Enumeration of total aerobic microorganisms in foods by SimPlate Total Plate Count-Color Indicator methods and conventional culture methods: collaborative study.

PubMed

Feldsine, Philip T; Leung, Stephanie C; Lienau, Andrew H; Mui, Linda A; Townsend, David E

2003-01-01

The relative efficacy of the SimPlate Total Plate Count-Color Indicator (TPC-CI) method (SimPlate 35 degrees C) was compared with the AOAC Official Method 966.23 (AOAC 35 degrees C) for enumeration of total aerobic microorganisms in foods. The SimPlate TPC-CI method, incubated at 30 degrees C (SimPlate 30 degrees C), was also compared with the International Organization for Standardization (ISO) 4833 method (ISO 30 degrees C). Six food types were analyzed: ground black pepper, flour, nut meats, frozen hamburger patties, frozen fruits, and fresh vegetables. All foods tested were naturally contaminated. Nineteen laboratories throughout North America and Europe participated in the study. Three method comparisons were conducted. In general, there was <0.3 mean log count difference in recovery among the SimPlate methods and their corresponding reference methods. Mean log counts between the 2 reference methods were also very similar. Repeatability (Sr) and reproducibility (SR) standard deviations were similar among the 3 method comparisons. The SimPlate method (35 degrees C) and the AOAC method were comparable for enumerating total aerobic microorganisms in foods. Similarly, the SimPlate method (30 degrees C) was comparable to the ISO method when samples were prepared and incubated according to the ISO method.

Comparative analyses of viable bacterial counts in foods and seawater under microplate based liquid- and conventional agar plate cultivation: increased culturability of marine bacteria under liquid cultivation.

PubMed

Shigematsu, Toru; Ueno, Shigeaki; Tsuchida, Yasuharu; Hayashi, Mayumi; Okonogi, Hiroko; Masaki, Haruhiko; Fujii, Tomoyuki

2007-12-01

Bacterial counts under liquid cultivation using 96-well microplates were performed. The counts under liquid and under solid cultivation were equivalent in foods, although the counts under liquid cultivation exceeded those under solid cultivation in seawater, suggesting that some bacteria in seawater were viable but did not form detectable colonies. Phylogenetic analysis of bacteria obtained under liquid cultivation was also performed.

Individual based simulations of bacterial growth on agar plates

NASA Astrophysics Data System (ADS)

Ginovart, M.; López, D.; Valls, J.; Silbert, M.

2002-03-01

The individual based simulator, INDividual DIScrete SIMulations (INDISIM) has been used to study the behaviour of the growth of bacterial colonies on a finite dish. The simulations reproduce the qualitative trends of pattern formation that appear during the growth of Bacillus subtilis on an agar plate under different initial conditions of nutrient peptone concentration, the amount of agar on the plate, and the temperature. The simulations are carried out by imposing closed boundary conditions on a square lattice divided into square spatial cells. The simulator studies the temporal evolution of the bacterial population possible by setting rules of behaviour for each bacterium, such as its uptake, metabolism and reproduction, as well as rules for the medium in which the bacterial cells grow, such as concentration of nutrient particles and their diffusion. The determining factors that characterize the structure of the bacterial colony patterns in the presents simulations, are the initial concentrations of nutrient particles, that mimic the amount of peptone in the experiments, and the set of values for the microscopic diffusion parameter related, in the experiments, to the amount of the agar medium.

Technical note: enumeration of mesophilic aerobes in milk: evaluation of standard official protocols and Petrifilm aerobic count plates.

PubMed

Freitas, R; Nero, L A; Carvalho, A F

2009-07-01

Enumeration of mesophilic aerobes (MA) is the main quality and hygiene parameter for raw and pasteurized milk. High levels of these microorganisms indicate poor conditions in production, storage, and processing of milk, and also the presence of pathogens. Fifteen raw and 15 pasteurized milk samples were submitted for MA enumeration by a conventional plating method (using plate count agar) and Petrifilm Aerobic Count plates (3M, St. Paul, MN), followed by incubation according to 3 official protocols: IDF/ISO (incubation at 30 degrees C for 72 h), American Public Health Association (32 degrees C for 48 h), and Brazilian Ministry of Agriculture (36 degrees C for 48 h). The results were compared by linear regression and ANOVA. Considering the results from conventional methodology, good correlation indices and absence of significant differences between mean counts were observed, independent of type of milk sample (raw or pasteurized) and incubation conditions (IDF/ISO, American Public Health Association, or Ministry of Agriculture). Considering the results from Petrifilm Aerobic Count plates, good correlation indices and absence of significant differences were only observed for raw milk samples. The microbiota of pasteurized milk interfered negatively with the performance of Petrifilm Aerobic Count plates, probably because of the presence of microorganisms that poorly reduce the dye indicator of this system.

Correlation between the neutrophil-lymphocyte count ratio and bacterial infection in patient with human immunodeficiency virus

NASA Astrophysics Data System (ADS)

Kusnadi, D.; Liwang, M. N. I.; Katu, S.; Mubin, A. H.; Halim, R.

2018-03-01

Parameters for starting antibiotic therapy such as CRP andleukocytosis are considered non-specific. Previous studies have shown the Neutrophil-Lymphocyte Count Ratio (NLCR) can serve as the basis of bacterial infection, the level of infection, and the basis of antibiotic therapy. Compared with the Procalcitonin parameter, this NLCR is rapid, an inexpensive and requires no additional sampling. To determine the correlation between The Neutrophil-LymphocyteCount Ratio to bacterial infection in HIV patients. This study was a cross-sectional observational approach to HIV subject at Wahidin Sudirohusodo and Hasanuddin University Hospital. The subjects performed routine blood, microbiology test,and blood Procalcitonin levels tests. Then performed NLCR calculations based on routine blood results. The subjects then grouped the presence or absence of bacterial infection.In 146 study subjects, there were 78 (53.4%) with bacterial infections and 68 (46.6%) without bacterial infection as controls. Subjects with bacterial infections had higher total neutrophils (84.83) compared with non-bacterial infections. Subjects with bacterial infections had total lymphocytes with an average of 8.51 lower than non-bacterial infections. Subjects with bacterial infections had higher NLCR values with an average of 12.80. The Neutrophil-Lymphocyte Count Ratio can become a marker of bacterial infection in HIV patients.

High-Throughput Quantification of Bacterial-Cell Interactions Using Virtual Colony Counts

PubMed Central

Hoffmann, Stefanie; Walter, Steffi; Blume, Anne-Kathrin; Fuchs, Stephan; Schmidt, Christiane; Scholz, Annemarie; Gerlach, Roman G.

2018-01-01

The quantification of bacteria in cell culture infection models is of paramount importance for the characterization of host-pathogen interactions and pathogenicity factors involved. The standard to enumerate bacteria in these assays is plating of a dilution series on solid agar and counting of the resulting colony forming units (CFU). In contrast, the virtual colony count (VCC) method is a high-throughput compatible alternative with minimized manual input. Based on the recording of quantitative growth kinetics, VCC relates the time to reach a given absorbance threshold to the initial cell count using a series of calibration curves. Here, we adapted the VCC method using the model organism Salmonella enterica sv. Typhimurium (S. Typhimurium) in combination with established cell culture-based infection models. For HeLa infections, a direct side-by-side comparison showed a good correlation of VCC with CFU counting after plating. For MDCK cells and RAW macrophages we found that VCC reproduced the expected phenotypes of different S. Typhimurium mutants. Furthermore, we demonstrated the use of VCC to test the inhibition of Salmonella invasion by the probiotic E. coli strain Nissle 1917. Taken together, VCC provides a flexible, label-free, automation-compatible methodology to quantify bacteria in in vitro infection assays. PMID:29497603

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

NASA Astrophysics Data System (ADS)

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

2018-01-01

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

Optimization of high count rate event counting detector with Microchannel Plates and quad Timepix readout

NASA Astrophysics Data System (ADS)

Tremsin, A. S.; Vallerga, J. V.; McPhate, J. B.; Siegmund, O. H. W.

2015-07-01

Many high resolution event counting devices process one event at a time and cannot register simultaneous events. In this article a frame-based readout event counting detector consisting of a pair of Microchannel Plates and a quad Timepix readout is described. More than 104 simultaneous events can be detected with a spatial resolution of 55 μm, while >103 simultaneous events can be detected with <10 μm spatial resolution when event centroiding is implemented. The fast readout electronics is capable of processing >1200 frames/sec, while the global count rate of the detector can exceed 5×108 particles/s when no timing information on every particle is required. For the first generation Timepix readout, the timing resolution is limited by the Timepix clock to 10-20 ns. Optimization of the MCP gain, rear field voltage and Timepix threshold levels are crucial for the device performance and that is the main subject of this article. These devices can be very attractive for applications where the photon/electron/ion/neutron counting with high spatial and temporal resolution is required, such as energy resolved neutron imaging, Time of Flight experiments in lidar applications, experiments on photoelectron spectroscopy and many others.

Identification of Lactobacillus delbrueckii and Streptococcus thermophilus Strains Present in Artisanal Raw Cow Milk Cheese Using Real-time PCR and Classic Plate Count Methods.

PubMed

Stachelska, Milena A

2017-12-04

The aim of this paper was to detect Lactobacillus delbrueckii and Streptococcus thermophilus using real-time quantitative PCR assay in 7-day ripening cheese produced from unpasteurised milk. Real-time quantitative PCR assays were designed to identify and enumerate the chosen species of lactic acid bacteria (LAB) in ripened cheese. The results of molecular quantification and classic bacterial enumeration showed a high level of similarity proving that DNA extraction was carried out in a proper way and that genomic DNA solutions were free of PCR inhibitors. These methods revealed the presence of L. delbrueckii and S. thermophilus. The real-time PCR enabled quantification with a detection of 101-103 CFU/g of product. qPCR-standard curves were linear over seven log units down to 101 copies per reaction; efficiencies ranged from 77.9% to 93.6%. Cheese samples were analysed with plate count method and qPCR in parallel. Compared with the classic plate count method, the newly developed qPCR method provided faster and species specific identification of two dairy LAB and yielded comparable quantitative results.

A generalized plate method for estimating total aerobic microbial count.

PubMed

Ho, Kai Fai

2004-01-01

The plate method outlined in Chapter 61: Microbial Limit Tests of the U.S. Pharmacopeia (USP 61) provides very specific guidance for assessing total aerobic bioburden in pharmaceutical articles. This methodology, while comprehensive, lacks the flexibility to be useful in all situations. By studying the plate method as a special case within a more general family of assays, the effects of each parameter in the guidance can be understood. Using a mathematical model to describe the plate counting procedure, a statistical framework for making more definitive statements about total aerobic bioburden is developed. Such a framework allows the laboratory scientist to adjust the USP 61 methods to satisfy specific practical constraints. In particular, it is shown that the plate method can be conducted, albeit with stricter acceptance criteria, using a test specimen quantity that is smaller than the 10 g or 10 mL prescribed in the guidance. Finally, the interpretation of results proffered by the guidance is re-examined within this statistical framework and shown to be overly aggressive.

The effect of microchannel plate gain depression on PAPA photon counting cameras

NASA Astrophysics Data System (ADS)

Sams, Bruce J., III

1991-03-01

PAPA (precision analog photon address) cameras are photon counting imagers which employ microchannel plates (MCPs) for image intensification. They have been used extensively in astronomical speckle imaging. The PAPA camera can produce artifacts when light incident on its MCP is highly concentrated. The effect is exacerbated by adjusting the strobe detection level too low, so that the camera accepts very small MCP pulses. The artifacts can occur even at low total count rates if the image has highly a concentrated bright spot. This paper describes how to optimize PAPA camera electronics, and describes six techniques which can avoid or minimize addressing errors.

Characterisation of the spoilage bacterial microbiota in oyster gills during storage at different temperatures.

PubMed

Chen, Huibin; Liu, Zhiyu; Wang, Meiying; Chen, Shaojun; Chen, Tuanwei

2013-12-01

The spoilage bacterial community in oyster gill was investigated during storage at 4, 10 and 20 °C. Aerobic plate counts and pH values were determined. Total bacterial DNA was extracted from oyster gill and bulk cells of plate count media. The major bacterial species during fresh or different temperatures storage were determined by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The initial aerobic plate count in oyster gill reached 6.70 log CFU g(-1). PCR-DGGE fingerprinting analysis of the 16S rRNA gene V3 region revealed that most of the strains in fresh oyster gill belonged to the genera Lactococcus and Enterobacter. The major spoilage bacteria at a storage temperature of 20 °C were Leuconostoc pseudomesenteroides, an uncultured bacterium, Cytophaga fermentans, Lactococcus lactis, Pseudoalteromonas sp., Enterococcus mundtii, Clostridium difficile and an uncultured Fusobacteria; those at 10 °C were Lactococcus spp., Lactobacillus curvatus, Weissella confusa and C. difficile; those at 4 °C were Lactococcus, Weissella, Enterobacter and Aeromonas. The other minor species were L. curvatus, Pseudomonas sp. and E. mundtii. Lactococcus spp. was the most common main spoilage bacteria in oyster gill during chilled storage. PCR-DGGE revealed the complexity of the bacterial microbiota and the major bacteria species in oyster gill for fresh and storage. © 2013 Society of Chemical Industry.

[Analysis on 2011 quality control results on aerobic plate count of microbiology laboratories in China].

PubMed

Han, Haihong; Li, Ning; Li, Yepeng; Fu, Ping; Yu, Dongmin; Li Zhigang; Du, Chunming; Guo, Yunchang

2015-01-01

To test the aerobic plate count examining capability of microbiology laboratories, to ensure the accuracy and comparability of quantitative bacteria examination results, and to improve the quality of monitoring. The 4 different concentration aerobic plate count piece samples were prepared and noted as I, II, III and IV. After homogeneity and stability tests, the samples were delivered to monitoring institutions. The results of I, II, III samples were logarithmic transformed, and evaluated with Z-score method using the robust average and standard deviation. The results of IV samples were evaluated as "satisfactory" when reported as < 10 CFU/piece or as "not satisfactory" otherwise. Pearson χ2 test was used to analyze the ratio results. 309 monitoring institutions, which was 99.04% of the total number, reported their results. 271 institutions reported a satisfactory result, and the satisfactory rate was 87.70%. There was no statistical difference in satisfactory rates of I, II and III samples which were 81.52%, 88.30% and 91.40% respectively. The satisfactory rate of IV samples was 93.33%. There was no statistical difference in satisfactory rates between provincial and municipal CDC. The quality control program has provided scientific data that the aerobic plate count capability of the laboratories meets the requirements of monitoring tasks.

Short communication: bulk tank total bacterial count in dairy sheep: factors of variation and relationship with somatic cell count.

PubMed

Gonzalo, C; Carriedo, J A; Beneitez, E; Juárez, M T; De La Fuente, L F; San Primitivo, F

2006-02-01

A total of 9,353 records for bulk tank total bacterial count (TBC) were obtained over 1 yr from 315 dairy ewe flocks belonging to the Sheep Improvement Consortium (CPO) in Castilla-León (Spain). Analysis of variance showed significant effects of flock, breed, month within flock, dry therapy, milking type and installation, and logSCC on logTBC. Flock and month within flock were important variation factors as they accounted for 22.0 and 22.1% of the variance, respectively. Considerable repeatability values were obtained for both random factors. Hand milking and bucket-milking machines elicited highest logTBC (5.31), whereas parlor systems with looped milkline (5.01) elicited the lowest logTBC. The implementation of dry therapy practice (5.12) showed significantly lower logTBC than when not used (5.25). Variability in logTBC among breeds ranged from 5.24 (Awassi) to 5.07 (Churra). However, clinical outbreaks of contagious agalactia did not increase TBC significantly. A statistically significant relationship was found between logTBC and logSCC, the correlation coefficient between the variables being r = 0.23. Programs for improving milk hygiene should be implemented for both total bacterial count and somatic cell count variables at the same time.

Bacterial Cleanability of Various Types of Eating Surfaces.

ERIC Educational Resources Information Center

Ridenour, Gerald M.; Armbruster, E. H.

1953-01-01

Presents a study of the capability of commercial dishwashers to remove bacteria from various kinds of service plates. Gives an account of preliminary research on the bacterial cleanability of eating surfaces of different materials by two radiological procedures--(1) radiological count, and (2) autoradiographic measurement. Among the factors…

Photon-counting detector arrays based on microchannel array plates. [for image enhancement

NASA Technical Reports Server (NTRS)

Timothy, J. G.

1975-01-01

The recent development of the channel electron multiplier (CEM) and its miniaturization into the microchannel array plate (MCP) offers the possibility of fully combining the advantages of the photographic and photoelectric detection systems. The MCP has an image-intensifying capability and the potential of being developed to yield signal outputs superior to those of conventional photomultipliers. In particular, the MCP has a photon-counting capability with a negligible dark-count rate. Furthermore, the MCP can operate stably and efficiently at extreme-ultraviolet and soft X-ray wavelengths in a windowless configuration or can be integrated with a photo-cathode in a sealed tube for use at ultraviolet and visible wavelengths. The operation of one- and two-dimensional photon-counting detector arrays based on the MCP at extreme-ultraviolet wavelengths is described, and the design of sealed arrays for use at ultraviolet and visible wavelengths is briefly discussed.

Comparison of epifluorescent viable bacterial count methods

NASA Technical Reports Server (NTRS)

Rodgers, E. B.; Huff, T. L.

1992-01-01

Two methods, the 2-(4-Iodophenyl) 3-(4-nitrophenyl) 5-phenyltetrazolium chloride (INT) method and the direct viable count (DVC), were tested and compared for their efficiency for the determination of the viability of bacterial populations. Use of the INT method results in the formation of a dark spot within each respiring cell. The DVC method results in elongation or swelling of growing cells that are rendered incapable of cell division. Although both methods are subjective and can result in false positive results, the DVC method is best suited to analysis of waters in which the number of different types of organisms present in the same sample is assumed to be small, such as processed waters. The advantages and disadvantages of each method are discussed.

The association between bedding material and the bacterial counts of Staphylococcus aureus, Streptococcus uberis and coliform bacteria on teat skin and in teat canals in lactating dairy cattle.

PubMed

Paduch, Jan-Hendrik; Mohr, Elmar; Krömker, Volker

2013-05-01

Several mastitis-causing pathogens are able to colonize the bovine teat canal. The objective of this study was to investigate the association between the treatment of sawdust bedding with a commercial alkaline conditioner and the bacterial counts on teat skin and in the teat canal. The study used a crossover design. Ten lactating Holstein cows that were free of udder infections and mastitis were included in the study. The animals were bedded on either untreated sawdust or sawdust that had been treated with a hydrated lime-based conditioner. Once a day, fresh bedding material was added. After 3 weeks, the bedding material was removed from the cubicles, fresh bedding material was provided, and the cows were rotated between the two bedding material groups. Teat skin and teat canals were sampled using the wet and dry swab technique after weeks 1, 2, 3, 4, 5 and 6. Staphylococcus aureus, Streptococcus uberis, Escherichia coli and other coliform bacteria were detected in the resulting agar plate cultures. The treatment of the bedding material was associated with the teat skin bacterial counts of Str. uberis, Esch. coli and other coliform bacteria. An association was also found between the bedding material and the teat canal bacterial counts of coliform bacteria other than Esch. coli. For Staph. aureus, no associations with the bedding material were found. In general, the addition of a hydrated lime-based conditioner to sawdust reduces the population sizes of environmental pathogens on teat skin and in teat canals.

Short communication: Bacterial counts in recycled manure solids bedding replaced daily or deep packed in freestalls.

PubMed

Sorter, D E; Kester, H J; Hogan, J S

2014-05-01

An experiment was conducted to compare bacterial counts of mastitis pathogens in deep-packed manure solids bedding with those in manure solids bedding replaced daily from mattresses. Eighteen Holstein cows were housed in 1 pen with 18 stalls. One row of 9 stalls was equipped with mattresses topped with bedding. The back one-third of these stalls toward the alleyway was covered in 25 mm of recycled manure solids, which was removed daily for the next 6 d and replaced with bedding from the brisket board and lunge space areas of stalls. The second row of 9 stalls was bedded for 3 wk with 100 to 150 mm of deep-pack recycled manure bedding from which only fecal matter was removed daily. After 3 wk, bedding treatments were changed between rows in a switchback design. Mean total gram-negative bacterial counts did not differ between treatments throughout the experiment. Coliform and Klebsiella spp. bacterial counts were lower in daily replaced bedding compared with deep pack across the experiment and on each of d 0, 1, 2, and 6. Streptococcal counts were reduced in daily replacement stalls compared with deep-pack stalls on d 0 and greater in daily replacement stalls compared with deep-pack stalls on d 1, 2, and 6. Daily replacement of recycled manure bedding from the back one-third of the stalls appeared to be an effective approach to reducing exposure to coliforms, specifically Klebsiella, but not streptococci. However, bacterial counts in bedding from both treatments were elevated throughout the trial and resulted in considerable risk for exposure to teats and development of intramammary infections. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Gingival fluid cytokine expression and subgingival bacterial counts during pregnancy and postpartum: a case series.

PubMed

Bieri, Regina Alessandri; Adriaens, Laurence; Spörri, Stefan; Lang, Niklaus P; Persson, G Rutger

2013-01-01

The aim of this study was to assess gingival fluid (GCF) cytokine messenger RNA (mRNA) levels, subgingival bacteria, and clinical periodontal conditions during a normal pregnancy to postpartum. Subgingival bacterial samples were analyzed with the checkerboard DNA-DNA hybridization method. GCF samples were assessed with real-time PCR including five proinflammatory cytokines and secretory leukocyte protease inhibitor. Nineteen pregnant women with a mean age of 32 years (S.D. ± 4 years, range 26-42) participated in the study. Full-mouth bleeding scores (BOP) decreased from an average of 41.2% (S.D. ± 18.6%) at the 12th week of pregnancy to 26.6% (S.D. ± 14.4%) at the 4-6 weeks postpartum (p < 0.001). Between week 12 and 4-6 weeks postpartum, the mean probing pocket depth changed from 2.4 mm (S.D. ± 0.4) to 2.3 mm (S.D. ± 0.3) (p = 0.34). Higher counts of Eubacterium saburreum, Parvimonas micra, Selenomonas noxia, and Staphylococcus aureus were found at week 12 of pregnancy than at the 4-6 weeks postpartum examinations (p < 0.001). During and after pregnancy, statistically significant correlations between BOP scores and bacterial counts were observed. BOP scores and GCF levels of selected cytokines were not related to each other and no differences in GCF levels of the cytokines were observed between samples from the 12th week of pregnancy to 4-6 weeks postpartum. Decreasing postpartum counts of Porphyromonas endodontalis and Pseudomonas aeruginosa were associated with decreasing levels of Il-8 and Il-1β. BOP decreased after pregnancy without any active periodontal therapy. Associations between bacterial counts and cytokine levels varied greatly in pregnant women with gingivitis and a normal pregnancy outcome. Postpartum associations between GCF cytokines and bacterial counts were more consistent. Combined assessments of gingival fluid cytokines and subgingival bacteria may provide important information on host response.

High Speed Large Format Photon Counting Microchannel Plate Imaging Sensors

NASA Astrophysics Data System (ADS)

Siegmund, O.; Ertley, C.; Vallerga, J.; Craven, C.; Popecki, M.; O'Mahony, A.; Minot, M.

The development of a new class of microchannel plate technology, using atomic layer deposition (ALD) techniques applied to a borosilicate microcapillary array is enabling the implementation of larger, more stable detectors for Astronomy and remote sensing. Sealed tubes with MCPs with SuperGenII, bialkali, GaAs and GaN photocathodes have been developed to cover a wide range of optical/UV sensing applications. Formats of 18mm and 25mm circular, and 50mm (Planacon) and 20cm square have been constructed for uses from night time remote reconnaissance and biological single-molecule fluorescence lifetime imaging microscopy, to large area focal plane imagers for Astronomy, neutron detection and ring imaging Cherenkov detection. The large focal plane areas were previously unattainable, but the new developments in construction of ALD microchannel plates allow implementation of formats of 20cm or more. Continuing developments in ALD microchannel plates offer improved overall sealed tube lifetime and gain stability, and furthermore show reduced levels of radiation induced background. High time resolution astronomical and remote sensing applications can be addressed with microchannel plate based imaging, photon time tagging detector sealed tube schemes. Photon counting imaging readouts for these devices vary from cross strip (XS), cross delay line (XDL), to stripline anodes, and pad arrays depending on the intended application. The XS and XDL readouts have been implemented in formats from 22mm, and 50mm to 20cm. Both use MCP charge signals detected on two orthogonal layers of conductive fingers to encode event X-Y positions. XDL readout uses signal propagation delay to encode positions while XS readout uses charge cloud centroiding. Spatial resolution readout of XS detectors can be better than 20 microns FWHM, with good image linearity while using low gain (<10^6), allowing high local counting rates and longer overall tube lifetime. XS tubes with electronics can encode event

Comparison of direct-plating and broth-enrichment culture methods for detection of potential bacterial pathogens in respiratory secretions.

PubMed

Kaur, Ravinder; Wischmeyer, Jareth; Morris, Matthew; Pichichero, Michael E

2017-11-01

We compared the recovery of potential respiratory bacterial pathogens and normal flora from nasopharyngeal specimens collected from children during health and at the onset of acute otitis media (AOM) by selective direct-plating and overnight broth-enrichment. Overall, 3442 nasal wash (NW) samples collected from young children were analysed from a 10-year prospective study. NWs were cultured by (1) direct-plating to TSAII/5 % sheep blood agar and chocolate agar plates and (2) overnight broth-enrichment in BacT/ALERT SA-broth followed by plating. Standard microbiology techniques were applied to identify three dominant respiratory bacterial pathogens: Streptococcus pneumoniae (Spn), Haemophilus influenzae (Hflu) and Moraxella catarrhalis (Mcat) as well as two common nasal flora, Staphylococcus aureus (SA) and alpha-haemolytic Streptococci (AHS).Results/Key findings. Direct-plating of NW resulted in isolation of Spn from 37.8 %, Hflu from 13.6 % and Mcat from 33.2 % of samples. In comparison, overnight broth-enrichment isolated fewer Spn (30.1 %), Hflu (6.2 %) and Mcat (16.2 %) (P<0.001-0.0001). Broth-enrichment resulted in significant increased isolation of SA (6.0 %) and AHS (30.1 %) (P<0.0001). Competition between bacterial species in broth when both species were detected by direct-plating was assessed, and it was found that SA and AHS out-competed other species during broth-enrichment when samples were collected from healthy children but not during AOM. In middle ear fluids (MEF) at the onset of AOM, broth-enrichment resulted in higher recovery of Spn (+10.4 %, P<0.001), Hflu (+4.4 %, P=0.39) and Mcat (+13.5 %, <0.001). Broth-enrichment significantly reduces the accurate detection of bacterial respiratory pathogens and increases identification of SA and AHS in NW. Broth-enrichment improves detection of bacterial respiratory pathogens in MEF samples.

Analysis of the bacterial strains using Biolog plates in the contaminated soil from Riyadh community.

PubMed

Al-Dhabaan, Fahad Abdullah M; Bakhali, Ali Hassan

2017-05-01

Routine manufacture, detonation and disposal of explosives in land and groundwater have resulted in complete pollution. Explosives are xenobiotic compounds, being toxic to biological systems, and their recalcitrance leads to persistence in the environment. The methods currently used for the remediation of explosive contaminated sites are expensive and can result in the formation of toxic products. The present study aimed to investigate the bacterial strains using the Biolog plates in the soil from the Riyadh community. The microbial strains were isolated using the spread plate technique and were identified using the Biolog method. In this study we have analyzed from bacterial families of soil samples, obtained from the different sites in 5 regions at Explosive Institute. Our results conclude that Biolog MicroPlates were developed for the rapid identification of bacterial isolates by sole-carbon source utilization and can be used for the identification of bacteria. Out of five communities, only four families of bacteria indicate that the microbial community lacks significant diversity in region one from the Riyadh community in Saudi Arabia. More studies are needed to be carried out in different regions to validate our results.

Experimental Study for Automatic Colony Counting System Based Onimage Processing

NASA Astrophysics Data System (ADS)

Fang, Junlong; Li, Wenzhe; Wang, Guoxin

Colony counting in many colony experiments is detected by manual method at present, therefore it is difficult for man to execute the method quickly and accurately .A new automatic colony counting system was developed. Making use of image-processing technology, a study was made on the feasibility of distinguishing objectively white bacterial colonies from clear plates according to the RGB color theory. An optimal chromatic value was obtained based upon a lot of experiments on the distribution of the chromatic value. It has been proved that the method greatly improves the accuracy and efficiency of the colony counting and the counting result is not affected by using inoculation, shape or size of the colony. It is revealed that automatic detection of colony quantity using image-processing technology could be an effective way.

Impact of different-sized laminar air flow versus no laminar air flow on bacterial counts in the operating room during orthopedic surgery.

PubMed

Diab-Elschahawi, Magda; Berger, Jutta; Blacky, Alexander; Kimberger, Oliver; Oguz, Ruken; Kuelpmann, Ruediger; Kramer, Axel; Assadian, Ojan

2011-09-01

This study investigated the influence of the size of unidirectional ceiling distribution systems on counts of viable microorganisms recovered at defined sites in operating room (ORs) and on instrument tables during orthopedic surgery. We compared bacterial sedimentation during 80 orthopedic surgeries. A total of 19 surgeries were performed in ORs with a large (518 cm × 380 cm) unidirectional ceiling distribution (colloquially known as laminar air flow [LAF]) ventilation system, 21 procedures in ORs with a small (380 cm × 120 cm) LAF system, and 40 procedures in ORs with no LAF system. Bacterial sedimentation was evaluated using both settle plates and nitrocellulose membranes. Multivariate linear regression analysis revealed that the colony-forming unit count on nitrocellulose membranes positioned on the instrument table was significantly associated only with the size of the unidirectional LAF distribution system (P < .001), not with the duration of the surgical intervention (P = .753) or with the number of persons present during the surgical intervention (P = .291). Our findings indicate that simply having an LAF ventilation system in place will not provide bacteria-free conditions at the surgical site and on the instrument table. In view of the limited number of procedures studied, our findings require confirmation and further investigations on the ideal, but affordable, size of LAF ventilation systems. Copyright © 2011 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

A rapid and universal bacteria-counting approach using CdSe/ZnS/SiO2 composite nanoparticles as fluorescence probe.

PubMed

Fu, Xin; Huang, Kelong; Liu, Suqin

2010-02-01

In this paper, a rapid, simple, and sensitive method was described for detection of the total bacterial count using SiO(2)-coated CdSe/ZnS quantum dots (QDs) as a fluorescence marker that covalently coupled with bacteria using glutaraldehyde as the crosslinker. Highly luminescent CdSe/ZnS were prepared by applying cadmium oxide and zinc stearate as precursors instead of pyrophoric organometallic precursors. A reverse-microemulsion technique was used to synthesize CdSe/ZnS/SiO(2) composite nanoparticles with a SiO(2) surface coating. Our results showed that CdSe/ZnS/SiO(2) composite nanoparticles prepared with this method possessed highly luminescent, biologically functional, and monodispersive characteristics, and could successfully be covalently conjugated with the bacteria. As a demonstration, it was found that the method had higher sensitivity and could count bacteria in 3 x 10(2) CFU/mL, lower than the conventional plate counting and organic dye-based method. A linear relationship of the fluorescence peak intensity (Y) and the total bacterial count (X) was established in the range of 3 x 10(2)-10(7) CFU/mL using the equation Y = 374.82X-938.27 (R = 0.99574). The results of the determination for the total count of bacteria in seven real samples were identical with the conventional plate count method, and the standard deviation was satisfactory.

In vitro effect of Reiki treatment on bacterial cultures: Role of experimental context and practitioner well-being.

PubMed

Rubik, Beverly; Brooks, Audrey J; Schwartz, Gary E

2006-01-01

To measure effects of Reiki treatments on growth of heat-shocked bacteria, and to determine the influence of healing context and practitioner well-being. Overnight cultures of Escherichia coli K12 in fresh medium were used. Culture samples were paired with controls to minimize any ordering effects. Samples were heat-shocked prior to Reiki treatment, which was performed by Reiki practitioners for up to 15 minutes, with untreated controls. Plate-count assay using an automated colony counter determined the number of viable bacteria. Fourteen Reiki practitioners each completed 3 runs (n = 42 runs) without healing context, and another 2 runs (n = 28 runs) in which they first treated a pain patient for 30 minutes (healing context). Well-being questionnaires were administered to practitioners pre-post all sessions. No overall difference was found between the Reiki and control plates in the nonhealing context. In the healing context, the Reiki treated cultures overall exhibited significantly more bacteria than controls (p < 0.05). Practitioner social (p < 0.013) and emotional well-being (p < 0.021) correlated with Reiki treatment outcome on bacterial cultures in the nonhealing context. Practitioner social (p < 0.031), physical (p < 0.030), and emotional (p < 0.026) well-being correlated with Reiki treatment outcome on the bacterial cultures in the healing context. For practitioners starting with diminished well-being, control counts were likely to be higher than Reiki-treated bacterial counts. For practitioners starting with a higher level of well-being, Reiki counts were likely to be higher than control counts. Reiki improved growth of heat-shocked bacterial cultures in a healing context. The initial level of well-being of the Reiki practitioners correlates with the outcome of Reiki on bacterial culture growth and is key to the results obtained.

Effects of bacterial pollution caused by a strong typhoon event and the restoration of a recreational beach: Transitions of fecal bacterial counts and bacterial flora in beach sand.

PubMed

Suzuki, Yoshihiro; Teranishi, Kotaro; Matsuwaki, Tomonori; Nukazawa, Kei; Ogura, Yoshitoshi

2018-05-28

To determine the effects of bacteria pollution associated with a strong typhoon event and to assess the restoration of the normal bacterial flora, we used conventional filtration methods and nextgeneration sequencing of 16S rRNA genes to analyze the transition of fecal and total bacterial counts in water and core sand samples collected from a recreational beach. Immediately after the typhoon event, Escherichia coli counts increased to 82 CFU/100 g in the surface beach sand. E. coli was detected through the surface to sand 85-cm deep at the land side point (10-m land side from the high-water line). However, E. coli disappeared within a month from the land side point. The composition of the bacterial flora in the beach sand at the land point was directly influenced by the typhoon event. Pseudomonas was the most prevalent genus throughout the sand layers (0-102-cm deep) during the typhoon event. After 3 months, the population of Pseudomonas significantly decreased, and the predominant genus in the surface layer was Kaistobacter, although Pseudomonas was the major genus in the 17- to 85-cm layer. When the beach conditions stabilized, the number of pollutant Pseudomonas among the 10 most abundant genera decreased to lower than the limit of detection. The bacterial population of the sand was subsequently restored to the most populous pre-event orders at the land point. A land-side beach, where users directly contact the sand, was significantly affected by bacterial pollution caused by a strong typhoon event. We show here that the normal bacterial flora of the surface sand was restored within 1 month. Copyright © 2018 Elsevier B.V. All rights reserved.

