiplex RT-PCR and other viral strains were characterized by sequencing of viral subgenomic segements. At least one viral agent was detected in 85.6% and 72% of the symptomatic and asymptomatic patients, respectively. Rotavirus (63.5%), adenovirus (31.2%) and genogroup II norovirus (18.2%) were the most prevalent viruses in symptomatic patients, but only rotavirus and genogroup II norovirus were significantly associated with diarrhea (OR: 7.9, 95%CI: 3.7-17; OR: 3.5, 95%CI: 1-11.7, respectively). Sapovirus (10.3%), astrovirus (4.9%), genogroup I norovirus (2.7%) and Aichivirus A (0.8%) were less prevalent. The predominant genotype of rotavirus was G9P[8] (36.5%), and the predominant norovirus strain was GII.4 variant 2012 (71.4%). Among sapovirus, the genogroup II (87.5%) predominated. Astrovirus type 1 (41.7%) was the most frequent astrovirus identified. Aichivirus A belonged to the three genotypes (A, B and C). Enteric adenoviruses type 40 and 41 were identified in 10.2% and 5.1% respectively. Several cases of co-infections were detected. The results highlight the high prevalence and the high diversity of enteric viruses in Burkinabe children.

  6. Assessment of the efficacy of membrane filtration processes to remove human enteric viruses and the suitability of bacteriophages and a plant virus as surrogates for those viruses.

    PubMed

    Shirasaki, N; Matsushita, T; Matsui, Y; Murai, K

    2017-05-15

    Here, we evaluated the efficacy of direct microfiltration (MF) and ultrafiltration (UF) to remove three representative human enteric viruses (i.e., adenovirus [AdV] type 40, coxsackievirus [CV] B5, and hepatitis A virus [HAV] IB), and one surrogate of human caliciviruses (i.e., murine norovirus [MNV] type 1). Eight different MF membranes and three different UF membranes were used. We also examined the ability of coagulation pretreatment with high-basicity polyaluminum chloride (PACl) to enhance virus removal by MF. The removal ratios of two bacteriophages (MS2 and φX174) and a plant virus (pepper mild mottle virus; PMMoV) were compared with the removal ratios of the human enteric viruses to assess the suitability of these viruses to be used as surrogates for human enteric viruses. The virus removal ratios obtained with direct MF with membranes with nominal pore sizes of 0.1-0.22 μm differed, depending on the membrane used; adsorptive interactions, particularly hydrophobic interactions between virus particles and the membrane surface, were dominant factors for virus removal. In contrast, direct UF with membranes with nominal molecular weight cutoffs of 1-100 kDa effectively removed viruses through size exclusion, and >4-log10 removal was achieved when a membrane with a nominal molecular weight cutoff of 1 kDa was used. At pH 7 and 8, in-line coagulation-MF with nonsulfated high-basicity PACls containing Al30 species had generally a better virus removal (i.e., >4-log10 virus removal) than the other aluminum-based coagulants, except for φX174. For all of the filtration processes, the removal ratios of AdV, CV, HAV, and MNV were comparable and strongly correlated with each other. The removal ratios of MS2 and PMMoV were comparable or smaller than those of the three human enteric viruses and MNV, and were strongly correlated with those of the three human enteric viruses and MNV. The removal ratios obtained with coagulation-MF for φX174 were markedly smaller than

  7. New strain of mouse hepatitis virus as the cause of lethal enteritis in infant mice.

    PubMed Central

    Hierholzer, J C; Broderson, J R; Murphy, F A

    1979-01-01

    A new strain of mouse hepatitis virus (MHV) was isolated from pooled gut suspensions from an epizootic of lethal enteritis in newborn mice. Negative-contrast electron microscopy showed an abundance of coronavirus particles in the intestinal contents and intestinal epithelium of moribund mice. We found no other virus in the epizootic. Dams seroconverted to MHV polyvalent antigen and to the agent isolated, but did not develop antibodies to other known mouse pathogens. Virus propagated in NCTC-1469 tissue culture produced enteric disease in suckling mice but not fatal diarrhea; the dams of these mice also developed antibodies to MHV and to the isolates. By complement fixation, single radial hemolysis, and quantal neutralization tests, we found the isolates antigenically most closely related to MHV-S, unilaterally related to MHV-JHM, and more distantly related to MHV-1, MHV-3, MHV-A59, and human coronavirus OC-43. We also studied cross-reactions among the murine and human coronaviruses in detail. Tissues of infected newborn mice were examined by light microscopy, thin-section electron microscopy, and frozen-section indirect immunofluorescence, revealing that viral antigen, virus particles, and pathological changes were limited to the intestinal tract. We have designated our isolates as MHV-S/CDC. Images PMID:222687

  8. Comparative study of two extraction methods for enteric virus recovery from sewage sludge by molecular methods.

    PubMed

    Schlindwein, A D; Simões, C M O; Barardi, C R M

    2009-07-01

    The aim of this study was to compare two nucleic acid extraction methods for the recovery of enteric viruses from activated sludge. Test samples were inoculated with human adenovirus (AdV), hepatitis A virus (HAV), poliovirus (PV) and rotavirus (RV) and were then processed by an adsorption-elution-precipitation method. Two extraction methods were used: an organic solvent-based method and a silica method. The organic-based method was able to recoup 20% of the AdV, 90% of the RV and 100% of both the PV and HAV from seeded samples. The silica method was able to recoup 1.8% of the AdV and 90% of the RV. These results indicate that the organic-based method is more suitable for detecting viruses in sewage sludge.

  9. Enteric virus infection risk from intrusion of sewage into a drinking water distribution network.

    PubMed

    Teunis, P F M; Xu, M; Fleming, K K; Yang, J; Moe, C L; Lechevallier, M W

    2010-11-15

    Contaminants from the soil surrounding drinking water distribution systems are thought to not enter the drinking water when sufficient internal pressure is maintained. Pressure transients may cause short intervals of negative pressure, and the soil near drinking water pipes often contains fecal material due to the proximity of sewage lines, so that a pressure event may cause intrusion of pathogens. This paper presents a risk model for predicting intrusion and dilution of viruses and their transport to consumers. Random entry and dilution of virus was simulated by embedding the hydraulic model into a Monte Carlo simulation. Special attention was given to adjusting for the coincidence of virus presence and use of tap water, as independently occurring short-term events within the longer interval that the virus is predicted to travel in any branch of the distribution system. The probability that a consumer drinks water contaminated with virus is small, but when this happens the virus concentration tends to be high and the risk of infection may be considerable. The spatial distribution of infection risk is highly heterogeneous. The presence of a chlorine residual reduces the infection risk.

  10. Three-Year Study To Assess Human Enteric Viruses in Shellfish

    PubMed Central

    Le Guyader, F.; Haugarreau, L.; Miossec, L.; Dubois, E.; Pommepuy, M.

    2000-01-01

    The main pathogenic enteric viruses able to persist in the environment, such as hepatitis A virus (HAV), Norwalk-like virus (NLV), enterovirus (EV), rotavirus (RV), and astrovirus (AV), were detected by reverse transcription-PCR and hybridization in shellfish during a 3-year study. Oyster samples (n = 108), occasionally containing bacteria, were less frequently contaminated, showing positivity for AV (17%), NLV (23%), EV (19%), and RV (27%), whereas mussel samples, collected in areas routinely impacted by human sewage, were more highly contaminated: AV (50%), HAV (13%), NLV (35%), EV (45%), and RV (52%). Sequences obtained from HAV and NLV amplicons showed a great variety of strains, especially for NLV (strains close to Mexico, Snow Mountain Agent, or Norwalk virus). Viral contamination was mainly observed during winter months, although there were some seasonal differences among the viruses. This first study of virus detection over a fairly long period of time suggests that routine analysis of shellfish by a molecular technique is feasible. PMID:10919776

  11. Molecular Detection and Genotyping of Male-Specific Coliphages by Reverse Transcription-PCR and Reverse Line Blot Hybridization

    PubMed Central

    Vinjé, Jan; Oudejans, Sjon J. G.; Stewart, Jill R.; Sobsey, Mark D.; Long, Sharon C.

    2004-01-01

    In recent years, there has been increased interest in the use of male-specific or F+ coliphages as indicators of microbial inputs to source waters. Sero- or genotyping of these coliphages can also be used for microbial source tracking (MST). Among the male-specific coliphages, the F+ RNA (FRNA) viruses are well studied, while little is known about the F+ DNA (FDNA) viruses. We have developed a reverse line blot hybridization (RLB) assay which allows for the simultaneous detection and genotyping of both FRNA as well as FDNA coliphages. These assays included a novel generic duplex reverse transcription-PCR (RT-PCR) assay for FRNA viruses as well as a generic PCR for FDNA viruses. The RT-PCR assays were validated by using 190 field and prototype strains. Subsequent DNA sequencing and phylogenetic analyses of RT-PCR products revealed the classification of six different FRNA clusters, including the well-established subgroups I through IV, and three different FDNA clusters, including one (CH) not previously described. Within the leviviruses, a potentially new subgroup (called JS) including strains having more than 40% nucleotide sequence diversity with the known levivirus subgroups (MS2 and GA) was identified. We designed subgroup-specific oligonucleotides that were able to genotype all nine (six FRNA, three FDNA) different clusters. Application of the method to a panel of 351 enriched phage samples from animal feces and wastewater, including known prototype strains (MS2, GA, Qβ, M11, FI, and SP for FRNA and M13, f1, and fd for FDNA), resulted in successful genotyping of 348 (99%) of the samples. In summary, we developed a novel method for standardized genotyping of F+ coliphages as a useful tool for large-scale MST studies. PMID:15466543

  12. Phenolic Compounds of Potato Peel Extracts: Their Antioxidant Activity and Protection against Human Enteric Viruses.

    PubMed

    Silva-BeltrÁn, Norma Patricia; Chaidez-Quiroz, Cristóbal; López-Cuevas, Osvaldo; Ruiz-Cruz, Saul; López-Mata, Marco A; Del-Toro-SÁnchez, Carmen Lizette; Marquez-Rios, Enrique; Ornelas-Paz, José de Jesús

    2017-02-28

    Potato peels (PP) contain several bioactive compounds. These compounds are known to provide human health benefits, including antioxidant and antimicrobial properties. In addition, these compounds could have effects on human enteric viruses that have not yet been reported. The objective of the present study was to evaluate the phenolic composition, antioxidant properties in the acidified ethanol extract (AEE) and water extract of PP, and the antiviral effects on the inhibition of Av-05 and MS2 bacteriophages, which were used as human enteric viral surrogates. The AEE showed the highest phenolic content and antioxidant activity. Chlorogenic and caffeic acids were the major phenolic acids. In vitro analysis indicated that PP had a strong antioxidant activity. A 3 h incubation with AEE at a concentration of 5 mg/ml was needed to reduce the PFU/ml (plaque-forming unit per unit volume) of Av-05 and MS2 by 2.8 and 3.9 log₁₀, respectively, in a dose-dependent manner. Our data suggest that PP has potential to be a source of natural antioxidants against enteric viruses.

  13. Propidium Monoazide Coupled with PCR Predicts Infectivity of Enteric Viruses in Swine Manure and Biofertilized Soil.

    PubMed

    Fongaro, Gislaine; Hernández, Marta; García-González, María Cruz; Barardi, Célia Regina Monte; Rodríguez-Lázaro, David

    2016-03-01

    The use of propidium monoazide (PMA) coupled with real-time PCR (RT-qPCR or qPCR for RNA or DNA viruses, respectively) was assessed to discriminate infectious enteric viruses in swine raw manure, swine effluent from anaerobic biodigester (AB) and biofertilized soils. Those samples were spiked either with infectious and heat-inactivated human adenovirus-2 (HAdV-2) or mengovirus (vMC0), and PMA-qPCR/RT-qPCR allowed discriminating inactivated viruses from the infective particles, with significant reductions (>99.9%). Then, the procedure was further assayed to evaluate the presence and stability of two non-cultivable viruses (porcine adenovirus and rotavirus A) in natural samples (swine raw manure, swine effluent from AB and biofertilized soils); it demonstrated viral inactivation during the storage period at 23 °C. As a result, the combination of PMA coupled to real-time PCR can be a promising alternative for prediction of viral infectivity in comparison to more labour-intensive and costly techniques such as animal or tissue-culture infectivity methods, and for those viruses that do not have currently available cell culture techniques.

  14. Detection of low levels of enteric viruses in metropolitan and airplane sewage.

    PubMed Central

    Shieh, Y S; Baric, R S; Sobsey, M D

    1997-01-01

    To detect less prevalent viruses, such as wild-type polioviruses in sewage from a highly immunized community, a method was developed to efficiently recover viruses and remove PCR inhibitors. The method consisted of initial separation of solids from liquid, followed by solvent extractions, polyethylene glycol precipitations, Sephadex G-200 chromatography, and guanidinium isothiocyanate (GIT) extraction. To elute viruses from the separated solids, 0.5 M threonine (pH 7.5) was as efficient as 3% beef extract but conferred no PCR inhibition. In samples that were concentrated approximately 1,000-fold, 21% of the initially seeded viruses were recovered. When poliovirus type 3 (PV3) Sabin strain at low levels and PV1 LSc strain at high levels were seeded in raw sewage, PV3 was specifically detected in the final sample concentrates at sensitivities of 14 PFU by direct PCR and 0.7 PFU by GIT extraction-PCR. While applying the method to international airplane sewage, which contains high levels of solids as well as commercial sanitizers, 44% (7 of 16) of the samples were found to harbor enteroviruses by both cell culture infectivity and pan-enterovirus PCR analyses. Nucleotide sequencing of the PCR products revealed that multiple enterovirus genotypes were amplified from each final sewage concentrate, whereas the fewer virus genotypes detected by cell culture infectivity were probably the better growing strains. By this method, we demonstrated that air travel may contribute to the intercontinental dissemination of enteric pathogens. PMID:9361427

  15. Inactivation of internalized and surface contaminated enteric viruses in green onions.

    PubMed

    Hirneisen, Kirsten A; Kniel, Kalmia E

    2013-09-02

    With increasing outbreaks of gastroenteritis associated with produce, it is important to assess interventions to reduce the risk of illness. UV, ozone and high pressure are non-thermal processing technologies that have potential to inactivate human pathogens on produce and allow the retention of fresh-like organoleptic properties. The objective of this study was to determine if UV, ozone, and high pressure are effective technologies compared to traditional chlorine spray on green onions to reduce enteric viral pathogens and to determine the effect of location of the virus (surface or internalized) on the efficacy of these processes. Mature green onion plants were inoculated with murine norovirus (MNV), hepatitis A virus (HAV) and human adenovirus type 41 (Ad41) either on the surface through spot inoculation or through inoculating contaminated hydroponic solution allowing for uptake of the virus into the internal tissues. Inoculated green onions were treated with UV (240 mJ s/cm(2)), ozone (6.25 ppm for 10 min), pressure (500 MPa, for 5 min at 20°C), or sprayed with calcium hypochlorite (150 ppm, 4°C). Viral inactivation was determined by comparing treated and untreated inoculated plants using cell culture infectivity assays. Processing treatments were observed to greatly affect viral inactivation. Viral inactivation for all three viruses was greatest after pressure treatment and the lowest inactivation was observed after chlorine and UV treatment. Both surface inoculated viruses and viruses internalized in green onions were inactivated to some extent by these post-harvest processing treatments. These results suggest that ozone and high pressure processes aimed to reduce the level of microbial contamination of produce have the ability to inactivate viruses if they become localized in the interior portions of produce.

  16. Myxoma and vaccinia viruses exploit different mechanisms to enter and infect human cancer cells

    SciTech Connect

    Villa, Nancy Y.; Bartee, Eric; Mohamed, Mohamed R.; Rahman, Masmudur M.; Barrett, John W.; McFadden, Grant

    2010-06-05

    Myxoma (MYXV) and vaccinia (VACV) viruses have recently emerged as potential oncolytic agents that can infect and kill different human cancer cells. Although both are structurally similar, it is unknown whether the pathway(s) used by these poxviruses to enter and cause oncolysis in cancer cells are mechanistically similar. Here, we compared the entry of MYXV and VACV-WR into various human cancer cells and observed significant differences: 1 - low-pH treatment accelerates fusion-mediated entry of VACV but not MYXV, 2 - the tyrosine kinase inhibitor genistein inhibits entry of VACV, but not MYXV, 3 - knockdown of PAK1 revealed that it is required for a late stage event downstream of MYXV entry into cancer cells, whereas PAK1 is required for VACV entry into the same target cells. These results suggest that VACV and MYXV exploit different mechanisms to enter into human cancer cells, thus providing some rationale for their divergent cancer cell tropisms.

  17. Myxoma and vaccinia viruses exploit different mechanisms to enter and infect human cancer cells.

    PubMed

    Villa, Nancy Y; Bartee, Eric; Mohamed, Mohamed R; Rahman, Masmudur M; Barrett, John W; McFadden, Grant

    2010-06-05

    Myxoma (MYXV) and vaccinia (VACV) viruses have recently emerged as potential oncolytic agents that can infect and kill different human cancer cells. Although both are structurally similar, it is unknown whether the pathway(s) used by these poxviruses to enter and cause oncolysis in cancer cells are mechanistically similar. Here, we compared the entry of MYXV and VACV-WR into various human cancer cells and observed significant differences: 1--low-pH treatment accelerates fusion-mediated entry of VACV but not MYXV, 2--the tyrosine kinase inhibitor genistein inhibits entry of VACV, but not MYXV, 3--knockdown of PAK1 revealed that it is required for a late stage event downstream of MYXV entry into cancer cells, whereas PAK1 is required for VACV entry into the same target cells. These results suggest that VACV and MYXV exploit different mechanisms to enter into human cancer cells, thus providing some rationale for their divergent cancer cell tropisms.

  18. Real-time fluorescence loop-mediated isothermal amplification for the diagnosis of hemorrhagic enteritis virus.

    PubMed

    Liu, Xuemei; Li, Yuhao; Xu, Chenggang; Qin, Jianru; Hao, Jianyong; Feng, Min; Tan, Liqiang; Jia, Weixin; Liao, Ming; Cao, Weisheng

    2014-04-01

    Suspected cases of hemorrhagic enteritis associated with hemorrhagic enteritis virus (HEV) are becoming more frequent among yellow chickens in the Guangdong Province of China. In this study, we have developed a one-step, ecumenical, real-time fluorescence loop-mediated isothermal amplification (RealAmp) assay for the rapid diagnosis of HEV. The RealAmp assay was performed at 63°C and reduced the assay time to 15min, using a simple and portable device, the ESE-Quant Tube Scanner. The detection limit of DNA was 1fg/μl, and the detection was specific only to HEV. We also used nested PCR to evaluate the application of the RealAmp assay. The coincidence rate of the two methods was 100%. Our data indicated that the RealAmp assay provides a sensitive, specific, and user-friendly diagnostic tool for the identification and quantification of HEV for field diagnosis and in laboratory research.

  19. Adaptation and growth kinetics study of an Indian isolate of virulent duck enteritis virus in Vero cells.

    PubMed

    Aravind, S; Kamble, Nitin M; Gaikwad, Satish S; Shukla, Sanjeev Kumar; Dey, Sohini; Mohan, C Madhan

    2015-01-01

    Duck virus enteritis, also known as duck plague, is a viral infection of ducks caused by duck enteritis virus (DEV). The control of the disease is mainly done by vaccination with chicken embryo adapted live virus that is known to be poorly immunogenic and elicits only partial protection. Further, the embryo propagated vaccine virus pose a threat of harboring other infectious agents. Seeing these limitations, the present study reports for the first time regarding propagation and adaptation of a virulent Indian isolate of duck enteritis virus in Vero cell line. In this study isolation of an outbreak virus from Kerala state was done in chicken embryo fibroblast cell culture (CEF). Then adapted the DEV isolate in the Vero cell line. The characteristic cytopathic effects (CPE) of clumping and fusion of Vero cells were observed starting from the 7th passage onwards. The presence of the virus and its multiplication in Vero cells was confirmed by detection of viral specific DNA and antigen by using polymerase chain reaction (PCR) and indirect immuno fluorescent assay (IIFA), respectively. PCR detection of DEV using self designed primers for US4 (gD) and UL30 (DNA Polymerase) gene has been reported for the in the present study. The kinetics of DEV in Vero cells revealed a maximum infectivity titer of 10(5.6) TCID 50/ml after 48hr of viral infection. Compared to chicken embryo adapted DVE vaccine virus, the Vero cell culture system is free from other infectious agents. So it will be a good candidate for cultivation and propagation of duck enteritis virus vaccine strain. Further research studies are suggested to explore the feasibility of utilizing this Vero cell culture adapted DEV isolate for developing an attenuated vaccine virus against duck virus enteritis.

  20. Characterisation of bovine viral diarrhoea virus (BVDV) isolates from an outbreak with haemorrhagic enteritis and severe pneumonia.

    PubMed

    Yeşilbağ, Kadir; Förster, Christine; Ozyiğit, M Ozgür; Alpay, Gizem; Tuncer, Pelin; Thiel, Heinz-Jürgen; König, Matthias

    2014-02-21

    During 2007 a disease outbreak occurred in cattle in the Marmara region of western Turkey characterised by severe pneumonia and haemorrhagic enteritis in calves. Cases from three farms at different locations were examined and bovine viral diarrhoea virus (BVDV) isolated in all cases. Phylogenetic characterisation of the virus isolates allocated them in a new cluster tentatively named as BVDV-1r.

  1. THE USE OF RT-PCR FOR THE DETECTION OF ENTERIC VIRUSES IN PRAIRIE SURFACE DRINKING WATER SUPPLIES

    EPA Science Inventory

    Concerns over the microbial safety of drinking water supplies have focused on bacteria and parasites while the occurrence of pathogenic waterborne viruses have been largely ignored. In fact, water supplies are not routinely monitored for human enteric viruses. This is despite t...

  2. Use of propidium monoazide in reverse transcriptase PCR to distinguish between infectious and noninfectious enteric viruses in water samples

    EPA Science Inventory

    Human enteric viruses can be present in untreated and inadequately treated drinking water. Molecular methods, such as the reverse transcriptase PCR (RT-PCR), can detect viral genomes in a few hours, but they cannot distinguish between infectious and noninfectious viruses. Since o...

  3. HN gene c-terminal extension of Newcastle disease virus is not the determinant of the enteric tropism

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) plays an important role in virus pathogenicity and tissue tropism. Sequence analysis revealed that the HN gene of many asymptomatic enteric NDV strains encodes a larger open reading frame (616 amino acids, aa) with additio...

  4. Round-robin comparison of methods for the detection of human enteric viruses in lettuce.

    PubMed

    Le Guyader, Françoise S; Schultz, Anna-Charlotte; Haugarreau, Larissa; Croci, Luciana; Maunula, Leena; Duizer, Erwin; Lodder-Verschoor, Froukje; von Bonsdorff, Carl-Henrik; Suffredini, Elizabetha; van der Poel, Wim M M; Reymundo, Rosanna; Koopmans, Marion

    2004-10-01

    Five methods that detect human enteric virus contamination in lettuce were compared. To mimic multiple contaminations as observed after sewage contamination, artificial contamination was with human calicivirus and poliovirus and animal calicivirus strains at different concentrations. Nucleic acid extractions were done at the same time in the same laboratory to reduce assay-to-assay variability. Results showed that the two critical steps are the washing step and removal of inhibitors. The more reliable methods (sensitivity, simplicity, low cost) included an elution/concentration step and a commercial kit. Such development of sensitive methods for viral detection in foods other than shellfish is important to improve food safety.

  5. Detection of enteric viruses in oysters by using the polymerase chain reaction.

    PubMed Central

    Atmar, R L; Metcalf, T G; Neill, F H; Estes, M K

    1993-01-01

    A procedure for the detection of enteric viral nucleic acid in oysters by the polymerase chain reaction was developed. Known quantities of poliovirus type 1 were seeded into oysters. Virus was extracted and concentrated by using organic flocculation and polyethylene glycol precipitation. Inhibitors of reverse transcription-polymerase chain reaction were present in the oyster extracts, preventing amplification of target viral nucleic acid. The use of cetyltrimethylammonium bromide precipitation sufficiently removed inhibitors to allow detection of as few as 10 PFU of poliovirus. Norwalk virus also could be detected after being seeded into oysters. This methodology may be useful for the detection of these and other shellfish-borne viral pathogens. Images PMID:8382024

  6. Abundance and Distribution of Enteric Bacteria and Viruses in Coastal and Estuarine Sediments—a Review

    PubMed Central

    Hassard, Francis; Gwyther, Ceri L.; Farkas, Kata; Andrews, Anthony; Jones, Vera; Cox, Brian; Brett, Howard; Jones, Davey L.; McDonald, James E.; Malham, Shelagh K.

    2016-01-01

    The long term survival of fecal indicator organisms (FIOs) and human pathogenic microorganisms in sediments is important from a water quality, human health and ecological perspective. Typically, both bacteria and viruses strongly associate with particulate matter present in freshwater, estuarine and marine environments. This association tends to be stronger in finer textured sediments and is strongly influenced by the type and quantity of clay minerals and organic matter present. Binding to particle surfaces promotes the persistence of bacteria in the environment by offering physical and chemical protection from biotic and abiotic stresses. How bacterial and viral viability and pathogenicity is influenced by surface attachment requires further study. Typically, long-term association with surfaces including sediments induces bacteria to enter a viable-but-non-culturable (VBNC) state. Inherent methodological challenges of quantifying VBNC bacteria may lead to the frequent under-reporting of their abundance in sediments. The implications of this in a quantitative risk assessment context remain unclear. Similarly, sediments can harbor significant amounts of enteric viruses, however, the factors regulating their persistence remains poorly understood. Quantification of viruses in sediment remains problematic due to our poor ability to recover intact viral particles from sediment surfaces (typically <10%), our inability to distinguish between infective and damaged (non-infective) viral particles, aggregation of viral particles, and inhibition during qPCR. This suggests that the true viral titre in sediments may be being vastly underestimated. In turn, this is limiting our ability to understand the fate and transport of viruses in sediments. Model systems (e.g., human cell culture) are also lacking for some key viruses, preventing our ability to evaluate the infectivity of viruses recovered from sediments (e.g., norovirus). The release of particle-bound bacteria and

  7. Abundance and Distribution of Enteric Bacteria and Viruses in Coastal and Estuarine Sediments-a Review.

    PubMed

    Hassard, Francis; Gwyther, Ceri L; Farkas, Kata; Andrews, Anthony; Jones, Vera; Cox, Brian; Brett, Howard; Jones, Davey L; McDonald, James E; Malham, Shelagh K

    2016-01-01

    The long term survival of fecal indicator organisms (FIOs) and human pathogenic microorganisms in sediments is important from a water quality, human health and ecological perspective. Typically, both bacteria and viruses strongly associate with particulate matter present in freshwater, estuarine and marine environments. This association tends to be stronger in finer textured sediments and is strongly influenced by the type and quantity of clay minerals and organic matter present. Binding to particle surfaces promotes the persistence of bacteria in the environment by offering physical and chemical protection from biotic and abiotic stresses. How bacterial and viral viability and pathogenicity is influenced by surface attachment requires further study. Typically, long-term association with surfaces including sediments induces bacteria to enter a viable-but-non-culturable (VBNC) state. Inherent methodological challenges of quantifying VBNC bacteria may lead to the frequent under-reporting of their abundance in sediments. The implications of this in a quantitative risk assessment context remain unclear. Similarly, sediments can harbor significant amounts of enteric viruses, however, the factors regulating their persistence remains poorly understood. Quantification of viruses in sediment remains problematic due to our poor ability to recover intact viral particles from sediment surfaces (typically <10%), our inability to distinguish between infective and damaged (non-infective) viral particles, aggregation of viral particles, and inhibition during qPCR. This suggests that the true viral titre in sediments may be being vastly underestimated. In turn, this is limiting our ability to understand the fate and transport of viruses in sediments. Model systems (e.g., human cell culture) are also lacking for some key viruses, preventing our ability to evaluate the infectivity of viruses recovered from sediments (e.g., norovirus). The release of particle-bound bacteria and

  8. Molecular epidemiology of enteric viruses in children with sporadic gastroenteritis in Valencia, Venezuela.

    PubMed

    González, Germán G; Liprandi, Ferdinando; Ludert, Juan E

    2011-11-01

    The epidemiology and clinical symptoms in infants and young children with acute sporadic viral gastroenteritis due to viral etiologies other than rotaviruses have not been studied thoroughly in developing countries. Fecal specimens from 480 children <5 years of age who were admitted to a large children's hospital in the city of Valencia, Venezuela, with acute diarrhea during January to December 2003 were collected and screened by ELISA and RT-PCR for rotavirus, adenovirus, norovirus, sapovirus, and astrovirus. Viral isolates were partially characterized by phylogenetic analysis. Norovirus viral load was determined by qRT-PCR. Viruses were identified in 205 (43%) of the 480 stool samples collected. Rotavirus was the virus detected most frequently (21%), followed by norovirus (13%), adenovirus (5%), sapovirus (3%), and astrovirus (2%). Viral infection rates were highest in the 6- to 11-month-old group (49%) and lowest in children >24 months old. Norovirus GII was more prevalent (90%) than GI (10%). Enteric adenovirus (serotypes 40/41) was present in 43% of the adenovirus-positive samples. Rotavirus infection caused more severe clinical symptoms than the other viruses detected, with more vomiting (84%) and dehydration (11%) that led to hospital admission of 20% of the children with acute gastroenteritis. Rotavirus and norovirus showed marked and opposite seasonal patterns. No association was observed between disease severity and viral load in children infected with norovirus. These results not only confirm the impact of rotavirus infection in Venezuela but also indicate that other enteric viruses, especially noroviruses, contribute significantly to sporadic acute gastroenteritis and to the burden of disease.

  9. Release of infectious human enteric viruses by full-scale wastewater utilities.

    PubMed

    Simmons, Fredrick James; Xagoraraki, Irene

    2011-06-01

    In the United States, infectious human enteric viruses are introduced daily into the environment through the discharge of treated water and the digested sludge (biosolids). In this study, a total of 30 wastewater and 6 biosolids samples were analyzed over five months (May-September 2008-2009) from five full-scale wastewater treatment plants (WWTPs) in Michigan using real-time PCR and cell culture assays. Samples were collected from four different locations at each WWTP (influent, pre-disinfection, post-disinfection and biosolids) using the 1MDS electropositive cartridge filter. Adenovirus (HAdV), enterovirus (EV) and norovirus genogroup II (NoV GGII) were detected in 100%, 67% and 10%, respectively of the wastewater samples using real-time PCR. Cytopathic effect (CPE) was present in 100% of the cell culture samples for influent, pre- and post-disinfection and biosolids with an average log concentration of 4.1 (2.9-4.7, range) 1.1 (0.0-2.3, range) and 0.5 (0.0-1.6, range) MPN/100 L and 2.1 (0.5-4.1) viruses/g, respectively. A significant log reduction in infectious viruses throughout the wastewater treatment process was observed at an average 4.2 (1.9-5.0, range) log units. A significant difference (p-value <0.05) was observed using real-time PCR data for HAdV but not for EV (p-value >0.05) removal in MBR as compared to conventional treatment. MBR treatment was able to achieve an additional 2 and 0.5 log reduction of HAdV and EV, respectively. This study has demonstrated the release of infectious enteric viruses in the final effluent and biosolids of wastewater treatment into the environment.

  10. Analysis of experimental mink enteritis virus infection in mink: in situ hybridization, serology, and histopathology.

    PubMed Central

    Uttenthal, A; Larsen, S; Lund, E; Bloom, M E; Storgård, T; Alexandersen, S

    1990-01-01

    Strand-specific hybridization probes were used in in situ hybridization studies to localize cells containing mink enteritis virus (MEV) virion DNA or MEV replicative-form DNA and mRNA. Following the experimental MEV infection of 3-month-old unvaccinated mink, a significant increase in serum antibodies to MEV was detected at postinfection day (PID) 6, 2 days after the onset of fecal shedding of virus. Prior to the appearance of virus in feces, viral DNA could be detected in the mesenteric lymph node and intestine. The largest percentage of cells positive for virion DNA was 10% and was detected in the intestine on PID 6. However, replication of the virus apparently peaked at PID 4. The number of MEV replicative-form DNA and mRNA molecules was found to be approximately 250,000 copies per infected lymph node cell or crypt epithelial cell. The localization, levels, and time course of viral replication have important implications for the pathogenesis of MEV-induced disease. The data presented on MEV are correlated with earlier results on the other mink parvovirus, Aleutian mink disease parvovirus, and a possible explanation for the remarkable differences in pathogenesis of disease caused by these two parvoviruses is discussed. Images PMID:2159543

  11. Acute gastroenteritis and enteric viruses in hospitalised children in southern Brazil: aetiology, seasonality and clinical outcomes.

    PubMed

    Raboni, Sonia Maria; Damasio, Guilherme Augusto Costa; Ferreira, Carla E O; Pereira, Luciane A; Nogueira, Meri B; Vidal, Luine R; Cruz, Cristina R; Almeida, Sergio M

    2014-07-01

    Viral acute gastroenteritis (AG) is a significant cause of hospitalisation in children younger than five years. Group A rotavirus (RVA) is responsible for 30% of these cases. Following the introduction of RVA immunisation in Brazil in 2006, a decreased circulation of this virus has been observed. However, AG remains an important cause of hospitalisation of paediatric patients and only limited data are available regarding the role of other enteric viruses in these cases. We conducted a prospective study of paediatric patients hospitalised for AG. Stool samples were collected to investigate human adenovirus (HAdV), RVA, norovirus (NoV) and astrovirus (AstV). NoV typing was performed by nucleotide sequencing and phylogenetic analysis. From the 225 samples tested, 60 (26%) were positive for at least one viral agent. HAdV, NoV, RVA and AstV were detected in 16%, 8%, 6% and 0% of the samples, respectively. Mixed infections were found in nine patients: HAdV/RVA (5), HAdV/NoV (3) and HAdV/NoV/RVA (1). The frequency of fever and lymphocytosis was significantly higher in virus-infected patients. Phylogenetic analysis of NoV indicated that all of these viruses belonged to genotype GII.4. The significant frequency of these pathogens in patients with AG highlights the need to routinely implement laboratory investigations.

  12. Large scale survey of enteric viruses in river and waste water underlines the health status of the local population.

    PubMed

    Prevost, B; Lucas, F S; Goncalves, A; Richard, F; Moulin, L; Wurtzer, S

    2015-06-01

    Although enteric viruses constitute a major cause of acute waterborne diseases worldwide, environmental data about occurrence and viral load of enteric viruses in water are not often available. In this study, enteric viruses (i.e., adenovirus, aichivirus, astrovirus, cosavirus, enterovirus, hepatitis A and E viruses, norovirus of genogroups I and II, rotavirus A and salivirus) were monitored in the Seine River and the origin of contamination was untangled. A total of 275 water samples were collected, twice a month for one year, from the river Seine, its tributaries and the major WWTP effluents in the Paris agglomeration. All water samples were negative for hepatitis A and E viruses. AdV, NVGI, NVGII and RV-A were the most prevalent and abundant populations in all water samples. The viral load and the detection frequency increased significantly between the samples collected the most upstream and the most downstream of the Paris urban area. The calculated viral fluxes demonstrated clearly the measurable impact of WWTP effluents on the viral contamination of the Seine River. The viral load was seasonal for almost all enteric viruses, in accordance with the gastroenteritis recordings provided by the French medical authorities. These results implied the existence of a close relationship between the health status of inhabitants and the viral contamination of WWTP effluents and consequently surface water contamination. Subsequently, the regular analysis of wastewater could serve as a proxy for the monitoring of the human viruses circulating in both a population and surface water.

  13. Inactivation of enteric microorganisms with chemical disinfectants, UV irradiation and combined chemical/UV treatments.

    PubMed

    Koivunen, J; Heinonen-Tanski, H

    2005-04-01

    The relative disinfection efficiencies of peracetic acid (PAA), hydrogen peroxide (H2O2) and sodium hypochlorite (NaOCl) against Escherichia coli, Enterococcus faecalis, Salmonella enteritidis and coliphage MS2 virus were studied in laboratory-scale experiments. This study also evaluated the efficiency of combined PAA/ultraviolet irradiation (UV) and H2O2/UV treatments to determine if the microbial inactivation was synergistic. Microbial cultures were added into a synthetic wastewater-like test medium and treated by chemical disinfectants with a 10 min contact time, UV irradiation or the combination of chemical and UV treatments. A peracetic acid dose of 3 mg/l resulted in approximately 2-3 log enteric bacterial reductions, whereas 7-15 mg/l PAA was needed to achieve 1-1.5 log coliphage MS2 reductions. Doses of 3-150 mg/l hydrogen peroxide achieved below 0.2 log microbial reductions. Sodium hypochlorite treatments caused 0.3-1 log microbial reductions at an 18 mg/l chlorine dose, while 2.6 log reductions of E. faecalis were achieved at a 12 mg/l chlorine dose. The results indicate that PAA could represent a good alternative to chlorine compounds in disinfection procedures, especially in wastewaters containing easily oxidizable organic matter. Hydrogen peroxide is not an efficient disinfectant against enteric microorganisms in wastewaters. The combined PAA/UV disinfection showed increased disinfection efficiency and synergistic benefits with all the enteric bacteria tested but lower synergies for the coliphage MS2. This suggests that this method could improve the efficiency and reliability of disinfection in wastewater treatment plants. The combined H2O2/UV disinfection only slightly influenced the microbial reductions compared to UV treatments and showed some antagonism and no synergies.

  14. A survey of North American migratory waterfowl for duck plague (duck virus enteritis) virus

    USGS Publications Warehouse

    Brand, Christopher J.; Docherty, Douglas E.

    1984-01-01

    A survey of migratory waterfowl for duck plague (DP) virus was conducted in the Mississippi and Central flyways during 1982 and in the Atlantic and Pacific flyways during 1983. Cloacal and pharyngeal swabs were collected from 3,169 migratory waterfowl in these four flyways, principally mallards (Anas platyrhynchos L.), black ducks (Anas rubripes Brewster), and pintails (Anas acuta L). In addition 1,033 birds were sampled from areas of recurrent DP outbreaks among nonmigratory and captive waterfowl, and 590 from Lake Andes National Wildlife Refuge, the site of the only known major DP outbreak in migratory waterfowl. Duck plague virus was not found in any of the samples. Results support the hypothesis that DP is not established in North American migratory waterfowl as an enzootic disease.

  15. Assessment and risk modeling of airborne enteric viruses emitted from wastewater reused for irrigation.

    PubMed

    Courault, D; Albert, I; Perelle, S; Fraisse, A; Renault, P; Salemkour, A; Amato, P

    2017-08-15

    Reclamation of wastewater (WW) for irrigation, after treatment represents a challenge that could alleviate pressure on water resources and address the increasing demand for agriculture. However, the risks to human health must be assessed, particularly those related to human enteric viruses that resist standard treatments in most wastewater treatment plants (WWTP). The risks associated with exposure to viral bioaerosols near WWTP and near agricultural plots irrigated with WW are poorly documented. The objectives of this study were to 1) better characterize human enteric viruses found in bioaerosols near a "standard WWTP" and over fields irrigated with treated WW and 2) propose a numeric model to assess the health risk to populations located close to the irrigated areas, with particular attention to norovirus, which is responsible for most viral gastroenteritis in France. Water and air samples were collected at various locations in the largest French WW-irrigated site near Clermont-Ferrand, at the WWTP entrance and after treatment, in the air above activated sludge basins, and above fields irrigated with WW. Various enteric viruses were found in the water samples collected both before and after treatment. Norovirus was the most abundant with >10e4 genome copies/l (GC/L) before treatment and ~10e3 GC/L after treatment. Low quantities (<10e3GC/m(3)) were detected in the air above active sludge pools and irrigated plots. Hepatitis E virus was detected in all sampled compartments. A quantitative microbial risk assessment (QMRA) approach, including a simplified atmospheric dispersion model, allowed assessment of norovirus infection risk. The Bayesian QMRA approach considered wind speed measurements over 21years, and the variability and uncertainty of all measurements throughout the chain up to the risk. The probability of infection within one year for the most exposed WWTP employees was >10e-4 for strong wind speed (≥3m/s) and a constant emission rate of 8e3 GC/m(3)/s

  16. Diversity of Somatic Coliphages in Coastal Regions with Different Levels of Anthropogenic Activity in São Paulo State, Brazil ▿

    PubMed Central

    Burbano-Rosero, E. M.; Ueda-Ito, M.; Kisielius, J. J.; Nagasse-Sugahara, T. K.; Almeida, B. C.; Souza, C. P.; Markman, C.; Martins, G. G.; Albertini, L.; Rivera, I. N. G.

    2011-01-01

    Bacteriophages are the most abundant and genetically diverse viruses on Earth, with complex ecology in both quantitative and qualitative terms. Somatic coliphages (SC) have been reported to be good indicators of fecal pollution in seawater. This study focused on determining the concentration of SC and their diversity by electron microscopy of seawater, plankton, and bivalve samples collected at three coastal regions in São Paulo, Brazil. The SC counts varied from <1 to 3.4 × 103 PFU/100 ml in seawater (73 samples tested), from <1 to 4.7 × 102 PFU/g in plankton (46 samples tested), and from <1 to 2.2 × 101 PFU/g in bivalves (11 samples tested). In seawater samples, a relationship between the thermotolerant coliforms and Escherichia coli and SC was observed at the three regions (P = 0.0001) according to the anthropogenic activities present at each region. However, SC were found in plankton samples from three regions: Baixada Santista (17/20), Canal de São Sebastião (6/14), and Ubatuba (3/12). In seawater samples collected from Baixada Santista, four morphotypes were observed: A1 (4.5%), B1 (50%), C1 (36.4%), and D1 (9.1%). One coliphage, Siphoviridae type T1, had the longest tail: between 939 and 995 nm. In plankton samples, Siphoviridae (65.8%), Podoviridae (15.8%), Microviridae (15.8%), and Myoviridae (2.6%) were found. In bivalves, only the morphotype B1 was observed. These SC were associated with enteric hosts: enterobacteria, E. coli, Proteus, Salmonella, and Yersinia. Baixada Santista is an area containing a high level of fecal pollution compared to those in the Canal de São Sebastião and Ubatuba. This is the first report of coliphage diversity in seawater, plankton, and bivalve samples collected from São Paulo coastal regions. A better characterization of SC diversity in coastal environments will help with the management and evaluation of the microbiological risks for recreation, seafood cultivation, and consumption. PMID:21531842

  17. Bacillus subtilis and surfactin inhibit the transmissible gastroenteritis virus entering intestinal epithelial cells.

    PubMed

    Wang, Xiaoqing; Hu, Weiwei; Zhu, Liqi; Yang, Qian

    2017-03-07

    Intestinal epithelial cells are the targets for transmissible gastroenteritis virus (TGEV) infection. It is urgently to develop a novel candidate against TGEV entry. Bacillus subtilis is a probiotics with excellent anti-microorganism properties, and one of its secretions, surfactin, has been regarded as the versatile weapons for most plant pathogens, especially for the enveloped virus. We demonstrate for the first time that Bacillus subtilis OKB105 and its surfactin can effectively inhibit one animal coronavirus, TGEV, entering the intestinal porcine epithelial cell line (IPEC-J2). Then, several different experiments were performed to seek for the might mechanisms. The plaque assays showed that surfactant could reduce the plaque generation of TGEV in a dose dependent manner. Meanwhile, the after incubated with TGEV for 1.5 h, Bacillus subtilis could attach TGEV particles to their surface so that the number of virus to bind to the host cells was declined. Furthermore, our data showed that the inhibition of Bacillus subtilis was closely related to the competition with TGEV for the viral-entry receptors, including epidermal growth factor receptor (EGFR) and aminopopeptidase N (APN) protein. In addition, Western blotting and apoptosis analysis indicated that Bacillus subtilis could enhance the resistance of IPEC-J2 cells by up regulating the expression of TLR-6 and reducing the percentage of apoptotic cells. Taken together, our results suggest that Bacillus subtilis OKB105 and its surfactin can antagonize TGEV entry in vitro and may serve as promising new candidates for TGEV prevention.

  18. An enteric virus can replace the beneficial function of commensal bacteria

    PubMed Central

    Kernbauer, Elisabeth; Ding, Yi; Cadwell, Ken

    2014-01-01

    Intestinal microbial communities have profound effects on host physiology1. Whereas the symbiotic contribution of commensal bacteria is well established, the role of eukaryotic viruses that are present in the gastrointestinal tract under homeostatic conditions is undefined2,3. Here, we demonstrate that a common enteric RNA virus can replace the beneficial function of commensal bacteria in the intestine. Murine norovirus (MNV) infection of germfree or antibiotics-treated mice restored intestinal morphology and lymphocyte function without inducing overt inflammation and disease. The presence of MNV also suppressed an expansion of group 2 innate lymphoid cells (ILCs) observed in the absence of bacteria, and induced transcriptional changes in the intestine associated with immune development and type I interferon (IFN) signaling. Consistent with this observation, the IFNα receptor was essential for the ability of MNV to compensate for bacterial depletion. Importantly, MNV infection offset the deleterious effect of antibiotics-treatment in models of intestinal injury and pathogenic bacterial infection. These data indicate that eukaryotic viruses have the capacity to support intestinal homeostasis and shape mucosal immunity akin to commensal bacteria. PMID:25409145

  19. Enteric Viruses of Humans and Animals in Aquatic Environments: Health Risks, Detection, and Potential Water Quality Assessment Tools

    PubMed Central

    Fong, Theng-Theng; Lipp, Erin K.

    2005-01-01

    Waterborne enteric viruses threaten both human and animal health. These pathogens are host specific and cause a wide range of diseases and symptoms in humans or other animals. While considerable research has documented the risk of enteric viruses to human health from contact with contaminated water, the current bacterial indicator-based methods for evaluation of water quality are often ineffectual proxies for pathogenic viruses. Additionally, relatively little work has specifically investigated the risk of waterborne viruses to animal health, and this risk currently is not addressed by routine water quality assessments. Nonetheless, because of their host specificity, enteric viruses can fulfill a unique role both for assessing health risks and as measures of contamination source in a watershed, yet the use of animal, as well as human, host-specific viruses in determining sources of fecal pollution has received little attention. With improved molecular detection assays, viruses from key host groups can be targeted directly using PCR amplification or hybridization with a high level of sensitivity and specificity. A multispecies viral analysis would provide needed information for controlling pollution by source, determining human health risks based on assessments of human virus loading and exposure, and determining potential risks to production animal health and could indicate the potential for the presence of other zoonotic pathogens. While there is a need to better understand the prevalence and environmental distribution of nonhuman enteric viruses, the development of improved methods for specific and sensitive detection will facilitate the use of these microbes for library-independent source tracking and water quality assessment tools. PMID:15944460

  20. Enteric viruses and adenovirus diversity in waters from 2016 Olympic venues.

    PubMed

    Staggemeier, Rodrigo; Heck, Tatiana M S; Demoliner, Meriane; Ritzel, Rute G F; Röhnelt, Nicole M S; Girardi, Viviane; Venker, Carolina A; Spilki, Fernando R

    2017-02-06

    Rio de Janeiro's inner and coastal waters are heavily impacted by human sewage pollution for decades. Enteric viruses, including human adenoviruses (HAdV), human enterovirus (EV), group A rotavirus (RV) and hepatitis A virus (HAV) are more likely to be found in contaminated surface waters. The present work aimed to assess the frequency and loads of EV, HAdV-C and -F species, RV and HAV in sand and water samples from venues used during the 2016 Summer Olympics and by tourists attending the event. Sixteen monthly collections were carried out from March 2015 to July 2016 in 12 different sites from Rio de Janeiro, Brazil. Total and thermotolerant coliform counting was performed along molecular detection of virus was performed using quantitative polymerase chain reaction (qPCR). Analyses of all samples were further investigated by integrated cell culture PCR to check about the presence of HAdV infectious virus particles. The results show that 95.9% of water samples showed contamination with at least one type of virus. Regarding the viruses individually (% for water and sand respectively): HAdV-C (93.1%-57.8%), HAdV-F (25.3%-0%), RV (12.3%-4.4%), EV (26.7%-8.8%) and HAV (0%). The viral loads ranged from 10(3)gc/L up to 10(9)gc/L (water), and 10(3)gc/g to 10(6)gc/g (sand). In the phylogenetic tree, were classified into four main clusters, referring to species C, D, F and BAdV. And up to 90% of sites studied presented at least once presence of infectious HAdV-C. The most contaminated points were the Rodrigo de Freitas Lagoon, where Olympic rowing took place, and the Marina da Glória, the starting point for the sailing races, demonstrating serious problem of fecal contamination of water resources and threatens the health of Olympic athletes, tourists and residents.

  1. White spot syndrome virus enters crayfish hematopoietic tissue cells via clathrin-mediated endocytosis.

    PubMed

    Huang, Jiajun; Li, Fang; Wu, Junjun; Yang, Feng

    2015-12-01

    White spot syndrome virus (WSSV) is a major pathogen of aquacultured shrimp. However, the mechanism of its entry remains poorly understood. In this study, by analyzing the internalization of WSSV using crayfish hematopoietic tissue (HPT) cells, we showed that WSSV virions were engulfed by cell membrane invaginations sharing the features of clathrin-coated pits and then internalized into coated cytoplasmic vesicles. Further investigation indicated that WSSV internalization was significantly inhibited by chlorpromazine (CPZ) but not genistein. The internalized virions were colocalized with endogenous clathrin as well as transferrin which undergoes clathrin-dependent uptake. Preventing endosome acidification by ammonium chloride (NH4Cl) or chloroquine (CQ) dramatically reduced WSSV entry as well. Moreover, disturbance of dynamin activity or depletion of membrane cholesterol also blocked WSSV uptake. These data indicate that WSSV enters crayfish HPT cells via clathrin-mediated endocytosis in a pH-dependent manner, and membrane cholesterol as well as dynamin is critical for efficient viral entry.

  2. A METHOD TO REMOVE ENVIRONMENTAL INHIBITORS PRIOR TO THE DETECTION OF WATERBORNE ENTERIC VIRUSES BY REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION

    EPA Science Inventory

    A method was developed to remove environmental inhibitors from sample concentrates prior to detection of human enteric viruses using the reverse transcription-polymerase chain reaction (RT-PCR).Environmental inhibitors, concentrated along with viruses during water sample processi...

  3. Isolation of an adenovirus and an adeno-associated virus from goat kids with enteritis.

    PubMed

    Olson, Erik J; Haskell, Scott R R; Frank, Rodney K; Lehmkuhl, Howard D; Hobbs, Lea Ann; Warg, Janet V; Landgraf, John G; Wünschmann, Arno

    2004-09-01

    A dairy goat operation in Minnesota experienced a sudden, markedly increased mortality among its neonatal goats. Approximately 60 of 130 kids (46%) died. The animals had diarrhea and dyspnea of 1-2 days duration before death. Necropsy of 4 goat kids revealed marked, acute, catarrhal enteritis and fibrinous pleuropneumonia. Mannheimia haemolytica was isolated from the lungs. Basophilic inclusion bodies filling the entire nucleus were present in enterocytes of the ileum of 3 goats. Adenoviral particles were detected in the feces by electron microscopy and adenovirus was subsequently isolated from the intestinal content together with a parvo-like virus (dependovirus). Morphology, physicochemical characteristics, and neutralization tests indicated that the adenovirus resembled ovine adenovirus-2 (OAdV-2). However, the PstI restriction endonuclease pattern produced by the goat adenovirus was distinct from that of OAdV-2. This is the first report of enteritis in goats with an adenovirus antigenically related to OAdV-2 and with a parvo-like dependovirus.

  4. Live Attenuated Vaccine Based on Duck Enteritis Virus against Duck Hepatitis A Virus Types 1 and 3

    PubMed Central

    Zou, Zhong; Ma, Ji; Huang, Kun; Chen, Huanchun; Liu, Ziduo; Jin, Meilin

    2016-01-01

    As causative agents of duck viral hepatitis, duck hepatitis A virus type 1 (DHAV-1) and type 3 (DHAV-3) causes significant economic losses in the duck industry. However, a licensed commercial vaccine that simultaneously controls both pathogens is currently unavailable. Here, we generated duck enteritis virus recombinants (rC-KCE-2VP1) containing both VP1 from DHAV-1 (VP1/DHAV-1) and VP1 from DHAV-3 (VP1/DHAV-3) between UL27 and UL26. A self-cleaving 2A-element of FMDV was inserted between the two different types of VP1, allowing production of both proteins from a single open reading frame. Immunofluorescence and Western blot analysis results demonstrated that both VP1 proteins were robustly expressed in rC-KCE-2VP1-infected chicken embryo fibroblasts. Ducks that received a single dose of rC-KCE-2VP1 showed potent humoral and cellular immune responses and were completely protected against challenges of both pathogenic DHAV-1 and DHAV-3 strains. The protection was rapid, achieved as early as 3 days after vaccination. Moreover, viral replication was fully blocked in vaccinated ducks as early as 1 week post-vaccination. These results demonstrated, for the first time, that recombinant rC-KCE-2VP1 is potential fast-acting vaccine against DHAV-1 and DHAV-3. PMID:27777571

  5. Removal properties of human enteric viruses in a pilot-scale membrane bioreactor (MBR) process.

    PubMed

    Miura, Takayuki; Okabe, Satoshi; Nakahara, Yoshihito; Sano, Daisuke

    2015-05-15

    In order to evaluate removal properties of human enteric viruses from wastewater by a membrane bioreactor (MBR), influent, anoxic and oxic mixed liquor, and membrane effluent samples were collected in a pilot-scale anoxic-oxic MBR process for 16 months, and concentrations of enteroviruses, norovirus GII, and sapoviruses were determined by real-time PCR using murine norovirus as a process control. Mixed liquor samples were separated into liquid and solid phases by centrifugation, and viruses in the bulk solution and those associated with mixed liquor suspended solids (MLSS) were quantified. Enteroviruses, norovirus GII, and sapoviruses were detected in the influent throughout the sampling period (geometrical mean, 4.0, 3.1, and 4.4 log copies/mL, respectively). Enterovirus concentrations in the solid phase of mixed liquor were generally lower than those in the liquid phase, and the mean log reduction value between influent and anoxic mixed liquor was 0.40 log units. In contrast, norovirus GII and sapovirus concentrations in the solid phase were equal to or higher than those in the liquid phase, and higher log reduction values (1.3 and 1.1 log units, respectively) were observed between influent and anoxic mixed liquor. This suggested that enteroviruses were less associated with MLSS than norovirus GII and sapoviruses, resulting in lower enterovirus removal in the activated sludge process. Enteroviruses and norovirus GII were detected in the MBR effluent but sapoviruses were not in any effluent samples. When MLSS concentration was reduced to 50-60% of a normal operation level, passages of enteroviruses and norovirus GII through a PVDF microfiltration membrane were observed. Since rejection of viruses by the membrane was not related to trans-membrane pressure which was monitored as a parameter of membrane fouling, the results indicated that adsorption to MLSS plays an important role in virus removal by an MBR, and removal properties vary by viruses reflecting different

  6. High prevalence of enteric viruses in untreated individual drinking water sources and surface water in Slovenia.

    PubMed

    Steyer, Andrej; Torkar, Karmen Godič; Gutiérrez-Aguirre, Ion; Poljšak-Prijatelj, Mateja

    2011-09-01

    Waterborne infections have been shown to be important in outbreaks of gastroenteritis throughout the world. Although improved sanitary conditions are being progressively applied, fecal contaminations remain an emerging problem also in developed countries. The aim of our study was to investigate the prevalence of fecal contaminated water sources in Slovenia, including surface waters and groundwater sources throughout the country. In total, 152 water samples were investigated, of which 72 samples represents groundwater from individual wells, 17 samples from public collection supplies and 63 samples from surface stream waters. Two liters of untreated water samples were collected and concentrated by the adsorption/elution technique with positively charged filters followed by an additional ultracentrifugation step. Group A rotaviruses, noroviruses (genogroups I and II) and astroviruses were detected with real-time RT-PCR method in 69 (45.4%) out of 152 samples collected, of which 31/89 (34.8%) drinking water and 38/63 (60.3%) surface water samples were positive for at least one virus tested. In 30.3% of drinking water samples group A rotaviruses were detected (27/89), followed by noroviruses GI (2.2%; 2/89) and astroviruses (2.2%; 2/89). In drinking groundwater samples group A rotaviruses were detected in 27 out of 72 tested samples (37.5%), genogroup I noroviruses in two (2.8%), and human astroviruses in one (1.4%) samples. In surface water samples norovirus genogroup GII was the most frequently detected (41.3%; 26/63), followed by norovirus GI (33.3%; 21/63), human astrovirus (27.0%; 17/63) and group A rotavirus (17.5%; 11/63). Our study demonstrates relatively high percentage of groundwater contamination in Slovenia and, suggests that raw groundwater used as individual drinking water supply may constitute a possible source of enteric virus infections. In the future, testing for enteric viruses should be applied for drinking water sources in waterborne outbreaks.