Two-dimensional photon-counting detector arrays based on microchannel array plates

NASA Technical Reports Server (NTRS)

Timothy, J. G.; Bybee, R. L.

1975-01-01

The production of simple and rugged photon-counting detector arrays has been made possible by recent improvements in the performance of the microchannel array plate (MCP) and by the parallel development of compatible electronic readout systems. The construction of proximity-focused MCP arrays of novel design in which photometric information from (n x m) picture elements is read out with a total of (n + m) amplifier and discriminator circuits is described. Results obtained with a breadboard (32 x 32)-element array employing 64 charge-sensitive amplifiers are presented, and the application of systems of this type in spectrometers and cameras for use with ground-based telescopes and on orbiting spacecraft discussed.

Use of simulation tools to illustrate the effect of data management practices for low and negative plate counts on the estimated parameters of microbial reduction models.

PubMed

Garcés-Vega, Francisco; Marks, Bradley P

2014-08-01

In the last 20 years, the use of microbial reduction models has expanded significantly, including inactivation (linear and nonlinear), survival, and transfer models. However, a major constraint for model development is the impossibility to directly quantify the number of viable microorganisms below the limit of detection (LOD) for a given study. Different approaches have been used to manage this challenge, including ignoring negative plate counts, using statistical estimations, or applying data transformations. Our objective was to illustrate and quantify the effect of negative plate count data management approaches on parameter estimation for microbial reduction models. Because it is impossible to obtain accurate plate counts below the LOD, we performed simulated experiments to generate synthetic data for both log-linear and Weibull-type microbial reductions. We then applied five different, previously reported data management practices and fit log-linear and Weibull models to the resulting data. The results indicated a significant effect (α = 0.05) of the data management practices on the estimated model parameters and performance indicators. For example, when the negative plate counts were replaced by the LOD for log-linear data sets, the slope of the subsequent log-linear model was, on average, 22% smaller than for the original data, the resulting model underpredicted lethality by up to 2.0 log, and the Weibull model was erroneously selected as the most likely correct model for those data. The results demonstrate that it is important to explicitly report LODs and related data management protocols, which can significantly affect model results, interpretation, and utility. Ultimately, we recommend using only the positive plate counts to estimate model parameters for microbial reduction curves and avoiding any data value substitutions or transformations when managing negative plate counts to yield the most accurate model parameters.

Automated agar plate streaker: a linear plater on Society for Biomolecular Sciences standard plates.

PubMed

King, Gregory W; Kath, Gary S; Siciliano, Sal; Simpson, Neal; Masurekar, Prakash; Sigmund, Jan; Polishook, Jon; Skwish, Stephen; Bills, Gerald; Genilloud, Olga; Peláez, Fernando; Martín, Jesus; Dufresne, Claude

2006-09-01

Several protocols for bacterial isolation and techniques for aerobic plate counting rely on the use of a spiral plater to deposit concentration gradients of microbial suspensions onto a circular agar plate to isolate colony growth. The advantage of applying a gradient of concentrations across the agar surface is that the original microbiological sample can be applied at a single concentration rather than as multiple serial dilutions. The spiral plater gradually dilutes the sample across a compact area and therefore saves time preparing dilutions and multiple agar plates. Commercial spiral platers are not automated and require manual sample loading. Dispensing of the sample volume and rate of gradients are often very limited in range. Furthermore, the spiral sample application cannot be used with rectangular microplates. Another limitation of commercial spiral platers is that they are useful only for dilute, filtered suspensions and cannot plate suspensions of coarse organic particles therefore precluding the use of many kinds of microorganism-containing substrata. An automated agar plate spreader capable of processing 99 rectangular microplates in unattended mode is described. This novel instrument is capable of dispensing discrete volumes of sample in a linear pattern. It can be programmed to dispense a sample suspense at a uniform application rate or across a decreasing concentration gradient.

Anatomical characteristics of teats and premilking bacterial counts of teat skin swabs of primiparous cows exposed to different types of bedding.

PubMed

Guarín, J F; Baumberger, C; Ruegg, P L

2017-02-01

Bacterial populations of teat skin are associated with risk of intramammary infection and may be influenced by anatomical characteristics of teats. The objective of this study was to evaluate associations of selected anatomical characteristics of teats with bacterial counts of teat skin of cows exposed to different types of bedding. Primarily primiparous Holstein cows (n = 128) were randomly allocated to 4 pens within a single barn. Each pen contained 1 type of bedding [new sand (NES), recycled sand (RS), deep-bedded manure solids (DBMS), and shallow-bedded manure solids over foam core mattresses (SBMS)]. During a single farm visit udders (n = 112) were scored for hygiene and 1 front (n = 112) and 1 rear teat (n = 111) of each enrolled cow were scored for hyperkeratosis (HK). Teat length, teat barrel diameter, and teat apex diameter were measured and teat skin swabs were systematically collected for microbiological analysis. Linear type evaluation data for udders of each cow were retrieved for each cow. Teat position (front or rear) was associated with occurrence of clinical mastitis during the 12 mo before the farm visit and more cases occurred in front quarters. The proportion of udders that were classified as clean (score 1 or 2) was 68, 82, 54, and 95% for cows housed in pens containing NES, RS, SBMS, and DBMS, respectively. No association was found between HK score and teat position and no association was found between HK score and teat skin bacterial count. Bacterial counts of teat skin swabs from front teats of cows in pens containing RS and SBMS were significantly less than those of rear teats of cows in pens containing DBMS or NES. Teat skin bacterial counts were significantly greater for swabs obtained from teats of cows with udder hygiene scores of 3 and 4 as compared with swabs obtained from cows with cleaner udders. Of all udder conformation traits evaluated, only narrower rear teat placement was positively associated with bacterial counts on teat skin

Comparison of Primary Models to Predict Microbial Growth by the Plate Count and Absorbance Methods.

PubMed

Pla, María-Leonor; Oltra, Sandra; Esteban, María-Dolores; Andreu, Santiago; Palop, Alfredo

2015-01-01

The selection of a primary model to describe microbial growth in predictive food microbiology often appears to be subjective. The objective of this research was to check the performance of different mathematical models in predicting growth parameters, both by absorbance and plate count methods. For this purpose, growth curves of three different microorganisms (Bacillus cereus, Listeria monocytogenes, and Escherichia coli) grown under the same conditions, but with different initial concentrations each, were analysed. When measuring the microbial growth of each microorganism by optical density, almost all models provided quite high goodness of fit (r(2) > 0.93) for all growth curves. The growth rate remained approximately constant for all growth curves of each microorganism, when considering one growth model, but differences were found among models. Three-phase linear model provided the lowest variation for growth rate values for all three microorganisms. Baranyi model gave a variation marginally higher, despite a much better overall fitting. When measuring the microbial growth by plate count, similar results were obtained. These results provide insight into predictive microbiology and will help food microbiologists and researchers to choose the proper primary growth predictive model.

Small-Scale Vertical Distribution of Bacterial Biomass and Diversity in Biological Soil Crusts from Arid Lands in the Colorado Plateau

USGS Publications Warehouse

Garcia-Pichel, F.; Johnson, S.L.; Youngkin, D.; Belnap, J.

2003-01-01

We characterized, at millimeter resolution, bacterial biomass, diversity, and vertical stratification of biological soil crusts in arid lands from the Colorado Plateau. Microscopic counts, extractable DNA, and plate counts of viable aerobic copiotrophs (VAC) revealed that the top centimeter of crusted soils contained atypically large bacterial populations, tenfold larger than those in uncrusted, deeper soils. The plate counts were not always consistent with more direct estimates of microbial biomass. Bacterial populations peaked at the immediate subsurface (1-2 mm) in light-appearing, young crusts, and at the surface (0-1 mm) in well-developed, dark crusts, which corresponds to the location of cyanobacterial populations. Bacterial abundance decreased with depth below these horizons. Spatially resolved DGGE fingerprints of Bacterial 16S rRNA genes demonstrated the presence of highly diverse natural communities, but we could detect neither trends with depth in bacterial richness or diversity, nor a difference in diversity indices between crust types. Fingerprints, however, revealed the presence of marked stratification in the structure of the microbial communities, probably a result of vertical gradients in physicochemical parameters. Sequencing and phylogenetic analyses indicated that most of the naturally occurring bacteria are novel types, with low sequence similarity (83-93%) to those available in public databases. DGGE analyses of the VAC populations indicated communities of lower diversity, with most types having sequences more than 94% similar to those in public databases. Our study indicates that soil crusts represent small-scale mantles of fertility in arid ecosystems, harboring vertically structured, little-known bacterial populations that are not well represented by standard cultivation methods.

Correlation between mastitis occurrence and the count of microorganisms in bulk raw milk of bovine dairy herds in four selective culture media.

PubMed

Souto, Luís I M; Minagawa, Clarice Y; Telles, Evelise O; Garbuglio, Márcio A; Amaku, Marcos; Melville, Priscilla A; Dias, Ricardo A; Sakata, Sonia T; Benites, Nilson R

2010-02-01

Milk is the normal secretion of the mammary gland, practically free of colostrum and obtained by the complete milking of one or more healthy animals. Mastitis is an inflammatory process of the mammary gland and it may cause alterations in the milk. The present work aimed to verify whether it is possible, by means of the counts of microorganism in the bulk raw milk in four selective culture media, to establish a correlation with the occurrence of mastitis and therefore, to monitor this disease in bovine dairy herds. The following selective culture media were used: KF Streptococcus Agar, Edwards Agar, Baird-Parker Agar, Blood Agar plus potassium tellurite. Spearman's correlation coefficient was calculated in order to compare the occurrence of mastitis (percentage) in each herd with respective selective culture media counts of microorganisms in bulk raw milk. Thirty-six possibilities were analysed (Tamis and CMT-positive rates were compared with the log-transformed count in four selective culture media) and there was a negative correlation between Tamis 3 and the Baird-Parker Agar plate count. The total results of microbiological tests showed that there were three correlations of the counts in selective culture media. Fifty-two possibilities were analysed and there was a negative correlation between no-bacterial-growth mastitis rates and log10 of KF Streptoccocus Agar plate count and there were two positive correlations between coagulase-positive staphylococci and log10 of Baird-Parker Agar plate count and Blood Agar plus potassium tellurite plate count.

Hydrodebridement of wounds: effectiveness in reducing wound bacterial contamination and potential for air bacterial contamination.

PubMed

Bowling, Frank L; Stickings, Daryl S; Edwards-Jones, Valerie; Armstrong, David G; Boulton, Andrew Jm

2009-05-08

The purpose of this study was to assess the level of air contamination with bacteria after surgical hydrodebridement and to determine the effectiveness of hydro surgery on bacterial reduction of a simulated infected wound. Four porcine samples were scored then infected with a broth culture containing a variety of organisms and incubated at 37 degrees C for 24 hours. The infected samples were then debrided with the hydro surgery tool (Versajet, Smith and Nephew, Largo, Florida, USA). Samples were taken for microbiology, histology and scanning electron microscopy pre-infection, post infection and post debridement. Air bacterial contamination was evaluated before, during and after debridement by using active and passive methods; for active sampling the SAS-Super 90 air sampler was used, for passive sampling settle plates were located at set distances around the clinic room. There was no statistically significant reduction in bacterial contamination of the porcine samples post hydrodebridement. Analysis of the passive sampling showed a significant (p < 0.001) increase in microbial counts post hydrodebridement. Levels ranging from 950 colony forming units per meter cubed (CFUs/m3) to 16780 CFUs/m3 were observed with active sampling of the air whilst using hydro surgery equipment compared with a basal count of 582 CFUs/m3. During removal of the wound dressing, a significant increase was observed relative to basal counts (p < 0.05). Microbial load of the air samples was still significantly raised 1 hour post-therapy. The results suggest a significant increase in bacterial air contamination both by active sampling and passive sampling. We believe that action might be taken to mitigate fallout in the settings in which this technique is used.

Optimization of single plate-serial dilution spotting (SP-SDS) with sample anchoring as an assured method for bacterial and yeast cfu enumeration and single colony isolation from diverse samples.

PubMed

Thomas, Pious; Sekhar, Aparna C; Upreti, Reshmi; Mujawar, Mohammad M; Pasha, Sadiq S

2015-12-01

We propose a simple technique for bacterial and yeast cfu estimations from diverse samples with no prior idea of viable counts, designated as single plate-serial dilution spotting (SP-SDS) with the prime recommendation of sample anchoring (10 0 stocks). For pure cultures, serial dilutions were prepared from 0.1 OD (10 0 ) stock and 20 μl aliquots of six dilutions (10 1 -10 6 ) were applied as 10-15 micro-drops in six sectors over agar-gelled medium in 9-cm plates. For liquid samples 10 0 -10 5 dilutions, and for colloidal suspensions and solid samples (10% w/v), 10 1 -10 6 dilutions were used. Following incubation, at least one dilution level yielded 6-60 cfu per sector comparable to the standard method involving 100 μl samples. Tested on diverse bacteria, composite samples and Saccharomyces cerevisiae , SP-SDS offered wider applicability over alternative methods like drop-plating and track-dilution for cfu estimation, single colony isolation and culture purity testing, particularly suiting low resource settings.

Heterotrophic plate count and consumer's health under special consideration of water softeners.

PubMed

Hambsch, Beate; Sacré, Clara; Wagner, Ivo

2004-05-01

The phenomenon of bacterial growth in water softeners is well known since years. To upgrade the hygienic safety of water softeners, the German DIN Standard 19636 was developed, to assure that the distribution system could not be contaminated by these devices and that the drinking water to be used in the household still meets the microbiological standards according to the German drinking water guidelines, i.e. among others heterotrophic plate count (HPC) below 100 CFU/ml. Moreover, the standard for the water softeners includes a test for contamination with Pseudomonas aeruginosa which has to be disinfected during the regeneration phase. This is possible by sanitizing the resin bed during regeneration by producing chlorine. The results of the last 10 years of tests of water softeners according to DIN 19636 showed that it is possible to produce water softeners that comply with that standard. Approximately 60% of the tested models were accepted. P. aeruginosa is used as an indicator for potentially pathogenic bacteria being able to grow also in low nutrient conditions which normally prevail in drinking water. Like other heterotrophs, the numbers of P. aeruginosa increase rapidly as stagnation occurs. Normally P. aeruginosa is not present in the distributed drinking water. However, under certain conditions, P. aeruginosa can be introduced into the drinking water distribution system, for instance, during construction work. The occurrence of P. aeruginosa is shown in different cases in treatment plants, public drinking water systems and in-house installations. The compliance with DIN 19636 provides assurance that a water softener will not be a constant source of contamination, even if it is once inoculated with a potentially pathogenic bacterium like P. aeruginosa. Copyright 2003 Elsevier B.V.

Steam versus hot-water scalding in reducing bacterial loads on the skin of commercially processed poultry.

PubMed

Patrick, T E; Goodwin, T L; Collins, J A; Wyche, R C; Love, B E

1972-04-01

A comparison of two types of scalders was conducted to determine their effectiveness in reducing bacterial contamination of poultry carcasses. A conventional hot-water scalder and a prototype model of a steam scalder were tested under commercial conditions. Total plate counts from steam-scalded birds were significantly lower than the counts of water-scalded birds immediately after scalding and again after picking. No differences in the two methods could be found after chilling. Coliform counts from steam-scalded birds were significantly lower than the counts from water-scalded birds immediately after scalding. No significant differences in coliform counts were detected when the two scald methods were compared after defeathering and chilling.

Responses of Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus to Simulated Food Processing Treatments, Determined Using Fluorescence-Activated Cell Sorting and Plate Counting▿

PubMed Central

Kennedy, Deirdre; Cronin, Ultan P.; Wilkinson, Martin G.

2011-01-01

Three common food pathogenic microorganisms were exposed to treatments simulating those used in food processing. Treated cell suspensions were then analyzed for reduction in growth by plate counting. Flow cytometry (FCM) and fluorescence-activated cell sorting (FACS) were carried out on treated cells stained for membrane integrity (Syto 9/propidium iodide) or the presence of membrane potential [DiOC2(3)]. For each microbial species, representative cells from various subpopulations detected by FCM were sorted onto selective and nonselective agar and evaluated for growth and recovery rates. In general, treatments giving rise to the highest reductions in counts also had the greatest effects on cell membrane integrity and membrane potential. Overall, treatments that impacted cell membrane permeability did not necessarily have a comparable effect on membrane potential. In addition, some bacterial species with extensively damaged membranes, as detected by FCM, appeared to be able to replicate and grow after sorting. Growth of sorted cells from various subpopulations was not always reflected in plate counts, and in some cases the staining protocol may have rendered cells unculturable. Optimized FCM protocols generated a greater insight into the extent of the heterogeneous bacterial population responses to food control measures than did plate counts. This study underlined the requirement to use FACS to relate various cytometric profiles generated by various staining protocols with the ability of cells to grow on microbial agar plates. Such information is a prerequisite for more-widespread adoption of FCM as a routine microbiological analytical technique. PMID:21602370

Determination of viability of Aeromonas hydrophila in increasing concentrations of sodium chloride at different temperatures by flow cytometry and plate count technique.

PubMed

Pianetti, Anna; Manti, Anita; Boi, Paola; Citterio, Barbara; Sabatini, Luigia; Papa, Stefano; Rocchi, Marco Bruno Luigi; Bruscolini, Francesca

2008-10-31

Aeromonads in waters and foods can represent a risk to human health. Factors such as sodium chloride concentration and temperature can affect growth and viability of several food and water-borne pathogens. The behaviour of an Aeromonas hydrophila strain in the presence of 1.7%, 3.4% and 6% NaCl concentrations at 24 degrees C and 4 degrees C was studied over a 188 day period. Viability and membrane potential were assessed by flow cytometry; growth was evaluated by plate count technique. Flow cytometry evidenced that A. hydrophila retained viability over the period although varying according to temperature and salt concentrations. Colony Forming Units were generally lower in number than viable cells especially in the presence of 6% NaCl, indicating the occurrence of stressed cells which maintain metabolic activity yet are not able to grow on agar plates. In conclusion, A. hydrophila showed a long-term halotolerance even at elevated (6%) NaCl concentrations and a lesser sensitivity to salt at low temperature; therefore, low temperature and salt, which are two important factors limiting bacterial growth, do not assure safety in the case of high initial contamination. Finally, cytometry appears a valid tool for the rapid detection of the viability of pathogenic bacteria in food and environmental matrices to control and prevent health risks.

Selective enumeration of propionibacteria in Emmental-type cheese using Petrifilm™ aerobic count plates added to lithium glycerol broth.

PubMed

de Freitas, Rosângela; Luiz, Lívia M Pinheiro; Alves, Maura Pinheiro; Valence-Bertel, Florence; Nero, Luís Augusto; de Carvalho, Antônio Fernandes

2013-08-01

Propionibacteria derived from dairy products are relevant starter cultures for the production of Swiss and Emmental-type cheeses, and the monitoring of which is mandatory for proper quality control. This study aimed to evaluate an alternative procedure to enumerate propionibacteria, in order to develop a reliable and practical methodology to be employed by dairy industries. 2,3,5-triphenyltetrazolium chloride (TTC) inhibitory activity was tested against five reference strains (CIRM 09, 38, 39, 40 and 116); TTC at 0·0025% (w/v) was not inhibitory, with the exception of one strain (CIRM 116). Subsequently, the four TTC-resistant strains, three commercial starter cultures (PS-1, PB-I, and CHOO) and twelve Emmental-type cheese samples were subjected to propionibacteria enumeration using Lithium Glycerol (LG) agar, and Petrifilm™ Aerobic Count (AC) plates added to LG broth (anaerobic incubation at 30 °C for 7 d). Petrifilm™ AC added to LG broth presented high counts than LG agar (P<0·05) for only two reference strains (CIRM 39, and 40) and for all commercial starter cultures. Cheese sample counts obtained by both procedures did not show significant differences (P<0·05). Significant correlation indexes were observed between the counts recorded by both methods (P<0·05). These results demonstrate the reliability of Petrifilm™ AC plates added to LG broth in enumerating select Propionibacterium spp., despite some limitations observed for specific commercial starter cultures.

Studies on stannous fluoride toothpaste and gel (2). Effects on salivary bacterial counts and plaque regrowth in vivo.

PubMed

Addy, M; Greenman, J; Renton-Harper, P; Newcombe, R; Doherty, F

1997-02-01

There has been a resurgence of interest in stannous fluoride (SF) products in particular to provide oral hygiene and gingival health benefits. The aim of this study was to assess the persistence of antimicrobial action of a number of SF formulations in the mouth and relate these to plaque inhibitory activity. The formulations were 2 SF toothpastes (SF1, SF2), 2 SF plus stannous pyrophosphate toothpastes (SFSP1, SFSP2), a SF gel (G), a NaF toothpaste (C) and saline (S) as control. Both studies involve 2 different groups of 21 healthy dentate volunteers. The studies were single, blind, randomised, crossover designs balanced for residual effects, with a minimum 2 1/2 day washout period. Salivary bacterial counts were determined before and to 7 h after a single rinse with the formulations. Plaque regrowth from a zero baseline (day 1) was measured by index and area on day 5, after 2x daily rinsing with slurries of the formulations or saline. For bacterial counts, highly significant treatment differences were found. Bacterial counts were variably reduced by all treatments to 30 min then showed a variable rate of return towards baseline. All test agents were significantly better than S at some timepoints. The order for greatest persistence of action downwards was; (1) SFSP2; (2) SFSP1, G, and SF1; (3) SF2; (4) C; (5) S. Highly significant differences in plaque regrowth between treatments were found with similar mean ordering of efficacy as for salivary bacterial counts from most effective downwards namely; (1) SFSP1 and SFSP2; (2) SF1; (3) SF2; G and C; (4) S. The results were consistent with a parallel study measuring tea staining in vitro, whereby formulations causing the most staining produced the greatest persistence of action and plaque inhibitory activity. This suggests the availability of stannous ions was important for the clinical effects. It is concluded that stannous ions can enhance the plaque inhibitory action of toothpaste via a persistent antimicrobial

Method for Performing Aerobic Plate Counts of Anhydrous Cosmetics Utilizing Tween 60 and Arlacel 80 as Dispersing Agents

PubMed Central

McConville, John F.; Anger, Claude B.; Anderson, David W.

1974-01-01

An aqueous diluent containing Tween 60 and Arlacel 80 gave greater recovery of microorganisms when compared with two common diluents as determined by aerobic plate count of inoculated anhydrous cosmetics. The greater recovery was caused by better dispersion of the anhydrous cosmetics in the diluents. Images PMID:4203790

Microcoupon Assay Of Adhesion And Growth Of Bacterial Films

NASA Technical Reports Server (NTRS)

Pierson, Duane L.; Koenig, David W.

1994-01-01

Microbiological assay technique facilitates determination of some characteristics of sessile bacteria like those that attach to and coat interior walls of water-purification systems. Biofilms cause sickness and interfere with purification process. Technique enables direct measurement of rate of attachment of bacterial cells, their metabolism, and effects of chemicals on them. Used to quantify effects of both bactericides and growth-stimulating agents and in place of older standard plate-count and tube-dilution techniques.

Bare below elbows: does this policy affect handwashing efficacy and reduce bacterial colonisation?

PubMed Central

Burger, A; Wijewardena, C; Clayson, S; Greatorex, RA

2010-01-01

INTRODUCTION UK Department of Health guidelines recommend that clinical staff are ‘bare below the elbows’. There is a paucity of evidence to support this policy. One may hypothesise that absence of clothing around wrists facilitates more effective handwashing: this study aims to establish whether dress code affects bacterial colonisation before and after handwashing. SUBJECTS AND METHODS Sixty-six clinical staff volunteered to take part in the study, noting whether they were bare below the elbows (BBE) or not bare (NB). Using a standardised technique, imprints of left and right fingers, palms, wrists and forearms were taken onto mini agar plates. Imprints were repeated after handwashing. After incubation, colonies per plate were counted, and subcultures taken. RESULTS Thirty-eight staff were BBE and 28 were not. A total of 1112 plates were cultured. Before handwashing there was no significant difference in number of colonies between BBE and NB groups (Mann–Whitney, P < 0.05). Handwashing reduced the colony count, with greatest effect on fingers, palms and dominant wrists (t-test, P < 0.05). Comparing the two groups again after handwashing revealed no significant difference (Mann–Whitney, P < 0.05). Subcultures revealed predominantly skin flora. CONCLUSIONS There was a large variation in number of colonies cultured. Handwashing resulted in a statistically significant reduction in colony count on fingers, palms and dominant wrist regardless of clothing. We conclude that handwashing produces a significant reduction in number of bacterial colonies on staff hands, and that clothing that is not BBE does not impede this reduction. PMID:20727253

Precise, High-throughput Analysis of Bacterial Growth.

PubMed

Kurokawa, Masaomi; Ying, Bei-Wen

2017-09-19

Bacterial growth is a central concept in the development of modern microbial physiology, as well as in the investigation of cellular dynamics at the systems level. Recent studies have reported correlations between bacterial growth and genome-wide events, such as genome reduction and transcriptome reorganization. Correctly analyzing bacterial growth is crucial for understanding the growth-dependent coordination of gene functions and cellular components. Accordingly, the precise quantitative evaluation of bacterial growth in a high-throughput manner is required. Emerging technological developments offer new experimental tools that allow updates of the methods used for studying bacterial growth. The protocol introduced here employs a microplate reader with a highly optimized experimental procedure for the reproducible and precise evaluation of bacterial growth. This protocol was used to evaluate the growth of several previously described Escherichia coli strains. The main steps of the protocol are as follows: the preparation of a large number of cell stocks in small vials for repeated tests with reproducible results, the use of 96-well plates for high-throughput growth evaluation, and the manual calculation of two major parameters (i.e., maximal growth rate and population density) representing the growth dynamics. In comparison to the traditional colony-forming unit (CFU) assay, which counts the cells that are cultured in glass tubes over time on agar plates, the present method is more efficient and provides more detailed temporal records of growth changes, but has a stricter detection limit at low population densities. In summary, the described method is advantageous for the precise and reproducible high-throughput analysis of bacterial growth, which can be used to draw conceptual conclusions or to make theoretical observations.

Aseptic laboratory techniques: plating methods.

PubMed

Sanders, Erin R

2012-05-11

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: Perform plating procedures without contaminating media. Isolate single bacterial colonies by the streak-plating

Aseptic Laboratory Techniques: Plating Methods

PubMed Central

Sanders, Erin R.

2012-01-01

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: ● Perform plating procedures without contaminating media. ● Isolate single bacterial colonies by the

Steam Versus Hot-Water Scalding in Reducing Bacterial Loads on the Skin of Commercially Processed Poultry

PubMed Central

Patrick, Thomas E.; Goodwin, T. L.; Collins, J. A.; Wyche, R. C.; Love, B. E.

1972-01-01

A comparison of two types of scalders was conducted to determine their effectiveness in reducing bacterial contamination of poultry carcasses. A conventional hot-water scalder and a prototype model of a steam scalder were tested under commercial conditions. Total plate counts from steam-scalded birds were significantly lower than the counts of water-scalded birds immediately after scalding and again after picking. No differences in the two methods could be found after chilling. Coliform counts from steam-scalded birds were significantly lower than the counts from water-scalded birds immediately after scalding. No significant differences in coliform counts were detected when the two scald methods were compared after defeathering and chilling. PMID:4553146

Chromophore-enhanced bacterial photothermolysis

NASA Astrophysics Data System (ADS)

Huckleby, Jana K.; Morton, Rebecca J.; Bartels, Kenneth E.

1999-06-01

The use of chromophore dyes to enhance the bactericidal effect of laser energy was studied as a means to optimize laser treatment for the decontamination of wound. Using an in vitro study, various concentrations of indocyanine green (ICG), carbon black, and fluorescein were mixed with a suspension of bacteria and plated on tryptic soy agar. Plates were exposed to a laser beam of 10-15 watts for times ranging from 0 to 180 seconds, incubated overnight, and colony counts were performed. Bacteria not mixed with chromophore were used as controls. Six bacterial strains encompassing a range of bacterial types were used: Staphylococcus aureau, Streptococcus pyogenes, Escherichia coli, Pseudomonas aeruginosa, Bacillus cereus spore suspensions, and Clostridium perfringens. Laser treatment alone had no effect on any of the bacteria. Significant killing of gram-positive bacteria, including spores of Bacillus cereus, was observed only with the use of ICG and diode laser energy. No effect was observed using any of the chromophores on the gram-negative bacteria. The results of this study indicate that successful killing of gram-positive bacteria can be achieved using ICG combined with appropriate laser energy and wavelength. Efforts to enhance the susceptibility of gram-negative bacteria to photothermolysis by laser energy were unsuccessful.

Correlation between standard plate count and somatic cell count milk quality results for Wisconsin dairy producers.

PubMed

Borneman, Darand L; Ingham, Steve

2014-05-01

The objective of this study was to determine if a correlation exists between standard plate count (SPC) and somatic cell count (SCC) monthly reported results for Wisconsin dairy producers. Such a correlation may indicate that Wisconsin producers effectively controlling sanitation and milk temperature (reflected in low SPC) also have implemented good herd health management practices (reflected in low SCC). The SPC and SCC results for all grade A and B dairy producers who submitted results to the Wisconsin Department of Agriculture, Trade, and Consumer Protection, in each month of 2012 were analyzed. Grade A producer SPC results were less dispersed than grade B producer SPC results. Regression analysis showed a highly significant correlation between SPC and SCC, but the R(2) value was very small (0.02-0.03), suggesting that many other factors, besides SCC, influence SPC. Average SCC (across 12 mo) for grade A and B producers decreased with an increase in the number of monthly SPC results (out of 12) that were ≤ 25,000 cfu/mL. A chi-squared test of independence showed that the proportion of monthly SCC results >250,000 cells/mL varied significantly depending on whether the corresponding SPC result was ≤ 25,000 or >25,000 cfu/mL. This significant difference occurred in all months of 2012 for grade A and B producers. The results suggest that a generally consistent level of skill exists across dairy production practices affecting SPC and SCC. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Development and test of photon-counting microchannel plate detector arrays for use on space telescopes

NASA Technical Reports Server (NTRS)

Timothy, J. G.

1976-01-01

The full sensitivity, dynamic range, and photometric stability of microchannel array plates(MCP) are incorporated into a photon-counting detection system for space operations. Components of the system include feedback-free MCP's for high gain and saturated output pulse-height distribution with a stable response; multi-anode readout arrays mounted in proximity focus with the output face of the MCP; and multi-layer ceramic headers to provide electrical interface between the anode array in a sealed detector tube and the associated electronics.

A miniaturized counting technique for anaerobic bacteria.

PubMed

Sharpe, A N; Pettipher, G L; Lloyd, G R

1976-12-01

A miniaturized counting technique gave results as good as the pour-plate and Most Probable Number (MPN) techniques for enumeration of clostridia spp. and anaerobic isolates from the gut. Highest counts were obtained when ascorbic acid (1%) and dithiothreitol (0.015%) were added to the reinforced clostridial medium used for counting. This minimized the effect of exposure to air before incubation. The miniature technique allowed up to 40 samples to be plated and incubated in one McIntosh-Filde's-type anaerobic jar, compared with 3 or 4 by the normal pour plate.

PCR-DGGE analysis of bacterial community dynamics in kava beverages during refrigeration.

PubMed

Dong, J; Kandukuru, P; Huang, A S; Li, Y

2011-07-01

Kava beverages are highly perishable even under refrigerated conditions. This study aimed to investigate the bacterial community dynamics in kava beverages during refrigeration.  Four freshly made kava beverages were obtained from kava bars and stored at 4°C. On days 0, 3 and 6, the aerobic plate count (APC), lactic acid bacteria (LAB) count and yeast and mould count (YMC) of the samples were determined. Meanwhile, bacterial DNA was extracted from each sample and subjected to the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Moreover, species-specific PCR assays were employed to identify predominant Pseudomonas spp. involved in kava spoilage. Over the storage period, the APC, LAB count and YMC of the four kava beverages all increased, whereas their pH values decreased. The DGGE profile revealed diverse bacterial populations in the samples. LAB, such as Weissella soli, Lactobacillus spp. and Lactococcus lactis, were found in the kava beverages. Species-specific PCR assays detected Pseudomonas putida and Pseudomonas fluorescens in the samples; Ps. fluorescens became dominant during refrigeration. LAB and Pseudomonas may play a significant role in the spoilage of kava beverages. This study provides important information that may be used to extend the shelf life of kava beverages. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

Comparison of bulk-tank standard plate count and somatic cell count for Wisconsin dairy farms in three size categories.

PubMed

Ingham, S C; Hu, Y; Ané, C

2011-08-01

The objective of this study was to evaluate possible claims by advocates of small-scale dairy farming that milk from smaller Wisconsin farms is of higher quality than milk from larger Wisconsin farms. Reported bulk tank standard plate count (SPC) and somatic cell count (SCC) test results for Wisconsin dairy farms were obtained for February to December, 2008. Farms were sorted into 3 size categories using available size-tracking criteria: small (≤118 cows; 12,866 farms), large (119-713 cattle; 1,565 farms), and confined animal feeding operations (≥714 cattle; 160 farms). Group means were calculated (group=farm size category) for the farms' minimum, median, mean, 90th percentile, and maximum SPC and SCC. Statistical analysis showed that group means for median, mean, 90th percentile, and maximum SPC and SCC were almost always significantly higher for the small farm category than for the large farm and confined animal feeding operations farm categories. With SPC and SCC as quality criteria and the 3 farm size categories of ≤118, 119 to 713, and ≥714 cattle, the claim of Wisconsin smaller farms producing higher quality milk than Wisconsin larger farms cannot be supported. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Potentially pathogenic features of heterotrophic plate count bacteria isolated from treated and untreated drinking water.

PubMed

Pavlov, D; de Wet, C M E; Grabow, W O K; Ehlers, M M

2004-05-01

Heterotrophic plate counts (HPCs) are commonly used to assess the general microbiological quality of drinking water. Drinking water quality specifications worldwide recommend HPC limits from 100 to 500 cfu ml(-1). A number of recent studies revealed evidence that these bacteria may not be as harmless as generally accepted. It appears that immuno-compromised individuals are particularly at risk. This would include the very young and very old patients with diseases such as AIDS and patients on therapy for purposes such as organ transplantation and cancer treatment. In this study, 339 bacterial colonies were isolated at random from selected treated and untreated drinking water in South Africa using routine heterotrophic plate count tests. In a first step to screen for potentially pathogenic properties, 188 (55.5%) of the isolates showed alpha- or beta-haemolysis on human- and horse-blood agar media. Subsequent analysis of the haemolytic isolates for enzymatic properties associated with pathogenicity revealed the presence of chondroitinase in 5.3% of the isolates, coagulase in 16.0%, DNase in 60.6%, elastase in 33.0%, fibrinolysin in 53.7%, gelatinase in 62.2%, hyaluronidase in 21.3%, lecithinase in 47.9%, lipase in 54.8% and proteinase in 64.4%. Fluorescein and pyocyanin were not produced by any of the isolates. Among the haemolytic isolates, 77.7% were resistant to oxacillin 1 microg, 59.6% to penicillin G 2 units, 47.3% to penicillin G 10 units, 54.3% to ampicillin 10 microg and 43.1% to ampicillin 25 microg. Cell culture studies revealed that 96% of haemolytic isolates were cytotoxic to HEp-2 cells, and 98.9% of the 181 cytotoxic isolates adhered to HEp-2 or Caco-2 cells. HEp-2 cells were invaded by 43.6%, and Caco-2 cells by 49.7%, of the 181 cytotoxic isolates. The invasion index on HEp-2 cells ranged from 1.9 x 10(-1) to 8.9 x 10(-6), whereas the invasion index on Caco-2 cells varied between 7.7 x 10(-2) and 8.3 x 10(-6). The most commonly isolated genera with

Kynetic resazurin assay (KRA) for bacterial quantification of foodborne pathogens

NASA Astrophysics Data System (ADS)

Arenas, Yaxal; Mandel, Arkady; Lilge, Lothar

2012-03-01

Fast detection of bacterial concentrations is important for the food industry and for healthcare. Early detection of infections and appropriate treatment is essential since, the delay of treatments for bacterial infections tends to be associated with higher mortality rates. In the food industry and in healthcare, standard procedures require the count of colony-forming units in order to quantify bacterial concentrations, however, this method is time consuming and reports require three days to be completed. An alternative is metabolic-colorimetric assays which provide time efficient in vitro bacterial concentrations. A colorimetric assay based on Resazurin was developed as a time kinetic assay (KRA) suitable for bacterial concentration measurements. An optimization was performed by finding excitation and emission wavelengths for fluorescent acquisition. A comparison of two non-related bacteria, foodborne pathogens Escherichia coli and Listeria monocytogenes, was performed in 96 well plates. A metabolic and clonogenic dependence was established for fluorescent kinetic signals.

Physical impaction injury effects on bacterial cells during spread plating influenced by cell characteristics of the organisms.

PubMed

Thomas, P; Mujawar, M M; Sekhar, A C; Upreti, R

2014-04-01

To understand the factors that contribute to the variations in colony-forming units (CFU) in different bacteria during spread plating. Employing a mix culture of vegetative cells of ten organisms varying in cell characteristics (Gram reaction, cell shape and cell size), spread plating to the extent of just drying the agar surface (50-60 s) was tested in comparison with the alternate spotting-and-tilt-spreading (SATS) approach where 100 μl inoculum was distributed by mere tilting of plate after spotting as 20-25 microdrops. The former imparted a significant reduction in CFU by 20% over the spreader-independent SATS approach. Extending the testing to single organisms, Gram-negative proteobacteria with relatively larger cells (Escherichia, Enterobacter, Agrobacterium, Ralstonia, Pantoea, Pseudomonas and Sphingomonas spp.) showed significant CFU reduction with spread plating except for slow-growing Methylobacterium sp., while those with small rods (Xenophilus sp.) and cocci (Acinetobacter sp.) were less affected. Among Gram-positive nonspore formers, Staphylococcus epidermidis showed significant CFU reduction while Staphylococcus haemolyticus and actinobacteria (Microbacterium, Cellulosimicrobium and Brachybacterium spp.) with small rods/cocci were unaffected. Vegetative cells of Bacillus pumilus and B. subtilis were generally unaffected while others with larger rods (B. thuringiensis, Brevibacillus, Lysinibacillus and Paenibacillus spp.) were significantly affected. A simulated plating study coupled with live-dead bacterial staining endorsed the chances of cell disruption with spreader impaction in afflicted organisms. Significant reduction in CFU could occur during spread plating due to physical impaction injury to bacterial cells depending on the spreader usage and the variable effects on different organisms are determined by Gram reaction, cell size and cell shape. The inoculum spreader could impart physical disruption of vegetative cells against a hard surface

Nonrecovery of varying proportions of viable bacteria during spread plating governed by the extent of spreader usage and proposal for an alternate spotting-spreading approach to maximize the CFU.

PubMed

Thomas, P; Sekhar, A C; Mujawar, M M

2012-08-01

To elucidate the cause of high variations and inconsistencies in bacterial CFU observed within and between different experiments while assessing viable bacterial counts through spread plating (SP). Following the inconsistent results, CFU estimations were undertaken through conventional SP using the spreader, or a modified approach that did not use spreader employing four organisms. The latter approach involving spotting-and-tilt-spreading of inoculum on agar surface [spotting spreading (SS)] yielded higher CFU by 11-120% over the weighted average depending on the organism and diluent. The adverse effect owing to the spreader was the most obvious in Escherichia coli followed by Staphylococcus epidermidis, Enterobacter cloacae and Bacillus pumilus. Plate attributes that determined the surface moisture levels of agar medium and the spreading practice adopted by the personnel formed two other major influencing factors. Plating for shorter periods (<60 s) using fresh 15/20 ml plates caused loss of 3-12% CFU owing to inoculum adhesion to spreader irrespective of glass or polypropylene make. On the other hand, prolonging the plating brought down the CFU significantly. Spreader movement on agar surface subsequent to the exhaustion of free moisture, which was marked by the experiencing of some friction to smooth spreader movement, was detrimental to vegetative cells, while Bacillus spores were less affected. The study brings out that the way SP is carried out exerts significant effects on CFU influenced by plate conditions. Prolonged use of spreader on dry agar surface could be highly detrimental to bacterial cells. A mild use of spreader accounting for spreader-adhering inoculum or the practice of SS not involving the spreader is recommended. This study unravels the effects owing to the spreader on bacterial cells and the CFU and recommends an alternate approach of SS to minimize CFU inconsistencies and to maximize the viable bacterial counts. © 2012 The Authors Journal

Long-Term Bacterial Dynamics in a Full-Scale Drinking Water Distribution System

PubMed Central

Prest, E. I.; Weissbrodt, D. G.; Hammes, F.; van Loosdrecht, M. C. M.; Vrouwenvelder, J. S.

2016-01-01

Large seasonal variations in microbial drinking water quality can occur in distribution networks, but are often not taken into account when evaluating results from short-term water sampling campaigns. Temporal dynamics in bacterial community characteristics were investigated during a two-year drinking water monitoring campaign in a full-scale distribution system operating without detectable disinfectant residual. A total of 368 water samples were collected on a biweekly basis at the water treatment plant (WTP) effluent and at one fixed location in the drinking water distribution network (NET). The samples were analysed for heterotrophic plate counts (HPC), Aeromonas plate counts, adenosine-tri-phosphate (ATP) concentrations, and flow cytometric (FCM) total and intact cell counts (TCC, ICC), water temperature, pH, conductivity, total organic carbon (TOC) and assimilable organic carbon (AOC). Multivariate analysis of the large dataset was performed to explore correlative trends between microbial and environmental parameters. The WTP effluent displayed considerable seasonal variations in TCC (from 90 × 103 cells mL-1 in winter time up to 455 × 103 cells mL-1 in summer time) and in bacterial ATP concentrations (<1–3.6 ng L-1), which were congruent with water temperature variations. These fluctuations were not detected with HPC and Aeromonas counts. The water in the network was predominantly influenced by the characteristics of the WTP effluent. The increase in ICC between the WTP effluent and the network sampling location was small (34 × 103 cells mL-1 on average) compared to seasonal fluctuations in ICC in the WTP effluent. Interestingly, the extent of bacterial growth in the NET was inversely correlated to AOC concentrations in the WTP effluent (Pearson’s correlation factor r = -0.35), and positively correlated with water temperature (r = 0.49). Collecting a large dataset at high frequency over a two year period enabled the characterization of previously

Long-Term Bacterial Dynamics in a Full-Scale Drinking Water Distribution System.

PubMed

Prest, E I; Weissbrodt, D G; Hammes, F; van Loosdrecht, M C M; Vrouwenvelder, J S

2016-01-01

Large seasonal variations in microbial drinking water quality can occur in distribution networks, but are often not taken into account when evaluating results from short-term water sampling campaigns. Temporal dynamics in bacterial community characteristics were investigated during a two-year drinking water monitoring campaign in a full-scale distribution system operating without detectable disinfectant residual. A total of 368 water samples were collected on a biweekly basis at the water treatment plant (WTP) effluent and at one fixed location in the drinking water distribution network (NET). The samples were analysed for heterotrophic plate counts (HPC), Aeromonas plate counts, adenosine-tri-phosphate (ATP) concentrations, and flow cytometric (FCM) total and intact cell counts (TCC, ICC), water temperature, pH, conductivity, total organic carbon (TOC) and assimilable organic carbon (AOC). Multivariate analysis of the large dataset was performed to explore correlative trends between microbial and environmental parameters. The WTP effluent displayed considerable seasonal variations in TCC (from 90 × 103 cells mL-1 in winter time up to 455 × 103 cells mL-1 in summer time) and in bacterial ATP concentrations (<1-3.6 ng L-1), which were congruent with water temperature variations. These fluctuations were not detected with HPC and Aeromonas counts. The water in the network was predominantly influenced by the characteristics of the WTP effluent. The increase in ICC between the WTP effluent and the network sampling location was small (34 × 103 cells mL-1 on average) compared to seasonal fluctuations in ICC in the WTP effluent. Interestingly, the extent of bacterial growth in the NET was inversely correlated to AOC concentrations in the WTP effluent (Pearson's correlation factor r = -0.35), and positively correlated with water temperature (r = 0.49). Collecting a large dataset at high frequency over a two year period enabled the characterization of previously

Rapid surface colony counts determination with three new miniaturised techniques.

PubMed

Malik, K A

1977-01-01

Loop-spreader form. From each bacterial suspension 10 micronl were carried and spread on each mini-disc. The method is useful for pathogenic organisms as the loop can readily be flame sterilized. For routine purposes where only approximate numbers of bacteria need to be known a still rapid semiquantitative method was deviced making use of a calibrated stainless steel Stamping-disc (Fig. 2c). A disc of 25mm diameter and 1 mm thickness delivered approximateyl 10 microlitres of supensions and was found to be most useful to stamp seven dilutions on a single plate. In collections and bacteriology laboratories where by conventional methods large number of plates are to be plated and counted the presented techniques could prove most convenient, rapid and economical.

Rapid and direct detection of Invivo kinetics of pathogenic bacterial infection from mouse blood and urine.

PubMed

Gopal, Judy; Lee, Chia-Hsun; Wu, Hui-Fen

2012-06-06

This study demonstrates the first use of matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS) to trace the Invivo infection kinetics of the well known deadly pathogen Staphylococcus aureus in Swiss albino mice. The growth curve of the bacteria from the point of injection (200μL of bacterial suspension (10(8)cfu/mL)) into the mouse blood till mortality (death) was periodically analyzed using the plate counting method and MALDI-MS. Bacterial counts of 10(3)cfu/mL were observed in the log phase of the growth curve in the blood and 10(2)cfu/mL were observed in the urine samples. Death occurred in the log phase of the growth curve, where the bacterial counts showed steady increase. In other cases, the bacteria counts started decreasing after 48h and by 96h the bacteria got totally eliminated from the mouse and these mice survived. Direct MALDI-MS was not feasible for tracking the bacteria in the infected blood. However, ionic liquid 1-Butyl-3-methylimidazolium tetrafluoroborate was successful in enabling bacterial detection amidst the strong blood peaks. But, in the case of the urine analysis, it was observed that direct MALDI-MS was adequate to enable detection. The results obtained prove the efficacy of MALDI-MS for analyzing pathogenic bacteria in clinical samples. This article is part of a Special Issue entitled: Proteomics: The clinical link. Copyright © 2011 Elsevier B.V. All rights reserved.

Bacteria killing nanotechnology Bio-Kil effectively reduces bacterial burden in intensive care units.

PubMed

Hsueh, P-R; Huang, H-C; Young, T-G; Su, C-Y; Liu, C-S; Yen, M-Y

2014-04-01

A contaminated hospital environment has been identified as an important reservoir of pathogens causing healthcare-associated infections. This study is to evaluate the efficacy of bacteria killing nanotechnology Bio-Kil on reducing bacterial counts in an intensive care unit (ICU). Two single-bed rooms (S-19 and S-20) in the ICU were selected from 7 April to 27 May 2011. Ten sets of new textiles (pillow cases, bed sheets, duvet cover, and patient clothing) used by patients in the two single-bed rooms were provided by the sponsors. In the room S-20, the 10 sets of new textiles were washed with Bio-Kil; the room walls, ceiling, and air-conditioning filters were treated with Bio-Kil; and the surfaces of instruments (respirator, telephone, and computer) were covered with Bio-Kil-embedded silicon pads. Room S-19 served as the control. We compared the bacterial count on textiles and environment surfaces as well as air samples between the two rooms. A total of 1,364 samples from 22 different sites in each room were collected. The mean bacterial count on textiles and environmental surfaces in room S-20 was significantly lower than that in room S-19 (10.4 vs 49.6 colony-forming units [CFU]/100 cm(2); P < 0.001). Room S-20 had lower bacterial counts in air samples than room S-19 (33.4-37.6 vs 21.6-25.7 CFU/hour/plate; P < 0.001). The density of microbial isolations was significantly greater among patients admitted to room S-19 than those to room S-20 (9.15 vs 5.88 isolates per 100 patient-days, P < 0.05). Bio-Kil can significantly reduce bacterial burden in the environment of the ICU.

The Immediate and Delayed Post-Debridement Effects on Tissue Bacterial Wound Counts of Hypochlorous Acid Versus Saline Irrigation in Chronic Wounds.

PubMed

Hiebert, John M; Robson, Martin C

2016-01-01

Introduction: Wound debridement is considered essential in chronic wound management. Hypochlorous acid has been shown to be an effective agent in reducing wound bacterial counts in open wounds. Ultrasound-enabled wound debridement is an effective and efficient method of debridement. This study compared ultrasound irrigation with hypochlorous acid versus saline irrigation for wound debridement on pre- and postoperative wounds and determined regrowth of bacteria over 1 week period of time. Finally, the outcome of definitive wound closure of the clinically clean-appearing wounds was recorded. Methods: Seventeen consenting adult patients with chronic open wounds were randomly selected for study. The patients were randomly divided into the hypochlorous acid irrigation or saline irrigation group. All patients provided pre- and postoperative tissue samples for qualitative and quantitative bacteriology. For the time (7 days) between the debridement procedure and the definitive closure procedure, the wounds were dressed with a silver-impregnated dressing and a hydroconductive dressing. Results : Both types of irrigation in the ultrasonic system initially lowered the bacterial counts by 4 to 6 logs. However, by the time of definitive closure, the saline-irrigated wounds had bacterial counts back up to 10 5 whereas the hypochlorous acid-irrigated wounds remained at 10 2 or fewer. More than 80% of patients in the saline group had postoperative closure failure compared with 25% of patients in the hypochlorous acid group. Conclusions: Hypochlorous acid irrigation with ultrasound debridement reduced bacterial growth in chronic open wounds more efficiently than saline alone. Postoperative wound closure outcomes suggest a remarkable reduction in wound complications after wound debridement using hypochlorous acid irrigation with ultrasound versus saline alone.

The effect of an antibacterial washing-up liquid in reducing dishwater aerobic plate counts.

PubMed

Holah, J T; Hall, K E

2006-05-01

To assess any significant differences in the aerobic plate count (APC) of catering dishwaters following the use of a traditional, nonantibacterial or an antibacterial washing-up liquid. A dishwashing trial was undertaken within a commercial restaurant of 6 weeks duration (3 weeks with each washing-up liquid in a randomized, weekly pattern). Five replicate samples were taken from the dishwater at the end of the washing-up operation, on three separate occasions each day corresponding to mid-morning, lunchtime and mid-afternoon meal preparations. The antibacterial product was shown to significantly reduce the APC by an average log10 reduction of 1.81 CFU ml(-1) (98.5%) as compared with the traditional product. APC were lower for each of the three weekly time periods for the antibacterial product. Continued use of the antibacterial product did not decrease the APC of the dishwater, though with the traditional product, dishwater counts increased throughout the trial week. Antibacterial washing-up liquids, with proven activity in controlling levels of microorganisms in dishwaters, could play a significant role in reducing the risk of cross-contamination between washed articles during washing-up operations.

The degree of bacterial contamination while performing spine surgery.

PubMed

Ahn, Dong Ki; Park, Hoon Seok; Kim, Tae Woo; Yang, Jong Hwa; Boo, Kyung Hwan; Kim, In Ja; Lee, Hye Jin

2013-03-01

Prospective experimental study. To evaluate bacterial contamination during surgery. The participants of surgery and ventilation system have been known as the most significant sources of contamination. Two pairs of air culture blood agar plate for G(+) bacteria and MacConkey agar plate for G(-) bacteria were placed at 3 different locations in a conventional operation room: in the surgical field, under the airflow of local air conditioner, and pathway to door while performing spine surgeries. One pair of culture plates was retrieved after one hour and the other pair was retrieved after 3 hours. The cultured bacteria were identified and number of colonies was counted. There was no G(-) bacteria identified. G(+) bacteria grew on all 90 air culture blood agar plates. The colony count of one hour group was 14.5±5.4 in the surgical field, 11.3±6.6 under the local air conditioner, and 13.1±8.7 at the pathway to the door. There was no difference among the 3 locations. The colony count of 3 hours group was 46.4±19.5, 30.3±12.9, and 39.7±15.2, respectively. It was more at the surgical field than under the air conditioner (p=0.03). The number of colonies of one hour group was 13.0±7.0 and 3 hours group was 38.8±17.1. There was positive correlation between the time and the number of colonies (r=0.76, p=0.000). Conventional operation room was contaminated by G(+) bacteria. The degree of contamination was most high at the surgical field. The number of bacteria increased right proportionally to the time.

The Degree of Bacterial Contamination While Performing Spine Surgery

PubMed Central

Ahn, Dong Ki; Park, Hoon Seok; Yang, Jong Hwa; Boo, Kyung Hwan; Kim, In Ja; Lee, Hye Jin

2013-01-01

Study Design Prospective experimental study. Purpose To evaluate bacterial contamination during surgery. Overview of Literature The participants of surgery and ventilation system have been known as the most significant sources of contamination. Methods Two pairs of air culture blood agar plate for G(+) bacteria and MacConkey agar plate for G(-) bacteria were placed at 3 different locations in a conventional operation room: in the surgical field, under the airflow of local air conditioner, and pathway to door while performing spine surgeries. One pair of culture plates was retrieved after one hour and the other pair was retrieved after 3 hours. The cultured bacteria were identified and number of colonies was counted. Results There was no G(-) bacteria identified. G(+) bacteria grew on all 90 air culture blood agar plates. The colony count of one hour group was 14.5±5.4 in the surgical field, 11.3±6.6 under the local air conditioner, and 13.1±8.7 at the pathway to the door. There was no difference among the 3 locations. The colony count of 3 hours group was 46.4±19.5, 30.3±12.9, and 39.7±15.2, respectively. It was more at the surgical field than under the air conditioner (p=0.03). The number of colonies of one hour group was 13.0±7.0 and 3 hours group was 38.8±17.1. There was positive correlation between the time and the number of colonies (r=0.76, p=0.000). Conclusions Conventional operation room was contaminated by G(+) bacteria. The degree of contamination was most high at the surgical field. The number of bacteria increased right proportionally to the time. PMID:23508998

Accurate live and dead bacterial cell enumeration using flow cytometry (Conference Presentation)

NASA Astrophysics Data System (ADS)

Ou, Fang; McGoverin, Cushla; Swift, Simon; Vanholsbeeck, Frédérique

2017-03-01

Flow cytometry (FCM) is based on the detection of scattered light and fluorescence to identify cells with particular characteristics of interest. However most FCM cannot precisely control the flow through its interrogation point and hence the volume and concentration of the sample cannot be immediately obtained. The easiest, most reliable and inexpensive way of obtaining absolute counts with FCM is by using reference beads. We investigated a method of using FCM with reference beads to measure live and dead bacterial concentration over the range of 106 to 108 cells/mL and ratio varying from 0 to 100%. We believe we are the first to use this method for such a large cell concentration range while also establishing the effect of varying the live/dead bacteria ratios. Escherichia coli solutions with differing ratios of live:dead cells were stained with fluorescent dyes SYTO 9 and propidium iodide (PI), which label live and dead cells, respectively. Samples were measured using a LSR II Flow Cytometer (BD Biosciences); using 488 nm excitation with 20 mW power. Both SYTO 9 and PI fluorescence were collected and threshold was set to side scatter. Traditional culture-based plate count was done in parallel to the FCM analysis. The concentration of live bacteria from FCM was compared to that obtained by plate counts. Preliminary results show that the concentration of live bacteria obtained by FCM and plate counts correlate well with each other and indicates this may be extended to a wider concentration range or for studying other cell characteristics.

Infection control in digital intraoral radiography: evaluation of microbiological contamination of photostimulable phosphor plates in barrier envelopes.

PubMed

MacDonald, David S; Waterfield, J Douglas

2011-01-01

The detectors (both solid-state sensors and photostimulable phosphor [PSP] plates) used for digital intraoral radiography cannot be autoclaved, and barriers are typically used to prevent the spread of infection. The aim of this study was to determine the effectiveness of a barrier envelope system for PSP plates. Disinfected PSP plates were aseptically inserted into barrier envelopes and placed in a periapical location. One PSP plate was placed in each of 28 patients, and 12 plates in each of 2 volunteers (D.S.M., J.D.W.). After retrieval, each PSP plate was removed from its barrier envelope, immersed in trypticase soy broth and aliquots were plated on trypticase soy agar. Bacterial colonies were counted 2 days later. Fifty-two PSP plates in barrier envelopes were evaluated for contamination. Quality assurance of the PSP plates before clinical placement revealed defects in the integrity of 4 barrier envelopes, caused by forceps-related damage or failure to achieve a uniform seal. These defects allowed substantial contamination. Contamination also occurred as a result of failure to extract the PSP plate from the barrier envelope cleanly. Of the 44 barriers with no obvious defects that were placed by either final-year dental students or a radiologist, only 3 allowed bacterial contamination of the PSP plate. Detectors contained in barrier envelopes remain a potential source of contamination. PSP plates must be disinfected between removal from a contaminated barrier envelope and placement in a new barrier envelope. In addition, placement into the barrier envelope should ideally be carried out under aseptic conditions. Finally, the integrity of each sealed barrier envelope must be verified visually before release to the clinic.

Evaluation of the limulus amoebocyte lysate test in conjunction with a gram negative bacterial plate count for detecting irradiation of chicken

NASA Astrophysics Data System (ADS)

Scotter, Susan L.; Wood, Roger; McWeeny, David J.

A study to evaluate the potential of the Limulus amoebocyte lysate (LAL) test in conjuction with a Gram negative bacteria (GNB) plate count for detecting the irradiation of chicken is described. Preliminary studies demonstrated that chickens irradiated at an absorbed dose of 2.5 kGy could be differentiated from unirradiated birds by measuring levels of endotoxin and of numbers of GNB on chicken skin. Irradiated birds were found to have endotoxin levels similar to those found in unirradiated birds but significantly lower numbers of GNB. In a limited study the test was found to be applicable to birds from different processors. The effect of temperature abuse on the microbiological profile, and thus the efficacy of the test, was also investigated. After temperature abuse, the irradiated birds were identifiable at worst up to 3 days after irradiation treatment at the 2.5 kGy level and at best some 13 days after irradiation. Temperature abuse at 15°C resulted in rapid recovery of surviving micro-organisms which made differentiation of irradiated and unirradiated birds using this test unreliable. The microbiological quality of the bird prior to irradiation treatment also affected the test as large numbers of GNB present on the bird prior to irradiation treatment resulted in larger numbers of survivors. In addition, monitoring the developing flora after irradiation treatment and during subsequent chilled storage also aided differentiation of irradiated and unirradiated birds. Large numbers of yeasts and Gram positive cocci were isolated from irradiated carcasses whereas Gram negative oxidative rods were the predominant spoilage flora on unirradiated birds.

Evaluation of the effects of footwear hygiene protocols on nonspecific bacterial contamination of floor surfaces in an equine hospital.

PubMed

Stockton, Kelly A; Morley, Paul S; Hyatt, Doreene R; Burgess, Brandy A; Patterson, Gage; Dunowska, Magda; Lee, David E

2006-04-01

To evaluate the effects of footwear hygiene protocols on bacterial contamination of floor surfaces in an equine hospital. Field trial. Footwear hygiene protocols evaluated included use of rubber overboots with footbaths and footmats containing a quaternary ammonium disinfectant, rubber overboots with footbaths and footmats containing a peroxygen disinfectant, and no restrictions on footwear type but mandatory use of footbaths and footmats containing a peroxygen disinfectant. Nonspecific aerobic bacterial counts were determined via 2 procedures for sample collection and bacterial enumeration (contact plates vs swabbing combined with use of spread plates), and the effects of each footwear hygiene protocol were compared. There were no consistent findings suggesting that any of the protocols were associated with differences in numbers of bacteria recovered from floor surfaces. Although there were detectable differences in numbers of bacteria recovered in association with different footwear hygiene protocols, differences in least square mean bacterial counts did not appear to be clinically relevant (ie, were < 1 log10). Although cleaning and disinfection of footwear are important aids in reducing the risk of nosocomial transmission of infectious agents in veterinary hospitals, the numbers of aerobic bacteria recovered from floor surfaces were not affected by use of rubber overboots or the types of disinfectant used in this study. Further study is warranted to evaluate the usefulness of footwear hygiene practices relative to their efficacy for reducing transmission of specific pathogens or decreasing nosocomial disease risk.

Hydrophilic-treated plastic plates for wide-range analysis of Giemsa-stained red blood cells and automated Plasmodium infection rate counting.

PubMed

Hashimoto, Muneaki; Yatsushiro, Shouki; Yamamura, Shohei; Tanaka, Masato; Sakamoto, Hirokazu; Ido, Yusuke; Kajimoto, Kazuaki; Bando, Mika; Kido, Jun-Ichi; Kataoka, Masatoshi

2017-08-08

Malaria is a red blood cell (RBC) infection caused by Plasmodium parasites. To determine RBC infection rate, which is essential for malaria study and diagnosis, microscopic evaluation of Giemsa-stained thin blood smears on glass slides ('Giemsa microscopy') has been performed as the accepted gold standard for over 100 years. However, only a small area of the blood smear provides a monolayer of RBCs suitable for determination of infection rate, which is one of the major reasons for the low parasite detection rate by Giemsa microscopy. In addition, because Giemsa microscopy is exacting and time-consuming, automated counting of infection rates is highly desirable. A method that allows for microscopic examination of Giemsa-stained cells spread in a monolayer on almost the whole surface of hydrophilic-treated cyclic olefin copolymer (COC) plates was established. Because wide-range Giemsa microscopy can be performed on a hydrophilic-treated plate, the method may enable more reliable diagnosis of malaria in patients with low parasitaemia burden. Furthermore, the number of RBCs and parasites stained with a fluorescent nuclear staining dye could be counted automatically with a software tool, without Giemsa staining. As a result, researchers studying malaria may calculate the infection rate easily, rapidly, and accurately even in low parasitaemia. Because the running cost of these methods is very low and they do not involve complicated techniques, the use of hydrophilic COC plates may contribute to improved and more accurate diagnosis and research of malaria.

Evaluation of quantitative PCR measurement of bacterial colonization of epithelial cells.

PubMed

Schmidt, Marcin T; Olejnik-Schmidt, Agnieszka K; Myszka, Kamila; Borkowska, Monika; Grajek, Włodzimierz

2010-01-01

Microbial colonization is an important step in establishing pathogenic or probiotic relations to host cells and in biofilm formation on industrial or medical devices. The aim of this work was to verify the applicability of quantitative PCR (Real-Time PCR) to measure bacterial colonization of epithelial cells. Salmonella enterica and Caco-2 intestinal epithelial cell line was used as a model. To verify sensitivity of the assay a competition of the pathogen cells to probiotic microorganism was tested. The qPCR method was compared to plate count and radiolabel approach, which are well established techniques in this area of research. The three methods returned similar results. The best quantification accuracy had radiolabel method, followed by qPCR. The plate count results showed coefficient of variation two-times higher than this of qPCR. The quantitative PCR proved to be a reliable method for enumeration of microbes in colonization assay. It has several advantages that make it very useful in case of analyzing mixed populations, where several different species or even strains can be monitored at the same time.

Flow cytometry analysis using sysmex UF-1000i classifies uropathogens based on bacterial, leukocyte, and erythrocyte counts in urine specimens among patients with urinary tract infections.

PubMed

Monsen, Tor; Rydén, Patrik

2015-02-01

Urinary tract infections (UTIs) are the second most common bacterial infection. Urine culture is the gold standard for diagnosis, but new techniques, such as flow cytometry analysis (FCA), have been introduced. The aim of the present study was to evaluate FCA characteristics regarding bacteriuria, leukocyturia, and erythrocyturia in relation to cultured uropathogens in specimens from patients with a suspected UTI. We also wanted to evaluate whether the FCA characteristics can identify uropathogens prior to culture. From a prospective study, 1,587 consecutive urine specimens underwent FCA prior to culture during January and February 2012. Outpatients and inpatients (79.6% and 19.4%, respectively) were included, of whom women represented 67.5%. In total, 620 specimens yielded growth, of which Escherichia coli represented 65%, Enterococcus spp. 8%, Klebsiella spp. 7%, and Staphylococcus spp. 5%. For the uropathogens, the outcome of FCA was compared against the results for specimens with E. coli and those with a negative culture. E. coli had high bacterial (median, 17,914/μl), leukocyte (median, 348/μl), and erythrocyte (median, 23/μl) counts. With the exception of Klebsiella spp., the majority of the uropathogens had considerable or significantly lower bacterial counts than that of E. coli. High leukocyte counts were found in specimens with Staphylococcus aureus, Proteus mirabilis, Pseudomonas aeruginosa, and group C streptococci. Elevated erythrocyte counts were found for P. vulgaris, P. aeruginosa, and group C streptococci, as well as for Staphylococcus saprophyticus. In essence, FCA adds new information about the bacterial, leukocyte, and erythrocyte counts in urine specimens for different uropathogens. Based on FCA characteristics, uropathogens can be classified and identified prior to culture. E. coli and Klebsiella spp. have similar FCA characteristics. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Fortifying fresh human milk with commercial powdered human milk fortifiers does not affect bacterial growth during 6 hours at room temperature.

PubMed

Telang, Sucheta; Berseth, Carol Lynn; Ferguson, Paul W; Kinder, Julie M; DeRoin, Mark; Petschow, Bryon W

2005-10-01

To evaluate the growth of resident aerobic mesophilic flora and added Enterobacter sakazakii in fresh, unfortified human milk; fresh human milk fortified with two commercial powdered fortifiers differing in iron content; and infant formula prepared from powder. Eight mothers provided preterm breast milk samples. Breast milk samples were divided into three aliquots: unfortified, fortified with fortifier containing 1.44 mg iron/14 kcal, and fortified with fortifier containing 0.4 mg iron/14 kcal. Aliquots of formula were prepared. Breast milk and formula aliquots were divided into two test samples. Half were inoculated with low amounts of E sakazakii; half were not. All test samples were maintained at room temperature (22 degrees C), serially diluted, and plated onto agars after 0, 2, 4, and 6 hours. Plates were incubated at 35 degrees C and enumerated. Data were analyzed using repeated measures analysis of variance. P<.05 was considered significant. There were no differences in colony counts of aerobic bacteria among uninoculated or among inoculated human milk samples at any time; counts did not increase significantly over 6 hours. There were no differences in colony counts of E sakazakii among inoculated human milk samples at any time; counts did not increase significantly over 6 hours. Aerobic bacteria and E sakazakii colony counts from infant formula did not increase significantly over 6 hours. During 6 hours at 22 degrees C, fresh human milk and formula had negligible bacterial growth; fortifying human milk with powdered fortifiers did not affect bacterial growth.

Detecting swift fox: Smoked-plate scent stations versus spotlighting

Treesearch

Daniel W. Uresk; Kieth E. Severson; Jody Javersak

2003-01-01

We compared two methods of detecting presence of swift fox: smoked-plate scent stations and spotlight counts. Tracks were counted on ten 1-mile (1.6-km) transects with bait/tracking plate stations every 0.1 mile (0.16 km). Vehicle spotlight counts were conducted on the same transects. Methods were compared with Spearman's rank order correlation. Repeated measures...

Strain Library Imaging Protocol for high-throughput, automated single-cell microscopy of large bacterial collections arrayed on multiwell plates.

PubMed

Shi, Handuo; Colavin, Alexandre; Lee, Timothy K; Huang, Kerwyn Casey

2017-02-01

Single-cell microscopy is a powerful tool for studying gene functions using strain libraries, but it suffers from throughput limitations. Here we describe the Strain Library Imaging Protocol (SLIP), which is a high-throughput, automated microscopy workflow for large strain collections that requires minimal user involvement. SLIP involves transferring arrayed bacterial cultures from multiwell plates onto large agar pads using inexpensive replicator pins and automatically imaging the resulting single cells. The acquired images are subsequently reviewed and analyzed by custom MATLAB scripts that segment single-cell contours and extract quantitative metrics. SLIP yields rich data sets on cell morphology and gene expression that illustrate the function of certain genes and the connections among strains in a library. For a library arrayed on 96-well plates, image acquisition can be completed within 4 min per plate.

A comparison of methods to assess the antimicrobial activity of nanoparticle combinations on bacterial cells.

PubMed

Bankier, Claire; Cheong, Yuen; Mahalingam, Suntharavathanan; Edirisinghe, Mohan; Ren, Guogang; Cloutman-Green, Elaine; Ciric, Lena

2018-01-01

Bacterial cell quantification after exposure to antimicrobial compounds varies widely throughout industry and healthcare. Numerous methods are employed to quantify these antimicrobial effects. With increasing demand for new preventative methods for disease control, we aimed to compare and assess common analytical methods used to determine antimicrobial effects of novel nanoparticle combinations on two different pathogens. Plate counts of total viable cells, flow cytometry (LIVE/DEAD BacLight viability assay) and qPCR (viability qPCR) were used to assess the antimicrobial activity of engineered nanoparticle combinations (NPCs) on Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacteria at different concentrations (0.05, 0.10 and 0.25 w/v%). Results were analysed using linear models to assess the effectiveness of different treatments. Strong antimicrobial effects of the three NPCs (AMNP0-2) on both pathogens could be quantified using the plate count method and flow cytometry. The plate count method showed a high log reduction (>8-log) for bacteria exposed to high NPC concentrations. We found similar antimicrobial results using the flow cytometry live/dead assay. Viability qPCR analysis of antimicrobial activity could not be quantified due to interference of NPCs with qPCR amplification. Flow cytometry was determined to be the best method to measure antimicrobial activity of the novel NPCs due to high-throughput, rapid and quantifiable results.