  7. Three-Dimensional Structure of a Protozoal Double-Stranded RNA Virus That Infects the Enteric Pathogen Giardia lamblia

    PubMed Central

    Janssen, Mandy E. W.; Takagi, Yuko; Parent, Kristin N.; Cardone, Giovanni

    2014-01-01

    ABSTRACT Giardia lamblia virus (GLV) is a small, nonenveloped, nonsegmented double-stranded RNA (dsRNA) virus infecting Giardia lamblia, the most common protozoan pathogen of the human intestine and a major agent of waterborne diarrheal disease worldwide. GLV (genus Giardiavirus) is a member of family Totiviridae, along with several other groups of protozoal or fungal viruses, including Leishmania RNA viruses and Trichomonas vaginalis viruses. Interestingly, GLV is more closely related than other Totiviridae members to a group of recently discovered metazoan viruses that includes penaeid shrimp infectious myonecrosis virus (IMNV). Moreover, GLV is the only known protozoal dsRNA virus that can transmit efficiently by extracellular means, also like IMNV. In this study, we used transmission electron cryomicroscopy and icosahedral image reconstruction to examine the GLV virion at an estimated resolution of 6.0 Å. Its outermost diameter is 485 Å, making it the largest totivirus capsid analyzed to date. Structural comparisons of GLV and other totiviruses highlighted a related “T=2” capsid organization and a conserved helix-rich fold in the capsid subunits. In agreement with its unique capacity as a protozoal dsRNA virus to survive and transmit through extracellular environments, GLV was found to be more thermoresistant than Trichomonas vaginalis virus 1, but no specific protein machinery to mediate cell entry, such as the fiber complexes in IMNV, could be localized. These and other structural and biochemical findings provide a basis for future work to dissect the cell entry mechanism of GLV into a “primitive” (early-branching) eukaryotic host and an important enteric pathogen of humans. IMPORTANCE Numerous pathogenic bacteria, including Corynebacterium diphtheriae, Salmonella enterica, and Vibrio cholerae, are infected with lysogenic bacteriophages that contribute significantly to bacterial virulence. In line with this phenomenon, several pathogenic protozoa

  8. NDV HN gene C-terminal extension is not the determinant of the enteric tropism but influences the virus virulence

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many asymptomatic enteric Newcastle disease virus (NDV) strains contain a larger hemagglutinin-neuraminidase (HN) protein (616 amino acids, aa) than that (571 aa) of virulent respirotropic NDV strains. Therefore, it has been suspected that the 45 aa extension at the C-terminus of HN influences the v...

  9. Characterization of preferential flow pathways in a siliciclastic aquifer system using human enteric viruses and groundwater geochemistry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Human enteric viruses have been recognized as an emerging groundwater contaminant and are found only in human waste. In urban environments the most likely source of human waste is from sanitary sewers. Determining the travel time for near-surface contaminants to reach deep public supply wells is i...

  10. Protective effects of recombinant glycoprotein D based prime boost approach against duck enteritis virus in mice model.

    PubMed

    Aravind, S; Kamble, Nitin Machindra; Gaikwad, Satish S; Shukla, Sanjeev Kumar; Saravanan, R; Dey, Sohini; Mohan, C Madhan

    2015-11-01

    Duck virus enteritis, also known as duck plague, is an acute herpes viral infection of ducks caused by duck enteritis virus (DEV). The method of repeated immunization with a live attenuated vaccine has been used for the prevention and control of duck enteritis virus (DEV). However, the incidence of the disease in vaccinated flocks and latency reactivation are the major constraints in the present vaccination programme. The immunogenicity and protective efficacy afforded by intramuscular inoculation of plasmid DNA encoding DEV glycoprotein D (pCDNA-gD) followed by DEV gD expressed in Saccharomyces cerevisia (rgD) was assessed in a murine model. Compared with mice inoculated with DNA (pCDNA-gD) or protein (rgD) only, mice inoculated with the combination of gD DNA and protein had enhanced ELISA antibody titers to DEV and had accelerated clearance of virus following challenge infection. Furthermore, the highest levels of lymphocyte proliferation response, IL-4, IL-12 and IFN-γ production were induced following priming with the DNA vaccine and boosting with the rgD protein. For instance, the specially designed recombinant DEV vector vaccine would be the best choice to use in ducks. It offers an excellent solution to the low vaccination coverage rate in ducks. We expect that the application of this novel vaccine in the near future will greatly decrease the virus load in the environment and reduce outbreaks of DEV in ducks.

  11. Detection of Bacterial Indicators and Human and Bovine Enteric Viruses in Surface Water and Groundwater Sources Potentially Impacted by Animal and Human Wastes in Lower Yakima Valley, Washington▿

    PubMed Central

    Gibson, Kristen E.; Schwab, Kellogg J.

    2011-01-01

    Tangential flow ultrafiltration (UF) was used to concentrate and recover bacterial indicators and enteric viruses from 100 liters of groundwater (GW; n = 10) and surface water (SW; n = 11) samples collected in Lower Yakima Valley, WA. Human and bovine enteric viruses were analyzed in SW and GW concentrates by real-time PCR by using integrated inhibition detection. PMID:21075875

  12. Use of propidium monoazide in reverse transcriptase PCR to distinguish between infectious and noninfectious enteric viruses in water samples.

    PubMed

    Parshionikar, Sandhya; Laseke, Ian; Fout, G Shay

    2010-07-01

    Human enteric viruses can be present in untreated and inadequately treated drinking water. Molecular methods, such as the reverse transcriptase PCR (RT-PCR), can detect viral genomes in a few hours, but they cannot distinguish between infectious and noninfectious viruses. Since only infectious viruses are a public health concern, methods that not only are rapid but also provide information on the infectivity of viruses are of interest. The intercalating dye propidium monoazide (PMA) has been used for distinguishing between viable and nonviable bacteria with DNA genomes, but it has not been used to distinguish between infectious and noninfectious enteric viruses with RNA genomes. In this study, PMA in conjunction with RT-PCR (PMA-RT-PCR) was used to determine the infectivity of enteric RNA viruses in water. Coxsackievirus, poliovirus, echovirus, and Norwalk virus were rendered noninfectious or inactivated by treatment with heat (72 degrees C, 37 degrees C, and 19 degrees C) or hypochlorite. Infectious or native and noninfectious or inactivated viruses were treated with PMA. This was followed by RNA extraction and RT-PCR or quantitative RT-PCR (qRT-PCR) analysis. The PMA-RT-PCR results indicated that PMA treatment did not interfere with detection of infectious or native viruses but prevented detection of noninfectious or inactivated viruses that were rendered noninfectious or inactivated by treatment at 72 degrees C and 37 degrees C and by hypochlorite treatment. However, PMA-RT-PCR was unable to prevent detection of enteroviruses that were rendered noninfectious by treatment at 19 degrees C. After PMA treatment poliovirus that was rendered noninfectious by treatment at 37 degrees C was undetectable by qRT-PCR, but PMA treatment did not affect detection of Norwalk virus. PMA-RT-PCR was also shown to be effective for detecting infectious poliovirus in the presence of noninfectious virus and in an environmental matrix. We concluded that PMA can be used to differentiate

  13. Expression and distribution of the duck enteritis virus UL51 protein in experimentally infected ducks.

    PubMed

    Shen, Chanjuan; Cheng, Anchun; Wang, Mingshu; Xu, Chao; Jia, Renyong; Chen, Xiaoyue; Zhu, Dekang; Luo, Qihui; Cui, Hengmin; Zhou, Yi; Wang, Yin; Xu, Zhiwen; Chen, Zhengli; Wang, Xiaoyu

    2010-06-01

    To determine the expression and distribution of tegument proteins encoded by duck enteritis virus (DEV) UL51 gene in tissues of experimentally infected ducks, for the first time, an immunoperoxidase staining method to detect UL51 protein (UL51p) in paraffin-embedded tissues is reported. A rabbit anti-UL51 polyclonal serum, raised against a recombinant 6-His-UL51 fusion protein expressed in Escherichia coli, was prepared, purified, and used as primary antibodies. Fifty-eight 30-day-old DEV-free ducks were intramuscularly inoculated with the pathogenic DEV CHv strain as infection group, and two ducks were selected as preinfection group. The tissues were collected at sequential time points between 2 and 480 hr postinoculation (PI) and prepared for immunoperoxidase staining. DEV UL51p was first found in the spleen and liver at 8 hr PI; in the bursa of Fabricius and thymus at 12 hr PI; in the Harders glands, esophagus, small intestine (including the duodenum, jejunum, and ileum), and large intestine (including the caecum and rectum) at 24 hr PI; in the glandularis ventriculus at 48 hr PI; and in the pancreas, cerebrum, kidney, lung, and myocardium at 72 hr PI. Throughout the infection process, the UL51p was not seen in the muscle. Furthermore, the intensity of positive staining of DEV UL51p antigen in various tissues increased sharply from 8 to 96 hr PI, peaked during 120-144 hr PI, and then decreased steadily from 216 to 480 hr PI, suggesting that the expressional levels of DEV UL51p in systemic organs have a close correlation with the progression of duck virus enteritis (DVE) disease. A number of DEV UL51p was distributed in the bursa of Fabricius, thymus, spleen, liver, esophagus, small intestine, and large intestine of DEV-infected ducks, whereas less DEV UL51p was distributed in the Harders glands, glandularis ventriculus, cerebrum, kidney, lung, pancreas, and myocardium of DEV-infected ducks. Moreover, DEV UL51p can be expressed in the cytoplasm of various types

  14. Quantification of enteric viruses, pathogen indicators, and Salmonella bacteria in class B anaerobically digested biosolids by culture and molecular methods.

    PubMed

    Wong, Kelvin; Onan, Brandon M; Xagoraraki, Irene

    2010-10-01

    The most common class B biosolids in the United States are generated by mesophilic anaerobic digestion (MAD), and MAD biosolids have been used for land application. However, the pathogen levels in MAD biosolids are still unclear, especially with respect to enteric viruses. In this study, we determined the occurrence and the quantitative levels of enteric viruses and indicators in 12 MAD biosolid samples and of Salmonella enterica in 6 MAD biosolid samples. Three dewatered biosolid samples were also included in this study for purposes of comparison. Human adenoviruses (HAdV) had the highest gene levels and were detected more frequently than other enteric viruses. The gene levels of noroviruses (NV) reported were comparable to those of enteroviruses (EV) and human polyomaviruses (HPyV). The occurrence percentages of HAdV, HAdV species F, EV, NV GI, NV GII, and HPyV in MAD samples were 83, 83, 42, 50, 75, and 58%, respectively. No hepatitis A virus was detected. Infectious HAdV was detected more frequently than infectious EV, and all infectious HAdV were detected when samples were propagated in A549 cells. Based on most-probable-number (MPN) analysis, A549 cells were more susceptible to biosolid-associated viruses than BGM cells. All indicator levels in MAD biosolids were approximately 10(4) MPN or PFU per gram (dry), and the dewatered biosolids had significantly higher indicator levels than the MAD biosolids. Only two MAD samples tested positive for Salmonella enterica, where the concentration was below 1.0 MPN/4 g. This study provides a broad comparison of the prevalence of different enteric viruses in MAD biosolids and reports the first detection of noroviruses in class B biosolids. The observed high quantitative and infectivity levels of adenoviruses in MAD biosolids indicate that adenovirus is a good indicator for the evaluation of sludge treatment efficiency.

  15. Enteric viruses in Brazilian turkey flocks: single and multiple virus infection frequency according to age and clinical signs of intestinal disease.

    PubMed

    Moura-Alvarez, J; Chacon, J V; Scanavini, L S; Nuñez, L F N; Astolfi-Ferreira, C S; Jones, R C; Piantino Ferreira, A J

    2013-04-01

    Poult enteritis complex has been associated with enteritis and reduction in growth rates in commercial turkeys worldwide. Intestinal samples from 76 turkey flocks from different Brazilian states affected or not with intestinal disorders were evaluated for the presence of adenovirus groups 1 and 2 (TAV), astrovirus types 1 and 2 (TAstV-1 and TAstV-2), turkey coronavirus (TCoV), reovirus, rotavirus, and avian nephritis virus (ANV) using PCR. The percentage of positive samples was categorized according to the geographic origin, age of the flocks, and presence of clinical signs of intestinal disease. The percentage of samples that were positive for at least one virus was 93.4%, whereas the percentage of samples that were positive for more than one virus was 69.7%. An average of 3.20 viruses per sample was detected in turkeys in the growing phase of the production cycle (1 to 4 wk of age). The TAstV-1 and TCoV were the most frequently observed viruses in growing phase turkeys and occurred simultaneously in 85% of these samples. In turkeys in the finishing phase of development (5 to 18 wk), a lower average number of viruses was observed (2.41), and the most frequent viruses isolated in these turkeys were TAstV-1 (57.1%) and rotavirus (51.8%). Overall, every virus was detected more frequently in growing phase turkeys than in finishing phase turkeys with the exception of TAV. Samples from flocks exhibiting clinical signs of intestinal disease showed a higher rate of positivity, and TAstV-1, TAstV-2, and TCoV were the most frequently occurring viruses in this cohort. Birds without clinical signs most frequently harbored TAstV-1 and rotavirus. Future studies should focus on the description and elucidation of the role of each virus, as well as the pathogenic and immunological implications of the different combinations of viruses in turkeys.

  16. DETECTION BY PCR OF HUMAN ENTERIC VIRUSES CONCENTRATED FROM LARGE VOLUMES OF WATER

    EPA Science Inventory

    Viruses are recovered and concentrated from water by passage through a positively charged cartridge filter. Following virus elution from the cartridge filter with beef extract and concentration of the beef extract solution, viruses are usually assayed by cell culture. However...

  17. MHC class I molecules are enriched in caveolae but do not enter with simian virus 40.

    PubMed

    Anderson, H A; Chen, Y; Norkin, L C

    1998-06-01

    Simian virus 40 (SV40) binds to MHC class I molecules anywhere on the cell surface and then enters through caveolae. The fate of class I molecules after SV40 binding is not known. Sensitivity of 125I-surface-labelled class I molecules to papain cleavage was used to distinguish internalized class I molecules from class I molecules remaining at the cell surface. Whereas the caveolae-enriched membrane microdomain was found to also be enriched for class I molecules, no internalized papain-resistant 125I-surface-labelled class I molecules could be detected at any time in either control cells or in cells preadsorbed with saturating amounts of SV40. Instead, 125I-surface-labelled class I molecules, as well as preadsorbed 125I-labelled anti-class I antibodies, accumulated in the medium, coincident with the turnover of class I molecules at the cell surface. The class I heavy chains that accumulated in the medium were truncated and their release was specifically prevented by the metalloprotease inhibitor 1,10-phenanthroline. Thus, whereas class I molecules mediate SV40 binding, they do not appear to mediate SV40 entry.

  18. Sunlight inactivation of somatic coliphage in the presence of natural organic matter.

    PubMed

    Sun, Chen-Xi; Kitajima, Masaaki; Gin, Karina Yew-Hoong

    2016-01-15

    Long wavelengths of sunlight spectrum (UVA and visible light), as well as natural organic matter (NOM) are important environmental factors affecting survival of viruses in aquatic environment through direct and indirect inactivation. In order to understand the virus inactivation kinetics under such conditions, this study investigated the effects of Suwannee River natural organic matter (NOM) on the inactivation of a somatic coliphage, phiX174, by UVA and visible light. Experiments were carried out to examine the virucidal effects of UVA/visible light, assess the influence of SRNOM at different concentrations, and identify the effective ROS in virus inactivation. The results from this study showed that the presence of NOM could either enhance virus inactivation or reduce virus inactivation depending on the concentration, where the inactivation rate followed a parabolic relationship against NOM concentration. The results indicated that moderate levels of NOM (11 ppm) had the strongest antiviral activity, while very low or very high NOM concentrations prolonged virus survival. The results also showed that OH▪ was the primary ROS in causing phiX174 (ssDNA virus) inactivation, unlike previous findings where (1)O2 was the primary ROS causing MS2 (ssRNA virus) inactivation. The phiX174 inactivation by OH∙ could be described as k=3.7 ✕ 10(13)[OH∙]+1.404 (R(2)=0.8527).

  19. Preparation of MS2 Phage-Like Particles and Their Use As Potential Process Control Viruses for Detection and Quantification of Enteric RNA Viruses in Different Matrices

    PubMed Central

    Mikel, Pavel; Vasickova, Petra; Tesarik, Radek; Malenovska, Hana; Kulich, Pavel; Vesely, Tomas; Kralik, Petr

    2016-01-01

    The detection and quantification of enteric RNA viruses is based on isolation of viral RNA from the sample followed by quantitative reverse transcription polymerase chain reaction (RT-qPCR). To control the whole process of analysis and in order to guarantee the validity and reliability of results, process control viruses (PCV) are used. The present article describes the process of preparation and use of such PCV– MS2 phage-like particles (MS2 PLP) – in RT-qPCR detection and quantification of enteric RNA viruses. The MS2 PLP were derived from bacteriophage MS2 carrying a unique and specific de novo-constructed RNA target sequence originating from the DNA of two extinct species. The amount of prepared MS2 particles was quantified using four independent methods – UV spectrophotometry, fluorimetry, transmission electron microscopy and a specifically developed duplex RT-qPCR. To evaluate the usefulness of MS2 PLP in routine diagnostics different matrices known to harbor enteric RNA viruses (swab samples, liver tissue, serum, feces, and vegetables) were artificially contaminated with specific amounts of MS2 PLP. The extraction efficiencies were calculated for each individual matrix. The prepared particles fulfill all requirements for PCV – they are very stable, non-infectious, and are genetically distinct from the target RNA viruses. Due to these properties they represent a good morphological and physiochemical model. The use of MS2 PLP as a PCV in detection and quantification of enteric RNA viruses was evaluated in different types of matrices. PMID:28133456

  20. Detection of human enteric viruses in oysters by in vivo and in vitro amplification of nucleic acids.

    PubMed Central

    Chung, H; Jaykus, L A; Sobsey, M D

    1996-01-01

    This study describes the detection of enteroviruses and hepatitis A virus in 31 naturally contaminated oyster specimens by nucleic acid amplification and oligonucleotide probing. Viruses were extracted by adsorption-elution-precipitation from 50-g oyster samples harvested from an area receiving sewage effluent discharge. Ninety percent of each extract was inoculated into primate kidney cell cultures for virus isolation and infectivity assay. Viruses in the remaining 10% of oyster extract that was not inoculated into cell cultures were further purified and concentrated by a procedure involving Freon extraction, polyethylene glycol precipitation, and Pro-Cipitate precipitation. After 3 to 4 weeks of incubation, RNA was extracted from inoculated cultures that were negative for cytopathic effects (CPE). These RNA extracts and the RNA from virions purified and concentrated directly from oyster extracts were subjected to reverse transcriptase PCR (RT-PCR) with primer pairs for human enteroviruses and hepatitis A virus. The resulting amplicons were confirmed by internal oligonucleotide probe hybridization. For the portions of oyster sample extracts inoculated into cell cultures, 12 (39%) were positive for human enteroviruses by CPE and 6 (19%) were positive by RT-PCR and oligoprobing of RNA extracts from CPE-negative cell cultures. For the remaining sample portions tested by direct RT-PCR and oligoprobing after further concentration, five (about 16%) were confirmed to be positive for human enteroviruses. Hepatitis A virus was also detected in RNA extracts of two CPE-positive samples by RT-PCR and oligoprobing. Combining the data from all three methods, enteric viruses were detected in 18 of 31 (58%) samples. Detection by nucleic acid methods increased the number of positive samples by 50% over detection by CPE in cell culture. Hence, nucleic acid amplification methods increase the detection of noncytopathic human enteric viruses in oysters. PMID:8837433

  1. Effect of distance from the polluting focus on relative concentrations of Bacteroides fragilis phages and coliphages in mussels.

    PubMed Central

    Lucena, F; Lasobras, J; McIntosh, D; Forcadell, M; Jofre, J

    1994-01-01

    Concentrations of fecal bacteria, somatic and F-specific coliphages, and phages infecting Bacteroides fragilis in naturally occurring black mussels (Mytilus edulis) were determined. Mussels were collected over a 7-month period at four sampling sites with different levels of fecal pollution. Concentrations of both fecal bacteria and bacteriophages in mussel meat paralleled the concentration of fecal bacteria in the overlying waters. Mussels bioaccumulated efficiently, although with different efficiencies, all of the microorganisms studied. Ratios comparing the levels of microorganisms in mussels were determined. These ratios changed in mussels collected at the different sites. They suggest that bacteriophages infecting B. fragilis and somatic coliphages have the lowest decay rates among the microorganisms studied, with the exception of Clostridium perfringens. On the contrary, concentrations of F-specific coliphages showed a greater rate of decay than the other bacteriophages at sites more distant from the focus of contamination. Additionally, levels of enteroviruses were studied in a number of samples, and in these samples, the B. fragilis bacteriophages clearly outnumbered the enteroviruses. The results of this study indicate that, under the environmental conditions studied, the fate of phages infecting B. fragilis released into the marine environment resembles that of human viruses more than any other microorganism examined. Images PMID:8074509

  2. The pathogenesis of duck virus enteritis in experimentally infected ducks: a quantitative time-course study using TaqMan polymerase chain reaction.

    PubMed

    Xuefeng, Qi; Xiaoyan, Yang; Anchun, Cheng; Mingshu, Wang; Dekang, Zhu; Renyong, Jia

    2008-06-01

    Duck virus enteritis is an acute and contagious herpesvirus infection of duck, geese and swans with high morbidity and mortality. The kinetics of viral DNA loads and immunohistochemical localization of virulent duck enteritis virus, as well as histopathological examination in various tissues of ducks following oral infection, were investigated. The time course for the appearance of viral antigen and tissue lesions in various tissues was coincident with the levels of duck enteritis virus at the various sites, suggesting that the levels of duck enteritis virus in systemic organs have a close correlation with the progression of disease. The abundance of target epithelial and lymphoid cells may contribute to the high levels of virus infection and replication in lymphoid and intestinal tissues.

  3. Presence of pathogenic enteric viruses in illegally imported meat and meat products to EU by international air travelers.

    PubMed

    Rodríguez-Lázaro, David; Diez-Valcarce, Marta; Montes-Briones, Rebeca; Gallego, David; Hernández, Marta; Rovira, Jordi

    2015-09-16

    One hundred twenty two meat samples confiscated from passengers on flights from non-European countries at the International Airport of Bilbao (Spain) were tested for the presence of the main foodborne viral pathogens (human noroviruses genogroups I and II, hepatitis A and E viruses) during 2012 and 2013. A sample process control virus, murine norovirus, was used to evaluate the correct performance of the method. Overall, 67 samples were positive for at least one enteric viruses, 65 being positive for hepatitis E virus (53.3%), 3 for human norovirus genogroup I (2.5%) and 1 for human norovirus genogroup II (0.8%), whereas hepatitis A virus was not detected in any sample. The type of positive meat samples was diverse, but mainly was pork meat products (64.2%). The geographical origin of the positive samples was wide and diverse; samples from 15 out 19 countries tested were positive for at least one virus. However, the estimated virus load was low, ranging from 55 to 9.0 × 10(4) PDU per gram of product. The results obtained showed the potential introduction of viral agents in travelers' luggage, which constitute a neglected route of introduction and transmission.

  4. Quantifying viruses and bacteria in wastewater—Results, interpretation methods, and quality control

    USGS Publications Warehouse

    Francy, Donna S.; Stelzer, Erin A.; Bushon, Rebecca N.; Brady, Amie M.G.; Mailot, Brian E.; Spencer, Susan K.; Borchardt, Mark A.; Elber, Ashley G.; Riddell, Kimberly R.; Gellner, Terry M.

    2011-01-01

    Membrane bioreactors (MBR), used for wastewater treatment in Ohio and elsewhere in the United States, have pore sizes small enough to theoretically reduce concentrations of protozoa and bacteria, but not viruses. Sampling for viruses in wastewater is seldom done and not required. Instead, the bacterial indicators Escherichia coli (E. coli) and fecal coliforms are the required microbial measures of effluents for wastewater-discharge permits. Information is needed on the effectiveness of MBRs in removing human enteric viruses from wastewaters, particularly as compared to conventional wastewater treatment before and after disinfection. A total of 73 regular and 28 quality-control (QC) samples were collected at three MBR and two conventional wastewater plants in Ohio during 23 regular and 3 QC sampling trips in 2008-10. Samples were collected at various stages in the treatment processes and analyzed for bacterial indicators E. coli, fecal coliforms, and enterococci by membrane filtration; somatic and F-specific coliphage by the single agar layer (SAL) method; adenovirus, enterovirus, norovirus GI and GII, rotavirus, and hepatitis A virus by molecular methods; and viruses by cell culture. While addressing the main objective of the study-comparing removal of viruses and bacterial indicators in MBR and conventional plants-it was realized that work was needed to identify data analysis and quantification methods for interpreting enteric virus and QC data. Therefore, methods for quantifying viruses, qualifying results, and applying QC data to interpretations are described in this report. During each regular sampling trip, samples were collected (1) before conventional or MBR treatment (post-preliminary), (2) after secondary or MBR treatment (post-secondary or post-MBR), (3) after tertiary treatment (one conventional plant only), and (4) after disinfection (post-disinfection). Glass-wool fiber filtration was used to concentrate enteric viruses from large volumes, and small

  5. Assessment and Evaluation of an Integrated Hybrid Anaerobic-Aerobic Sewage Treatment System for the Removal of Enteric Viruses.

    PubMed

    El-Senousy, Waled Morsy; Abou-Elela, Sohair Imam

    2017-02-14

    The capability of a cost-effective and a small size decentralized pilot wastewater treatment plant (WWTP) to remove enteric viruses such as rotavirus, norovirus genogroup I (GGI), norovirus genogroup II (GGII), Hepatitis E virus (HEV), and adenovirus was studied. This pilot plant is an integrated hybrid anaerobic/aerobic setup which consisted of anaerobic sludge blanket (UASB), biological aerated filter (BAF), and inclined plate settler (IPS). Both the UASB and BAF are packed with a non-woven polyester fabric (NWPF). Results indicated that the overall log10 reductions of enteric viruses' genome copies through the whole system were 3.1 ± 1, 3.3 ± 0.5, and 2.6 ± 0.9 log10 for rotavirus, norovirus GGI, and adenovirus, respectively. Reduction efficiency for both norovirus GGII and HEV after the different treatment steps could not be calculated because there were no significant numbers of positive samples for both viruses. The overall reduction of rotavirus infectious units through the whole system was 2.2 ± 0.8 log10 reduction which is very close to the overall log10 reduction of adenovirus infectious units through the whole system which was 2.1 ± 0.8 log10 reduction. There was no considerable difference in the removal efficiency for different rotavirus G and P types. Adenovirus 41 was the only type detected in the all positive samples. Although the pilot WWTP investigated is cost effective, has a small footprint, does not need a long distance network pipes, and easy to operate, its efficiency to remove enteric viruses is comparable with the conventional centralized WWTPs.

  6. COLIPHAGES AS POTENTIAL VIRAL INDICATORS OF FECAL POLLUTION

    EPA Science Inventory

    Friedman, Stephanie D. In press. Coliphages as Potential Viral Indicators of Fecal Pollution (Abstract). To be presented at the SWS/GERS Fall Joint Society Meeting: Communication and Collaboration: Coastal Systems of the Gulf of Mexico and Southeastern United States, 6-9 October ...

  7. QUALITY ASSURANCE FOR METHODS TO DETECT HUMAN ENTERIC VIRUSES IN DRINKING WATER

    EPA Science Inventory

    Surface or groundwaters impacted by untreated or inadequately treated domestic wastes may contain human pathogenic viruses that cause hepatitis, gastroenteritis, meningitis, encephalitis, myocarditis, diabetes, conjunctivitis and temporary or permanent paralysis. These viruses c...

  8. Human enteric viruses in groundwater indicate offshore transport of human sewage to coral reefs of the Upper Florida Keys

    USGS Publications Warehouse

    Futch, J. Carrie; Griffin, Dale W.; Lipp, Erin K.

    2010-01-01

    To address the issue of human sewage reaching corals along the main reef of the Florida Keys, samples were collected from surface water, groundwater and coral [surface mucopolysaccharide layers (SML)] along a 10 km transect near Key Largo, FL. Samples were collected semi-annually between July 2003 and September 2005 and processed for faecal indicator bacteria (faecal coliform bacteria, enterococci and Clostridium perfringens) and human-specific enteric viruses (enterovirus RNA and adenovirus DNA) by (RT)-nested polymerase chain reaction. Faecal indicator bacteria concentrations were generally higher nearshore and in the coral SML. Enteric viruses were evenly distributed across the transect stations. Adenoviruses were detected in 37 of 75 samples collected (49.3%) whereas enteroviruses were only found in 8 of 75 samples (10.7%). Both viruses were detected twice as frequently in coral compared with surface water or groundwater. Offshore, viruses were most likely to be found in groundwater, especially during the wet summer season. These data suggest that polluted groundwater may be moving to the outer reef environment in the Florida Keys.

  9. Detection of multiple enteric virus strains within a foodborne outbreak of gastroenteritis: an indication of the source of contamination.

    PubMed Central

    Gallimore, C. I.; Pipkin, C.; Shrimpton, H.; Green, A. D.; Pickford, Y.; McCartney, C.; Sutherland, G.; Brown, D. W. G.; Gray, J. J.

    2005-01-01

    An outbreak of acute gastroenteritis of suspected viral aetiology occurred in April 2003 in the British Royal Fleet Auxiliary ship (RFA) Argus deployed in the Northern Arabian Gulf. There were 37 cases amongst a crew of 400 personnel. Of 13 samples examined from cases amongst the crew, six enteric viruses were detected by reverse transcriptase polymerase chain reaction (RT-PCR). Five different viruses were identified including, three norovirus genotypes, a sapovirus and a rotavirus. No multiple infections were detected. A common food source was implicated in the outbreak and epidemiological analysis showed a statistically significant association with salad as the source of the outbreak, with a relative risk of 3.41 (95% confidence interval of 1.7-6.81) of eating salad on a particular date prior to the onset of symptoms. Faecal contamination of the salad at source was the most probable explanation for the diversity of viruses detected and characterized. PMID:15724709

  10. One-year Surveillance of Human Enteric Viruses in Raw and Treated Wastewaters, Downstream River Waters, and Drinking Waters.

    PubMed

    Iaconelli, M; Muscillo, M; Della Libera, S; Fratini, M; Meucci, L; De Ceglia, M; Giacosa, D; La Rosa, G

    2017-03-01

    Human enteric viruses are a major cause of waterborne diseases, and can be transmitted by contaminated water of all kinds, including drinking and recreational water. The objectives of the present study were to assess the occurrence of enteric viruses (enterovirus, norovirus, adenovirus, hepatitis A and E virus) in raw and treated wastewaters, in rivers receiving wastewater discharges, and in drinking waters. Wastewater treatment plants' (WWTP) pathogen removal efficiencies by adenovirus quantitative real-time PCR and the presence of infectious enterovirus, by cell culture assays, in treated wastewaters and in surface waters were also evaluated. A total of 90 water samples were collected: raw and treated wastewaters (treated effluents and ultrafiltered water reused for industrial purposes), water from two rivers receiving treated discharges, and drinking water. Nested PCR assays were used for the identification of viral DNA/RNA, followed by direct amplicon sequencing. All raw sewage samples (21/21), 61.9 % of treated wastewater samples (13/21), and 25 % of ultrafiltered water samples (3/12) were contaminated with at least one viral family. Multiple virus families and genera were frequently detected. Mean positive PCRs per sample decreased significantly from raw to treated sewage and to ultrafiltered waters. Moreover, quantitative adenovirus data showed a reduction in excess of 99 % in viral genome copies following wastewater treatment. In surface waters, 78.6 % (22/28) of samples tested positive for one or more viruses by molecular methods, but enterovirus-specific infectivity assays did not reveal infectious particles in these samples. All drinking water samples tested negative for all viruses, demonstrating the effectiveness of treatment in removing viral pathogens from drinking water. Integrated strategies to manage water from all sources are crucial to ensure water quality.

  11. The vaccine efficacy of recombinant duck enteritis virus expressing secreted E with or without PrM proteins of duck tembusu virus.

    PubMed

    Chen, Pucheng; Liu, Jinxiong; Jiang, Yongping; Zhao, Yuhui; Li, Qimeng; Wu, Li; He, Xijun; Chen, Hualan

    2014-09-15

    A newly emerged tembusu virus that causes egg-drop has been affecting ducks in China since 2010. Currently, no vaccine is available for this disease. A live attenuated duck enteritis virus (DEV; a herpesvirus) vaccine has been used routinely to control lethal DEV in ducks since the 1960s. Here, we constructed two recombinant DEVs by transfecting overlapping fosmid DNAs. One virus, rDEV-TE, expresses the truncated form of the envelope glycoprotein (TE) of duck tembusu virus (DTMUV), and the other virus, rDEV-PrM/TE, expresses both the TE and pre-membrane proteins (PrM). Animal study demonstrated that both recombinant viruses induced measurable anti-DTMUV neutralizing antibodies in ducks. After two doses of recombinant virus, rDEV-PrM/TE completely protected ducks from DTMUV challenge, whereas rDEV-TE only conferred partial protection. These results demonstrate that recombinant DEV expressing the TE and pre-membrane proteins is protective and can serve as a potential candidate vaccine to prevent DTMUV infection in ducks.

  12. Source and transport of human enteric viruses in deep municipal water supply wells

    USGS Publications Warehouse

    Bradbury, Kenneth R.; Borchardt, Mark A.; Gotkowitz, Madeline; Spencer, Susan K.; Zhu, Jun; Hunt, Randall J.

    2013-01-01

    Until recently, few water utilities or researchers were aware of possible virus presence in deep aquifers and wells. During 2008 and 2009 we collected a time series of virus samples from six deep municipal water-supply wells. The wells range in depth from approximately 220 to 300 m and draw water from a sandstone aquifer. Three of these wells draw water from beneath a regional aquitard, and three draw water from both above and below the aquitard. We also sampled a local lake and untreated sewage as potential virus sources. Viruses were detected up to 61% of the time in each well sampled, and many groundwater samples were positive for virus infectivity. Lake samples contained viruses over 75% of the time. Virus concentrations and serotypes observed varied markedly with time in all samples. Sewage samples were all extremely high in virus concentration. Virus serotypes detected in sewage and groundwater were temporally correlated, suggesting very rapid virus transport, on the order of weeks, from the source(s) to wells. Adenovirus and enterovirus levels in the wells were associated with precipitation events. The most likely source of the viruses in the wells was leakage of untreated sewage from sanitary sewer pipes.

  13. Source and transport of human enteric viruses in deep municipal water supply wells.

    PubMed

    Bradbury, Kenneth R; Borchardt, Mark A; Gotkowitz, Madeline; Spencer, Susan K; Zhu, Jun; Hunt, Randall J

    2013-05-07

    Until recently, few water utilities or researchers were aware of possible virus presence in deep aquifers and wells. During 2008 and 2009 we collected a time series of virus samples from six deep municipal water-supply wells. The wells range in depth from approximately 220 to 300 m and draw water from a sandstone aquifer. Three of these wells draw water from beneath a regional aquitard, and three draw water from both above and below the aquitard. We also sampled a local lake and untreated sewage as potential virus sources. Viruses were detected up to 61% of the time in each well sampled, and many groundwater samples were positive for virus infectivity. Lake samples contained viruses over 75% of the time. Virus concentrations and serotypes observed varied markedly with time in all samples. Sewage samples were all extremely high in virus concentration. Virus serotypes detected in sewage and groundwater were temporally correlated, suggesting very rapid virus transport, on the order of weeks, from the source(s) to wells. Adenovirus and enterovirus levels in the wells were associated with precipitation events. The most likely source of the viruses in the wells was leakage of untreated sewage from sanitary sewer pipes.

  14. Expression and characterization of UL16 gene from duck enteritis virus

    PubMed Central

    2011-01-01

    Background Previous studies have indicated that the UL16 protein and its homologs from herpesvirus were conserved and played similar roles in viral DNA packaging, virion assembly, budding, and egress. However, there was no report on the UL16 gene product of duck enteritis virus (DEV). In this study, we analyzed the amino acid sequence of UL16 using bioinformatics tools and expressed in Escherichia coli Rosetta (DE3) induced by isopropy1-β-D-thiogalactopyranoside (IPTG). The recombinant protein was produced, purified using a Ni-NTA column and used to generate the polyclonal antibody against UL16. The intracellular distribution of the DEV UL16 product was carried out using indirect immunofluorescence assay. Results In our study, UL16 gene of DEV was composed of 1089 nucleotides, which encoded 362 amino acids. Multiple sequence alignment suggested that the UL16 gene was highly conserved in herpesvirus family. The UL16 gene was cloned into a pET prokaryotic expression vector and transformed into Escherichia coli Rossetta (DE3) induced by IPTG. A 60kDa fusion protein band corresponding to the predicted size was produced on the SDS-PAGE, purified using a Ni-NTA column. Anti-UL16 polyclonal sera was prepared by immunizing rabbits, and reacted with a band in the IPTG induced cell lysates with an apparent molecular mass of 60 kDa. In vivo expression of the UL16 protein in DEV infected duck embryo fibroblast cells (DEFs) was localized mostly around perinuclear cytoplasmic area and in cytosol using indirect immunofluorescence assay. Conclusions The UL16 gene of DEV was successfully cloned, expressed and detected in DEV infected DEFs for the first time. The UL16 protein localized mostly around perinuclear cytoplasmic area and in cytosol in DEV infected DEFs. DEV UL16 shared high similarity with UL16 family members, indicating that DEV UL16 many has similar function with its homologs. All these results may provide some insight for further research about full characterizations

  15. Metagenomic identification of novel enteric viruses in urban wild rats and genome characterization of a group A rotavirus

    PubMed Central

    Sachsenröder, Jana; Braun, Anne; Machnowska, Patrycja; Ng, Terry Fei Fan; Deng, Xutao; Guenther, Sebastian; Bernstein, Samuel; Ulrich, Rainer G.; Delwart, Eric

    2014-01-01

    Rats are known as reservoirs and vectors for several zoonotic pathogens. However, information on the viruses shed by urban wild rats that could pose a zoonotic risk to human health is scare. Here, intestinal contents from 20 wild Norway rats (Rattus norvegicus) collected in the city of Berlin, Germany, were subjected to metagenomic analysis of viral nucleic acids. The determined faecal viromes of rats consisted of a variety of known and unknown viruses, and were highly variable among the individuals. Members of the families Parvoviridae and Picobirnaviridae represented the most abundant species. Novel picornaviruses, bocaviruses, sapoviruses and stool-associated circular ssDNA viruses were identified, which showed only low sequence identity to known representatives of the corresponding taxa. In addition, noroviruses and rotaviruses were detected as potential zoonotic gastroenteritis viruses. However, partial-genome sequence analyses indicated that the norovirus was closely related to the recently identified rat norovirus and the rotavirus B was closely related to the rat rotavirus strain IDIR; both viruses clustered separately from respective human virus strains in phylogenetic trees. In contrast, the rotavirus A sequences showed high identity to human and animal strains. Analysis of the nearly complete genome of this virus revealed the known genotypes G3, P[3] and N2 for three of the genome segments, whereas the remaining eight genome segments represented the novel genotypes I20–R11–C11–M10–A22–T14–E18–H13. Our results indicated a high heterogeneity of enteric viruses present in urban wild rats; their ability to be transmitted to humans remains to be assessed in the future. PMID:25121550

  16. Metagenomic identification of novel enteric viruses in urban wild rats and genome characterization of a group A rotavirus.

    PubMed

    Sachsenröder, Jana; Braun, Anne; Machnowska, Patrycja; Ng, Terry Fei Fan; Deng, Xutao; Guenther, Sebastian; Bernstein, Samuel; Ulrich, Rainer G; Delwart, Eric; Johne, Reimar

    2014-12-01

    Rats are known as reservoirs and vectors for several zoonotic pathogens. However, information on the viruses shed by urban wild rats that could pose a zoonotic risk to human health is scare. Here, intestinal contents from 20 wild Norway rats (Rattus norvegicus) collected in the city of Berlin, Germany, were subjected to metagenomic analysis of viral nucleic acids. The determined faecal viromes of rats consisted of a variety of known and unknown viruses, and were highly variable among the individuals. Members of the families Parvoviridae and Picobirnaviridae represented the most abundant species. Novel picornaviruses, bocaviruses, sapoviruses and stool-associated circular ssDNA viruses were identified, which showed only low sequence identity to known representatives of the corresponding taxa. In addition, noroviruses and rotaviruses were detected as potential zoonotic gastroenteritis viruses. However, partial-genome sequence analyses indicated that the norovirus was closely related to the recently identified rat norovirus and the rotavirus B was closely related to the rat rotavirus strain IDIR; both viruses clustered separately from respective human virus strains in phylogenetic trees. In contrast, the rotavirus A sequences showed high identity to human and animal strains. Analysis of the nearly complete genome of this virus revealed the known genotypes G3, P[3] and N2 for three of the genome segments, whereas the remaining eight genome segments represented the novel genotypes I20-R11-C11-M10-A22-T14-E18-H13. Our results indicated a high heterogeneity of enteric viruses present in urban wild rats; their ability to be transmitted to humans remains to be assessed in the future.

  17. Reduction of Norwalk virus, poliovirus 1, and bacteriophage MS2 by ozone disinfection of water.

    PubMed

    Shin, Gwy-Am; Sobsey, Mark D

    2003-07-01

    Norwalk virus and other human caliciviruses (noroviruses) are major agents of gastroenteritis, and water is a major route of their transmission. In an effort to control Norwalk virus in drinking water, Norwalk virus reduction by bench-scale ozone disinfection was determined using quantitative reverse transcription (RT)-PCR for virus assays. Two other enteric viruses, poliovirus 1 and coliphage MS2, were included for comparison, and their reductions were assayed by infectivity assays as well as by RT-PCR. Virus reductions by ozone were determined using a dose of 0.37 mg of ozone/liter at pH 7 and 5 degrees C for up to 5 min. Based on two RT-PCR assays, the reductions of Norwalk virus were >3 log(10) within a contact time of 10 s, and these were similar to the reductions of the other two viruses determined by the same assay methods. Also, the virus reductions detected by RT-PCR assays were similar to those detected by infectivity assays, indicating that the RT-PCR assay is a reliable surrogate assay for both culturable and nonculturable viruses disinfected with ozone. Overall, the results of this study indicate that Norwalk virus as well as other enteric viruses can be reduced rapidly and extensively by ozone disinfection and that RT-PCR is a useful surrogate assay for both culturable and nonculturable viruses disinfected with ozone.

  18. Construction of a recombinant duck enteritis virus vaccine expressing hemagglutinin of H9N2 avian influenza virus and evaluation of its efficacy in ducks.

    PubMed

    Sun, Ying; Yang, Chenghuai; Li, Junping; Li, Ling; Cao, Minghui; Li, Qihong; Li, Huijiao

    2017-01-01

    H9 subtype avian influenza viruses (AIVs) remain a significant burden in the poultry industry and are considered to be one of the most likely causes of any new influenza pandemic in humans. As ducks play an important role in the maintenance of H9 viruses in nature, successful control of the spread of H9 AIVs in ducks will have significant beneficial effects on public health. Duck enteritis virus (DEV) may be a promising candidate viral vector for aquatic poultry vaccination. In this study, we constructed a recombinant DEV, rDEV-∆UL2-HA, inserting the hemagglutinin (HA) gene from duck-origin H9N2 AIV into the UL2 gene by homologous recombination. One-step growth analyses showed that the HA gene insertion had no effect on viral replication and suggested that the UL2 gene was nonessential for virus growth in vitro. In vivo tests further showed that the insertion of the HA gene in place of the UL2 gene did not affect the immunogenicity of the virus. Moreover, a single dose of 10(3) TCID50 of rDEV-∆UL2-HA induced solid protection against lethal DEV challenge and completely prevented H9N2 AIV viral shedding. To our knowledge, this is the first report of a DEV-vectored vaccine providing robust protection against both DEV and H9N2 AIV virus infections in ducks.

  19. Oral Phage Therapy of Acute Bacterial Diarrhea With Two Coliphage Preparations: A Randomized Trial in Children From Bangladesh

    PubMed Central

    Sarker, Shafiqul Alam; Sultana, Shamima; Reuteler, Gloria; Moine, Deborah; Descombes, Patrick; Charton, Florence; Bourdin, Gilles; McCallin, Shawna; Ngom-Bru, Catherine; Neville, Tara; Akter, Mahmuda; Huq, Sayeeda; Qadri, Firdausi; Talukdar, Kaisar; Kassam, Mohamed; Delley, Michèle; Loiseau, Chloe; Deng, Ying; El Aidy, Sahar; Berger, Bernard; Brüssow, Harald

    2016-01-01

    Background Antibiotic resistance is rising in important bacterial pathogens. Phage therapy (PT), the use of bacterial viruses infecting the pathogen in a species-specific way, is a potential alternative. Method T4-like coliphages or a commercial Russian coliphage product or placebo was orally given over 4 days to Bangladeshi children hospitalized with acute bacterial diarrhea. Safety of oral phage was assessed clinically and by functional tests; coliphage and Escherichia coli titers and enteropathogens were determined in stool and quantitative diarrhea parameters (stool output, stool frequency) were measured. Stool microbiota was studied by 16S rRNA gene sequencing; the genomes of four fecal Streptococcus isolates were sequenced. Findings No adverse events attributable to oral phage application were observed (primary safety outcome). Fecal coliphage was increased in treated over control children, but the titers did not show substantial intestinal phage replication (secondary microbiology outcome). 60% of the children suffered from a microbiologically proven E. coli diarrhea; the most frequent diagnosis was ETEC infections. Bacterial co-pathogens were also detected. Half of the patients contained phage-susceptible E. coli colonies in the stool. E. coli represented less than 5% of fecal bacteria. Stool ETEC titers showed only a short-lived peak and were otherwise close to the replication threshold determined for T4 phage in vitro. An interim analysis after the enrollment of 120 patients showed no amelioration in quantitative diarrhea parameter by PT over standard care (tertiary clinical outcome). Stool microbiota was characterized by an overgrowth with Streptococcus belonging to the Streptococcus gallolyticus and Streptococcus salivarius species groups, their abundance correlated with quantitative diarrhea outcome, but genome sequencing did not identify virulence genes. Interpretation Oral coliphages showed a safe gut transit in children, but failed to achieve

  20. Molecular epidemiology of enteric viruses in patients with acute gastroenteritis in Aichi prefecture, Japan, 2008/09-2013/14.

    PubMed

    Nakamura, Noriko; Kobayashi, Shinichi; Minagawa, Hiroko; Matsushita, Tadashi; Sugiura, Wataru; Iwatani, Yasumasa

    2016-07-01

    Acute gastroenteritis is a critical infectious disease that affects infants and young children throughout the world, including Japan. This retrospective study was conducted from September 2008 to August 2014 (six seasons: 2008/09-2013/14) to investigate the incidence of enteric viruses responsible for 1,871 cases of acute gastroenteritis in Aichi prefecture, Japan. Of the 1,871 cases, 1,100 enteric viruses were detected in 978 samples, of which strains from norovirus (NoV) genogroup II (60.9%) were the most commonly detected, followed by strains of rotavirus A (RVA) (23.2%), adenovirus (AdV) type 41 (8.2%), sapovirus (SaV) (3.6%), human astrovirus (HAstV) (2.8%), and NoV genogroup I (1.3%). Sequencing of the NoV genogroup II (GII) strains revealed that GII.4 was the most common genotype, although four different GII.4 variants were also identified. The most common G-genotype of RVA was G1 (63.9%), followed by G3 (27.1%), G2 (4.7%) and G9 (4.3%). Three genogroups of SaV strains were found: GI (80.0%), GII (15.0%), and GV (5.0%). HAstV strains were genotyped as HAstV-1 (80.6%), HAstV-8 (16.1%), and HAstV-3 (3.2%). These results show that NoV GII was the leading cause of sporadic acute viral gastroenteritis, although a variety of enteric viruses were detected during the six-season surveillance period.

  1. Occurrence of human enteric viruses at freshwater beaches during swimming season and its link to water inflow.

    PubMed

    Lee, Chang Soo; Lee, Cheonghoon; Marion, Jason; Wang, Qiuhong; Saif, Linda; Lee, Jiyoung

    2014-02-15

    Human enteric viruses are significant etiological agents for many recreational waterborne illnesses. The occurrence and density of human enteric viruses such as human adenovirus (HAdV), human enterovirus (HEnV), and human norovirus genogroups I/II (HNoV GI/GII) were investigated using quantitative real-time PCR (qPCR) at freshwater beaches along with monitoring fecal indicators and environmental parameters. During the 2009 swimming season, water samples were collected from three inland freshwater beaches in Ohio, USA. Of the total samples, 40% (26/65) and 17% (11/65) were positive for HAdV and HEnV respectively, but HNoV GI/GII were not detected. There was no significant association among the detected human enteric viruses (HAdV and HEnV) and fecal bacteria indicators (Escherichia coli and Bacteroides) by Spearman correlation and principal component analyses. Logistic regression analysis also revealed that the odds of finding HAdV or HEnV was not influenced by levels of fecal bacteria indicators. However, there was a 14-fold increase in the odds of HEnV detection for each 1-log increase in daily water inflow (m(3)/s) into freshwater beach reservoirs (adjusted odds ratio=14.2; 95% confidence interval=1.19-171). In summary, the viral occurrence at the freshwater beaches was not readily explained by the levels of fecal bacteria indicators, but appeared to be more related to water reservoir inflows. These results suggest that hydrological data must be considered in future epidemiology efforts aimed at characterizing beach water safety.

  2. Infected peripheral blood mononuclear cells transmit latent varicella zoster virus infection to the guinea pig enteric nervous system.

    PubMed

    Gan, Lin; Wang, Mingli; Chen, Jason J; Gershon, Michael D; Gershon, Anne A

    2014-10-01

    Latent wild-type (WT) and vaccine (vOka) varicella zoster virus (VZV) are found in the human enteric nervous system (ENS). VZV also infects guinea pig enteric neurons in vitro, establishes latency and can be reactivated. We therefore determined whether lymphocytes infected in vitro with VZV secrete infectious virions and can transfer infection in vivo to the ENS of recipient guinea pigs. T lymphocytes (CD3-immunoreactive) were preferentially infected following co-culture of guinea pig or human peripheral blood mononuclear cells with VZV-infected HELF. VZV proliferated in the infected T cells and expressed immediate early and late VZV genes. Electron microscopy confirmed that VZV-infected T cells produced encapsulated virions. Extracellular virus, however, was pleomorphic, suggesting degradation occurred prior to release, which was confirmed by the failure of VZV-infected T cells to secrete infectious virions. Intravenous injection of WT- or vOka-infected PBMCs, nevertheless, transmitted VZV to recipient animals (guinea pig > human lymphocytes). Two days post-inoculation, lung and liver, but not gut, contained DNA and transcripts encoding ORFs 4, 40, 66 and 67. Twenty-eight days after infection, gut contained DNA and transcripts encoding ORFs 4 and 66 but neither DNA nor transcripts could any longer be found in lung or liver. In situ hybridization revealed VZV DNA in enteric neurons, which also expressed ORF63p (but not ORF68p) immunoreactivity. Observations suggest that VZV infects T cells, which can transfer VZV to and establish latency in enteric neurons in vivo. Guinea pigs may be useful for studies of VZV pathogenesis in the ENS.

  3. Infected Peripheral Blood Mononuclear Cells Transmit Latent Varicella Zoster Virus Infection to the Guinea Pig Enteric Nervous System

    PubMed Central

    Gan, Lin; Wang, Mingli; Chen, Jason J.; Gershon, Michael D.; Gershon, Anne A.

    2014-01-01

    Latent wild-type (WT) and vaccine (vOka) varicella-zoster virus (VZV) are found in the human enteric nervous system (ENS). VZV also infects guinea pig enteric neurons in vitro, establishes latency and can be reactivated. We therefore determined whether lymphocytes infected in vitro with VZV secrete infectious virions and can transfer infection in vivo to the ENS of recipient guinea pigs. T lymphocytes (CD3-immunoreactive) were preferentially infected following co-culture of guinea pig or human peripheral blood mononuclear cells with VZV-infected HELF. VZV proliferated in the infected T cells and expressed immediate early and late VZV genes. Electron microscopy confirmed that VZV-infected T cells produced encapsulated virions. Extracellular virus, however, was pleomorphic, suggesting degradation occurred prior to release, which was confirmed by the failure of VZV-infected T cells to secrete infectious virions. Intravenous injection of WT- or vOka-infected PBMCs, nevertheless, transmitted VZV to recipient animals (guinea pig > human lymphocytes). Two days post-inoculation, lung and liver, but not gut, contained DNA and transcripts encoding ORFs 4, 40, 66 and 67. Twenty-eight days after infection, gut contained DNA and transcripts encoding ORFs 4 and 66 but neither DNA nor transcripts could any longer be found in lung or liver. In situ hybridization revealed VZV DNA in enteric neurons, which also expressed ORF63p (but not ORF68p) immunoreactivity. Observations suggest that VZV infects T cells, which can transfer VZV to and establish latency in enteric neurons in vivo. Guinea pigs may be useful for studies of VZV pathogenesis in the ENS. PMID:24965252

  4. Ozone disinfection dynamics of enteric viruses provide evidence that infectious titer reduction is triggered by alterations to viral colloidal properties.

    PubMed

    Vanden Bossche, G; Wustmann, U; Krietemeyer, S

    1994-11-01

    The inactivation dynamics of three enteric virus species (polio-, rota- and parvovirus) were analysed in different aqueous suspensions by using O3 under continuous flow conditions. A mathematical model for the reaction rate of infectious titer reduction was proposed, based on the thermodynamic principles of phase behaviour of colloids suspended in aqueous environments. Up to a certain threshold dosage of residual ozone (RO), and depending on the type of test virus and the ionic or organic load in the stock suspension, the logarithm of the reaction rate constant of viral inactivation rate was observed to vary in a rather sigmoidal manner with log RO concentration. Data from photon correlation spectroscopy, electron microscopy and tensiometric analysis suggested that below the threshold RO, the pattern of virus inactivation dynamics reflects the varying potential of different-sized viral particles (VPs) to adsorb to the cellular monolayer. There is strong evidence that oxidant-induced surface activity of organic matter causes redistribution of VP infectivity. This hypothesis was statistically corroborated inasmuch as experimental inactivation data proved to be satisfactorily fitted by a logistic equation. It was concluded that viral infection, and thus viral inactivation, is a complex process which is governed largely by the classical laws of colloidal behaviour. The latter is suggested to appreciably determine the capability of inoculated VPs to infect host cultures. This notion may especially be cause for concern when regulatory requirements for virus disinfection are being based on titration results from in vitro testing procedures.

  5. Data report. The fate of human enteric viruses in a natural sewage recycling system

    SciTech Connect

    Vaughn, J.M.; Landry, E.F.

    1980-09-01

    A two-year study was conducted to determine the virus-removing capacity of two man-made ecosystems designed for the treatment of raw domestic wastewater. The first treatment system consisted of two meadows followed by a marsh-pond unit (M/M/P). The second system contained individual marsh and pond units (M/P). All systems demonstrated moderate virus removal, with the marsh/pond system yielding the most consistent removal rates. Within this system, the greater potential for virus removal appeared to occur in the marsh unit. In addition to the production of system-oriented data, improved techniques for the concentration and enumeration of human viruses from sewage-polluted aquatic systems were developed.