Pathogenic features of heterotrophic plate count bacteria from drinking-water boreholes.

PubMed

Horn, Suranie; Pieters, Rialet; Bezuidenhout, Carlos

2016-12-01

Evidence suggests that heterotrophic plate count (HPC) bacteria may be hazardous to humans with weakened health. We investigated the pathogenic potential of HPC bacteria from untreated borehole water, consumed by humans, for: their haemolytic properties, the production of extracellular enzymes such as DNase, proteinase, lipase, lecithinase, hyaluronidase and chondroitinase, the effect simulated gastric fluid has on their survival, as well as the bacteria's antibiotic-susceptible profile. HuTu-80 cells acted as model for the human intestine and were exposed to the HPC isolates to determine their effects on the viability of the cells. Several HPC isolates were α- or β-haemolytic, produced two or more extracellular enzymes, survived the SGF treatment, and showed resistance against selected antibiotics. The isolates were also harmful to the human intestinal cells to varying degrees. A novel pathogen score was calculated for each isolate. Bacillus cereus had the highest pathogen index: the pathogenicity of the other bacteria declined as follows: Aeromonas taiwanensis > Aeromonas hydrophila > Bacillus thuringiensis > Alcaligenes faecalis > Pseudomonas sp. > Bacillus pumilus > Brevibacillus sp. > Bacillus subtilis > Bacillus sp. These results demonstrated that the prevailing standards for HPCs in drinking water may expose humans with compromised immune systems to undue risk.

Site-specific mouth rinsing can improve oral odor by altering bacterial counts. Blind crossover clinical study.

PubMed

Alqumber, Mohammed A; Arafa, Khaled A

2014-11-01

To determine whether site-specific mouth rinsing with oral disinfectants can improve oral odor beyond the traditional panoral mouth disinfection with mouth rinses by targeting specifically oral malodor implicated anaerobic bacteria. Twenty healthy fasting subjects volunteered for a blinded prospective, descriptive correlational crossover cross-section clinical trial conducted during the month of Ramadan between July and August 2013 in Albaha province in Saudi Arabia involving the application of Listerine Cool Mint mouth rinse by either the traditional panoral rinsing method, or a site-specific disinfection method targeting the subgingival and supragingival plaque and the posterior third of the tongue dorsum, while avoiding the remaining locations within the oral cavity. The viable anaerobic and aerobic bacterial counts, volatile sulfur compounds (VSCs) levels, organoleptic assessment of oral odor, and the tongue-coating index were compared at baseline, one, 5, and 9 hours after the treatment. The site-specific disinfection method reduced the VSCs and anaerobic bacterial loads while keeping the aerobic bacterial numbers higher than the traditional panoral rinsing method. Site-specific disinfection can more effectively maintain a healthy oral cavity by predominantly disinfecting the niches of anaerobic bacteria within the oral cavity.

Effect of species, breed, and age on bacterial load in bovine and bubaline semen

PubMed Central

Sannat, Chandrahas; Nair, Ajit; Sahu, S. B.; Sahasrabudhe, S. A.; Kumar, Ashish; Gupta, Amit Kumar; Shende, R. K.

2015-01-01

Aim: The present study was conducted to investigate the effect of species, breed and age on bacterial load in fresh and frozen semen of Cattle and Buffalo bull. Materials and Methods: Present study covered 56 cow and 10 buffalo bulls stationed at Central Semen Station Anjora, Durg (Chhattisgarh). Impact of breeds on bacterial load in semen was assessed using six breeds of cattle viz. Sahiwal, Gir, Red Sindhi, Tharparkar, Jersey and Holstein Friesian (HF) cross. Cow bulls were categorized into four different groups based on their age (<4 years, 4-5 years, 5-6 years and > 6 years) to study variation among age groups. Bacterial load was measured in fresh and frozen semen samples from these bulls using the standard plate count (SPC) method and count was expressed as colony forming unit (CFU) per ml of semen. Results: Higher bacterial load was reported in fresh (2.36 × 104 ± 1943 CFU/ml) and frozen (1.00 × 10 ± 90 CFU/ml) semen of cow bulls as compared to buffalo bulls (1.95 × 104 ± 2882 and 7.75 × 102 ± 160 CFU/ml in fresh and frozen semen, respectively). Jersey bull showed significantly higher bacterial count (p < 0.05) both in fresh (4.07 × 104 ± 13927 CFU/ml) and frozen (1.92 × 103 ± 178 CFU/ml) semen followed by HF cross, Sahiwal, Gir, Red Sindhi and Tharparkar bull. Bulls aged < 4 years and more than 6 years yielded increased bacterial load in their semen. Although a minor variation was reported between species and among age groups, no significant differences were measured. Conclusion: Bacterial load in semen did not differ significantly between species and age groups; however significant variation was reported among different breeds. Bulls of Jersey breed showed significantly higher bacterial load in semen as compared to the crossbred and indigenous bull. PMID:27047115

Effectiveness of Polyvalent Bacterial Lysate and Autovaccines Against Upper Respiratory Tract Bacterial Colonization by Potential Pathogens: A Randomized Study

PubMed Central

Zagólski, Olaf; Stręk, Paweł; Kasprowicz, Andrzej; Białecka, Anna

2015-01-01

Background Polyvalent bacterial lysate (PBL) is an oral immunostimulating vaccine consisting of bacterial standardized lysates obtained by lysis of different strains of bacteria. Autovaccines are individually prepared based on the results of smears obtained from the patient. Both types of vaccine can be used to treat an ongoing chronic infection. This study sought to determine which method is more effective against nasal colonization by potential respiratory tract pathogens. Material/Methods We enrolled 150 patients with aerobic Gram stain culture and count results indicating bacterial colonization of the nose and/or throat by potential pathogens. The participants were randomly assigned to each of the following groups: 1. administration of PBL, 2. administration of autovaccine, and 3. no intervention (controls). Results Reduction of the bacterial count in Streptococcus pneumoniae-colonized participants was significant after the autovaccine (p<0.001) and PBL (p<0.01). Reduction of the bacterial count of other β-hemolytic streptococcal strains after treatment with the autovaccine was significant (p<0.01) and was non-significant after PBL. In Haemophilus influenzae colonization, significant reduction in the bacterial count was noted in the PBL group (p<0.01). Methicillin-resistant Staphylococcus aureus colonization did not respond to either treatment. Conclusions The autovaccine is more effective than PBL for reducing bacterial count of Streptococcus pneumoniae and β-hemolytic streptococci, while PBL was more effective against Haemophilus influenzae colonization. PMID:26434686

On-farm and postharvest processing sources of bacterial contamination to melon rinds.

PubMed

Gagliardi, J V; Millner, P D; Lester, G; Ingram, D

2003-01-01

Multistate and international foodborne illness outbreaks, particularly involving cantaloupe and often involving rare Salmonella spp., have increased dramatically over the past 13 years. This study assessed the sources and extent of melon rind contamination in production fields and at processing and packing facilities. In the spring of 1999, cantaloupe (Cucumis melo L. [reticulatus group] cv. Cruiser) sampled from two sites in the Rio Grande River Valley showed that postharvest-processed melon rinds often had greater plate counts of bacterial contaminants than field-fresh melons. Cantaloupe in the field had 2.5 to 3.5 log CFU g(-1) rind total coliforms by aerobic plate counts, whereas washed melons had 4.0 to 5.0 log CFU g(-1). In the fall of 1999, coliforms on honeydew melons (C. melo [inodorous group] cv. Honey Brew) ranged from 2.6 to 3.7 log CFU g(-1) after processing, and total and fecal coliforms and enterococci never fell below 2.5 log CFU g(-1). A hydrocooler at another site contaminated cantaloupe rinds with up to 3.4 log CFU g(-1) total and fecal enterococci; a secondary rinse with chlorinated water incompletely removed these bacteria. Sources of coliforms and enterococci were at high levels in melon production soils, especially in furrows that were flood irrigated, in standing water at one field, and in irrigation water at both sites. At one processing facility, wash water pumped from the Rio Grande River may not have been sufficiently disinfected prior to use. Because soil, irrigation water, and process water were potential sources of bacterial contamination, monitoring and management on-farm and at processing and packing facilities should focus on water quality as an important control point for growers and packers to reduce bacterial contamination on melon rinds.

Comparison of fluorescence microscopy and solid-phase cytometry methods for counting bacteria in water

USGS Publications Warehouse

Lisle, John T.; Hamilton, Martin A.; Willse, Alan R.; McFeters, Gordon A.

2004-01-01

Total direct counts of bacterial abundance are central in assessing the biomass and bacteriological quality of water in ecological and industrial applications. Several factors have been identified that contribute to the variability in bacterial abundance counts when using fluorescent microscopy, the most significant of which is retaining an adequate number of cells per filter to ensure an acceptable level of statistical confidence in the resulting data. Previous studies that have assessed the components of total-direct-count methods that contribute to this variance have attempted to maintain a bacterial cell abundance value per filter of approximately 106 cells filter-1. In this study we have established the lower limit for the number of bacterial cells per filter at which the statistical reliability of the abundance estimate is no longer acceptable. Our results indicate that when the numbers of bacterial cells per filter were progressively reduced below 105, the microscopic methods increasingly overestimated the true bacterial abundance (range, 15.0 to 99.3%). The solid-phase cytometer only slightly overestimated the true bacterial abundances and was more consistent over the same range of bacterial abundances per filter (range, 8.9 to 12.5%). The solid-phase cytometer method for conducting total direct counts of bacteria was less biased and performed significantly better than any of the microscope methods. It was also found that microscopic count data from counting 5 fields on three separate filters were statistically equivalent to data from counting 20 fields on a single filter.

High level bacterial contamination of secondary school students' mobile phones.

PubMed

Kõljalg, Siiri; Mändar, Rando; Sõber, Tiina; Rööp, Tiiu; Mändar, Reet

2017-06-01

While contamination of mobile phones in the hospital has been found to be common in several studies, little information about bacterial abundance on phones used in the community is available. Our aim was to quantitatively determine the bacterial contamination of secondary school students' mobile phones. Altogether 27 mobile phones were studied. The contact plate method and microbial identification using MALDI-TOF mass spectrometer were used for culture studies. Quantitative PCR reaction for detection of universal 16S rRNA, Enterococcus faecalis 16S rRNA and Escherichia coli allantoin permease were performed, and the presence of tetracycline ( tet A, tet B, tet M), erythromycin ( erm B) and sulphonamide ( sul 1) resistance genes was assessed. We found a high median bacterial count on secondary school students' mobile phones (10.5 CFU/cm 2 ) and a median of 17,032 bacterial 16S rRNA gene copies per phone. Potentially pathogenic microbes ( Staphylococcus aureus , Acinetobacter spp. , Pseudomonas spp., Bacillus cereus and Neisseria flavescens ) were found among dominant microbes more often on phones with higher percentage of E. faecalis in total bacterial 16S rRNA. No differences in contamination level or dominating bacterial species between phone owner's gender and between phone types (touch screen/keypad) were found. No antibiotic resistance genes were detected on mobile phone surfaces. Quantitative study methods revealed high level bacterial contamination of secondary school students' mobile phones.

Effect of Probiotic Lactobacillus reuteri on Salivary Cariogenic Bacterial Counts among Groups of Preschool Children in Jeddah, Saudi Arabia: A Randomized Clinical Trial.

PubMed

Alamoudi, Najlaa M; Almabadi, Eman S; El Ashiry, Eman A; El Derwi, Douaa A

2018-05-15

To evaluate the effect of probiotic Lactobacilli reuteri lozenges on caries-associated salivary bacterial counts (Mutans streptococci and Lactobacillus), dental plaque accumulation, and salivary buffer capacity in a group of preschool children. The study group consisted of 178 healthy children (aged 3-6 years). Children were randomly grouped: the experimental group (n = 90) received L. reuteri probiotic lozenges and the control group (n = 88) received placebo lozenges, twice daily, for 28 days. Salivary Mutans streptococci and Lactobacillus counts, and buffer capacity were assessed using chair-side caries-risk test (CRT®) kits. The Simplified Oral Hygiene index (OHI-S) was used to assess dental plaque accumulation at baseline and after 28 days. After 28 days, the experimental group had a statistically significant reduction in Mutans streptococci and lactobacilli (p = 0.000 and p = 0.020, respectively) and both groups had less plaque accumulation than at baseline. While the buffer capacity in the experimental group increased more than in the control group, it was not statistically significant (p = 0.577). Compliance was 90%, with no adverse events. Consumption of probiotic lozenges containing L. reuteri reduces caries-associated bacterial counts significantly. Probiotics consumption may have a beneficial caries-preventive effect.

Bacterial, Fungal, and Actinomycete Populations in Soils Receiving Repeated Applications of 2,4-Dichlorophenoxyacetic Acid and Trifluralin 1

PubMed Central

Breazeale, F. W.; Camper, N. D.

1970-01-01

Soil samples were collected from an untreated plot and plots receiving repeated applications of 2,4-dichlorophenoxyacetic acid (2,4-D) and α,α,α-trifluoro-2, 6-dinitro-N,N-dipropyl-p-toluidine (trifluralin); they were then plated on media specific for bacteria, fungi, and actinomycetes. The actinomycete colony count in the trifluralin-treated plot was greater than the control, but the same as the control in the 2,4-D-treated plot. The bacterial count was lower in both treated plots. Fungal colonies in the trifluralin-treated plots were greater than the control, but not different from the control in the 2,4-D-treated plot. PMID:5437308

A High-Speed, Event-Driven, Active Pixel Sensor Readout for Photon-Counting Microchannel Plate Detectors

NASA Technical Reports Server (NTRS)

Kimble, Randy A.; Pain, Bedabrata; Norton, Timothy J.; Haas, J. Patrick; Oegerle, William R. (Technical Monitor)

2002-01-01

Silicon array readouts for microchannel plate intensifiers offer several attractive features. In this class of detector, the electron cloud output of the MCP intensifier is converted to visible light by a phosphor; that light is then fiber-optically coupled to the silicon array. In photon-counting mode, the resulting light splashes on the silicon array are recognized and centroided to fractional pixel accuracy by off-chip electronics. This process can result in very high (MCP-limited) spatial resolution while operating at a modest MCP gain (desirable for dynamic range and long term stability). The principal limitation of intensified CCD systems of this type is their severely limited local dynamic range, as accurate photon counting is achieved only if there are not overlapping event splashes within the frame time of the device. This problem can be ameliorated somewhat by processing events only in pre-selected windows of interest of by using an addressable charge injection device (CID) for the readout array. We are currently pursuing the development of an intriguing alternative readout concept based on using an event-driven CMOS Active Pixel Sensor. APS technology permits the incorporation of discriminator circuitry within each pixel. When coupled with suitable CMOS logic outside the array area, the discriminator circuitry can be used to trigger the readout of small sub-array windows only when and where an event splash has been detected, completely eliminating the local dynamic range problem, while achieving a high global count rate capability and maintaining high spatial resolution. We elaborate on this concept and present our progress toward implementing an event-driven APS readout.

A High-Speed, Event-Driven, Active Pixel Sensor Readout for Photon-Counting Microchannel Plate Detectors

NASA Technical Reports Server (NTRS)

Kimble, Randy A.; Pain, B.; Norton, T. J.; Haas, P.; Fisher, Richard R. (Technical Monitor)

2001-01-01

Silicon array readouts for microchannel plate intensifiers offer several attractive features. In this class of detector, the electron cloud output of the MCP intensifier is converted to visible light by a phosphor; that light is then fiber-optically coupled to the silicon array. In photon-counting mode, the resulting light splashes on the silicon array are recognized and centroided to fractional pixel accuracy by off-chip electronics. This process can result in very high (MCP-limited) spatial resolution for the readout while operating at a modest MCP gain (desirable for dynamic range and long term stability). The principal limitation of intensified CCD systems of this type is their severely limited local dynamic range, as accurate photon counting is achieved only if there are not overlapping event splashes within the frame time of the device. This problem can be ameliorated somewhat by processing events only in pre-selected windows of interest or by using an addressable charge injection device (CID) for the readout array. We are currently pursuing the development of an intriguing alternative readout concept based on using an event-driven CMOS Active Pixel Sensor. APS technology permits the incorporation of discriminator circuitry within each pixel. When coupled with suitable CMOS logic outside the array area, the discriminator circuitry can be used to trigger the readout of small sub-array windows only when and where an event splash has been detected, completely eliminating the local dynamic range problem, while achieving a high global count rate capability and maintaining high spatial resolution. We elaborate on this concept and present our progress toward implementing an event-driven APS readout.

Reduced bacterial colony count of anaerobic bacteria is associated with a worsening in lung clearance index and inflammation in cystic fibrosis.

PubMed

O'Neill, Katherine; Bradley, Judy M; Johnston, Elinor; McGrath, Stephanie; McIlreavey, Leanne; Rowan, Stephen; Reid, Alastair; Bradbury, Ian; Einarsson, Gisli; Elborn, J Stuart; Tunney, Michael M

2015-01-01

Anaerobic bacteria have been identified in abundance in the airways of cystic fibrosis (CF) subjects. The impact their presence and abundance has on lung function and inflammation is unclear. The aim of this study was to investigate the relationship between the colony count of aerobic and anaerobic bacteria, lung clearance index (LCI), spirometry and C-Reactive Protein (CRP) in patients with CF. Sputum and blood were collected from CF patients at a single cross-sectional visit when clinically stable. Community composition and bacterial colony counts were analysed using extended aerobic and anaerobic culture. Patients completed spirometry and a multiple breath washout (MBW) test to obtain LCI. An inverse correlation between colony count of aerobic bacteria (n = 41, r = -0.35; p = 0.02), anaerobic bacteria (n = 41, r = -0.44, p = 0.004) and LCI was observed. There was an inverse correlation between colony count of anaerobic bacteria and CRP (n = 25, r = -0.44, p = 0.03) only. The results of this study demonstrate that a lower colony count of aerobic and anaerobic bacteria correlated with a worse LCI. A lower colony count of anaerobic bacteria also correlated with higher CRP levels. These results indicate that lower abundance of aerobic and anaerobic bacteria may reflect microbiota disruption and disease progression in the CF lung.

Bacterial toxicity/compatibility of platinum nanospheres, nanocuboids and nanoflowers

PubMed Central

Gopal, Judy; Hasan, Nazim; Manikandan, M.; Wu, Hui-Fen

2013-01-01

For the first time, we have investigated the bacterial toxicity or compatibility properties of Pt nanoparticles (NPs) with different sizes (P1, P2, P3, P4 and P5). The bacterio-toxic or compatible properties of these five different sized Pt NPs with the clinical pathogen, Pseudomonas aeruginosa were explored by many analytical methods such as the conventional plate count method, matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS), fluorescence microscopy and fluorescence sensoring techniques. The results revealed that the 1–3 nm sized (P1 and P2) Pt NPs showed bacterio-toxic properties while the 4–21 nm (P3, P4 and P5) Pt NPs exhibited bacterio-compatible properties. This is the first study which reports the bacterial toxicity of Pt NPs. The information released from this study is significantly important to future clinical, medical, biological and biomedical applications of Pt NPs. PMID:23405274

A field study evaluation of Petrifilm™ plates as a 24-h rapid diagnostic test for clinical mastitis on a dairy farm.

PubMed

Mansion-de Vries, Elisabeth Maria; Knorr, Nicole; Paduch, Jan-Hendrik; Zinke, Claudia; Hoedemaker, Martina; Krömker, Volker

2014-03-01

Clinical mastitis is one of the most common and expensive diseases of dairy cattle. To make an informed treatment decision, it is important to know the causative pathogen. However, no detection of bacterial growth can be made in approximately 30% of all clinical cases of mastitis. Before selecting the treatment regimen, it is important to know whether the mastitis-causing pathogen (MCP) is Gram-positive or Gram-negative. The aim of this field study was to investigate whether using two 3M Petrifilm™ products on-farm (which conveys a higher degree of sample freshness but also bears a higher risk for contamination than working in a lab) as 24-h rapid diagnostic of clinical mastitis achieved results that were comparable to the conventional microbiological diagnostic method. AerobicCount (AC)-Petrifilm™ and ColiformCount (CC)-Petrifilm™ were used to identify the total bacterial counts and Gram-negative bacteria in samples from clinical mastitis cases, respectively. Missing growth on both plates was classified as no bacterial detection. Growth only on the AC-Petrifilm™ was assessed as Gram-positive, and growth on both Petrifilm™ plates was assessed as Gram-negative bacterial growth. Additionally, milk samples were analysed by conventional microbiological diagnostic method on aesculin blood agar as a reference method. Overall, 616 samples from clinical mastitis cases were analysed. Using the reference method, Gram-positive and Gram-negative bacteria, mixed bacterial growth, contaminated samples and yeast were determined in 32.6%, 20.0%, 2.5%, 14.1% and 1.1% of the samples, respectively. In 29.7% of the samples, microbiological growth could not be identified. Using the Petrifilm™ concept, bacterial growth was detected in 59% of the culture-negative samples. The sensitivity of the Petrifilm™ for Gram-positive and Gram-negative MCP was 85.2% and 89.9%, respectively. The specificity was 75.4% for Gram-positive and 88.4% for Gram-negative MCP. For the culture

Development of a single-photon-counting camera with use of a triple-stacked micro-channel plate.

PubMed

Yasuda, Naruomi; Suzuki, Hitoshi; Katafuchi, Tetsuro

2016-01-01

At the quantum-mechanical level, all substances (not merely electromagnetic waves such as light and X-rays) exhibit wave–particle duality. Whereas students of radiation science can easily understand the wave nature of electromagnetic waves, the particle (photon) nature may elude them. Therefore, to assist students in understanding the wave–particle duality of electromagnetic waves, we have developed a photon-counting camera that captures single photons in two-dimensional images. As an image intensifier, this camera has a triple-stacked micro-channel plate (MCP) with an amplification factor of 10(6). The ultra-low light of a single photon entering the camera is first converted to an electron through the photoelectric effect on the photocathode. The electron is intensified by the triple-stacked MCP and then converted to a visible light distribution, which is measured by a high-sensitivity complementary metal oxide semiconductor image sensor. Because it detects individual photons, the photon-counting camera is expected to provide students with a complete understanding of the particle nature of electromagnetic waves. Moreover, it measures ultra-weak light that cannot be detected by ordinary low-sensitivity cameras. Therefore, it is suitable for experimental research on scintillator luminescence, biophoton detection, and similar topics.

High level bacterial contamination of secondary school students’ mobile phones

PubMed Central

Kõljalg, Siiri; Mändar, Rando; Sõber, Tiina; Rööp, Tiiu; Mändar, Reet

2017-01-01

Introduction While contamination of mobile phones in the hospital has been found to be common in several studies, little information about bacterial abundance on phones used in the community is available. Our aim was to quantitatively determine the bacterial contamination of secondary school students’ mobile phones. Methods Altogether 27 mobile phones were studied. The contact plate method and microbial identification using MALDI-TOF mass spectrometer were used for culture studies. Quantitative PCR reaction for detection of universal 16S rRNA, Enterococcus faecalis 16S rRNA and Escherichia coli allantoin permease were performed, and the presence of tetracycline (tetA, tetB, tetM), erythromycin (ermB) and sulphonamide (sul1) resistance genes was assessed. Results We found a high median bacterial count on secondary school students’ mobile phones (10.5 CFU/cm2) and a median of 17,032 bacterial 16S rRNA gene copies per phone. Potentially pathogenic microbes (Staphylococcus aureus, Acinetobacter spp., Pseudomonas spp., Bacillus cereus and Neisseria flavescens) were found among dominant microbes more often on phones with higher percentage of E. faecalis in total bacterial 16S rRNA. No differences in contamination level or dominating bacterial species between phone owner’s gender and between phone types (touch screen/keypad) were found. No antibiotic resistance genes were detected on mobile phone surfaces. Conclusion Quantitative study methods revealed high level bacterial contamination of secondary school students’ mobile phones. PMID:28626737

Application of Microbiological Method Direct Epifluorescence Filter Techique/Aerobic Plate Count Agar in the Identification of Irradiated Herbs and Spices

PubMed Central

Di Schiavi, Maria Teresa; Foti, Marina; Mosconi, Maria Cristina; Mattiolo, Giuseppina; Cavallina, Roberta

2014-01-01

Irradiation is a preservation technology used to improve the safety and hygienic quality of food. Aim of this study was to assess the applicability and validity of the microbiological screening method direct epifluorescence filter technique (DEFT)/aerobic plate count (APC) (EN 13783:2001) for the identification of irradiated herbs and spices. Tests on non-irradiated and irradiated samples of dried herbs and spices were performed. The method was based on the comparison of APC and count obtained using DEFT. In accordance with the standard reference, this method is not applicable to samples with APC<103 colony forming units (CFU)/g and this is its main limit. The results obtained in our laboratories showed that in 50% of cases of non-irradiated samples and in 96% of the samples treated with ionising radiation, the method was not applicable due to a value of CFU/g <103. PMID:27800348

Reduced Bacterial Colony Count of Anaerobic Bacteria Is Associated with a Worsening in Lung Clearance Index and Inflammation in Cystic Fibrosis

PubMed Central

Bradley, Judy M.; Johnston, Elinor; McGrath, Stephanie; McIlreavey, Leanne; Rowan, Stephen; Reid, Alastair; Bradbury, Ian; Einarsson, Gisli

2015-01-01

Anaerobic bacteria have been identified in abundance in the airways of cystic fibrosis (CF) subjects. The impact their presence and abundance has on lung function and inflammation is unclear. The aim of this study was to investigate the relationship between the colony count of aerobic and anaerobic bacteria, lung clearance index (LCI), spirometry and C-Reactive Protein (CRP) in patients with CF. Sputum and blood were collected from CF patients at a single cross-sectional visit when clinically stable. Community composition and bacterial colony counts were analysed using extended aerobic and anaerobic culture. Patients completed spirometry and a multiple breath washout (MBW) test to obtain LCI. An inverse correlation between colony count of aerobic bacteria (n = 41, r = -0.35; p = 0.02), anaerobic bacteria (n = 41, r = -0.44, p = 0.004) and LCI was observed. There was an inverse correlation between colony count of anaerobic bacteria and CRP (n = 25, r = -0.44, p = 0.03) only. The results of this study demonstrate that a lower colony count of aerobic and anaerobic bacteria correlated with a worse LCI. A lower colony count of anaerobic bacteria also correlated with higher CRP levels. These results indicate that lower abundance of aerobic and anaerobic bacteria may reflect microbiota disruption and disease progression in the CF lung. PMID:25992575

Method of detecting and counting bacteria

NASA Technical Reports Server (NTRS)

Picciolo, G. L.; Chappelle, E. W. (Inventor)

1976-01-01

An improved method is provided for determining bacterial levels, especially in samples of aqueous physiological fluids. The method depends on the quantitative determination of bacterial adenosine triphosphate (ATP) in the presence of nonbacterial ATP. The bacterial ATP is released by cell rupture and is measured by an enzymatic bioluminescent assay. A concentration technique is included to make the method more sensitive. It is particularly useful where the fluid to be measured contains an unknown or low bacteria count.

Key Issues Concerning Biolog Use for Aerobic and Anaerobic Freshwater Bacterial Community-Level Physiological Profiling

NASA Astrophysics Data System (ADS)

Christian, Bradley W.; Lind, Owen T.

2006-06-01

Bacterial heterotrophy in aquatic ecosystems is important in the overall carbon cycle. Biolog MicroPlates provide information into the metabolic potential of bacteria involved in carbon cycling. Specifically, Biolog EcoPlatesTM were developed with ecologically relevant carbon substrates to allow investigators to measure carbon substrate utilization patterns and develop community-level physiological profiles from natural bacterial assemblages. However, understanding of the functionality of these plates in freshwater research is limited. We explored several issues of EcoPlate use for freshwater bacterial assemblages including inoculum density, incubation temperature, non-bacterial color development, and substrate selectivity. Each of these has various effects on plate interpretation. We offer suggestions and techniques to resolve these interpretation issues. Lastly we propose a technique to allow EcoPlate use in anaerobic freshwater bacterial studies.

Microbial Air Quality and Bacterial Surface Contamination in Ambulances During Patient Services

PubMed Central

Luksamijarulkul, Pipat; Pipitsangjan, Sirikun

2015-01-01

Objectives We sought to assess microbial air quality and bacterial surface contamination on medical instruments and the surrounding areas among 30 ambulance runs during service. Methods We performed a cross-sectional study of 106 air samples collected from 30 ambulances before patient services and 212 air samples collected during patient services to assess the bacterial and fungal counts at the two time points. Additionally, 226 surface swab samples were collected from medical instrument surfaces and the surrounding areas before and after ambulance runs. Groups or genus of isolated bacteria and fungi were preliminarily identified by Gram’s stain and lactophenol cotton blue. Data were analyzed using descriptive statistics, t-test, and Pearson’s correlation coefficient with a p-value of less than 0.050 considered significant. Results The mean and standard deviation of bacterial and fungal counts at the start of ambulance runs were 318±485cfu/m3 and 522±581cfu/m3, respectively. Bacterial counts during patient services were 468±607cfu/m3 and fungal counts were 656±612cfu/m3. Mean bacterial and fungal counts during patient services were significantly higher than those at the start of ambulance runs, p=0.005 and p=0.030, respectively. For surface contamination, the overall bacterial counts before and after patient services were 0.8±0.7cfu/cm2 and 1.3±1.1cfu/cm2, respectively (p<0.001). The predominant isolated bacteria and fungi were Staphylococcus spp. and Aspergillus spp., respectively. Additionally, there was a significantly positive correlation between bacterial (r=0.3, p<0.010) and fungal counts (r=0.2, p=0.020) in air samples and bacterial counts on medical instruments and allocated areas. Conclusions This study revealed high microbial contamination (bacterial and fungal) in ambulance air during services and higher bacterial contamination on medical instrument surfaces and allocated areas after ambulance services compared to the start of ambulance runs

Microbial air quality and bacterial surface contamination in ambulances during patient services.

PubMed

Luksamijarulkul, Pipat; Pipitsangjan, Sirikun

2015-03-01

We sought to assess microbial air quality and bacterial surface contamination on medical instruments and the surrounding areas among 30 ambulance runs during service. We performed a cross-sectional study of 106 air samples collected from 30 ambulances before patient services and 212 air samples collected during patient services to assess the bacterial and fungal counts at the two time points. Additionally, 226 surface swab samples were collected from medical instrument surfaces and the surrounding areas before and after ambulance runs. Groups or genus of isolated bacteria and fungi were preliminarily identified by Gram's stain and lactophenol cotton blue. Data were analyzed using descriptive statistics, t-test, and Pearson's correlation coefficient with a p-value of less than 0.050 considered significant. The mean and standard deviation of bacterial and fungal counts at the start of ambulance runs were 318±485cfu/m(3) and 522±581cfu/m(3), respectively. Bacterial counts during patient services were 468±607cfu/m(3) and fungal counts were 656±612cfu/m(3). Mean bacterial and fungal counts during patient services were significantly higher than those at the start of ambulance runs, p=0.005 and p=0.030, respectively. For surface contamination, the overall bacterial counts before and after patient services were 0.8±0.7cfu/cm(2) and 1.3±1.1cfu/cm(2), respectively (p<0.001). The predominant isolated bacteria and fungi were Staphylococcus spp. and Aspergillus spp., respectively. Additionally, there was a significantly positive correlation between bacterial (r=0.3, p<0.010) and fungal counts (r=0.2, p=0.020) in air samples and bacterial counts on medical instruments and allocated areas. This study revealed high microbial contamination (bacterial and fungal) in ambulance air during services and higher bacterial contamination on medical instrument surfaces and allocated areas after ambulance services compared to the start of ambulance runs. Additionally, bacterial and

The fixation effect of a silk fibroin-bacterial cellulose composite plate in segmental defects of the zygomatic arch: an experimental study.

PubMed

Lee, Jung Min; Kim, Ji Heui; Lee, Ok Joo; Park, Chan Hum

2013-06-01

Bioresorbable fixation systems have been popular for the treatment of facial fractures. However, their mechanical properties are uncertain and complications have been reported. To overcome these problems, we developed a bioresorbable fixation plate using a composite of silk fibroin and bacterial cellulose (SF-BC) with biodegradability and increased biocompatibility. To investigate the regenerative effect of the bioresorbable SF-BC fixation plate on zygomatic arch defects in rats. In vivo animal study. The SF-BC composite plate had a tensile strength similar to that of a polylactic acid plate and a tight, pore-free microstructure. Bilateral segmental bone defects (2 mm in length) were created in the zygomatic arches of adult rats. One side was fixed with the SF-BC composite plate, and the other side was left without fixation. Academic research laboratory. Fifteen adult Sprague-Dawley rats. Fixation of the zygomatic arch defect with the SF-BC composite plate. Micro-computed tomography and histological evaluation of bone samples. Gross inspection revealed no specific complication. At 1, 2, 4, and 8 postoperative weeks, the zygomatic arches were explored by micro-computed tomography and histological examination. Control sides did not heal completely and showed bony degeneration and necrosis during the 8-week follow-up. However, we observed new bone formation in sides treated with the SF-BC composite plate, and bony defects were completely healed within 8 weeks. The SF-BC composite plate is a potential candidate for a new bioresorbable fixation system. Our composite material could considerably shorten bone regeneration time. Additional study of the control of biodegradability and mechanical properties of SF-BC composite plates and a comparative study with the resorbable plates currently in use should be undertaken.

Pseudoalteromonas spp. Serve as Initial Bacterial Attractants in Mesocosms of Coastal Waters but Have Subsequent Antifouling Capacity in Mesocosms and when Embedded in Paint

PubMed Central

Bernbom, Nete; Ng, Yoke Yin; Olsen, Stefan Møller

2013-01-01

The purpose of the present study was to determine if the monoculture antifouling effect of several pigmented pseudoalteromonads was retained in in vitro mesocosm systems using natural coastal seawater and when the bacteria were embedded in paint used on surfaces submerged in coastal waters. Pseudoalteromonas piscicida survived on a steel surface and retained antifouling activity for at least 53 days in sterile seawater, whereas P. tunicata survived and had antifouling activity for only 1 week. However, during the first week, all Pseudoalteromonas strains facilitated rather than prevented bacterial attachment when used to coat stainless steel surfaces and submerged in mesocosms with natural seawater. The bacterial density on surfaces coated with sterile growth medium was 105 cells/cm2 after 7 days, whereas counts on surfaces precoated with Pseudoalteromonas were significantly higher, at 106 to 108 cells/cm2. However, after 53 days, seven of eight Pseudoalteromonas strains had reduced total bacterial adhesion compared to the control. P. piscicida, P. antarctica, and P. ulvae remained on the surface, at levels similar to those in the initial coating, whereas P. tunicata could not be detected. Larger fouling organisms were observed on all plates precoated with Pseudoalteromonas; however, plates coated only with sterile growth medium were dominated by a bacterial biofilm. Suspensions of a P. piscicida strain and a P. tunicata strain were incorporated into ship paints (Hempasil x3 87500 and Hempasil 77500) used on plates that were placed at the Hempel A/S test site in Jyllinge Harbor. For the first 4 months, no differences were observed between control plates and treated plates, but after 5 to 6 months, the control plates were more fouled than the plates with pseudoalteromonad-based paint. Our study demonstrates that no single laboratory assay can predict antifouling effects and that a combination of laboratory and real-life methods must be used to determine the

Prevention of Bacterial Biofilms Formation on Urinary Catheter by Selected Plant Extracts.

PubMed

Adesina, T D; Nwinyi, O C; Olugbuyiro, J A O

2015-02-01

In this study, we investigated the feasibility of using Psidium guajava, Mangifera indica and Ocimum gratissimum leaf extracts in preventing Escherichia coli biofilm formation. The plants extractions were done with methanol under cold extraction. The various concentrations 5.0, 10.0 and 20.0 mg mL(-1) were used to coat 63 catheters under mild heat from water bath. Biofilm formation on the catheter was induced using cultures of E. coli. Biofilm formation was evaluated using aerobic plate count and turbidity at 600 nm. From the obtained results, Psidium guajava, Mangifera indica and Ocimum gratissimum delayed the onset of biofilm formation for a week. Ocimum gratissimum coated catheter had the highest inhibitory effect at 5.0, 10.0 and 20.0 mg mL(-1) with bacterial count ranging from 2.2 x 10(5)-7.0 x 10(4) and 5.7 x 10(5)-3.7 x10(5) for 120 and 128 h, respectively. The Psidium guajava coated catheter had the lowest inhibitory effect at 5.0, 10.0 and 20.0 mg mL(-1), with bacterial count ranging between 4.3 x 10(5)-1.9 x 10(3) and 7.7 x 10(5)-3.8 x 10(5) for 120 and 128 h, respectively. Despite the antimicrobial activities, the differences in the activity of these plant extracts were statistically not significant (p < 0.05).

Risk factors associated with bulk tank standard plate count, bulk tank coliform count, and the presence of Staphylococcus aureus on organic and conventional dairy farms in the United States.

PubMed

Cicconi-Hogan, K M; Gamroth, M; Richert, R; Ruegg, P L; Stiglbauer, K E; Schukken, Y H

2013-01-01

The purpose of this study was to assess the association of bulk tank milk standard plate counts, bulk tank coliform counts (CC), and the presence of Staphylococcus aureus in bulk tank milk with various management and farm characteristics on organic and conventional dairy farms throughout New York, Wisconsin, and Oregon. Data from size-matched organic farms (n=192), conventional nongrazing farms (n=64), and conventional grazing farms (n=36) were collected at a single visit for each farm. Of the 292 farms visited, 290 bulk tank milk samples were collected. Statistical models were created using data from all herds in the study, as well as exclusively for the organic subset of herds. Because of incomplete data, 267 of 290 herds were analyzed for total herd modeling, and 173 of 190 organic herds were analyzed for the organic herd modeling. Overall, more bulk tanks from organic farms had Staph. aureus cultured from them (62% of organic herds, 42% conventional nongrazing herds, and 43% of conventional grazing herds), whereas fewer organic herds had a high CC, defined as ≥50 cfu/mL, than conventional farms in the study. A high standard plate count (×1,000 cfu/mL) was associated with decreased body condition score of adult cows and decreased milk production in both models. Several variables were significant only in the model created using all herds or only in organic herds. The presence of Staph. aureus in the bulk tank milk was associated with fewer people treating mastitis, increased age of housing, and a higher percentage of cows with 3 or fewer teats in both the organic and total herd models. The Staph. aureus total herd model also showed a relationship with fewer first-lactation animals, higher hock scores, and less use of automatic takeoffs at milking. High bulk tank CC was related to feeding a total mixed ration and using natural service in nonlactating heifers in both models. Overall, attentive management and use of outside resources were useful with regard to CC

Reduction effect of bacterial counts by preoperative saline lavage of the stomach in performing laparoscopic and endoscopic cooperative surgery.

PubMed

Mori, Hirohito; Kobara, Hideki; Tsushimi, Takaaki; Fujihara, Shintaro; Nishiyama, Noriko; Matsunaga, Tae; Ayaki, Maki; Yachida, Tatsuo; Tani, Joji; Miyoshi, Hisaaki; Morishita, Asahiro; Masaki, Tsutomu

2014-11-14

To investigate the effects of gastric lavage with 2000 mL of saline in laparoscopic and endoscopic cooperative surgery. Twenty two patients who were diagnosed with a gastric gastrointestinal stromal tumor were enrolled. In former term, irrigations of the stomach were conducted whenever it was necessary, not systematically (Non systemic lavage group). In latter term, the stomach was thoroughly cleaned with 2000 mL of saline using an endoscope with a water jet, and Duodenal balloon occlusion was conducted to prevent refluxed bile and pancreatic juice (Systemic lavage+balloon occlusion group). The gastric wall was sprayed with 20 mL of distilled water, and 20 mL of gastric juice was collected in a sterile tube and submitted for culture. 20 mL of ascites was also collected from the laparoscopic ports and submitted for culture. We compared WBC, CRP, BT between two groups, and verify the reduction effect of bacterial counts in Systemic lavage+balloon occlusion group. WBC count before, 1 d after, and 3 d after laparoscopic and endoscopic cooperative surgery (LECS) were 5060 (95%CI: 4250-9640), 12140 (6050-14110), and 6910 (5320-12520) in Non systemic lavage group, 4400 (3660-7620), 8910 (6480-10980), and 5950 (4840-7860) in Systemic lavage+balloon occlusion group. Significant differences between two groups at the day after LECS (P = 0.029) and the 3 d after LECS (P = 0.042). CRP levels in Non systemic lavage group and in Systemic lavage+balloon occlusion group were significantly different at the day after LECS (P = 0.005) and the 3 d after LECS (P = 0.028). BTs (°C) in Non systemic lavage group and in Systemic lavage+balloon occlusion group were also significantly different at the day after LECS (P = 0.004) and the 3 d after LECS (P = 0.006). In a logarithmic comparison, bacterial load before gastric lavage, after lavage, and ascites culture were 6.08 (95%CI: 4.04-6.97), 0.48 (0-0.85), and 0.21 (0-0.56). The bacterial counts before and after gastric lavage were

Microbiological survey of a South African poultry processing plant.

PubMed

Geornaras, I; de Jesus, A; van Zyl, E; von Holy, A

1995-01-01

Bacterial populations associated with poultry processing were determined on neck skin samples, equipment surfaces and environmental samples by replicate surveys. Aerobic plate counts, Enterobacteriaceae counts, Enterobacteriaceae counts and Pseudomonas counts were performed by standard procedures and the prevalence of Listeria, presumptive Salmonella and Staphylococcus aureus determined. Statistically significant (P < 0.05) increases in counts of all types of bacteria were obtained on product samples as a result of processing. Although bacterial counts on neck skin samples decreased by 0.3 to 0.4 log CFU g-1 after spray washing of carcasses, subsequent spinchilling and packaging of whole carcasses resulted in 0.7 to 1.2 log CFU g-1 increases. Bacterial numbers on equipment surfaces, however, decreased significantly from the "dirty" to the "clean" areas of the abattoir. Transport cages, "rubber fingers", defeathering curtains, shackles and conveyor belts repeatedly showed aerobic plate counts in excess of 5.0 log CFU 25 cm-2. Aerobic plate counts of scald tank and spinchiller water were 2 log CFU ml-1 higher than those of potable water samples. Bacterial numbers of the air in the "dirty" area were higher than those of the "clean" area. Listeria, presumptive Salmonella and Staphylococcus aureus were isolated from 27.6, 51.7 and 24.1% of all product samples, respectively, and Listeria and Staphylococcus aureus were also isolated from selected equipment surfaces.

Phage-mediated counting by the naked eye of miRNA molecules at attomolar concentrations in a Petri dish.

PubMed

Zhou, Xin; Cao, Peng; Zhu, Ye; Lu, Wuguang; Gu, Ning; Mao, Chuanbin

2015-10-01

The ability to count biomolecules such as cancer-biomarker miRNAs with the naked eye is seemingly impossible in molecular diagnostics. Here, we show an ultrasensitive naked-eye-counting strategy for quantifying miRNAs by employing T7 phage-a bacteria-specific virus nanoparticle-as a surrogate. The phage is genetically engineered to become fluorescent and capable of binding a miRNA-capturing gold nanoparticle (GNP) in a one-to-one manner. Target miRNAs crosslink the resultant phage-GNP couple and miRNA-capturing magnetic microparticles, forming a sandwich complex containing equimolar phage and miRNA. The phage is then released from the complex and developed into one macroscopic fluorescent plaque in a Petri dish by plating it in a host bacterial medium. Counting the plaques by the naked eye enables the quantification of miRNAs with detection limits of ∼3 and ∼5 aM for single-target and two-target miRNAs, respectively. This approach offers ultrasensitive and convenient quantification of disease biomarkers by the naked eye.

Phage-mediated counting by the naked eye of miRNA molecules at attomolar concentrations in a Petri dish

NASA Astrophysics Data System (ADS)

Zhou, Xin; Cao, Peng; Zhu, Ye; Lu, Wuguang; Gu, Ning; Mao, Chuanbin

2015-10-01

The ability to count biomolecules such as cancer-biomarker miRNAs with the naked eye is seemingly impossible in molecular diagnostics. Here, we show an ultrasensitive naked-eye-counting strategy for quantifying miRNAs by employing T7 phage--a bacteria-specific virus nanoparticle--as a surrogate. The phage is genetically engineered to become fluorescent and capable of binding a miRNA-capturing gold nanoparticle (GNP) in a one-to-one manner. Target miRNAs crosslink the resultant phage-GNP couple and miRNA-capturing magnetic microparticles, forming a sandwich complex containing equimolar phage and miRNA. The phage is then released from the complex and developed into one macroscopic fluorescent plaque in a Petri dish by plating it in a host bacterial medium. Counting the plaques by the naked eye enables the quantification of miRNAs with detection limits of ~3 and ~5 aM for single-target and two-target miRNAs, respectively. This approach offers ultrasensitive and convenient quantification of disease biomarkers by the naked eye.

Association between bacterial survival and free chlorine concentration during commercial fresh-cut produce wash operation.

PubMed

Luo, Yaguang; Zhou, Bin; Van Haute, Sam; Nou, Xiangwu; Zhang, Boce; Teng, Zi; Turner, Ellen R; Wang, Qin; Millner, Patricia D

2018-04-01

Determining the minimal effective free chlorine (FC) concentration for preventing pathogen survival and cross-contamination during produce washing is critical for developing science- and risk-based food safety practices. The correlation between dynamic FC concentrations and bacterial survival was investigated during commercial washing of chopped Romaine lettuce, shredded Iceberg lettuce, and diced cabbage as pathogen inoculation study during commercial operation is not feasible. Wash water was sampled every 30 min and assayed for organic loading, FC, and total aerobic mesophilic bacteria after chlorine neutralization. Water turbidity, chemical oxygen demand, and total dissolved solids increased significantly over time, with more rapid increases in diced cabbage water. Combined chlorine increased consistently while FC fluctuated in response to rates of chlorine dosing, product loading, and water replenishment. Total bacterial survival showed a strong correlation with real-time FC concentration. Under approximately 10 mg/L, increasing FC significantly reduced the frequency and population of surviving bacteria detected. Increasing FC further resulted in the reduction of the aerobic plate count to below the detection limit (50 CFU/100 mL), except for a few sporadic positive samples with low cell counts. This study confirms that maintaining at least 10 mg/L FC in wash water strongly reduced the likelihood of bacterial survival and thus potential cross contamination of washed produce. Published by Elsevier Ltd.

Nondestructive detection of total viable count changes of chilled pork in high oxygen storage condition based on hyperspectral technology

NASA Astrophysics Data System (ADS)

Zheng, Xiaochun; Peng, Yankun; Li, Yongyu; Chao, Kuanglin; Qin, Jianwei

2017-05-01

The plate count method is commonly used to detect the total viable count (TVC) of bacteria in pork, which is timeconsuming and destructive. It has also been used to study the changes of the TVC in pork under different storage conditions. In recent years, many scholars have explored the non-destructive methods on detecting TVC by using visible near infrared (VIS/NIR) technology and hyperspectral technology. The TVC in chilled pork was monitored under high oxygen condition in this study by using hyperspectral technology in order to evaluate the changes of total bacterial count during storage, and then evaluate advantages and disadvantages of the storage condition. The VIS/NIR hyperspectral images of samples stored in high oxygen condition was acquired by a hyperspectral system in range of 400 1100nm. The actual reference value of total bacteria was measured by standard plate count method, and the results were obtained in 48 hours. The reflection spectra of the samples are extracted and used for the establishment of prediction model for TVC. The spectral preprocessing methods of standard normal variate transformation (SNV), multiple scatter correction (MSC) and derivation was conducted to the original reflectance spectra of samples. Partial least squares regression (PLSR) of TVC was performed and optimized to be the prediction model. The results show that the near infrared hyperspectral technology based on 400-1100nm combined with PLSR model can describe the growth pattern of the total bacteria count of the chilled pork under the condition of high oxygen very vividly and rapidly. The results obtained in this study demonstrate that the nondestructive method of TVC based on NIR hyperspectral has great potential in monitoring of edible safety in processing and storage of meat.

Bioorganic Fertilizer Enhances Soil Suppressive Capacity against Bacterial Wilt of Tomato

PubMed Central

Liu, Shuangri; Chai, Rushan; Huang, Weiqing; Liu, Xingxing; Tang, Caixian; Zhang, Yongsong

2015-01-01

Tomato bacterial wilt caused by Ralstonia solanacearum is one of the most destructive soil-borne diseases. Many strategies have been taken to improve soil suppressiveness against this destructive disease, but limited success has been achieved. In this study, a novel bioorganic fertilizer revealed a higher suppressive ability against bacterial wilt compared with several soil management methods in the field over four growing seasons from March 2011 to July 2013. The application of the bioorganic fertilizer significantly (P<0.05) reduced disease incidence of tomato and increased fruit yields in four independent trials. The association among the level of disease incidence, soil physicochemical and biological properties was investigated. The soil treated with the bioorganic fertilizer increased soil pH value, electric conductivity, organic carbon, NH4 +-N, NO3 --N and available K content, microbial activities and microbial biomass carbon content, which were positively related with soil suppressiveness. Bacterial and actinomycete populations assessed using classical plate counts were highest, whereas R. solanacearum and fungal populations were lowest in soil applied with the bioorganic fertilizer. Microbial community diversity and richness were assessed using denaturing gel gradient electrophoresis profile analysis. The soil treated with the bioorganic fertilizer exhibited higher bacterial community diversity but lower fungal community diversity. Redundancy analysis showed that bacterial community diversity and richness negatively related with bacterial wilt suppressiveness, while fungal community richness positively correlated with R. solanacearum population. We concluded that the alteration of soil physicochemical and biological properties in soil treated with the bioorganic fertilizer induced the soil suppressiveness against tomato bacterial wilt. PMID:25830639

Profile of selected bacterial counts and Salmonella prevalence on raw poultry in a poultry slaughter establishment.

PubMed

James, W O; Williams, W O; Prucha, J C; Johnston, R; Christensen, W

1992-01-01

The USDA Food Safety and Inspection Service determined populations of bacteria on poultry during processing at a slaughter plant in Puerto Rico in November and December 1987. The plant was selected because of its management's willingness to support important changes in equipment and processing procedures. The plant was representative of modern slaughter facilities. Eight-hundred samples were collected over 20 consecutive 8-hour days of operation from 5 sites in the processing plant. Results indicated that slaughter, dressing, and chilling practices significantly decreased the bacterial contamination on poultry carcasses, as determined by counts of aerobic bacteria, Enterobacteriaceae, and Escherichia coli. Salmonella was not enumerated; rather, it was determined to be present or absent by culturing almost the entire rinse. The prevalence of Salmonella in the study decreased during evisceration, then increased during immersion chilling.

Blue light (470 nm) effectively inhibits bacterial and fungal growth.

PubMed

De Lucca, A J; Carter-Wientjes, C; Williams, K A; Bhatnagar, D

2012-12-01

Blue light (470 nm) LED antimicrobial properties were studied alone against bacteria and with or without the food grade photosensitizer, erythrosine (ERY) against filamentous fungi. Leuconostoc mesenteroides (LM), Bacillus atrophaeus (BA) or Pseudomonas aeruginosa (PA) aliquots were exposed on nutrient agar plates to Array 1 (AR1, 0·2 mW cm(-2)) or Array 2 (AR2, 80 mW cm(-2)), which emitted impure or pure blue light (0-300 J cm(-2)), respectively. Inoculated control (room light only) plates were incubated (48 h) and colonies enumerated. The antifungal properties of blue light combined with ERY (11·4 and 22·8 μmol l(-1)) on Penicillium digitatum (PD) and Fusarium graminearum (FG) conidia were determined. Conidial controls consisted of: no light, room light-treated conidia and ERY plus room light. Light-treated (ERY + blue light) conidial samples were exposed only to AR2 (0-100 J cm(-2)), aliquots spread on potato dextrose agar plates, incubated (48 h, 30°C) and colonies counted. Blue light alone significantly reduced bacterial and FG viability. Combined with ERY, it significantly reduced PD viability. Blue light is lethal to bacteria and filamentous fungi although effectiveness is dependent on light purity, energy levels and microbial genus. Light from two arrays of different blue LEDs significantly reduced bacterial (Leuconostoc mesenteroides, Bacillus atrophaeus and Pseudomonas aeruginosa) viabilities. Significant in vitro viability loss was observed for the filamentous fungi, Penicillium digitatum and Fusarium graminearum when exposed to pure blue light only plus a photosensitizer. F. graminearum viability was significantly reduced by blue light alone. Results suggest that (i) the amount of significant loss in bacterial viability observed for blue light that is pure or with traces of other wavelengths is genus dependent and (ii) depending on fungal genera, pure blue light is fungicidal with or without a photosensitizer. © 2012 The Society for

Superiority of a functional leukocyte adhesiveness/aggregation test over the white blood cell count to discriminate between mild and significant inflammatory response in patients with acute bacterial infections.

PubMed

Rogowski, Ori; Rotstein, Rivka; Zeltzer, David; Misgav, Sarit; Justo, Daniel; Avitzour, Daniel; Mardi, Tamar; Serov, Jacob; Arber, Nadir; Berliner, Shlomo; Shapira, Itzhak

2002-01-01

Electronic cell counters may underestimate the white blood cell count (WBCC) in the presence of aggregated leukocytes. In the present study we focused on the possibility of using a functional, as opposed to an anatomic, count to circumvent this eventual underestimation. A model of bacterial infection was used because of the importance of leukocytosis in the physician's clinical decision-making process. There were 35 patients with low C-reactive protein (CRP) concentrations (0.5-4.9 mg/dL), 45 with intermediate (5-9.9 mg/dL), and 120 with relatively high (>10 mg/dL) CRP concentrations. A significant (P=0.008) difference was noted between the state of leukocyte adhesiveness/aggregation in the peripheral blood of individuals with low CRP concentrations (3.5%+/-4.3%) and those with high CRP concentrations (7.4%+/-8%), while there was no significant difference in the respective number of WBCs per cubic millimeter (cmm) (11,600 +/- 5,500 and 14,000 +/- 7,200, respectively). We raise the possibility that a functional test might be superior over an anatomic count in patients with acute bacterial infection and a significant acute phase response. Copyright 2002 Wiley-Liss, Inc.

Phage-based biomolecular filter for the capture of bacterial pathogens in liquid streams

NASA Astrophysics Data System (ADS)

Du, Songtao; Chen, I.-Hsuan; Horikawa, Shin; Lu, Xu; Liu, Yuzhe; Wikle, Howard C.; Suh, Sang Jin; Chin, Bryan A.

2017-05-01

This paper investigates a phage-based biomolecular filter that enables the evaluation of large volumes of liquids for the presence of small quantities of bacterial pathogens. The filter is a planar arrangement of phage-coated, strip-shaped magnetoelastic (ME) biosensors (4 mm × 0.8 mm × 0.03 mm), magnetically coupled to a filter frame structure, through which a liquid of interest flows. This "phage filter" is designed to capture specific bacterial pathogens and allow non-specific debris to pass, eliminating the common clogging issue in conventional bead filters. ANSYS Maxwell was used to simulate the magnetic field pattern required to hold ME biosensors densely and to optimize the frame design. Based on the simulation results, a phage filter structure was constructed, and a proof-in-concept experiment was conducted where a Salmonella solution of known concentration were passed through the filter, and the number of captured Salmonella was quantified by plate counting.

Microbial contamination in intraoral phosphor storage plates: the dilemma.

PubMed

de Souza, Tricia Murielly Pereira Andrade; de Castro, Ricardo Dias; de Vasconcelos, Laís César; Pontual, Andréa Dos Anjos; de Moraes Ramos Perez, Flávia Maria; Pontual, Maria Luiza Dos Anjos

2017-01-01

The aims of this study were to evaluate microbial contamination in phosphor storage plates in dental radiology services and discuss the possible origin of this contamination. The sample comprised 50 phosphor plates: 14 plates from service A, 30 from service B, and 6 in the control group, consisting of plates never used. Damp sterile swabs were rubbed on the phosphor plates, and then transferred to tests tubes containing sterile saline solution. Serial dilutions were made, and then inoculated in triplicate on Mueller Hinton agar plates and incubated at 37 °C/48 h, before counting the colony-forming units (CFU). The samples were also seeded in brain-heart infusion medium to confirm contamination by turbidity of the culture medium. All solutions, turbid and clean, were seeded in selective and non-selective media. At service A and B, 50 and 73.3 % of the phosphor plates were contaminated, respectively. This contamination was mainly due to bacteria of the genus Staphylococcus. CFU counts ranged from 26.4 to 80.0 CFU/plate. Most of the phosphor plates evaluated shown to be contaminated, mainly by Staphylococcus ssp. Quantitatively, this contamination occurred at low levels, possibly arising from handling of the plates. The use of a second plastic barrier may have diminished contamination by microorganisms from the oral cavity. There is a risk of cross-contamination by phosphor storage plates used in dental radiology services.

Laboratory evaluation of 3M Petrifilms and University of Minnesota Bi-plates as potential on-farm tests for clinical mastitis.

PubMed

McCarron, J L; Keefe, G P; McKenna, S L B; Dohoo, I R; Poole, D E

2009-05-01

The objective was to determine test characteristics and compare 2 potential on-farm culture systems for clinical mastitis, the Minnesota Easy Culture System II Bi-plate and Petrifilm. The tests were evaluated using clinically positive mastitic milk samples (n = 282) to determine their ability to differentiate appropriate treatment groups; all cases that had gram-positive growth were considered treatment candidates (n = 161), whereas cases that grew gram-negative organisms only or yielded no bacterial growth were classified as no treatment (n = 121). For Petrifilm, both undiluted and 1:10 diluted milk samples were used. To create treatment categories, 2 types of Petrifilms were used, Aerobic Count (AC) and Coliform Count (CC). Both Bi-plates and Petrifilms were read after 24 h of incubation. Analysis was conducted at various colony count thresholds for the Petrifilm test system. The combination of Petrifilms that had the highest sensitivity classified a case as gram-negative if there were > or =20 colonies present on the CC. If there were <20 colonies present on the CC and >5 colonies present on the AC, a case would be classified as gram-positive. The Bi-plate had a sensitivity of 97.9% and a specificity of 68.6%. The Petrifilm test system had a sensitivity of 93.8% and a specificity of 70.1%. There was no significant difference in the sensitivities between the tests. All Bi-plates and Petrifilms were read by a laboratory technician and a group of masked readers with limited microbiology training. Kappa values for the masked readers were 0.75 for Bi-plates and 0.84 and 0.86 for AC and CC Petrifilms, respectively. The Bi-plate and Petrifilm were able to successfully categorize clinical cases of mastitis into 2 treatments based on their ability to detect the presence of a gram-positive organism. Neither method had the ability to determine if a sample was contaminated. The results of this study indicate that both tests were able to appropriately categorize cases, which

Factors influencing variation of bulk milk antibiotic residue occurrence, somatic cell count, and total bacterial count in dairy sheep flocks.

PubMed

Gonzalo, C; Carriedo, J A; García-Jimeno, M C; Pérez-Bilbao, M; de la Fuente, L F

2010-04-01

To study the variations of bulk tank milk variables in dairy ewe flocks and to identify the main target practices and flock groups to improve milk quality and safety, a total of 71,228 records of antibiotic residue (AR) and milk yield and 68,781 records of somatic cell count (SCC) and total bacterial count (TBC) were obtained over 5 yr from the same 209 dairy ewe flocks of the Assaf breed belonging to the Consortium for Ovine Promotion of Castilla-León (Spain). Based on a logistic regression model, year, month, semester, SCC, TBC, dry therapy, and milk yield significantly contributed to AR variation. High SCC was associated with increased AR violations. When antibiotic dry therapy was implemented, AR occurrence was higher than when this practice was not used. A polynomial monthly distribution throughout the year was observed for AR occurrence; the highest values were in autumn, coinciding with low milk yields per flock. Yearly occurrences drastically diminished from 2004 (1.36%) to 2008 (0.30%), probably as a result of effective educational programs. The mixed-model ANOVA of factors influencing variation in SCC and TBC indicated that year, month, AR, dry therapy group, milking type, and year interactions were significant variation factors for SCC and TBC; mathematical model accounted for 74.1 and 35.4% of total variance for each variable, respectively. Differences in management and hygiene practice caused significant SCC and TBC variations among flocks and within flocks throughout the 5-yr study. Over time, continuously dry treated flocks showed lower logSCC (5.80) and logTBC (4.92) than untreated (6.10 and 5.18, respectively) or discontinuously dry treated (6.01 and 5.05, respectively) flocks. Continuously dry treated flocks had lower AR occurrences than did discontinuously dry treated flocks. As a whole, AR occurrence and SCC and TBC bulk tank milk variables can be used for monitoring mammary health and milk hygiene and safety in dairy sheep throughout time

Surface Enhanced Raman Spectroscopy for the Rapid Detection and Identification of Microbial Pathogens in Human Serum

DTIC Science & Technology

2014-12-11

and 1 mm depth. Bacterial culture and cell count determination Bacterial species of Acinetobacter baumannii (A. baumannii, ST-3), Escherichia coli...remove all broth components followed by a final resuspension of the pellet in ddH2O back to 1 OD. Cell count was determined by plating the 10 4 , 10 3...10 2 and 10 1 cell dilutions on TSB Nutrient Agar media. Colony forming units (CFU) were counted the following day to confirm bacterial species

The utility of biomarkers in differentiating bacterial from non-bacterial lower respiratory tract infection in hospitalized children: difference of the diagnostic performance between acute pneumonia and bronchitis.

PubMed

Hoshina, Takayuki; Nanishi, Etsuro; Kanno, Shunsuke; Nishio, Hisanori; Kusuhara, Koichi; Hara, Toshiro

2014-10-01

The aim of this study is to investigate the utility of several biomarkers in differentiating bacterial community-acquired lower respiratory tract infection (CA-LRTI) from non-bacterial CA-LRTI in children and the difference of their diagnostic performance between pneumonia and bronchitis. A retrospective cohort study composed of 108 pediatric patients hospitalized for CA-LRTI was performed during 2010-2013. Based on the findings of chest X-ray and sputum samples, patients were divided into 4 categories, group of bacterial pneumonia or bronchitis, and non-bacterial (viral or etiology-unknown) pneumonia or bronchitis. Peripheral white blood cell and neutrophil counts, and serum C-reactive protein (CRP) and procalcitonin (PCT) levels were compared among the 4 groups. Finally, 54 patients were the subject of this study. In the patients with pneumonia, serum CRP and PCT levels were significantly elevated in the group of bacterial pneumonia (CRP: p = 0.02, PCT: p = 0.0008). The area under the receiver operating characteristic curve for PCT for distinguishing between bacterial and non-bacterial pneumonia was the largest, and sensitivity, specificity, positive predictive value and negative predictive value of PCT were best among 4 markers. On the other hand, in the patients with bronchitis, neutrophil count was significantly decreased in non-bacterial bronchitis whereas no significant differences of WBC count, CRP level or PCT level were seen. In conclusion, PCT was the most useful marker to differentiate bacterial pneumonia whereas neutrophil count contributed most to the discrimination of bacterial bronchitis. The diagnostic performance of biomarkers may be different between pneumonia and bronchitis. Copyright © 2014. Published by Elsevier Ltd.

Trace Amounts of Furan-2-Carboxylic Acids Determine the Quality of Solid Agar Plates for Bacterial Culture

PubMed Central

Hara, Shintaro; Isoda, Reika; Tahvanainen, Teemu; Hashidoko, Yasuyuki

2012-01-01

Background Many investigators have recognised that a significant proportion of environmental bacteria exist in a viable but non-culturable state on agar plates, and some researchers have also noticed that some of such bacteria clearly recover their growth on matrices other than agar. However, the reason why agar is unsuitable for the growth of some bacteria has not been addressed. Methodology/Principal Findings According to the guide of a bioassay for swarming inhibition, we identified 5-hydroxymethylfuran-2-carboxylic acid (5-HMFA) and furan-2-carboxylic acid (FA) as factors that inhibit bacterial swarming and likely inhibit extracellular polysaccharide production on agar. The furan-2-carboxylic acids 5-HMFA and FA effectively inhibited the swarming and swimming of several environmental bacteria at concentrations of 1.8 and 2.3 µg L−1 (13 and 21 nmol L−1), respectively, which are equivalent to the concentrations of these compounds in 0.3% agar. On Luria-Bertani (LB) plates containing 1.0% agar that had been previously washed with MeOH, a mixture of 5-HMFA and FA in amounts equivalent to their original concentrations in the unwashed agar repressed the swarming of Escherichia coli K12 strain W3110, a representative swarming bacterium. Conclusions/Significance Agar that contains trace amounts of 5-HMFA and FA inhibits the proliferation of some slow-growing or difficult-to-culture bacteria on the plates, but it is useful for single colony isolation due to the ease of identification of swarmable bacteria as the non-swarmed colonies. PMID:22848437

Evaluation of free-stall mattress bedding treatments to reduce mastitis bacterial growth.

PubMed

Kristula, M A; Dou, Z; Toth, J D; Smith, B I; Harvey, N; Sabo, M

2008-05-01

Bacterial counts were compared in free-stall mattresses and teat ends exposed to 5 treatments in a factorial study design on 1 dairy farm. Mattresses in five 30-cow groups were subjected to 1 of 5 bedding treatments every other day: 0.5 kg of hydrated limestone, 120 mL of commercial acidic conditioner, 1 kg of coal fly ash, 1 kg of kiln-dried wood shavings, and control (no bedding). Counts of coliforms, Klebsiella spp., Escherichia coli, and Streptococcus spp. were lowest on mattresses bedded with lime. Mattresses bedded with the commercial acidic conditioner had the next lowest counts for coliforms, Klebsiella spp., and Streptococcus spp. Wood shavings and the no-bedding control had the highest counts for coliform and Klebsiella spp. Compared with wood shavings or control, fly ash reduced the counts of coliforms, whereas for the other 3 bacterial groups, the reduction was not always significant. Streptococcus spp. counts were greatest in the control group and did not differ among the shavings and fly ash groups. Teat swab results indicated that hydrated lime was the only bedding treatment that significantly decreased the counts of both coliforms and Klebsiella spp. There were no differences in Streptococcus spp. numbers on the teats between any of the bedding treatments. Bacterial populations grew steadily on mattresses and were generally higher at 36 to 48 h than at 12 to 24 h, whereas bacterial populations on teats grew rapidly by 12 h and then remained constant. Hydrated lime was the only treatment that significantly reduced bacterial counts on both mattresses and teat ends, but it caused some skin irritation.

Evaluation of free-stall mattress bedding treatments to reduce mastitis bacterial growth

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

Kristula, M.A.; Dou, Z.; Toth, J.D.

2008-05-15

Bacterial counts were compared in free-stall mattresses and teat ends exposed to 5 treatments in a factorial study design on 1 dairy farm. Mattresses in five 30-cow groups were subjected to 1 of 5 bedding treatments every other day: 0.5 kg of hydrated limestone, 120 mL of commercial acidic conditioner, 1 kg of coal fly ash, 1 kg of kiln-dried wood shavings, and control (no bedding). Counts of coliforms, Klebsiella spp., Escherichia coli, and Streptococcus spp. were lowest on mattresses bedded with lime. Mattresses bedded with the commercial acidic conditioner had the next lowest counts for coliforms, Klebsiella spp., andmore » Streptococcus spp. Wood shavings and the no-bedding control had the highest counts for coliform and Klebsiella spp. Compared with wood shavings or control, fly ash reduced the counts of coliforms, whereas for the other 3 bacterial groups, the reduction was not always significant. Streptococcus spp. counts were greatest in the control group and did not differ among the shavings and fly ash groups. Teat swab results indicated that hydrated lime was the only bedding treatment that significantly decreased the counts of both coliforms and Klebsiella spp. There were no differences in Streptococcus spp. numbers on the teats between any of the bedding treatments. Bacterial populations grew steadily on mattresses and were generally higher at 36 to 48 h than at 12 to 24 h, whereas bacterial populations on teats grew rapidly by 12 h and then remained constant. Hydrated lime was the only treatment that significantly reduced bacterial counts on both mattresses and teat ends, but it caused some skin irritation.« less

Biofilms associated with poultry processing equipment.

PubMed

Lindsay, D; Geornaras, I; von Holy, A

1996-01-01

Aerobic and Gram-negative bacteria were enumerated on non-metallic surfaces and stainless steel test pieces attached to equipment surfaces by swabbing and a mechanical dislodging procedure, respectively, in a South African grade B poultry processing plant. Changes in bacterial numbers were also monitored over time on metal test pieces. The highest bacterial counts were obtained from non-metallic surfaces such as rubber fingered pluckers and plastic defeathering curtains which exceeded the highest counts found on the metal surfaces by at least 1 log CFU cm-2. Gram-negative bacterial counts on all non-metallic surface types were at least 2 log CFU cm-2 lower than corresponding aerobic plate counts. On metal surfaces, the highest microbial numbers were obtained after 14 days exposure, with aerobic plate counts ranging from 3.57 log CFU cm-2 to 5.13 log CFU cm-2, and Gram-negative counts from 0.70 log CFU cm-2 to 3.31 log CFU cm-2. Scanning electron microscopy confirmed the presence of bacterial cells on non-metallic and metallic surfaces associated with poultry processing. Rubber 'fingers', plastic curtains, conveyor belt material and stainless steel test surfaces placed on the scald tank overflow and several chutes revealed extensive and often confluent bacterial biofilms. Extracellular polymeric substances, but few bacterial cells were visible on test pieces placed on evisceration equipment, spinchiller blades and the spinchiller outlet.