  6. Post-epizootic surveys of waterfowl for duck plague (duck virus enteritis)

    USGS Publications Warehouse

    Brand, C.J.; Docherty, D.E.

    1988-01-01

    Surviving birds from nine duck plague outbreaks in urban and confined waterfowl were sampled for duck plague (DP) virus and DP antibody during 1979-86. Duck plague virus was found in combined oral and cloacal swabs of birds from three outbreaks, and DP-neutralizing antibody was demonstrated in some birds from all nine outbreaks. Greater prevalence of DP antibody and higher titers were found in survivors from confined populations than from free-flying urban populations. Free-flying waterfowl from within 52 km of four DP outbreak sites were also sampled; virus was not found in any birds, but DP antibody was found in urban waterfowl in the vicinity of an outbreak in Potterville, Michigan. No evidence of exposure to or shedding of DP virus in migratory waterfowl was found in two regions where DP appears enzootic in urban and confined waterfowl (Eastern Shore of Maryland and the vicinity of Sacramento, California).

  7. Chlorine inactivation of coliphage MS2 on strawberries by industrial-scale water washing units.

    PubMed

    Casteel, Michael J; Schmidt, Charles E; Sobsey, Mark D

    2009-06-01

    Fruits and vegetables (produce) intended for minimal processing are often rinsed or washed in water. Chlorine and other sanitizers are used during washing to inactivate produce spoilage microbes, but such procedures may also inactivate pathogens epidemiologically linked to produce, such as hepatitis A virus (HAV). However, no information exists on the efficacy of chlorinated wash water to inactivate HAV and other viruses on produce in actual practice, because of obvious safety concerns. In contrast, coliphage MS2 (a bacterial virus) is commonly used as a surrogate for some pathogenic viruses and may be safely used in field studies. In the present investigation, strawberries seeded with MS2 were passed through industrial-scale water washing units operated with or without added sodium hypochlorite. MS2 on strawberries was inactivated by 68%, 92% and 96% at free chlorine (FC) concentrations of < or = 2, 20 and 200 ppm in wash water, respectively. MS2 was detected in wash water containing < or = 2 ppm FC in one trial, but was not detected in water containing 20 or 200 ppm FC. The presence and absence of MS2 in wash water containing various levels of FC highlight the importance of controlling sanitizer levels to prevent viral cross contamination of strawberries.

  8. Detection of human enteric viruses in stream water with RT-PCR and cell culture.

    USGS Publications Warehouse

    Denis-Mize, K.; Fout, G.S.; Dahling, D.R.; Francy, D.S.

    2004-01-01

    A multiplex RT-PCR method was used to measure virus occurrence at five stream water sites that span a range of hydroclimatic, water-quality, and land-use characteristics. The performance of the molecular method was evaluated in comparison with traditional cell culture and Escherichia coli membrane filtration assays. The study incorporated multiple quality controls and included a control for virus recovery during the sampling procedure as well as controls to detect potentially false-negative and false-positive data. Poliovirus recovery ranged from 16 to 65% and was variable, even in samples collected within the same stream. All five sites were positive for viruses by both molecular and cell culture-based virus assays. Enteroviruses, reoviruses, rotaviruses, and hepatitis A viruses were detected, but the use of the quality controls proved critical for interpretation of the molecular data. All sites showed evidence of faecal contamination, and culturable viruses were detected in four samples that would have met the US Environmental Protection Agency's recommended E. coli guideline for safe recreational water.

  9. Human viruses and viral indicators in marine water at two recreational beaches in Southern California, USA.

    PubMed

    Love, David C; Rodriguez, Roberto A; Gibbons, Christopher D; Griffith, John F; Yu, Qilu; Stewart, Jill R; Sobsey, Mark D

    2014-03-01

    Waterborne enteric viruses may pose disease risks to bather health but occurrence of these viruses has been difficult to characterize at recreational beaches. The aim of this study was to evaluate water for human virus occurrence at two Southern California recreational beaches with a history of beach closures. Human enteric viruses (adenovirus and norovirus) and viral indicators (F+ and somatic coliphages) were measured in water samples over a 4-month period from Avalon Beach, Catalina Island (n = 324) and Doheny Beach, Orange County (n = 112). Human viruses were concentrated from 40 L samples and detected by nested reverse transcriptase polymerase chain reaction (PCR). Detection frequencies at Doheny Beach were 25.5% (adenovirus) and 22.3% (norovirus), and at Avalon Beach were 9.3% (adenovirus) and 0.7% (norovirus). Positive associations between adenoviruses and fecal coliforms were observed at Doheny (p = 0.02) and Avalon (p = 0.01) Beaches. Human viruses were present at both beaches at higher frequencies than previously detected in the region, suggesting that the virus detection methods presented here may better measure potential health risks to bathers. These virus recovery, concentration, and molecular detection methods are advancing practices so that analysis of enteric viruses can become more effective and routine for recreational water quality monitoring.

  10. A duck enteritis virus-vectored bivalent live vaccine provides fast and complete protection against H5N1 avian influenza virus infection in ducks.

    PubMed

    Liu, Jinxiong; Chen, Pucheng; Jiang, Yongping; Wu, Li; Zeng, Xianying; Tian, Guobin; Ge, Jinying; Kawaoka, Yoshihiro; Bu, Zhigao; Chen, Hualan

    2011-11-01

    Ducks play an important role in the maintenance of highly pathogenic H5N1 avian influenza viruses (AIVs) in nature, and the successful control of AIVs in ducks has important implications for the eradication of the disease in poultry and its prevention in humans. The inactivated influenza vaccine is expensive, labor-intensive, and usually needs 2 to 3 weeks to induce protective immunity in ducks. Live attenuated duck enteritis virus (DEV; a herpesvirus) vaccine is used routinely to control lethal DEV infections in many duck-producing areas. Here, we first established a system to generate the DEV vaccine strain by using the transfection of overlapping fosmid DNAs. Using this system, we constructed two recombinant viruses, rDEV-ul41HA and rDEV-us78HA, in which the hemagglutinin (HA) gene of the H5N1 virus A/duck/Anhui/1/06 was inserted and stably maintained within the ul41 gene or between the us7 and us8 genes of the DEV genome. Duck studies indicated that rDEV-us78HA had protective efficacy similar to that of the live DEV vaccine against lethal DEV challenge; importantly, a single dose of 10(6) PFU of rDEV-us78HA induced complete protection against a lethal H5N1 virus challenge in as little as 3 days postvaccination. The protective efficacy against both lethal DEV and H5N1 challenge provided by rDEV-ul41HA inoculation in ducks was slightly weaker than that provided by rDEV-us78HA. These results demonstrate, for the first time, that recombinant DEV is suitable for use as a bivalent live attenuated vaccine, providing rapid protection against both DEV and H5N1 virus infection in ducks.

  11. Presence, infectivity, and stability of enteric viruses in seawater: Relationship to marine water quality in the Florida Keys

    USGS Publications Warehouse

    Wetz, J.J.; Lipp, E.K.; Griffin, Dale W.; Lukasik, J.; Wait, D.; Sobsey, M.D.; Scott, T.M.; Rose, J.B.

    2004-01-01

    Concerns about the presence of enteric viruses in the surface waters of the Florida Keys prompted analyses of virus stability and persistence in these waters. In an in vitro study we evaluated the survival of poliovirus and stability of viral RNA in filtered natural seawater (FSW), unfiltered natural seawater (USW), artificial seawater (ASW) and DI water. This study compared cell culture infectivity with direct reverse transcription-polymerase chain reaction analysis. Attenuated poliovirus was seeded in the above water types and incubated in the dark at 22 and 30??C for 60 days. At 22??C, enhanced poliovirus survival and enhanced detection of viral RNA was observed in the seeded DI water control, artificial seawater and FSW samples. Detection of viruses in unfiltered seawater decreased rapidly at both temperatures by both methods of detection, suggesting that in the natural environment detection of enteroviral RNA may indicate a recent contamination event. In addition, in situ sampling in the Florida Keys during the late winter of 2000 revealed the presence of infectious enteroviruses at two sites and no sites exceeded recommended levels of microbial water quality indicators (enterococci or fecal coliform bacteria). ?? 2003 Elsevier Ltd. All rights reserved.

  12. DETECTION OF HUMAN ENTERIC VIRUSES IN STREAM WATER WITH RT-PCR AND CELL CULTURE

    EPA Science Inventory

    A multiplex RT-PCR method was used to measure virus occurrence at five stream water sites that span a range of hydroclimatic, water-quality, and land-use characteristics. The performance of the molecular method was evaluated in comparison to traditional cell culture and Escherich...

  13. A MULTI-LABORATORY EVALUATION OF METHODS FOR DETECTING ENTERIC VIRUSES IN SOILS

    EPA Science Inventory

    Two candidate methods for the recovery and detection of viruses in soil were subjected to round robin comparative testing by members of the American Society for Testing and Materials D19:24:04:04 Subcommittee Task Group. Selection of the methods, designated “Berg” and “Goyal,” wa...

  14. Presence of enteric viruses in source waters for drinking water production in The Netherlands.

    PubMed

    Lodder, W J; van den Berg, H H J L; Rutjes, S A; de Roda Husman, A M

    2010-09-01

    The quality of drinking water in The Netherlands has to comply with the Dutch Drinking Water Directive: less than one infection in 10,000 persons per year may occur due to consumption of unboiled drinking water. Since virus concentrations in drinking waters may be below the detection limit but entail a public health risk, the infection risk from drinking water consumption requires the assessment of the virus concentrations in source waters and of the removal efficiency of treatment processes. In this study, samples of source waters were taken during 4 years of regular sampling (1999 to 2002), and enteroviruses, reoviruses, somatic phages, and F-specific phages were detected in 75% (range, 0.0033 to 5.2 PFU/liter), 83% (0.0030 to 5.9 PFU/liter), 100% (1.1 to 114,156 PFU/liter), and 97% (0.12 to 14,403 PFU/liter), respectively, of 75 tested source water samples originating from 10 locations for drinking water production. By endpoint dilution reverse transcription-PCR (RT-PCR), 45% of the tested source water samples were positive for norovirus RNA (0.22 to 177 PCR-detectable units [PDU]/liter), and 48% were positive for rotavirus RNA (0.65 to 2,249 PDU/liter). Multiple viruses were regularly detected in the source water samples. A significant correlation between the concentrations of the two phages and those of the enteroviruses could be demonstrated. The virus concentrations varied greatly between 10 tested locations, and a seasonal effect was observed. Peak concentrations of pathogenic viruses occur in source waters used for drinking water production. If seasonal and short-term fluctuations coincide with less efficient or failing treatment, an unacceptable public health risk from exposure to this drinking water may occur.

  15. F+ RNA coliphage-based microbial source tracking in water resources of South Korea.

    PubMed

    Lee, Jung Eun; Lee, Heetae; Cho, You-Hee; Hur, Hor-Gil; Ko, GwangPyo

    2011-12-15

    We previously demonstrated that genotyping followed by proper statistical analyses of F plus (F+)-specific RNA coliphages can effectively represent fecal origins of either humans or animals. Here, we performed microbial source tracking (MST) using F+ RNA coliphages as a target MST microorganism for identifying fecal sources contaminating ground and surface water in metropolitan Seoul and Gyeonggi Province in South Korea. In total, 71 groundwater and 5 surface water samples were collected and screened for the presence of F+ RNA coliphages. More than 124 F+ coliphages were isolated from six groundwater and five surface water samples by the single agar layer method. F+ RNA coliphages were predominant in both waters (100% and 91%, respectively). Genotyping of 118 F+ RNA coliphages revealed that most (51/60) of the groundwater F+ RNA coliphages belonged to group I, whereas both groups I (25/58) and IV (31/58) were predominantly observed in surface water. Further comparison of phage isolates from human and animal (pig, cow, goose, and chicken) fecal sources using nucleic acid sequencing and principal coordinate analysis showed that groundwater samples formed clusters associated with cow feces, whereas surface waters formed clusters related to chicken and human feces. These results indicate the potential of the F+ RNA coliphage-based MST for identifying fecal contamination sources, which may be further exploited and validated in different geographical regions of the world.

  16. Specificity of coliphages in evaluating marker efficacy: a new insight for water quality indicators.

    PubMed

    Mookerjee, Subham; Batabyal, Prasenjit; Halder, Madhumanti; Palit, Anup

    2014-11-01

    Conventional procedures for qualitative assessment of coliphage are time consuming multiple step approach for achieving results. A modified and rapid technique has been introduced for determination of coliphage contamination among potable water sources during water borne outbreaks. During December 2013, 40 water samples from different potable water sources, were received for water quality analyses, from a jaundice affected Municipality of West Bengal, India. Altogether, 30% water samples were contaminated with coliform (1-20 cfu/ml) and 5% with E. coli (2-5 cfu/ml). Among post-outbreak samples, preponderance of coliform has decreased (1-4 cfu/ml) with total absence of E. coli. While standard technique has detected 55% outbreak samples with coliphage contamination, modified technique revealed that 80%, double than that of bacteriological identification rate, were contaminated with coliphages (4-20 pfu/10 ml). However, post-outbreak samples were detected with 1-5 pfu/10 ml coliphages among 20% samples. Coliphage detection rate through modified technique was nearly double (50%) than that of standard technique (27.5%). In few samples (with coliform load of 10-100 cfu/ml), while modified technique could detect coliphages among six samples (10-20 pfu/10 ml), standard protocol failed to detect coliphage in any of them. An easy, rapid and accurate modified technique has thereby been implemented for coliphage assessment from water samples. Coliform free water does not always signify pathogen free potable water and it is demonstrated that coliphage is a more reliable 'biomarker' to ascertain contamination level in potable water.

  17. Trypsin-treated Ma-104: a sensitive cell line for isolating enteric viruses from environmental samples.

    PubMed Central

    Agbalika, F; Hartemann, P; Foliguet, J M

    1984-01-01

    During a 1-year survey of enteroviruses in wastewater samples from the Lorraine area, three widely used continuous monkey kidney cell lines were tested: BGM, Vero, and trypsin-treated Ma-104. Decontaminated samples from secondary wastewater treatment plants (influent or effluent) were directly inoculated onto cells, and viruses were revealed after two passages with a liquid medium technique. Out of the total percentage of positive isolates with the three systems (32.7) 24.7% were found with Ma-104, 14.1% with BGM, and only 1.7% with Vero cells. Poliovirus was recovered more frequently with Ma-104 (12.3%) than with BGM (1.7%). Reovirus (3.5%) and echovirus (1.7%) were only found with Ma-104 cells; however, BGM cells allowed the isolation of a few group B coxsackieviruses (5.9%). It must be pointed out that 7.0% of samples with an unconfirmed cytopathic effect were found with BGM against 3.4% found with Ma-104, but they did not have significant differences. Because of its large spectrum of sensitivity, easy maintenance, and resistance to toxic effects, trypsin-treated Ma-104 may be recommended in conjunction with other cell lines for the detection of viruses from environmental samples, especially with the use of a liquid method. PMID:6324675

  18. Attenuated Salmonella Typhimurium delivery of a novel DNA vaccine induces immune responses and provides protection against duck enteritis virus.

    PubMed

    Liu, Xueyan; Liu, Qing; Xiao, Kangpeng; Li, Pei; Liu, Qiong; Zhao, Xinxin; Kong, Qingke

    2016-04-15

    DNA vaccines are widely used to prevent and treat infectious diseases, cancer and autoimmune diseases; however, their relatively low immunogenicity is an obstacle to their use. In this study, we constructed a novel and universal DNA vaccine vector (pSS898) that can be used to build DNA vaccines against duck enteritis virus (DEV) and other viruses that require DNA vaccines to provide protection. This vaccine vector has many advantages, including innate immunogenicity, efficient nuclear trafficking and resistance to attack from nucleases. UL24 and tgB from DEV were chosen as the antigens, and the heat labile enterotoxin B subunit (LTB) from Escherichia coli and the IL-2 gene (DuIL-2) from duck were used as adjuvants for the construction of DNA vaccine plasmids. Ducklings that were orally immunized with S739 (Salmonella Typhimurium Δasd-66 Δcrp-24 Δcya-25) and harboring these DEV DNA vaccines produced strong mucosal and systemic immune responses, and they resisted an otherwise lethal DEV challenge. More importantly, S739 (UL24-LTB) provided 90% protection after a priming-boost immunization. This study shows that our novel and universal DNA vaccine vector can be used efficiently in practical applications and may provide a promising method of orally inoculating ducks with a DEV DNA vaccine delivered by attenuated Salmonella Typhimurium for prevention of DVE.

  19. Use of dual priming oligonucleotide system-based multiplex RT-PCR combined with high performance liquid chromatography assay for simultaneous detection of five enteric viruses associated with acute enteritis.

    PubMed

    Fan, Wen-Lu; Wang, Zi-Wei; Qin, Yue; Sun, Chao; Liu, Zhong-Mei; Jiang, Yan-Ping; Qiao, Xin-Yuan; Tang, Li-Jie; Li, Yi-Jing; Xu, Yi-Gang

    2017-05-01

    In this study, a specific and sensitive method for simultaneous detection of human astrovirus, human rotavirus, norovirus, sapovirus and enteric adenovirus associated with acute enteritis was developed, based on the specific dual priming oligonucleotide (DPO) system and the sensitive high-performance liquid chromatography (HPLC) analysis. The DPO system-based multiplex reverse transcription-polymerase chain reaction (RT-PCR) combined with HPLC assay was more sensitive than agarose gel electrophoresis analysis and real-time SYBR Green PCR assay, and showed a specificity of 100% and sensitivity of 96%-100%. The high sensitivity and specificity of the assay indicates its great potential to be a useful tool for the accurate diagnosis of enteric virus infections.

  20. Duck enteritis virus glycoprotein D and B DNA vaccines induce immune responses and immunoprotection in Pekin ducks.

    PubMed

    Zhao, Yan; Cao, Yongsheng; Cui, Lihong; Ma, Bo; Mu, Xiaoyu; Li, Yanwei; Zhang, Zhihui; Li, Dan; Wei, Wei; Gao, Mingchun; Wang, Junwei

    2014-01-01

    DNA vaccine is a promising strategy for protection against virus infection. However, little is known on the efficacy of vaccination with two plasmids for expressing the glycoprotein D (gD) and glycoprotein B (gB) of duck enteritis virus (DEV) in inducing immune response and immunoprotection against virulent virus infection in Pekin ducks. In this study, two eukaryotic expressing plasmids of pcDNA3.1-gB and pcDNA3.1-gD were constructed. Following transfection, the gB and gD expressions in DF1 cells were detected. Groups of ducks were vaccinated with pcDNA3.1-gB and/or pcDNA3.1-gD, and boosted with the same vaccine on day 14 post primary vaccination. We found that intramuscular vaccinations with pcDNA3.1-gB and/or pcDNA3.1-gD, but not control plasmid, stimulated a high frequency of CD4+ and CD8+ T cells in Pekin ducks, particularly with both plasmids. Similarly, vaccination with these plasmids, particularly with both plasmids, promoted higher levels of neutralization antibodies against DEV in Pekin ducks. More importantly, vaccination with both plasmids significantly reduced the virulent DEV-induced mortality in Pekin ducks. Our data indicated that vaccination with plasmids for expressing both gB and gD induced potent cellular and humoral immunity against DEV in Pekin ducks. Therefore, this vaccination strategy may be used for the prevention of DEV infection in Pekin ducks.

  1. Leaching of viruses and other microorganisms naturally occurring in pig slurry to tile drains on a well-structured loamy field in Denmark

    NASA Astrophysics Data System (ADS)

    Krog, Jesper S.; Forslund, Anita; Larsen, Lars E.; Dalsgaard, Anders; Kjaer, Jeanne; Olsen, Preben; Schultz, Anna Charlotte

    2017-02-01

    The amount of animal manure used in modern agriculture is increasing due to the increase in global animal production. Pig slurry is known to contain zoonotic bacteria such as E. coli, Salmonella spp. and Campylobacter spp., and viruses such as hepatitis E virus and group A rotavirus. Coliform bacteria, present in manure, have previously been shown to leach into tile drains. This poses a potential threat to aquatic environments and may also influence the quality of drinking water. As knowledge is especially scarce about the fate of viruses when applied to fields in natural settings, this project sets out to investigate the leaching potential of six different microorganisms: E. coli and Enterococcus spp. (detected by colony assay), somatic coliphages (using plaque assays), and hepatitis E virus, porcine circovirus type 2, and group A rotavirus (by real-time polymerase chain reaction). All six microorganisms leached through the soil entering the tile drains situated at 1-m depth the first day following pig slurry application. The leaching pattern of group A rotavirus differed substantially from the pattern for somatic coliphages, which are otherwise used as indicators for virus contamination. Furthermore, group A rotavirus was detected in monitoring wells at 3.5-m depth up to 2 months after pig slurry application. The detection of viral genomic material in drainage water and shallow groundwater signifies a potential hazard to human health that needs to be investigated further, as water reservoirs used for recreational use and drinking water are potentially contaminated with zoonotic pathogens.

  2. Acute Diarrhea in West African Children: Diverse Enteric Viruses and a Novel Parvovirus Genus

    PubMed Central

    Phan, Tung G.; Vo, Nguyen P.; Bonkoungou, Isidore J. O.; Kapoor, Amit; Barro, Nicolas; O'Ryan, Miguel; Kapusinszky, Beatrix; Wang, Chunling

    2012-01-01

    Parvoviruses cause a variety of mild to severe symptoms or asymptomatic infections in humans and animals. During a viral metagenomic analysis of feces from children with acute diarrhea in Burkina Faso, we identified in decreasing prevalence nucleic acids from anelloviruses, dependoviruses, sapoviruses, enteroviruses, bocaviruses, noroviruses, adenoviruses, parechoviruses, rotaviruses, cosavirus, astroviruses, and hepatitis B virus. Sequences from a highly divergent parvovirus, provisionally called bufavirus, were also detected whose NS1 and VP1 proteins showed <39% and <31% identities to those of previously known parvoviruses. Four percent of the fecal samples were PCR positive for this new parvovirus, including a related bufavirus species showing only 72% identity in VP1. The high degree of genetic divergence of these related genomes from those of other parvoviruses indicates the presence of a proposed new Parvoviridae genus containing at least two species. Studies of the tropism and pathogenicity of these novel parvoviruses will be facilitated by the availability of their genome sequences. PMID:22855485

  3. Acute diarrhea in West African children: diverse enteric viruses and a novel parvovirus genus.

    PubMed

    Phan, Tung G; Vo, Nguyen P; Bonkoungou, Isidore J O; Kapoor, Amit; Barro, Nicolas; O'Ryan, Miguel; Kapusinszky, Beatrix; Wang, Chunling; Delwart, Eric

    2012-10-01

    Parvoviruses cause a variety of mild to severe symptoms or asymptomatic infections in humans and animals. During a viral metagenomic analysis of feces from children with acute diarrhea in Burkina Faso, we identified in decreasing prevalence nucleic acids from anelloviruses, dependoviruses, sapoviruses, enteroviruses, bocaviruses, noroviruses, adenoviruses, parechoviruses, rotaviruses, cosavirus, astroviruses, and hepatitis B virus. Sequences from a highly divergent parvovirus, provisionally called bufavirus, were also detected whose NS1 and VP1 proteins showed <39% and <31% identities to those of previously known parvoviruses. Four percent of the fecal samples were PCR positive for this new parvovirus, including a related bufavirus species showing only 72% identity in VP1. The high degree of genetic divergence of these related genomes from those of other parvoviruses indicates the presence of a proposed new Parvoviridae genus containing at least two species. Studies of the tropism and pathogenicity of these novel parvoviruses will be facilitated by the availability of their genome sequences.

  4. Rift Valley fever virus strain MP-12 enters mammalian host cells via caveola-mediated endocytosis.

    PubMed

    Harmon, Brooke; Schudel, Benjamin R; Maar, Dianna; Kozina, Carol; Ikegami, Tetsuro; Tseng, Chien-Te Kent; Negrete, Oscar A

    2012-12-01

    Rift Valley fever virus (RVFV) is a zoonotic pathogen capable of causing serious morbidity and mortality in both humans and livestock. The lack of efficient countermeasure strategies, the potential for dispersion into new regions, and the pathogenesis in humans and livestock make RVFV a serious public health concern. The receptors, cellular factors, and entry pathways used by RVFV and other members of the family Bunyaviridae remain largely uncharacterized. Here we provide evidence that RVFV strain MP-12 uses dynamin-dependent caveola-mediated endocytosis for cell entry. Caveolae are lipid raft domains composed of caveolin (the main structural component), cholesterol, and sphingolipids. Caveola-mediated endocytosis is responsible for the uptake of a wide variety of host ligands, as well as bacteria, bacterial toxins, and a number of viruses. To determine the cellular entry mechanism of RVFV, we used small-molecule inhibitors, RNA interference (RNAi), and dominant negative (DN) protein expression to inhibit the major mammalian cell endocytic pathways. Inhibitors and RNAi specific for macropinocytosis and clathrin-mediated endocytosis had no effect on RVFV infection. In contrast, inhibitors of caveola-mediated endocytosis, and RNAi targeted to caveolin-1 and dynamin, drastically reduced RVFV infection in multiple cell lines. Expression of DN caveolin-1 also reduced RVFV infection significantly, while expression of DN EPS15, a protein required for the assembly of clathrin-coated pits, and DN PAK-1, an obligate mediator of macropinocytosis, had no significant impact on RVFV infection. These results together suggest that the primary mechanism of RVFV MP-12 uptake is dynamin-dependent, caveolin-1-mediated endocytosis.

  5. Human and animal enteric virus in groundwater from deep wells, and recreational and network water.

    PubMed

    Fongaro, Gislaine; Padilha, J; Schissi, C D; Nascimento, M A; Bampi, G B; Viancelli, A; Barardi, C R M

    2015-12-01

    This study was designed to assess the presence of human adenovirus (HAdV), rotavirus-A (RVA), hepatitis A virus (HAV), and porcine circovirus-2 (PCV2) in groundwater from deep wells, and recreational and network waters. The water samples were collected and concentrated and the virus genomes were assessed and quantified by quantitative PCR (qPCR). Infectious HAdV was evaluated in groundwater and network water samples by integrated cell culture using transcribed messenger RNA (mRNA) (ICC-RT-qPCR). In recreational water samples, HAdV was detected in 100 % (6/6), HAV in 66.6 % (4/6), and RVA in 66.6 % (4/6). In network water, HAdV was detected in 100 % (6/6) of the samples (these 83 % contained infectious HAdV), although HAV and RVA were not detected and PCV2 was not evaluated. In groundwater from deep wells, during rainy period, HAdV and RVA were detected in 80 % (4/5) of the samples, and HAV and PCV2 were not detected; however, during dry period, HAdV and RVA were detected in 60 % (3/5), HAV in only one sample, and PCV2 in 60 % (4/5). In groundwater, all samples contained infectious HAdV. PCV2 presence in groundwater is indicative of contamination caused by swine manure in Concórdia, Santa Catarina, Brazil. The disinfection of human and animal wastes is urgent, since they can contaminate surface and groundwater, being a potential threat for public and animal health.

  6. Occurrence and distribution of enteric viruses in shallow ground water and factors affecting well vulnerability to microbiological contamination in Worcester and Wicomico counties, Maryland

    USGS Publications Warehouse

    Banks, William S.L.; Klohe, Cheryl A.; Battigelli, David A.

    2001-01-01

    The U.S. Geological Survey, in cooperation with the Maryland Department of the Environment and the Wisconsin State Laboratory of Hygiene, conducted a study to characterize the occurrence and distribution of viral contamination in small (withdrawing less than 10,000 gallons per day) public water-supply wells screened in the water-table aquifer in the Coastal Plain in Worcester and Wicomico Counties, Maryland.Two hundred seventy-eight well sites were evaluated with regard to simulated ground-water flow paths, land use, natural soils groups, and well characteristics, such as well depth and well age. Flow and transport simulations of the water-table aquifer indicated that wells screened less than about 50 feet below land surface (shallow wells) were most vulnerable to surface contamination, which in some cases could originate from as far as 2,000 feet upgradient of the well. Animal-feeding and agricultural-storage operations were considered among the most likely sources for viral contamination; therefore, sites close to these activities were considered most vulnerable. Soil groups were evaluated with regard to depth to water and moisture-holding capacity. Wells with shallow depths to water or in very sandy soils were considered more vulnerable to contamination than deep wells (greater than 50 feet) and those completed in finer-grained soils. Older wells and wells where coliform bacteria had been detected in the past were classified as highly vulnerable. On the basis of this evaluation, 27 sites considered to be susceptible were sampled.Samples were collected by pumping up to 400 gallons of untreated well water through an electropositive filter. Water concentrates were subjected to cell-culture assay for the detection of culturable viruses and reverse-transcription polymerase chain reaction/gene probe assays to detect nonculturable viruses; grab samples were analyzed for somatic and male-specific coliphages, Bacteroides fragilis, Clostridium perfringens, enterococci

  7. A case study of enteric virus removal and insights into the associated risk of water reuse for two wastewater treatment pond systems in Bolivia.

    PubMed

    Symonds, E M; Verbyla, M E; Lukasik, J O; Kafle, R C; Breitbart, M; Mihelcic, J R

    2014-11-15

    Wastewater treatment ponds (WTP) are one of the most widespread treatment technologies in the world; however, the mechanisms and extent of enteric virus removal in these systems are poorly understood. Two WTP systems in Bolivia, with similar overall hydraulic retention times but different first stages of treatment, were analyzed for enteric virus removal. One system consisted of a facultative pond followed by two maturation ponds (three-pond system) and the other consisted of an upflow anaerobic sludge blanket (UASB) reactor followed by two maturation (polishing) ponds (UASB-pond system). Quantitative polymerase chain reaction with reverse transcription (RT-qPCR) was used to measure concentrations of norovirus, rotavirus, and pepper mild mottle virus, while cell culture methods were used to measure concentrations of culturable enteroviruses (EV). Limited virus removal was observed with RT-qPCR in either system; however, the three-pond system removed culturable EV with greater efficiency than the UASB-pond system. The majority of viruses were not associated with particles and only a small proportion was associated with particles larger than 180 μm; thus, it is unlikely that sedimentation is a major mechanism of virus removal. High concentrations of viruses were associated with particles between 0.45 and 180 μm in the UASB reactor effluent, but not in the facultative pond effluent. The association of viruses with this size class of particles may explain why only minimal virus removal was observed in the UASB-pond system. Quantitative microbial risk assessment of the treated effluent for reuse for restricted irrigation indicated that the three-pond system effluent requires an additional 1- to 2-log10 reduction of viruses to achieve the WHO health target of <10(-4) disability-adjusted life years (DALYs) lost per person per year; however, the UASB-pond system effluent may require an additional 2.5- to 4.5-log10 reduction of viruses.

  8. The failure of an inactivated mink enteritis virus vaccine in four preparations to provide protection to dogs against challenge with canine parvovirus-2.

    PubMed Central

    Carman, S; Povey, C

    1982-01-01

    Four experimental vaccine preparations comprising a strain of mink enteritis virus inactivated by either formalin or beta-propiolactone, and either adjuvanted or nonadjuvanted, failed to stimulate a consistent serum antibody response in 20 vaccinated dogs and failed to protect all but one of these dogs against oral challenge with canine parvovirus-2. PMID:6280820

  9. Endocytic Pathways Used by Andes Virus to Enter Primary Human Lung Endothelial Cells

    PubMed Central

    Flint, Mike; Lin, Jin-Mann S.; Spiropoulou, Christina F.

    2016-01-01

    Andes virus (ANDV) is the major cause of hantavirus pulmonary syndrome (HPS) in South America. Despite a high fatality rate (up to 40%), no vaccines or antiviral therapies are approved to treat ANDV infection. To understand the role of endocytic pathways in ANDV infection, we used 3 complementary approaches to identify cellular factors required for ANDV entry into human lung microvascular endothelial cells. We screened an siRNA library targeting 140 genes involved in membrane trafficking, and identified 55 genes required for ANDV infection. These genes control the major endocytic pathways, endosomal transport, cell signaling, and cytoskeleton rearrangement. We then used infectious ANDV and retroviral pseudovirions to further characterize the possible involvement of 9 of these genes in the early steps of ANDV entry. In addition, we used markers of cellular endocytosis along with chemical inhibitors of known endocytic pathways to show that ANDV uses multiple routes of entry to infect target cells. These entry mechanisms are mainly clathrin-, dynamin-, and cholesterol-dependent, but can also occur via a clathrin-independent manner. PMID:27780263

  10. Quantification of Enteric Viruses, Pathogen Indicators, and Salmonella Bacteria in Class B Anaerobically Digested Biosolids by Culture and Molecular Methods ▿

    PubMed Central

    Wong, Kelvin; Onan, Brandon M.; Xagoraraki, Irene

    2010-01-01

    The most common class B biosolids in the United States are generated by mesophilic anaerobic digestion (MAD), and MAD biosolids have been used for land application. However, the pathogen levels in MAD biosolids are still unclear, especially with respect to enteric viruses. In this study, we determined the occurrence and the quantitative levels of enteric viruses and indicators in 12 MAD biosolid samples and of Salmonella enterica in 6 MAD biosolid samples. Three dewatered biosolid samples were also included in this study for purposes of comparison. Human adenoviruses (HAdV) had the highest gene levels and were detected more frequently than other enteric viruses. The gene levels of noroviruses (NV) reported were comparable to those of enteroviruses (EV) and human polyomaviruses (HPyV). The occurrence percentages of HAdV, HAdV species F, EV, NV GI, NV GII, and HPyV in MAD samples were 83, 83, 42, 50, 75, and 58%, respectively. No hepatitis A virus was detected. Infectious HAdV was detected more frequently than infectious EV, and all infectious HAdV were detected when samples were propagated in A549 cells. Based on most-probable-number (MPN) analysis, A549 cells were more susceptible to biosolid-associated viruses than BGM cells. All indicator levels in MAD biosolids were approximately 104 MPN or PFU per gram (dry), and the dewatered biosolids had significantly higher indicator levels than the MAD biosolids. Only two MAD samples tested positive for Salmonella enterica, where the concentration was below 1.0 MPN/4 g. This study provides a broad comparison of the prevalence of different enteric viruses in MAD biosolids and reports the first detection of noroviruses in class B biosolids. The observed high quantitative and infectivity levels of adenoviruses in MAD biosolids indicate that adenovirus is a good indicator for the evaluation of sludge treatment efficiency. PMID:20693452

  11. Efficacy study and field application of an inactivated new type gosling viral enteritis virus vaccine for domestic geese.

    PubMed

    Chen, S; Ma, G P; Wang, M S; Cheng, A C; Zhu, D K; Luo, Q H; Jia, R Y; Liu, F; Chen, X Y; Han, X F; Bo, Y; Zhou, D C

    2011-04-01

    New type gosling viral enteritis virus (NGVEV) caused a serious disease in naive juvenile goslings. In the described studies the performance of 2 vaccines was analyzed: a vaccine containing adjuvanted inactivated NGVEV and a vaccine containing adjuvanted inactivated NGVEV and recombinant goose IL-2. Breeder geese were subcutaneously vaccinated at the beginning of the egg production period with the vaccines. Breeder geese sham vaccinated with PBS served as control. The cellular and humoral immune responses of the vaccinated breeder geese, as well as the presence of maternally derived antibody to NGVEV, were investigated by ELISA, virus neutralization test, and lymphocyte proliferation assay, respectively. A significantly higher immunogenicity (P < 0.05) was induced by the inactivated NGVEV-recombinant goose IL-2 adjuvant vaccine compared with the inactivated NGVEV vaccine. The offspring of the vaccinated birds were challenged with virulent NGVEV (100 50% lethal dose) and the protective efficacy of the vaccines was determined. Furthermore, in a field trial the efficacy of the inactivated NGVEV vaccine was recorded from years 2003 to 2007. No clinical signs or abnormal health status were observed in the vaccinated breeder geese and the progeny. After a single application, >80% protection was shown in the progeny of geese vaccinated against NGVEV challenge for approximately 5 mo. The extensive field trials further demonstrated that vaccination of breeder geese with the inactivated NGVEV vaccine could be a safe and efficacious means to control NGVE disease. Moreover, the level of maternally derived NGVEV antibody titer in the egg yolk reflected the level of NGVEV antibodies in the breeder geese, suggesting that the egg yolk could be used to monitor the vaccination efficacy in commercial goose breeder flocks.

  12. The transcription analysis of duck enteritis virus UL49.5 gene using real-time quantitative reverse transcription PCR.

    PubMed

    Lin, Meng; Jia, Renyong; Wang, Mingshu; Gao, Xinghong; Zhu, Dekang; Chen, Shun; Yin, Zhongqiong; Wang, Yin; Chen, Xiaoyue; Cheng, Anchun

    2013-10-01

    Duck enteritis virus (DEV) UL49.5 encoding glycoprotein N was a conserved gene. The transcription dynamic process of UL49.5 homologous genes in herpesviruses was reported. However, the transcription dynamic process of DEV UL49.5 gene has not yet been established. In this study, a real-time quantitative reverse transcription PCR (real-time qRT-PCR) assay was established to test the transcription dynamic process of DEV UL49.5 gene, and the recombinant plasmid pUCm-T/UL49.5 was constructed as the standard DNA. The samples prepared from DEV-infected (at different time points) and uninfected cell were detected and calculated. The results demonstrated that the real-time qRT-PCR assay was successfully established. The transcription product of DEV UL49.5 gene was first detected at 0.5 h post infection (p.i.), increased at 8 h p.i. and reached a peak at 60 h p.i. Our results illustrated that DEV UL49.5 gene could be regarded as a late gene. The transcription dynamic process of DEV UL49.5 gene may provide a significant clue for further studies of DEV UL49.5 gene.

  13. Coliphages and bacteria in ground water from Tehran, Iran

    SciTech Connect

    Shariatpanahi, M.; Anderson, A.C.

    1987-07-01

    The purpose of this study was to examine the microbial quality of Tehran's ground water and selected springs, using coliphages and selected bacteria as indicator organisms. The water table in Tehran varies from approximately 160 meters in the north to approximately 5 meters in the south. Individual wells and subterranean man-made aqueducts (qanate) tap the ground water. Since Tehran lacks municipal sewage facilities, waste disposal is by means of seepage pits, privies and leaching cesspools. There is potential for waste from these sites to leach into the ground water, particularly in the south where the water table is near the surface and the clay content of the soil holds moisture during periods of heavy rainfall.

  14. Somatic coliphages as surrogates for enteroviruses in sludge hygienization treatments.

    PubMed

    Martín-Díaz, Julia; Casas-Mangas, Raquel; García-Aljaro, Cristina; Blanch, Anicet R; Lucena, Francisco

    2016-01-01

    Conventional bacterial indicators present serious drawbacks giving information about viral pathogens persistence during sludge hygienization treatments. This calls for the search of alternative viral indicators. Somatic coliphages' (SOMCPH) ability for acting as surrogates for enteroviruses was assessed in 47 sludge samples subjected to novel treatment processes. SOMCPH, infectious enteroviruses and genome copies of enteroviruses were monitored. Only one of these groups, the bacteriophages, was present in the sludge at concentrations that allowed the evaluation of treatment's performance. An indicator/pathogen relationship of 4 log10 (PFU/g dw) was found between SOMCPH and infective enteroviruses and their detection accuracy was assessed. The obtained results and the existence of rapid and standardized methods encourage the inclusion of SOMCPH quantification in future sludge directives. In addition, an existing real-time quantitative polymerase chain reaction (RT-qPCR) for enteroviruses was adapted and applied.

  15. Development of a feasible method to extract somatic coliphages from sludge, soil and treated biowaste.

    PubMed

    Guzmán, Carolina; Jofre, Juan; Blanch, Anicet R; Lucena, Francisco

    2007-09-01

    Extraction of viruses and bacteriophages from sludge, soil and treated biowaste requires homogenization, elution, clarification and detoxification-decontamination steps. Seeding these matrixes with bacteriophages does not reproduce what happens in nature. Therefore, naturally occurring matrixes, raw sludge, digested and dewatered sludge and compost, containing high numbers of somatic coliphages, and soils contaminated with wastewater or raw sludge were used in the extraction assays. Based on eluting the bacteriophages with beef extract, a feasible method in which the different steps had been optimized has been established. The method is feasible, repeatable, robust and applicable in routine laboratories. Digested and dewatered sludge has been probed to be useful as a reference material for validation studies and for "in lab" quality control. The established method includes homogenization by magnetic stirring, elution (which is performed at the same time that homogenization) with 10% beef extract at neutral pH, clarification by centrifuging at 4000 x g and decontamination by filtration through low protein binding 0.22 microm diameter pore size membrane filters.

  16. Inactivation of MS2 coliphage by UV and hydrogen peroxide: comparison by cultural and molecular methodologies.

    PubMed

    Sherchan, Samendra P; Snyder, Shane A; Gerba, Charles P; Pepper, Ian L

    2014-01-01

    The use of advanced oxidation processes (AOP) are expected to increase for removal of emerging contaminants and pathogens from drinking water. In this study, the performance of a small community ultraviolet light reactor in combination with hydrogen peroxide (H2O2) for MS2 coliphage inactivation with two different flow rate conditions of 1 gal/min (gpm) and 2 gpm was evaluated. Following UV radiation, MS2 showed a reduction of 5.3-5.8 log10 when quantified with cultural plaque counts, whereas corresponding quantitative polymerase chain reaction (qPCR) data showed only a 1.7-2.8 log10 reduction in viral RNA copy number. When H2O2 was added at either 2.5 or 5 ppm with UV at both flow rate conditions, enhanced MS2 inactivation occurred with a more than 7 log10 reduction observed via plaque counts, indicating that all added MS2 had been inactivated, since no plaques were formed after incubation at 37 °C for 24 h. In contrast, qPCR only showed a corresponding 3-4 log10 reduction in viral RNA copy number. This research also sheds light on the inactivation of MS2 with ultraviolet light and in the presence of hydroxyl radicals and provides a practical use of qPCR to detect MS2 concentration following advanced oxidation relative to traditional plaque methodology; however qPCR detection overestimates the true number of infective virus.

  17. Genotyping male-specific RNA coliphages by hybridization with oligonucleotide probes.

    PubMed Central

    Hsu, F C; Shieh, Y S; van Duin, J; Beekwilder, M J; Sobsey, M D

    1995-01-01

    F-specific (F+) RNA coliphages are prevalent in sewage and other fecal wastes of humans and animals. There are four antigenically distinct serogroups of F+ RNA coliphages, and those predominating in humans (groups II and III) differ from those predominating in animals (groups I and IV). Hence, it may be possible to distinguish between human and animal wastes by serotyping F+ RNA coliphage isolates. Because serotyping is laborious and requires scarce antiserum reagents, we investigated genotyping using synthetic oligonucleotide probes as an alternative approach to distinguishing the four groups of F+ RNA coliphages. Oligoprobes I, II, III, IV, A, and B were selected to detect group I, II, III, IV, I plus II, and III plus IV phages, respectively. Methods for phage transfer from zones of lysis on a host cell lawn to candidate membrane filters and fixation of genomic nucleic acid on the membranes were optimized. The oligoprobes, which were end labeled with digoxigenin, were applied in DNA-RNA hybridization, and hybrids were observed by colorimetric, immunoenzymatic detection. Of 203 isolates of F+ RNA coliphages from environmental samples of water, wastes, and shellfish, 99.5 and 96.6% could be classified into each group by serotyping and genotyping, respectively. Probes A and B correctly identified 100% of the isolates. On the basis of these results, this method for genotyping F+ RNA coliphages appears to be practical and reliable for typing isolates in field samples. PMID:8526509

  18. Virulence Factors and Stability of Coliphages Specific to Escherichia coli O157:H7 and to Various E. coli Infection.

    PubMed

    Kim, Eun-Jin; Chang, Hyun-Joo; Kwak, Soojin; Park, Jong-Hyun

    2016-12-28

    Characteristics of E. coli O157:H7-specific infection bacteriophages (O157 coliphages) and broad-host-range bacteriophages for other E. coli serotypes (broad-host coliphages) were compared. The burst sizes of the two groups ranged from 40 to 176 PFU/infected cell. Distributions of the virulence factors stx1, stx2, ehxA, and saa between the two groups were not differentiated. Broad-host-range coliphages showed lower stability at 70°C, in relation to O157 coliphages. However, O157 coliphages showed high acid and ethanol tolerance by reduction of only 22% and 11% phages, respectively, under pH 3 and 70% ethanol for 1 h exposure. Therefore, these results revealed that the O157 coliphages might be more stable under harsh environments, which might explain their effective infection of the acid-tolerant E. coli O157:H7.

  19. Investigation of enteric adenovirus and poliovirus removal by coagulation processes and suitability of bacteriophages MS2 and φX174 as surrogates for those viruses.

    PubMed

    Shirasaki, N; Matsushita, T; Matsui, Y; Marubayashi, T; Murai, K

    2016-09-01

    We evaluated the removal of enteric adenovirus (AdV) type 40 and poliovirus (PV) type 1 by coagulation, using water samples from 13 water sources for drinking water treatment plants in Japan. The behaviors of two widely accepted enteric virus surrogates, bacteriophages MS2 and φX174, were compared with the behaviors of AdV and PV. Coagulation with polyaluminum chloride (PACl, basicity 1.5) removed AdV and PV from virus-spiked source waters: the infectious AdV and PV removal ratios evaluated by means of a plaque-forming-unit method were 0.1-1.4-log10 and 0.5-2.4-log10, respectively. A nonsulfated high-basicity PACl (basicity 2.1) removed infectious AdV and PV more efficiently than did other commercially available PACls (basicity 1.5-2.1), alum, and ferric chloride. The MS2 removal ratios tended to be larger than those of AdV and PV, partly because of differences in the hydrophobicities of the virus particles and the sensitivity of the virus to the virucidal activity of PACl; the differences in removal ratios were not due to differences in the surface charges of the virus particles. MS2, which was more hydrophobic than the other viruses, was inactivated during coagulation with PACl. Therefore, MS2 does not appear to be an appropriate surrogate for AdV and PV during coagulation. In contrast, because φX174, like AdV and PV, was not inactivated during coagulation, and because the hydrophobicity of φX174 was similar to or somewhat lower than the hydrophobicities of AdV and PV, the φX174 removal ratios tended to be similar to or somewhat smaller than those of the enteric viruses. Therefore, φX174 is a potential conservative surrogate for AdV and PV during coagulation. In summary, the surface hydrophobicity of virus particles and the sensitivity of the virus to the virucidal activity of the coagulant are probably important determinants of the efficiency of virus removal during coagulation.

  20. Virus and Bacteria Removal from Wastewater by Rapid Infiltration Through Soil

    PubMed Central

    Schaub, Stephen A.; Sorber, Charles A.

    1977-01-01

    A rapid infiltration land wastewater application site, composed of unconsolidated silty sand and gravel, which has been in continuous operation for over 30 years was examined for the accumulation and/or migration of a tracer virus (coliphage f2), indigenous enteroviruses, and enteric indicator bacteria in the soils and underlying groundwater. Tracer f2 penetrated into groundwater together with the front of percolating primary effluent and was not observed to concentrate on the upper soil layers. The tracer virus concentration in a 60-foot (about 18.3-m)-deep observation well directly beneath the wastewater application area began to increase within 48 h after application to the soil. The tracer level in this well stabilized after 72 h at a level of approximately 47% of the average applied concentration. Indigenous enteroviruses and tracer f2 were sporadically detected in the groundwater at horizontal distances of 600 feet (about 183 m) from the application zone. Laboratory soil adsorption studies confirmed the poor virus adsorption observed at the site. This was especially true on surface soils when contained in wastewater. Enteric indicator bacteria were readily concentrated on the soil surface by filtration on the soil surface mat. However, during tracer f2 virus tests, comparison studies with fecal Streptococcus revealed that bacteria capable of penetrating the surface were able to migrate into the groundwater. They were detected at the same locations as tracer and enteric viruses. PMID:16345215

  1. Bioaerosol Dispersion in Relation with Wastewater Reuse for Crop Irrigation. (Experiments to understand emission processes with enteric virus and risks modeling).

    NASA Astrophysics Data System (ADS)

    Courault, D.; Girardin, G.; Capowiez, L.; Albert, I.; Krawczyk, C.; Ball, C.; Salemkour, A.; Bon, F.; Perelle, S.; Fraisse, A.; Renault, P.; Amato, P.

    2014-12-01

    Bio-aerosols consist of microorganisms or biological particles that become airborne depending on various environmental factors. Recycling of wastewater (WW) for irrigation can cope with the issues of water availability, and it can also threaten Human health if the pathogens present in WW are aerosolized during sprinkling irrigation or wind events. Among the variety of micro-organisms found in WW, enteric viruses can reach significant amounts, because most of the WW treatments are not completely efficient. These viruses are particularly resistant in the environment and responsibles of numerous digestive diseases (gastroenteritis, hepatitis…). Few quantities are enough to make people sick (102 pfu). Several knowledge gaps exist to better estimate the risks for Human exposure, and on the virus transfer from irrigation up to the respiratory track. A research program funded by the French government (INSU), gathering multi disciplinary teams aims at better understanding virus fate in air and health risks from WW reuse. Experiments were conducted under controlled conditions in order to prioritize the main factors impacting virus aerosolization. Irrigation with water loaded with safe surrogates of Hepatitis A virus (Murine Mengo Virus) was applied on small plots covered by channels in which the wind speed varied. Various situations have been investigated (wet/dry surfaces, strong/mild winds, clean/waste water). Air samples were collected above plots using impingers and filters after irrigation for several days. Viruses were quantified by RT-qPCR. The results showed that impingers were more efficient in airborne virus recovering than filters. Among environmental factors, Wind speed was the main factor explaining virus concentration in the air after irrigation. A Quantitative Microbial Risk Assessment approach has been chosen to assess the health effects on the population. The main modeling steps will be presented, including a simplified dispersion model coupled with a

  2. Enhanced enteric virus detection in sporadic gastroenteritis using a multi-target real-time PCR panel: a one-year study.

    PubMed

    Pang, Xiaoli L; Preiksaitis, Jutta K; Lee, Bonita E

    2014-09-01

    Viral gastroenteritis causes significant mortality and morbidity worldwide. Identifying the etiology of viral gastroenteritis is a challenge as most enteric viruses (EVs) are non-culturable. This study is to develop an EV testing panel using real-time PCR (EVPrtPCR) to simultaneously detect rotavirus, norovirus, sapovirus, astrovirus, and enteric adenovirus in stool samples. EVPrtPCR using universal amplification conditions was run in a single instrument run. EVPrtPCR was used to test 2,486 sporadic gastroenteritis samples submitted for EV testing using electron microscopy (EM) between July 2008 and July 2009. Retesting spiked negative stool samples and Salmon DNA as internal control were used to evaluate inhibition. EVPrtPCR detected viruses in significantly more samples: 748 (34%) as compared to 94 (3.8%) by EM. EM did not detect any norovirus, sapovirus, and mixed infection, and detected only 39% of rotavirus and 38.2% of enteric adenovirus positive samples. Four samples that tested positive for rotavirus and two for adenovirus and for small-round-structured viruses by EM were negative by EVPrtPCR. Norovirus was the most common virus detected (17.6%) with 92.4% as genogroup II, followed by rotavirus (6.8%), sapovirus (4.2%), astrovirus (2.0%), and enteric adenovirus (1.4%) with 9% samples positive for mixed infection. Overall, EV identification followed a U-shaped age distribution; positive samples were more common in children ≤5 years old and adults >60 years old. Norovirus, sapovirus and astrovirus showed winter predominance and rotavirus peaked in the spring. No inhibition was observed. Molecular technology significantly enhanced the identification of EV causing sporadic gastroenteritis in Alberta.

  3. Response of mink, skunk, red fox and raccoon to inoculation with mink virus enteritis, feline panleukopenia and canine parvovirus and prevalence of antibody to parvovirus in wild carnivores in Ontario.

    PubMed Central

    Barker, I K; Povey, R C; Voigt, D R

    1983-01-01

    Mink virus enteritis, feline panleukopenia and canine parvovirus-2 were inoculated separately into groups of raccoon, mink, red fox and striped skunk. Raccoons were highly susceptible to mink virus enteritis and feline panleukopenia, with animals developing clinical illness, and several dying within six to ten days of inoculation with lesions typical of parvovirus infection. Both viruses were shed in high titre in the feces of infected raccoons, and high antibody titres were stimulated. Raccoons inoculated with canine parvovirus-2 showed no signs; shedding of virus was sporadic though moderate titres of antibody developed. Mink inoculated with mink virus enteritis and feline panleukopenia developed signs and lesions of early parvovirus infection. No signs or significant lesions followed canine parvovirus-2 inoculation. Shedding of virus was heavy (mink virus enteritis) or sporadic (feline panleukopenia and canine parvovirus-2), though good serological responses were elicited to all three viruses. Red fox showed no signs of infection, shed all three viruses only sporadically, and the serological response was strong only to feline panleukopenia. Skunks developed low antibody titres, but no signs, and did not shed virus. Antibody to parvovirus was found in 79.2% of 144 wild red foxes; 22.3% of 112 wild raccoons; 1.3% of 157 wild skunks and 6/7 coyotes in southern Ontario. The likely significance of these viruses to wild and captive individuals and populations of these carnivores is discussed. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. PMID:6309349

  4. Occurrence of water-borne enteric viruses in two settlements based in Eastern Chad: analysis of hepatitis E virus, hepatitis A virus and human adenovirus in water sources.

    PubMed

    Guerrero-Latorre, Laura; Carratala, Anna; Rodriguez-Manzano, Jesus; Calgua, Byron; Hundesa, Ayalkibet; Girones, Rosina

    2011-09-01

    Hepatitis E virus (HEV) is a common cause of water-borne acute hepatitis in areas with poor sanitation. In 2004 an outbreak of HEV infection affected around 2,000 people in Eastern Chad (Dar Sila). This paper describes the decrease in the incidence of acute jaundice syndrome (AJS) from 2004 until 2009 when a mean incidence of 0.48 cases/1,000 people/year was recorded in the region. Outbreaks of AJS were identified in some of the camps in 2007 and 2008. Moreover, water samples from drinking water sources were screened for human adenoviruses considered as viral indicators and for hepatitis A virus and HEV. Screening of faecal samples from donkeys for HEV gave negative results. Some of the samples were also analysed for faecal coliforms showing values before disinfection treatment between 3 and >50 colony forming units per 100 mL. All water samples tested were negative for HEV and HAV; however, the presence of low levels of human adenoviruses in 4 out of 16 samples analysed indicates possible human faecal contamination of groundwater. Consequently, breakdowns in the treatment of drinking water and/or increased excretion of hepatitis viruses, which could be related to the arrival of a new population, could spread future outbreaks through drinking water.

  5. Organic Wastewater Compounds, Pharmaceuticals, andColiphage in Ground Water Receiving Discharge from OnsiteWastewater Treatment Systems near La Pine, Oregon:Occurrence and Implications for Transport

    USGS Publications Warehouse

    Hinkle, Stephen J.; Weick, Rodney J.; Johnson, Jill M.; Cahill, Jeffery D.; Smith, Steven G.; Rich, Barbara J.

    2005-01-01

    The occurrence of organic wastewater compounds (components of 'personal care products' and other common household chemicals), pharmaceuticals (human prescription and nonprescription medical drugs), and coliphage (viruses that infect coliform bacteria, and found in high concentrations in municipal wastewater) in onsite wastewater (septic tank effluent) and in a shallow, unconfined, sandy aquifer that serves as the primary source of drinking water for most residents near La Pine, Oregon, was documented. Samples from two types of observation networks provided basic occurrence data for onsite wastewater and downgradient ground water. One observation network was a group of 28 traditional and innovative (advanced treatment) onsite wastewater treatment systems and associated downgradient drainfield monitoring wells, referred to as the 'innovative systems network'. The drainfield monitoring wells were located adjacent to or under onsite wastewater treatment system drainfield lines. Another observation network, termed the 'transect network', consisted of 31 wells distributed among three transects of temporary, stainless-steel-screened, direct-push monitoring wells installed along three plumes of onsite wastewater. The transect network, by virtue of its design, also provided a basis for increased understanding of the transport of analytes in natural systems. Coliphage were frequently detected in onsite wastewater. Coliphage concentrations in onsite wastewater were highly variable, ranging from less than 1 to 3,000,000 plaque forming units per 100 milliliters. Coliphage were occasionally detected (eight occurrences) at low concentrations in samples from wells located downgradient from onsite wastewater treatment system drainfield lines. However, coliphage concentrations were below method detection limits in replicate or repeat samples collected from the eight sites. The consistent absence of coliphage detections in the replicate or repeat samples is interpreted to indicate

  6. Detection of somatic phages, infectious enteroviruses and enterovirus genomes as indicators of human enteric viral pollution in surface water.

    PubMed

    Hot, D; Legeay, O; Jacques, J; Gantzer, C; Caudrelier, Y; Guyard, K; Lange, M; Andréoletti, L

    2003-11-01

    In the present study, we aimed to determine whether the concentrations of somatic coliphages, infectious enteroviruses or the detection of enterovirus genomes were associated with the detection of human pathogenic viruses in surface water. Four French rivers were sampled monthly or semimonthly for the quantitative detection of somatic coliphages, infectious enteroviruses and the qualitative RT-PCR detection of enterovirus, hepatitis A virus, Norwalk I viruses, Norwalk II viruses, astrovirus and rotavirus genomes over 12 months. All the 68 water samples tested were positive for the quantitative detection of somatic coliphages (range of concentrations: 4 x 10(2) to 1.6 x10(5) FUl(-1)). Infectious enteroviruses were isolated by a cell culture system in only two (3%) of the 68 concentrated water samples tested, whereas enterovirus genomes were detectable in 60 (88%) of the same samples. A positive RT-PCR detection of the genome of hepatitis A virus, Norwalk-like virus genogroup II, astrovirus, rotavirus and Norwalk-like virus genogroup I was demonstrated, respectively, in 1.5% (1/68), 1.5% (1/68), 3% (2/68), 0% and 0% of the 68 concentrated water samples tested. All of these four water samples were positive for the detection of enterovirus genomes, whereas only one of them was positive for the isolation of enteroviruses on cell culture. Moreover, the genomic detection of human pathogenic viruses appeared not to be statistically associated with the concentration levels of somatic coliphages in the 68 concentrated water samples tested (Wilcoxon rank test; P=0.14). Taken together, our findings indicate that the quantitative detection of somatic coliphages and the isolation of enteroviruses on cell culture are not suitable parameters for the control of the viral contamination in surface water, whereas the detection of enterovirus genomes may be useful for predicting the presence of waterborne viruses.

  7. Optimization of a Reusable Hollow-Fiber Ultrafilter for Simultaneous Concentration of Enteric Bacteria, Protozoa, and Viruses from Water

    PubMed Central

    Morales-Morales, Hugo A.; Vidal, Guadalupe; Olszewski, John; Rock, Channah M.; Dasgupta, Debanjana; Oshima, Kevin H.; Smith, Geoffrey B.

    2003-01-01

    The detection and identification of pathogens from water samples remain challenging due to variations in recovery rates and the cost of procedures. Ultrafiltration offers the possibility to concentrate viral, bacterial, and protozoan organisms in a single process by using size-exclusion-based filtration. In this study, two hollow-fiber ultrafilters with 50,000-molecular-weight cutoffs were evaluated to concentrate microorganisms from 2- and 10-liter water samples. When known quantities (105 to 106 CFU/liter) of two species of enteric bacteria were introduced and concentrated from 2 liters of sterile water, the addition of 0.1% Tween 80 increased Escherichia coli strain K-12 recoveries from 70 to 84% and Salmonella enterica serovar Enteritidis recoveries from 36 to 72%. An E. coli antibiotic-resistant strain, XL1-Blue, was recovered at a level (87%) similar to that for strain K-12 (96%) from 10 liters of sterile water. When E. coli XL1-Blue was introduced into 10 liters of nonsterile Rio Grande water with higher turbidity levels (23 to 29 nephelometric turbidity units) at two inoculum levels (9 × 105 and 2.4 × 103 per liter), the recovery efficiencies were 89 and 92%, respectively. The simultaneous addition of E. coli XL1-Blue (9 × 105 CFU/liter), Cryptosporidium parvum oocysts (10 oocysts/liter), phage T1 (105 PFU/liter), and phage PP7 (105 PFU/liter) to 10 liters of Rio Grande surface water resulted in mean recoveries of 96, 54, 59, and 46%, respectively. Using a variety of surface waters from around the United States, we obtained recovery efficiencies for bacteria and viruses that were similar to those observed with the Rio Grande samples, but recovery of Cryptosporidium oocysts was decreased, averaging 32% (the site of collection of these samples had previously been identified as problematic for oocyst recovery). Results indicate that the use of ultrafiltration for simultaneous recovery of bacterial, viral, and protozoan pathogens from variable surface waters

  8. Immunohistochemical detection and localization of new type gosling viral enteritis virus in paraformaldehyde-fixed paraffin-embedded tissue.

    PubMed

    Chen, Shun; Cheng, Anchun; Wang, Mingshu; Zhu, Dekang; Luo, Qihui; Liu, Fei; Chen, Xiaoyue

    2009-08-15

    To determine the distribution and localization of new type gosling viral enteritis virus (NGVEV) in paraformaldehyde-fixed paraffin-embedded tissues of experimentally infected goslings, for the first time, an immunohistochemical (IHC) staining method was reported. Anti-NGVEV polyclonal serum was obtained from the rabbits immunized with purified NGVEV antigen, which was extracted by caprylic-ammonium sulphate method and purified through High-Q columns anion exchange chromatography. Three-day-old NGVEV-free goslings were orally inoculated with NGVEV-CN strain suspension as infection group and phosphate buffered saline solution (PBS) as control group, respectively. The tissues were collected at sequential time points between 0.5 and 720h post inoculation (PI), and prepared for IHC staining and ultra-structural observation. The positive immunoreactivity could be readily detected in the lymphoid and gastrointestinal organs of infected goslings as early as 48 h PI, in the liver, kidney, pancreas and myocardium from 72 h, and in the cerebrum and cerebellum from 96 h, while it was hardly detected in the respiratory organs at any time. The positive staining reaction could be detected in NGVEV-infected goslings until 600 h PI, and no positive staining cell could be observed in the controls. The highest levels of viral antigen were found in the bursa of Fabricius (BF), thymus, proventriculus, gizzard and intestine tract, moreover, the liver, kidney, spleen, myocardium and pancreas were intensively and widely stained. The target cells had a ubiquitous distribution, especially included the epithelial cells, endothelial cells, superficial and crypt mucosal cells, glandular cells, fibrocytes, macrophages and lymphocytes, which served as the principal sites for antigen localization. The ultra-structural observation by transmission electron microscope (TEM) further indicated that NGVEV particles could be widely detected in the lymphoid and digestive organs of infected goslings from

  9. Real-time PCR-based infectivity assay for the titration of turkey hemorrhagic enteritis virus, an adenovirus, in live vaccines.

    PubMed

    Mahsoub, Hassan M; Evans, Nicholas P; Beach, Nathan M; Yuan, Lijuan; Zimmerman, Kurt; Pierson, Frank W

    2017-01-01

    The current in vitro titration method for turkey hemorrhagic enteritis virus (THEV) is the end-point dilution assay (EPD) in suspension cell culture (CC). This assay is subjective and results in high variability among vaccine lots. In this study, a new in vitro infectivity method combining a SYBR Green I-based qPCR assay and CC was developed for titration of live hemorrhagic enteritis (HE) CC vaccines. The qPCR was used to determine the virus genome copy number (vGCN) of the internalized virus particles following inoculation of susceptible RP19 cells with 1 vaccine label dose. The measured vGCN represents the number of infectious viral particles (IVP) per 1 dose. This method was used to compare 9 vaccine lots from 3 companies in the United States. Significant lot-to-lot variations within the same company and among the various companies were found in genomic and qPCR-based infectious titer per label dose. A positive linear relationship was found between qPCR infectious titer and genomic titer. Further, considerable variations in CCID50 titers were found among tested vaccine lots, indicating the high variability of the current titration methods. The new method provides an alternative to classical titration assays and can help reduce variation among HE vaccine products.

  10. Heparan Sulfate-Binding Foot-and-Mouth Disease Virus Enters Cells Via Caveolae-Mediated Endocytosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Foot-and-mouth disease virus (FMDV) utilizes different cell surface macromolecules to facilitate infection of cultured cells. Virus which is virulent for susceptible animals infects cells via four members of the alpha V subclass of cellular integrins. In contrast, tissue culture adaptation of some...

  11. Field-based evaluation of a male-specific (F+) RNA coliphage concentration method

    EPA Science Inventory

    Fecal contamination of water poses a significant risk to public health due to the potential presence of pathogens, including enteric viruses. Thus, sensitive, reliable and easy to use methods for the detection of microorganisms are needed to evaluate water quality. In this stud...

  12. Emergence of a New Lineage of Dengue Virus Type 2 Identified in Travelers Entering Western Australia from Indonesia, 2010-2012

    PubMed Central

    Ernst, Timo; McCarthy, Suzi; Chidlow, Glenys; Luang-Suarkia, Dagwin; Holmes, Edward C.; Smith, David W.; Imrie, Allison

    2015-01-01

    Dengue virus (DENV) transmission is ubiquitous throughout the tropics. More than 70% of the current global dengue disease burden is borne by people who live in the Asia-Pacific region. We sequenced the E gene of DENV isolated from travellers entering Western Australia between 2010–2012, most of whom visited Indonesia, and identified a diverse array of DENV1-4, including multiple co-circulating viral lineages. Most viruses were closely related to lineages known to have circulated in Indonesia for some time, indicating that this geographic region serves as a major hub for dengue genetic diversity. Most notably, we identified a new lineage of DENV-2 (Cosmopolitan genotype) that emerged in Bali in 2011–2012. The spread of this lineage should clearly be monitored. Surveillance of symptomatic returned travellers provides important and timely information on circulating DENV serotypes and genotypes, and can reveal the herald wave of dengue and other emerging infectious diseases. PMID:25635775

  13. Molecular assays for targeting human and bovine enteric viruses in coastal waters and their application for library-independent source tracking

    USGS Publications Warehouse

    Fong, T.-T.; Griffin, Dale W.; Lipp, E.K.

    2005-01-01

    Rapid population growth and urban development along waterways and coastal areas have led to decreasing water quality. To examine the effects of upstream anthropogenic activities on microbiological water quality, methods for source-specific testing are required. In this study, molecular assays targeting human enteroviruses (HEV), bovine enteroviruses (BEV), and human adenoviruses (HAdV) were developed and used to identify major sources of fecal contamination in the lower Altamaha River, Georgia. Two-liter grab samples were collected monthly from five tidally influenced stations between July and December 2002. Samples were analyzed by reverse transcription- and nested-PCR. PCR results were confirmed by dot blot hybridization. Eleven and 17 of the 30 surface water samples tested positive for HAdV and HEV, respectively. Two-thirds of the samples tested positive for either HEV or HAdV, and the viruses occurred simultaneously in 26% of samples. BEV were detected in 11 of 30 surface water samples. Binary logistic regression analysis showed that the presence of both human and bovine enteric viruses was not significantly related to either fecal coliform or total coliform levels. The presence of these viruses was directly related to dissolved oxygen and streamflow but inversely related to water temperature, rainfall in the 30 days preceding sampling, and chlorophyll-?? concentrations. The stringent host specificity of enteric viruses makes them good library-independent indicators for identification of water pollution sources. Viral pathogen detection by PCR is a highly sensitive and easy-to-use tool for rapid assessment of water quality and fecal contamination when public health risk characterization is not necessary. Copyright ?? 2005, American Society for Microbiology. All Rights Reserved.

  14. Comparison of RNA extraction kits for the purification and detection of an enteric virus surrogate on green onions via RT-PCR.

    PubMed

    Xu, Ruoyang; Shieh, Y Carol; Stewart, Diana S

    2017-01-01

    Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) offers a rapid and sensitive molecular method for detection of enteric viruses. Unfortunately, these assays are often hampered by the low virus titre found in foods and PCR inhibition due to matrix carryover during RNA extraction. Four commercial RNA extraction kits (Qiagen's QIAamp Viral RNA Mini and UltraSens Virus kits, MoBio UltraClean Tissue & Cells RNA Isolation kit, and Ambion MagMAX Viral RNA Isolation kit) were evaluated for their ability to extract and purify MS2 bacteriophage RNA, an enteric virus surrogate, from inoculated green onions, a food which has been associated with viral gastroenteritis outbreaks. Inoculated green onion wash concentrates and green onion pieces with and without Qiagen QIAshredder homogenization were assayed in the kit comparison. MS2 detection and PCR inhibition were evaluated using a duplex real-time RT-PCR for MS2 and an exogenous internal amplification control (IAC) assay. Without homogenization, MS2 inoculated at 40pfu/g was detected in at least 4 lots of green onion wash concentrates using the silica-membrane spin-column kits. Inhibition was a factor for the magnetic silica-based MagMAX kit, which resulted in detection of MS2 in 1 of 5. Addition of QIAshredder homogenization prior to extraction did not adversely affect the silica-membrane kit results but improved the MS2 detection by MagMAX to 5 of 5 lots. Use of a 1:10 dilution of primary RNA extracts also improved detection. The QIAamp Viral RNA Mini and MagMAX kits were further compared for detection of MS2 from green onion pieces inoculated at 20 and 5pfu/g. Using homogenization, the MagMAX kit detected 20pfu/g in only 1 of 2 green onion lots, whereas the QIAamp Viral RNA kit detected 2 of 2 lots at 5 pfu/g without homogenization.

  15. Molecular Assays for Targeting Human and Bovine Enteric Viruses in Coastal Waters and Their Application for Library-Independent Source Tracking

    PubMed Central

    Fong, Theng-Theng; Griffin, Dale W.; Lipp, Erin K.

    2005-01-01

    Rapid population growth and urban development along waterways and coastal areas have led to decreasing water quality. To examine the effects of upstream anthropogenic activities on microbiological water quality, methods for source-specific testing are required. In this study, molecular assays targeting human enteroviruses (HEV), bovine enteroviruses (BEV), and human adenoviruses (HAdV) were developed and used to identify major sources of fecal contamination in the lower Altamaha River, Georgia. Two-liter grab samples were collected monthly from five tidally influenced stations between July and December 2002. Samples were analyzed by reverse transcription- and nested-PCR. PCR results were confirmed by dot blot hybridization. Eleven and 17 of the 30 surface water samples tested positive for HAdV and HEV, respectively. Two-thirds of the samples tested positive for either HEV or HAdV, and the viruses occurred simultaneously in 26% of samples. BEV were detected in 11 of 30 surface water samples. Binary logistic regression analysis showed that the presence of both human and bovine enteric viruses was not significantly related to either fecal coliform or total coliform levels. The presence of these viruses was directly related to dissolved oxygen and streamflow but inversely related to water temperature, rainfall in the 30 days preceding sampling, and chlorophyll-a concentrations. The stringent host specificity of enteric viruses makes them good library-independent indicators for identification of water pollution sources. Viral pathogen detection by PCR is a highly sensitive and easy-to-use tool for rapid assessment of water quality and fecal contamination when public health risk characterization is not necessary. PMID:15812040

  16. Enterically transmitted non-A, non-B hepatitis: recovery of virus-like particles from an epidemic in south Delhi and transmission studies in rhesus monkeys.

    PubMed

    Panda, S K; Datta, R; Kaur, J; Zuckerman, A J; Nayak, N C

    1989-10-01

    An epidemic of viral hepatitis, serologically characterized as due to non-A, non-B hepatitis, occurred in a village of South Delhi, India, in December, 1986, through January, 1987. Water contaminated with fecal matter was the apparent source of infection. Disease-associated virus-like particles were detected by immune electron microscopy in the feces of three patients within 5 days of illness. The virus-like particles were agglutinated by autologous acute-phase serum but not by convalescent serum. Rhesus monkeys inoculated with particle-containing fecal suspensions developed biochemical and morphologic features of acute, self-limited hepatitis. The findings in the present study and in earlier investigations of sporadic non-A, non-B hepatitis suggest that (i) the epidemic form and a proportion of sporadic cases of this infection in India may be related, both being enterically transmitted and associated with infection by a 27- to 32-nm virus-like particle, (ii) antibody responses to this virus occur early in disease and are transient and (iii) the rhesus monkey may prove to be a suitable model for studies of epidemic non-A, non-B hepatitis.

  17. Solar water disinfection (SODIS) of Escherichia coli, Enterococcus spp., and MS2 coliphage: effects of additives and alternative container materials.

    PubMed

    Fisher, Michael B; Iriarte, Mercedes; Nelson, Kara L

    2012-04-15

    The use of alternative container materials and added oxidants accelerated the inactivation of MS2 coliphage and Escherichia coli and Enterococcus spp. bacteria during solar water disinfection (SODIS) trials. Specifically, bottles made from polypropylene copolymer (PPCO), a partially UVB-transparent plastic, resulted in three-log inactivation of these organisms in approximately half the time required for disinfection in bottles made from PET, polycarbonate, or Tritan(®), which absorb most UVB light. Furthermore, the addition of 125 mg/L sodium percarbonate in combination with either citric acid or copper plus ascorbate tended to accelerate inactivation by factors of 1.4-19. Finally, it was observed that the inactivation of E. coli and enterococci derived from local wastewater was far slower than the inactivation of laboratory-cultured E. coli and Enterococcus spp., while the inactivation of MS2 was slowest of all. These results highlight the importance of UVB in SODIS under certain conditions, and also the greater sunlight resistance of some viruses and of bacteria of fecal origin, as compared to the laboratory-cultured bacteria commonly used to model their inactivation. Furthermore, this study illustrates promising new avenues for accelerating the inactivation of bacteria and viruses by solar disinfection.

  18. Methods for recovery of hepatitis A virus (HAV) and other viruses from processed foods and detection of HAV by nested RT-PCR and TaqMan RT-PCR.

    PubMed

    Love, David C; Casteel, Michael J; Meschke, John S; Sobsey, Mark D

    2008-08-15

    Enteric viruses are important agents of foodborne disease. Unfortunately, robust, quantitative methods for sampling and analysis of enteric and other viruses in processed or complex foods are not well-established. As a result, epidemiologically determined etiologies or pathogen sources in foodborne outbreaks are rarely confirmed by virological analysis. In this study, an acid-adsorption elution concentration (AEC) method previously used to monitor virus occurrence and investigate enteric virus outbreaks in shellfish was adapted for examination of processed food items, namely tomato sauce and blended strawberries. Hepatitis A virus (HAV), poliovirus, and coliphage MS2 (MS2) were seeded in 10 or 30 g samples of tomato sauce or blended strawberries, recovered by AEC, and quantified by cell culture infectivity assay. In addition, nested reverse transcription-polymerase chain reaction (RT-PCR) and TaqMan RT-PCR assays were used to detect HAV RNA. Viruses were efficiently adsorbed to foods as an initial concentration step, with infectious HAV and MS2 adsorption of 67% and 93%, respectively, to tomato sauce, and 89% and 99%, respectively, to blended strawberries. Forty-three to 65% of HAV and poliovirus were subsequently eluted and recovered from tomato sauce using 0.5 M threonine, pH 7.2. The lower limits of HAV detection were at initial seeding levels of 14 PFU/g of tomato sauce and 33 PFU/g of blended strawberries. Unlike TaqMan RT-PCR, nested RT-PCR was not inhibited by undiluted final RNA extracts of tomato sauce or blended strawberries. The successful adaptation of the AEC method for enteric and other virus recovery, quantitation and detection in processed foods demonstrates its potential for use in the investigation of foodborne outbreaks of viral etiology and for validation of virus disinfection and sanitary processing procedures used by the food industry.

  19. Enteric virus status of turkey flocks over time: molecular diagnostic studies beginning on the day of placement.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Poultry enteric disease is often associated with numerous viral and/or bacterial infections, including avian reoviruses, rotaviruses, astroviruses, parvoviruses, and Escherichia coli. These potential etiologic agents are often present in combination in a flock or individual birds, but in general it ...

  20. Properties of the Deoxyribonucleic Acid Contained in the Defective Particle Coliphage 15 1

    PubMed Central

    Frampton, E. W.; Mandel, M.

    1970-01-01

    Escherichia coli strain 15 TAU, which requires thymine, arginine, and uracil for growth and harbors an apparently defective prophage, was induced by exposure to ultraviolet light (580 ergs/mm2) or to mitomycin C (5 μg/ml). Phage particles (coliphage 15) were recovered from the resulting lysate by treatment with deoxyribonuclease, filtration, and several cycles of differential centrifugation. Analysis of the phage particles obtained by using cesium chloride density gradient centrifugation in a preparative ultracentrifuge resulted in the resolution of three components. The major component had a peak density of 1.52 to 1.53 g/cm3 followed by components with densities of 1.5 and 1.49 g/cm3. The guanine plus cytosine content of coliphage 15 deoxyribonucleic acid (DNA) was determined by both analytical ultracentrifugation in cesium chloride and by thermal denaturation in standard saline citrate buffer. Respective values of 46.4 ± 1% and 46.6 ± 1% guanine plus cytosine content were obtained. Coliphage 15 DNA formed molecular hybrids with messenger ribonucleic acid (RNA) from both uninduced and ultraviolet-induced cultures of E. coli 15 TAU, but did not hybridize with E. coli ribosomal RNA. The molecular weight of coliphage 15 DNA was determined by constant velocity sucrose density gradient centrifugation to be about 33 × 106 daltons. PMID:4909911

  1. Transport of enterococci and F+ coliphage through the saturated zone of the beach aquifer.

    PubMed

    de Sieyes, Nicholas R; Russell, Todd L; Brown, Kendra I; Mohanty, Sanjay K; Boehm, Alexandria B

    2016-02-01

    Coastal groundwater has been implicated as a source of microbial pollution to recreational beaches. However, there is little work investigating the transport of fecal microbes through beach aquifers where waters of variable salinity are present. In this study, the potential for fecal indicator organisms enterococci (ENT) and F+ coliphage to be transported through marine beach aquifers was investigated. Native sediment and groundwaters were collected from the fresh and saline sections of the subterranean estuary at three beaches along the California coast where coastal communities utilize septic systems for wastewater treatment. Groundwaters were seeded with sewage and removal of F+ coliphage and ENT by the sediments during saturated flow was tested in laboratory column experiments. Removal varied significantly between beach and organism. F+ coliphage was removed to a greater extent than ENT, and removal was greater in saline sediments and groundwater than fresh. At one of the three beaches, a field experiment was conducted to investigate the attenuation of F+ coliphage and ENT down gradient of a septic leach field. ENT were detected up to 24 m from the leach field. The column study and field observations together suggest ENT can be mobile within native aquifer sediments and groundwater under certain conditions.

  2. DETECTION OF FRNA COLIPHAGES IN GROUNDWATER: INTERFERENCE WITH THE ASSAY BY SOMATIC SALMONELLA BACTERIOPHAGES

    EPA Science Inventory

    Groundwater samples from two sites in Alabama, USA were plaque assayed for F-specific RNA (FRNA) coliphages using Salmonella typhimurium WG49 as the host bacterium. While numerous plaques were detected with WG49 (a strain possessing Escherichia coli F pili), plaques were also obs...

  3. Molecular and serological surveillance of canine enteric viruses in stray dogs from Vila do Maio, Cape Verde

    PubMed Central

    2014-01-01

    Background Infections caused by canine parvovirus, canine distemper virus and canine coronavirus are an important cause of mortality and morbidity in dogs worldwide. Prior to this study, no information was available concerning the incidence and prevalence of these viruses in Cape Verde archipelago. Results To provide information regarding the health status of the canine population in Vila do Maio, Maio Island, Cape Verde, 53 rectal swabs were collected from 53 stray dogs during 2010 and 93 rectal swabs and 88 blood samples were collected from 125 stray dogs in 2011. All rectal swabs (2010 n = 53; 2011 n = 93) were analysed for the presence of canine parvovirus, canine distemper virus and canine coronavirus nucleic acids by quantitative PCR methods. Specific antibodies against canine distemper virus and canine parvovirus were also assessed (2011 n = 88). From the 2010 sampling, 43.3% (23/53) were positive for canine parvovirus DNA, 11.3% (6/53) for canine distemper virus RNA and 1.9% (1/53) for canine coronavirus RNA. In 2011, the prevalence values for canine parvovirus and canine coronavirus were quite similar to those from the previous year, respectively 44.1% (41/93), and 1.1% (1/93), but canine distemper virus was not detected in any of the samples analysed (0%, 0/93). Antibodies against canine parvovirus were detected in 71.6% (63/88) blood samples and the seroprevalence found for canine distemper virus was 51.1% (45/88). Conclusions This study discloses the data obtained in a molecular and serological epidemiological surveillance carried out in urban populations of stray and domestic animals. Virus transmission and spreading occurs easily in large dog populations leading to high mortality rates particularly in unvaccinated susceptible animals. In addition, these animals can act as disease reservoirs for wild animal populations by occasional contact. Identification of susceptible wildlife of Maio Island is of upmost importance to evaluate the risk

  4. Different Behavior of Enteric Bacteria and Viruses in Clay and Sandy Soils after Biofertilization with Swine Digestate.

    PubMed

    Fongaro, Gislaine; García-González, María C; Hernández, Marta; Kunz, Airton; Barardi, Célia R M; Rodríguez-Lázaro, David

    2017-01-01

    Enteric pathogens from biofertilizer can accumulate in the soil, subsequently contaminating water and crops. We evaluated the survival, percolation and leaching of model enteric pathogens in clay and sandy soils after biofertilization with swine digestate: PhiX-174, mengovirus (vMC0), Salmonella enterica Typhimurium and Escherichia coli O157:H7 were used as biomarkers. The survival of vMC0 and PhiX-174 in clay soil was significantly lower than in sandy soil (iT90 values of 10.520 ± 0.600 vs. 21.270 ± 1.100 and 12.040 ± 0.010 vs. 43.470 ± 1.300, respectively) and PhiX-174 showed faster percolation and leaching in sandy soil than clay soil (iT90 values of 0.46 and 2.43, respectively). S. enterica Typhimurium was percolated and inactivated more slowly than E. coli O157:H7 (iT90 values of 9.340 ± 0.200 vs. 6.620 ± 0.500 and 11.900 ± 0.900 vs. 10.750 ± 0.900 in clay and sandy soils, respectively), such that E. coli O157:H7 was transferred more quickly to the deeper layers of both soils evaluated (percolation). Our findings suggest that E. coli O157:H7 may serve as a useful microbial biomarker of depth contamination and leaching in clay and sandy soil and that bacteriophage could be used as an indicator of enteric pathogen persistence. Our study contributes to development of predictive models for enteric pathogen behavior in soils, and for potential water and food contamination associated with biofertilization, useful for risk management and mitigation in swine digestate recycling.

  5. Different Behavior of Enteric Bacteria and Viruses in Clay and Sandy Soils after Biofertilization with Swine Digestate

    PubMed Central

    Fongaro, Gislaine; García-González, María C.; Hernández, Marta; Kunz, Airton; Barardi, Célia R. M.; Rodríguez-Lázaro, David

    2017-01-01

    Enteric pathogens from biofertilizer can accumulate in the soil, subsequently contaminating water and crops. We evaluated the survival, percolation and leaching of model enteric pathogens in clay and sandy soils after biofertilization with swine digestate: PhiX-174, mengovirus (vMC0), Salmonella enterica Typhimurium and Escherichia coli O157:H7 were used as biomarkers. The survival of vMC0 and PhiX-174 in clay soil was significantly lower than in sandy soil (iT90 values of 10.520 ± 0.600 vs. 21.270 ± 1.100 and 12.040 ± 0.010 vs. 43.470 ± 1.300, respectively) and PhiX-174 showed faster percolation and leaching in sandy soil than clay soil (iT90 values of 0.46 and 2.43, respectively). S. enterica Typhimurium was percolated and inactivated more slowly than E. coli O157:H7 (iT90 values of 9.340 ± 0.200 vs. 6.620 ± 0.500 and 11.900 ± 0.900 vs. 10.750 ± 0.900 in clay and sandy soils, respectively), such that E. coli O157:H7 was transferred more quickly to the deeper layers of both soils evaluated (percolation). Our findings suggest that E. coli O157:H7 may serve as a useful microbial biomarker of depth contamination and leaching in clay and sandy soil and that bacteriophage could be used as an indicator of enteric pathogen persistence. Our study contributes to development of predictive models for enteric pathogen behavior in soils, and for potential water and food contamination associated with biofertilization, useful for risk management and mitigation in swine digestate recycling. PMID:28197137

  6. Enteric Viruses Ameliorate Gut Inflammation via Toll-like Receptor 3 and Toll-like Receptor 7-Mediated Interferon-β Production.

    PubMed

    Yang, Jin-Young; Kim, Min-Soo; Kim, Eugene; Cheon, Jae Hee; Lee, Yong-Soo; Kim, Yeji; Lee, Su-Hyun; Seo, Sang-Uk; Shin, Seung-Ho; Choi, Sun Shim; Kim, Bumseok; Chang, Sun-Young; Ko, Hyun-Jeong; Bae, Jin-Woo; Kweon, Mi-Na

    2016-04-19

    Metagenomic studies show that diverse resident viruses inhabit the healthy gut; however, little is known about the role of these viruses in the maintenance of gut homeostasis. We found that mice treated with antiviral cocktail displayed more severe dextran sulfate sodium (DSS)-induced colitis compared with untreated mice. DSS-induced colitis was associated with altered enteric viral abundance and composition. When wild-type mice were reconstituted with Toll-like receptor 3 (TLR3) or TLR7 agonists or inactivated rotavirus, colitis symptoms were significantly ameliorated. Mice deficient in both TLR3 and TLR7 were more susceptible to DSS-induced experimental colitis. In humans, combined TLR3 and TLR7 genetic variations significantly influenced the severity of ulcerative colitis. Plasmacytoid dendritic cells isolated from inflamed mouse colon produced interferon-β in a TLR3 and TLR7-dependent manner. These results imply that recognition of resident viruses by TLR3 and TLR7 is required for protective immunity during gut inflammation.

  7. Farm-level prevalence and risk factors for detection of hepatitis E virus, porcine enteric calicivirus, and rotavirus in Canadian finisher pigs.

    PubMed

    Wilhelm, Barbara; Leblanc, Danielle; Leger, David; Gow, Sheryl; Deckert, Anne; Pearl, David L; Friendship, Robert; Rajić, Andrijana; Houde, Alain; McEwen, Scott

    2016-04-01

    Hepatitis E virus (HEV), norovirus (NoV), and rotavirus (RV) are all hypothesized to infect humans zoonotically via exposure through swine and pork. Our study objectives were to estimate Canadian farm-level prevalence of HEV, NoV [specifically porcine enteric calicivirus (PEC)], and RV in finisher pigs, and to study risk factors for farm level viral detection. Farms were recruited using the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS) and FoodNet Canada on-farm sampling platforms. Six pooled groups of fecal samples were collected from participating farms, and a questionnaire capturing farm management and biosecurity practices was completed. Samples were assayed using validated real-time polymerase chain reaction (RT-PCR). We modeled predictors for farm level viral RNA detection using logistic and exact logistic regression. Seventy-two herds were sampled: 51 CIPARS herds (15 sampled twice) and 21 FoodNet Canada herds (one sampled twice). Hepatitis E virus was detected in 30/88 farms [34.1% (95% CI 25.0%, 44.5%)]; PEC in 18 [20.5% (95% CI: 13.4%, 30.0%)], and RV in 6 farms [6.8% (95% CI: 3.2%, 14.1%)]. Farm-level prevalence of viruses varied with province and sampling platform. Requiring shower-in and providing boots for visitors were significant predictors (P < 0.05) in single fixed effect mixed logistic regression analysis for detection of HEV and PEC, respectively. In contrast, all RV positive farms provided boots and coveralls, and 5 of 6 farms required shower-in. We hypothesized that these biosecurity measures delayed the mean age of RV infection, resulting in an association with RV detection in finishers. Obtaining feeder pigs from multiple sources was consistently associated with greater odds of detecting each virus.

  8. Cellular microRNA miR-181b inhibits replication of mink enteritis virus by repression of non-structural protein 1 translation.

    PubMed

    Sun, Jia-zeng; Wang, Jigui; Yuan, Daoli; Wang, Shuang; Li, Zhili; Yi, Bao; Mao, Yaping; Hou, Qiang; Liu, Weiquan

    2013-01-01

    Mink enteritis virus (MEV) is one of the most important viral pathogens in the mink industry. Recent studies have showed that microRNAs (miRNAs), small noncoding RNAs of length ranging from 18-23 nucleotides (nt) participate in host-pathogen interaction networks; however, whether or not miRNAs are involved in MEV infection has not been reported. Our study revealed that miRNA miR-181b inhibited replication of MEV in the feline kidney (F81) cell line by targeting the MEV non-structural protein 1 (NS1) messenger RNA (mRNA) coding region, resulting in NS1 translational repression, while MEV infection reduced miR-181b expression. This is the first description of cellular miRNAs modulating MEV infection in F81 cells, providing further insight into the mechanisms of viral infection, and may be useful in development of naturally-occurring miRNAs antiviral strategies.

  9. Cellular microRNA miR-181b Inhibits Replication of Mink Enteritis Virus by Repression of Non-Structural Protein 1 Translation

    PubMed Central

    Sun, Jia-zeng; Wang, Jigui; Yuan, Daoli; Wang, Shuang; Li, Zhili; Yi, Bao; Mao, Yaping; Hou, Qiang; Liu, Weiquan

    2013-01-01

    Mink enteritis virus (MEV) is one of the most important viral pathogens in the mink industry. Recent studies have showed that microRNAs (miRNAs), small noncoding RNAs of length ranging from 18–23 nucleotides (nt) participate in host-pathogen interaction networks; however, whether or not miRNAs are involved in MEV infection has not been reported. Our study revealed that miRNA miR-181b inhibited replication of MEV in the feline kidney (F81) cell line by targeting the MEV non-structural protein 1 (NS1) messenger RNA (mRNA) coding region, resulting in NS1 translational repression, while MEV infection reduced miR-181b expression. This is the first description of cellular miRNAs modulating MEV infection in F81 cells, providing further insight into the mechanisms of viral infection, and may be useful in development of naturally-occurring miRNAs antiviral strategies. PMID:24349084

  10. Histopathology, immunohistochemistry, in situ apoptosis, and ultrastructure characterization of the digestive and lymphoid organs of new type gosling viral enteritis virus experimentally infected gosling.

    PubMed

    Chen, S; Cheng, A C; Wang, M S; Zhu, D K; Jia, R Y; Luo, Q H; Cui, H M; Zhou, Y; Wang, Y; Xu, Z W; Chen, Z L; Chen, X Y; Wang, X Y

    2010-04-01

    Here, for the first time, to colocalize new type gosling viral enteritis virus (NGVEV) with histological lesions and in situ apoptosis in the digestive organs (esophagus, proventriculus, gizzard, small intestine, cecum, rectum, liver, and pancreas) and the lymphoid organs (bursa of Fabricius, thymus, Harderian gland, and spleen) of experimentally infected goslings, portions of tissues were collected at sequential infection time points and examined by histopathology for histological lesions, immunohistochemical staining for viral antigens, ultrastructural observation by transmission electron microscope (TEM) for virus particles and apoptotic cells, and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling assay for in situ apoptosis. The hyperemia, hemorrhage, infiltration of lymphocytes, progressive lymphoid depletion, apoptosis, and necrosis were readily observed in the lymphoid organs and intestine tract by histopathological examination. The NGVEV particles and viral antigens widely appeared in the small intestine and bursa of Fabricius as early as 2 d postinfection (PI) by TEM and immunohistochemical staining, and the presence and quantity of it reached a maximum during 6 to 12 d PI. The principal sites for NGVEV were endothelial cells, epithelia, mucosal cells, glandular cells, fibrocytes, macrophages, and lymphocytes. A series of apoptotic morphological changes including chromatin condensation and margination, cytoplasmic shrinkage, and formation of apoptotic body were observed by TEM, and the number of apoptotic cells was largely increased from 4 d PI and peaked at 9 d PI by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling analysis. The histological organ lesions and apoptosis in vivo were generally associated with sites of NGVEV localization, which can be regarded as the cause of death. This work may shed light on the pathogenesis of new type gosling viral enteritis and put new

  11. Enteral feedings.

    PubMed

    Chernoff, R

    1980-01-01

    The benefits, equipment used, commercially available sources, and the indications and techniques for administration of enteral nutrients are reviewed. In many malabsorption states, enteral feeding is preferable and parenteral nutrients are seldom indicated. Transitional enteral nutrient support usually is indicated after parenteral nutrient therapy. Enteral tube-feeding formulas should be matched to the patient's needs; formulas using blenderized natural foods or intact isolated nutrients are appropriate for patients with intact gastrointestinal tracts. Patients should be monitored for glucosuria and hyperglycemia, bloating, nausea, dehydration, and renal, hepatic and hematologic status. Formula dilution, and a reduced flow rate or use of continuous-drip feeding, will reduce the incidence of osmotic diarrhea. The effectiveness, low cost and low potential for serious complications make enteral feeding preferable to parenteral nutrient therapy for many patients.

  12. Validation of Internal Controls for Extraction and Amplification of Nucleic Acids from Enteric Viruses in Water Samples ▿ †

    PubMed Central

    Hata, Akihiko; Katayama, Hiroyuki; Kitajima, Masaaki; Visvanathan, Chettiyappan; Nol, Chea; Furumai, Hiroaki

    2011-01-01

    Inhibitors that reduce viral nucleic acid extraction efficiency and interfere with cDNA synthesis and/or polymerase activity affect the molecular detection of viruses in aquatic environments. To overcome these significant problems, we developed a methodology for assessing nucleic acid yields and DNA amplification efficiencies for environmental water samples. This involved adding particles of adenovirus type 5 and murine norovirus and newly developed primer-sharing controls, which are amplified with the same primer pairs and result in the same amplicon sizes as the targets, to these samples. We found that nucleic acid loss during the extraction process, rather than reverse transcription-PCR (RT-PCR) inhibition, more significantly attributed to underestimation of the presence of viral genomes in the environmental water samples tested in this study. Our success rate for satisfactorily amplifying viral RNAs and DNAs by RT-PCR was higher than that for obtaining adequate nucleic acid preparations. We found that inhibitory properties were greatest when we used larger sample volumes. A magnetic silica bead-based RNA extraction method effectively removed inhibitors that interfere with viral nucleic acid extraction and RT-PCR. To our knowledge, this is the first study to assess the inhibitory properties of environmental water samples by using both control virus particles and primer-sharing controls. PMID:21602369

  13. Prevalence of antibodies to bluetongue virus and Anaplasma marginale in Montana yearling cattle entering Alberta feedlots: Fall 2001.

    PubMed

    Van Donkersgoed, Joyce; Gertonson, Arnold; Bridges, Marc; Raths, Dick; Dargatz, David; Wagner, Bruce; Boughton, Alice; Knoop, Doug; Walton, Thomas E

    2004-06-01

    A serologic survey was conducted in yearling cattle imported into Alberta feedlots from Montana during October 2001 to estimate the prevalence of antibodies to bluetongue virus (BTV) and Anaplasma marginale in Montana yearling cattle. The apparent prevalence of antibodies to BTV when the competitive enzyme-linked immunosorbent assay (cELISA) was used was 0.37% (21/5608). Test positive cELISA samples were also all positive when tested by virus neutralization (VN) and they reacted to 1 or more BTV serotypes, including 2, 10, 11, 13, and 17. The apparent prevalence of antibodies to A. marginale when a recombinant cELISA (rcELISA) was used with a positive cutoff at 30% inhibition was 1.93% (108/5608). When the rcELISA positive cutoff was at 42% inhibition, the apparent prevalence was 0.73% (41/5608). After the reported sensitivity and specificity of the test had been accounted for, the A. marginale antibody results were consistent with a population that was either free of exposure or had a very low prevalence for A. marginale.

  14. From Lab to Lake – Evaluation of Current Molecular Methods for the Detection of Infectious Enteric Viruses in Complex Water Matrices in an Urban Area

    PubMed Central

    Hamza, Ibrahim Ahmed; Krieger, Marion; Wilhelm, Michael; Mackowiak, Martin; Jurzik, Lars

    2016-01-01

    Quantitative PCR methods are commonly used to monitor enteric viruses in the aquatic environment because of their high sensitivity, short reaction times and relatively low operational cost. However, conclusions for public health drawn from results of such molecular techniques are limited due to their inability to determine viral infectivity. Ethidium monoazide (EMA) and propidium monoazide (PMA) are capable to penetrate the damaged or compromised capsid of the inactivated viruses and bind to the viral nucleic acids. We assessed whether dye treatment is a suitable approach to improve the ability of qPCR to distinguish between infectious and non-infectious human adenovirus, enterovirus and rotavirus A in surface water of an urban river and sewage before and after UV disinfection. Like the gold standard of cell culture assays, pretreatment EMA-/PMA-qPCR succeeded in removing false positive results which would lead to an overestimation of the viral load if only qPCR of the environmental samples was considered. A dye pretreatment could therefore provide a rapid and relatively inexpensive tool to improve the efficacy of molecular quantification methods in regards to viral infectivity. PMID:27880820

  15. Lactic acid bacteria efficiently protect human and animal intestinal epithelial and immune cells from enteric virus infection.

    PubMed

    Maragkoudakis, Petros A; Chingwaru, Walter; Gradisnik, Lidija; Tsakalidou, Effie; Cencic, Avrelija

    2010-07-31

    This study aimed to examine the potential antiviral activity of lactic acid bacteria (LAB) using animal and human intestinal and macrophage cell line models of non tumor origin. To this end, LAB strains selected on the basis of previous in vitro trials were co-incubated with cell line monolayers, which were subsequently challenged with rotavirus (RV) and transmissible gastroenteritis virus (TGEV). In order to elucidate the possible mechanism responsible for the antiviral activity, the induction of reactive oxygen species (ROS) release as well as the attachment ability of LAB on the cell lines was investigated. Various strains were found to exhibit moderate to complete monolayer protection against viral RV or TGEV disruption. Highest protection effects were recorded with the known probiotics Lactobacillus rhamnosus GG and Lactobacillus casei Shirota against both RV and TGEV, while notable antiviral activity was also attributed to Enterococcus faecium PCK38, Lactobacillus fermentum ACA-DC179, Lactobacillus pentosus PCA227 and Lactobacillus plantarum PCA236 and PCS22, depending on the cell line and virus combination used. A variable increase (of up to 50%) on the release of NO(-) and H(2)O(2) (ROS) was obtained when LAB strains were co-incubated with the cell lines, but the results were found to be LAB strain and cell line specific, apart from a small number of strains which were able to induce strong ROS release in more than one cell line. In contrast, the ability of the examined LAB strains to attach to the cell line monolayers was LAB strain but not cell line specific. Highest attachment ability was observed with L. plantarum ACA-DC 146, L. paracasei subsp. tolerans ACA-DC 4037 and E. faecium PCD71. Clear indications on the nature of the antiviral effect were evident only in the case of the L. casei Shirota against TGEV and with L. plantarum PCA236 against both RV and TGEV. In the rest of the cases, each interaction was LAB-cell line-virus specific, barring general

  16. UV induction of coliphage 186: Prophage induction as an SOS function

    SciTech Connect

    Lamont, I.; Brumby, A.M.; Egan, J.B.

    1989-07-01

    Our results show that UV induction of the 186 prophage depends upon the phage function Tum, with the mutant phenotype of turbid plaques on mitomycin plates and the expression of which is controlled by the host LexA protein. Tum function, encoded near the right-hand end of the coliphage 186 chromosome, is under the control of promoter p95. This promoter is overlapped by a sequence closely related to the consensus sequence of the LexA-binding site. It is proposed that inactivation of LexA after UV irradiation (or by genetic means) leads to prophage induction by permitting expression of Tum which, by unknown means, induces prophage. This mechanism is basically different from that seen with the UV-inducible lambdoid coliphages, which are not regulated by LexA.

  17. Alternative Fecal Indicators and Their Empirical Relationships with Enteric Viruses, Salmonella enterica, and Pseudomonas aeruginosa in Surface Waters of a Tropical Urban Catchment

    PubMed Central

    Liang, L.; Goh, S. G.; Vergara, G. G. R. V.; Fang, H. M.; Rezaeinejad, S.; Chang, S. Y.; Bayen, S.; Lee, W. A.; Sobsey, M. D.; Rose, J. B.

    2014-01-01

    The suitability of traditional microbial indicators (i.e., Escherichia coli and enterococci) has been challenged due to the lack of correlation with pathogens and evidence of possible regrowth in the natural environment. In this study, the relationships between alternative microbial indicators of potential human fecal contamination (Bacteroides thetaiotaomicron, Methanobrevibacter smithii, human polyomaviruses [HPyVs], and F+ and somatic coliphages) and pathogens (Salmonella spp., Pseudomonas aeruginosa, rotavirus, astrovirus, norovirus GI, norovirus GII, and adenovirus) were compared with those of traditional microbial indicators, as well as environmental parameters (temperature, conductivity, salinity, pH, dissolved oxygen, total organic carbon, total suspended solids, turbidity, total nitrogen, and total phosphorus). Water samples were collected from surface waters of urban catchments in Singapore. Salmonella and P. aeruginosa had significant positive correlations with most of the microbial indicators, especially E. coli and enterococci. Norovirus GII showed moderately strong positive correlations with most of the microbial indicators, except for HPyVs and coliphages. In general, high geometric means and significant correlations between human-specific markers and pathogens suggest the possibility of sewage contamination in some areas. The simultaneous detection of human-specific markers (i.e., B. thetaiotaomicron, M. smithii, and HPyVs) with E. coli and enterococcus supports the likelihood of recent fecal contamination, since the human-specific markers are unable to regrow in natural surface waters. Multiple-linear-regression results further confirm that the inclusion of M. smithii and HPyVs, together with traditional indicators, would better predict the occurrence of pathogens. Further study is needed to determine the applicability of such models to different geographical locations and environmental conditions. PMID:25416765

  18. Efficacy of Cinnamaldehyde Against Enteric Viruses and Its Activity After Incorporation Into Biodegradable Multilayer Systems of Interest in Food Packaging.

    PubMed

    Fabra, M J; Castro-Mayorga, J L; Randazzo, W; Lagarón, J M; López-Rubio, A; Aznar, R; Sánchez, G

    2016-06-01

    Cinnamaldehyde (CNMA), an organic compound that gives cinnamon its flavor and odor, was investigated for its virucidal activity on norovirus surrogates, murine norovirus (MNV) and feline calicivirus (FCV), and hepatitis A virus (HAV). Initially, different concentrations of CNMA (0.1, 0.5 and 1 %) were individually mixed with each virus at titers of ca. 6-7 log10 TCID50/ml and incubated 2 h at 4 and 37 °C. CNMA was effective in reducing the titers of norovirus surrogates in a dose-dependent manner after 2 h at 37 °C, while HAV titers were reduced by 1 log10 after treatment with 1 % of CNMA. When incubation time was extended, HAV titers were reduced by 3.4 and 2.7 log10 after overnight incubation at 37 °C with 1 and 0.5 % of CNMA, respectively. Moreover, this paper analyzed, for the first time, the antiviral activity of adding an active electrospun interlayer based on zein and CNMA to a polyhydroxybutyrate packaging material (PHB) in a multilayer form. Biodegradable multilayer systems prepared with 2.60 mg/cm(2) (~9.7 %) of CNMA completely inactivated FCV according to ISO 22196:2011, while MNV titers were reduced by 2.75 log10. When the developed multilayer films were evaluated after one month of preparation or at 25 °C, the antiviral activity was reduced as compared to freshly prepared multilayer films evaluated at 37 °C. The results show the excellent potential of this system for food contact applications as well as for active packaging technologies in order to maintain or extend food quality and safety.

  19. ECHO virus

    MedlinePlus

    ... page: //medlineplus.gov/ency/article/001340.htm ECHO virus To use the sharing features on this page, please enable JavaScript. Enteric cytopathic human orphan (ECHO) viruses are a group of viruses that lead to ...

  20. Enteric Campylobacteria and RNA Viruses Associated with Healthy and Diarrheic Humans in the Chinook Health Region of Southwestern Alberta, Canada ▿

    PubMed Central

    Inglis, G. Douglas; Boras, Valerie F.; Houde, Alain

    2011-01-01

    The presence of Campylobacter species and enteric RNA viruses in stools from diarrheic (n = 442) and healthy (n = 58) humans living in southwestern Alberta was examined (May to October 2005). A large number of diarrheic individuals who were culture negative for C. jejuni (n = 54) or C. coli (n = 19) were PCR positive for these taxa. Overall detection rates for C. jejuni and C. coli in diarrheic stools were 29% and 5%, respectively. In contrast, 3% and 0% of stools from healthy humans were positive for these taxa, respectively. Infection with C. jejuni was endemic over the study period. However, there was no difference in infection rates between individuals living in urban or rural locations. Stools from a large number of diarrheic (74%) and healthy (88%) individuals were positive for Campylobacter DNA. The prevalence rates of C. concisus, C. curvus, C. fetus, C. gracilis, C. helveticus, C. hominis, C. hyointestinalis, C. mucosalis, C. showae, C. sputorum, and C. upsaliensis DNA were either not significantly different or were significantly lower in stools from diarrheic than from healthy individuals. No C. lanienae or C. lari DNA was detected. Stools from 4% and 0% of diarrheic and healthy humans, respectively, were positive for rotavirus, sapovirus, or norovirus (GI/GII). Our results showed a high prevalence of diarrheic individuals living in southwestern Alberta who were infected by C. jejuni and, to a lesser extent, by C. coli. However, other Campylobacter species, norovirus, rotavirus, sapovirus, and bovine enteric calicivirus were either inconsequential pathogens during the study period or are not pathogens at all. PMID:21106791

  1. Complete genome sequence of virulent duck enteritis virus (DEV) strain 2085 and comparison with genome sequences of virulent and attenuated DEV strains.

    PubMed

    Wang, Jichun; Höper, Dirk; Beer, Martin; Osterrieder, Nikolaus

    2011-09-01

    We here report the complete genome sequence of the duck enteritis virus (DEV) wild-type strain 2085, an avian herpesvirus (GenBank ID: JF999965). The nucleotide sequence was derived from the 2085 genome cloned as an infectious bacterial artificial chromosome (BAC) clone. The DEV 2085 genome is 160,649-bp in length and encodes 78 predicted open reading frames (ORFs), a number identical to that identified for the attenuated DEV VAC strain (GenBank ID: EU082088.2). Comparison of the genome sequences DEV 2085 and VAC with partial sequences of the virulent CHv strain and the attenuated strain Clone-03 was carried out to identify nucleotide or amino acid polymorphisms that potentially contribute to DEV virulence. No amino acid changes were identified in 24 of the 78 ORFs, a result indicating high conservation in DEV independently of strain origin or virulence. In addition, 39 ORFs contain non-synonymous nucleotide substitutions, while 15 ORFs had nucleotide insertions or deletions, frame-shift mutations and/or non-synonymous nucleotide substitutions with an effect on ORF initiation or termination. In 7 of the 15 ORFs with high and 27 of the 39 ORFs with low variability, polymorphisms were exclusively found in DEV 2085, a finding that likely is a result of a different origin of 2085 (Europe) or VAC, Clone-03 and CHv (Eastern Asia). Five ORFs (UL2, UL12, US10, UL47 and UL41) with polymorphisms were identical between the virulent DEV 2085 and CHv but different from VAC or Clone-03. They, individually or in combination, may therefore represent DEV virulence factors. Our comparative analysis of four DEV sequences provides a comprehensive overview of DEV genome structure and identifies ORFs that are changed during serial virus passage.

  2. Distribution of coliphages against four e. Coli virotypes in hospital originated sewage sample and a sewage treatment plant in bangladesh.

    PubMed

    Alam, Muntasir; Farzana, Tasmia; Ahsan, Chowdhury Rafiqul; Yasmin, Mahmuda; Nessa, Jamalun

    2011-06-01

    The distribution of coliphages infecting different Escherichia coli virotypes (EHEC, EIEC, EPEC, ETEC) and an avirulent strain (K-12) in sewage system of a hospital and a sewage treatment plant (STP) was investigated by culture-based agar overlay methods. Coliphages were found in all the samples except stool dumping site in the sewage system of the hospital and lagoon of the STP. Bacteriophage count (pfu/ml) infecting E. coli strains showed the following ascending pattern (EHEC < EIEC < EPEC < ETEC < E coli K-12) in all the collected samples except one. Phages capable of infecting avirulent E. coli K-12 strains were present in the highest number among all the examined locations. Phages specific for E. coli K-12 presented high diversity in plaque size on the bacterial lawn. Virulent E. coli specific coliphages rarely produced plaques with diameter of 1-2 mm or over. Conventional agar overlay method was found to be not satisfactory for phage community analysis from primary stool dumping site of the hospital, probably due to the presence of high concentration of antimicrobial substances. The gradual decrease seen in the five groups of coliphage quantity with the ongoing treatment process and then the absolute absence of coliphages in the outlet of the examined treatment plant is indicative of the usefulness of the treatment processes practiced there.

  3. Development and evaluation of an immunochromatographic strip test based on the recombinant UL51 protein for detecting antibody against duck enteritis virus

    PubMed Central

    2010-01-01

    Background Duck enteritis virus (DEV) infection causes substantial economic losses to the worldwide duck-producing areas. The monitoring of DEV-specific antibodies is a key to evaluate the effect of DEV vaccine and develop rational immunization programs. Thus, in this study, an immunochromatographic strip (ICS) test was developed for detecting DEV serum antibodies. Results The ICS test is based on membrane chromatography, and uses both the purified recombinant UL51 protein conjugated with colloidal gold and goat anti-rabbit IgG conjugated with colloidal gold as tracers, the purified recombinant UL51 protein as the capture reagent at the test line, and rabbit IgG as the capture reagent at the control line. The specificity of the ICS was evaluated by sera against DEV, Duck hepatitis virus (DHV), Riemerella anatipestifer (RA), Duck E. coli, Muscovy duck parvovirus (MPV), or Duck Influenza viruses (DIV). Only sera against DEV showed the strong positive results. In order to determine the sensitivity of the ICS, anti-DEV serum diluted serially was tested, and the minimum detection limit of 1:128 was obtained. The ICS components, which are provided in a sealed package, require no refrigeration and are stable for 12 months. To evaluate the effect of the ICS, 110 duck serum samples collected from several non-immune duck flocks were simultaneously tested by the ICS test, enzyme-linked immunosorbent assay (ELISA) and neutralization test (NT). The results showed that the sensitivity of the ICS test was almost consistent with ELISA and much higher than NT, has low cost, and is rapid (15 min) and easy to perform with no requirement of specialized equipment, reagent or technicians. Conclusions In this work, we successfully developed a simple and rapid ICS test for detecting DEV serum antibodies for the first time. The ICS test was high specific and sensitive for the rapid detection of anti-DEV antibodies, and has great potential to be used for the serological surveillance of DEV

  4. Radiation enteritis.

    PubMed

    Harb, Ali H; Abou Fadel, Carla; Sharara, Ala I

    2014-01-01

    Radiation enteritis continues to be a major health concern in recipients of radiation therapy. The incidence of radiation enteritis is expected to continue to rise during the coming years paralleling the unprecedented use of radiotherapy in pelvic cancers. Radiation enteritis can present as either an acute or chronic syndrome. The acute form presents within hours to days of radiation exposure and typically resolves within few weeks. The chronic form may present as early as 2 months or as long as 30 years after exposure. Risk factors can be divided into patient and treatment-related factors. Chronic radiation enteritis is characterized by progressive obliterative endarteritis with exaggerated submucosal fibrosis and can manifest by stricturing, formation of fistulae, local abscesses, perforation, and bleeding. In the right clinical context, diagnosis can be confirmed by cross-sectional imaging, flexible or video capsule endoscopy. Present treatment strategies are directed primarily towards symptom relief and management of emerging complications. Recently, however, there has been a shift towards rational drug design based on improved understanding of the molecular basis of disease in an effort to limit the fibrotic process and prevent organ damage.