Real-time ArcGIS and heterotrophic plate count based chloramine disinfectant control in water distribution system.

PubMed

Bai, Xiaohui; Zhi, Xinghua; Zhu, Huifeng; Meng, Mingqun; Zhang, Mingde

2015-01-01

This study investigates the effect of chloramine residual on bacteria growth and regrowth and the relationship between heterotrophic plate counts (HPCs) and the concentration of chloramine residual in the Shanghai drinking water distribution system (DWDS). In this study, models to control HPCs in the water distribution system and consumer taps are also developed. Real-time ArcGIS was applied to show the distribution and changed results of the chloramine residual concentration in the pipe system by using these models. Residual regression analysis was used to get a reasonable range of the threshold values that allows the chloramine residual to efficiently inhibit bacteria growth in the Shanghai DWDS; the threshold values should be between 0.45 and 0.5 mg/L in pipe water and 0.2 and 0.25 mg/L in tap water. The low residual chloramine value (0.05 mg/L) of the Chinese drinking water quality standard may pose a potential health risk for microorganisms that should be improved. Disinfection by-products (DBPs) were detected, but no health risk was identified.

What's growing on your stethoscope? (And what you can do about it).

PubMed

Schroeder, Ariel; Schroeder, Maryellen A; D'Amico, Frank

2009-08-01

Studies have shown that rubbing alcohol pads on stethoscope diaphragms can reduce bacterial colonization, but alcohol pads are used infrequently used and not always available. We conducted a prospective, single-blinded study to investigate whether simultaneously scrubbing hands and stethoscope head with alcohol-based hand foam would significantly reduce bacterial counts on the stethoscope. Using their own stethoscope, participants imprinted the stethoscope head onto a chocolate agar plate, then used alcohol-based hand foam to cleanse their hands while simultaneously rubbing the stethoscope head. Once the stethoscope heads were dry, the participants imprinted their stethoscope heads onto a second plate. After 48 hours' incubation, we determined the bacterial counts for the prewash and post-wash plates, and compared the 2. We analyzed a total of 184 cultures (from 92 stethoscopes). Both the mean (28 prewash vs 3 post-wash, P=.001) and median (11 prewash vs 1 post-wash, P=.001) colony counts were significantly greater before being cleansed. Three methicillin-resistant Staphylococcus aureus (MRSA) colonies were identified in the prewash period; all were destroyed by the foam. The estimated number of hand washes needed to prevent 1 MRSA colony is 31 (95% confidence interval [CI], 18-89). Simultaneously using hand foam to clean hands and stethoscope heads reduces bacterial counts on stethoscopes. Further research is needed to determine whether this intervention can reduce morbidity and mortality associated with bacterial infection.

Evaluation of procalcitonin and neopterin level in serum of patients with acute bacterial infection.

PubMed

Pourakbari, Babak; Mamishi, Setareh; Zafari, Javid; Khairkhah, Hanieh; Ashtiani, Mohammad H; Abedini, Masomeh; Afsharpaiman, Shahla; Rad, Soroush Seifi

2010-01-01

Fever as a common presenting complaint in pediatric patients can be due to various causes. Differentiating bacterial infection from other causes is important because the prompt use of antibiotics is critical in bacterial infection. Traditional markers of infection such as BT and WBC count may be unspecific and culture may be late or absent. CRP and Procalcitonin (PCT) have been considered to evaluate the evolution of infections and sepsis in patients presenting with SIRS. Neopterin has also been proposed to aid in the diagnosis of bacterial infection. In this study, we compared the value of the serum PCT, neopterin level, and WBC count for predicting bacterial infection and outcome in children with fever. 158 pediatric (2-120-month-old) patients suspected to have acute bacterial infection, based on clinical judgment in which other causes of SIRS were ruled out were included in the study. WBC count with differential was determined and PCT and neopterin levels were measured. PCT level was higher in bacterial infection and patients who were complicated or expired. Rapid PCT test is superior to neopterin and WBC count for anticipating bacterial infection, especially in ED where prompt decision making is critical.

New evidence for Cu-decorated binary-oxides mediating bacterial inactivation/mineralization in aerobic media.

PubMed

Rtimi, S; Pulgarin, C; Bensimon, M; Kiwi, J

2016-08-01

Binary oxide semiconductors TiO2-ZrO2 and Cu-decorated TiO2-ZrO2 (TiO2-ZrO2-Cu) uniform films were sputtered on polyester (PES). These films were irradiated under low intensity solar simulated light and led to bacterial inactivation in aerobic and anaerobic media as evaluated by CFU-plate counting. But bacterial mineralization was only induced by TiO2-ZrO2-Cu in aerobic media. The highly oxidative radicals generated on the films surface under light were identified by the use of appropriate scavengers. The hole generated on the TiO2-ZrO2 films is shown to be the main specie leading to bacterial inactivation. TiO2-ZrO2 and Cu-decorated TiO2-ZrO2 films release Zr and Ti <1ppb and Cu 4.6ppb/cm(2) as determined by inductively coupled plasma mass spectrometry (ICP-MS) This level is far below the citotoxicity permitted level allowed for mammalian cells suggesting that bacterial disinfection proceeds through an oligodynamic effect. By Fourier transform attenuated infrared spectroscopy (ATR-FTIR) the systematic shift of the predominating νs(CH2) vibrational-rotational peak making up most of the bacterial cell-wall content in C was monitored. Based on this evidence a mechanism suggested leading to CH bond stretching followed by cell lysis and cell death. Bacterial inactivation cycling was observed on TiO2-ZrO2-Cu showing the stability of these films leading to bacterial inactivation. Copyright © 2016 Elsevier B.V. All rights reserved.

Fluoro-luminometric real-time measurement of bacterial viability and killing.

PubMed

Lehtinen, Janne; Virta, Marko; Lilius, Esa Matti

2003-10-01

The viability and killing of Escherichia coli was measured on a real-time basis using a fluoro-luminometric device, which allows successive measurements of fluorescence and bioluminescence without user intervention. Bacteria were made fluorescent and bioluminescent by expression of gfp and insect luciferase (lucFF) genes. The green fluorescent protein (GFP) is a highly fluorescent, extremely stable protein, which accumulates in cells during growth, and therefore the measured fluorescence signal was proportional to the total number of cells. The luciferase reaction is dependent of ATP produced by living cells, so that the bioluminescence level was a direct measure of the viable cells. In contrast to the bacterial luciferase, the insect luciferase uses a water-soluble and nonvolatile substrate, which makes automated multi-well microplate assay possible. For the validation of the assay, the proportion of living and dead cell populations was experimentally modified by incubating E. coli cells in the presence of various ethanol concentrations. Bacterial viability and killing measured by a fluoro-luminometric assay correlated fairly well with the reference methods: conventional plate counting, optical density measurement and various flow cytometric analyses. The real-time assay described here allows following the changes in bacterial cultures and assessing the bactericidal and other effects of various chemical, immunological and physical agents simultaneously in large numbers of samples.

Live/Dead Bacterial Spore Assay Using DPA-Triggered Tb Luminescence

NASA Technical Reports Server (NTRS)

Ponce, Adrian

2003-01-01

A method of measuring the fraction of bacterial spores in a sample that remain viable exploits DPA-triggered luminescence of Tb(3+) and is based partly on the same principles as those described earlier. Unlike prior methods for performing such live/dead assays of bacterial spores, this method does not involve counting colonies formed by cultivation (which can take days), or counting of spores under a microscope, and works whether or not bacterial spores are attached to other small particles (i.e., dust), and can be implemented on a time scale of about 20 minutes.

Effect of Manuka honey gel on the transforming growth factor β1 and β3 concentrations, bacterial counts and histomorphology of contaminated full-thickness skin wounds in equine distal limbs.

PubMed

Bischofberger, A S; Dart, C M; Horadagoda, N; Perkins, N R; Jeffcott, L B; Little, C B; Dart, A J

2016-01-01

To investigate the effect of 66% Manuka honey gel on the concentrations of transforming growth factor (TGF)-β1 and TGF-β3, bacterial counts and histomorphology during healing of contaminated equine distal limb wounds. In this experimental study of 10 Standardbred horses, five full-thickness skin wounds (2 × 1.5 cm) were created on one metacarpus and six similar wounds were created on the contralateral metacarpus. Wounds were assigned to three groups: non-contaminated control wounds; contaminated control wounds; contaminated wounds treated daily with 1 mL Manuka honey gel topically for 10 days. For the contaminated wounds, faeces were applied for 24 h after wound creation. In five horses wounds were bandaged and in the other five horses wounds were left without a bandage. Biopsies were taken on days 1, 2, 7 and 10 after wounding to evaluate the effects of Manuka honey gel, wound contamination and bandaging on TGF-β1 and TGF-β3 concentrations, aerobic and anaerobic bacterial counts, and histomorphology. Manuka honey gel had no significant effect on TGF-β1 and TGF-β3 concentrations or wound bacterial counts. Manuka honey gel decreased wound inflammation (days 7, 10), increased angiogenesis (days 2, 7, 10), increased fibrosis and collagen organisation (day 7) and increased epithelial hyperplasia (days 7, 10). Treatment with Manuka honey gel resulted in a more organised granulation tissue bed early in wound repair, which may contribute to enhanced healing of equine distal limb wounds. © 2016 Australian Veterinary Association.

Rapid staining and enumeration of small numbers of total bacteria in water by solid-phase laser cytometry

NASA Technical Reports Server (NTRS)

Broadaway, Susan C.; Barton, Stephanie A.; Pyle, Barry H.

2003-01-01

The nucleic acid stain SYBR Green I was evaluated for use with solid-phase laser cytometry to obtain total bacterial cell counts from several water sources with small bacterial numbers. Results were obtained within 30 min and exceeded or equaled counts on R2A agar plates incubated for 14 days at room temperature.

Ultraviolet micro-Raman spectrograph for the detection of small numbers of bacterial cells

NASA Astrophysics Data System (ADS)

Chadha, S.; Nelson, W. H.; Sperry, J. F.

1993-11-01

The construction of a practical UV micro-Raman spectrograph capable of selective excitation of bacterial cells and other microscopic samples has been described. A reflective objective is used to focus cw laser light on a sample and at the same time collect the scattered light at 180°. With the aid of a quartz lens the image produced is focused on the slits of a spectrograph equipped with a single 2400 grooves/mm grating optimized for 250 nm. Spectra were detected by means of a blue-intensified diode array detector. Resonance Raman spectra of Bacillus subtilis and Flavobacterium capsulatum excited by the 257.2 nm output of a cw laser were recorded in the 900-1800 cm-1 region. Bacterial cells were immobilized on a quartz plate by means of polylysine and were counted visually. Cooling was required to retard sample degradation. Sample sizes ranged from 1 to 50 cells with excitation times varying from 15 to 180 s. Excellent spectra have been obtained from 20 cells in 15 s using a spectrograph having only 3% throughput.

The importance of the viable but non-culturable state in human bacterial pathogens

PubMed Central

Li, Laam; Mendis, Nilmini; Trigui, Hana; Oliver, James D.; Faucher, Sebastien P.

2014-01-01

Many bacterial species have been found to exist in a viable but non-culturable (VBNC) state since its discovery in 1982. VBNC cells are characterized by a loss of culturability on routine agar, which impairs their detection by conventional plate count techniques. This leads to an underestimation of total viable cells in environmental or clinical samples, and thus poses a risk to public health. In this review, we present recent findings on the VBNC state of human bacterial pathogens. The characteristics of VBNC cells, including the similarities and differences to viable, culturable cells and dead cells, and different detection methods are discussed. Exposure to various stresses can induce the VBNC state, and VBNC cells may be resuscitated back to culturable cells under suitable stimuli. The conditions that trigger the induction of the VBNC state and resuscitation from it are summarized and the mechanisms underlying these two processes are discussed. Last but not least, the significance of VBNC cells and their potential influence on human health are also reviewed. PMID:24917854

Impact of solar radiation exposure on phyllosphere bacterial community of red-pigmented baby leaf lettuce.

PubMed

Truchado, Pilar; Gil, M Isabel; Reboleiro, Patricia; Rodelas, Belén; Allende, Ana

2017-09-01

Solar radiation has been identified as a stress factor affecting phyllosphere associated bacteria colonization and survival during primary production. In the present study, the impact of different solar radiation doses on the phyllosphere microbiota of red-pigmented baby leaf lettuce cultivated in open field under commercial conditions was evaluated. Four weeks before harvest, the growing field was divided into four plots; each one was consecutively covered with one-week-interval with a light-excluding plastic to reduce the sunlight exposure. Four different solar radiation treatments were generated and cumulative photosynthetically active radiation (PAR) was used to differentiate treatments as follows: 4889 ± 428 μmol/m 2 /s (uncovered), 4265 ± 356 μmol/m 2 /s (covered for 1 week), 3602 ± 225 μmol/m 2 /s (covered for 2 weeks) and 3115 ± 313 μmol/m 2 /s (covered for 3 weeks). The size and composition of the phyllosphere bacterial community were determined by cultivation-depended (plate count) and independent (qPCR) techniques. Exposure to decreased levels of cumulative PAR did not produce significant differences in total bacterial community size, regardless of the chosen quantification techniques. However, total bacteria size quantified by qPCR was around 3.5 orders of magnitude higher than those obtained by plate count. The observed differences between cultivation-depended and independent techniques could be attributed to the presence of non-viable or viable but non-culturable (VBNC) bacteria. The bacterial community structure was analyzed using temperature gradient gel electrophoresis (TGGE), and significant differences were detected when the four solar treatment were compared. A qPCR approach was applied to the quantification of specific bacterial phyla and classes, previously identified in the phyllosphere of plants available literature, confirming that Proteobacteria, Bacteroidetes, Actinobacterias and Firmicutes were the most abundantly

Characterization of bacterial communities and functions of two submerged soils from San Vitale park (Italy)

NASA Astrophysics Data System (ADS)

Mocali, Stefano; Chiellini, Carolina; Lagomarsino, Alessandra; Ferronato, Chiara; Vittori Antisari, Livia; Vianello, Gilmo

2015-04-01

Subaqueous soils has been introduced in the last edition of the Keys to Soil Taxonomy (Soil surveystaff, 2014), to describe soils covered by a water column of up to 2.5 m where different pedogenetic processes can be recognized. However, the role of bacterial community structure and function in such environments and its potential use as pedogenetic indicator is still largely unknown. Two submerged soils (WAS-2 and WAS-4) were collected from San Vitale park (Italy), a site where the evolution of the landscape from subaqueous wetland to interdunal and dunal system, and the interfacing of freshwater with saltwater, made this site particularly suitable for examining the pedogenetic indicators which can characterize and predict the soil hydromorphism in trasitional ecosystems. The two soils were classified and their physicochemical and morphological features were investigated. Selective media were used to isolate both culturable aerobic and anaerobic (microaerophilic) bacteria associated with each horizon. In WAS-2 seven horizons were identified (depths 4-0, 0-6, 6-13, 13-20, 20-36, 36-59/60, and 59/60-83 cm) while in WAS-4, five horizons were identified (depths 0-14, 14-20, 20-40, 40-45, 45-100 cm) for a total of 12 horizons (samples). For each sample, aerobic bacterial plate count was performed on solid LB medium, coupled with microaerophilic bacterial plate count either on SA500 minimal medium and AYE medium (0.5% soft agar each). Molecular identification (16S rRNA gene sequencing) of ~100 strains isolated from each of the three used medium was performed, for a total of ~300 strains for each sample. To complete the characterization of the microbial communities in all horizons, Next Generation Sequencing (NGS) analysis was carried out with 454 platform on each of the 12 samples. Moreover, the N2O and CH4 emissions were determined from each pedon. All the parameters were used to highlight the similarities and the differences between and within the pedons. The results

Effect of surgical hand scrub time on subsequent bacterial growth.

PubMed

Wheelock, S M; Lookinland, S

1997-06-01

In this experimental study, the researchers evaluated the effect of surgical hand scrub time on subsequent bacterial growth and assessed the effectiveness of the glove juice technique in a clinical setting. In a randomized crossover design, 25 perioperative staff members scrubbed for two or three minutes in the first trial and vice versa in the second trial, after which the wore sterile surgical gloves for one hour under clinical conditions. The researchers then sampled the subjects' nondominant hands for bacterial growth, cultured aliquots from the sampling solution, and counted microorganisms. Scrubbing for three minutes produced lower mean log bacterial counts than scrubbing for two minutes. Although the mean bacterial count differed significantly (P = .02) between the two-minute and three-minute surgical hand scrub times, it fell below 0.5 log, which is the threshold for practical and clinical significance. This finding suggests that a two-minute surgical hand scrub is clinically as effective as a three-minute surgical had scrub. The glove juice technique demonstrated sensitivity and reliability in enumerating bacteria on the hands of perioperative staff members in a clinical setting.

In vitro assessments on bacterial adhesion and corrosion performance of TiN coating on Ti6Al4V titanium alloy synthesized by multi-arc ion plating

NASA Astrophysics Data System (ADS)

Lin, Naiming; Huang, Xiaobo; Zhang, Xiangyu; Fan, Ailan; Qin, Lin; Tang, Bin

2012-07-01

TiN coating was synthesized on Ti6Al4V titanium alloy surface by multi-arc ion plating (MIP) technique. Surface morphology, cross sectional microstructure, elemental distributions and phase compositions of the obtained coating were analyzed by means of scanning electron microscope (SEM), optical microscope (OM), glow discharge optical emission spectroscope (GDOES) and X-ray diffraction (XRD). Bacterial adhesion and corrosion performance of Ti6Al4V and the TiN coating were assessed via in vitro bacterial adhesion tests and corrosion experiments, respectively. The results indicated that continuous and compact coating which was built up by pure TiN with a typical columnar crystal structure has reached a thickness of 1.5 μm. This TiN coating could significantly reduce the bacterial adhesion and enhance the corrosion resistance of Ti6Al4V substrate.

Inactivation of selected bacterial pathogens in dairy cattle manure by mesophilic anaerobic digestion (balloon type digester).

PubMed

Manyi-Loh, Christy E; Mamphweli, Sampson N; Meyer, Edson L; Okoh, Anthony I; Makaka, Golden; Simon, Michael

2014-07-14

Anaerobic digestion of animal manure in biogas digesters has shown promise as a technology in reducing the microbial load to safe and recommended levels. We sought to treat dairy manure obtained from the Fort Hare Dairy Farm by investigating the survival rates of bacterial pathogens, through a total viable plate count method, before, during and after mesophilic anaerobic digestion. Different microbiological media were inoculated with different serial dilutions of manure samples that were withdrawn from the biogas digester at 3, 7 and 14 day intervals to determine the viable cells. Data obtained indicated that the pathogens of public health importance were 90%-99% reduced in the order: Campylobacter sp. (18 days) < Escherichia coli sp. (62 days) < Salmonella sp. (133 days) from a viable count of 10.1 × 103, 3.6 × 105, 7.4 × 103 to concentrations below the detection limit (DL = 102 cfu/g manure), respectively. This disparity in survival rates may be influenced by the inherent characteristics of these bacteria, available nutrients as well as the stages of the anaerobic digestion process. In addition, the highest p-value i.e., 0.957 for E. coli showed the statistical significance of its model and the strongest correlation between its reductions with days of digestion. In conclusion, the results demonstrated that the specific bacterial pathogens in manure can be considerably reduced through anaerobic digestion after 133 days.

Short communication: Environmental mastitis pathogen counts in freestalls bedded with composted and fresh recycled manure solids.

PubMed

Cole, K J; Hogan, J S

2016-02-01

An experiment was conducted to compare bacterial counts of environmental mastitis pathogens in composted recycled manure solids bedding with those in fresh recycled manure solids. Eighteen Holstein cows were housed in 1 pen with 18 stalls. One row of 9 freestalls included mattresses and was bedded weekly with composted recycled manure solids. The second row of 9 freestalls included mattresses and was bedded weekly with fresh recycled manure solids. The back one-third of stalls toward the alleyway was covered in 25 to 50 mm of bedding. Samples were taken from the back one-third of 4 stalls for both treatments on d 0, 1, 2, and 6 of each week. After 3 wk, bedding treatments were switched between rows, making the total duration 6 wk. Mean total gram-negative bacterial counts were approximately 0.5 log10 cfu/g of dry matter lower in the composted recycled manure solids on d 0 compared with fresh recycled manure solids. Klebsiella species, coliform, and Streptococcus species counts were at least 1.0 log10 cfu/g of dry matter lower in composted compared with fresh recycled manure solids on d 0. Only gram-negative bacterial counts on d 1 were reduced in composted recycled manure solids compared with fresh recycled manure solids. Differences were not observed between treatments in gram-negative bacterial, coliform, Klebsiella species, or Streptococcus species counts on d 2 and 6. Ash content was higher in composted recycled manure solids compared with fresh recycled manure solids on d 0, 1, 2, and 6. Despite the increase in ash after composting, bacterial counts of mastitis pathogens in composted recycled manure solids were comparable with those in fresh recycled manure when used as freestall bedding. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Radiolabel ratio method for measuring pulmonary clearance of intratracheal bacterial challenges

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

LaForce, F.M.; Boose, D.S.

Calculation of bacterial clearance is a fundamental step in any study of in situ lung antibacterial defenses. A method is described whereby about 85% of a radiolabeled bacterial inoculum was consistently introduced into the bronchopulmonary tree of a mouse by the intratracheal route. Mice were then killed 1 and 4 hours later; their lungs were removed aseptically and homogenized, and viable bacteria and radiolabel counts were determined. Radiolabel counts fell slowly, and more than 80% of the original radiolabel was still present in homogenized lung samples from animals sacrificed 4 hours after challenge. Bacteria/isotope ratios for the bacterial inoculum andmore » homogenized lung samples from animals sacrificed immediately after challenge were very similar. Bacterial clearance values were the same whether computed from bacterial counts alone or according to a radiolabel ratio method whereby the change in the bacteria/isotope ratio in ground lung aliquots was divided by a similar ratio from bacteria used to inoculate animals. Some contamination resulted from oral streptococci being swept into the bronchopulmonary free during the aspiration process. This contamination was not a problem when penicillin was incorporated into the agar and penicillin-resistant strains were used for the bacterial challenges.« less

Do bacterial cell numbers follow a theoretical Poisson distribution? Comparison of experimentally obtained numbers of single cells with random number generation via computer simulation.

PubMed

Koyama, Kento; Hokunan, Hidekazu; Hasegawa, Mayumi; Kawamura, Shuso; Koseki, Shigenobu

2016-12-01

We investigated a bacterial sample preparation procedure for single-cell studies. In the present study, we examined whether single bacterial cells obtained via 10-fold dilution followed a theoretical Poisson distribution. Four serotypes of Salmonella enterica, three serotypes of enterohaemorrhagic Escherichia coli and one serotype of Listeria monocytogenes were used as sample bacteria. An inoculum of each serotype was prepared via a 10-fold dilution series to obtain bacterial cell counts with mean values of one or two. To determine whether the experimentally obtained bacterial cell counts follow a theoretical Poisson distribution, a likelihood ratio test between the experimentally obtained cell counts and Poisson distribution which parameter estimated by maximum likelihood estimation (MLE) was conducted. The bacterial cell counts of each serotype sufficiently followed a Poisson distribution. Furthermore, to examine the validity of the parameters of Poisson distribution from experimentally obtained bacterial cell counts, we compared these with the parameters of a Poisson distribution that were estimated using random number generation via computer simulation. The Poisson distribution parameters experimentally obtained from bacterial cell counts were within the range of the parameters estimated using a computer simulation. These results demonstrate that the bacterial cell counts of each serotype obtained via 10-fold dilution followed a Poisson distribution. The fact that the frequency of bacterial cell counts follows a Poisson distribution at low number would be applied to some single-cell studies with a few bacterial cells. In particular, the procedure presented in this study enables us to develop an inactivation model at the single-cell level that can estimate the variability of survival bacterial numbers during the bacterial death process. Copyright © 2016 Elsevier Ltd. All rights reserved.

Bacterial transport in heterogeneous porous media: Observations from laboratory experiments

NASA Astrophysics Data System (ADS)

Silliman, S. E.; Dunlap, R.; Fletcher, M.; Schneegurt, M. A.

2001-11-01

Transport of bacteria through heterogeneous porous media was investigated in small-scale columns packed with sand and in a tank designed to allow the hydraulic conductivity to vary as a two-dimensional, lognormally distributed, second-order stationary, exponentially correlated random field. The bacteria were Pseudomonas ftuorescens R8, a strain demonstrating appreciable attachment to surfaces, and strain Ml, a transposon mutant of strain R8 with reduced attachment ability. In bench top, sand-filled columns, transport was determined by measuring intensity of fluorescence of stained cells in the effluent or by measuring radiolabeled cells that were retained in the sand columns. Results demonstrated that strain Ml was transported more efficiently than strain R8 through columns packed with either a homogeneous silica sand or a more heterogeneous sand with iron oxide coatings. Two experiments conducted in the tank involved monitoring transport of bacteria to wells via sampling from wells and sample ports in the tank. Bacterial numbers were determined by direct plate count. At the end of the first experiment, the distribution of the bacteria in the sediment was determined by destructive sampling and plating. The two experiments produced bacterial breakthrough curves that were quite similar even though the similarity between the two porous media was limited to first- and second-order statistical moments. This result appears consistent with the concept of large-scale, average behavior such as has been observed for the transport of conservative chemical tracers. The transported bacteria arrived simultaneously with a conservative chemical tracer (although at significantly lower normalized concentration than the tracer). However, the bacterial breakthrough curves showed significant late time tailing. The concentrations of bacteria attached to the sediment surfaces showed considerably more spatial variation than did the concentrations of bacteria in the fluid phase. This

Single electron counting using a dual MCP assembly

NASA Astrophysics Data System (ADS)

Yang, Yuzhen; Liu, Shulin; Zhao, Tianchi; Yan, Baojun; Wang, Peiliang; Yu, Yang; Lei, Xiangcui; Yang, Luping; Wen, Kaile; Qi, Ming; Heng, Yuekun

2016-09-01

The gain, pulse height resolution and peak-to-valley ratio of single electrons detected by using a Chevron configured Microchannel Plate (MCP) assembly are studied. The two MCPs are separated by a 280 μm gap and are biased by four electrodes. The purpose of the study is to determine the optimum bias voltage arrangements for single electron counting. By comparing the results of various bias voltage combinations, we conclude that good performance for the electron counting can be achieved by operating the MCP assembly in saturation mode. In addition, by applying a small reverse bias voltage across the gap while adjusting the bias voltages of the MCPs, optimum performance of electron counting can be obtained.

An investigation of total bacterial communities, culturable antibiotic-resistant bacterial communities and integrons in the river water environments of Taipei city.

PubMed

Yang, Chu-Wen; Chang, Yi-Tang; Chao, Wei-Liang; Shiung, Iau-Iun; Lin, Han-Sheng; Chen, Hsuan; Ho, Szu-Han; Lu, Min-Jheng; Lee, Pin-Hsuan; Fan, Shao-Ning

2014-07-30

The intensive use of antibiotics may accelerate the development of antibiotic-resistant bacteria (ARB). The global geographical distribution of environmental ARB has been indicated by many studies. However, the ARB in the water environments of Taiwan has not been extensively investigated. The objective of this study was to investigate the communities of ARB in Huanghsi Stream, which presents a natural acidic (pH 4) water environment. Waishuanghsi Stream provides a neutral (pH 7) water environment and was thus also monitored to allow comparison. The plate counts of culturable bacteria in eight antibiotics indicate that the numbers of culturable carbenicillin- and vancomycin-resistant bacteria in both Huanghsi and Waishuanghsi Streams are greater than the numbers of culturable bacteria resistant to the other antibiotics tested. Using a 16S rDNA sequencing approach, both the antibiotic-resistant bacterial communities (culture-based) and the total bacterial communities (metagenome-based) in Waishuanghsi Stream exhibit a higher diversity than those in Huanghsi Stream were observed. Of the three classes of integron, only class I integrons were identified in Waishuanghsi Stream. Our results suggest that an acidic (pH 4) water environment may not only affect the community composition of antibiotic-resistant bacteria but also the horizontal gene transfer mediated by integrons. Copyright © 2013 Elsevier B.V. All rights reserved.

Influence of pre- and post-usage flushing frequencies on bacterial water quality of non-touch water fittings.

PubMed

Suchomel, Miranda; Diab-Elschahawi, Magda; Kundi, Michael; Assadian, Ojan

2013-08-30

Non-touch fittings have been reported to be susceptible for Pseudomonas aeruginosa accumulation. A number of factors may contribute to this, including the frequency of usage, duration of water stagnation, or presence of plastic materials. Programmable non-touch fittings are appearing which allow regular automated post-flushing with cold water to prevent water stagnation. However, the ideal duration of post-flushing is unknown as well as the effect of pre-rinsing with cold water before use. Eight non-touch fittings with brass valve blocks were mounted on a mobile test sink and connected to the same central water pipe source, differing only in presence or absence of water connection pipes, length of connection pipe, frequency of usage, and time intervals for pre- and post-usage water flush. The total bacteria colony-forming unit (cfu) counts were obtained by the spread plate technique. Low frequency of water use in combination with a long stagnating water column resulted in high bacterial cfu counts. Post-usage flushing for 2 seconds did not differ from no flushing. Flushing for 10 seconds with cold water after use or 30 seconds flush before use were both the most effective measures to prevent non-touch fittings from biofilm formation over a period of 20 weeks. Further improvements in water fitting technology could possibly solve the problem of bacterial water contamination in health care settings.

Effects of Bite Count Feedback from a Wearable Device and Goal Setting on Consumption in Young Adults.

PubMed

Jasper, Phillip W; James, Melva T; Hoover, Adam W; Muth, Eric R

2016-11-01

New technologies are emerging that may help individuals engage in healthier eating behaviors. One paradigm to test the efficacy of a technology is to determine its effect relative to environment cues that are known to cause individuals to overeat. The purpose of this work was to independently investigate two questions: How does the presence of a technology that provides bite count feedback alter eating behavior? and, How does the presence of a technology that provides bite count feedback paired with a goal alter eating behavior? Two studies investigated these research questions. The first study tested the effects of a large and small plate crossed with the presence or absence of a device that provided bite count feedback on intake. The second study tested the effects of a bite count goal with bite count feedback, again crossed with plate size, on intake. Both studies used a 2×2 between-subjects design. In the first study, 94 subjects (62 women aged 19.0±1.6 years with body mass index [BMI] 23.04±3.6) consumed lunch in a laboratory. The second study examined 99 subjects (56 women aged 18.5±1.5 years with BMI 22.73±2.70) under the same conditions. In both studies subjects consumed a single-course meal, using either a small or large plate. In the first study participants either wore or did not wear an automated bite counting device. In the second study all participants wore the bite counting device and were given either a low bite count goal (12 bites) or a high bite count goal (22 bites). Effect of plate size, feedback, and goal on consumption (grams) and number of bites taken were assessed using 2×2 analyses of variance. As adjunct measures, the effects of serving size, bite size (grams per bite), postmeal satiety, and satiety change were also assessed. In the first study there was a main effect of plate size on grams consumed and number of bites taken such that eating from a large plate led to greater consumption (P=0.001) and a greater number of bites (P=0

Cook/chill foodservice system with a microwave oven: aerobic plate counts from beef loaf, potatoes and frozen green beans.

PubMed

Dahl, C A; Matthews, M E; Marth, E H

1980-06-01

The purpose was to evaluate microbiological quality and end temperature (ET) of portioned food after heating in a microwave oven as used in a hospital cook/chill foodservice system. Beef loaf (15 kg), potatoes (6 kg), and green beans (5 kg) were prepared in a laboratory. After initial cooking to 60 degrees C, and storage (7 degrees C for 24 h), beef loaf (100 g) was microwave heated: 20, 50, 80 or 110 s. Potatoes were reconstituted, stored (7 degrees C for 24 h), portioned (100 g/portion), and microwave-heated: 25, 45, 65 or 84 s. Beans were thawed (7 degrees C for 24 h), portioned (100 g/portion), and microwave-heated: 20, 50, 80 or 110 s. Aerobic plate counts (APC) for foods were obtained throughout product flow. Wide ranges of Et and of APC in foods indicates that research is needed, for greater control of microwave-heating through advanced microwave engineering and food technology, to produce food with constant microbiological quality.

An anti-bacterial approach to nanoscale roughening of biomimetic rice-like pattern PP by thermal annealing

NASA Astrophysics Data System (ADS)

Jafari Nodoushan, Emad; Ebrahimi, Nadereh Golshan; Ayazi, Masoumeh

2017-11-01

In this paper, we introduced thermal annealing treatment as an effective way of increasing the nanoscale roughness of a semi-crystalline polymer surface. Annealing treatment applied to a biomimetic microscale pattern of rice leaf to achieve a superhydrophobic surface with a hierarchical roughness. Resulted surfaces was characterized by XRD, AFM and FE-SEM instruments and showed an increase of roughness and cristallinity within both time and temperature of treatment. These two parameters also impact on measured static contact angle up to 158°. Bacterial attachment potency has an inverse relationship with the similarity of surface pattern dimensions and bacterial size and due to that, thermal annealing could be an effective way to create anti-bacterial surface beyond its effect on water repellency. Point in case, the anti-bacterial properties of produced water-repellence surfaces of PP were measured and counted colonies of both gram-negative (E. coli) and gram-positive (S. aureus) bacteria reduced with the nature of PP and hierarchical pattern on that. Anti-bacterial characterization of the resulted surface reveals a stunning reduction in adhesion of gram-positive bacteria to the surface. S. aureus reduction rates equaled to 95% and 66% when compared to control blank plate and smooth surface of PP. Moreover, it also could affect the other type of bacteria, gram-negative (E. coli). In the latter case, adhesion reduction rates calculated 66% and 53% when against to the same controls, respectively.

Hydrodynamics of bacterial colonies: Phase diagrams

NASA Astrophysics Data System (ADS)

Lega, J.; Passot, T.

2004-09-01

We present numerical simulations of a recent hydrodynamic model describing the growth of bacterial colonies on agar plates. We show that this model is able to qualitatively reproduce experimentally observed phase diagrams, which relate a colony shape to the initial quantity of nutrients on the plate and the initial wetness of the agar. We also discuss the principal features resulting from the interplay between hydrodynamic motions and colony growth, as described by our model.