  5. Detection of somatic coliphages through a bioluminescence assay measuring phage mediated release of adenylate kinase and adenosine 5'-triphosphate.

    PubMed

    Guzmán Luna, Carolina; Costán-Longares, Ana; Lucena, Francisco; Jofre, Joan

    2009-10-01

    The feasibility of detecting somatic coliphages by phage infection of Escherichia coli WG5 and measurement of phage propagation by the lysis mediated release of the bacterial host adenylate kinase (AK) and adenosine 5'-triphosphate (ATP) detected by a bioluminescent signal was evaluated. After 2h of incubation, all cultures infected with reference bacteriophage phiX174 showed a significant increase in the bioluminescent signal, even with number of phages as low as less of 10 plaque forming units (PFU). Naturally occurring somatic coliphages ensured a significant bioluminescent signal after 3h of infection when >10 PFU were inoculated. These results indicate that an easy and reliable method to detect low numbers of coliphages in less than 3h is feasible.

  6. Proteins responsible for lysogenic conversion caused by coliphages N15 and phi80 are highly homologous.

    PubMed Central

    Vostrov, A A; Vostrukhina, O A; Svarchevsky, A N; Rybchin, V N

    1996-01-01

    Lysogenic conversion caused by lambdoid bacteriophage phi80 and that caused by coliphage N15 have similar characteristics, suggesting that similarities in their cor genes and Cor proteins are responsible for this effect. Here we present the nucleotide sequence of the N15 cor gene. The N15 cor gene homolog was found in the phi80 cor region, but in the opposite direction of that of the open reading frame to which the phi80 cor gene had previously been assigned (M. Matsumoto, N. Ichikawa, S. Tanaka, T. Morita, and A. Matsushiro, Jpn. J. Genet. 60:475-483, 1985). PMID:8631731

  7. Toxicity of zinc to fungi, bacteria, and coliphages: influence of chloride ions.

    PubMed Central

    Babich, H; Stotzky, G

    1978-01-01

    A 10 mM concentration of Zn2+ decreased the survival of Escherichia coli; enhanced the survival of Bacillus cereus; did not significantly affect the survival of Pseudomonas aeruginosa, Norcardia corallina, and T1, T7, P1, and phi80 coliphages; completely inhibited mycelial growth of Rhizoctonia solani; and reduced mycelial growth of Fusarium solani, Cunninghamella echinulata, Aspergillus niger, and Trichoderma viride. The toxicity of zinc to the fungi, bacteria, and coliphages was unaffected, lessened, or increased by the addition of high concentrations of NaCl. The increased toxicity of zinc in the presence of high concentrations of NaCl was not a result of a synergistic interaction between Zn2+ and elevated osmotic pressures but of the formation of complex anionic ZnCl species that exerted greater toxicities than did cationic Zn2+. Conversely, the decrease in zinc toxicity with increasing concentrations of NaCl probably reflected the decrease in the levels of Zn2+ due to the formation of Zn-Cl species, which was less inhibitory to these microbes than was Zn2+. A. niger tolerated higher concentrations of zinc in the presence of NaCl at 37 than at 25 degrees C. PMID:736544

  8. Evaluation of a simple and cost effective filter paper-based shipping and storage medium for environmental sampling of F-RNA coliphages.

    PubMed

    Pérez-Méndez, A; Chandler, J C; Bisha, B; Coleman, S M; Zhanqiang, S; Gang, Y; Goodridge, L D

    2013-12-01

    Male specific RNA (F-RNA) coliphages are used as indicators of fecal contamination and for source tracking. However, collecting fecal samples for analysis from remote sites is problematic due to the need for an uninterrupted cold chain to guarantee sample suitability for downstream molecular detection of these coliphages. Here, we investigated the feasibility of using filter paper as a collection and storage vehicle for F-RNA coliphages. Various concentrations (10(1) to 10(4)pfu) of two F-RNA coliphages, MS2 and Qβ, were prepared in lambda buffer or a 10% bovine manure slurry, spotted onto filter paper disks, dried, and maintained at 37 °C for up to 37 days. Nucleic acids were extracted from the spotted filter paper disks at 0, 6, 13, and 37 days post inoculation and analyzed by real time RT-PCR. F-RNA coliphages at concentrations of 10(2)pfu/filter paper unit were readily detected, and only a slight decrease in nucleic acid detection was observed over time. Furthermore, the sensitivity of real time RT-PCR detection of the F-RNA coliphage RNA was similar between the developed filter paper sampling methodology and traditional cold storage. These results indicate that filter paper is a suitable storage and transport medium for F-RNA coliphages when refrigeration is not possible.

  9. Application of microbial risk assessment to the development of standards for enteric pathogens in water used to irrigate fresh produce.

    PubMed

    Stine, Scott W; Song, Inhong; Choi, Christopher Y; Gerba, Charles P

    2005-05-01

    Microbial contamination of the surfaces of cantaloupe, iceberg lettuce, and bell peppers via contact with irrigation water was investigated to aid in the development of irrigation water quality standards for enteric bacteria and viruses. Furrow and subsurface drip irrigation methods were evaluated with the use of nonpathogenic surrogates, coliphage PRD1, and Escherichia coli ATCC 25922. The concentrations of hepatitis A virus (HAV) and Salmonella in irrigation water necessary to achieve a 1:10,000 annual risk of infection, the acceptable level of risk used for drinking water by the U.S. Environmental Protection Agency, were calculated with a quantitative microbial risk assessment approach. These calculations were based on the transfer of the selected nonpathogenic surrogates to fresh produce via irrigation water, as well as previously determined preharvest inactivation rates of pathogenic microorganisms on the surfaces of fresh produce. The risk of infection was found to be variable depending on type of crop, irrigation method, and days between last irrigation event and harvest. The worst-case scenario, in which produce is harvested and consumed the day after the last irrigation event and maximum exposure is assumed, indicated that concentrations of 2.5 CFU/100 ml of Salmonella and 2.5 x 10(-5) most probable number per 100 ml of HAV in irrigation water would result in an annual risk of 1:10,000 when the crop was consumed. If 14 days elapsed before harvest, allowing for die-off of the pathogens, the concentrations were increased to 5.7 x 10(3) Salmonella per 100 ml and 9.9 x 10(-3) HAV per 100 ml.

  10. Enteric Fever.

    PubMed

    Kumar, Praveen; Kumar, Ruchika

    2017-03-01

    Enteric fever is an important public-health problem in India. The clinical presentation of typhoid fever is very variable, ranging from fever with little other morbidities to marked toxemia and associated multisystem complications. Fever is present in majority of patients (>90 %) irrespective of their age group. Mortality is higher in younger children. Blood culture remains gold standard for diagnosis. Widal test has low sensitivity and specificity but may be used in second week to support the diagnosis. Emerging resistance to several antibiotics should be kept in mind when selecting antibiotics or revising the treatment. The key preventive strategies are safe water, safe food, personal hygiene, and appropriate sanitation. Vaccination is an additional effective tool for prevention.

  11. Hand-mouth transfer and potential for exposure to E. coli and F+ coliphage in beach sand, Chicago, Illinois

    USGS Publications Warehouse

    Whitman, R.L.; Przybyla-Kelly, K.; Shively, D.A.; Nevers, M.B.; Byappanahalli, M.N.

    2009-01-01

    Beach sand contains fecal indicator bacteria, often in densities greatly exceeding the adjacent swimming waters. We examined the transferability of Escherichia coli and F+ coliphage (MS2) from beach sand to hands in order to estimate the potential subsequent health risk. Sand with high initial E. coli concentrations was collected from a Chicago beach. Individuals manipulated the sand for 60 seconds, and rinse water was analysed for E. coli and coliphage. E. coli densities transferred were correlated with density in sand rather than surface area of an individual's hand, and the amount of coliphage transferred from seeded sand was different among individuals. In sequential rinsing, percentage reduction was 92% for E. coli and 98% for coliphage. Using dose-response estimates developed for swimming water, it was determined that the number of individuals per thousand that would develop gastrointestinal symptoms would be 11 if all E. coli on the fingertip were ingested or 33 if all E. coli on the hand were ingested. These results suggest that beach sand may be an important medium for microbial exposure; bacteria transfer is related to initial concentration in the sand; and rinsing may be effective in limiting oral exposure to sand-borne microbes of human concern.

  12. A Reverse Transcription-PCR Assay to Distinguish the Four Genogroups of Male-Specific (F+) RNA Coliphages

    EPA Science Inventory

    Goals of reducing fecal contamination in recreational, drinking, shellfishing and other waters and accurately assessing risk from exposure can best be attained if tools to distinguish between sources of pollution are available. The male-specific RNA coliphage (FRNA) genogroups h...

  13. Persistence of somatic and F-specific coliphages, potential indicators of fecal contamination, on spinach foliar tissue

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent outbreaks of foodborne illness have been linked to consumption of fresh leafy greens. E. coli is commonly used as the indicator microorganism for fecal contamination on leafy greens. However, alternative microorganisms, such as coliphages, may deserve consideration as indicators of fecal cont...

  14. Persistence of somatic and F-specific coliphages, potential indicators of fecal contamination, on spinach foliar tissue

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: Recent outbreaks of foodborne illness have been linked to consumption of fresh leafy greens. E. coli is commonly used as the indicator microorganism for fecal contamination on growing leafy greens. However, alternative microorganisms, such as coliphages, may be an alternative indicat...

  15. Accumulation of enteric bacteriophage in fresh water sediments.

    PubMed

    Skraber, Sylvain; Schijven, Jack; Italiaander, Ronald; de Roda Husman, Ana Maria

    2009-09-01

    Our study aimed to assess the accumulation of bacteriophages in sandy and clayey fresh water sediments. All of the 24 natural fresh water sediments were positive for somatic and F-specific phages, though their concentrations in the overlying water were undetectable in 1 and 11 samples, respectively, out of 24, corresponding to 4 and 46% for somatic and F-specific phages, respectively. Based on the sediment-to-water ratios, F-specific phages accumulate over 100 times more than the somatic coliphages in clayey sediments. Inactivation of bacteriophages in clayey and sandy sediments over a 1-month period at 15 degrees C was negligible. Our data suggest that persistence of deposited viruses in fresh water sediments leads to accumulation and the findings call for additional investigations on the fate of entrapped pathogenic viruses.

  16. Sequential concentration of bacteria and viruses from marine waters using a dual membrane system.

    PubMed

    Abdelzaher, A M; Solo-Gabriele, H M; Wright, M E; Palmer, C J

    2008-01-01

    The ability to rapidly and effectively concentrate diverse microbes is an essential component for monitoring water quality at recreational beaches. The purpose of this study was to develop a 0.45 microm pore size dual membrane system, which can sequentially concentrate both viruses and bacteria. The top PVDF membrane was used to filter bacteria by physical straining while the bottom HA membrane retained viruses through adsorption. The recovery of this system was assessed using test organisms: enterococci and somatic coliphage. Volumes of 100 to 400 mL of unspiked and sewage-spiked beach water were filtered through both types of membranes. The PVDF membrane recovered statistically equivalent amounts of enterococci when compared to traditional membranes. All of the coliphage passed through the PVDF membrane, while 22% passed through the HA membrane. Increasing the volume from 100 to 400 mL did not significantly influence recoveries. Up to 35% of coliphage was eluted from the bottom membrane using beef extract solution. Rinsing bottom membranes with 0.5 mmol L(-1) H(2)S0(4) was found to deactivate somatic coliphage. This research demonstrates the potential of using a dual membrane adsorption system for the concentration of both bacteria and viruses from recreational beaches. A proposed bi-layer filtration system can be designed for simultaneous bacteria and virus filtration. Future experiments should focus on measurements utilizing additional bacteria and viruses.

  17. Modeling the infection dynamics of bacteriophages in enteric Escherichia coli: estimating the contribution of transduction to antimicrobial gene spread.

    PubMed

    Volkova, Victoriya V; Lu, Zhao; Besser, Thomas; Gröhn, Yrjö T

    2014-07-01

    Animal-associated bacterial communities are infected by bacteriophages, although the dynamics of these infections are poorly understood. Transduction by bacteriophages may contribute to transfer of antimicrobial resistance genes, but the relative importance of transduction among other gene transfer mechanisms is unknown. We therefore developed a candidate deterministic mathematical model of the infection dynamics of enteric coliphages in commensal Escherichia coli in the large intestine of cattle. We assumed the phages were associated with the intestine and were predominantly temperate. Model simulations demonstrated how, given the bacterial ecology and infection dynamics, most (>90%) commensal enteric E. coli bacteria may become lysogens of enteric coliphages during intestinal transit. Using the model and the most liberal assumptions about transduction efficiency and resistance gene frequency, we approximated the upper numerical limits ("worst-case scenario") of gene transfer through specialized and generalized transduction in E. coli by enteric coliphages when the transduced genetic segment is picked at random. The estimates were consistent with a relatively small contribution of transduction to lateral gene spread; for example, generalized transduction delivered the chromosomal resistance gene to up to 8 E. coli bacteria/hour within the population of 1.47 × 10(8) E. coli bacteria/liter luminal contents. In comparison, the plasmidic blaCMY-2 gene carried by ~2% of enteric E. coli was transferred by conjugation at a rate at least 1.4 × 10(3) times greater than our generalized transduction estimate. The estimated numbers of transductants varied nonlinearly depending on the ecology of bacteria available for phages to infect, that is, on the assumed rates of turnover and replication of enteric E. coli.

  18. Construction of recombinant Newcastle disease virus expressing the S1 protein of Turkey enteric coronavirus for use as a bivalent vaccine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Turkey enteric coronavirus (TCoV) causes a contagious form of enteritis in turkeys, generally recognized in the field by outward signs including diarrhea and decreased weight gain, resulting in severe economic losses for the poultry industry in the US. To date there is no commercial vaccine availab...

  19. Prevalence and Correlates of Genital Infections Among Newly Diagnosed Human Immunodeficiency Virus–Infected Adults Entering Human Immunodeficiency Virus Care in Windhoek, Namibia

    PubMed Central

    Djomand, Gaston; Schlefer, Madeleine; Gutreuter, Steve; Tobias, Sarah; Patel, Roopal; DeLuca, Nickolas; Hood, Julia; Sawadogo, Souleymane; Chen, Cheng; Muadinohamba, Alexinah; Lowrance, David W.; Bock, Naomi

    2016-01-01

    Background Identifying and treating genital infections, including sexually transmitted infections (STI), among newly diagnosed human immunodeficiency virus (HIV)-infected individuals may benefit both public and individual health. We assessed prevalence of genital infections and their correlates among newly diagnosed HIV-infected individuals enrolling in HIV care services in Namibia. Methods Newly diagnosed HIV-infected adults entering HIV care at 2 health facilities in Windhoek, Namibia, were recruited from December 2012 to March 2014. Participants provided behavioral and clinical data including CD4+ T lymphocyte counts. Genital and blood specimens were tested for gonorrhea, Chlamydia, trichomoniasis, Mycoplasma genitalium, syphilis, bacterial vaginosis, and vulvovaginal candidiasis. Results Among 599 adults, 56% were women and 15% reported consistent use of condoms in the past 6 months. The most common infections were bacterial vaginosis (37.2%), trichomoniasis (34.6%) and Chlamydia (14.6%) in women and M. genitalium (11.4%) in men. Correlates for trichomoniasis included being female (adjusted relative risk, [aRR], 7.18; 95% confidence interval [CI], 4.07–12.65), higher education (aRR, 0.58; 95% CI, 0.38–0.89), and lower CD4 cell count (aRR, 1.61; 95% CI, 1.08–2.40). Being female (aRR, 2.39; 95% CI, 1.27–4.50), nonmarried (aRR, 2.30; (95% CI, 1.28–4.14), and having condomless sex (aRR, 2.72; 95% CI, 1.06–7.00) were independently associated with chlamydial infection. Across all infections, female (aRR, 2.31; 95% CI, 1.79–2.98), nonmarried participants (aRR, 1.29; 95% CI, 1.06–1.59), had higher risk to present with any STI, whereas pregnant women (aRR, 1.16, 95% CI 1.03–1.31) were at increased risk of any STI or reproductive tract infection. PMID:27893600

  20. Foodborne viruses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Testing for human pathogenic viruses in foods represents a formidable task requiring the extraction, concentration, and assay of a host of viruses from a wide range of food matrices. The enteric viruses, particularly genogroup I and II (GI and GII) noroviruses and hepatitis A virus, are the princip...

  1. Process Relationships for Evaluating the Role of Light-induced Inactivation of Coliphages at Selected Beaches and Nearby Tributaries of the Great Lakes

    EPA Science Inventory

    Past studies have indicated that sunlight plays an important role in altering densities of coliphages, other indicator microorganisms and pathogens in aquatic environments. Fate and transport modeling of bacteriophage requires mathematical relationships that describe the waveleng...

  2. Structural basis of transcription arrest by coliphage HK022 nun in an Escherichia coli RNA polymerase elongation complex.

    PubMed

    Kang, Jin Young; Olinares, Paul Dominic B; Chen, James; Campbell, Elizabeth A; Mustaev, Arkady; Chait, Brian T; Gottesman, Max E; Darst, Seth A

    2017-03-20

    Coliphage HK022 Nun blocks superinfection by coliphage λ by stalling RNA polymerase (RNAP) translocation specifically on λΔNA.To provide a structural framework to understand how Nun blocks RNAP translocation, we determined structures of Escherichia coli RNAP ternary elongation complexes (TECs) with and without Nun by single-particle cryo-electron microscopy. Nun fits tightly into the TEC by taking advantage of gaps between the RNAP and the nucleic acids. The C-terminal segment of Nun interacts with the RNAP β and β' subunits inside the RNAP active site cleft as well as with nearly every element of the nucleic-acid scaffold, essentially crosslinking the RNAP and the nucleic acids to prevent translocation, a mechanism supported by the effects of Nun amino acid substitutions. The nature of Nun interactions inside the RNAP active site cleft suggests that RNAP clamp opening is required for Nun to establish its interactions, explaining why Nun acts on paused TECs.

  3. Comparative reduction of Giardia cysts, F+ coliphages, sulphite reducing clostridia and fecal coliforms by wastewater treatment processes.

    PubMed

    Nasser, Abidelfatah M; Benisti, Neta-Lee; Ofer, Naomi; Hovers, Sivan; Nitzan, Yeshayahu

    2017-01-28

    Advanced wastewater treatment processes are applied to prevent the environmental dissemination of pathogenic microorganisms. Giardia lamblia causes a severe disease called giardiasis, and is highly prevalent in untreated wastewater worldwide. Monitoring the microbial quality of wastewater effluents is usually based on testing for the levels of indicator microorganisms in the effluents. This study was conducted to compare the suitability of fecal coliforms, F+ coliphages and sulfide reducing clostridia (SRC) as indicators for the reduction of Giardia cysts in two full-scale wastewater treatment plants. The treatment process consists of activated sludge, coagulation, high rate filtration and either chlorine or UV disinfection. The results of the study demonstrated that Giardia cysts are highly prevalent in raw wastewater at an average concentration of 3600 cysts/L. Fecal coliforms, F+ coliphages and SRC were also detected at high concentrations in raw wastewater. Giardia cysts were efficiently removed (3.6 log10) by the treatment train. The greatest reduction was observed for fecal coliforms (9.6 log10) whereas the least reduction was observed for F+ coliphages (2.1 log10) following chlorine disinfection. Similar reduction was observed for SRC by filtration and disinfection by either UV (3.6 log10) or chlorine (3.3 log10). Since F+ coliphage and SRC were found to be more resistant than fecal coliforms for the tertiary treatment processes, they may prove to be more suitable as indicators for Giardia. The results of this study demonstrated that advanced wastewater treatment may prove efficient for the removal of Giardia cysts and may prevent its transmission when treated effluents are applied for crop irrigation or streams restoration.

  4. Postharvest Reduction of Coliphage MS2 from Romaine Lettuce during Simulated Commercial Processing with and without a Chlorine-Based Sanitizer.

    PubMed

    Wengert, Samantha L; Aw, Tiong Gim; Ryser, Elliot T; Rose, Joan B

    2017-02-01

    Viral foodborne outbreaks are a serious threat to public health, and fresh produce is becoming increasingly recognized as a transmission vehicle. To limit foodborne disease, ready-to-eat leafy greens are typically washed with a chlorine-based sanitizer during commercial production. This study assessed the efficacy of a chlorine-based sanitizer against coliphage MS2, as a potential surrogate for foodborne viruses, on fresh-cut romaine lettuce during simulated commercial production using a small-scale processing line. Before processing, romaine lettuce was inoculated to contain approximately 10(5) and 10(6) PFU/g of MS2 for experiments with and without sanitizer, respectively. Lettuce samples were collected following each stage of processing, which included mechanical shredding, 2 min of flume washing (with or without 25 ppm of free chlorine), shaker table dewatering, and centrifugal drying. In addition, the spent centrifuge water and flume wash water were collected, with the flume water concentrated using hollow-fiber ultrafiltration. MS2 was recovered from lettuce in Tris-glycine buffer and quantified as PFUs in a double-agar overlay assay. The greatest reduction in MS2 occurred between shredding and flume washing, with levels remaining relatively stable following flume washing with or without 25 ppm of free chlorine. Average total reductions of 0.8 and 1.0 log PFU/g were seen after processing with and without the sanitizer, respectively, with no statistical difference observed between the two treatments (P > 0.05). The average MS2 level in the spent centrifugation water started at 4.0 log PFU/ml for experiments with sanitizer and the average MS2 reduction in the flume wash water was 4 log (PFU) for experiments with sanitizer, demonstrating that removals could be achieved in the water itself. These findings suggest that the currently recommended commercial production practices are unable to effectively decrease viruses once they have attached to leafy greens during

  5. Canine viral enteritis. Recent developments.

    PubMed

    Pollock, R V; Carmichael, L

    1979-05-01

    Two apparently novel viral gastroenteritides of dogs were recognized in 1978: one caused by a parvo-like virus (CPV) and one by a corona-like virus (CCV). A rotavirus has also been tentatively associated with neonatal pup enteritis. Canine viral enteritis is characterized by a sudden onset of vomiting and diarrhea, rapid spread and high morbidity. Treatment is only supportive but must be initiated promptly. Infected animals should be isolated immediately; the extremely contagious nature of these diseases makes them difficult to contain. Feces from infected dogs appear to be the primary means of transmission. Sodium hypochlorite solutions (eg, Clorox) are recommended for disinfection. The development of effective vaccines is an immediate and pressing problem.

  6. Propidium Monoazide Integrated with qPCR Enables the Detection and Enumeration of Infectious Enteric RNA and DNA Viruses in Clam and Fermented Sausages

    PubMed Central

    Quijada, Narciso M.; Fongaro, Gislaine; Barardi, Célia R. M.; Hernández, Marta; Rodríguez-Lázaro, David

    2016-01-01

    The increase of foodborne viral outbreaks highlights the need for a rapid and sensitive method for the prediction of viral infectivity in food samples. This study assesses the use of propidium monoazide (PMA) coupled with real-time PCR methods (RT-qPCR or qPCR for RNA or DNA viruses, respectively) in the determination of viral infectivity in complex animal-related food matrices. Clam and Spanish fermented sausage (“chorizo”) samples were spiked with infectious and heat-inactivated human adenovirus-2 (HAdV-2) and mengovirus (vMC0). PMA-qPCR/RT-qPCR discriminated infective virus particles, with significant reductions (>2.7 log10 or 99.7%). Additionally, infectious HAdV-2 and vMC0 were quantified by plaque assay (in plaque forming units, PFU), and compared with those in virus genomes copies (GCs) quantified by PMA-qPCR/RT-qPCR. A consistent correlation (R2 > 0.92) was showed between PFU and GCs along serial 10-fold dilutions in both DNA and RNA virus and in both food matrices. This study shows the use of PMA coupled to qPCR/RT-qPCR as a promising alternative for prediction of viral infectivity in food samples in comparison to more expensive and time-consuming methods and for those viruses that are not able to grow under available cell culture techniques. PMID:28018329

  7. The blood-brain barrier in the cerebrum is the initial site for the Japanese encephalitis virus entering the central nervous system.

    PubMed

    Liu, Tsan-Hsiun; Liang, Li-Ching; Wang, Chien-Chih; Liu, Huei-Chung; Chen, Wei-June

    2008-11-01

    Japanese encephalitis (JE) virus is a member of the encephalitic flaviviruses and frequently causes neurological sequelae in a proportion of patients who survive the acute phase of the infection. In the present study, we molecularly identified viral infection in the brain of mice with rigidity of hindlimbs and/or abnormal gait, in which JE virus particles appeared within membrane-bound vacuoles of neurons throughout the central nervous system. Deformation of tight junctions (TJs) shown as dissociation of endothelial cells in capillaries, implying that the integrity of the blood-brain barrier (BBB) has been compromised by JE virus infection. BBB permeability evidently increased in the cerebrum, but not in the cerebellum, of JE virus-infected mice intravenously injected with the tracer of Evans blue dye. This suggests that the permeability of the BBB differentially changed in response to viral infection, leading to the entry of JE virions and/or putatively infected leukocytes from the periphery to the cerebrum as the initial site of infection in the central nervous system (CNS). Theoretically, the virus spread to the cerebellum soon after the cerebrum became infected.

  8. Propidium Monoazide Integrated with qPCR Enables the Detection and Enumeration of Infectious Enteric RNA and DNA Viruses in Clam and Fermented Sausages.

    PubMed

    Quijada, Narciso M; Fongaro, Gislaine; Barardi, Célia R M; Hernández, Marta; Rodríguez-Lázaro, David

    2016-01-01

    The increase of foodborne viral outbreaks highlights the need for a rapid and sensitive method for the prediction of viral infectivity in food samples. This study assesses the use of propidium monoazide (PMA) coupled with real-time PCR methods (RT-qPCR or qPCR for RNA or DNA viruses, respectively) in the determination of viral infectivity in complex animal-related food matrices. Clam and Spanish fermented sausage ("chorizo") samples were spiked with infectious and heat-inactivated human adenovirus-2 (HAdV-2) and mengovirus (vMC0). PMA-qPCR/RT-qPCR discriminated infective virus particles, with significant reductions (>2.7 log10 or 99.7%). Additionally, infectious HAdV-2 and vMC0 were quantified by plaque assay (in plaque forming units, PFU), and compared with those in virus genomes copies (GCs) quantified by PMA-qPCR/RT-qPCR. A consistent correlation (R(2) > 0.92) was showed between PFU and GCs along serial 10-fold dilutions in both DNA and RNA virus and in both food matrices. This study shows the use of PMA coupled to qPCR/RT-qPCR as a promising alternative for prediction of viral infectivity in food samples in comparison to more expensive and time-consuming methods and for those viruses that are not able to grow under available cell culture techniques.

  9. Biomarkers in canine parvovirus enteritis.

    PubMed

    Schoeman, J P; Goddard, A; Leisewitz, A L

    2013-07-01

    Canine parvovirus (CPV) enteritis has, since its emergence in 1978, remained a common and important cause of morbidity and mortality in young dogs. The continued incidence of parvoviral enteritis is partly due to the virus' capability to evolve into more virulent and resistant variants with significant local gastrointestinal and systemic inflammatory sequelae. This paper reviews current knowledge on historical-, signalment-, and clinical factors as well as several haematological-, biochemical- and endocrine parameters that can be used as diagnostic and prognostic biomarkers in CPV enteritis. These factors include season of presentation, purebred nature, bodyweight, vomiting, leukopaenia, lymphopaenia, thrombocytopaenia, hypercoagulability, hypercortisolaemia, hypothyroxinaemia, hypoalbuminaemia, elevated C-reactive protein and tumour necrosis factor, hypocholesterolaemia and hypocitrullinaemia. Factors contributing to the manifestations of CPV infection are multiple with elements of host, pathogen, secondary infections, underlying stressors and environment affecting severity and outcome. The availability of several prognosticators has made identification of patients at high risk of death and their subsequent targeted management more rewarding.

  10. Occurrence of viruses and protozoa in drinking water sources of Japan and their relationship to indicator microorganisms.

    PubMed

    Haramoto, Eiji; Kitajima, Masaaki; Kishida, Naohiro; Katayama, Hiroyuki; Asami, Mari; Akiba, Michihiro

    2012-09-01

    A nationwide survey of viruses, protozoa, and indicator microorganisms in drinking water sources of Japan was conducted. Among 64 surface water samples collected from 16 drinking water treatment plants, 51 (80 %) samples were positive for at least one of the 11 pathogen types tested, including noroviruses of genogroups I (positive rate, 13 %) and II (2 %), human sapoviruses (5 %), human adenoviruses of serotypes 40 and 41 (39 %), Cryptosporidium oocysts (41 %), and Giardia cysts (36 %). Total coliforms, Escherichia coli, and F-specific coliphages were detected in 63 (98 %), 33 (52 %), and 17 (27 %) samples, respectively, and E. coli was judged to be the most suitable indicator of pathogen contamination of drinking water sources. Genogroup-specific real-time PCR for F-specific coliphages revealed the presence of F-specific RNA coliphages of animal genogroup I and human genogroups II and III in 13 (41 %), 12 (39 %), and 1 (3 %), respectively, of 31 plaques isolated.

  11. Inactivation of MS2 coliphage by ferrous ion and zero-valent iron nanoparticles.

    PubMed

    Kim, Jee Yeon; Lee, Changha; Love, David C; Sedlak, David L; Yoon, Jeyong; Nelson, Kara L

    2011-08-15

    This study demonstrates the inactivation of MS2 coliphage (MS2) by nano particulate zerovalent iron (nZVI) and ferrous ion (Fe[II]) in aqueous solution. For nZVI, the inactivation efficiency of MS2 under air-saturated conditions was greater than that observed under deaerated conditions, indicating that reactions associated with the oxidation of nZVI were mainly responsible for the MS2 inactivation. Under air-saturated conditions, the inactivation efficiency increased with decreasing pH for both nZVI and Fe(II), associated with the pH-dependent stability of Fe(II). Although the Fe(II) released from nZVI appeared to contribute significantly to the virucidal activity of nZVI, several findings suggest that the nZVI surfaces interacted directly with the MS2 phages, leading to their inactivation. First, the addition of 1,10-phenanthroline (a strong Fe(II)-chelating agent) failed to completely block the inactivation of MS2 by nZVI. Second, under deaerated conditions, a linear dose-log inactivation curve was still observed for nZVI. Finally, ELISA analysis indicated that nZVI caused more capsid damage than Fe(II).

  12. Involvement of the Host Initiator Function Dnaa in the Replication of Coliphage λ

    PubMed Central

    Wegrzyn, G.; Wegrzyn, A.; Konieczny, I.; Bielawski, K.; Konopa, G.; Obuchowski, M.; Helinski, D. R.; Taylor, K.

    1995-01-01

    We demonstrate that the initiation of coliphage λ DNA replication is dependent on the host initiator function dnaA, provided that the lambdoid prophage Rac is absent. Presence of Rac compensated the absence of dnaA function, causing initiation of replication. In dnaAts rac(+) cells at 43°, most of parental phage DNA molecules, after one round of theta replication, switched to a replication with features of the sigma mode and produced progeny at high yield. Initiation of replication of the λPts1 mutant at 43° was blocked by dnaA function; however, under dnaA(-)rac(+) conditions all parental phage DNA molecules, after one round of theta replication, switched to the sigma mode and produced progeny at high yield. Taking into account our recent finding that transcriptional activation of oriλ seems to be dnaA-regulated (to be published elsewhere), we suggest that the DnaA-λPts1 incompatibility occurs at the insertion of the oriλ-bound λO-λP-DnaB preprimosome between the complementary λ DNA strands. The role of Rac and the mechanism of the switch from theta to sigma mode of λ phage DNA replication are discussed. PMID:7789753

  13. Persistence of F-Specific RNA Coliphages in Surface Waters from a Produce Production Region along the Central Coast of California

    PubMed Central

    Ravva, Subbarao V.; Sarreal, Chester Z.

    2016-01-01

    F+ RNA coliphages (FRNA) are used to source-track fecal contamination and as surrogates for enteric pathogen persistence in the environment. However, the environmental persistence of FRNA is not clearly understood and necessitates the evaluation of the survival of prototype and environmental isolates of FRNA representing all four genogroups in surface waters from the central coast of California. Water temperature played a significant role in persistence–all prototype and environmental strains survived significantly longer at 10°C compared to 25°C. Similarly, the availability of host bacterium was found to be critical in FRNA survival. In the absence of E. coli Famp, all prototypes of FRNA disappeared rapidly with a D-value (days for one log reduction) of <1.2 d from water samples incubated at 25°C; the longest surviving prototype was SP. However, in the presence of the host, the order of persistence at 25°C was QB>MS2>SP>GA and at 10°C it was QB = MS2>GA>SP. Significant differences in survival were observed between prototypes and environmental isolates of FRNA. While most environmental isolates disappeared rapidly at 25°C and in the absence of the host, members of genogroups GIII and GI persisted longer with the host compared to members of GII and GIV. Consequentially, FRNA based source tracking methods can be used to detect phages from recent fecal contamination along with those that persist longer in the environment as a result of cooler temperatures and increased host presence. PMID:26784030

  14. Investigating turkey enteric picornavirus and its association with enteric disease in poults

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Our previous research into the dynamic viral community in the turkey gastrointestinal tract revealed a number of novel enteric viruses. Of particular note in this previous metagenomic investigation was the observation of a number of novel avian enteric picornaviruses, in addition to numerous other ...

  15. Removal of Viruses in Drinking Water- Ultrafiltration Module with a Cut Fiber, Dow Chemical Company - Dow Water & Process Solutions, SFD-2880 - Ultrafiltration Module

    EPA Science Inventory

    The purpose of this verification was a cut fiber challenge study for the Dow Chemical Company SFD-2880 UF membrane module. MS2 coliphage virus was the surrogate challenge organism. The challenge tests followed the requirements of the Department of Health Victoria (Australia) Dr...

  16. Phylogenetic Diversity of Marine Cyanophage Isolates and Natural Virus Communities as Revealed by Sequences of Viral Capsid Assembly Protein Gene g20†

    PubMed Central

    Zhong, Yan; Chen, Feng; Wilhelm, Steven W.; Poorvin, Leo; Hodson, Robert E.

    2002-01-01

    In order to characterize the genetic diversity and phylogenetic affiliations of marine cyanophage isolates and natural cyanophage assemblages, oligonucleotide primers CPS1 and CPS8 were designed to specifically amplify ca. 592-bp fragments of the gene for viral capsid assembly protein g20. Phylogenetic analysis of isolated cyanophages revealed that the marine cyanophages were highly diverse yet more closely related to each other than to enteric coliphage T4. Genetically related marine cyanophage isolates were widely distributed without significant geographic segregation (i.e., no correlation between genetic variation and geographic distance). Cloning and sequencing analysis of six natural virus concentrates from estuarine and oligotrophic offshore environments revealed nine phylogenetic groups in a total of 114 different g20 homologs, with up to six clusters and 29 genotypes encountered in a single sample. The composition and structure of natural cyanophage communities in the estuary and open-ocean samples were different from each other, with unique phylogenetic clusters found for each environment. Changes in clonal diversity were also observed from the surface waters to the deep chlorophyll maximum layer in the open ocean. Only three clusters contained known cyanophage isolates, while the identities of the other six clusters remain unknown. Whether or not these unidentified groups are composed of bacteriophages that infect different Synechococcus groups or other closely related cyanobacteria remains to be determined. The high genetic diversity of marine cyanophage assemblages revealed by the g20 sequences suggests that marine viruses can potentially play important roles in regulating microbial genetic diversity. PMID:11916671

  17. Structural basis of transcription arrest by coliphage HK022 Nun in an Escherichia coli RNA polymerase elongation complex

    PubMed Central

    Kang, Jin Young; Olinares, Paul Dominic B; Chen, James; Campbell, Elizabeth A; Mustaev, Arkady; Chait, Brian T; Gottesman, Max E; Darst, Seth A

    2017-01-01

    Coliphage HK022 Nun blocks superinfection by coliphage λ by stalling RNA polymerase (RNAP) translocation specifically on λ DNA. To provide a structural framework to understand how Nun blocks RNAP translocation, we determined structures of Escherichia coli RNAP ternary elongation complexes (TECs) with and without Nun by single-particle cryo-electron microscopy. Nun fits tightly into the TEC by taking advantage of gaps between the RNAP and the nucleic acids. The C-terminal segment of Nun interacts with the RNAP β and β’ subunits inside the RNAP active site cleft as well as with nearly every element of the nucleic acid scaffold, essentially crosslinking the RNAP and the nucleic acids to prevent translocation, a mechanism supported by the effects of Nun amino acid substitutions. The nature of Nun interactions inside the RNAP active site cleft suggests that RNAP clamp opening is required for Nun to establish its interactions, explaining why Nun acts on paused TECs. DOI: http://dx.doi.org/10.7554/eLife.25478.001 PMID:28318486

  18. Disinfection efficiency of peracetic acid (PAA): inactivation of coliphages and bacterial indicators in a municipal wastewater plant.

    PubMed

    Zanetti, F; De Luca, G; Sacchetti, R; Stampi, S

    2007-11-01

    The aim of the study was to assess the efficiency of low doses of peracetic acid against viral and bacterial indicators in wastewater and to evaluate if the treatment allows regulatory requirements to be satisfied. A total of 31 samplings were carried out, each involving the collection of secondary effluent and of effluent disinfected with 1.2 or 1.5 mg l(-1) of peracetic acid (contact time 20 minutes). In each sample were measured: somatic coliphages, F-specific RNA bacteriophages, Escherichia coli, total and faecal coliforms, enterococci. Peracetic acid disinfection showed significant differences between the reductions of the microorganisms tested: E. coli showed the highest reduction (1.78 and 2.43 Log respectively with 1.2 and 1.5 mg l(-1) of peracetic acid) and phages the lowest (ranging between 0.52 and 0.60 Log). Only a concentration of 1.5 mg l(-1) of peracetic acid would enable the effluent to be discharged into surface waters in compliance with Italian regulations. The variability of microbial resistance against the peracetic acid disinfection treatment, underlines the importance of assessing disinfection efficiency by using more than one indicator microorganism. The detection of E. coli could be usefully accompanied by tests for more resistant microorganisms such as enterococci or coliphages. In conclusion, peracetic acid can be used for the disinfection of effluents even at low doses, with the advantage of reducing costs and preventing the formation of significant amounts of genotoxic by-products.

  19. Gastric attaching and effacing Escherichia coli lesions in a puppy with naturally occurring enteric colibacillosis and concurrent canine distemper virus infection.

    PubMed

    Wada, Y; Kondo, H; Nakaoka, Y; Kubo, M

    1996-11-01

    A puppy suffering from chronic diarrhea was humanely killed at 90 days of age. Numerous Gram-negative bacilli were found adhering to the surface of as well as within epithelial cells from the stomach to the colon. Canine distemper virus inclusions were in the epithelial cytoplasm of the esophageal, gastric, and intestinal mucosa. Typical attaching and effacing ultrastructural lesions were in the stomach, and some bacilli were in the cytoplasm of the epithelial cells. Escherichia coli, isolated from the contents of the small intestine, belonged to serotype 0118: NM and were negative for plasmid-encoded EPEC adherence factor (EAF) and positive for the E. coli attaching effacing (eae) gene. Immunohistologically, bacilli attached to the epithelium from the stomach to the colon were positive for antisera against E. coli 0118. E. coli 0118: NM inoculated into human tissue culture cells (HEp-2 cells) were attached to the surface of the cells and within the cytoplasm. This is the first report of attaching and effacing E. coli (AEEC) infection in the stomach of the dog.

  20. Immunoprotection against influenza H5N1 virus by oral administration of enteric-coated recombinant Lactococcus lactis mini-capsules.

    PubMed

    Lei, Han; Xu, Yuhong; Chen, Jian; Wei, Xiaohui; Lam, Dominic Man-Kit

    2010-11-25

    Edible vaccines that can be made widely available and easily administered could bring great benefit to the worldwide battle against pandemic viral infections. They can be used not only for the vaccination of humans and domesticated animals, but also for wild herds and live stock which are otherwise difficult to vaccinate. In this study, we report the development of an edible mini-capsule form of live, non-persisting, recombinant Lactococcus lactis (L. lactis) vaccine against the highly virulent influenza H5N1 strain. Recombinant L. lactis-based H5N1 HA antigen expression constructs were made and shown to be able to induce higher levels of HA-specific serum IgG and fecal IgA antibody production after oral administration. The vectors were then formulated into a mini-capsule dosage form and fed to mouse. Four doses of oral administration rendered complete protection of the mouse against lethal challenges of H5N1 virus.

  1. Gene Mapping and Phylogenetic Analysis of the Complete Genome from 30 Single-Stranded RNA Male-Specific Coliphages (Family Leviviridae)

    EPA Science Inventory

    Male-specific ssRNA (FRNA) coliphages belong to the family Leviviridae. Two genera subdivided into four genogroups (Levivirus, genogroups I and II and Allolevivirus, genogroups III and IV) comprise the lower taxonomic levels. Strains isolated from several continents have been p...

  2. Design and Assessment of a Real Time Reverse Transcription-PCR Method to Genotype Single-Stranded RNA Male-Specific Coliphages (Family Leviviridae).

    EPA Science Inventory

    A real-time, reverse transcription-PCR (RT-qPCR) assay was developed to differentiate the four genogroups of male-specific ssRNA coliphages (FRNA) (family Leviviridae). As FRNA display a trend of source-specificity (human sewage or animal waste) at the genogroup level, this assa...

  3. Proposed modifications of Environmental Protection Agency Method 1601 for detection of coliphages in drinking water, with same-day fluorescence-based detection and evaluation by the performance-based measurement system and alternative test protocol validation approaches.

    PubMed

    Salter, Robert S; Durbin, Gregory W; Conklin, Ernestine; Rosen, Jeff; Clancy, Jennifer

    2010-12-01

    Coliphages are microbial indicators specified in the Ground Water Rule that can be used to monitor for potential fecal contamination of drinking water. The Total Coliform Rule specifies coliform and Escherichia coli indicators for municipal water quality testing; thus, coliphage indicator use is less common and advances in detection methodology are less frequent. Coliphages are viral structures and, compared to bacterial indicators, are more resistant to disinfection and diffuse further distances from pollution sources. Therefore, coliphage presence may serve as a better predictor of groundwater quality. This study describes Fast Phage, a 16- to 24-h presence/absence modification of U.S. Environmental Protection Agency (EPA) Method 1601 for detection of coliphages in 100 ml water. The objective of the study is to demonstrate that the somatic and male-specific coliphage modifications provide results equivalent to those of Method 1601. Five laboratories compared the modifications, featuring same-day fluorescence-based prediction, to Method 1601 by using the performance-based measurement system (PBMS) criterion. This requires a minimum 50% positive response in 10 replicates of 100-ml water samples at coliphage contamination levels of 1.3 to 1.5 PFU/100 ml. The laboratories showed that Fast Phage meets PBMS criteria with 83.5 to 92.1% correlation of the same-day rapid fluorescence-based prediction with the next-day result. Somatic coliphage PBMS data are compared to manufacturer development data that followed the EPA alternative test protocol (ATP) validation approach. Statistical analysis of the data sets indicates that PBMS utilizes fewer samples than does the ATP approach but with similar conclusions. Results support testing the coliphage modifications by using an EPA-approved national PBMS approach with collaboratively shared samples.

  4. Prevalence of positive antibody test results for canine parvovirus (CPV) and canine distemper virus (CDV) and response to modified live vaccination against CPV and CDV in dogs entering animal shelters.

    PubMed

    Litster, Annette; Nichols, Jamieson; Volpe, Allison

    2012-05-25

    Canine parvovirus (CPV) and canine distemper virus (CDV) infections are relatively common in animal shelters and are important population management issues since the immune status of incoming dogs is usually unknown. This study aimed to determine the prevalence of positive antibody test results for CPV and CDV in incoming dogs aged ≥ 4 months and to measure antibody response over 2 weeks following vaccination with a modified live vaccine (MLV). Dogs aged 4-24 months entering an adoption-guarantee shelter (Shelter 1, n=51) and aged ≥ 4 months entering a limited admission shelter (Shelter 2; n=51) were enrolled. Dogs from Shelter 1 had been vaccinated with MLV at a municipal shelter 5 days before enrollment, whereas dogs from Shelter 2 had no known history of vaccination at enrollment. Sera were obtained on day 1, immediately prior to CPV/CDV MLV, and tested using an in-clinic ELISA kit to detect CPV/CDV antibodies. Dogs negative for CPV and/or CDV were retested at day 6-8 and those dogs still negative at day 6-8 were retested at day 13-15. Prior to CPV/CDV MLV on day 1, more dogs tested positive for CPV (Shelter 1 - 68.6%; Shelter 2 - 84.3%) than for CDV (Shelter 1 - 37.3%; Shelter 2 - 41.2%). On day 1, prior to MLV, all spayed/neutered animals tested CPV antibody-positive (n=17/102) and CPV antibody-positive dogs were older than serologically negative dogs (Shelter 1, P=0.0029; Shelter 2, P=0.0042). By day 13-15, almost all dogs were CPV antibody-positive (Shelter 1 - 97.9%; Shelter 2 - 100.0%) and CDV antibody-positive (Shelter 1 - 93.8%; Shelter 2 - 97.8%). MLV induces protective antibody titers against CPV/CDV in almost all dogs after 13-15 days.

  5. Diagnostics and intervention strategies for the control and prevention of viral enteric diseases in poultry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent enteric virus surveys have revealed the presence of multiple viruses in turkeys and chickens in the United States, often in samples obtained from the same flock or bird. Complicating the picture considerably is the fact that many of the viruses implicated in enteric disease syndromes are rout...

  6. Pediatric enteral nutrition.

    PubMed

    Axelrod, David; Kazmerski, Kimberly; Iyer, Kishore

    2006-01-01

    Common to all pediatric patients receiving enteral nutrition is the inability to consume calories orally. This is often secondary to issues of inadequate weight gain, inadequate growth, prolonged feeding times, weight loss, a decrease in weight/age or weight/height ratios, or a persistent triceps skinfold thickness <5% for age. Enteral nutrition requires enteral access. In the neonatal period the nasoenteric route is usually used. In pediatric patients requiring long-term enteral access, surgically, endoscopically, or radiologically placed percutaneous feeding tubes are common. Jejunal feeding tubes are used in pediatric patients with gastric feeding intolerance or persistent gastroesophageal reflux. Low-profile enteral access devices are preferred by most pediatric patients because of their cosmetic appearance. For most children, a standard pediatric polypeptide enteral formula is well tolerated. There are specialized pediatric enteral formulas available for patients with decreased intestinal length, altered intestinal absorptive capacity, or altered pancreatic function. Weaning patients from tube feeding to oral nutrition is the ultimate nutrition goal. A multidisciplinary approach to patients with short bowel syndrome will maximize the use of enteral nutrition while preserving parenteral nutrition for patients with true enteral nutrition therapy failure.

  7. Enhanced recovery of airborne T3 coliphage and Pasteurella pestis bacteriophage by means of a presampling humidification technique.

    PubMed

    Hatch, M T; Warren, J C

    1969-05-01

    This paper reports a series of experiments in which two methods of collecting airborne bacteriophage particles were compared. A standard aerosol sampler, the AGI-30, was evaluated for its competence in measuring the content of bacteriophage aerosols. It was used alone or with a prewetting or humidification device (humidifier bulb) to recover T(3) coliphage and Pasteurella pestis bacteriophage particles from aerosols maintained at 21 C and varied relative humidity. Collection of bacteriophage particles via the humidifier bulb altered both the initial recovery level and the apparent biological decay. Sampling airborne bacteriophage particles by the AGI-30 alone yielded data that apparently underestimated the maximal number of potentially viable particles within the aerosol, sometimes by as much as 3 logs.

  8. Viruses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Lytic bacteriophages, viruses which infect and lyse bacterial cells, can provide a natural method to reduce bacterial pathogens on produce commodities. The use of multi-phage cocktails is most likely to be effective against bacterial pathogens on produce commodities, and minimize the development of...

  9. Sequence analysis of parvoviruses associated with enteric disease of poultry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Poult Enteritis Mortality Syndrome (PEMS) and Runting-Stunting Syndrome (RSS) are significant viral enteric diseases of poultry. The etiology of these diseases is not completely understood. Here, we report the application of a molecular screening method that was designed to detect novel viruses from...

  10. [Home enteral nutrition].

    PubMed

    Virgili, N; Vilarasau, M C

    1999-04-01

    Enteral nutrition in the home is applied to stabilized patients who do not require hospitalization or to chronically ill patients who can stay in their homes. However, ensuring the correct administration of this treatment requires a coordinated, expert multidisciplinary team. This article reviews the conditions for use of enteral nutrition in the home, the means of access, the nutritional formulas, the administrative technique, and the complications enteral nutrition in the home may present. Furthermore, the composition and characteristics of the multidisciplinary team which will be in charge of carrying out this treatment is discussed.

  11. [Viral hepatitis of enteric origin].

    PubMed

    Debord, T; Buisson, Y

    1998-01-01

    Hepatitis viruses of oral-fecal origin are responsible for a high morbidity and mortality throughout the world, even if they never result in chronic hepatitis. Two viruses, the virus of hepatitis A (VHA) and of hepatitis E (VHE) are at present the cause of severe viral hepatitis of enteric origin. Water is the principle vector in the spread of these viruses. However, the epidemiological aspects vary according to the pathogenic agent. VHA is excreted in a highly concentrated form in the feces for a relatively short period of time. Since it resists in an exterior environment, the virus remains infectious for a long time. VHE is excreted for a short period of time and in low concentrations. The viral particles are fragile in vitro and their variability in the environment is little known. The possible reservoir role of certain animals has been envisaged. Epidemics arise especially in countries suffering from poor hygiene and massive water pollution. Hepatitis A should no longer be considered a benign disease of childhood. The progress made in hygiene and economic development in industrialized countries have made contacts with this virus scarce, rendering the populations more receptive to it and epidemics more widespread. When the sickness occurs later in life, infection is more often symptomatic and can be serious, resulting sometimes long-term indisposition. Hepatitis E has a vast distribution throughout the world and manifests itself either in epidemic or endemic-sporadic form in many poor countries. In developed countries, it comes about mostly as a result of imported pathology, even if there exists a "substratum" of infection in these areas. The main clinical aspects, such as we were able to study them in 39 cases of military men from Tchad, Guyana and Somalia, are comparable to those of hepatitis A. The reasons for the particular gravity of symptoms in pregnant women are unknown. These affections have no specific treatment. In the field of prevention, vaccination

  12. Strategies for design and application of enteric viral vaccines.

    PubMed

    Chattha, Kuldeep S; Roth, James A; Saif, Linda J

    2015-01-01

    Enteric viral infections in domestic animals cause significant economic losses. The recent emergence of virulent enteric coronaviruses [porcine epidemic diarrhea virus (PEDV)] in North America and Asia, for which no vaccines are available, remains a challenge for the global swine industry. Vaccination strategies against rotavirus and coronavirus (transmissible gastroenteritis virus) infections are reviewed. These vaccination principles are applicable against emerging enteric infections such as PEDV. Maternal vaccines to induce lactogenic immunity, and their transmission to suckling neonates via colostrum and milk, are critical for early passive protection. Subsequently, in weaned animals, oral vaccines incorporating novel mucosal adjuvants (e.g., vitamin A, probiotics) may provide active protection when maternal immunity wanes. Understanding intestinal and systemic immune responses to experimental rotavirus and transmissible gastroenteritis virus vaccines and infection in pigs provides a basis and model for the development of safe and effective vaccines for young animals and children against established and emerging enteric infections.

  13. Enteric viral infections in lambs or kids.

    PubMed

    Martella, V; Decaro, N; Buonavoglia, C

    2015-12-14

    Diarrhoea in lambs and kids is often a complex, multi-factorial syndrome. Common infectious causes of diarrhoea in lambs and kids during the first month of life are of bacterial or parasite nature. However, despite appreciable improvements in management practices and prevention and treatment strategies over the last decades, diarrhoea is still a common and costly syndrome affecting newborn small ruminants. Recent advances in the diagnostics and metagenomic investigations of the enteric environment have allowed discovering a number of novel viruses, although their pathobiological properties remain largely unknown. Assessing more in depth the impact of these viruses on the health and productions of these livestock animals is necessary and requires the development of accurate diagnostic tools and updating of the diagnostic algorithms of enteric pathological conditions.

  14. Simultaneous comparison of murine norovirus, feline calicivirus, coliphage MS2, and GII.4 norovirus to evaluate the efficacy of sodium hypochlorite against human norovirus on a fecally soiled stainless steel surface.

    PubMed

    Park, Geun Woo; Sobsey, Mark D

    2011-09-01

    Free chlorine as hypochlorite is recommended to decontaminate fecally contaminated surfaces to control human norovirus (NoV). We evaluated the efficacy of sodium hypochlorite to decontaminate GII.4 NoV and three surrogates of human NoVs, feline calicivirus (FCV), murine norovirus (MNV), and coliphage MS2, on a fecally soiled stainless steel surface. Reduction of infectivity of FCV, MNV, and MS2 was measured by plaque assay and the decline of genomic copy numbers of GII.4 NoV by reverse transcriptase-polymerase chain reaction. Sodium hypochlorite solution at 5000 ppm could inactivate FCV by 3 log(10) plaque forming units after approximately 1.9 minutes of contact time, but required longer exposure times of 3.2 and 4.5 minutes to reduce MNV and MS2 by 3 log(10), respectively. However, detection of viral RNA by reverse transcriptase-polymerase chain reaction assay may not be reliable to estimate the effectiveness of sodium hypochlorite against human NoV. Of three NoV surrogates, FCV is not the most resistant of the virus tested for inactivation by hypochlorite and thus is not the worst-case model for estimating NoV inactivation. Although the use of 5000 ppm of hypochlorite for fecally soiled surfaces is effective, it may require longer exposure times of ≥3 minutes to control NoVs. Surface precleaning before hypochlorite disinfection is recommended to initially reduce the fecal organic load for better virus inactivation and should be a part of the environmental hygiene response measures during an NoV outbreak or where NoV fecal contamination of environmental surfaces is likely or suspected to be present.

  15. Efficiency of peracetic acid in inactivating bacteria, viruses, and spores in water determined with ATP bioluminescence, quantitative PCR, and culture-based methods.

    PubMed

    Park, Eunyoung; Lee, Cheonghoon; Bisesi, Michael; Lee, Jiyoung

    2014-03-01

    The disinfection efficiency of peracetic acid (PAA) was investigated on three microbial types using three different methods (filtration-based ATP (adenosine-triphosphate) bioluminescence, quantitative polymerase chain reaction (qPCR), culture-based method). Fecal indicator bacteria (Enterococcus faecium), virus indicator (male-specific (F(+)) coliphages (coliphages)), and protozoa disinfection surrogate (Bacillus subtilis spores (spores)) were tested. The mode of action for spore disinfection was visualized using scanning electron microscopy. The results indicated that PAA concentrations of 5 ppm (contact time: 5 min), 50 ppm (10 min), and 3,000 ppm (5 min) were needed to achieve 3-log reduction of E. faecium, coliphages, and spores, respectively. Scanning electron microscopy observation showed that PAA targets the external layers of spores. The lower reduction rates of tested microbes measured with qPCR suggest that qPCR may overestimate the surviving microbes. Collectively, PAA showed broad disinfection efficiency (susceptibility: E. faecium > coliphages > spores). For E. faecium and spores, ATP bioluminescence was substantially faster (∼5 min) than culture-based method (>24 h) and qPCR (2-3 h). This study suggests PAA as an effective alternative to inactivate broad types of microbial contaminants in water. Together with the use of rapid detection methods, this approach can be useful for urgent situations when timely response is needed for ensuring water quality.

  16. Allergic enteritis in children

    PubMed Central

    Czerwionka-Szaflarska, Mieczysława; Gawryjołek, Julia

    2017-01-01

    The gastrointestinal form of food allergy is very common in children. The most frequently observed types are allergic proctitis and proctocolitis. In most cases the symptoms subside within the first 2 months of life. The babies seem healthy, and the only abnormality is a small amount of blood in stool. Symptoms can also include small intestine inflammation and colitis. Patients may present with irritability, abdominal pain, flatulence, colic, postprandial vomiting, chronic diarrhoea, and hindered physical development. The diagnosis of allergic enteritis is based on the clinical examination and the results of additional tests including an endoscopy of the lower digestive tract with histopathological assessment. Cow’s milk proteins are the most common nutrition proteins responsible for the development of the symptoms of allergic enteritis. The most essential method of treating allergic enteritis is the elimination diet. The symptoms should subside within 1–2 weeks from the beginning of the diet. PMID:28337229

  17. Inactivation of hepatitis A virus and indicator organisms in water by free chlorine residuals.

    PubMed

    Grabow, W O; Gauss-Müller, V; Prozesky, O W; Deinhardt, F

    1983-09-01

    Hepatitis A virus (HAV) and selected indicator organisms were mixed together in chlorine-demand-free buffers at pH 6, 8, or 10 and exposed to free chlorine residuals, and the survival kinetics of individual organisms were compared. HAV was enumerated by a most-probable-number dilution assay, using PLC/PRF/5 liver cells for propagation of the virus and radioimmunoassay for its detection. At all pH levels, HAV was more sensitive than Mycobacterium fortuitum, coliphage V1 (representing a type of phage common in some sewage-polluted waters), and poliovirus type 2. Under certain conditions, HAV was more resistant than Escherichia coli, Streptococcus faecalis, coliphage MS2, and reovirus type 3. It was always more resistant than SA-11 rotavirus. Evidence is presented that conditions generally specified for the chlorine disinfection of drinking-water supplies will also successfully inactivate HAV and that HAV inactivation by free chlorine residuals can reliably be monitored by practical indicator systems consisting of appropriate combinations of suitable indicators such as coliform and acid-fast bacteria, coliphages, the standard plate count, and fecal streptococci.

  18. ENFit Enteral Nutrition Connectors.

    PubMed

    Guenter, Peggi; Lyman, Beth

    2016-12-01

    New enteral connectors are now available based on the development of standards using the International Organization of Standardization process to prevent misconnections between systems that should not connect. Enteral devices with the new patient access connectors, called ENFit, are being now introduced for the purpose of improving patient safety. Transitioning to these new connectors poses benefits and challenges for facilities or agencies implementing these new devices. Information from appropriate resources should be sought by clinicians who need to partner with their suppliers and clinical organizations to see how best to meet these challenges.

  19. Enteric Redmouth Disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Yersinia ruckeri, the causative agent of Enteric Redmouth Disease (ERM), is a disease of salmonid fish species that is endemic in areas of the world where salmonids are intensively cultured. The disease causes a chronic to acute hemorrhagic septicemia which can lead to high rates of mortality partic...

  20. Enteric bacteria mandibular osteomyelitis.

    PubMed

    Scolozzi, Paolo; Lombardi, Tommaso; Edney, Timothy; Jaques, Bertrand

    2005-06-01

    Osteomyelitis of the mandible is a relatively rare inflammatory disease that usually stems from the odontogenic polymicrobial flora of the oral cavity. We are reporting 2 unusual cases of mandibular osteomyelitis resulting from enteric bacteria infection. In one patient, abundant clinical evidence suggested a diagnosis of a chronic factitious disease, whereas in the second patient no obvious etiology was found.

  1. Sunlight inactivation of MS2 coliphage in the absence of photosensitizers: modeling the endogenous inactivation rate using a photoaction spectrum.

    PubMed

    Nguyen, Mi T; Silverman, Andrea I; Nelson, Kara L

    2014-04-01

    The endogenous sunlight inactivation rates of MS2 coliphage in photosensitizer-free water were measured (kobs) under different light conditions and compared to modeled inactivation rates (kmod) computed using a previously published action spectrum. Experiments were conducted under simulated and natural sunlight. There was generally good agreement between modeled and observed MS2 sunlight inactivation rates in the summer and winter, suggesting that the action spectrum can be used to predict changes in the inactivation rate caused by diurnal and seasonal changes in natural sunlight irradiance. However, we show that a major source of uncertainty in the predictions is the ability to accurately measure or model the comparatively weak and highly variable solar irradiance between 280 and 300 nm, a range to which the inactivation rate is very sensitive. The action spectrum was also used to predict the endogenous inactivation rates of MS2 at different depths in a column of strongly humic-colored [i.e., solar ultraviolet (UV)-attenuating] wetland water under simulated sunlight; we observed fairly good agreement between kobs and kmod, suggesting that the action spectrum can be used to estimate the decrease in the endogenous inactivation rate caused by spectrally selective sunlight attenuation in the water column.

  2. Fibre and enteral nutrition.

    PubMed Central

    Silk, D B

    1989-01-01

    The recent launch of a number of fibre enriched polymeric diet in the United States and Europe has stimulated considerable interest in the topic of fibre and enteral nutrition, and several commercial concerns appear to be under considerable pressures from their consumers to produce similar products. As a means of identifying areas of potential application of fibre to enteral nutrition some of the recent knowledge gained about the physical properties of dietary fibre and the processes involved in the intestinal assimilation of fibre has been reviewed. Two areas of interest are identifiable. The first relates to the bulking properties of fibre and the application of this to the regulation of bowel function in enterally fed patients. It is clear from the clinical studies that have been reviewed that there remains a paucity of controlled data, and a great deal more research is needed before widespread use of fibre supplemented diets can be supported. Perhaps of greater interest academically is the potentially beneficial effects that appear to be exerted by the VFA's, liberated as a consequence of colonic bacterial fermentation of fibre, on morphology and function of ileal and colonic mucosa. Although there are a number of potential applications of fibre supplemented enteral diets in this area, more research is required before any firm recommendations can be made about recommending their use. The one exception concerns patients with the nutritionally inadequate short bowel syndrome. There does seem to be sufficient experimental evidence to suggest that clinical studies should be commenced using a pectin supplemented predigested 'elemental' diet in these patients. Overall therefore, one is forced to conclude that the increasing interest and use of fibre supplemented enteral diets is being driven more by market than scientific forces. Nevertheless, the promotion of these diets has already provided a powerful stimulus to the scientific community, and it remains entirely

  3. Enteric Viral Surrogate Reduction by Chitosan.

    PubMed

    Davis, Robert; Zivanovic, Svetlana; Davidson, P Michael; D'Souza, Doris H

    2015-12-01

    Enteric viruses are a major problem in the food industry, especially as human noroviruses are the leading cause of nonbacterial gastroenteritis. Chitosan is known to be effective against some enteric viral surrogates, but more detailed studies are needed to determine the precise application variables. The main objective of this work was to determine the effect of increasing chitosan concentration (0.7-1.5% w/v) on the cultivable enteric viral surrogates, feline calicivirus (FCV-F9), murine norovirus (MNV-1), and bacteriophages (MS2 and phiX174) at 37 °C. Two chitosans (53 and 222 kDa) were dissolved in water (53 kDa) or 1% acetic acid (222 KDa) at 0.7-1.5%, and were then mixed with each virus to obtain a titer of ~5 log plaque-forming units (PFU)/mL. These mixtures were incubated for 3 h at 37 °C. Controls included untreated viruses in phosphate-buffered saline and viruses were enumerated by plaque assays. The 53 kDa chitosan at the concentrations tested reduced FCV-F9, MNV-1, MS2, and phi X174 by 2.6-2.9, 0.1-0.4, 2.6-2.8, and 0.7-0.9 log PFU/mL, respectively, while reduction by 222 kDa chitosan was 2.2-2.4, 0.8-1.0, 2.6-5.2, and 0.5-0.8 log PFU/mL, respectively. The 222 kDa chitosan at 1 and 0.7% w/v in acetic acid (pH 4.5) caused the greatest reductions of MS2 by 5.2 logs and 2.6 logs, respectively. Overall, chitosan treatments showed the greatest reduction of MS2, followed by FCV-F9, phi X174, and MNV-1. These two chitosans may contribute to the reduction of enteric viruses at the concentrations tested but would require use of other hurdles to eliminate food borne viruses.

  4. Viral enteritis of calves.

    PubMed

    Woode, G N; Bridger, J C

    1975-01-25

    Calf diarrhoea is an important cause of economic loss. The aetiology of the disease is considered with particular reference to the role of viruses. Although many microbial organisms have been associated with the disease, there is doubt concerning their true role as causative agents. Two viruses, neonatal calf diarrhoea reovirus-life agent and calf coronavirus, have been discovered recently which appear to play an important role in many cases of diarrhoea and the evidence for considering them to be primary causes of the disease is discussed.

  5. Enteral nutrition by tube.

    PubMed

    Armstrong, P J; Hand, M S; Frederick, G S

    1990-01-01

    When oral intake is unsatisfactory or contraindicated, maintenance of nutrition by tube feeding is an alternative to the parenteral route. A large volume of research data supports the decision to use the enteral route whenever possible. Entry of food into the alimentary tract is a stimulus to structural and functional maintenance of that tract. Enteral nutrition can be given via indwelling nasoesophageal, pharyngostomy, esophagostomy, percutaneous or surgical gastrostomy, or enterostomy tube. Use of an appropriate catheter, familiarity with the technique used, and careful patient selection and monitoring are important factors in successful tube feeding. Blenderized pet food diets should be fed whenever possible; commercially available liquid diets provide an alternative when tube caliber or patient factors preclude the use of blenderized foods.

  6. Pathogenic characteristics of persistent feline enteric coronavirus infection in cats.

    PubMed

    Vogel, Liesbeth; Van der Lubben, Mariken; te Lintelo, Eddie G; Bekker, Cornelis P J; Geerts, Tamara; Schuijff, Leontine S; Grinwis, Guy C M; Egberink, Herman F; Rottier, Peter J M

    2010-01-01

    Feline coronaviruses (FCoV) comprise two biotypes: feline enteric coronaviruses (FECV) and feline infectious peritonitis viruses (FIPV). FECV is associated with asymptomatic persistent enteric infections, while FIPV causes feline infectious peritonitis (FIP), a usually fatal systemic disease in domestic cats and some wild Felidae. FIPV arises from FECV by mutation. FCoV also occur in two serotypes, I and II, of which the serotype I viruses are by far the most prevalent in the field. Yet, most of our knowledge about FCoV infections relates to serotype II viruses, particularly about the FIPV, mainly because type I viruses grow poorly in cell culture. Hence, the aim of the present work was the detailed study of the epidemiologically most relevant viruses, the avirulent serotype I viruses. Kittens were inoculated oronasally with different doses of two independent FECV field strains, UCD and RM. Persistent infection could be reproducibly established. The patterns of clinical symptoms, faecal virus shedding and seroconversion were monitored for up to 10 weeks revealing subtle but reproducible differences between the two viruses. Faecal virus, i.e. genomic RNA, was detected during persistent FECV infection only in the large intestine, downstream of the appendix, and could occasionally be observed also in the blood. The implications of our results, particularly our insights into the persistently infected state, are discussed.

  7. Mycoplasma viruses.

    PubMed

    Maniloff, J

    1988-01-01

    Unlike bacterial viruses that infect cells bounded by a cell wall, mycoplasma viruses have evolved to enter and propagate in mycoplasma cells bounded only by a single lipid-protein cell membrane. In addition, mycoplasmas have the smallest amount of genetic information of any known cells, so their complexity is constrained by a limited genetic coding capacity. As a consequence of these host cell differences, mycoplasma viruses have been found to have a variety of structures and replication strategies which are different from those of the bacterial viruses. This article is a critical review of mycoplasma viruses infecting the genera Acholeplasma, Spiroplasma, and Mycoplasma; included are data on classification, morphology and structure, biological and physical properties, chemical composition, and productive and lysogenic replication cycles.

  8. Characterization of the helper proteins for the assembly of tail fibers of coliphages T4 and lambda.

    PubMed Central

    Hashemolhosseini, S; Stierhof, Y D; Hindennach, I; Henning, U

    1996-01-01

    Assembly of tail fibers of coliphage T4 requires the action of helper proteins. In the absence of one of these, protein 38 (p38), p37, constituting the distal part of the long tail fiber, fails to oligomerize. In the absence of the other, p57, p34 (another component of the long tail fiber), p37, and p12 (the subunit of the short tail fiber) remain unassembled. p38 can be replaced by the Tfa (tail fiber assembly) protein (pTfa) of phage lambda, which has the advantage of remaining soluble even when produced in massive amounts. The mechanisms of action of the helpers are unknown. As a first step towards elucidation of these mechanisms, p57 and pTfa have been purified to homogeneity and have been crystallized. The identity of gene 57 (g57), not known with certainty previously, has been established. The 79-residue protein p57 represents a very exotic polypeptide. It is oligomeric and acidic (an excess of nine negative charges). It does not contain Phe, Trp, Tyr, His, Pro, and Cys. Only 25 N-terminal residues were still able to complement a g57 amber mutant, although with a reduced efficiency. In cells overproducing the protein, it assumed a quasi-crystalline structure in the form of highly ordered fibers. They traversed the cells longitudinally (and thus blocked cell division) with a diameter approaching that of the cell and with a hexagonal appearance. The 194-residue pTfa is also acidic (an excess of 13 negative charges) and is likely to be dimeric. PMID:8892827

  9. Impact of riparian zone protection from cattle on nutrient, bacteria, F-coliphage, and loading of an intermittent stream.

    PubMed

    Sunohara, M D; Topp, E; Wilkes, G; Gottschall, N; Neumann, N; Ruecker, N; Jones, T H; Edge, T A; Marti, R; Lapen, D R

    2012-01-01

    This 5-yr study compared, via an upstream-downstream experimental design, nutrient and microbial water quality of an intermittent stream running through a small pasture (∼2.5 animals ha) where cattle are restricted from the riparian zone (restricted cattle access [RCA]) and where cattle have unrestricted access to the stream (unrestricted cattle access [URCA]). Fencing in the RCA excluded pasturing cattle to within ∼3 to 5 m of the stream. Approximately 88% (26/32) of all comparisons of mean contaminant load reduction for lower, higher, and all stream flow conditions during the 5-yr study indicated net contaminant load reductions in the RCA; for the URCA, this percentage was 38% (12/32). For all flow conditions, mean percent load reductions in the RCA for nutrients and bacteria plus F-coliphage were 24 and 23%, respectively. These respective percentages for the URCA were -9 and -57% (positive values are reductions; negative values are increases). However, potentially as a result of protected wildlife habitat in the RCA, the mean percent load reduction for for "all flow" was -321% for the RCA and 60% for the URCA; for , these respective percentages were -209% (RCA) and 73% (URCA). For "all flow" situations, mean load reductions for the RCA were significantly greater ( < 0.1) than those from the URCA for NH-N, dissolved reactive phosphorus (DRP), total coliform, , and . For "high flow" situations, mean load reductions were significantly greater for the RCA for DRP, total coliform, and . For "low flow" conditions, significantly greater mean load reductions were in favor of the RCA for DRP, total P, total coliforms, fecal coliforms, , and . In no case were mean pollutant loads in the URCA significantly higher than RCA pollutant loads. Restricting pasturing livestock to within 3 to 5 m of intermittent streams can improve water quality; however, water quality impairment can occur if livestock have unrestricted access to a stream.

  10. Use of Aqueous Silver To Enhance Inactivation of Coliphage MS-2 by UV Disinfection

    PubMed Central

    Butkus, Michael A.; Labare, Michael P.; Starke, Jeffrey A.; Moon, King; Talbot, Mark

    2004-01-01

    A synergistic effect between silver and UV radiation has been observed that can appreciably enhance the effectiveness of UV radiation for inactivation of viruses. At a fluence of ca. 40 mJ/cm2, the synergistic effect between silver and UV was observed at silver concentrations as low as 10 μg/liter (P < 0.0615). At the same fluence, an MS-2 inactivation of ca. 3.5 logs (99.97%) was achieved at a silver concentration of 0.1 mg/liter, a significant improvement (P < 0.0001) over the ca. 1.8-log (98.42%) inactivation of MS-2 at ca. 40 mJ/cm2 in the absence of silver. Modified Chick-Watson kinetics were used to model the synergistic effect of silver and UV radiation. For an MS-2 inactivation of 4 logs (99.99%), the coefficient of dilution (n) was determined to be 0.31, which suggests that changes in fluence have a greater influence on inactivation than does a proportionate change in silver concentration. PMID:15128542

  11. Total coliphages removal by activated sludge process and their morphological diversity by transmission electron microscopy.

    PubMed

    Jebri, Sihem; Hmaied, Fatma; Yahya, Mariem; Ben Ammar, Aouatef; Hamdi, Moktar

    This study was conducted to isolate phages in treated sewage collected from wastewater treatment plant, and explore their morphological diversity by transmission electron microscopy (TEM). Fates of total bacteriophages and their reduction by biological treatment were also assayed. Phages were isolated using the plaque assay then negatively stained and observed by electron microscope. Electron micrographs showed different types of phages with different shapes and sizes. The majority of viruses found in treated sewage ranged from 30 to 100 nm in capsid diameter. Many of them were tailed, belonging to Siphoviridae, Myoviridae and Podoviridae families. Non-tailed phage particles were also found at a low rate, presumably belonging to Leviviridae or Microviridae families. This study shows the diversity and the abundance of bacteriophages in wastewater after biological treatment. Their persistence in wastewater reused in agriculture should raise concerns about their potential role in controlling bacterial populations in the environment. They should be also included in water treatment quality controlling guidelines as fecal and viral indicators.

  12. An introduction to food and waterborne viruses: diseases, transmission, outbreaks, detection and control

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Enteric viruses are the number one cause of foodborne illness throughout the world. In addition to foods, contaminated drinking water is another major cause of enteric viral illness. Among the enteric viruses are the noroviruses, hepatitis A and E viruses, enteric adenoviruses, rotavirus, and astro...

  13. [Enteral tube feeding].

    PubMed

    Haller, Alois

    2014-03-01

    Tube feeding is an integral part of medical therapies, and can be easily managed also in the outpatient setting. Tube feeding by the stomach or small intestine with nasogastral or nasojejunal tubes is common in clinical practice. Long-term nutrition is usually provided through a permanent tube, i. e. a percutaneous endoscopic gastrostomy (PEG). Modern portable nutrition pumps are used to cover the patient's nutritional needs. Enteral nutrition is always indicated if patients can not or should not eat or if nutritional requirements cannot be covered within 3 days after an intervention, e. g. after abdominal surgery. Industrially produced tube feedings with defined substrate concentrations are being used; different compositions of nutrients, such as glutamine fish oil etc., are used dependent on the the condition of the patient. Enteral nutrition may be associated with complications of the tube, e. g. dislocation, malposition or obstruction, as well as the feeding itself, e. g.hyperglycaemia, electrolyte disturbances, refeeding syndrome diarrhea or aspiration). However, the benefit of tube feeding usually exceeds the potential harm substantially.

  14. John Glenn Entering Friendship 7

    NASA Technical Reports Server (NTRS)

    1962-01-01

    Overall view of astronaut John Glenn, Jr., as he enters into the spacecraft Friendship 7 prior to MA-6 launch operations at Launch Complex 14. Astronaut Glenn is entering his spacecraft to begin the first American manned Earth orbital mission.

  15. Enteral nutrition - child - managing problems

    MedlinePlus

    ... page: //medlineplus.gov/ency/patientinstructions/000164.htm Enteral nutrition - child - managing problems To use the sharing features ... trouble breathing, call 911. References Mcclave SA. Enteral nutrition. In: Goldman L, Schafer AI, eds. Goldman-Cecil ...

  16. HIV, opiates, and enteric neuron dysfunction.

    PubMed

    Galligan, J J

    2015-04-01

    Human immune deficient virus (HIV) is an immunosuppressive virus that targets CD4(+) T-lymphocytes. HIV infections cause increased susceptibility to opportunistic infections and cancer. HIV infection can also alter central nervous system (CNS) function causing cognitive impairment. HIV does not infect neurons but it does infect astrocytes and microglia in the CNS. HIV can also infect enteric glia initiating an intestinal inflammatory response which causes enteric neural injury and gut dysfunction. Part of the inflammatory response is HIV induced production of proteins including, Transactivator of transcription (Tat) which contribute to neuronal injury after release from HIV infected glial cells. A risk factor for HIV infection is intravenous drug use with contaminated needles and chronic opiate use can exacerbate neural injury in the nervous system. While most research focuses on the actions of Tat and other HIV related proteins and opiates on the brain, recent data indicate that Tat can cause intestinal inflammation and disruption of enteric neuron function, including alteration of Na(+) channel activity and action potential generation. A paper published in this issue of Neurogastroenterology and Motility extends these findings by identifying an interaction between Tat and morphine on enteric neuron Na(+) channels and on intestinal motility in vivo using a Tat expressing transgenic mouse model. These new data show that Tat protein can enhance the inhibitory actions of morphine on action potential generation and propulsive motility. These findings are important to our understanding of how HIV causes diarrhea in infected patients and for the use of opioid drugs to treat HIV-induced diarrhea.

  17. [Fiber and enteral nutrition].

    PubMed

    Gómez Candela, C; de Cos Blanco, A I; Iglesias Rosado, C

    2002-01-01

    Dietary fibre is a mixture of various substances and is essential for maintaining appropriate intestinal functionality and it is currently considered to be a necessary part of a healthy diet. Current recommendations for fibre consumption by adults range from 20 to 35 g/day. Enteral nutrition is an emerging therapeutic variation in both hospital and domestic settings. To a great extent, this development has been made possible thanks to the design of new formulas that adapt better and better to the clinicla conditions or our patients. The type of fibre used in these preparations varies greatly. Some have only one source of fibre while others use differnet combinations. There are currently 32 formulas available on the Spanish market, without counting the modules or specific preparations of individual types of fibre. Despite the enormous advances in the knowledge of the beneficial effects of fibre, the fact of the matter is that enteral nutrition that we routinely prescribe in normal clinical practice does not contain fibre. The are several explanations for this, perhaps the most plausible is that these formulas may lead to problems in their administration and tolerance. It is necessary to choose the correct calibre of catheter and define the best infusion method and timing. Another difficulty may be the gastrointestinal tolerance of the formulas containing fibre. No large-scale problems of intolerance have however been described in healthy volunteers nor in patients with acute or chronic pathologies, although it is of fundamental importance to monitor the rhythm of depositions in all patients with enteral nutrition (EN) and ensure proper intake of liquids, which would also be useful to prevent occlusion of the catheter. The theoretical benefits of EN with fibre with a view to maintaining or improving normal intestinal structure and function are very varied. Nonetheless, it has noit yet been possible to prove many of these effects in controlled clinical trials. At the

  18. Entering the Anthropocene

    NASA Astrophysics Data System (ADS)

    Vince, Gaia

    2016-04-01

    There is growing evidence that we are now entering a new geological age defined by human influence on the planet, the Anthropocene. Millions of years from now, a stripe in the accumulated layers of rock on Earth's surface will reveal our human fingerprint just as we can see evidence of dinosaurs in rocks of the Jurassic, or the explosion of life that marks the Cambrian. There is now no part of the planet untouched by human influence. The realisation that we wield such planetary power requires a quite extraordinary shift in perception, fundamentally toppling the scientific, cultural and religious philosophies that define our place in the world. This session explores these issues and examines our new relationship with nature now that we so strongly influence the biosphere. And this session will look at what the impacts of our planetary changes mean for us, and how we might deal with the consequences of the Anthropocene we have created.

  19. Rapid determination of the presence of enteric bacteria in water.

    PubMed

    Kenard, R P; Valentine, R S

    1974-03-01

    A rapid and sensitive method is described for the detection of bacteria in water and various other natural substrates by the isolation of specific bacteriophage. By the addition of large numbers of the organism in question to the sample, the presence of virulent bacteriophage can be demonstrated in as little as 6 to 8 h. Fecal coliform, total coliform, and total coliphage counts were determined for over 150 water samples from several geographical areas over a period of 2 years. Computer analysis of the data shows a high degree of correlation between fecal coliforms and the coliphage present in the samples. With a high correlation coefficient between fecal coliform and coliphage counts, predictions of the fecal coliforms may be made by enumeration of the phage.

  20. Bacterial indicator occurrence and the use of an F+ specific RNA coliphage assay to identify fecal sources in Homosassa Springs, Florida

    USGS Publications Warehouse

    Griffin, Dale W.; Stokes, Rodger; Rose, J.B.; Paul, J.H.

    2000-01-01

    A microbiological water quality study of Homosassa Springs State Wildlife Park (HSSWP) and surrounding areas was undertaken. Samples were collected in November of 1997 (seven sites) and again in November of 1998 (nine sites). Fecal bacterial concentrations (total and fecal coliforms, Clostridium perfringens, and enterococci) were measured as relative indicators of fecal contamination. F+-specific coliphage genotyping was performed to determine the source of fecal contamination at the study sites. Bacterial levels were considerably higher at most sites in the 1997 sampling compared to the 1998 sampling, probably because of the greater rainfall that year. In November of 1997, 2 of the 7 sites were in violation of all indicator standards and guidance levels. In November of 1998, 1 of 9 sites was in violation of all indicator standard and guidance levels. The highest concentrations of all fecal indicators were found at a station downstream of the animal holding pens in HSSWP. The lowest levels of indicators were found at the Homosassa Main Spring vent. Levels of fecal indicators downstream of HSSWP (near the point of confluence with the river) were equivalent to those found in the Southeastern Fork and areas upstream of the park influences. F+ specific RNA coliphage analysis indicated that fecal contamination at all sites that tested positive was from animal sources (mammals and birds). These results suggest that animal (indigenous and those in HSSWP) and not human sources influenced microbial water quality in the area of Homosassa River covered by this study.

  1. Enteric Immunization of Mice Against Influenza with Recombinant Vaccinia

    NASA Astrophysics Data System (ADS)

    Meitin, Catherine A.; Bender, Bradley S.; Small, Parker A., Jr.

    1994-11-01

    Intrajejunal administration to mice of a recombinant vaccinia virus containing the influenza virus hemagglutinin gene induced IgA antibody in nasal, gut, and vaginal secretions. It also induced IgG antibody in serum and cell-mediated immunity. The immunization provided significant protection against an influenza virus challenge. This work suggests that enteric-coated recombinant vaccinia could be an orally administered, inexpensive, multivalent, temperature-stable, safe, and effective vaccine for children that could be particularly useful in developing nations, where multiple injections are not easily administered. Oral administration of vaccines should also reduce children's fear of shots at the doctor's office.

  2. Meta-Analysis of the Reduction of Norovirus and Male-Specific Coliphage Concentrations in Wastewater Treatment Plants

    PubMed Central

    Pouillot, Régis; Van Doren, Jane M.; Woods, Jacquelina; Plante, Daniel; Smith, Mark; Goblick, Gregory; Roberts, Christopher; Locas, Annie; Hajen, Walter; Stobo, Jeffrey; White, John; Holtzman, Jennifer; Buenaventura, Enrico; Burkhardt, William; Catford, Angela; Edwards, Robyn; DePaola, Angelo

    2015-01-01

    Human norovirus (NoV) is the leading cause of foodborne illness in the United States and Canada. Wastewater treatment plant (WWTP) effluents impacting bivalve mollusk-growing areas are potential sources of NoV contamination. We have developed a meta-analysis that evaluates WWTP influent concentrations and log10 reductions of NoV genotype I (NoV GI; in numbers of genome copies per liter [gc/liter]), NoV genotype II (NoV GII; in gc/liter), and male-specific coliphage (MSC; in number of PFU per liter), a proposed viral surrogate for NoV. The meta-analysis included relevant data (2,943 measurements) reported in the scientific literature through September 2013 and previously unpublished surveillance data from the United States and Canada. Model results indicated that the mean WWTP influent concentration of NoV GII (3.9 log10 gc/liter; 95% credible interval [CI], 3.5, 4.3 log10 gc/liter) is larger than the value for NoV GI (1.5 log10 gc/liter; 95% CI, 0.4, 2.4 log10 gc/liter), with large variations occurring from one WWTP to another. For WWTPs with mechanical systems and chlorine disinfection, mean log10 reductions were −2.4 log10 gc/liter (95% CI, −3.9, −1.1 log10 gc/liter) for NoV GI, −2.7 log10 gc/liter (95% CI, −3.6, −1.9 log10 gc/liter) for NoV GII, and −2.9 log10 PFU per liter (95% CI, −3.4, −2.4 log10 PFU per liter) for MSCs. Comparable values for WWTPs with lagoon systems and chlorine disinfection were −1.4 log10 gc/liter (95% CI, −3.3, 0.5 log10 gc/liter) for NoV GI, −1.7 log10 gc/liter (95% CI, −3.1, −0.3 log10 gc/liter) for NoV GII, and −3.6 log10 PFU per liter (95% CI, −4.8, −2.4 PFU per liter) for MSCs. Within WWTPs, correlations exist between mean NoV GI and NoV GII influent concentrations and between the mean log10 reduction in NoV GII and the mean log10 reduction in MSCs. PMID:25934626

  3. Weather and environmental factors associated with F+ coliphages and fecal indicator bacteria in beach sand at two recreational marine beaches

    EPA Science Inventory

    Studies have demonstrated that fecal indicator bacteria (FIB) and pathogens may be present in beach sand and suggest an increased risk of enteric illness among beachgoers contacting sand. During the 2007 National Epidemiological and Environmental Assessment of Recreational (NEEAR...

  4. Microscopic enteritis: Bucharest consensus.

    PubMed

    Rostami, Kamran; Aldulaimi, David; Holmes, Geoffrey; Johnson, Matt W; Robert, Marie; Srivastava, Amitabh; Fléjou, Jean-François; Sanders, David S; Volta, Umberto; Derakhshan, Mohammad H; Going, James J; Becheanu, Gabriel; Catassi, Carlo; Danciu, Mihai; Materacki, Luke; Ghafarzadegan, Kamran; Ishaq, Sauid; Rostami-Nejad, Mohammad; Peña, A Salvador; Bassotti, Gabrio; Marsh, Michael N; Villanacci, Vincenzo

    2015-03-07

    Microscopic enteritis (ME) is an inflammatory condition of the small bowel that leads to gastrointestinal symptoms, nutrient and micronutrient deficiency. It is characterised by microscopic or sub-microscopic abnormalities such as microvillus changes and enterocytic alterations in the absence of definite macroscopic changes using standard modern endoscopy. This work recognises a need to characterize disorders with microscopic and submicroscopic features, currently regarded as functional or non-specific entities, to obtain further understanding of their clinical relevance. The consensus working party reviewed statements about the aetiology, diagnosis and symptoms associated with ME and proposes an algorithm for its investigation and treatment. Following the 5(th) International Course in Digestive Pathology in Bucharest in November 2012, an international group of 21 interested pathologists and gastroenterologists formed a working party with a view to formulating a consensus statement on ME. A five-step agreement scale (from strong agreement to strong disagreement) was used to score 21 statements, independently. There was strong agreement on all statements about ME histology (95%-100%). Statements concerning diagnosis achieved 85% to 100% agreement. A statement on the management of ME elicited agreement from the lowest rate (60%) up to 100%. The remaining two categories showed general agreement between experts on clinical presentation (75%-95%) and pathogenesis (80%-90%) of ME. There was strong agreement on the histological definition of ME. Weaker agreement on management indicates a need for further investigations, better definitions and clinical trials to produce quality guidelines for management. This ME consensus is a step toward greater recognition of a significant entity affecting symptomatic patients previously labelled as non-specific or functional enteropathy.

  5. Detection of enteric pathogens in Turkey flocks affected with severe enteritis, in Brazil.

    PubMed

    Moura-Alvarez, Joelma; Nuñez, Luis F N; Astolfi-Ferreira, Claudete S; Knöbl, Terezinha; Chacón, Jorge L; Moreno, Andrea M; Jones, Richard C; Ferreira, Antonio J Piantino

    2014-08-01

    Twenty-two flocks of turkeys affected by enteric problems, with ages between 10 and 104 days and located in the Southern region of Brazil, were surveyed for turkey by PCR for turkey astrovirus type 2 (TAstV-2), turkey coronavirus (TCoV), hemorrhagic enteritis virus (HEV), rotavirus, reovirus, Salmonella spp., and Lawsonia intracellularis (Li) infections. Eleven profiles of pathogen combination were observed. The most frequently encountered pathogen combinations were TCoV-Li, followed by TCoV-TAstV-2-Li, TCoV-TastV-2. Only TCoV was detected as the sole pathogen in three flocks. Eight and 19 flocks of the 22 were positive for TAstV-2 and TCoV, respectively. Six were positive for Salmonella spp. and L. intracellularis was detected in 12 turkey flocks. Reovirus and HEV were not detected in this survey. These results throw new light on the multiple etiology of enteritis in turkeys. The implications of these findings and their correlation with the clinical signs are comprehensively discussed, illustrating the complexity of the enteric diseases.

  6. Partial Genome Sequence Analysis of Parvoviruses Associated with Enteric Disease in Poultry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Poult Enteritis Mortality Syndrome (PEMS) of turkeys and Runting-Stunting Syndrome (RSS) of chickens are significant viral enteric diseases of poultry. Although a number of different viruses, including avian reoviruses, rotaviruses, astroviruses and coronaviruses, have been isolated from the intesti...

  7. Partial genome sequence analysis of parvoviruses associated with enteric disease of poultry.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Poult Enteritis Mortality Syndrome (PEMS) of turkeys and Runting-Stunting Syndrome (RSS) of chickens are significant viral enteric diseases of poultry. Although numerous different viruses have been isolated from the intestinal contents of affected poultry flocks, including avian reoviruses, rotaviru...

  8. Comparative metagenomic analyses of the poultry enteric virome: implications for diagnostics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previous comparative metagenomic analyses of the viral communities present in the poultry gut have revealed novel enteric viruses that may affect the overall performance of birds or that may play roles in disease syndromes such as Runting-Stunting Syndrome (RSS) or Poult Enteritis Complex (PEC). Ou...

  9. Inactivation of Viruses and Bacteriophages as Models for Swine Hepatitis E Virus in Food Matrices.

    PubMed

    Emmoth, Eva; Rovira, Jordi; Rajkovic, Andreja; Corcuera, Elena; Wilches Pérez, Diego; Dergel, Irene; Ottoson, Jakob R; Widén, Frederik

    2017-03-01

    Hepatitis E virus has been recognised as a food-borne virus hazard in pork products, due to its zoonotic properties. This risk can be reduced by adequate treatment of the food to inactivate food-borne viruses. We used a spectrum of viruses and bacteriophages to evaluate the effect of three food treatments: high pressure processing (HPP), lactic acid (LA) and intense light pulse (ILP) treatments. On swine liver at 400 MPa for 10 min, HPP gave log10 reductions of ≥4.2, ≥5.0 and 3.4 for feline calicivirus (FCV) 2280, FCV wildtype (wt) and murine norovirus 1 (MNV 1), respectively. Escherichia coli coliphage ϕX174 displayed a lower reduction of 1.1, while Escherichia coli coliphage MS2 was unaffected. For ham at 600 MPa, the corresponding reductions were 4.1, 4.4, 2.9, 1.7 and 1.3 log10. LA treatment at 2.2 M gave log10 reductions in the viral spectrum of 0.29-2.1 for swine liver and 0.87-3.1 for ham, with ϕX174 and MNV 1, respectively, as the most stable microorganisms. The ILP treatment gave log10 reductions of 1.6-2.8 for swine liver, 0.97-2.2 for ham and 1.3-2.3 for sausage, at 15-60 J cm(-2), with MS2 as the most stable microorganism. The HPP treatment gave significantly (p < 0.05) greater virus reduction on swine liver than ham for the viruses at equivalent pressure/time combinations. For ILP treatment, reductions on swine liver were significantly (p < 0.05) greater than on ham for all microorganisms. The results presented here could be used in assessments of different strategies to protect consumers against virus contamination and in advice to food producers. Conservative model indicators for the pathogenic viruses could be suggested.

  10. Global occurrence of Torque teno virus in water systems.

    PubMed

    Charest, A J; Plummer, J D; Long, S C; Carducci, A; Verani, M; Sidhu, J P S

    2015-09-01

    Bacterial indicator organisms are used globally to assess the microbiological safety of waters. However, waterborne viral outbreaks have occurred in drinking water systems despite negative bacterial results. Using viral markers may therefore provide more accurate health risk assessment data. In this study, fecal, wastewater, stormwater, surface water (fresh and salt), groundwater, and drinking water samples were analyzed for the presence or concentration of traditional indicators, innovative indicators and viral markers. Samples were obtained in the United States, Italy, and Australia and results compared to those reported for studies conducted in Asia and South America as well. Indicators included total coliforms, Escherichia coli, enterococci, male-specific coliphages, somatic coliphages and microviradae. Viral markers included adenovirus, polyomavirus, and a potential new surrogate, Torque teno virus (TTV). TTV was more frequently found in wastewaters (38-100%) and waters influenced by waste discharges (25%) than in surface waters used as drinking water sources (5%). TTV was also specific to human rather than animal feces. While TTV numbers were strongly correlated to other viral markers in wastewaters, suggesting its utility as a fecal contamination marker, data limitations and TTV presence in treated drinking waters demonstrates that additional research is needed on this potential viral indicator.

  11. DEVELOPMENT OF MULTIPLEX RT-PCR FOR THE DETECTION OF REOVIRUS, HEPATITIS A VIRUS, POLIOVIRUS, NORWALK VIRUS AND ROTAVIRUS

    EPA Science Inventory

    Water sources are often found to be contaminated by enteric viruses. This is a public health concern as food and waterborne outbreaks caused by enteric viruses such as noroviruses, rotaviruses, hepatitis A virus (HAV) and enteroviruses are a common occurrence. All of these viru...

  12. Settling and survival profile of enteric pathogens in the swine effluent for water reuse purpose.

    PubMed

    Fongaro, G; Kunz, A; Magri, M E; Schissi, C D; Viancelli, A; Philippi, L S; Barardi, C R M

    2016-11-01

    The present study evaluated the pathogens persistence and settling profile in swine effluent. We determined the enteric pathogens settling characteristics, their survival and inactivation profile in swine effluent (for water reuse purpose) and in sludge (generated after aerobic treatment - during secondary settling process). The study was performed in laboratorial-scale and in full-scale (manure treatment plant). Enteric viruses and enteric bacteria were used as biomarkers. Results showed that these enteric pathogens were significantly reduced from swine effluent during secondary settling process, and enteric viruses removal was correlated with the suspended solids decantation. The design of secondary settlers can be adapted to improve pathogens removal, by diminishing the solids loading rate per area and time, ending in higher hydraulic retention times.

  13. Fecal indicators in sand, sand contact, and risk of enteric illness among beachgoers

    PubMed Central

    Heaney, Christopher D.; Sams, Elizabeth; Dufour, Alfred P.; Brenner, Kristen P.; Haugland, Richard A.; Chern, Eunice; Wing, Steve; Marshall, Stephen; Love, David C.; Serre, Marc; Noble, Rachel; Wade, Timothy J.

    2011-01-01

    Background Beach sand can harbor fecal indicator organisms and pathogens, but enteric illness risk associated with sand contact remains unclear. Methods In 2007, visitors at two recreational marine beaches were asked on the day of their visit about sand contact. Ten to 12 days later, participants answered questions about health symptoms since the visit. F+ coliphage, Enterococcus, Bacteroidales, fecal Bacteroides, and Clostridium spp. in wet sand were measured using culture and molecular methods. Results We analyzed 144 wet sand samples and completed 4,999 interviews. Adjusted odds ratios (aORs) were computed, comparing those in the highest tertile of fecal indicator exposure with those who reported no sand contact. Among those digging in sand compared with those not digging in sand, a molecular measure of Enterococcus spp. (calibrator cell equivalents/g) in sand was positively associated with gastrointestinal (GI) illness (aOR = 2.0 [95% confidence interval (CI) = 1.2–3.2]) and diarrhea (2.4 [1.4–4.2]). Among those buried in sand, point estimates were greater for GI illness (3.3 [1.3–7.9]) and diarrhea (4.9 [1.8–13]). Positive associations were also observed for culture-based Enterococcus (colony-forming units/g) with GI illness (aOR digging = 1.7 [1.1–2.7]) and diarrhea (2.1 [1.3–3.4]). Associations were not found among non-swimmers with sand exposure. Conclusions We observed a positive relationship between sand contact activities and enteric illness as a function of concentrations of fecal microbial pollution in beach sand. PMID:22157306

  14. Indiscretion enteritis. A Rabelaisian syndrome.

    PubMed

    Robin, E D; Collins, J; Burke, C

    1986-12-01

    A 76-year-old man had small bowel obstruction and organic small bowel disease following a series of bizarre massive gustatory insults that involved food, medications, and mega-mineral-vitamin supplements. Intestinal obstruction required partial small bowel resection. The dietary indiscretions resulted in severe enteritis (indiscretion enteritis). The sequence has been termed a Rabelaisian syndrome after the great French writer and physician, Francois Rabelais, who vividly described bizarre gustatory habits. Gut injury may result from unwise oral intake of various foods and mineral supplements.

  15. Removal of bacteria, protozoa and viruses through a multiple-barrier household water disinfection system.

    PubMed

    Espinosa-García, A C; Díaz-Ávalos, C; Solano-Ortiz, R; Tapia-Palacios, M A; Vázquez-Salvador, N; Espinosa-García, S; Sarmiento-Silva, R E; Mazari-Hiriart, M

    2014-03-01

    Municipal water disinfection systems in some areas are not always able to meet water consumer needs, such as ensuring distributed water quality, because household water management can be a contributing factor in water re-contamination. This fact is related to the storage options that are common in places where water is scarce or is distributed over limited time periods. The aim of this study is to assess the removal capacity of a multiple-barrier water disinfection device for protozoa, bacteria, and viruses. Water samples were taken from households in Mexico City and spiked with a known amount of protozoa (Giardia cyst, Cryptosporidium oocyst), bacteria (Escherichia coli), and viruses (rotavirus, adenovirus, F-specific ribonucleic acid (FRNA) coliphage). Each inoculated sample was processed through a multiple-barrier device. The efficiency of the multiple-barrier device to remove E. coli was close to 100%, and more than 87% of Cryptosporidium oocysts and more than 98% of Giardia cysts were removed. Close to 100% of coliphages were removed, 99.6% of the adenovirus was removed, and the rotavirus was almost totally removed. An effect of site by zone was detected; this observation is important because the water characteristics could indicate the efficiency of the multiple-barrier disinfection device.

  16. Enteral Tube Feeding and Pneumonia

    ERIC Educational Resources Information Center

    Gray, David Sheridan; Kimmel, David

    2006-01-01

    To determine the effects of enteral tube feeding on the incidence of pneumonia, we performed a retrospective review of all clients at our institution who had gastrostomy or jejunostomy tubes placed over a 10-year period. Ninety-three subjects had a history of pneumonia before feeding tube insertion. Eighty had gastrostomy and 13, jejunostomy…

  17. Analysis of multiple enteric viral targets as sewage markers in coral reefs

    USGS Publications Warehouse

    Lipp, Erin K.; Futch, J. Carrie; Griffin, Dale W.

    2007-01-01

    Water and coral mucus samples were collected from throughout the Florida Keys National Marine Sanctuary and the Dry Tortugas for three years and were analyzed for human enteric viruses (enteroviruses, noroviruses, hepatitis A virus and adenoviruses) as conservative markers of human sewage using molecular methods. Of the 100 coral and water samples collected, 40 contained genetic material from one or more human enteric viruses. DNA-based adenoviruses were detected widely, in 37.8% of samples and at 91% of stations, including ‘pristine’ reefs in the Dry Tortugas; however, the detection rate was ⩽12% for the RNA-based enteroviruses and noroviruses (hepatitis A virus was never detected). The disparity between the prevalence of RNA- and DNA-based viruses suggests the need for additional work to determine the utility of adenovirus as marker of human sewage.

  18. A conserved genetic module that encodes the major virion components in both the coliphage T4 and the marine cyanophage S-PM2

    PubMed Central

    Hambly, Emma; Tétart, Francoise; Desplats, Carine; Wilson, William H.; Krisch, Henry M.; Mann, Nicholas H.

    2001-01-01

    Sequence analysis of a 10-kb region of the genome of the marine cyanomyovirus S-PM2 reveals a homology to coliphage T4 that extends as a contiguous block from gene (g)18 to g23. The order of the S-PM2 genes in this region is similar to that of T4, but there are insertions and deletions of small ORFs of unknown function. In T4, g18 codes for the tail sheath, g19, the tail tube, g20, the head portal protein, g21, the prohead core protein, g22, a scaffolding protein, and g23, the major capsid protein. Thus, the entire module that determines the structural components of the phage head and contractile tail is conserved between T4 and this cyanophage. The significant differences in the morphology of these phages must reflect the considerable divergence of the amino acid sequence of their homologous virion proteins, which uniformly exceeds 50%. We suggest that their enormous diversity in the sea could be a result of genetic shuffling between disparate phages mediated by such commonly shared modules. These conserved sequences could facilitate genetic exchange by providing partially homologous substrates for recombination between otherwise divergent phage genomes. Such a mechanism would thus expand the pool of phage genes accessible by recombination to all those phages that share common modules. PMID:11553768

  19. THE APPLICATION OF EMERGING TECHNOLOGIES TO VIRUS DETECTION IN WATER

    EPA Science Inventory

    Human enteric viruses belonging to many different viral genera cause waterborne disease when susceptible individuals are exposed to contaminated drinking and recreational waters. Diseases resulting from infection with these viruses include gastroenteritis, hepatitis, encephali...

  20. Duck viral enteritis in domestic muscovy ducks in Pennsylvania

    USGS Publications Warehouse

    Davison, S.; Converse, K.A.; Hamir, A.N.; Eckroade, R.J.

    1993-01-01

    Duck viral enteritis (DVE) outbreaks occurred at two different locations in Pennsylvania in 1991 and 1992. In the first outbreak, four ducks died out of a group of 30 domestic ducks; in the second outbreak, 65 ducks died out of a group of 114 domestic ducks, and 15 domestic geese died as well. A variety of species of ducks were present on both premises, but only muscovy ducks (Cairina moschata) died from the disease. On necropsy, gross lesions included hepatomegaly with petechial hemorrhages, petechial hemorrhages in the abdominal fat, petechial hemorrhages on the epicardial surface of the heart, and multifocal to coalescing areas of fibrinonecrotic material over the mucosal surface of the trachea, esophagus, intestine, and cloaca. Histologically, the liver had random multifocal areas of necrosis and eosinophilic intranuclear inclusion bodies in hepatocytes. DVE virus was isolated and identified using muscovy duck embryo fibroblast inoculation and virus neutralization. /// En dos sitios diferentes se presentaron brotes de enteritis viral de los patos en el estados de Pensilvania en los a??os 1991 y 1992. En el primer brote, cuatro de un lote de 30 patos murieron mientras que en el segundo brote murieron 65 patos de un lote de 114 patos y 15 gansos. En ambas localidades exist?-a una variedad de especies de patos, sin embargo, s??lamente los patos almizcleros (Cairina moschata) murieron. A la necropsia, las lesiones macrosc??picas incluyeron hepatomegalia con hemorragias petequiales, hemorragias petequiales en la grasa abdominal y en la superficie del epicardio, y ?!reas multifocales o coalescentes de material fibrinonecr??tico sobre la superficie de la mucosa de la tr?!quea, es??fago, intestino y cloaca. Histol??gicamente, el h?-gado mostraba ?!reas multifocales de necrosis y cuerpos de inclusi??n intranucleares eosinof?-licos en los hepatocitos. El virus de la enteritis viral de los patos fue aislado e identificado usando fibroblasto de embriones de pato almizclero

  1. Chronic radiation enteritis and malnutrition.

    PubMed

    Webb, Gwilym James; Brooke, Rachael; De Silva, Aminda Niroshan

    2013-07-01

    Radiation enteritis is defined as the loss of absorptive capacity of the intestine following irradiation, which is most commonly seen after radiotherapy for pelvic and abdominal malignancies. It is divided into acute and chronic forms and usually presents with diarrhea and malabsorption. Malnutrition is a common complication of chronic radiation enteritis (CRE). We reviewed the etiology, prevalence, symptoms, diagnosis and management of CRE and CRE with malnutrition in this article. Functional short bowel syndrome as a cause of malnutrition in CRE is also considered. The diagnostic work-up includes serum markers, endoscopy, cross-sectional imaging and the exclusion of alternative diagnoses such as recurrent malignancy. Management options of CRE include dietary manipulation, anti-motility agents, electrolyte correction, probiotics, parenteral nutrition, surgical resection and small bowel transplantation. Treatment may also be required for coexisting conditions including vitamin B12 deficiency, bile acid malabsorption and depression.

  2. Cyclophosphamide-associated enteritis: A rare association with severe enteritis

    PubMed Central

    Yang, Linda S; Cameron, Karla; Papaluca, Tim; Basnayake, Chamara; Jackett, Louise; McKelvie, Penelope; Goodman, David; Demediuk, Barbara; Bell, Sally J; Thompson, Alexander J

    2016-01-01

    Cyclophosphamide is a potent cytotoxic agent used in many clinical settings. The main risks of cyclophosphamide therapy include hematological disorders, infertility, hemorrhagic cystitis and malignancies. Gastrointestinal side effects reported to date are often non-specific and not severe. We present the first case of a fatal small bowel enteritis and pan-colitis which appears to be associated with cyclophosphamide. We aim to raise the readers’ awareness of this significant adverse event to facilitate clinical suspicion and early recognition in potential future cases. PMID:27818600

  3. Inactivation of adenovirus type 5, rotavirus Wa and male specific coliphage (MS2) in biosolids by lime stabilization.

    PubMed

    Hansen, Jacqueline J; Warden, Paul S; Margolin, Aaron B

    2007-03-01

    The use of lime to reduce or eliminate pathogen content is a cost-effective treatment currently employed in many Class B biosolids production plants in the United States. A bench scale model of lime stabilization was designed to evaluate the survival of adenovirus type 5, rotavirus Wa, and the male specific bacteriophage, MS2, in various matrices. Each virus was initially evaluated independently in a reverse osmosis treated water matrix limed with an aqueous solution of calcium hydroxide for 24-hr at 22 +/- 5 degrees C. In all R/O water trials, adenovirus type 5, rotavirus Wa and MS2 were below detectable levels (<100.5 TCID50/mL and <1 PFU/mL respectively) following 0.1-hr of liming. Adenovirus type 5, rotavirus Wa, and MS2, were inoculated into composted, raw and previously limed matrices, representative of sludge and biosolids, to achieve a final concentration of approximately 104 PFU or TCID50/mL. Each matrix was limed for 24-hr at 22 +/- 5 degrees C and 4 +/- 2 degrees C. In all trials virus was below detectable levels following a 24-hr incubation. The time required for viral inactivation varied depending on the temperature and sample matrix. This research demonstrates reduction of adenovirus type 5, rotavirus Wa, and male-specific bacteriophage, in water, sludge and biosolids matrices following addition of an 8% calcium hydroxide slurry to achieve a pH of 12 for 2-hr reduced to 11.5 for 22-hr by addition of 0.1 N HCl. In these trials, MS2 was a conservative indicator of the efficacy of lime stabilization of adenovirus Type 5 and rotavirus Wa and therefore is proposed as a useful indicator organism.

  4. Glenn Enters his Mercury Capsule

    NASA Technical Reports Server (NTRS)

    1962-01-01

    Astronaut John H. Glenn Jr. enters his Mercury capsule, 'Friendship 7' as he prepares for launch of the Mercury-Atlas rocket. On February 20, 1962 Glenn lifted off into space aboard his Mercury Atlas 6 (MA-6) rocket and became the first American to orbit the Earth. After orbiting the Earth 3 times, Friendship 7 landed in the Atlantic Ocean 4 hours, 55 minutes and 23 seconds later, just East of Grand Turk Island in the Bahamas. Glenn and his capsule were recovered by the Navy Destroyer Noa, 21 minutes after splashdown.

  5. Aerosolization of Respirable Droplets from a Domestic Spa Pool and the Use of MS-2 Coliphage and Pseudomonas aeruginosa as Markers for Legionella pneumophila

    PubMed Central

    Hewitt, Matthew; Stevenson, David; Walker, Jimmy T.; Bennett, Allan M.

    2014-01-01

    Legionnaires' disease can result when droplets or aerosols containing legionella bacteria are inhaled and deposited in the lungs. A number of outbreaks have been associated with the use of a spa pool where aeration, a high water temperature, and a large and variable organic load make disinfectant levels difficult to maintain. Spa pool ownership is increasing, and the aim of this study, using two surrogate organisms (MS-2 coliphage and Pseudomonas aeruginosa [a natural contaminant]), was to assess the potential risk to domestic users when disinfection fails. A representative “entry level” domestic spa pool was installed in an outdoor courtyard. The manufacturer's instructions for spa pool maintenance were not followed. A cyclone sampler was used to sample the aerosols released from the spa pool with and without activation of the air injection system. Samples were taken at increasing heights and distances from the pool. An aerodynamic particle sizer was used to measure the water droplet size distribution at each sample point. When the air injection system was inactivated, neither surrogate organism was recovered from the air. On activation of the air injection system, the mean mass of droplets within the respirable range (10 cm above the water line) was 36.8 μg cm−3. This corresponded to a mean air concentration of P. aeruginosa of 350 CFU m−3. From extrapolation from animal data, the estimated risk of infection from aerosols contaminated with similar concentrations of Legionella pneumophila was 0.76 (males) and 0.65 (females). At 1 m above and/or beyond the pool, the mean aerosol mass decreased to 0.04 μg cm−3 and corresponded to a 100-fold reduction in mean microbial air concentration. The estimated risk of infection at this distance was negligible. PMID:25381233

  6. HIV antibody seroprevalence among prisoners entering the California correctional system.

    PubMed Central

    Singleton, J. A.; Perkins, C. I.; Trachtenberg, A. I.; Hughes, M. J.; Kizer, K. W.; Ascher, M.

    1990-01-01

    A cross-sectional blind study was conducted in the spring of 1988 to estimate the extent of human immunodeficiency virus (HIV) infection among inmates entering the California correctional system. Of the 6,834 inmates receiving entrance physical examinations during the study period, 6,179 (90.4%) had serum tested for the presence of HIV antibodies after routine blood work was completed and personal identifiers were removed. Seroprevalence was 2.5% (95% confidence interval, 2.1% to 3.0%) among the 5,372 men tested and 3.1% (95% confidence interval, 2.1% to 4.5%) among the 807 women tested. Seroprevalence was more than twice as high among men arrested in the San Francisco Bay Area as in those arrested elsewhere in the state. The regional differences in HIV seroprevalence observed among entering inmates mirror infection rates reported among intravenous drug users from the same regions. PMID:2244374

  7. Interferon Gamma as a Biomarker of Exposure to Enteric Viruses

    EPA Science Inventory

    Interferon gamma (IFN-γ) was selected as a biomarker for viral exposure. Twelve-week-old BALB/c mice were intraperitoneally injected with Coxsackievirus B3 or B4 diluted in phosphate-buffered saline (PBS). Control mice were injected with PBS only. Four months after viral infectio...

  8. THE SIGNIFICANCE OF ENTERIC VIRUSES AND WATERBORNE ILLNESS

    EPA Science Inventory

    With growing concern over drinking water safety, considerable attention has been directed towards microbial pathogens in source waters, and the adequacy of current methods used to detect, monitor and treat for these pathogens. The focus has been on bacterial and protozoan pathog...

  9. Airborne enteric bacteria and viruses from spray irrigation with wastewater.

    PubMed

    Teltsch, B; Katzenelson, E

    1978-02-01

    The relationship between bacterial concentrations in wastewater used for spray irrigation and in the air was examined. Aerosolized coliforms were detected when their concentration was 10(3)/ml or more in the wastewater. Relative humidity and solar irradiation appeared to affect viable bacteria in the air; a positive correlation was found between relative humidity and the number of aerosolized bacteria. The correlation between solar irradiation and bacterial level, on the other hand, was negative. During night irrigation, up to 10 times more aerosolized bacteria were detected than with day irrigation. Wind velocity did not play an important role in the survival of aerosolized bacteria. Echovirus 7 was isolated in 4 out of 12 air samples collected 40 m downwind from the sprinkler.