Comparison of Dry Medium Culture Plates for Mesophilic Aerobic Bacteria in Milk, Ice Cream, Ham, and Codfish Fillet Products

PubMed Central

Park, Junghyun; Kim, Myunghee

2013-01-01

This study was performed to compare the performance of Sanita-Kun dry medium culture plate with those of traditional culture medium and Petrifilm dry medium culture plate for the enumeration of the mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet. Mesophilic aerobic bacteria were comparatively evaluated in milk, ice cream, ham, and codfish fillet using Sanita-Kun aerobic count (SAC), Petrifilm aerobic count (PAC), and traditional plate count agar (PCA) media. According to the results, all methods showed high correlations of 0.989~1.000 and no significant differences were observed for enumerating the mesophilic aerobic bacteria in the tested food products. SAC method was easier to perform and count colonies efficiently as compared to the PCA and PAC methods. Therefore, we concluded that the SAC method offers an acceptable alternative to the PCA and PAC methods for counting the mesophilic aerobic bacteria in milk, ice cream, ham, and codfish fillet products. PMID:24551829

RAPA: a novel in vitro method to evaluate anti-bacterial skin cleansing products.

PubMed

Ansari, S A; Gafur, R B; Jones, K; Espada, L A; Polefka, T G

2010-04-01

Development of efficacious anti-bacterial skin cleansing products has been limited by the availability of a pre-clinical (in vitro) method to predict clinical efficacy adequately. We report a simple and rapid method, designated as rapid agar plate assay (RAPA), that uses the bacteriological agar surface as a surrogate substrate for skin and combines elements of two widely used in vivo (clinical) methods (Agar Patch and Cup Scrub). To simulate the washing of the human hand or forearm skin with the test product, trypticase soy agar plates were directly washed with the test product and rinsed under running tap water. After air-drying the washed plates, test bacteria (Staphylococcus aureus or Escherichia coli) were applied and the plates were incubated at 37 degrees C for 18-24 h. Using S. aureus as the test organism, anti-bacterial bar soap containing triclocarbanilide showed a strong linear relationship (R(2) = 0.97) between bacterial dose and their per cent reduction. A similar dose-response relationship (R(2) = 0.96) was observed for anti-bacterial liquid hand soap against E. coli. RAPA was able to distinguish between anti-bacterial products based on the nature and level of actives in them. In limited comparative tests, results obtained by RAPA were comparable with the results obtained by clinical agar patch and clinical cup scrub methods. In conclusion, RAPA provides a simple, rugged and reproducible in vitro method for testing the relative efficacy of anti-bacterial skin cleansing products with a likelihood of comparable clinical efficacy. Further testing is warranted to improve the clinical predictability of this method.

[Validation of a clinical prediction rule to distinguish bacterial from aseptic meningitis].

PubMed

Agüero, Gonzalo; Davenport, María C; Del Valle, María de la P; Gallegos, Paulina; Kannemann, Ana L; Bokser, Vivian; Ferrero, Fernando

2010-02-01

Despite most meningitis are not bacterial, antibiotics are usually administered on admission because bacterial meningitis is difficult to be rule-out. Distinguishing bacterial from aseptic meningitis on admission could avoid inappropriate antibiotic use and hospitalization. We aimed to validate a clinical prediction rule to distinguish bacterial from aseptic meningitis in children, on arriving to the emergency room. This prospective study included patients aged < 19 years with meningitis. Cerebrospinal fluid (CSF) and peripheral blood neutrophil count were obtained from all patients. The BMS (Bacterial Meningitis Score) described by Nigrovic (Pediatrics 2002; 110: 712), was calculated: positive CSF Gram stain= 2 points, CSF absolute neutrophil count > or = 1000 cells/mm(3), CSF protein > or = 80 mg/dl, peripheral blood absolute neutrophil count > or = 10.000/mm(3), seizure = 1 point each. Sensitivity (S), specificity (E), positive and negative predictive values (PPV and NPV), positive and negative likelihood ratios (PLR and NLR) of the BMS to predict bacterial meningitis were calculated. Seventy patients with meningitis were included (14 bacterial meningitis). When BMS was calculated, 25 patients showed a BMS= 0 points, 11 BMS= 1 point, and 34 BMS > or = 2 points. A BMS = 0 showed S: 100%, E: 44%, VPP: 31%, VPN: 100%, RVP: 1,81 RVN: 0. A BMS > or = 2 predicted bacterial meningitis with S: 100%, E: 64%, VPP: 41%, VPN: 100%, PLR: 2.8, NLR:0. Using BMS was simple, and allowed identifying children with very low risk of bacterial meningitis. It could be a useful tool to assist clinical decision making.

Bacterial contamination of the hands of food handlers as indicator of hand washing efficacy in some convenient food industries in South Africa.

PubMed

Aa, Lambrechts; Is, Human; Jh, Doughari; Jfr, Lues

2014-07-01

Hands of ready-to-eat food service employees have been shown to be vectors in the spread of foodborne disease, mainly because of poor personal hygiene and accounting for approximately 97% of food borne illnesses in food service establishments and homes. Our objective was to evaluate the efficacy of hand washing practices and sanitation before commencing work among food handlers in the convenient food industry in Gauteng, South Africa. A total of 230 samples were collected, involving 100% of the food handlers, in 8 selected convenient food outlets with their main focus on preparing ready-to-eat foods. The workers' cleaned and disinfected dominant hands were sampled for Total Plate Count (TPC), Staphylococcus aureus and Escherichia coli. Bacteria were isolated and counted using standard methods. The highest bacterial count from the hand samples was 7.4 x 10(3) cfu.cm(-2) and the lowest showed no detectable growth. Although hands with a count of 0 cfu.cm(-2) were found in all of the plants, the results indicated that all the plants exceeded the legal limit for food surfaces or hands of < 100 cfu.cm(-2) when the average bacterial counts on hands were compared. Sixty percent of the TPC analysed exceeded the legal limit and only 18% of the food handlers had no bacteria detectable on their hands. One sample tested positive for E. coli and S. aureus could not be detected on the hands of any of the food handlers. The study revealed that hand hygiene is unsatisfactory and may have serious implications for public health due to contamination of food from food handlers' hands. This therefore underlined the importance of further training to improve food handlers' knowledge of good hand washing practices.

A comparative study of biofilm formation by Shiga toxigenic Escherichia coli using epifluorescence microscopy on stainless steel and a microtitre plate method.

PubMed

Rivas, Lucia; Dykes, Gary A; Fegan, Narelle

2007-04-01

Attachment of Shiga toxigenic Escherichia coli (STEC) to surfaces and the formation of biofilms may enhance persistence in a food processing environment and present a risk of contaminating products. Seven strains of STEC and three non-STEC strains were selected to compare two biofilm quantification methods; epifluorescence microscopy on stainless steel (SS) and a microtitre plate assay. The influence of prior growth in planktonic (nutrient broth) and sessile (nutrient agar) culture on biofilm production, as well as expression of surface structures and the possession of antigen 43 (encoded by agn43) on biofilm formation were also investigated. Biofilms were produced in diluted nutrient broth at 25 degrees C for 24 and 48 h. Curli expression was determined using congo red indicator agar, while the presence of agn43 was determined using polymerase chain reaction. No correlation was found between counts for epifluorescence microscopy on SS and the absorbance values obtained with the microtitre plate method for planktonic and sessile grown cultures. Different abilities of individual STEC strains to attach to SS and microtitre plates were found with some strains attaching better to each surface following growth in either planktonic or sessile culture. All O157 STEC strains had low biofilm counts on SS for planktonic and sessile grown cultures; however, one STEC O157:H- strain (EC516) had significantly greater (p<0.05) biofilm production on microtitre plates compared to the other O157 STEC strains. EC516 and other STEC (O174:H21 and O91:H21) strains expressing curli fimbriae were found to produce significantly greater (p<0.05) biofilms on microtitre plates compared to the non-curli expressing strains. No relationship was found between the production of type-I fimbriae, motility, agn43 and bacterial physicochemical properties (previously determined) and biofilm formation on SS or microtitre plates. Variations between the two biofilm determination methods may suggest that

Aerobic plate counts and ATP levels correlate with Listeria monocytogenes detection in retail delis.

PubMed

Hammons, Susan R; Stasiewicz, Matthew J; Roof, Sherry; Oliver, Haley F

2015-04-01

Listeria monocytogenes is a foodborne pathogen that causes an estimated 1,591 cases of illness and 255 deaths annually in the United States, the majority of which are attributed to ready-to-eat deli meats processed in retail delis. Because retail delis distribute product directly to consumers, rapid methods to validate cleaning and sanitation are needed to improve retail food safety. This study investigated the relationships among ATP levels, standard aerobic plate count (APC), and L. monocytogenes presence in fully operational delis. Fifteen full-service delis were concurrently sampled for ATP, APC, and L. monocytogenes during preoperational hours once monthly for 3 months. Fifteen additional delis were recruited for 6 months of operational sampling (n = 30). A 1-log increase in APC was equivalent to a 3.3-fold increase in the odds of detecting L. monocytogenes (P < 0.001) and a 1.9-log increase in L monocytogenes population (P = 0.03). An ATP level increase of 1 log relative light unit correlated to a 0.22-log increase in APC (P < 0.001). A preoperational ATP level mean increase by 1 log relative light unit increased the odds of detecting L. monocytogenes concurrently fourfold. A 0.5-log increase in mean ATP level during preoperational sampling corresponded to a 2% increase in the predicted L. monocytogenes prevalence during operation (P < 0.01). Additionally, 10 statistically representative sites were identified and recommended for use in sanitation monitoring programs. Our data support the use of ATP as a rapid method to validate effective cleaning and sanitation to reduce L. monocytogenes in retail delis.

Characterization of the dominant bacterial communities during storage of Norway lobster and Norway lobster tails (Nephrops norvegicus) based on 16S rDNA analysis by PCR-DGGE.

PubMed

Bekaert, Karen; Devriese, Lisa; Maes, Sara; Robbens, Johan

2015-04-01

The aim of this study was to investigate the microbial quality of whole Norway lobster (Nephrops norvegicus) and Norway lobster tails to optimize handling conditions. This was done by assessing the total viable count (TVC) and characterizing the dominant microbiota. The cultivable microorganisms were quantified via classical microbiological plating methods. To characterize as many bacterial species present as possible, we performed advanced molecular identification techniques (PCR-DGGE). The initial TVC of fresh Norway lobster meat was high (3.0 log cfu/g) as compared to fish. No significant difference between whole Norway lobster and Norway lobster tails could be found during the storage period. From day 6 of storage, a significant difference between Plate Count Agar (PCA) and Marine Agar (MA) was observed. The microbiota of Norway lobster was dominated by members of the Gram-negative genera such as Psychrobacter spp., Pseudoalteromonas spp., Pseudomonas spp., Luteimonas spp., and Aliivibrio spp. From these bacteria, mainly Psychrobacter spp. and Pseudomonas spp. remained present until the end of the storage period. These are known spoilage organisms in fishery products. Other known spoilage organisms of crustaceans such as Photobacterium spp. could not be identified. Copyright © 2014 Elsevier Ltd. All rights reserved.

Beyond the standard plate count: genomic views into microbial food ecology

USDA-ARS?s Scientific Manuscript database

Food spoilage is a complex process that involves multiple species with specific niches and metabolic processes; bacterial culturing techniques are the traditional methods for identifying the microbes responsible. These culture-dependent methods may be considered selective, targeting the isolation of...

Microbiological hazard identification and exposure assessment of street food vending in Johannesburg, South Africa.

PubMed

Mosupye, F M; von Holy, A

2000-11-01

One hundred and thirty-two samples of beef, chicken, salad and gravy were collected from two street vendors over eleven replicate surveys to assess microbiological safety and quality. For each food type samples were collected during preparation and holding. Dish water was also collected and food preparation surfaces swabbed during preparation and display. Standard methods were used to determine aerobic plate counts, Enterobacteriaceae counts, coliform counts and spore counts. Six hundred and seventy-five predominant colonies were isolated from aerobic plate counts of all samples and characterised. The incidence of selected foodborne bacterial pathogens and non-pathogenic E. coli 1 was also determined. In most cases mean bacterial counts of the raw materials were significantly higher (P < 0.05) than those of corresponding cooked foods. No significant differences (P > 0.05) in all count types were observed between food samples collected during cooking and those collected during holding. In addition, no significant differences (P > 0.05) in all count types were observed between prepared salads and their raw materials. Mean bacterial counts of water and swab samples collected from vendor 1 were lower than those of water and swab samples collected from vendor 2.The predominant populations isolated from the aerobic plate counts were Bacillus spp., Staphylococcus spp., Enterobacteriaceae and Alcaligenes spp. Bacillus cereus was detected in 17%, Clostridium perfringens in 1%, Staphylococcus aureus in 3% and Vibrio metchnikovii in 2% of the food samples. Campylobacter jejuni, Listeria monocytogenes, and Escherichia coli O157:H7 were not detected. Non-pathogenic E. coli 1 was detected in 13% of food samples, in 86 and 36% of dish water samples collected from vendors 1 and 2, respectively, and in 36% of surface swab samples from vendor 2.

Microbial counts of mealworm larvae (Tenebrio molitor) and crickets (Acheta domesticus and Gryllodes sigillatus) from different rearing companies and different production batches.

PubMed

Vandeweyer, D; Crauwels, S; Lievens, B; Van Campenhout, L

2017-02-02

The rising interest in insects for human consumption and the changing regulations in Europe require a profound insight into the food safety of insects reared and sold in Western society. The microbial quality of edible insects has only been studied occasionally. This study aimed at generating an overview of intrinsic parameters (pH, water activity and moisture content) and microbial quality of fresh mealworm larvae and crickets for several rearing companies and for several batches per rearer. In total, 21 batches obtained from 7 rearing companies were subjected to analysis of intrinsic parameters, a range of plate counts and presence-absence tests for Salmonella spp. and Listeria monocytogenes. The microbial counts of the fresh insects were generally high. Different rearing batches from a single rearing company showed differences in microbial counts which could not be explained by variations in intrinsic properties. The largest variations were found in numbers of bacterial endospores, psychrotrophs and fungi. Salmonella spp. and L. monocytogenes were not detected in any of the samples. Altogether, our study shows that large variations were found between batches from individual rearers. As a consequence, no overall differences between rearers could be observed. Copyright © 2016 Elsevier B.V. All rights reserved.

Inactivation of Selected Bacterial Pathogens in Dairy Cattle Manure by Mesophilic Anaerobic Digestion (Balloon Type Digester)

PubMed Central

Manyi-Loh, Christy E.; Mamphweli, Sampson N.; Meyer, Edson L.; Okoh, Anthony I.; Makaka, Golden; Simon, Michael

2014-01-01

Anaerobic digestion of animal manure in biogas digesters has shown promise as a technology in reducing the microbial load to safe and recommended levels. We sought to treat dairy manure obtained from the Fort Hare Dairy Farm by investigating the survival rates of bacterial pathogens, through a total viable plate count method, before, during and after mesophilic anaerobic digestion. Different microbiological media were inoculated with different serial dilutions of manure samples that were withdrawn from the biogas digester at 3, 7 and 14 day intervals to determine the viable cells. Data obtained indicated that the pathogens of public health importance were 90%–99% reduced in the order: Campylobacter sp. (18 days) < Escherichia coli sp. (62 days) < Salmonella sp. (133 days) from a viable count of 10.1 × 103, 3.6 × 105, 7.4 × 103 to concentrations below the detection limit (DL = 102 cfu/g manure), respectively. This disparity in survival rates may be influenced by the inherent characteristics of these bacteria, available nutrients as well as the stages of the anaerobic digestion process. In addition, the highest p-value i.e., 0.957 for E. coli showed the statistical significance of its model and the strongest correlation between its reductions with days of digestion. In conclusion, the results demonstrated that the specific bacterial pathogens in manure can be considerably reduced through anaerobic digestion after 133 days. PMID:25026086

Zoonotic bacterial meningitis in human adults.

PubMed

van Samkar, Anusha; Brouwer, Matthijs C; van der Ende, Arie; van de Beek, Diederik

2016-09-13

To describe the epidemiology, etiology, clinical characteristics, treatment, outcome, and prevention of zoonotic bacterial meningitis in human adults. We identified 16 zoonotic bacteria causing meningitis in adults. Zoonotic bacterial meningitis is uncommon compared to bacterial meningitis caused by human pathogens, and the incidence has a strong regional distribution. Zoonotic bacterial meningitis is mainly associated with animal contact, consumption of animal products, and an immunocompromised state of the patient. In a high proportion of zoonotic bacterial meningitis cases, CSF analysis showed only a mildly elevated leukocyte count. The recommended antibiotic therapy differs per pathogen, and the overall mortality is low. Zoonotic bacterial meningitis is uncommon but is associated with specific complications. The suspicion should be raised in patients with bacterial meningitis who have recreational or professional contact with animals and in patients living in regions endemic for specific zoonotic pathogens. An immunocompromised state is associated with a worse prognosis. Identification of risk factors and underlying disease is necessary to improve treatment. © 2016 American Academy of Neurology.

APPARENT BIAS IN RIVER WATER INOCULUM FOLLOWING CENTRIFUGATION

EPA Science Inventory

We collected four measures of viable bacterial concentration (heterotrophic plate count, total coliform, fecal coliform, and Escherichia coli) and three measures of well color development in Biolog GN2 microtiter plates from water samples that were collected on two or three separ...

Comparison of bacterial counts obtained from naturally contaminated foods by means of Stomacher and blender.

PubMed

Purvis, U; Sharpe, A N; Bergener, D M; Lachapelle, G; Milling, M; Spiring, F

1987-01-01

Four Regional Health Protection Branch laboratories each compared aerobic colony counts obtained after "stomaching" and blending, for a minimum of 10 samples in each of the seven food groups: dry pastas; chocolate and cocoa powders; frozen entrees (macaroni and cheese, chow mein, chop suey, fried rice, seafood casseroles, and Salisbury steak); nonfat dry milk; shrimp and crabmeats; spices; and breakfast sausages. Overall, counts obtained after using the Stomacher were equivalent to or higher than counts obtained after using the blender in 73% of the comparisons (alpha = 0.05). Where differences existed, counts obtained after using the Stomacher tended to be higher than counts obtained after using the blender from milk powder and lower from sausage. Aerobic colony counts from these foods are not unacceptably biased when obtained by Stomacher.

Simplifying Collection of Corneal Specimens in Cases of Suspected Bacterial Keratitis

PubMed Central

Kaye, Stephen B.; Rao, Prasad G.; Smith, Godfrey; Scott, John A.; Hoyles, Sharon; Morton, Clare E.; Willoughby, Colin; Batterbury, Mark; Harvey, Graham

2003-01-01

Identification of the causative organisms in suspected bacterial keratitis traditionally involves collecting multiple corneal scrapes, which are plated directly onto different solid agar culture media. Difficulties have been reported with this practice, so the development of a simpler diagnostic method in suspected bacterial keratitis would be useful. It is unclear whether a single corneal scrape sent to the microbiology laboratory in a liquid transport culture medium (indirect method) is as reliable for the diagnosis of bacterial keratitis as inoculation of multiple scrapes directly onto agar plates (direct method). To investigate this, bacterial recovery was assessed following transfer and transport of different concentrations and types of bacteria from an artificially contaminated surgical blade into brain heart infusion (BHI). Bacterial recovery rates between the proposed (indirect) and standard (direct) method were then compared after the in vitro inoculation of pig corneas and following specimen collection in patients with presumed bacterial ulcerative keratitis. Recovery of bacteria from contaminated surgical blades was found to be the same from both solid and liquid culture media. There was no significant difference in the numbers of positive cultures from solid (direct) and liquid (indirect) culture media, both in the experimental pig cornea inoculation study (P = 0.34) and in experiments with patients with clinical infections (P = 0.4), with an 85.2% agreement between methods (kappa = 0.61, P < 0.0001). In conclusion, therefore, the collection of two corneal scrapes, one used for Gram staining and the other transported in BHI followed by plating and subculturing in an enrichment medium, provides a simple method for the investigation of presumed bacterial keratitis. PMID:12843063

Statistical modeling of dental unit water bacterial test kit performance.

PubMed

Cohen, Mark E; Harte, Jennifer A; Stone, Mark E; O'Connor, Karen H; Coen, Michael L; Cullum, Malford E

2007-01-01

While it is important to monitor dental water quality, it is unclear whether in-office test kits provide bacterial counts comparable to the gold standard method (R2A). Studies were conducted on specimens with known bacterial concentrations, and from dental units, to evaluate test kit accuracy across a range of bacterial types and loads. Colony forming units (CFU) were counted for samples from each source, using R2A and two types of test kits, and conformity to Poisson distribution expectations was evaluated. Poisson regression was used to test for effects of source and device, and to estimate rate ratios for kits relative to R2A. For all devices, distributions were Poisson for low CFU/mL when only beige-pigmented bacteria were considered. For higher counts, R2A remained Poisson, but kits exhibited over-dispersion. Both kits undercounted relative to R2A, but the degree of undercounting was reasonably stable. Kits did not grow pink-pigmented bacteria from dental-unit water identified as Methylobacterium rhodesianum. Only one of the test kits provided results with adequate reliability at higher bacterial concentrations. Undercount bias could be estimated for this device and used to adjust test kit results. Insensitivity to methylobacteria spp. is problematic.

Feasibility study for epidemic prevention and control in a regional hospital.

PubMed

Chen, Yung-Liang; Yeh, Ming-Yang; Huang, Shau-Yen; Liu, Chi-Ming; Sun, Chi-Chen; Lu, Hsu-Feng; Chiu, Tsan-Hung; Hsia, Te-Chun; Chung, Jing-Gung

2012-03-01

Epidemic prevention policies in hospitals address issues such as, indoor air quality control, cleanliness of medical staff clothing and employee hand-washing procedures. Our hospital employed Bio-Kil to treat air-conditioning filters and nursing staff uniforms. We also assessed the efficacy of different detergents. Using Bio-Kil technology, the mean bacterial count in the air was reduced from 108.8 CFU/h/plate (n=420) to 68.6 CFU/h/plate (n=630). On the lower hems of the Bio-Kil-treated gowns, the mean bacterial count was 1,201 CFU/100 cm(2), markedly lower than the bacterial count of 7,753 CFU/100 cm(2), found on the parts of the gowns not treated with Bio-Kil (p=0.0401). On the cuffs of sleeves treated with Bio-Kil, the mean count was 1,165 CFU/100 cm(2), markedly lower than that of 2,131 CFU/100 cm(2), found on the cuffs not treated with Bio-Kil (p=0.0073). With regard to the mean bacterial eradication rates of antimicrobial solutions, Steridal Solution, 75% alcohol and Bio-Kil (3rd generation) were shown to be the most effective, with rates exceeding 80%. Hibiscrub with paper towels and Fresh Protect Skin were the second most effective. Bio-Kil (1st generation), tap water with paper towels, liquid hand soap with paper towels and ozone water were the least effective. One important observation was that hand-washing without the use of paper towels increased the bacterial count by as much as 84% . Bio-Kil is effective in reducing bacterial counts in the air, on nursing staff uniforms and is an effective detergent.

New method for estimating bacterial cell abundances in natural samples by use of sublimation

NASA Technical Reports Server (NTRS)

Glavin, Daniel P.; Cleaves, H. James; Schubert, Michael; Aubrey, Andrew; Bada, Jeffrey L.

2004-01-01

We have developed a new method based on the sublimation of adenine from Escherichia coli to estimate bacterial cell counts in natural samples. To demonstrate this technique, several types of natural samples, including beach sand, seawater, deep-sea sediment, and two soil samples from the Atacama Desert, were heated to a temperature of 500 degrees C for several seconds under reduced pressure. The sublimate was collected on a cold finger, and the amount of adenine released from the samples was then determined by high-performance liquid chromatography with UV absorbance detection. Based on the total amount of adenine recovered from DNA and RNA in these samples, we estimated bacterial cell counts ranging from approximately 10(5) to 10(9) E. coli cell equivalents per gram. For most of these samples, the sublimation-based cell counts were in agreement with total bacterial counts obtained by traditional DAPI (4,6-diamidino-2-phenylindole) staining.

[Factors affecting the DAPI fluorescence direct count in the tidal river sediment].

PubMed

Chen, Chen; Huang, Shan; Wu, Qun-he; Li, Rui-yi; Zhang, Ren-duo

2010-08-01

The factors affecting the DAPI (4', 6-diamidino-2-phenylidole) fluorescence direct count in the tidal river sediment were examined. Sediment samples were collected from the Guangzhou section of the Pearl River. Besides sediment texture and organic matter, an improved staining procedure and the involved parameters were analyzed. Results showed that the procedure with the sediment with 2000 fold dilution and ultrasonic water bath for 10 min, and with a final DAPI concentration of 10 microg x mL(-1) and staining time for more than 30 min produced the optimum results of DAPI direct count in the sediment. The total bacterial number was correlated to the proportion of the non-nucleoid-containing cells to the total bacterial number (r = 0.587, p = 0.004). The organic matter content also correlated to the ration. The clay content had a strong correlation with the organic matter, through which the clay content also affected the ratio. A multiple regression analysis between the ration versus the organic matter, the total bacterial number, and the clay content showed that the regression equation fit the measure values satisfactorily (r = 0.694). These results indicated that the above factors needed to be considered in the applications of the DAPI fluorescence direct counting method to the tidal river sediment.

Suitability of Optical, Physical and Chemical Measurements for Detection of Changes in Bacterial Drinking Water Quality

PubMed Central

Ikonen, Jenni; Pitkänen, Tarja; Miettinen, Ilkka T.

2013-01-01

In this study, different optical, physical and chemical measurements were tested for their capacity to detect changes in water quality. The tests included UV-absorbance at 254 nm, absorbance at 420 nm, turbidity, particle counting, temperature, pH, electric conductivity (EC), free chlorine concentration and ATP concentration measurements. Special emphasis was given to investigating the potential for measurement tools to detect changes in bacterial concentrations in drinking water. Bacterial colony counts (CFU) and total bacterial cell counts (TBC) were used as reference methods for assessing the bacterial water quality. The study consists of a series of laboratory scale experiments: monitoring of regrowth of Pseudomonas fluorescens, estimation of the detection limits for optical measurements using Escherichia coli dilutions, verification of the relationships by analysing grab water samples from various distribution systems and utilisation of the measurements in the case of an accidentally contaminated distribution network. We found significant correlations between the tested measurements and the bacterial water quality. As the bacterial contamination of water often co-occurs with the intrusion of matrixes containing mainly non-bacterial components, the tested measurement tools can be considered to have the potential to rapidly detect any major changes in drinking water quality. PMID:24284353

Suitability of optical, physical and chemical measurements for detection of changes in bacterial drinking water quality.

PubMed

Ikonen, Jenni; Pitkänen, Tarja; Miettinen, Ilkka T

2013-10-25

In this study, different optical, physical and chemical measurements were tested for their capacity to detect changes in water quality. The tests included UV-absorbance at 254 nm, absorbance at 420 nm, turbidity, particle counting, temperature, pH, electric conductivity (EC), free chlorine concentration and ATP concentration measurements. Special emphasis was given to investigating the potential for measurement tools to detect changes in bacterial concentrations in drinking water. Bacterial colony counts (CFU) and total bacterial cell counts (TBC) were used as reference methods for assessing the bacterial water quality. The study consists of a series of laboratory scale experiments: monitoring of regrowth of Pseudomonas fluorescens, estimation of the detection limits for optical measurements using Escherichia coli dilutions, verification of the relationships by analysing grab water samples from various distribution systems and utilisation of the measurements in the case of an accidentally contaminated distribution network. We found significant correlations between the tested measurements and the bacterial water quality. As the bacterial contamination of water often co-occurs with the intrusion of matrixes containing mainly non-bacterial components, the tested measurement tools can be considered to have the potential to rapidly detect any major changes in drinking water quality.

The development and evaluation of ultrasound for the treatment of bacterial suspensions. A study of frequency, power and sonication time on cultured Bacillus species.

PubMed

Joyce, E; Phull, S S; Lorimer, J P; Mason, T J

2003-10-01

Some species of bacteria produce colonies and spores which agglomerate in spherical clusters (Bacillus subtilis) and this serves as a protection for the organisms inside against biocidal attack. Flocs of fine particles e.g. clay can entrap bacteria which can also protect them against the biocides. It is because of problems such as these that alternative methods of disinfecting water are under active investigation. One such method is the use of power ultrasound, either alone or in combination with other methods. Ultrasound is able to inactivate bacteria and deagglomerate bacterial clusters or flocs through a number of physical, mechanical and chemical effects arising from acoustic cavitation. The aim of this study was to investigate the effect of power ultrasound at different powers and frequencies on Bacillus subtilis. Viable plate count techniques were used as a measure of microbial activity. Results showed a significant increase in percent kill for Bacillus species with increasing duration of exposure and intensity of ultrasound in the low-kilohertz range (20 and 38 kHz). Results obtained at two higher frequencies (512 and 850 kHz) indicated a significant increase in bacteria count suggesting declumping. In assessing the bacterial kill with time under different sonication regimes three types of behaviour were characterized: High power ultrasound (lower frequencies) in low volumes of bacterial suspension results in a continuous reduction in bacterial cell numbers i.e. the kill rate predominates. High power ultrasound (lower frequencies) in larger volumes results in an initial rise in cell numbers suggesting declumping of the bacteria but this initial rise then falls as the declumping finishes and the kill rate becomes more important. Low intensity ultrasound (higher frequencies) gives an initial rise in cell numbers as a result of declumping. The kill rate is low and so there is no significant subsequent decrease in bacterial cell numbers.

Bacterial communities in the collection and chlorinated distribution sections of a drinking water system in Budapest, Hungary.

PubMed

Homonnay, Zalán G; Török, György; Makk, Judit; Brumbauer, Anikó; Major, Eva; Márialigeti, Károly; Tóth, Erika

2014-07-01

Bacterial communities of a bank-filtered drinking water system were investigated by aerobic cultivation and clone library analysis. Moreover, bacterial communities were compared using sequence-aided terminal restriction fragment length polymorphism (T-RFLP) fingerprinting at ten characteristic points located at both the collecting and the distributing part of the water supply system. Chemical characteristics of the samples were similar, except for the presence of chlorine residuals in the distribution system and increased total iron concentration in two of the samples. Assimilable organic carbon (AOC) concentration increased within the collection system, it was reduced by chlorination and it increased again in the distribution system. Neither fecal indicators nor pathogens were detected by standard cultivation techniques. Chlorination reduced bacterial diversity and heterotrophic plate counts. Community structures were found to be significantly different before and after chlorination: the diverse communities in wells and the collection system were dominated by chemolithotrophic (e.g., Gallionella and Nitrospira) and oligocarbophilic-heterotrophic bacteria (e.g., Sphingomonas, Sphingopyxis, and Bradyrhizobium). After chlorination in the distribution system, the most characteristic bacterium was related to the facultative methylotrophic Methylocella spp. Communities changed within the distribution system too, Mycobacterium spp. or Sphingopyxis spp. became predominant in certain samples. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A meta-barcoding approach to assess and compare the storage temperature-dependent bacterial diversity of gilt-head sea bream (Sparus aurata) originating from fish farms from two geographically distinct areas of Greece.

PubMed

Parlapani, Foteini F; Michailidou, Sofia; Pasentsis, Konstantinos; Argiriou, Anagnostis; Krey, Grigorios; Boziaris, Ioannis S

2018-08-02

Bacterial diversity of whole gilt-head sea bream (Sparus aurata L. 1758) originating from Ionian and Aegean Sea aquaculture farms and stored at 0 (ice), 4 and 8 °C was determined by 16S rRNA gene amplicon sequencing method using the Illumina's MiSeq platform. The composition of Aerobic Plate Counts (APC) was also monitored by 16S rRNA gene sequencing. The rejection time point of sea bream from either area, as determined by sensory evaluation, was about 14, 6 and 3 days at 0, 4 and 8 °C, respectively. APC was approximately 4.5 log cfu/g at day 0 and ranged from 7.5 to 8.5 log cfu/g at sensory rejection. Culture-depended analysis showed that Pseudomonas and Shewanella were the most abundant microorganisms grown on plates for both seas. Moreover, culture-independent analysis of DNA extracted directly from fish flesh showed that sea bream originating from different geographical areas exhibited different bacterial diversity. Pseudomonas and Psychrobacter were the dominant microorganisms of chill-stored fish from Ionian (apart from 8 °C, where Carnobacterium dominated) and Aegean Sea, respectively. In addition, small changes of storage temperature greatly affected bacterial microbiota of stored fish. Various bacterial species, not detected by conventional microbiological methods, were also revealed through 16S amplicon sequencing. In conclusion, the use of NGS approach is a promising methodology for assessing bacterial diversity of sea bream originating from different geographical areas and stored at various temperatures. Copyright © 2018 Elsevier B.V. All rights reserved.

Establishment of HPC(R2A) for regrowth control in non-chlorinated distribution systems.

PubMed

Uhl, Wolfgang; Schaule, Gabriela

2004-05-01

Drinking water distributed without disinfection and without regrowth problems for many years may show bacterial regrowth when the residence time and/or temperature in the distribution system increases or when substrate and/or bacterial concentration in the treated water increases. An example of a regrowth event in a major German city is discussed. Regrowth of HPC bacteria occurred unexpectedly at the end of a very hot summer. No pathogenic or potentially pathogenic bacteria were identified. Increased residence times in the distribution system and temperatures up to 25 degrees C were identified as most probable causes and the regrowth event was successfully overcome by changing flow regimes and decreasing residence times. Standard plate counts of HPC bacteria using the spread plate technique on nutrient rich agar according to German Drinking Water Regulations (GDWR) had proven to be a very good indicator of hygienically safe drinking water and to demonstrate the effectiveness of water treatment. However, the method proved insensitive for early regrowth detection. Regrowth experiments in the lab and sampling of the distribution system during two summers showed that spread plate counts on nutrient-poor R2A agar after 7-day incubation yielded 100 to 200 times higher counts. Counts on R2A after 3-day incubation were three times less than after 7 days. As the precision of plate count methods is very poor for counts less than 10 cfu/plate, a method yielding higher counts is better suited to detect upcoming regrowth than a method yielding low counts. It is shown that for the identification of regrowth events HPC(R2A) gives a further margin of about 2 weeks for reaction before HPC(GDWR). Copyright 2003 Elsevier B.V.

Comparison of bacterial counts in expressed breast milk following standard or strict infection control regimens in neonatal intensive care units: compliance of mothers does matter.