  10. The role of carbohydrates in infection strategies of enteric pathogens.

    PubMed

    Kato, Kentaro; Ishiwa, Akiko

    2015-03-01

    Enteric pathogens cause considerable public health concerns worldwide including tropical regions. Here, we review the roles of carbohydrates in the infection strategies of various enteric pathogens including viruses, bacteria and protozoa, which infect the epithelial lining of the human and animal intestine. At host cell entry, enteric viruses, including norovirus, recognize mainly histo-blood group antigens. At the initial step of bacterial infections, carbohydrates also function as receptors for attachment. Here, we describe the function of carbohydrates in infection by Salmonella enterica and several bacterial species that produce a variety of fimbrial adhesions. During invasion by enteropathogenic protozoa, apicomplexan parasites utilize sialic acids or sulfated glycans. Carbohydrates serve as receptors for infection by these microbes; however, their usage of carbohydrates varies depending on the microbe. On the surface of the mucosal tissues of the gastrointestinal tract, various carbohydrate moieties are present and play a crucial role in infection, representing the site of infection or route of access for most microbes. During the infection and/or invasion process of the microbes, carbohydrates function as receptors for various microbes, but they can also function as a barrier to infection. One approach to develop effective prophylactic and therapeutic antimicrobial agents is to modify the drug structure. Another approach is to modify the mode of inhibition of infection depending on the individual pathogen by using and mimicking the interactions with carbohydrates. In addition, similarities in mode of infection may also be utilized. Our findings will be useful in the development of new drugs for the treatment of enteric pathogens.

  11. Cytomegalovirus enteritis in immunocompetent subjects: a case report and review of the literature.

    PubMed

    Karigane, Daiki; Takaya, Saho; Seki, Yuki; Mastumoto, Yuka; Onose, Akira; Kosakai, Arifumi; Sugaya, Norio; Mori, Takehiko

    2014-05-01

    Cytomegalovirus (CMV) enteritis (or colitis) is generally diagnosed in immunocompromised patients in association with human immunodeficiency virus infection as well as in recipients of solid organ or hematopoietic stem cell transplant. CMV enteritis has been reported only sporadically in immunocompetent individuals. We encountered a 76-year-old woman who developed CMV enteritis without any previously identified immunocompromised states. An extensive literature review of 33 cases of CMV enteritis or colitis diagnosed in immunocompetent individuals, including the present case, revealed that the median age of the patients was 68, the accompanying symptoms were diarrhea (76%), abdominal pain (52%), and hematochezia or melena (27%), and that the outcome was generally favorable, including resolution without any treatment in 24% of the patients. CMV enteritis should be recognized more widely as a disease entity not only in immunocompromised patients but also in immunocompetent individuals, especially in elderly populations.

  12. Enteric spore-forming opportunistic parasites in HIV / AIDS.

    PubMed

    Chawla, Rohit; Ichhpujani, R L

    2011-01-01

    Human Immunodeficiency Virus (HIV) infection causes progressive damage to both limbs of the immune system, which results in a plethora of opportunistic infections. Among the various opportunistic infections, gastrointestinal infections are very common in HIV / Acquired Immunodeficiency Syndrome (AIDS). Opportunistic spore-forming protozoal parasites, namely, Cryptosporidium parvum, Isospora belli, Cyclospora cayetanensis, and Microsporidia, play a major role in causing chronic diarrhea, accompanied with weight loss, in patients with HIV / AIDS. The purpose of this review is to discuss the salient microbiological, clinical, and diagnostic aspects of important enteric spore-forming opportunistic parasites in HIV / AIDS.

  13. Synthesis and processing of structural and intracellular proteins of two enteric coronaviruses

    SciTech Connect

    Sardinia, L.M.

    1985-01-01

    The synthesis and processing of virus-specific proteins of two economically important enteric coronaviruses, bovine enteric coronavirus (BCV) and transmissible gastroenteritis virus (TGEV), were studied at the molecular level. To determine the time of appearance of virus-specific proteins, virus-infected cells were labeled with /sup 35/S-methionine at various times during infection, immunoprecipitated with specific hyperimmune ascitic fluid, and analyzed by SDS-polyacrylamide gel electrophoresis. The peak of BCV protein synthesis was found to be at 12 hours postinfection (hpi). The appearance of all virus-specific protein was coordinated. In contrast, the peak of TGEV protein synthesis was at 8 hpi, but the nucleocapsid proteins was present as early as 4 hpi. Virus-infected cells were treated with tunicamycin to ascertain the types of glycosidic linkages of the glycoproteins. The peplomer proteins of both viruses were sensitive to inhibition by tunicamycin indicating that they possessed N-linked carbohydrates. The matrix protein of TGEV was similarly affected. The matrix protein of BCV, however, was resistant to tunicamycin treatment and, therefore, has O-linked carbohydrates. Only the nucleocapsid protein of both viruses is phosphorylated as detected by radiolabeling with /sup 32/P-orthophosphate. Pulse-chase studies and comparison of intracellular and virion proteins were done to detect precursor-product relationships.

  14. A Potential Mechanism of Virus Persistence within Bivalve Shellfish.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The process of enteric virus bioaccumulation by molluscan shellfish has been widely studied and documented. However, it is not understood why some human enteric viruses, which cannot replicate within shellfish, are selectively retained and remain viable within shellfish tissues for extended periods...

  15. Comparative effectiveness of membrane bioreactors, conventional secondary treatment, and disinfection to remove microorganisms from municipal wastewaters

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Log removals of bacterial indicators, coliphage, and enteric viruses were studied in three membrane bioreactor activated-sludge (MBR) and two conventional secondary activated-sludge municipal wastewater treatment plants during three disinfection seasons (May–Oct.). In total, 73 regular samples were ...

  16. Heartland Virus

    MedlinePlus

    ... Vector-Borne Diseases (DVBD) NCEZID Share Compartir Heartland virus On this Page What is Heartland virus? How ... Do I Need to Know? What is Heartland virus? Heartland virus belongs to a family of viruses ...

  17. EPA METHODS FOR VIRUS DETECTION IN WATER

    EPA Science Inventory

    A number of different types of human enteric viruses cause waterborne outbreaks when individuals are exposed to contaminated drinking and recreational waters. Members of the enterovirus group cause numerous diseases, including gastroenteritis, encephalitis, meningitis, myocard...

  18. A non-enteric adenovirus A12 gastroenteritis outbreak in Rio de Janeiro, Brazil

    PubMed Central

    Portes, Silvana Augusta Rodrigues; Volotão, Eduardo de Mello; Rocha, Monica Simões; Rebelo, Maria Cristina; Xavier, Maria da Penha Trindade Pinheiro; de Assis, Rosane Maria; Rose, Tatiana Lundgren; Miagostovich, Marize Pereira; Leite, José Paulo Gagliardi; Carvalho-Costa, Filipe Anibal

    2016-01-01

    A gastroenteritis outbreak that occurred in 2013 in a low-income community in Rio de Janeiro was investigated for the presence of enteric viruses, including species A rotavirus (RVA), norovirus (NoV), astrovirus (HAstV), bocavirus (HBoV), aichivirus (AiV), and adenovirus (HAdV). Five of nine stool samples (83%) from patients were positive for HAdV, and no other enteric viruses were detected. Polymerase chain reaction products were sequenced and subjected to phylogenetic analysis, which revealed four strains and one strain of non-enteric HAdV-A12 and HAdV-F41, respectively. The HAdV-A12 nucleotide sequences shared 100% nucleotide similarity. Viral load was assessed using a TaqMan real-time PCR assay. Stool samples that were positive for HAdV-A12 had high viral loads (mean 1.9 X 107 DNA copies/g stool). All four patients with HAdV-A12 were < 25 months of age and had symptoms of fever and diarrhoea. Evaluation of enteric virus outbreaks allows the characterisation of novel or unique diarrhoea-associated viruses in regions where RVA vaccination is routinely performed. PMID:27223654

  19. Enteric disease in broiler chickens following experimental infection with chicken parvovirus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Day-old broiler chickens were inoculated orally with the chicken parvovirus strain, chicken parvovirus-P1. In four independent experiments, characteristic clinical signs of enteric disease including watery, mustard color diarrhea and growth retardation were observed following infection. The virus wa...

  20. Virus separation using membranes.

    PubMed

    Grein, Tanja A; Michalsky, Ronald; Czermak, Peter

    2014-01-01

    Industrial manufacturing of cell culture-derived viruses or virus-like particles for gene therapy or vaccine production are complex multistep processes. In addition to the bioreactor, such processes require a multitude of downstream unit operations for product separation, concentration, or purification. Similarly, before a biopharmaceutical product can enter the market, removal or inactivation of potential viral contamination has to be demonstrated. Given the complexity of biological solutions and the high standards on composition and purity of biopharmaceuticals, downstream processing is the bottleneck in many biotechnological production trains. Membrane-based filtration can be an economically attractive and efficient technology for virus separation. Viral clearance, for instance, of up to seven orders of magnitude has been reported for state of the art polymeric membranes under best conditions.This chapter summarizes the fundamentals of virus ultrafiltration, diafiltration, or purification with adsorptive membranes. In lieu of an impractical universally applicable protocol for virus filtration, application of these principles is demonstrated with two examples. The chapter provides detailed methods for production, concentration, purification, and removal of a rod-shaped baculovirus (Autographa californica M nucleopolyhedrovirus, about 40 × 300 nm in size, a potential vector for gene therapy, and an industrially important protein expression system) or a spherical parvovirus (minute virus of mice, 22-26 nm in size, a model virus for virus clearance validation studies).

  1. Nuclear entry of DNA viruses

    PubMed Central

    Fay, Nikta; Panté, Nelly

    2015-01-01

    DNA viruses undertake their replication within the cell nucleus, and therefore they must first deliver their genome into the nucleus of their host cells. Thus, trafficking across the nuclear envelope is at the basis of DNA virus infections. Nuclear transport of molecules with diameters up to 39 nm is a tightly regulated process that occurs through the nuclear pore complex (NPC). Due to the enormous diversity of virus size and structure, each virus has developed its own strategy for entering the nucleus of their host cells, with no two strategies alike. For example, baculoviruses target their DNA-containing capsid to the NPC and subsequently enter the nucleus intact, while the hepatitis B virus capsid crosses the NPC but disassembles at the nuclear side of the NPC. For other viruses such as herpes simplex virus and adenovirus, although both dock at the NPC, they have each developed a distinct mechanism for the subsequent delivery of their genome into the nucleus. Remarkably, other DNA viruses, such as parvoviruses and human papillomaviruses, access the nucleus through an NPC-independent mechanism. This review discusses our current understanding of the mechanisms used by DNA viruses to deliver their genome into the nucleus, and further presents the experimental evidence for such mechanisms. PMID:26029198

  2. [Secondary aorto-enteric fistula].

    PubMed

    Giordanengo, F; Boneschi, M; Miani, S; Erba, M; Beretta, L

    1998-01-01

    Aortic graft fistula is a rare and life-threatening complication after aortic reconstruction. The incidence ranges from 0.5 to 4%, and even if the diagnosis and treatment is appropriate, the results of surgery are poor: mortality rate ranges from 14 to 70%. The optimal method of treatment is still controversial; prosthetic removal and extra-anatomic bypass has been advocated as the standard method, but more recently, because the high mortality rate associated with this procedure, some have prompted to recommend in situ aortic graft replacement as a more successful treatment. Personal experience with incidence (0.7%) outcome and mortality (57%) in 7 patients treated over a period of 6 years (1990-1996) is reported. Results from this group are compared with another group (6 patients) previously treated (1975-1982) for the same pathology. Our results after 10 years, show the same incidence (0.7 vs 0.6%) and an elevated and unchanged mortality (57 vs 66%). Better results in the management of aorto-enteric fistulas could be achieved with the removal of infected infrarenal aortic prosthetic grafts and in situ homografts replacement.

  3. Entering China: an unconventional approach.

    PubMed

    Vanhonacker, W

    1997-01-01

    Conventional wisdom has it that the best way to do business in China is through an equity joint venture (EJV) with a well-connected Chinese partner. But pioneering companies are starting a trend toward a new way to enter that market: as a wholly foreign-owned enterprise, or WFOE. Increasingly, says the author, joint ventures do not offer foreign companies what they need to succeed in China. For example, many companies want to do business nationally, but the prospects for finding a Chinese partner with national scope are poor. Moreover, there are often conflicting perceptions between partners about how to operate an EJV: Chinese companies, for example, typically have a more immediate interest in profits than foreign investors do. By contrast, the author asserts, WFOEs are faster to set up and easier to manage; and they allow managers to expand operations more rapidly. That makes them the perfect solution, right? The answer is a qualified yes. First, foreign companies will still need sources of guanxi, or social and political connections. Second, managers must take steps to avoid trampling on China's cultural or economic sovereignty. Third and perhaps most important, foreign companies must be prepared to bring something of value to China-usually in the form of jobs or new technology that can help the country develop. Companies willing to make the effort, says the author, can reap the rewards of China's burgeoning marketplace.

  4. Vaccines against enteric infections for the developing world

    PubMed Central

    Czerkinsky, Cecil; Holmgren, Jan

    2015-01-01

    Since the first licensure of the Sabin oral polio vaccine more than 50 years ago, only eight enteric vaccines have been licensed for four disease indications, and all are given orally. While mucosal vaccines offer programmatically attractive tools for facilitating vaccine deployment, their development remains hampered by several factors: —limited knowledge regarding the properties of the gut immune system during early life;—lack of mucosal adjuvants, limiting mucosal vaccine development to live-attenuated or killed whole virus and bacterial vaccines;—lack of correlates/surrogates of mucosal immune protection; and—limited knowledge of the factors contributing to oral vaccine underperformance in children from developing countries.There are now reasons to believe that the development of safe and effective mucosal adjuvants and of programmatically sound intervention strategies could enhance the efficacy of current and next-generation enteric vaccines, especially in lesser developed countries which are often co-endemic for enteric infections and malnutrition. These vaccines must be safe and affordable for the world's poorest, confer long-term protection and herd immunity, and must be able to contain epidemics. PMID:25964464

  5. Vaccines against enteric infections for the developing world.

    PubMed

    Czerkinsky, Cecil; Holmgren, Jan

    2015-06-19

    Since the first licensure of the Sabin oral polio vaccine more than 50 years ago, only eight enteric vaccines have been licensed for four disease indications, and all are given orally. While mucosal vaccines offer programmatically attractive tools for facilitating vaccine deployment, their development remains hampered by several factors: -limited knowledge regarding the properties of the gut immune system during early life; -lack of mucosal adjuvants, limiting mucosal vaccine development to live-attenuated or killed whole virus and bacterial vaccines; -lack of correlates/surrogates of mucosal immune protection; and -limited knowledge of the factors contributing to oral vaccine underperformance in children from developing countries. There are now reasons to believe that the development of safe and effective mucosal adjuvants and of programmatically sound intervention strategies could enhance the efficacy of current and next-generation enteric vaccines, especially in lesser developed countries which are often co-endemic for enteric infections and malnutrition. These vaccines must be safe and affordable for the world's poorest, confer long-term protection and herd immunity, and must be able to contain epidemics.

  6. Application and Preparation of Enteric Coating Materials

    NASA Astrophysics Data System (ADS)

    Zhou, M. M.; Wang, L.; Zhang, X. L.; Zhou, H. J.; Chen, X. Q.; Li, Y. T.; Yang, S. L.

    2017-02-01

    In this paper, polymethacrylate enteric coated materials based on the equal mass of methyl acrylic acid and ethyl acrylate as the main raw materials were synthesized through emulsion polymerization. Omeprazole Enteric-coated Capsules were prepared by the fluidized bed coating technology using above materials as enteric layer and in vitro enteric test was considered according to standard. The results showed that the material had good coverage in the surface of omeprazole isolated pellets, excellent acid resistance in artificial gastric acid environment, and reached the disintegration effect in the buffer solution of 20min. Moreover the drug release reached 88.2% and had excellent long-term storage.

  7. Environmental enteric dysfunction: An overview

    PubMed Central

    Crane, Rosie J.; Jones, Kelsey D. J.; Berkley, James A.

    2015-01-01

    Background Environmental enteric dysfunction (EED) refers to an incompletely defined syndrome of inflammation, reduced absorptive capacity, and reduced barrier function in the small intestine. It is widespread among children and adults in low- and middle-income countries. Understanding of EED and its possible consequences for health is currently limited. Objective A narrative review of the current understanding of EED: epidemiology, pathogenesis, therapies, and relevance to child health. Methods Searches for key papers and ongoing trials were conducted using PUBMED 1966–June 2014; ClinicalTrials.gov; the WHO Clinical Trials Registry; the Cochrane Library; hand searches of the references of retrieved literature; discussions with experts; and personal experience from the field. Results EED is established during infancy and is associated with poor sanitation, certain gut infections, and micronutrient deficiencies. Helicobacter pylori infection, small intestinal bacterial overgrowth (SIBO), abnormal gut microbiota, undernutrition, and toxins may all play a role. EED is usually asymptomatic, but it is important due to its association with stunting. Diagnosis is frequently by the dual sugar absorption test, although other biomarkers are emerging. EED may partly explain the reduced efficacy of oral vaccines in low- and middle-income countries and the increased risk of serious infection seen in children with undernutrition. Conclusions Despite its potentially significant impacts, it is currently unclear exactly what causes EED and how it can be treated or prevented. Ongoing trials involve nutritional supplements, water and sanitation interventions, and immunomodulators. Further research is needed to better understand this condition, which is of likely crucial importance for child health and development in low- and middle-income settings. PMID:25902619

  8. [New home enteral nutrition via jejunostomy using semi-solid enteral formula].

    PubMed

    Ogasawara, Mihoko; Makishima, Junko; Maruyama, Makishima Michio

    2014-12-01

    Home enteral nutrition via jejunostomy requires considerable time for infusion to the patients. A new method using semisolid enteral formula dramatically reduces the time of infusion. This method makes use of pectin and liquid enteral formula. The authors applied this method in two patients with total gastrectomy being given enteral nutrition at home. There were no complications such as diarrhea or abdominal pain. This new method of home enteral nutrition could enhance the patients' quality of life (QOL) by reducing the time of infusion of enteral nutrition.

  9. Novel methods for detection of foodborne viruses

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Enteric viruses such as norovirus are the number one cause of foodborne illness. Bivalve shellfish such as oysters efficiently bioconcentrate and retain theses pathogens, making raw shellfish consumption a significant risk factor for acquisition of these viruses. Recent ARS research indicates...

  10. A simple method to screen cilantro and parsley for fecal indicator viruses.

    PubMed

    Endley, Seema; Johnson, Erin; Pillai, Suresh D

    2003-08-01

    A protocol has been developed to process cilantro and parsley samples for male-specific coliphages. Coliphage recovery depended on the duration of peptone rinsing, and whether the products were intact or cut. After 60 min of rinsing with 0.1% peptone, 78% of spiked coliphages were recovered from intact cilantro samples, and 60% of the spiked coliphages were recovered from cut cilantro samples. The protocol was field tested on a limited scale using cilantro and parsley samples from six retail outlets using enrichment-based and quantitative coliphage assays. Of the 18 retail cilantro and parsley samples that were analyzed, 50% (9 of 18) of the cilantro samples were positive for male-specific coliphages using the enrichment-based assay compared to 39% (7 of 18) of the parsley samples. Using the quantitative coliphage assay, only 28% (5 of 18) of the cilantro samples were positive, and none of the parsley samples were positive. The number of male-specific coliphages ranged between 1 and 11 plaque-forming units per 10 g of cilantro samples. None of the samples was positive for Escherichia coli. The results suggest that simplified male-specific coliphage screening of herb samples is possible and that male-specific coliphages be used along with conventional bacteriological indicators to screen produce for presence of fecal contamination.

  11. Molecular Interactions in the Replication of Mouse Hepatitis Virus

    DTIC Science & Technology

    1987-05-08

    TGEV, transmissible gastroenteritis virus; CCV, canine coronavirus; FIPV, feline infectious peritonit{s virus; FECV, feline enteric coronavirus...mouse fibroblasts (Sturman and Takemoto, 1972). 8 9 Feline infectious peritonitis was initially de- scribed and shown to be transmissible in 1963...but can lead to fatal disease, basically involving either peritonitis or the central nervous system. The causative virus, feline infectious

  12. Effects of precipitation events on virus presence in groundwater

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previous studies by our group have demonstrated the presence of human enteric viruses in groundwater, and that leakage from sanitary sewers is a likely source of such contamination in urban areas. This work showed high rates of virus detection, and that virus detection was positively correlated with...

  13. Murine Cytomegalovirus Exploits Olfaction To Enter New Hosts

    PubMed Central

    Farrell, Helen E.; Lawler, Clara; Tan, Cindy S. E.; MacDonald, Kate; Bruce, Kimberley; Mach, Michael; Davis-Poynter, Nick

    2016-01-01

    ABSTRACT   Viruses transmit via the environmental and social interactions of their hosts. Herpesviruses have colonized mammals since their earliest origins, suggesting that they exploit ancient, common pathways. Cytomegaloviruses (CMVs) are assumed to enter new hosts orally, but no site has been identified. We show by live imaging that murine CMV (MCMV) infects nasally rather than orally, both after experimental virus uptake and during natural transmission. Replication-deficient virions revealed the primary target as olfactory neurons. Local, nasal replication by wild-type MCMV was not extensive, but there was rapid systemic spread, associated with macrophage infection. A long-term, transmissible infection was then maintained in the salivary glands. The viral m131/m129 chemokine homolog, which influences tropism, promoted salivary gland colonization after nasal entry but was not required for entry per se. The capacity of MCMV to transmit via olfaction, together with previous demonstrations of experimental olfactory infection by murid herpesvirus 4 (MuHV-4) and herpes simplex virus 1 (HSV-1), suggest that this is a common, conserved route of mammalian herpesvirus entry. PMID:27118588

  14. Virus Infections in the Nervous System

    PubMed Central

    Koyuncu, Orkide O.; Hogue, Ian B.; Enquist, Lynn W.

    2013-01-01

    Virus infections usually begin in peripheral tissues and can invade the mammalian nervous system (NS), spreading into the peripheral (PNS) and more rarely the central nervous systems (CNS). The CNS is protected from most virus infections by effective immune responses and multi-layer barriers. However, some viruses enter the NS with high efficiency via the bloodstream or by directly infecting nerves that innervate peripheral tissues, resulting in debilitating direct and immune-mediated pathology. Most viruses in the NS are opportunistic or accidental pathogens, but a few, most notably the alpha herpesviruses and rabies virus, have evolved to enter the NS efficiently and exploit neuronal cell biology. Remarkably, the alpha herpesviruses can establish quiescent infections in the PNS, with rare but often fatal CNS pathology. Here we review how viruses gain access to and spread in the well-protected CNS, with particular emphasis on alpha herpesviruses, which establish and maintain persistent NS infections. PMID:23601101

  15. Vaccination against enteric septicaemia of catfish

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Edwardsiella ictaluri, the causative agent of enteric septicemia (ESC) of catfish, is one of the most economically important diseases of cultured channel catfish. In 2002, Wagner and coworkers reported that enteric septicemia of catfish (ESC) and columnaris (Flavobacterium columnaris) were the two m...

  16. Enteral nutrition after bone marrow transplantation

    PubMed Central

    Papadopoulou, A; MacDonald, A; Williams, M; Darbyshire, P; Booth, I

    1997-01-01

    Accepted 16 April 1997
 Nutritional insult after bone marrow transplantation (BMT) is complex and its nutritional management challenging. Enteral nutrition is cheaper and easier to provide than parenteral nutrition, but its tolerance and effectiveness in reversing nutritional depletion after BMT is poorly defined. Nutritional status, wellbeing, and nutritional biochemistry were prospectively assessed in 21 children (mean age 7.5 years; 14 boys) who received nasogastric feeding after BMT (mean duration 17 days) and in eight children (mean age 8 years, four boys) who refused enteral nutrition and who received dietetic advice only.
 Enteral nutrition was stopped prematurely in eight patients. Greater changes in weight and mid upper arm circumference were observed in the enteral nutrition group, while positive correlations were found between the duration of feeds and increase in weight and in mid upper arm circumference. Vomiting and diarrhoea had a similar incidence in the two groups, while fever and positive blood cultures occurred more frequently in the dietetic advice group. Diarrhoea occurring during enteral nutrition was not associated with fat malabsorption, while carbohydrate malabsorption was associated with rotavirus infection only. Enteral feeding did not, however, affect bone marrow recovery, hospital stay, general wellbeing, or serum albumin concentrations. Hypomagnesaemia, hypophosphataemia, zinc and selenium deficiency were common in both groups. In conclusion, enteral nutrition, when tolerated, is effective in limiting nutritional insult after BMT. With existing regimens nutritional biochemistry should be closely monitored in order to provide supplements when required.

 PMID:9301351

  17. Intestinal endocrine cells in radiation enteritis

    SciTech Connect

    Pietroletti, R.; Blaauwgeers, J.L.; Taat, C.W.; Simi, M.; Brummelkamp, W.H.; Becker, A.E. )

    1989-08-01

    In this study, the intestinal endocrine cells were investigated in 13 surgical specimens affected by radiation enteritis. Endocrine cells were studied by means of Grimelius' silver staining and immunostaining for chromogranin, a general marker of endocrine cells. Positively stained cells were quantified by counting their number per unit length of muscularis mucosa. Results in radiation enteritis were compared with matched control specimens by using Student's t test. Chromogranin immunostaining showed a statistically significant increase of endocrine cells in radiation enteritis specimens compared with controls both in small and large intestine (ileum, 67.5 +/- 23.5 cells per unit length of muscularis mucosa in radiation enteritis versus 17.0 +/- 6.1 in controls; colon, 40.9 +/- 13.7 cells per unit length of muscularis mucosa in radiation enteritis versus 9.5 +/- 4.1 in controls--p less than 0.005 in both instances). Increase of endocrine cells was demonstrated also by Grimelius' staining; however, without reaching statistical significance. It is not clear whether or not the increase of endocrine cells in radiation enteritis reported in this study is caused by a hyperplastic response or by a sparing phenomenon. We should consider that increased endocrine cells, when abnormally secreting their products, may be involved in some of the clinical features of radiation enteropathy. In addition, as intestinal endocrine cells produce trophic substances to the intestine, their increase could be responsible for the raised risk of developing carcinoma of the intestine in long standing radiation enteritis.

  18. [Enteric nervous system and Parkinson's disease].

    PubMed

    Paillusson, S; Lebouvier, T; Pouclet, H; Coron, E; Bruley des Varannes, S; Damier, P; Neunlist, M; Derkinderen, P

    2012-06-01

    It has become increasingly evident over the last years that Parkinson's disease is a multicentric neurodegenerative disease that affects several neuronal structures outside the substantia nigra, among which is the enteric nervous system. The aims of the present article are to discuss the role of the enteric nervous system lesions in pathology spreading (Braak's hypothesis) and in the gastrointestinal dysfunction encountered in Parkinson's disease. Owing to its accessibility to biopsies, we further discuss the use of the enteric nervous system as an original source of biomarker in Parkinson's disease.

  19. [Therapy of radiation enteritis--current challenges].

    PubMed

    Baranyai, Zsolt; Sinkó, Dániel; Jósa, Valéria; Zaránd, Attila; Teknos, Dániel

    2011-07-10

    Radiation enteritis is one of the most feared complications after abdominal and pelvic radiation therapy. The incidence varies from 0.5 to 5%. It is not rare that the slowly progressing condition will be fatal. During a period of 13 years 24 patients were operated due to the complication of radiation enteritis. Despite different types of surgery repeated operation was required in 25% of cases and finally 4 patients died. Analyzing these cases predisposing factors and different therapeutic options of this condition are discussed. Treatment options of radiation induced enteritis are limited; however, targeted therapy significantly improves the outcome. Cooperation between oncologist, gastroenterologist and surgeon is required to establish adequate therapeutic plan.

  20. Genomic characterization of a circovirus associated with fatal hemorrhagic enteritis in dog, Italy.

    PubMed

    Decaro, Nicola; Martella, Vito; Desario, Costantina; Lanave, Gianvito; Circella, Elena; Cavalli, Alessandra; Elia, Gabriella; Camero, Michele; Buonavoglia, Canio

    2014-01-01

    Dog circovirus (DogCV) was identified in an outbreak of enteritis in pups in Italy. The disease was observed in 6 young dachshunds pups of a litter from a breeding kennel and caused the death of 2 dogs. Upon full-genome analysis, the virus detected in one of the dead pups (strain Bari/411-13) was closely related to DogCVs that have been recently isolated in the USA. The present study, if corroborated by further reports, could represent a useful contribution to the knowledge of the pathogenic potential of DogCV and its association with enteritis in dogs.

  1. Genomic Characterization of a Circovirus Associated with Fatal Hemorrhagic Enteritis in Dog, Italy

    PubMed Central

    Decaro, Nicola; Martella, Vito; Desario, Costantina; Lanave, Gianvito; Circella, Elena; Cavalli, Alessandra; Elia, Gabriella; Camero, Michele; Buonavoglia, Canio

    2014-01-01

    Dog circovirus (DogCV) was identified in an outbreak of enteritis in pups in Italy. The disease was observed in 6 young dachshunds pups of a litter from a breeding kennel and caused the death of 2 dogs. Upon full-genome analysis, the virus detected in one of the dead pups (strain Bari/411–13) was closely related to DogCVs that have been recently isolated in the USA. The present study, if corroborated by further reports, could represent a useful contribution to the knowledge of the pathogenic potential of DogCV and its association with enteritis in dogs. PMID:25147946

  2. REVERSETRANSCIPTION-PCR ASSAYS FOR THE DETECTION OF BOVINE ENTERIC CALICIVIRUSES (BEC) AND ANALYSIS OF THE GENETIC RELATIONSHIPS AMONG BEC AND HUMAN CALICIVIRUSES

    EPA Science Inventory

    Two genetically distinct bovine enteric caliciviruses (BEC) have been identified: the Norwalk-like-viruses (NLV), which are genetically related to human NLV, and the distinct NB-like BEC, which is most closely related to Sapporo-like viruses and lagoviruses, but potentially may ...

  3. Enteral Nutrition and Acute Pancreatitis: A Review

    PubMed Central

    Spanier, B. W. M.; Bruno, M. J.; Mathus-Vliegen, E. M. H.

    2011-01-01

    Introduction. In patients with acute pancreatitis (AP), nutritional support is required if normal food cannot be tolerated within several days. Enteral nutrition is preferred over parenteral nutrition. We reviewed the literature about enteral nutrition in AP. Methods. A MEDLINE search of the English language literature between 1999–2009. Results. Nasogastric tube feeding appears to be safe and well tolerated in the majority of patients with severe AP, rendering the concept of pancreatic rest less probable. Enteral nutrition has a beneficial influence on the outcome of AP and should probably be initiated as early as possible (within 48 hours). Supplementation of enteral formulas with glutamine or prebiotics and probiotics cannot routinely be recommended. Conclusions. Nutrition therapy in patients with AP emerged from supportive adjunctive therapy to a proactive primary intervention. Large multicentre studies are needed to confirm the safety and effectiveness of nasogastric feeding and to investigate the role of early nutrition support. PMID:20811543

  4. [Indications and practice of enteral nutrition].

    PubMed

    Hallay, Judit; Nagy, Dániel; Fülesdi, Béla

    2014-12-21

    Malnutrition in hospitalised patients has a significant and disadvantageous impact on treatment outcome. If possible, enteral nutrition with an energy/protein-balanced nutrient should be preferred depending on the patient's condition, type of illness and risk factors. The aim of the nutrition therapy is to increase the efficacy of treatment and shorten the length of hospital stay in order to ensure rapid rehabilitation. In the present review the authors summarize the most important clinical and practical aspects of enteral nutrition therapy.

  5. Clinical Significance of Enteric Protozoa in the Immunosuppressed Human Population

    PubMed Central

    Stark, D.; Barratt, J. L. N.; van Hal, S.; Marriott, D.; Harkness, J.; Ellis, J. T.

    2009-01-01

    Summary: Globally, the number of immunosuppressed people increases each year, with the human immunodeficiency virus (HIV) pandemic continuing to spread unabated in many parts of the world. Immunosuppression may also occur in malnourished persons, patients undergoing chemotherapy for malignancy, and those receiving immunosuppressive therapy. Components of the immune system can be functionally or genetically abnormal as a result of acquired (e.g., caused by HIV infection, lymphoma, or high-dose steroids or other immunosuppressive medications) or congenital illnesses, with more than 120 congenital immunodeficiencies described to date that either affect humoral immunity or compromise T-cell function. All individuals affected by immunosuppression are at risk of infection by opportunistic parasites (such as the microsporidia) as well as those more commonly associated with gastrointestinal disease (such as Giardia). The outcome of infection by enteric protozoan parasites is dependent on absolute CD4+ cell counts, with lower counts being associated with more severe disease, more atypical disease, and a greater risk of disseminated disease. This review summarizes our current state of knowledge on the significance of enteric parasitic protozoa as a cause of disease in immunosuppressed persons and also provides guidance on recent advances in diagnosis and therapy for the control of these important parasites. PMID:19822892

  6. Porcine circovirus type 2 (PCV2) enteric disease: an independent condition or part of the systemic disease?

    PubMed

    Baró, J; Segalés, J; Martínez, J

    2015-03-23

    Intestinal disorders in growing and finishing pigs have been associated with several infectious agents, including Porcine circovirus type 2 (PCV2). This virus has been mainly related with PCV2-systemic disease (PCV2-SD); nevertheless, some authors have suggested a possible restricted intestinal infection of this virus associated with enteric clinical signs. This condition has been referred as PCV2-enteric disease (PCV2-ED). The present study analysed retrospectively, from a pathological point of view, the relation between intestinal disorders and PCV2 infection in nursery and growing-finishing pigs. Among the 96 selected pigs suffering from enteric disease and submitted for necropsy between 1998 and 2011, the most prevalent enteric lesions were catarrhal enteritis/colitis (77.1%), followed by fibrinous lesions (11.5%), granulomatous inflammation (4.2%) and other lesions such as haemorrhages or ulceration (4.2%). Seventy-two pigs (75%) were positive for PCV2 by in situ hybridization (ISH). Among positive pigs for PCV2 ISH, 39 animals suffered from PCV2-SD and 33 had no lymphoid lesions but low amount of viral nucleic acid in several lymphoid tissues, therefore, these animals did not qualify for PCVD-ED. In conclusion, all animals with enteric disorders that were positive to PCV2 by ISH had evidence of viral systemic infection. These results suggest that PCV2-ED is probably a negligible condition and PCV2 mainly contributes to enteric clinical disorders in relation to PCV2-SD occurrence.

  7. Transport of viruses through saturated and unsaturated columns packed with sand

    USGS Publications Warehouse

    Anders, R.; Chrysikopoulos, C.V.

    2009-01-01

    Laboratory-scale virus transport experiments were conducted in columns packed with sand under saturated and unsaturated conditions. The viruses employed were the male-specific RNA coliphage, MS2, and the Salmonella typhimurium phage, PRD1. The mathematical model developed by Sim and Chrysikopoulos (Water Resour Res 36:173-179, 2000) that accounts for processes responsible for removal of viruses during vertical transport in one-dimensional, unsaturated porous media was used to fit the data collected from the laboratory experiments. The liquid to liquid-solid and liquid to air-liquid interface mass transfer rate coefficients were shown to increase for both bacteriophage as saturation levels were reduced. The experimental results indicate that even for unfavorable attachment conditions within a sand column (e.g., phosphate-buffered saline solution; pH = 7.5; ionic strength = 2 mM), saturation levels can affect virus transport through porous media. ?? Springer Science+Business Media B.V. 2008.

  8. Hypophosphatemia associated with enteral alimentation in cats.

    PubMed

    Justin, R B; Hohenhaus, A E

    1995-01-01

    Hypophosphatemia is uncommon in cats, but it has been reported in association with diabetes mellitus and hepatic lipidosis, where it can cause hemolysis, rhabdomyopathy, depression, seizures, and coma. The purpose of this article is to describe 9 cats that developed low serum phosphorus concentrations (< 2.5 mg/dL) subsequent to enteral alimentation. Serum biochemical analyses from more than 6,000 cats were reviewed. The medical records of all cats with hypophosphatemia were examined for history of enteral alimentation; diabetic cats were excluded from the study. Nine cats, ranging in age from 3 to 17 years, were identified. All cats had normal serum phosphorus concentrations before tube feeding began. Onset of hypophosphatemia occurred 12 to 72 hours after initiation of enteral alimentation, and the nadir for phosphorus concentrations ranged from 0.4 to 2.4 mg/dL. Hemolysis occurred in 6 of the 9 cats. Hypophosphatemia secondary to enteral alimentation is an uncommon clinical finding in cats. Cats with high alanine aminotransferase activity, hyperbilirubinemia, and weight loss should be closely monitored for hypophosphatemia during the first 72 hours of enteral alimentation.

  9. Qualitative release assessment to estimate the likelihood of henipavirus entering the United Kingdom.

    PubMed

    Snary, Emma L; Ramnial, Vick; Breed, Andrew C; Stephenson, Ben; Field, Hume E; Fooks, Anthony R

    2012-01-01

    The genus Henipavirus includes Hendra virus (HeV) and Nipah virus (NiV), for which fruit bats (particularly those of the genus Pteropus) are considered to be the wildlife reservoir. The recognition of henipaviruses occurring across a wider geographic and host range suggests the possibility of the virus entering the United Kingdom (UK). To estimate the likelihood of henipaviruses entering the UK, a qualitative release assessment was undertaken. To facilitate the release assessment, the world was divided into four zones according to location of outbreaks of henipaviruses, isolation of henipaviruses, proximity to other countries where incidents of henipaviruses have occurred and the distribution of Pteropus spp. fruit bats. From this release assessment, the key findings are that the importation of fruit from Zone 1 and 2 and bat bushmeat from Zone 1 each have a Low annual probability of release of henipaviruses into the UK. Similarly, the importation of bat meat from Zone 2, horses and companion animals from Zone 1 and people travelling from Zone 1 and entering the UK was estimated to pose a Very Low probability of release. The annual probability of release for all other release routes was assessed to be Negligible. It is recommended that the release assessment be periodically re-assessed to reflect changes in knowledge and circumstances over time.

  10. Qualitative Release Assessment to Estimate the Likelihood of Henipavirus Entering the United Kingdom

    PubMed Central

    Snary, Emma L.; Ramnial, Vick; Breed, Andrew C.; Stephenson, Ben; Field, Hume E.; Fooks, Anthony R.

    2012-01-01

    The genus Henipavirus includes Hendra virus (HeV) and Nipah virus (NiV), for which fruit bats (particularly those of the genus Pteropus) are considered to be the wildlife reservoir. The recognition of henipaviruses occurring across a wider geographic and host range suggests the possibility of the virus entering the United Kingdom (UK). To estimate the likelihood of henipaviruses entering the UK, a qualitative release assessment was undertaken. To facilitate the release assessment, the world was divided into four zones according to location of outbreaks of henipaviruses, isolation of henipaviruses, proximity to other countries where incidents of henipaviruses have occurred and the distribution of Pteropus spp. fruit bats. From this release assessment, the key findings are that the importation of fruit from Zone 1 and 2 and bat bushmeat from Zone 1 each have a Low annual probability of release of henipaviruses into the UK. Similarly, the importation of bat meat from Zone 2, horses and companion animals from Zone 1 and people travelling from Zone 1 and entering the UK was estimated to pose a Very Low probability of release. The annual probability of release for all other release routes was assessed to be Negligible. It is recommended that the release assessment be periodically re-assessed to reflect changes in knowledge and circumstances over time. PMID:22328916

  11. Novel canine bocavirus strain associated with severe enteritis in a dog litter.

    PubMed

    Bodewes, Rogier; Lapp, Stefanie; Hahn, Kerstin; Habierski, André; Förster, Christine; König, Matthias; Wohlsein, Peter; Osterhaus, Albert D M E; Baumgärtner, Wolfgang

    2014-11-07

    Bocaviruses are small non-enveloped viruses with a linear ssDNA genome, that belong to the genus Bocaparvovirus of the subfamiliy Parvovirinae. Bocavirus infections are associated with a wide spectrum of disease in humans and various mammalian species. Here we describe a fatal enteritis associated with infection with a novel strain of canine bocavirus 2 (CaBoV-2), that occurred in a litter of German wirehaired pointers. Necropsy performed on three puppies revealed an enteritis reminiscent of canine parvovirus associated enteritis, accompanied with signs of lymphocytolytic disease in bone marrow, spleen, lymph nodes and thymus. While other major causes of enteritis of young dogs, including canine parvovirus, were excluded, by random PCR in combination with next-generation sequencing, a novel CaBoV-2 strain was detected. Phylogenetic analysis of the genome of this novel canine bocavirus strain indicated that this virus was indeed most closely related to group 2 canine bocaviruses. Infection with canine bocavirus was confirmed by in situ hybridization, which revealed the presence of CaBoV-2 nucleic acid in the intestinal tract and lymphoid tissues of the dogs. In a small-scale retrospective analysis concerning the role of CaBoV-2 no additional cases were identified. The findings of this study provide novel insights into the pathogenicity of canine bocaviruses.

  12. Evaluation of an enteric coated aspirin preparation.

    PubMed

    Day, R O; Paull, P D; Champion, G D; Graham, G G

    1976-02-01

    The bioavailability and gastrointestinal site of release of enteric coated aspirin tablets ("Rhusal", G.P. Laboratories) were investigated following dosage with single tablets. A delay of one to more than eight hours was observed between dosage and the appearance of salicylate in plasma or saliva. This delay was decreased by pretreatment with metoclopramide. No aspirin or salicylate was detected in gastric aspirates. The mean urinary recovery of salicylate was equivalent to 92% of the administered dose. All these tests were consistent with the designed function of the enteric coating. The time course of concentrations of salicylate in saliva rather than in plasma was confirmed as a useful technique for the evaluation of different formulations of aspirin. The acceleration of gastric emptying by metoclopramide is a useful technique for the evaluation of enteric coated tablets.

  13. Isolation of avian rotaviruses from a normal turkey flock and from a flock with signs of enteric disease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    It is often difficult to obtain enteric-origin viruses as pure isolates, complicating characterization and pathogenesis investigations. Two turkey-origin rotaviruses detected via NSP4-specific RT-PCR in pooled intestinal contents were isolated by serial passage in the African green monkey kidney cel...

  14. [Artificial nutrition in children (I): enteral access].

    PubMed

    Estevão-Costa, José

    2014-01-01

    Appropriate nutritional support is crucial in the therapeutic approach of multiple conditions, which justifies the frequent and increasing use of specific access routes for enteral and parenteral nutrition. This article reviews the relevant literature concerning indications, procedures, effectiveness and complications of enteral access routes in children. The decision between gastric and postpyloric access, and between nasogastric tube and gastrostomy is thoroughly discussed. Percutaneous endoscopic gastrostomy is the most commonly used technique when a long-term gastric access is required, given its efficacy and safety although the associated morbidity is not negligible;laparoscopic gastrostomy emerges as a potentially advantageous alternative.

  15. Gastritis, Enteritis, and Colitis in Horses.

    PubMed

    Uzal, Francisco A; Diab, Santiago S

    2015-08-01

    The gastrointestinal system of horses is affected by a large variety of inflammatory infectious and noninfectious conditions. The most prevalent form of gastritis is associated with ulceration of the pars esophagea. Although the diagnostic techniques for alimentary diseases of horses have improved significantly over the past few years, difficulties still exist in establishing the causes of a significant number of enteric diseases in this species. This problem is compounded by several agents of enteric disease also being found in the intestine of clinically normal horses, which questions the validity of the mere detection of these agents in the intestine.

  16. New insights into environmental enteric dysfunction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Environmental enteric dysfunction (EED) has been recognised as an important contributing factor to physical and cognitive stunting, poor response to oral vaccines, limited resilience to acute infections and ultimately global childhood mortality. The aetiology of EED remains poorly defined but the ep...

  17. Fetal Cyst Reveling Retroperitoneal Enteric Duplication

    PubMed Central

    Ayadi, Imene Dahmane; Bezzine, Ahlem; Hamida, Emira Ben; Marrakchi, Zahra

    2017-01-01

    Retroperitoneum is a very uncommon site of enteric duplication (ED). We report a new case of retroperitoneal ED cyst suspected in utero. Prenatal ultrasound showed an abdominal cystic mass. Noncommunicating retroperitoneal ED cyst measuring 70 mm × 30 mm was resected. Histopathologic examination confirmed the diagnosis. PMID:28082784

  18. Astronaut John Glenn Enters Friendship 7

    NASA Technical Reports Server (NTRS)

    1962-01-01

    Astronaut John Glenn enters the Mercury spacecraft, Friendship 7, prior to the launch of MA-6 on February 20, 1961 and became the first American who orbited the Earth. The MA-6 mission was the first manned orbital flight boosted by the Mercury-Atlas vehicle, a modified Atlas ICBM (Intercontinental Ballistic Missile), lasted for five hours, and orbited the Earth three times.

  19. Enteric bacterial growth rates in river water.

    PubMed

    Hendricks, C W

    1972-08-01

    Enteric bacteria, including stocked strains of pathogenic species and organisms naturally present in the stream, were capable of growth in a chemostat with autoclaved river water taken 750 m below a sewage outfall. Maximal specific growth rates for all organisms occurred at 30 C, whereas culture generation times ranged between 33.3 and 116 hr. Of the six laboratory strains of enteric species used, Escherichia coli and Enterobacter aerogenes grew at generation times of 34.5 and 33.3 hr, respectively, while the remaining Proteus, Arizona, Salmonella, and Shigella spp. reproduced at a rate two to three times slower than the coliforms. Little or no growth occurred in the water at incubation temperatures of 20 and 5 C, and death was observed for Salmonella senftenberg at 20 and 5 C and for E. aerogenes and Proteus rettgeri at 5 C. When enteric bacteria naturally present in the river water were employed in similar experiments, coliform bacteria demonstrated a generation time of approximately 116 hr, whereas fecal coliforms failed to grow. Growth of the bacteria from the river demonstrated a periodicity of approximately 100 hr, which suggests that much of the growth of these organisms in the chemostat may be on the glass surfaces. This phenomenon, however, was not observed with any of the stocked enteric species. Neither the stock cultures nor the aquatic strains were capable of growth in autoclaved river water taken above the sewage outfall at the three temperatures tested.

  20. Entering a Crack: An Encounter with Gossip

    ERIC Educational Resources Information Center

    Henderson, Linda

    2014-01-01

    In this paper, I enter a crack to think otherwise about the concept "gossip". Drawing on previous scholarship engaging with Deleuzian concepts to inform research methodologies, this paper builds on this body of work. Following Deleuze and Guattari, the paper undertakes a mapping of gossip, subsequent to an encounter with a crack.…

  1. [Enteral alimentation at home: why PEG now?].

    PubMed

    Suzuki, Y; Hanyu, N; Kashiwagi, H; Kubo, T; Aoki, T

    1996-12-01

    The history of percutaneous endoscopic gastrostomy (PEG) is relatively short. In 1980, a report entitled "Gastrostomy without laparotomy: A percutaneous endoscopic technique" by Ponsky and Gaudere was first published in the Journal of Pediatric Surgery. Thereafter, PEG soon saw widespread use in Western countries because of its clinical efficacy and economy. It has been performed in about 170,000 cases annually in the US. In contrast, its spread in Japan has been extremely slow: only about 10,000 cases have undergone this procedure annually, and this number accounted for less than 5% of patients receiving enteral alimentation. The reason why PEG has not spread may be the medical insurance system in Japan and the local distaste for operation scarring. However, in consideration of the unprecedented ageing of society that is surely coming in the near future, the role of PEG in Japan must be reexamined. In this report, we presented the methodology of enteral alimentation at home by means of PEG, giving special consideration to: (1) "What points are improved by using enteral alimentation at home by means of PEG in various diseases; (2) dysphagia due to cerebral angiopathy; (3) terminal cancer; (4) otolaryngological diseases; and (5) Crohn disease. We also discussed "Why PEG is important now?" in performing enteral alimentation at home.

  2. [Enteral nutrition in the multiple trauma patient].

    PubMed

    Sánchez-Izquierdo Riera, J A; Montejo González, J C

    1992-01-01

    The hypermetabolism that develops in patients with severe polytraumatism has led to the need for an aggressive metabolic-nutritional support from the start. Parenteral Nutrition is the preferred technique in many instances, due to the doubts on the effectiveness of enteral nutrition in the control of the metabolic response and to problems of gastrointestinal tolerance derived from its administration. However, the role of enteral nutrition as an important factor which limits the development of bacterial translocation and the chain of events leading to multiorganic failure appears to be more and more well-established and is an important argument for justifying the early administration of enteral nutrition in these patients. In accordance with the accumulated experience of several authors over the past few years, enteral nutrition may be administered early in polytraumatized patients. This is not only accompanied by the evidence of acceptable gastrointestinal tolerance to the diet, but also by additional advantages compared to parenteral nutrition, such as the maintenance of trophism and immunocompetence of the digestive mucosa, the reduction of septic complications and also greater nutritional effectiveness which can be evaluated by the behaviour of the seric proteins used as nutritional evolution markers. The interest of the different diet formulae which exist at present, for example diets enriched with branched-chain amino acids, diets with added fibre, peptidic diets, specific pulmonary diets or "euglycaemic diets" is evaluated in this review. All these diets may mean an increase in the effectiveness and/or tolerance of enteral nutrition in polytraumatized patients, and also contribute to the handling of specific problems such as "stress" hyperglycaemia or the withdrawal of mechanical ventilation support. The use of specific nutrients for the digestive mucosa, such as glutamine or short chain fatty acids seems to be an important factor in the reduction of

  3. Inventing Viruses.

    PubMed

    Summers, William C

    2014-11-01

    In the nineteenth century, "virus" commonly meant an agent (usually unknown) that caused disease in inoculation experiments. By the 1890s, however, some disease-causing agents were found to pass through filters that retained the common bacteria. Such an agent was called "filterable virus," the best known being the virus that caused tobacco mosaic disease. By the 1920s there were many examples of filterable viruses, but no clear understanding of their nature. However, by the 1930s, the term "filterable virus" was being abandoned in favor of simply "virus," meaning an agent other than bacteria. Visualization of viruses by the electron microscope in the late 1930s finally settled their particulate nature. This article describes the ever-changing concept of "virus" and how virologists talked about viruses. These changes reflected their invention and reinvention of the concept of a virus as it was revised in light of new knowledge, new scientific values and interests, and new hegemonic technologies.

  4. Aquaporin-4 Water Channels in Enteric Neurons

    PubMed Central

    Thi, Mia M.; Spray, David C.; Hanani, Menachem

    2009-01-01

    Aquaporin-4 is a water channel predominantly found in astrocytes in the central nervous system and is believed to play a critical role in the formation and maintenance of the blood–brain barrier and in water secretion from the brain. As enteric glial cells were found to share several similarities with astrocytes, we hypothesized that enteric glia might also contain aquaporin-4. We used immunohistochemistry to identify aquaporin-4 in the myenteric and submucosal plexuses of the mouse and the rat colon. We found that sub-populations of neurons in both enteric plexuses were positively labeled for human aquaporin-4. Double staining of the enteric ganglia with antibodies to the neuronal marker neurofilament–heavy chain 100 and to aquaporin-4 showed that a minority of myenteric neurons were aquaporin-4 positive (about 12% in the mouse and 13% in the rat). In contrast, in the submucosal plexus significant numbers of neurons were positive for aquaporin-4 (about 79% in both the mouse and the rat). Double labeling for aquaporin-4 and for the glial marker glial fibrillary acidic protein verified that glial cells were not immunoreactive to aquaporin-4. We further confirmed our findings with additional aquaporin-4 antibodies and Western blot analysis. We found that, in addition to expressing aquaporin-4, the myenteric plexus and, to a greater extent, the submucosal plexus both expressed aquaporin-1. We conclude that neurons rather than glial cells contain aquaporin-4 in the colonic enteric plexuses. It is known that submucosal neurons control transport processes in the intestinal mucosa, and the high percentage of aquaporin-4-postive sub-mucosal neurons suggests that aquaporin-4 contributes to this function. PMID:17893913

  5. PERSISTANCE OF VIRUSES IN OYSTER (CRASSOSTREA VIRGINICA) HEMOCYTES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Seafood is listed as one of the top three causes of human virus infection by foodborne products. The goal of this study is to determine why human enteric viruses such as hepatitis A (HAV) and Norovirus (NV) readily persist within bivalves. Shellfish bioaccumulate water-borne pathogens and concentra...

  6. Hemocytes are sites of persistence for virus-contaminated oysters

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Like fecal bacteria, waterborne enteric viruses are readily bioconcentrated by bivalve shellfish. However while many bacteria decline rapidly when bivalves are placed in uncontaminated water, viruses tend to be retained within shellfish. In this study, we offer evidence that phagocytic blood cells...

  7. DEVELOPMENT OF MOLECULAR METHODS TO DETECT EMERGING VIRUSES

    EPA Science Inventory

    A large number of human enteric viruses are known to cause gastrointestinal illness and waterborne outbreaks. Many of these are emerging viruses that do not grow or grow poorly in cell culture and so molecular detectoin methods based on the polymerase chain reaction (PCR) are be...

  8. Dietary and enteral interventions for Crohn's disease.

    PubMed

    Yamamoto, Takayuki; Shimoyama, Takahiro; Kuriyama, Moeko

    2017-04-01

    It is now widely acknowledged that the intestinal bacterial flora together with genetic predisposing factors significantly contribute to the immunopathogenesis of inflammatory bowel disease (IBD) as reflected by mucosal immune dysregulation. Recently, there has been an increased interest in nutraceutical therapies, including probiotics, prebiotics and synbiotics. Other dietary interventions with low carbohydrate diet, omega-3 polyunsaturated fatty acids and glutamine have been attempted to downregulate the gut inflammatory response and thereby alleviate gastrointestinal symptoms. Enteral nutrition has been widely used as induction and maintenance therapies in the management of Crohn's disease (CD). In this review, a critical assessment of the results of clinical trial outcomes and meta-analyses was conducted to evaluate the efficacy of dietary and enteral interventions for CD.

  9. Clinical and nutritional implications of radiation enteritis

    SciTech Connect

    Beer, W.H.; Fan, A.; Halsted, C.H.

    1985-01-01

    The clinical and nutritional significance of radiation enteritis was assessed in eight patients with chronic diarrhea which followed curative doses of radiotherapy for pelvic malignancies. Steatorrhea, found in seven malnourished patients, was ascribed to ileal disease or previous surgery, or to bacterial contamination of the small intestine. Lactose intolerance, assessed by breath hydrogen excretion after oral lactose and by jejunal lactase levels, was found in six patients. In a subgroup of five patients, the administration of two different defined formula liquid diets by nasoduodenal infusion decreased fecal fluid and energy losses by about one-half. Compared to Vivonex-HN, the infusion of Criticare-HN was associated with greater likelihood of intestinal gas production but a three-fold greater utilization of protein. Intestinal malabsorption and malnutrition in radiation enteritis has diverse etiologies. Whereas nutritional support by liquid diet limits fecal fluid and energy losses, these diets differ significantly in clinical tolerance and biologic value.

  10. PRELIMINARY COMPARATIVE STUDY OF METHODS TO EXTRACT VIRUS FROM RAW AND PROCESSED SEWAGE SLUDGES

    EPA Science Inventory

    Two simple virus extraction techniques were compared to an EPA standard method for detection of human enteric viruses in raw sewage sludge and class A biosolids. The techniques were used to detect both indigenous and seeded virus from a plant that distributes class A material pr...

  11. CONCENTRATION AND PROCESSING OF WATERBORNE VIRUSES BY POSITIVE CHARGE 1MDS CARTRIDGE FILTERS AND ORGANIC FLOCCULATION

    EPA Science Inventory

    This chapter describes the most widely used virus adsorption-elution (VIRADEL) method for recovering human enteric viruses from water matrices (Fout et al., 1996). The method takes advantage of postively charged cartridge filters to concentrate viruses from water. The major adv...

  12. Viruses in non-disinfected drinking water from municipal wells are related to community rates of acute gastrointestinal illness

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Groundwater supplies for drinking water are frequently contaminated with low-levels of human enteric virus genomes, yet evidence for waterborne disease transmission is lacking. We related qPCR-measured enteric viruses in the tap water of 14 non-chlorinating communities in the U.S. to acute gastroint...

  13. Routes of early enteral nutrition following oesophagectomy.

    PubMed

    Elshaer, M; Gravante, G; White, J; Livingstone, J; Riaz, A; Al-Bahrani, A

    2016-09-01

    Introduction Oesophagectomy for cancer is a challenging procedure with a five-year overall survival rate of 15-20%. Early enteral nutrition following oesophagectomy is a crucial component of the postoperative recovery and carries a significant impact on the outcome. Different methods of enteral feeding were conducted in our unit. The aim of this study was to examine the efficacy and safety of nasojejunal tube (NJT), jejunostomy tube (JT) and pharyngostomy tube (PT) feeding after oesophagectomy. Methods A retrospective review was carried out of prospectively collected data on patients with oesophageal cancer who underwent an oesophagectomy between 2011 and 2014. The primary outcome was feeding tube related complications such as occlusion, dislocation and leak. The secondary outcomes were length of stay and 30-day morbidity. Results A total of 90 oesophagectomies were included in the study. A NJT was inserted in 41 patients (45.6%), a JT was used in 14 patients (15.5%) and a PT was the route for enteral nutrition in 35 patients (38.9%). In total, five patients (5.5%) developed tube related complications. There were no tube related complications in the NJT group but one JT patient (7.1%) developed tube related cellulitis (p=0.189) and four PT patients (11.4%) developed tube related haemorrhage (p=0.544), tube dislocation (p=0.544) or cellulitis (p=0.189). The median length of stay and 30-day postoperative morbidity were similar between the groups. Conclusions NJT feeding is a less invasive, feasible route for early enteral nutrition following oesophagectomy. A randomised controlled trial is recommended to verify these findings.

  14. Laboratory Investigation of Childhood Enteric Infections

    DTIC Science & Technology

    1989-01-01

    coli [13], although they are nonlactose fermenters. They are oxidase negative, facultatively anaerobic, and non -spore-forming. In distinction to...differ from Shigellae in requiring a much higher inoculum to produce disease. They are nonmotile and slow or non -lactose fermenting. They are lysine...Campylobacter than for other enterics because of autoag- glutination and non -specificity after heating. Complex serotyping schemes Laboratory Investigation of

  15. Routes of early enteral nutrition following oesophagectomy

    PubMed Central

    Gravante, G; White, J; Livingstone, J; Riaz, A; Al-Bahrani, A

    2016-01-01

    Introduction Oesophagectomy for cancer is a challenging procedure with a five-year overall survival rate of 15-20%. Early enteral nutrition following oesophagectomy is a crucial component of the postoperative recovery and carries a significant impact on the outcome. Different methods of enteral feeding were conducted in our unit. The aim of this study was to examine the efficacy and safety of nasojejunal tube (NJT), jejunostomy tube (JT) and pharyngostomy tube (PT) feeding after oesophagectomy. Methods A retrospective review was carried out of prospectively collected data on patients with oesophageal cancer who underwent an oesophagectomy between 2011 and 2014. The primary outcome was feeding tube related complications such as occlusion, dislocation and leak. The secondary outcomes were length of stay and 30-day morbidity. Results A total of 90 oesophagectomies were included in the study. A NJT was inserted in 41 patients (45.6%), a JT was used in 14 patients (15.5%) and a PT was the route for enteral nutrition in 35 patients (38.9%). In total, five patients (5.5%) developed tube related complications. There were no tube related complications in the NJT group but one JT patient (7.1%) developed tube related cellulitis (p=0.189) and four PT patients (11.4%) developed tube related haemorrhage (p=0.544), tube dislocation (p=0.544) or cellulitis (p=0.189). The median length of stay and 30-day postoperative morbidity were similar between the groups. Conclusions NJT feeding is a less invasive, feasible route for early enteral nutrition following oesophagectomy. A randomised controlled trial is recommended to verify these findings. PMID:27388543

  16. An Exposome Perspective on Environmental Enteric Dysfunction

    PubMed Central

    Mapesa, Job O.; Maxwell, Amy L.; Ryan, Elizabeth P.

    2015-01-01

    Background: Environmental exposures to chemicals have been shown to influence gastrointestinal function, yet little is known regarding whether chemical mixtures may be involved in the development of a subclinical enteric dysfunction found in infants and children born into poor hygiene and sanitation. Advances in gastrointestinal and immunotoxicology fields merit inclusion in complex discussions of environmental enteric dysfunction (EED) that severely affects children in developing countries. Objective: We aimed to highlight exposome approaches for investigating the potential influence of environmental chemical exposures on EED development, including a role for toxicant modulation of gut immune system and microbiome function. Discussion: A major focus on fecal–oral contamination in impoverished living conditions already exists for EED, and should now expand to include environmental chemicals such as pesticides and heavy metals that may be anthropogenic or dietary or from microbial sources. A comprehensive characterization of environmental chemical exposures prenatally and occurring in infants and young children will enhance our knowledge of any associated risks for EED and stunting. Conclusions: Integrating EED, chemical exposure, and stunting at various ages during childhood will enhance our apparent limited view when evaluating EED. Etiology and intervention studies should evaluate the suite of environmental chemical exposures as candidates in the composite of EED biomarkers. Citation: Mapesa JO, Maxwell AL, Ryan EP. 2016. An exposome perspective on environmental enteric dysfunction. Environ Health Perspect 124:1121–1126; http://dx.doi.org/10.1289/ehp.1510459 PMID:26713888

  17. Developing a formulary for enteral nutrition products.

    PubMed

    Hopefl, A W; Herrmann, V M

    1982-09-01

    A formulary for enteral nutrition products was developed at a university hospital. Advice was solicited from clinical dietetics and the medical staff. Reported important product variables were osmolality, caloric density, protein content and source, fat content and source, freedom from lactose, and, for oral supplements, available flavors. Data were also obtained from manufacturers regarding the composition of their products. Products were categorized as follows: liquid supplemental feedings, high calorie supplemental feedings, isotonic tube feedings, high caloric/high nitrogen tube feedings, high nitrogen tube feedings, and blenderized tube feedings. Bids were solicited in April 1981 (and annually thereafter) from manufacturers of the classified enteral nutrition products, and a contract was signed with the manufacturer in each category submitting the lowest bid. In contrast with previous experience, there was no loss from outdated products during the first year of the formulary. Categorizing enteral nutrition products into therapeutic categories appears to be a workable method to limit the number of products used in a hospital, thereby potentially decreasing inventory, waste, and hospital costs. The descriptive category titles also may encourage rational use of these products without promoting allegiance to a particular company or product.

  18. Enteral Nutrition in Dementia: A Systematic Review

    PubMed Central

    Brooke, Joanne; Ojo, Omorogieva

    2015-01-01

    The aim of this systematic review is to evaluate the role of enteral nutrition in dementia. The prevalence of dementia is predicted to rise worldwide partly due to an aging population. People with dementia may experience both cognitive and physical complications that impact on their nutritional intake. Malnutrition and weight loss in dementia correlates with cognitive decline and the progress of the disease. An intervention for long term eating difficulties is the provision of enteral nutrition through a Percutaneous Endoscopic Gastrostomy tube to improve both nutritional parameters and quality of life. Enteral nutrition in dementia has traditionally been discouraged, although further understanding of physical, nutritional and quality of life outcomes are required. The following electronic databases were searched: EBSCO Host, MEDLINE, PubMed, Cochrane Database of Systematic Reviews and Google Scholar for publications from 1st January 2008 and up to and including 1st January 2014. Inclusion criteria included the following outcomes: mortality, aspiration pneumonia, pressure sores, nutritional parameters and quality of life. Each study included separate analysis for patients with a diagnosis of dementia and/or neurological disease. Retrospective and prospective observational studies were included. No differences in mortality were found for patients with dementia, without dementia or other neurological disorders. Risk factors for poor survival included decreased or decreasing serum albumin levels, increasing age or over 80 years and male gender. Evidence regarding pneumonia was limited, although did not impact on mortality. No studies explored pressure sores or quality of life. PMID:25854831

  19. Development of a novel method for simultaneous concentration of viruses and protozoa from a single water sample.

    PubMed

    Haramoto, Eiji; Katayama, Hiroyuki; Asami, Mari; Akiba, Michihiro

    2012-06-01

    A novel method, electronegative membrane-vortex (EMV) method, was developed for simultaneous concentration of viruses and protozoa from a single water sample. Viruses and protozoa in a water sample were mixed with a cation solution and adsorbed on an electronegative membrane. Concentrated virus and protozoa samples were obtained as supernatant and pellet fractions, respectively, by vigorous vortex mixing of the membrane and centrifugation of the eluted material. The highest recovery efficiencies of model microbes from river water and tap water by this EMV method were obtained using a mixed cellulose ester membrane with a pore size of 0.45 μm (Millipore) as the electronegative membrane and MgCl(2) as the cation solution. The recovery was 27.7-86.5% for poliovirus, 25.7-68.3% for coliphage Qβ, 28.0-60.0% for Cryptosporidium oocysts, and 35.0-53.0% for Giardia cysts. The EMV method detected successfully indigenous viruses and protozoa in wastewater and river water samples from the Kofu basin, Japan, showing an overall positive rate of 100% (43/43) for human adenovirus, 79% (34/43) for norovirus GI, 65% (28/43) for norovirus GII, 23% (10/43) for Cryptosporidium oocysts, and 60% (26/43) for Giardia cysts. By direct DNA sequencing, a total of four genotypes (AI, AII, B, and G) of Giardia intestinalis were identified in the water samples, indicating that the river water was contaminated with feces from various mammals, including humans.

  20. Why are enteric ganglia so small? Role of differential adhesion of enteric neurons and enteric neural crest cells.

    PubMed Central

    Rollo, Benjamin N.; Zhang, Dongcheng; Simkin, Johanna E.; Menheniott, Trevelyan R.; Newgreen, Donald F.

    2015-01-01

    The avian enteric nervous system (ENS) consists of a vast number of unusually small ganglia compared to other peripheral ganglia. Each ENS ganglion at mid-gestation has a core of neurons and a shell of mesenchymal precursor/glia-like enteric neural crest (ENC) cells. To study ENS cell ganglionation we isolated midgut ENS cells by HNK-1 fluorescence-activated cell sorting (FACS) from E5 and E8 quail embryos, and from E9 chick embryos. We performed cell-cell aggregation assays which revealed a developmentally regulated functional increase in ENS cell adhesive function, requiring both Ca 2+ -dependent and independent adhesion. This was consistent with N-cadherin and NCAM labelling. Neurons sorted to the core of aggregates, surrounded by outer ENC cells, showing that neurons had higher adhesion than ENC cells. The outer surface of aggregates became relatively non-adhesive, correlating with low levels of NCAM and N-cadherin on this surface of the outer non-neuronal ENC cells. Aggregation assays showed that ENS cells FACS selected for NCAM-high and enriched for enteric neurons formed larger and more coherent aggregates than unsorted ENS cells. In contrast, ENS cells of the NCAM-low FACS fraction formed small, disorganised aggregates.  This suggests a novel mechanism for control of ENS ganglion morphogenesis where i) differential adhesion of ENS neurons and ENC cells controls the core/shell ganglionic structure and ii) the ratio of neurons to ENC cells dictates the equilibrium ganglion size by generation of an outer non-adhesive surface. PMID:26064478

  1. Performance of viruses and bacteriophages for fecal source determination in a multi-laboratory, comparative study.

    PubMed

    Harwood, Valerie J; Boehm, Alexandria B; Sassoubre, Lauren M; Vijayavel, Kannappan; Stewart, Jill R; Fong, Theng-Theng; Caprais, Marie-Paule; Converse, Reagan R; Diston, David; Ebdon, James; Fuhrman, Jed A; Gourmelon, Michele; Gentry-Shields, Jennifer; Griffith, John F; Kashian, Donna R; Noble, Rachel T; Taylor, Huw; Wicki, Melanie

    2013-11-15

    An inter-laboratory study of the accuracy of microbial source tracking (MST) methods was conducted using challenge fecal and sewage samples that were spiked into artificial freshwater and provided as unknowns (blind test samples) to the laboratories. The results of the Source Identification Protocol Project (SIPP) are presented in a series of papers that cover 41 MST methods. This contribution details the results of the virus and bacteriophage methods targeting human fecal or sewage contamination. Human viruses used as source identifiers included adenoviruses (HAdV), enteroviruses (EV), norovirus Groups I and II (NoVI and NoVII), and polyomaviruses (HPyVs). Bacteriophages were also employed, including somatic coliphages and F-specific RNA bacteriophages (FRNAPH) as general indicators of fecal contamination. Bacteriophage methods targeting human fecal sources included genotyping of FRNAPH isolates and plaque formation on bacterial hosts Enterococcus faecium MB-55, Bacteroides HB-73 and Bacteroides GB-124. The use of small sample volumes (≤50 ml) resulted in relatively insensitive theoretical limits of detection (10-50 gene copies or plaques × 50 ml(-1)) which, coupled with low virus concentrations in samples, resulted in high false-negative rates, low sensitivity, and low negative predictive values. On the other hand, the specificity of the human virus methods was generally close to 100% and positive predictive values were ∼40-70% with the exception of NoVs, which were not detected. The bacteriophage methods were generally much less specific toward human sewage than virus methods, although FRNAPH II genotyping was relatively successful, with 18% sensitivity and 85% specificity. While the specificity of the human virus methods engenders great confidence in a positive result, better concentration methods and larger sample volumes must be utilized for greater accuracy of negative results, i.e. the prediction that a human contamination source is absent.

  2. Zika Virus

    MedlinePlus

    Zika is a virus that is spread mostly by mosquitoes. A pregnant mother can pass it to ... through blood transfusions. There have been outbreaks of Zika virus in the United States, Africa, Southeast Asia, ...

  3. Chikungunya virus

    MedlinePlus

    Chikungunya virus infection; Chikungunya ... Where Chikungunya is found Before 2013, the virus was found in Africa, Asia, Europe, and the Indian and Pacific oceans. In late 2013, outbreaks occurred for the first time in the ...

  4. Chikungunya Virus

    MedlinePlus

    ... is key! Prevent Infection. Use mosquito repellent. Chikungunya Virus Distribution Chikungunya in the U.S. What's New Surveillance ... Clinical Challenge For Travelers CDC Travelers' Health Chikungunya Virus Home Prevention Transmission Symptoms & Treatment Geographic Distribution Chikungunya ...

  5. Outbreak of duck plague (duck herpesvirus enteritis) in numerous species of captive ducks and geese in temporal conjunction with enforced biosecurity (in-house keeping) due to the threat of avian influenza A virus of the subtype Asia H5N1.

    PubMed

    Kaleta, E F; Kuczka, A; Kühnhold, A; Bunzenthal, C; Bönner, B M; Hanka, K; Redmann, T; Yilmaz, A

    2007-01-01

    The continuing westward spread of avian influenza A virus of the subtype H5N1 in free-living and domestic birds forced the European Union and the German federal government to enhance all biosecurity measures including in-house keeping of all captive birds from October 20 to December 15, 2005. Movement of captive ducks and geese of many different species from a free-range system to tight enclosures and maintenance for prolonged times in such overcrowded sheds resulted in pronounced disturbance of natural behaviour, interruption of mating and breeding activities and possibly additional stress. Under these conditions the birds developed signs of severe disease and enhanced mortality twentyfour days later. A total of 17 out of 124 (14%) adult birds and 149 out of 184 year-old birds (81 %) died during the outbreak. A herpesvirus was isolated from many organs of succumbed ducks and geese that was identified as a duck plague herpesvirus by cross neutralization test using known antisera against duck plague virus. The published host range of duck plague comprises 34 species within the order Anseriformes. We report here on additional 14 species of this order that were found to be susceptible to duck plague virus. The exact source of the herpesvirus could not identified. However, low antibody titres in some ducks at day of vaccination indicate that at least some of the birds were latently infected with a duck plague herpesvirus. The remaining healthy appearing birds were subcutaneously vaccinated with a modified live duck plague vaccine (Intervet, Boxmeer, NL) that stopped losses and resulted in seroconversion in most of the vaccinated birds.

  6. Hepadna viruses

    SciTech Connect

    Robinson, W.; Koike, K.; Will, H.

    1987-01-01

    This book examines the molecular biology, disease pathogenesis, epidemiology, and clinical features of hepadna and other viruses with hepatic tropism and outlines future directions and approaches for their management. The volume's six sections provide a review of the various features, mechanisms, and functions of these viruses, ranging from hepadna virus replication and regulation of gene expression to the structure and function of hepadna-virus gene products.

  7. Molecular interactions and immune responses between maize fine streak virus and the leafhopper vector G. nigrifrons through differential expression and RNA interference

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize fine streak virus (MFSV) is an emerging virus of maize that is transmitted by an insect vector, the leafhopper called Graminella nigrifrons. Virus transmission by the leafhopper requires that the virus enter into and multiply in insect cells, tissues and organs before being transmitted to a ne...

  8. The molecular basis of herpes simplex virus latency.

    PubMed

    Nicoll, Michael P; Proença, João T; Efstathiou, Stacey

    2012-05-01

    Herpes simplex virus type 1 is a neurotropic herpesvirus that establishes latency within sensory neurones. Following primary infection, the virus replicates productively within mucosal epithelial cells and enters sensory neurones via nerve termini. The virus is then transported to neuronal cell bodies where latency can be established. Periodically, the virus can reactivate to resume its normal lytic cycle gene expression programme and result in the generation of new virus progeny that are transported axonally back to the periphery. The ability to establish lifelong latency within the host and to periodically reactivate to facilitate dissemination is central to the survival strategy of this virus. Although incompletely understood, this review will focus on the mechanisms involved in the regulation of latency that centre on the functions of the virus-encoded latency-associated transcripts (LATs), epigenetic regulation of the latent virus genome and the molecular events that precipitate reactivation.

  9. Multiscale Modeling of Virus Entry via Receptor-Mediated Endocytosis

    NASA Astrophysics Data System (ADS)

    Liu, Jin

    2012-11-01

    Virus infections are ubiquitous and remain major threats to human health worldwide. Viruses are intracellular parasites and must enter host cells to initiate infection. Receptor-mediated endocytosis is the most common entry pathway taken by viruses, the whole process is highly complex and dictated by various events, such as virus motions, membrane deformations, receptor diffusion and ligand-receptor reactions, occurring at multiple length and time scales. We develop a multiscale model for virus entry through receptor-mediated endocytosis. The binding of virus to cell surface is based on a mesoscale three dimensional stochastic adhesion model, the internalization (endocytosis) of virus and cellular membrane deformation is based on the discretization of Helfrich Hamiltonian in a curvilinear space using Monte Carlo method. The multiscale model is based on the combination of these two models. We will implement this model to study the herpes simplex virus entry into B78 cells and compare the model predictions with experimental measurements.

  10. S-IB Stage Entering Point Barrow

    NASA Technical Reports Server (NTRS)

    1968-01-01

    The Saturn 1B first stage (S-IB) enters the NASA barge Point Barrow, in March 1968. The Marshall Space Flight Center (MSFC) utilized a number of water transportation craft to transport the Saturn stages to-and-from the manufacturing facilities and test sites, as well as delivery to the Kennedy Space Center for launch. Developed by the Marshall Space Flight Center and built by the Chrysler Corporation at Michoud Assembly Facility (MAF), the S-IB utilized the eight H-1 engines and each produced 200,000 pounds of thrust, a combined thrust of 1,600,000 pounds.

  11. West Nile virus: North American experience

    USGS Publications Warehouse

    Hofmeister, Erik K.

    2011-01-01

    West Nile virus, a mosquito-vectored flavivirus of the Japanese encephalitis serogroup, was first detected in North America following an epizootic in the New York City area in 1999. In the intervening 11 years since the arrival of the virus in North America, it has crossed the contiguous USA, entered the Canadian provinces bordering the USA, and has been reported in the Caribbean islands, Mexico, Central America and, more recently, South America. West Nile virus has been reported in over 300 species of birds in the USA and has caused the deaths of thousands of birds, local population declines of some avian species, the clinical illness and deaths of thousands of domestic horses, and the clinical disease in over 30 000 Americans and the deaths of over 1000. Prior to the emergence of West Nile virus in North America, St. Louis encephalitis virus and Dengue virus were the only other known mosquito-transmitted flaviviruses in North America capable of causing human disease. This review will discuss the North American experience with mosquito-borne flavivirus prior to the arrival of West Nile virus, the entry and spread of West Nile virus in North America, effects on wild bird populations, genetic changes in the virus, and the current state of West Nile virus transmission.

  12. Methods for virus recovery in water

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Food safety is intimately connected to water sanitary quality as water is used at almost every node in the food production process. Common contaminating pathogens in water are human enteric viruses, many of which are responsible for foodborne disease outbreaks in the United States and other high-inc...

  13. Monoclonal antibody capture enzyme-linked immunosorbent assay for detection of bovine enteric coronavirus.

    PubMed Central

    Crouch, C F; Raybould, T J; Acres, S D

    1984-01-01

    Monoclonal antibodies reactive with three different viral polypeptides were evaluated singly and in combination as the capture antibody(s) in an enzyme-linked immunosorbent assay system for the detection of bovine enteric coronavirus. Similar levels of sensitivity were found for all combinations tested. A sensitive, highly specific, and reproducible assay for the detection of bovine enteric coronavirus was developed, using a mixture of two of these monoclonal antibodies reactive with antigenic components either external or internal to the virion. These monoclonal antibodies were bound indirectly to 96-well plates via rabbit anti-mouse immunoglobulin. After sample application and incubation, virus was detected by using rabbit anti-coronavirus peroxidase conjugate followed by enzyme substrate and chromagen. Fecal samples from a single herd of cows were screened for the presence of coronavirus by this assay. Five percent of clinically normal cows were found to be shedding coronavirus. Images PMID:6325490

  14. The dengue virus type 2 envelope protein fusion peptide is essential for membrane fusion

    SciTech Connect

    Huang, Claire Y.-H.; Butrapet, Siritorn; Moss, Kelly J.; Childers, Thomas; Erb, Steven M.; Calvert, Amanda E.; Silengo, Shawn J.; Kinney, Richard M.; Blair, Carol D.; Roehrig, John T.

    2010-01-20

    The flaviviral envelope (E) protein directs virus-mediated membrane fusion. To investigate membrane fusion as a requirement for virus growth, we introduced 27 unique mutations into the fusion peptide of an infectious cDNA clone of dengue 2 virus and recovered seven stable mutant viruses. The fusion efficiency of the mutants was impaired, demonstrating for the first time the requirement for specific FP AAs in optimal fusion. Mutant viruses exhibited different growth kinetics and/or genetic stabilities in different cell types and adult mosquitoes. Virus particles could be recovered following RNA transfection of cells with four lethal mutants; however, recovered viruses could not re-infect cells. These viruses could enter cells, but internalized virus appeared to be retained in endosomal compartments of infected cells, thus suggesting a fusion blockade. Mutations of the FP also resulted in reduced virus reactivity with flavivirus group-reactive antibodies, confirming earlier reports using virus-like particles.

  15. Virus receptors: implications for pathogenesis and the design of antiviral agents.

    PubMed

    Norkin, L C

    1995-04-01

    A virus initiates infection by attaching to its specific receptor on the surface of a susceptible host cell. This prepares the way for the virus to enter the cell. Consequently, the expression of the receptor on specific cells and tissues of the host is a major determinant of the route of entry of the virus into the host and of the patterns of virus spread and pathogenesis in the host. This review emphasizes the virus-receptor interactions of human immunodeficiency virus, the rhinoviruses, the herpesviruses, and the coronaviruses. These interactions are often found to be complex and dynamic, involving multiple sites or factors on both the virus and the host cell. Also, the receptor may play an important role in virus entry per se in addition to its role in virus binding. In the cases of human immunodeficiency virus and the rhinoviruses, ingenious approaches to therapeutic strategies based on inhibiting virus attachment and entry are under development and in clinical trials.

  16. The management of patients with enteric hyperoxaluria.

    PubMed

    Asplin, John R

    2016-02-01

    Enteric hyperoxaluria is a common occurrence in the setting of fat malabsorption, usually due to intestinal resection or intestinal bypass surgery. Enhanced intestinal absorption of dietary oxalate leads to elevated renal oxalate excretion, frequently in excess of 100 mg/d (1.14 mmol/d). Patients are at increased risk of urolithiasis and loss of kidney function from oxalate nephropathy. Fat malabsorption causes increased binding of diet calcium by free fatty acids, reducing the calcium available to precipitate diet oxalate. Delivery of unabsorbed bile salts and fatty acids to the colon increases colonic permeability, the site of oxalate hyper-absorption in enteric hyperoxaluria. The combination of soluble oxalate in the intestinal lumen and increased permeability of the colonic mucosa leads to hyperoxaluria. Dietary therapy consists of limiting oxalate and fat intake. The primary medical intervention is the use of oral oxalate binding agents such as calcium salts to reduce free intestinal oxalate levels. Bile acid sequestrants can be useful in patients with ileal resection and bile acid malabsorption. Oxalate degrading bacteria provided as probiotics are being investigated but as of yet, no definite benefit has been shown with currently available preparations. The current state of medical therapy and potential future directions will be summarized in this article.

  17. Enteric vascular endothelial response to bacterial endotoxin.

    PubMed Central

    Koshi, R.; Mathan, V. I.; David, S.; Mathan, M. M.

    1993-01-01

    The response of enteric vasculature to endotoxin was examined at the ultrastructural level using a murine model of endotoxin-induced acute diarrhoea. Morphological changes indicative of endothelial damage were evident as early as 15 minutes following endotoxin challenge. These changes, characterized by widening of intercellular spaces, increased microvillous projections and the appearance of stress fibres, preceded the leucocytic response. Endothelial damage increased with time, being associated with progressive degenerative changes in the plasma membrane, cytoplasm and organelles, ultimately leading to desquamation. These latter changes were temporally associated with margination of neutrophils and platelet adhesion to the denuded subendothelium. The venules were the primary site of these changes while the capillaries were the least affected. The arterioles were markedly constricted with minimal endothelial damage. These changes suggest that the enteric vascular endothelium may be an important target organ, and the resultant endothelial injury may have implications in host responses to endotoxin. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 PMID:8292557

  18. Mycophenolate mofetil inducing remission of lupus enteritis.

    PubMed

    Al Balushi, F; Humby, F; Mahto, A; Kelly, C; Jawad, A

    2012-04-01

    We report the case of a young woman with a background history of discoid lupus who presented with abdominal pain, vomiting and intermittent diarrhoea. Physical examination revealed tenderness in the right upper quadrant with a palpable right inguinal lymph node without any other clinical signs of active lupus. Laboratory investigations showed normal inflammatory markers, positive ANA and Anti-Ro antibodies, persistent hypocomplementemia and lymphopenia, CT showed marked bowel oedema involving the small and large bowel (halo sign) with massive ascites and moderate right-sided pleural effusion. Mantoux test, AFB and TB cultures were negative. A diagnosis of lupus enteritis was made and treatment with high-dose steroids was commenced with little improvement. Treatment with cyclophosphamide was discussed but declined by the patient. Mycophenolate mofetil was commenced and resulted in significant clinical and radiological resolution. To the best of the authors' knowledge this is the first report of the successful use of mycophenolate mofetil in inducing and maintaining remission in lupus enteritis.

  19. A Randomized Controlled Trial Assessing Infectious Disease Risks from Bathing in Fresh Recreational Waters in Relation to the Concentration of Escherichia coli, Intestinal Enterococci, Clostridium perfringens, and Somatic Coliphages

    PubMed Central

    Wiedenmann, Albrecht; Krüger, Petra; Dietz, Klaus; López-Pila, Juan M.; Szewzyk, Regine; Botzenhart, Konrad

    2006-01-01

    We performed epidemiologic studies at public freshwater bathing sites in Germany to provide a better scientific basis for the definition of recreational water quality standards. A total of 2,196 participants were recruited from the local population and randomized into bathers and non-bathers. Bathers were exposed for 10 min and had to immerse their head at least three times. Water samples for microbiological analysis were collected at 20-min intervals. Unbiased concentration–response effects with no-observed-adverse-effect levels (NOAELs) were demonstrated for three different definitions of gastroenteritis and four fecal indicator organisms. Relative risks for bathing in waters with levels above NOAELs compared with nonbathing ranged from 1.8 (95% CI, 1.2–2.6) to 4.6 (95% CI, 2.1–10.1), depending on the definition of gastroenteritis. The effect of swallowing water provided additional evidence for true dose–response relationships. Based on the NOAELs, the following guide values for water quality are suggested: 100 Escherichia coli, 25 intestinal enterococci, 10 somatic coliphages, or 10 Clostridium perfringens per 100 mL. Recreational water quality standards are intended to protect the health of those consumers who are not already immune or resistant to pathogens that may be associated with indicator organisms. In contrast to current World Health Organization recommendations, we concluded that standards should be based on rates of compliance with NOAELs rather than on attributable risks determined above NOAELs, because these risks depend mainly on the unpredictable susceptibility of the cohorts. Although in theory there is no threshold in real concentration–response relationships, we demonstrated that a NOAEL approach would be a more robust and practical solution to the complex problem of setting standards. PMID:16451859

  20. Comparison of internal process control viruses for detection of food and waterborne viruses.

    PubMed

    Blanco Fernández, María Dolores; Barrios, Melina Elizabeth; Cammarata, Robertina Viviana; Torres, Carolina; Taboga, Oscar Alberto; Mbayed, Viviana Andrea

    2017-03-29

    Enteric viruses are pathogens associated with food- and waterborne outbreaks. The recovery of viruses from food or water samples is affected by the procedures applied to detect and concentrate them. The incorporation of an internal process control virus to the analyses allows monitoring the performance of the methodology. The aim of this study was to produce a recombinant adenovirus (rAdV) and apply it together with bacteriophage PP7 as process controls. The rAdV carries a DNA construction in its genome to differentiate it from wild-type adenovirus by qPCR. The stability of both control viruses was evaluated at different pH conditions. The rAdV was stable at pH 3, 7, and 10 for 18 h. PP7 infectious particles were stable at pH 7 and showed a 2.14 log reduction at pH 10 and total decay at pH 3 after 18 h. Three virus concentration methods were evaluated: hollow-fiber tap water ultrafiltration, wastewater ultracentrifugation, and elution-PEG precipitation from lettuce. Total and infectious viruses were quantified and their recoveries were calculated. Virus recovery for rAdV and PP7 by ultrafiltration showed a wide range (2.10-84.42 and 13.54-84.62%, respectively), whereas that by ultracentrifugation was 5.05-13.71 and 6.98-13.27%, respectively. The performance of ultracentrifugation to concentrate norovirus and enteroviruses present in sewage was not significantly different to the recovery of control viruses. For detection of viruses from lettuce, genomic copies of PP7 were significantly more highly recovered than adenovirus (14.74-18.82 and 0.00-3.44%, respectively). The recovery of infectious virus particles was significantly affected during sewage ultracentrifugation and concentration from lettuce. The simultaneous use of virus controls with dissimilar characteristics and behaviors might resemble different enteric viruses.

  1. 46 CFR 111.81-3 - Cables entering boxes.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Cables entering boxes. 111.81-3 Section 111.81-3...-GENERAL REQUIREMENTS Outlet Boxes and Junction Boxes § 111.81-3 Cables entering boxes. Each cable entering a box or fitting must be protected from abrasion and must meet the following: (a) Each...

  2. Studies of Infection and dissemination of Rift Valley Fever Virus in Mosquitoes

    DTIC Science & Technology

    1989-04-19

    foregut- midgut junction; and (5) preliminary studies with regard to the mosquito cell surface receptor molecule for RVF virus. Major results and...conclusions include: (1) The patterns of midgut infection, escape of virus from the midgut , and distribution of virus after entering the hemocoel in Aedes...epithelium via cells at the foregut- midgut junction. (5) We have found evidence of specific binding of components of formalin-killed RVF virus (vaccine

  3. Mechanism of Cell Culture Adaptation of an Enteric Calicivirus, the Porcine Sapovirus Cowden Strain

    PubMed Central

    Lu, Zhongyan; Yokoyama, Masaru; Chen, Ning; Oka, Tomoichiro; Jung, Kwonil; Chang, Kyeong-Ok; Annamalai, Thavamathi

    2015-01-01

    ABSTRACT The porcine sapovirus (SaV) (PoSaV) Cowden strain is one of only a few culturable enteric caliciviruses. Compared to the wild-type (WT) PoSaV Cowden strain, tissue culture-adapted (TC) PoSaV has two conserved amino acid substitutions in the RNA-dependent RNA polymerase (RdRp) and six in the capsid protein (VP1). By using the reverse-genetics system, we identified that 4 amino acid substitutions in VP1 (residues 178, 289, 324, and 328), but not the substitutions in the RdRp region, were critical for the cell culture adaptation of the PoSaV Cowden strain. The other two substitutions in VP1 (residues 291 and 295) reduced virus replication in vitro. Three-dimensional (3D) structural analysis of VP1 showed that residue 178 was located near the dimer-dimer interface, which may affect VP1 assembly and oligomerization; residues 289, 291, 324, and 328 were located at protruding subdomain 2 (P2) of VP1, which may influence virus binding to cellular receptors; and residue 295 was located at the interface of two monomeric VP1 proteins, which may influence VP1 dimerization. Although reversion of the mutation at residue 291 or 295 from that of the TC strain to that of the WT reduced virus replication in vitro, it enhanced virus replication in vivo, and the revertants induced higher-level serum and mucosal antibody responses than those induced by the TC PoSaV Cowden strain. Our findings reveal the molecular basis for PoSaV adaptation to cell culture. These findings may provide new, critical information for the cell culture adaptation of other PoSaV strains and human SaVs or noroviruses. IMPORTANCE The tissue culture-adapted porcine sapovirus Cowden strain is one of only a few culturable enteric caliciviruses. We discovered that 4 amino acid substitutions in VP1 (residues 178, 289, 324, and 328) were critical for its adaptation to LLC-PK cells. Two substitutions in VP1 (residues 291 and 295) reduced virus replication in vitro but enhanced virus replication and induced

  4. [Norwalk virus and Noro virus].

    PubMed

    Furuta, Itaru; Yamazumi, Toshiaki; Kitahashi, Toshiaki; Yagi, Kazurou; Takemura, Tukasa

    2003-01-01

    Norwalk virus and Noro virus are members of the Caliciviridae. These viruses are morphological similarity in each other and shows small round structure. These viruses also are well known as main pathogens of acute infectious gastroenteritis. Clinical features include an incubation period of 24 of 48 hours and illness period of 18 to 72 hours with vomiting and diarrhea in most patients and high secondary attack rates. Oral transmitted infection occurs contaminated water and foods. In our country, outbreak of Noro virus-related gastroenteritis are reported sometimes in hospital and nursing home from winter to early spring seasons. This article are described to the morphlogy, physical characteristics, epidemiology, and clinical manifestation relating to Norwalk virus and Noro virus.

  5. Endocytosis of simian virus 40 into the endoplasmic reticulum

    PubMed Central

    1989-01-01

    The endocytosis of SV-40 into CV-1 cells we studied using biochemical and ultrastructural techniques. The half-time of binding of [35S]methionine-radiolabeled SV-40 to CV-1 cells was 25 min. Most of the incoming virus particles remained undegraded for several hours. Electron microscopy showed that some virus entered the endosomal/lysosomal pathway via coated vesicles, while the majority were endocytosed via small uncoated vesicles. After infection at high multiplicity, one third of total cell-associated virus was observed to enter the ER, starting 1-2 h after virus application. The viruses were present in large, tubular, smooth membrane networks generated as extentions of the ER. The results describe a novel and unique membrane transport pathway that allows endocytosed viral particles to be targeted from the plasma membrane to the ER. PMID:2556405

  6. [SHORT BOWEL SYNDROME AND NUTRITIONAL ENTERAL].

    PubMed

    Ariadel Cobo, Diana; Pereira Cunill, José Luis; Socas Macías, María; Serrano Aguayo, Pilar; Gómez Liébana, Eulalia; Morales Conde, Salvador; García Luna, Pedro Pablo

    2015-12-01

    The particularity of this case is the nutritional management that has managed to avoid the use of prolonged parenteral nutrition and possible complications by placing jejunal tube at the distal end in patients with short bowel. It is a 34-year-old colecistectomizado complicated with postoperative peritonitis and dehiscence; two years he studied with small bowel obstruction, he was made de-volvulus and was complicated with two leak at different times after the second escape took place jejunostomy side double barreled shotgun level dehiscence, presented high debits by afferent loop of the terminal jejunostomy; during admission, polyurethane probe enteral feeding was inserted by the efferent loop jejunostomy. He received jejunal tube feeding laundry in the efferent loop terminal with decreased weight gain and subsequent reconstruction of intestinal transit debit proximal jejunostomy.

  7. APOLLO 10 ASTRONAUT ENTERS LUNAR MODULE SIMULATOR

    NASA Technical Reports Server (NTRS)

    1969-01-01

    Apollo 10 lunar module pilot Eugene A. Cernan prepares to enter the lunar module simulator at the Flight Crew Training Building at the NASA Spaceport. Cernan, Apollo 10 commander Thomas P. Stafford and John W. Young, command module pilot, are to be launched May 18 on the Apollo 10 mission, a dress rehearsal for a lunar landing later this summer. Cernan and Stafford are to detach the lunar module and drop to within 10 miles of the moon's surface before rejoining Young in the command/service module. Looking on as Cernan puts on his soft helmet is Snoopy, the lovable cartoon mutt whose name will be the lunar module code name during the Apollo 10 flight. The command/service module is to bear the code name Charlie Brown.

  8. Gastric emptying of enteric-coated tablets

    SciTech Connect

    Park, H.M.; Chernish, S.M.; Rosenek, B.D.; Brunelle, R.L.; Hargrove, B.; Wellman, H.N.

    1984-03-01

    To evaluate the gastric emptying time of pharmaceutical dosage forms in a clinical setting, a relatively simple dual-radionuclide technique was developed. Placebo tablets of six different combinations of shape and size were labeled with indium-111 DTPA and enteric coated. Six volunteers participated in a single-blind and crossover study. Tablets were given in the morning of a fasting stomach with 6 oz of water containing /sup 99m/Tc pertechnetate and continuously observed with a gamma camera. A scintigraph was obtained each minute. The results suggested that the size, shape, or volume of the tablet used in this study had no significant effect in the rate of gastric emptying. The tablets emptied erratically and unpredictably, depending upon their time of arrival in the stomach in relation to the occurrence of interdigestive myoelectric contractions. The method described is a relatively simple and accurate technique to allow one to follow the gastric emptying of tablets.

  9. Environmental Enteric Dysfunction in Children: A Review

    PubMed Central

    Syed, Sana; Ali, Asad; Duggan, Christopher

    2016-01-01

    Diarrheal diseases are a major cause of childhood death in resource-poor countries, killing about 760, 000 children under age 5 each year. While deaths due to diarrhea have declined dramatically, high rates of stunting and malnutrition have persisted. Environmental Enteric Dysfunction (EED) is a subclinical condition caused by constant fecal-oral contamination with resultant intestinal inflammation and villous blunting. These histological changes were first described in the 1960’s but the clinical impact of EED is only just being recognized in the context of failure of nutritional interventions and oral vaccines in resource-poor countries. We review the existing literature regarding the underlying causes of and potential interventions for EED and poor growth in children, highlighting the epidemiology, clinical and histologic classification of the entity, as well as discussing novel biomarkers and possible therapies. Future research priorities are also discussed. PMID:26974416

  10. Entering AN ERA of Synthesis of Modeling

    NASA Astrophysics Data System (ADS)

    Guerin, Stephen

    First, I believe we're entering an era of synthesis of modeling. Over the past 20 years, we've seen the proliferation of many isolated complex systems models. I think we now need tools for researchers, policy makers and the public to share models. Sharing could happen by stacking different layers of spatial agent-based models in geographic information systems and projecting interactive visualization out onto shared surfaces. Further, we need to make model authoring tools much more accessible to the point where motivated policy makers can author on their own. With the increased ability to author and share models, I believe this will allow us to scale our research to understand and manage the many interacting systems that make up our complex world...

  11. Mechanosensitivity in the enteric nervous system

    PubMed Central

    Mazzuoli-Weber, Gemma; Schemann, Michael

    2015-01-01

    The enteric nervous system (ENS) autonomously controls gut muscle activity. Mechanosensitive enteric neurons (MEN) initiate reflex activity by responding to mechanical deformation of the gastrointestinal wall. MEN throughout the gut primarily respond to compression or stretch rather than to shear force. Some MEN are multimodal as they respond to compression and stretch. Depending on the region up to 60% of the entire ENS population responds to mechanical stress. MEN fire action potentials after mechanical stimulation of processes or soma although they are more sensitive to process deformation. There are at least two populations of MEN based on their sensitivity to different modalities of mechanical stress and on their firing pattern. (1) Rapidly, slowly and ultra-slowly adapting neurons which encode compressive forces. (2) Ultra-slowly adapting stretch-sensitive neurons encoding tensile forces. Rapid adaptation of firing is typically observed after compressive force while slow adaptation or ongoing spike discharge occurs often during tensile stress (stretch). All MEN have some common properties: they receive synaptic input, are low fidelity mechanoreceptors and are multifunctional in that some serve interneuronal others even motor functions. Consequently, MEN possess processes with mechanosensitive as well as efferent functions. This raises the intriguing hypothesis that MEN sense and control muscle activity at the same time as servo-feedback loop. The mechanosensitive channel(s) or receptor(s) expressed by the different MEN populations are unknown. Future concepts have to incorporate compressive and tensile-sensitive MEN into neural circuits that controls muscle activity. They may interact to control various forms of a particular motor pattern or regulate different motor patterns independently from each other. PMID:26528136

  12. The reversibility of virus attachment to mineral surfaces

    USGS Publications Warehouse

    Loveland, J.P.; Ryan, J.N.; Amy, G.L.; Harvey, R.W.

    1996-01-01

    Virus transport through groundwater is limited by attachment to mineral surfaces and inactivation. Current virus transport models do not consider the implications of the reversibility of virus attachment to minerals. To explore the reversibility of virus attachment to mineral surfaces, we attached PRD1, a bacteriophage considered to be a good model of enteric viruses, to quartz and ferric oxyhydroxide-coated quartz surfaces over a range of pH values in equilibrium 'static columns'. Following attachment, we detached the viruses by replacing the pore solution with solutions of equal and higher pH. The extent of virus attachment followed an attachment 'edge' that occurred at a pH value about 2.5-3.5 pH units above the pH(IEP) of the mineral surfaces. Viruses attached below this edge were irreversibly attached until the pH of the detachment solution exceeded the pH value of the attachment edge. Viruses attached above this edge were reversibly attached. Derjaguin-Landau-Verwey-Overbeek (DEVO) potential energy calculations showed that the attachment edge occurred at the pH at which the potential energy of the primary minimum was near zero, implying that the position of the primary minimum (attractive or repulsive) controlled the equilibrium distribution of the viruses. The results suggest that the reversibility of virus attachment must be considered in virus transport models for accurate predictions of virus travel time.

  13. Detection of pathogenic viruses in sewage provided early warnings of hepatitis A virus and norovirus outbreaks.

    PubMed

    Hellmér, Maria; Paxéus, Nicklas; Magnius, Lars; Enache, Lucica; Arnholm, Birgitta; Johansson, Annette; Bergström, Tomas; Norder, Heléne

    2014-11-01

    Most persons infected with enterically transmitted viruses shed large amounts of virus in feces for days or weeks, both before and after onset of symptoms. Therefore, viruses causing gastroenteritis may be detected in wastewater, even if only a few persons are infected. In this study, the presence of eight pathogenic viruses (norovirus, astrovirus, rotavirus, adenovirus, Aichi virus, parechovirus, hepatitis A virus [HAV], and hepatitis E virus) was investigated in sewage to explore whether their identification could be used as an early warning of outbreaks. Samples of the untreated sewage were collected in proportion to flow at Ryaverket, Gothenburg, Sweden. Daily samples collected during every second week between January and May 2013 were pooled and analyzed for detection of viruses by concentration through adsorption to milk proteins and PCR. The largest amount of noroviruses was detected in sewage 2 to 3 weeks before most patients were diagnosed with this infection in Gothenburg. The other viruses were detected at lower levels. HAV was detected between weeks 5 and 13, and partial sequencing of the structural VP1protein identified three different strains. Two strains were involved in an ongoing outbreak in Scandinavia and were also identified in samples from patients with acute hepatitis A in Gothenburg during spring of 2013. The third strain was unique and was not detected in any patient sample. The method used may thus be a tool to detect incipient outbreaks of these viruses and provide early warning before the causative pathogens have been recognized in health care.

  14. Comparative interactomics for virus-human protein-protein interactions: DNA viruses versus RNA viruses.

    PubMed

    Durmuş, Saliha; Ülgen, Kutlu Ö

    2017-01-01

    Viruses are obligatory intracellular pathogens and completely depend on their hosts for survival and reproduction. The strategies adopted by viruses to exploit host cell processes and to evade host immune systems during infections may differ largely with the type of the viral genetic material. An improved understanding of these viral infection mechanisms is only possible through a better understanding of the pathogen-host interactions (PHIs) that enable viruses to enter into the host cells and manipulate the cellular mechanisms to their own advantage. Experimentally-verified protein-protein interaction (PPI) data of pathogen-host systems only became available at large scale within the last decade. In this study, we comparatively analyzed the current PHI networks belonging to DNA and RNA viruses and their human host, to get insights into the infection strategies used by these viral groups. We investigated the functional properties of human proteins in the PHI networks, to observe and compare the attack strategies of DNA and RNA viruses. We observed that DNA viruses are able to attack both human cellular and metabolic processes simultaneously during infections. On the other hand, RNA viruses preferentially interact with human proteins functioning in specific cellular processes as well as in intracellular transport and localization within the cell. Observing virus-targeted human proteins, we propose heterogeneous nuclear ribonucleoproteins and transporter proteins as potential antiviral therapeutic targets. The observed common and specific infection mechanisms in terms of viral strategies to attack human proteins may provide crucial information for further design of broad and specific next-generation antiviral therapeutics.

  15. Measurement and prediction of enteric methane emission.

    PubMed

    Sejian, Veerasamy; Lal, Rattan; Lakritz, Jeffrey; Ezeji, Thaddeus

    2011-01-01

    The greenhouse gas (GHG) emissions from the agricultural sector account for about 25.5% of total global anthropogenic emission. While CO(2) receives the most attention as a factor relative to global warming, CH(4), N(2)O and chlorofluorocarbons (CFCs) also cause significant radiative forcing. With the relative global warming potential of 25 compared with CO(2), CH(4) is one of the most important GHGs. This article reviews the prediction models, estimation methodology and strategies for reducing enteric CH(4) emissions. Emission of CH(4) in ruminants differs among developed and developing countries, depending on factors like animal species, breed, pH of rumen fluid, ratio of acetate:propionate, methanogen population, composition of diet and amount of concentrate fed. Among the ruminant animals, cattle contribute the most towards the greenhouse effect through methane emission followed by sheep, goats and buffalos, respectively. The estimated CH(4) emission rate per cattle, buffaloe, sheep and goat in developed countries are 150.7, 137, 21.9 and 13.7 (g/animal/day) respectively. However, the estimated rates in developing countries are significantly lower at 95.9 and 13.7 (g/animal/day) per cattle and sheep, respectively. There exists a strong interest in developing new and improving the existing CH(4) prediction models to identify mitigation strategies for reducing the overall CH(4) emissions. A synthesis of the available literature suggests that the mechanistic models are superior to empirical models in accurately predicting the CH(4) emission from dairy farms. The latest development in prediction model is the integrated farm system model which is a process-based whole-farm simulation technique. Several techniques are used to quantify enteric CH(4) emissions starting from whole animal chambers to sulfur hexafluoride (SF6) tracer techniques. The latest technology developed to estimate CH(4) more accurately is the micrometeorological mass difference technique. Because

  16. Parvovirus enteritis in vaccinated juvenile bush dogs.

    PubMed

    Janssen, D L; Bartz, C R; Bush, M; Marchwicki, R H; Grate, S J; Montali, R J

    1982-12-01

    Parvovirus enteritis developed in 10 of 17 vaccinated juvenile bush dogs (Speothos venaticus) from 4 litters in a 5-month period. Nine dogs died. The first outbreak involved 6 of 9 bush dogs from 2 litters. Each had been vaccinated with a killed feline-origin parvovirus vaccine at 11 and 14 weeks of age. The 6 affected dogs became ill at 29 weeks of age and died. The second outbreak involved a litter of 6 bush dogs. Each had been vaccinated every 2 weeks starting at 5 weeks of age. Two were isolated from the colony at 16 weeks of age for treatment of foot sores. Three of the 4 nonisolated dogs developed parvovirus enteritis at 20 weeks of age; 2 died at 6 and 8 days, respectively, after onset of signs. The 3rd outbreak involved a litter of 2 bush dogs. Both had been vaccinated every 2 to 3 weeks, starting at 6 weeks of age. One of these dogs became ill at 17 weeks and died 13 days later. A litter of 6 maned wolves (Chrysocyon brachyurus) and a litter of 3 bush dogs were isolated from their parent colonies at 13 and 15 weeks of age, respectively. Each animal had been vaccinated weekly, beginning at 8 weeks of age, using an inactivated canine-origin parvovirus vaccine. None of the isolated animals developed the disease. Serologic testing during isolation did not reveal protective titers (greater than or equal to 1:80) against canine parvovirus in the bush dogs until they were 23 weeks old, whereas protective titers developed in the maned wolves when they were 14 to 18 weeks old. One hand-raised bush dog was vaccinated weekly, beginning at 8 weeks of age, and a protective titer developed by 21 weeks of age. It was concluded that the juvenile bush dogs went through a period during which maternal antibodies interfered with immunization, yet did not protect against the disease. When the pups were isolated from the colony during this period, then vaccinated repeatedly until protective titers developed, the disease was prevented.

  17. Properties of melt extruded enteric matrix pellets.

    PubMed

    Schilling, Sandra U; Shah, Navnit H; Waseem Malick, A; McGinity, James W

    2010-02-01

    The objective of this study was to investigate the properties of enteric matrix pellets that were prepared by hot-melt extrusion in a one-step, continuous process. Five polymers (Eudragit) L100-55, L100 and S100, Aqoat grades LF and HF) were investigated as possible matrix formers, and pellets prepared with Eudragit S100 demonstrated superior gastric protection and acceptable processibility. Extruded pellets containing Eudragit S100 and up to 40% theophylline released less than 10% drug over 2h in acid, however, the processibility and yields were compromised by the high amounts of the non-melting drug material in the formulation. Efficient plasticization of Eudragit S100 was necessary to reduce the polymer's glass transition temperature and melt viscosity. Five compounds including triethyl citrate, methylparaben, polyethylene glycol 8000, citric acid monohydrate and acetyltributyl citrate were investigated in terms of plasticization efficiency and preservation of the delayed drug release properties. The aqueous solubility of the plasticizer and its plasticization efficiency impacted the drug release rate from the matrix pellets. The use of water-soluble plasticizers resulted in a loss of gastric protection, whereas low drug release rates in acid were found for pellets containing insoluble plasticizers or no plasticizer, independent of the extent of Eudragit S100 plasticization. The release rate of theophylline in buffer pH 7.4 was faster for pellets that were prepared with efficient plasticizers. The microstructure and solid-state properties of plasticized pellets were further investigated by scanning electron microscopy and powder X-ray diffraction. Pellets prepared with efficient plasticizers (TEC, methylparaben, PEG 8000) exhibited matrices of low porosity, and the drug was homogeneously dispersed in its original polymorphic form. Pellets containing ATBC or citric acid monohydrate had to be extruded at elevated temperature and showed physical instabilities in

  18. Measurement and prediction of enteric methane emission

    NASA Astrophysics Data System (ADS)

    Sejian, Veerasamy; Lal, Rattan; Lakritz, Jeffrey; Ezeji, Thaddeus

    2011-01-01

    The greenhouse gas (GHG) emissions from the agricultural sector account for about 25.5% of total global anthropogenic emission. While CO2 receives the most attention as a factor relative to global warming, CH4, N2O and chlorofluorocarbons (CFCs) also cause significant radiative forcing. With the relative global warming potential of 25 compared with CO2, CH4 is one of the most important GHGs. This article reviews the prediction models, estimation methodology and strategies for reducing enteric CH4 emissions. Emission of CH4 in ruminants differs among developed and developing countries, depending on factors like animal species, breed, pH of rumen fluid, ratio of acetate:propionate, methanogen population, composition of diet and amount of concentrate fed. Among the ruminant animals, cattle contribute the most towards the greenhouse effect through methane emission followed by sheep, goats and buffalos, respectively. The estimated CH4 emission rate per cattle, buffaloe, sheep and goat in developed countries are 150.7, 137, 21.9 and 13.7 (g/animal/day) respectively. However, the estimated rates in developing countries are significantly lower at 95.9 and 13.7 (g/animal/day) per cattle and sheep, respectively. There exists a strong interest in developing new and improving the existing CH4 prediction models to identify mitigation strategies for reducing the overall CH4 emissions. A synthesis of the available literature suggests that the mechanistic models are superior to empirical models in accurately predicting the CH4 emission from dairy farms. The latest development in prediction model is the integrated farm system model which is a process-based whole-farm simulation technique. Several techniques are used to quantify enteric CH4 emissions starting from whole animal chambers to sulfur hexafluoride (SF6) tracer techniques. The latest technology developed to estimate CH4 more accurately is the micrometeorological mass difference technique. Because the conditions under which

  19. White paper on guidelines concerning enteric nervous system stem cell therapy for enteric neuropathies⋆

    PubMed Central

    Burns, Alan J.; Goldstein, Allan M.; Newgreen, Donald F.; Stamp, Lincon; Schäfer, Karl-Herbert; Metzger, Marco; Hotta, Ryo; Young, Heather M.; Andrews, Peter W.; Thapar, Nikhil; Belkind-Gerson, Jaime; Bondurand, Nadege; Bornstein, Joel C.; Chan, Wood Yee; Cheah, Kathryn; Gershon, Michael D.; Heuckeroth, Robert O.; Hofstra, Robert M.W.; Just, Lothar; Kapur, Raj P.; King, Sebastian K.; McCann, Conor J.; Nagy, Nandor; Ngan, Elly; Obermayr, Florian; Pachnis, Vassilis; Pasricha, Pankaj J.; Sham, Mai Har; Tam, Paul; Berghe, Pieter Vanden

    2016-01-01

    Over the last 20 years, there has been increasing focus on the development of novel stem cell based therapies for the treatment of disorders and diseases affecting the enteric nervous system (ENS) of the gastrointestinal tract (so-called enteric neuropathies). Here, the idea is that ENS progenitor/stem cells could be transplanted into the gut wall to replace the damaged or absent neurons and glia of the ENS. This White Paper sets out experts’ views on the commonly used methods and approaches to identify, isolate, purify, expand and optimize ENS stem cells, transplant them into the bowel, and assess transplant success, including restoration of gut function. We also highlight obstacles that must be overcome in order to progress from successful preclinical studies in animal models to ENS stem cell therapies in the clinic. PMID:27059883

  20. Quantitative detection of human enteric adenoviruses in river water by microfluidic digital polymerase chain reaction.

    PubMed

    Kishida, Naohiro; Noda, Naohiro; Haramoto, Eiji; Kawaharasaki, Mamoru; Akiba, Michihiro; Sekiguchi, Yuji

    2014-01-01

    We describe an assay for simple and accurate quantification of human enteric adenoviruses (EAdVs) in water samples using a recently developed quantification method named microfluidic digital polymerase chain reaction (dPCR). The assay is based on automatic distribution of reaction mixture into a large number of nanolitre-volume reaction chambers and absolute copy number quantification from the number of chambers containing amplification products on the basis of Poisson statistics. This assay allows absolute quantification of target genes without the use of standard DNA. Concentrations of EAdVs in Japanese river water samples were successfully quantified by the developed dPCR assay. The EAdVs were detected in seven of the 10 samples (1 L each), and the concentration ranged from 420 to 2,700 copies/L. The quantified values closely resemble those by most probable number (MPN)-PCR and real-time PCR when standard DNA was validated by dPCR whereas they varied substantially when the standard was not validated. Accuracy and sensitivity of the dPCR was higher than those of real-time PCR and MPN-PCR. To our knowledge, this is the first study that has successfully quantified enteric viruses in river water using dPCR. This method will contribute to better understanding of existence of viruses in water.