PubMed

Haiden, N; Pimpel, B; Assadian, O; Binder, C; Kreissl, A; Repa, A; Thanhäuser, M; Roberts, C D; Berger, A

2016-03-01

Bacterial counts in 1466 expressed breast milk (EBM) samples from women following one of two infection control regimens (standard vs strict) were investigated. Overall, 12% of samples yielded Gram-negative bacteria, with no significant differences between the standard [11.9% (94/788)] and strict [12.1% (82/678)] regimens (P = 0.92). Significantly more samples were contaminated when expressed at home (standard regimen home/hospital: 17.9% vs 6.1%; strict regimen home/hospital: 19.6% vs 3.4%; P < 0.001). Bacterial contamination of EBM was not associated with the regimen, but was associated with the location of breast milk expression. Attempts to improve personal hygiene during milk collection seem to be of limited value. Good hygiene of collection and storage equipment is likely to be the most important way to ensure the microbiological quality of EBM. Copyright © 2016 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

Defective plastic infection-control barriers and faulty technique may cause PSP plate contamination used in digital intraoral radiography.

PubMed

Kuperstein, Arthur S

2012-09-01

Fifty-two disinfected photostimulable phosphor (PSP) plates in plastic barrier envelopes were evaluated for contamination following placement in 30 study participants. Forty-four plates were acceptable for use in the study. The risk factor was the abundant oropharyngeal microbial flora and its ability to breach infection-control barrier sheaths. The presence of bacterial colonies on an agar plate was used to determine bacterial contamination and the presence of any growth indicated failure of the barrier envelope. Before clinical placement of the plates, quality review of the PSP plates revealed defects in the integrity of 4 barrier envelopes most likely caused by forceps-related damage or failure to achieve a uniform seal during manufacturing. These defects allowed substantial contamination. Contamination also occurred as a result of failure to extract the PSP plate from the barrier envelope cleanly. Of the 44 barriers with no obvious signs of a defect, 3 produced bacterial growth following culture. The authors concluded that digital sensor sheathed in barrier envelopes remain a potential source of contamination. PSP plates must be disinfected between removal from a contaminated barrier envelope (used in a patient) and placement in a new barrier envelope. In addition, placement into the barrier envelope should ideally be carried out under aseptic conditions. Finally, the integrity of each sealed barrier envelope must be verified visually. Copyright © 2012. Published by Mosby, Inc. All rights reserved.

Extinction map of Chamaeleon I molecular cloud with DENIS star counts.

NASA Astrophysics Data System (ADS)

Cambresy, L.; Epchtein, N.; Copet, E.; de Batz, B.; Kimeswenger, S.; Le Bertre, T.; Rouan, D.; Tiphene, D.

1997-08-01

Massive, large scale star counts in the J (1.25μm) band provided by the Deep Near Infrared Survey of the Southern Sky (DENIS) are used for the first time to draw out an extinction map of the Chamaeleon I dark cloud. We derived a 2' resolution map of the cloud from J star counts within an area of 1.5°x3° around the centre of the cloud using an adaptive grid method and applying a wavelet decomposition. Possible contaminating young stellar objects within the cloud are removed, although they are shown to have a negligible effect on the counts. A comparison of our extinction map with the cold contribution of the IRAS 100μm emission shows an almost perfect matching. It is shown that J star counts supersede optical counts on Schmidt plate where A_V_>4.

Soybean extracts facilitate bacterial agglutination and prevent biofilm formation on orthodontic wire.

PubMed

Lee, Heon-Jin; Kwon, Tae-Yub; Kim, Kyo-Han; Hong, Su-Hyung

2014-01-01

Soybean is an essential food ingredient that contains a class of organic compounds known as isoflavones. It is also well known that several plant agglutinins interfere with bacterial adherence to smooth surfaces. However, little is known about the effects of soybean extracts or genistein (a purified isoflavone from soybean) on bacterial biofilm formation. We evaluated the effects of soybean (Glycine max) extracts, including fermented soybean and genistein, on streptococcal agglutination and attachment onto stainless steel orthodontic wire. After cultivating streptococci in biofilm medium containing soybean extracts and orthodontic wire, the viable bacteria attached to the wire were counted. Phase-contrast microscopy and scanning electron microscopy (SEM) analyses were conducted to evaluate bacterial agglutination and attachment. Our study showed that soybean extracts induce agglutination between streptococci, which results in bacterial precipitation. Conversely, viable bacterial counting and SEM image analysis of Streptococcus mutans attached to the orthodontic wire show that bacterial attachment decreases significantly when soybean extracts were added. However, there was no significant change in pre-attached S. mutans biofilm in response to soybean. A possible explanation for these results is that increased agglutination of planktonic streptococci by soybean extracts results in inhibition of bacterial attachment onto the orthodontic wire.

Moxifloxacin superior to cefuroxime in reducing bacterial adhesion of Staphylococcus epidermidis on hydrophobic intraocular lenses.

PubMed

Benbouzid, Fathalah; Kodjikian, Laurent; Hartmann, Daniel; Renaud, François; Baillif, Stéphanie

2016-02-01

To compare the anti-adhesive effect of cefuroxime and moxifloxacin on the primary attachment phase of Staphylococcus epidermidis on hydrophobic acrylic intraocular lenses (IOLs). Forty hydrophobic acrylic IOLs were used. Two groups of IOLs were soaked in a moxifloxacin (Mox-T1: 0.5 mg/0.1 ml) or a cefuroxime (Cef-T1: cefuroxime 1 mg/0.1 ml) solution before incubation in a S. epidermidis bacterial suspension. Two other groups were incubated in the bacterial suspension before antibiotics (Cef-T2 and Mox-T2) were added. The control group (Ctrl) consisted of IOLs incubated in the bacterial suspension. After incubation, IOLs were sonicated and vortexed. The resultant suspension was spread over a nutritive agar plate. Bacterial colonies were counted after 24 hr of incubation. Mean number of colony-forming units per IOL was Cef-T1: 184 × 10(3) (SE: 5.24; SD: 28.21), Cef-T2: 117 × 10(3) (SE: 5.74; SD: 30.37), Mox-T1: 1.27 × 10(3) (SE: 0.12; SD: 0.61), Mox-T2: 25 × 10(3) (SE:1.98; SD: 9.72) and Ctrl: 361 × 10(3) (SE: 26.9; SD: 107.6). The number of adhering bacteria did not vary whether cefuroxime was added before or after IOL incubation in the bacterial suspension (p = 0.132). Moxifloxacin was more effective in reducing the number of adhering bacteria when used before IOL incubation (p < 0.001). Overall for T1 and T2, moxifloxacin was more effective than cefuroxime in reducing bacterial adhesion on IOLs (p < 0.001). Moxifloxacin and cefuroxime significantly reduced S. epidermidis adhesion on hydrophobic acrylic IOLs. The anti-adhesive effect was superior with moxifloxacin. © 2015 Acta Ophthalmologica Scandinavica Foundation. Published by John Wiley & Sons Ltd.

Reduction of pasteurization temperature leads to lower bacterial outgrowth in pasteurized fluid milk during refrigerated storage: a case study.

PubMed

Martin, N H; Ranieri, M L; Wiedmann, M; Boor, K J

2012-01-01

Bacterial numbers over refrigerated shelf-life were enumerated in high-temperature, short-time (HTST) commercially pasteurized fluid milk for 15 mo before and 15 mo after reducing pasteurization temperature from 79.4°C (175°F) [corrected] to 76.1°C (169°F). Total bacterial counts were measured in whole fat, 2% fat, and fat-free milk products on the day of processing as well as throughout refrigerated storage (6°C) at 7, 14, and 21 d postprocessing. Mean total bacterial counts were significantly lower immediately after processing as well as at 21 d postprocessing in samples pasteurized at 76.1°C versus samples pasteurized at 79.4°C. In addition to mean total bacterial counts, changes in bacterial numbers over time (i.e., bacterial growth) were analyzed and were lower during refrigerated storage of products pasteurized at the lower temperature. Lowering the pasteurization temperature for unflavored fluid milk processed in a commercial processing facility significantly reduced bacterial growth during refrigerated storage. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Low noise and conductively cooled microchannel plates

NASA Technical Reports Server (NTRS)

Feller, W. B.

1990-01-01

Microchannel plate (MCP) dynamic range has recently been enhanced for both very low and very high input flux conditions. Improvements in MCP manufacturing technology reported earlier have led to MCPs with substantially reduced radioisotope levels, giving dramatically lower internal background-counting rates. An update is given on the Galileo low noise MCP. Also, new results in increasing the MCP linear counting range for high input flux densities are presented. By bonding the active face of a very low resistance MCP (less than 1 megaohm) to a substrate providing a conductive path for heat transport, the bias current limit (hence, MCP output count rate limit) can be increased up to two orders of magnitude. Normal pulse-counting MCP operation was observed at bias currents of several mA when a curved-channel MCP (80:1) was bonded to a ceramic multianode substrate; the MCP temperature rise above ambient was less than 40 C.

An evaluation of bacterial contamination of barriers used in periapical tissue regeneration: Part 1--Bacterial adherence.

PubMed

Sharma, Priya; Mickel, André K; Chogle, Sami; Sharma, Prem Nath; Han, Yiping W; Jones, Jefferson J

2008-02-01

To compare the adherence of Prevotella melaninogenica and Enterococcus faecalis to 3 guided tissue regeneration membranes: Atrisorb, Lambone, and OsseoQuest. It was hypothesized that OsseoQuest would show increased bacterial adherence compared to Lambone and Atrisorb. The barriers were suspended in trypticase soy broth containing an inoculum of either P melaninogenica or E faecalis. The samples were incubated under appropriate conditions for 6, 24, and 48 hours. Following incubation, each membrane was mixed in fresh media in a vortex machine to dislodge adherent bacteria. The vortexed media was quantitatively assessed using serial dilutions for viable cell count. E faecalis exhibited higher adherence compared to P melaninogenica with time. Of the membranes tested, Lambone displayed the least bacterial adherence. An analysis of the results indicated that bacterial adherence was time-dependent for all membranes. Membrane structure, chemical configuration, hydrophobicity, and bacterial cell surface structure were suggested as factors contributing to variance in bacterial adherence.

High precision refractometry based on Fresnel diffraction from phase plates.

PubMed

Tavassoly, M Taghi; Naraghi, Roxana Rezvani; Nahal, Arashmid; Hassani, Khosrow

2012-05-01

When a transparent plane-parallel plate is illuminated at a boundary region by a monochromatic parallel beam of light, Fresnel diffraction occurs because of the abrupt change in phase imposed by the finite change in refractive index at the plate boundary. The visibility of the diffraction fringes varies periodically with changes in incident angle. The visibility period depends on the plate thickness and the refractive indices of the plate and the surrounding medium. Plotting the phase change versus incident angle or counting the visibility repetition in an incident-angle interval provides, for a given plate thickness, the refractive index of the plate very accurately. It is shown here that the refractive index of a plate can be determined without knowing the plate thickness. Therefore, the technique can be utilized for measuring plate thickness with high precision. In addition, by installing a plate with known refractive index in a rectangular cell filled with a liquid and following the described procedures, the refractive index of the liquid is obtained. The technique is applied to measure the refractive indices of a glass slide, distilled water, and ethanol. The potential and merits of the technique are also discussed.

A New Method for Estimating Bacterial Abundances in Natural Samples using Sublimation

NASA Technical Reports Server (NTRS)

Glavin, Daniel P.; Cleaves, H. James; Schubert, Michael; Aubrey, Andrew; Bada, Jeffrey L.

2004-01-01

We have developed a new method based on the sublimation of adenine from Escherichia coli to estimate bacterial cell counts in natural samples. To demonstrate this technique, several types of natural samples including beach sand, seawater, deep-sea sediment, and two soil samples from the Atacama Desert were heated to a temperature of 500 C for several seconds under reduced pressure. The sublimate was collected on a cold finger and the amount of adenine released from the samples then determined by high performance liquid chromatography (HPLC) with UV absorbance detection. Based on the total amount of adenine recovered from DNA and RNA in these samples, we estimated bacterial cell counts ranging from approx. l0(exp 5) to l0(exp 9) E. coli cell equivalents per gram. For most of these samples, the sublimation based cell counts were in agreement with total bacterial counts obtained by traditional DAPI staining. The simplicity and robustness of the sublimation technique compared to the DAPI staining method makes this approach particularly attractive for use by spacecraft instrumentation. NASA is currently planning to send a lander to Mars in 2009 in order to assess whether or not organic compounds, especially those that might be associated with life, are present in Martian surface samples. Based on our analyses of the Atacama Desert soil samples, several million bacterial cells per gam of Martian soil should be detectable using this sublimation technique.

Cerebrospinal fluid cytokines in the diagnosis of bacterial meningitis in infants.

PubMed

Srinivasan, Lakshmi; Kilpatrick, Laurie; Shah, Samir S; Abbasi, Soraya; Harris, Mary C

2016-10-01

Bacterial meningitis poses diagnostic challenges in infants. Antibiotic pretreatment and low bacterial density diminish cerebrospinal fluid (CSF) culture yield, while laboratory parameters do not reliably identify bacterial meningitis. Pro and anti-inflammatory cytokines are elevated in bacterial meningitis and may be useful diagnostic adjuncts when CSF cultures are negative. In a prospective cohort study of infants, we used cytometric bead arrays to measure tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1), IL-6, IL-8, IL-10, and IL-12 in CSF. Receiver operating characteristic (ROC) analyses and Principal component analysis (PCA) were used to determine cytokine combinations that identified bacterial meningitis. Six hundred and eighty four infants < 6 mo were included; 11 had culture-proven bacterial meningitis. IL-6 and IL-10 were the individual cytokines possessing greatest accuracy in diagnosis of culture proven bacterial meningitis (ROC analyses; area under the concentration-time curve (AUC) 0.91; 0.9103 respectively), and performed as well as, or better than combinations identified using ROC and PCA. CSF cytokines were highly correlated with each other and with CSF white blood cell count (WBC) counts in infants with meningitis. A subset of antibiotic pretreated culture-negative subjects demonstrated cytokine patterns similar to culture positive subjects. CSF cytokine levels may aid diagnosis of bacterial meningitis, and facilitate decision-making regarding treatment for culture negative meningitis.

[Spontaneous bacterial peritonitis].

PubMed

Velkey, Bálint; Vitális, Eszter; Vitális, Zsuzsanna

2017-01-01

Spontaneous bacterial peritonitis occurs most commonly in cirrhotic patients with ascites. Pathogens get into the circulation by intestinal translocation and colonize in peritoneal fluid. Diagnosis of spontaneous bacterial peritonitis is based on elevated polymorphonuclear leukocyte count in the ascites (>0,25 G/L). Ascites culture is often negative but aids to get information about antibiotic sensitivity in positive cases. Treatment in stable patient can be intravenous then orally administrated ciprofloxacin or amoxicillin/clavulanic acid, while in severe cases intravenous III. generation cephalosporin. Nosocomial spontaneous bacterial peritonitis often caused by Gram-positive bacteria and multi-resistant pathogens can also be expected thus carbapenem should be the choice of the empiric treatment. Antibiotic prophylaxis should be considered. Norfloxacin is used most commonly, but changes are expected due to increase in quinolone resistance. As a primary prophylaxis, a short-term antibiotic treatment is recommended after gastrointestinal bleeding for 5 days, while long-term prophylaxis is for patients with low ascites protein, and advanced disease (400 mg/day). Secondary prophylaxis is recommended for all patients recovered from spontaneous bacterial peritonitis. Due to increasing antibiotic use of antibiotics prophylaxis is debated to some degree. Orv. Hetil., 2017, 158(2), 50-57.

Effects of open drainage ditch design on bacterial and fungal communities of cold waterlogged paddy soils

PubMed Central

Qiu, Shanlian; Wang, MK; Wang, Fei; Chen, Jichen; Li, Xiaoyan; Li, Qinghua; Lin, Cheng; Lin, Xinjian

2013-01-01

A field experiment established in 1980 was conducted to evaluate the effects of open drainage ditch applied for water removal on bacterial and fungal communities of cold waterlogged paddy soils in 2011. In this experiment, traditional plate counting and temperature gradient gel electrophoresis were employed to characterize the abundance and diversity of soil bacterial and fungal communities. Four different distances from the open drainage ditch, 5, 15, 25 and 75 m with different degrees of drainage were designed for this study. Maximum populations of culturable aerobic bacteria and fungi were at 15-m distance while minimum populations were at 75-m distance. Significant differences (p < 0.05) in fungal populations were observed at all distances from open drainage ditch. The highest diversity of the bacterial community was found at a distance of 25 m, while that of the fungal community was observed at a distance of 5 m. Sequencing of excised TGGE bands indicated that the dominant bacteria at 75-m distance belonged to anaerobic or microaerobic bacteria. Relationships between microbial characteristics and soil physicochemical properties indicated that soil pH and available nitrogen contents were key factors controlling the abundance of culturable aerobic bacteria and fungi, while soil water capacity also affected the diversity of fungal community. These findings can provide the references for better design and advanced management of the drainage ditches in cold waterlogged paddy soils. PMID:24516468

Track counts as indices to abundances of arboreal rodents.

Treesearch

A.B. Carey; J.W. Witt

1991-01-01

Counting tracks to obtain an index of abundance for species difficult to capture offers a promise of efficiency and effectiveness when broad surveys of populations are necessary. Sand plots, smoked kymograph paper, and, recently, smoked aluminum plates have been used to record tracks(Raphael et al., 1986; Taylor and Raphael, 1988). Findings of studies of carnivores...

Bacterial and chemical oxidation of pyritic mine tailings at low temperatures

NASA Astrophysics Data System (ADS)

Elberling, Bo; Schippers, Axel; Sand, Wolfgang

2000-02-01

Microbial and chemical sulfide oxidation activity and oxygen consumption was investigated in the active layer of pyritic mine tailings at Nanisivik Mine, located in a permafrost area on Baffin Island in northern Canada. Samples of tailings were collected up to a depth of 60 cm in mid-August 1998 at 4 sites, for which the metabolic activity of sulfur- and iron-oxidizing leaching bacteria besides the chemical pyrite oxidation activity were measured on 39 tailings samples and 7 samples from a natural pyritic site by calorimetry. The tailings of varying age and water content were deposited under alkaline conditions. In situ oxygen uptake rates were measured at the tailings surface every third day, prior to sampling. In addition, cell counts of iron(II), sulfur, and thiosulfate oxidizing, lithotrophic bacteria and chemoorganotrophic microorganisms were determined quantitatively by the most-probable-number technique or by agar-plating. Results show consistent pyrite oxidation rates based on in situ oxygen uptake rates, and laboratory heat output measurements. Litho- and organotrophic bacteria were found in the tailings. Calorimetric measurements revealed that the present bacterial activity is responsible for approximately one third of the ongoing oxidation. Although leaching bacteria have previously been found in the Arctic, this study is the first to prove the significance of bacterial activity in the overall pollution resulting from tailings deposited in the Arctic.

Identification of Bacterial Species in Kuwaiti Waters Through DNA Sequencing

NASA Astrophysics Data System (ADS)

Chen, K.

2017-01-01

With an objective of identifying the bacterial diversity associated with ecosystem of various Kuwaiti Seas, bacteria were cultured and isolated from 3 water samples. Due to the difficulties for cultured and isolated fecal coliforms on the selective agar plates, bacterial isolates from marine agar plates were selected for molecular identification. 16S rRNA genes were successfully amplified from the genome of the selected isolates using Universal Eubacterial 16S rRNA primers. The resulted amplification products were subjected to automated DNA sequencing. Partial 16S rDNA sequences obtained were compared directly with sequences in the NCBI database using BLAST as well as with the sequences available with Ribosomal Database Project (RDP).

Characterization of reticulated vitreous carbon foam using a frisch-grid parallel-plate ionization chamber

NASA Astrophysics Data System (ADS)

Edwards, Nathaniel S.; Conley, Jerrod C.; Reichenberger, Michael A.; Nelson, Kyle A.; Tiner, Christopher N.; Hinson, Niklas J.; Ugorowski, Philip B.; Fronk, Ryan G.; McGregor, Douglas S.

2018-06-01

The propagation of electrons through several linear pore densities of reticulated vitreous carbon (RVC) foam was studied using a Frisch-grid parallel-plate ionization chamber pressurized to 1 psig of P-10 proportional gas. The operating voltages of the electrodes contained within the Frisch-grid parallel-plate ionization chamber were defined by measuring counting curves using a collimated 241Am alpha-particle source with and without a Frisch grid. RVC foam samples with linear pore densities of 5, 10, 20, 30, 45, 80, and 100 pores per linear inch were separately positioned between the cathode and anode. Pulse-height spectra and count rates from a collimated 241Am alpha-particle source positioned between the cathode and each RVC foam sample were measured and compared to a measurement without an RVC foam sample. The Frisch grid was positioned in between the RVC foam sample and the anode. The measured pulse-height spectra were indiscernible from background and resulted in negligible net count rates for all RVC foam samples. The Frisch grid parallel-plate ionization chamber measurement results indicate that electrons do not traverse the bulk of RVC foam and consequently do not produce a pulse.

Evaluation of the Dark-Medium Objective Lens in Counting Asbestos Fibers by Phase-Contrast Microscopy

PubMed Central

Lee, Eun Gyung; Nelson, John H.; Kashon, Michael L.; Harper, Martin

2015-01-01

A Japanese round-robin study revealed that analysts who used a dark-medium (DM) objective lens reported higher fiber counts from American Industrial Hygiene Association (AIHA) Proficiency Analytical Testing (PAT) chrysotile samples than those using a standard objective lens, but the cause of this difference was not investigated at that time. The purpose of this study is to determine any major source of this difference by performing two sets of round-robin studies. For the first round-robin study, 15 AIHA PAT samples (five each of chrysotile and amosite generated by water-suspended method, and five chrysotile generated by aerosolization method) were prepared with relocatable cover slips and examined by nine laboratories. A second round-robin study was then performed with six chrysotile field sample slides by six out of nine laboratories who participated in the first round-robin study. In addition, two phase-shift test slides to check analysts’ visibility and an eight-form diatom test plate to compare resolution between the two objectives were examined. For the AIHA PAT chrysotile reference slides, use of the DM objective resulted in consistently higher fiber counts (1.45 times for all data) than the standard objective (P-value < 0.05), regardless of the filter generation (water-suspension or aerosol) method. For the AIHA PAT amosite reference and chrysotile field sample slides, the fiber counts between the two objectives were not significantly different. No statistically significant differences were observed in the visibility of blocks of the test slides between the two objectives. Also, the DM and standard objectives showed no pattern of differences in viewing the fine lines and/or dots of each species images on the eight-form diatom test plate. Among various potential factors that might affect the analysts’ performance of fiber counts, this study supports the greater contrast caused by the different phase plate absorptions as the main cause of high counts for

Bacterial aerosol emission rates from municipal wastewater aeration tanks.

PubMed Central

Sawyer, B; Elenbogen, G; Rao, K C; O'Brien, P; Zenz, D R; Lue-Hing, C

1993-01-01

In this report we describe the results of a study conducted to determine the rates of bacterial aerosol emission from the surfaces of the aeration tanks of the Metropolitan Water Reclamation District of Greater Chicago John E. Egan Water Reclamation Plant. This study was accomplished by conducting test runs in which Andersen six-stage viable samplers were used to collect bacterial aerosol samples inside a walled tower positioned above an aeration tank liquid surface at the John E. Egan Water Reclamation Plant. The samples were analyzed for standard plate counts (SPC), total coliforms (TC), fecal coliforms, and fecal streptococci. Two methods of calculation were used to estimate the bacterial emission rate. The first method was a conventional stack emission rate calculation method in which the measured air concentration of bacteria was multiplied by the air flow rate emanating from the aeration tanks. The second method was a more empirical method in which an attempt was made to measure all of the bacteria emanating from an isolated area (0.37 m2) of the aeration tank surface over time. The data from six test runs were used to determine bacterial emission rates by both calculation methods. As determined by the conventional calculation method, the average SPC emission rate was 1.61 SPC/m2/s (range, 0.66 to 2.65 SPC/m2/s). As determined by the empirical calculation method, the average SPC emission rate was 2.18 SPC/m2/s (range, 1.25 to 2.66 SPC/m2/s). For TC, the average emission rate was 0.20 TC/m2/s (range, 0.02 to 0.40 TC/m2/s) when the conventional calculation method was used and 0.27 TC/m2/s (range, 0.04 to 0.53 TC/m2/s) when the empirical calculation method was used.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8250547

Evolution of bacterial communities in the Gironde Estuary (France) according to a salinity gradient

NASA Astrophysics Data System (ADS)

Prieur, D.; Troussellier, M.; Romana, A.; Chamroux, S.; Mevel, G.; Baleux, B.

1987-01-01

Three surveys were performed in the Gironde Estuary (France) in August 1981, March 1982 and July 1982. For each campaign, seventy samples were taken by helicopter, in order to follow the tide along the estuary. Of the parameters that were studied, salinity appeared to be the most important and which controls the bacterial communities along the estuary. This paper deals with the evolution of bacterial communities along a salinity gradient. The information obtained from various bacteriological parameters (total bacterial counts, viable counts on salted and unsalted media, functional evenness) were convergent. The bacterial community is dominated by an halotolerant microflora. In the estuary, a continental microflora is followed by a marine microflora. The succession zone between these two microflora is located between 5 and 10‰ areas of salinity.

Bacterial flora of the sigmoid neovagina.

PubMed Central

Toolenaar, T A; Freundt, I; Wagenvoort, J H; Huikeshoven, F J; Vogel, M; Jeekel, H; Drogendijk, A C

1993-01-01

The bacterial microbiota of 15 sigmoid neovaginas, created in patients with congenital vaginal aplasia or male transsexualism, was studied. No specimen was sterile, and only normal inhabitants of the colon were cultured. The total counts of bacteria were lower than those reported for healthy sigmoid colons. PMID:8308126

Photon-counting image sensors for the ultraviolet

NASA Technical Reports Server (NTRS)

Jenkins, E. B.

1985-01-01

An investigation on specific performance details of photon counting, ultraviolet image sensors having 2-dimensional formats is reviewed. In one study, controlled experiments were performed which compare the quantum efficiencies, in pulse counting mode, of CsI photocathodes deposited on: (1) the front surface of a microchannel plate (MCP), (2) a solid surface in front of an MCP, and (3) an intensified CCD image sensor (ICCD) where a CCD is directly bombarded by accelerated photoelectrons. Tests indicated that the detection efficiency of the CsI-coated MCP at 1026 A is lower by a factor of 2.5 than that of the MCP with a separate, opaque CsI photocathode, and the detection efficiency ratio increases substantially at longer wavelengths (ratio is 5 at 1216 A and 20 at 1608 A).

Evidence of variable bacterial colonization on coloured elastomeric ligatures during orthodontic treatment: An intermodular comparative study.

PubMed

Sharma, Ravish; Sharma, Kavita; Sawhney, Rajesh

2018-03-01

Besides, other factors, the choice of materials used as orthodontic ligatures could be one of the many tools to counter the effects of microbial adhesion, that culminates into dental ailments. Therefore, we assessed bacterial adhesion on elastomeric ligatures with special reference to coloured elastomeric rings during orthodontic treatment. A split mouth study, involving 240 samples of different elastomeric ligatures from forty orthodontic patients possessing good oral hygiene was carried out. The archwire was ligated to the brackets on both arches with elastomeric rings (superslick, clear transparent , blue and pink) at predetermined quadrants. After six weeks, ligatures from second premolars were removed and processed for bacterial enumeration using standard techniques. Bacterial counts were also determined for stimulated saliva samples taken at 0 and 6 weeks. A statistically significant difference in bacterial counts was obtained amongst different elastomeric modules used. Maximum bacterial counts were found on conventional pigmented elastomeric modules, followed by Superslick module and clear module. More number of bacteria associated with the conventional pink as compared to the conventional blue pigmented modules, however it was not statistically significant. The three bacterial genera Streptococcus Staphylococcus and Aerobic Lactobacilli adhered to elastomeric modules in following predominant pattern i.e. Conventional pink>Conventional Blue>Superslick>Clear. The studies evidenced colour and material dependent bacterial colonization on orthodontic modules and could be an indicator of bacterial biofilm forming potential based on surface chemistries and a clinically efficacious tool to redesign conventional and modified elastomeric rings as orthodontic ligation accessories. Key words: Bacterial colonization, biofilm, coloured elastomers, orthodontic ligatures.

Reflected scatterometry for noninvasive interrogation of bacterial colonies

USDA-ARS?s Scientific Manuscript database

A phenotyping of bacterial colonies on agar plates using forward-scattering diffraction-pattern analysis provided promising classification of several different bacteria such as Salmonella, Vibrio, Listeria, and E. coli. Since the technique is based on forward-scattering phenomena, light transmittanc...

SPARTA: Simple Program for Automated reference-based bacterial RNA-seq Transcriptome Analysis.

PubMed

Johnson, Benjamin K; Scholz, Matthew B; Teal, Tracy K; Abramovitch, Robert B

2016-02-04

Many tools exist in the analysis of bacterial RNA sequencing (RNA-seq) transcriptional profiling experiments to identify differentially expressed genes between experimental conditions. Generally, the workflow includes quality control of reads, mapping to a reference, counting transcript abundance, and statistical tests for differentially expressed genes. In spite of the numerous tools developed for each component of an RNA-seq analysis workflow, easy-to-use bacterially oriented workflow applications to combine multiple tools and automate the process are lacking. With many tools to choose from for each step, the task of identifying a specific tool, adapting the input/output options to the specific use-case, and integrating the tools into a coherent analysis pipeline is not a trivial endeavor, particularly for microbiologists with limited bioinformatics experience. To make bacterial RNA-seq data analysis more accessible, we developed a Simple Program for Automated reference-based bacterial RNA-seq Transcriptome Analysis (SPARTA). SPARTA is a reference-based bacterial RNA-seq analysis workflow application for single-end Illumina reads. SPARTA is turnkey software that simplifies the process of analyzing RNA-seq data sets, making bacterial RNA-seq analysis a routine process that can be undertaken on a personal computer or in the classroom. The easy-to-install, complete workflow processes whole transcriptome shotgun sequencing data files by trimming reads and removing adapters, mapping reads to a reference, counting gene features, calculating differential gene expression, and, importantly, checking for potential batch effects within the data set. SPARTA outputs quality analysis reports, gene feature counts and differential gene expression tables and scatterplots. SPARTA provides an easy-to-use bacterial RNA-seq transcriptional profiling workflow to identify differentially expressed genes between experimental conditions. This software will enable microbiologists with

Surface charge-conversion polymeric nanoparticles for photodynamic treatment of urinary tract bacterial infections

NASA Astrophysics Data System (ADS)

Liu, Shijie; Qiao, Shenglin; Li, Lili; Qi, Guobin; Lin, Yaoxin; Qiao, Zengying; Wang, Hao; Shao, Chen

2015-12-01

Urinary tract infections are typical bacterial infections which result in a number of economic burdens. With increasing antibiotic resistance, it is urgent that new approaches are explored that can eliminate pathogenic bacteria without inducing drug resistance. Antimicrobial photodynamic therapy (PDT) is a new promising tactic. It is a gentle in situ photochemical reaction in which a photosensitizer (PS) generates reactive oxygen species (ROS) under laser irradiation. In this work, we have demonstrated Chlorin e6 (Ce6) encapsulated charge-conversion polymeric nanoparticles (NPs) for efficiently targeting and killing pathogenic bacteria in a weakly acidic urinary tract infection environment. Owing to the surface charge conversion of NPs in an acidic environment, the NPs exhibited enhanced recognition for Gram-positive (ex. S. aureus) and Gram-negative (ex. E. coli) bacteria due to the charge interaction. Also, those NPs showed significant antibacterial efficacy in vitro with low cytotoxicity. The MIC value of NPs to E. coli is 17.91 μg ml-1, compared with the free Ce6 value of 29.85 μg ml-1. Finally, a mouse acute cystitis model was used to assess the photodynamic therapy effects in urinary tract infections. A significant decline (P < 0.05) in bacterial cells between NPs and free Ce6 occurred in urine after photodynamic therapy treatment. And the plated counting results revealed a remarkable bacterial cells drop (P < 0.05) in the sacrificed bladder tissue. Above all, this nanotechnology strategy opens a new door for the treatment of urinary tract infections with minimal side effects.

Surface charge-conversion polymeric nanoparticles for photodynamic treatment of urinary tract bacterial infections.

PubMed

Liu, Shijie; Qiao, Shenglin; Li, Lili; Qi, Guobin; Lin, Yaoxin; Qiao, Zengying; Wang, Hao; Shao, Chen

2015-12-11

Urinary tract infections are typical bacterial infections which result in a number of economic burdens. With increasing antibiotic resistance, it is urgent that new approaches are explored that can eliminate pathogenic bacteria without inducing drug resistance. Antimicrobial photodynamic therapy (PDT) is a new promising tactic. It is a gentle in situ photochemical reaction in which a photosensitizer (PS) generates reactive oxygen species (ROS) under laser irradiation. In this work, we have demonstrated Chlorin e6 (Ce6) encapsulated charge-conversion polymeric nanoparticles (NPs) for efficiently targeting and killing pathogenic bacteria in a weakly acidic urinary tract infection environment. Owing to the surface charge conversion of NPs in an acidic environment, the NPs exhibited enhanced recognition for Gram-positive (ex. S. aureus) and Gram-negative (ex. E. coli) bacteria due to the charge interaction. Also, those NPs showed significant antibacterial efficacy in vitro with low cytotoxicity. The MIC value of NPs to E. coli is 17.91 μg ml(-1), compared with the free Ce6 value of 29.85 μg ml(-1). Finally, a mouse acute cystitis model was used to assess the photodynamic therapy effects in urinary tract infections. A significant decline (P < 0.05) in bacterial cells between NPs and free Ce6 occurred in urine after photodynamic therapy treatment. And the plated counting results revealed a remarkable bacterial cells drop (P < 0.05) in the sacrificed bladder tissue. Above all, this nanotechnology strategy opens a new door for the treatment of urinary tract infections with minimal side effects.

Relationship between salivary flow rates and Candida counts in subjects with xerostomia.

PubMed

Torres, Sandra R; Peixoto, Camila Bernardo; Caldas, Daniele Manhães; Silva, Eline Barboza; Akiti, Tiyomi; Nucci, Márcio; de Uzeda, Milton

2002-02-01

This study evaluated the relationship between salivary flow and Candida colony counts in the saliva of patients with xerostomia. Sialometry and Candida colony-forming unit (CFU) counts were taken from 112 subjects who reported xerostomia in a questionnaire. Chewing-stimulated whole saliva was collected and streaked in Candida plates and counted in 72 hours. Species identification was accomplished under standard methods. There was a significant inverse relationship between salivary flow and Candida CFU counts (P =.007) when subjects with high colony counts were analyzed (cutoff point of 400 or greater CFU/mL). In addition, the median sialometry of men was significantly greater than that of women (P =.003), even after controlling for confounding variables like underlying disease and medications. Sjögren's syndrome was associated with low salivary flow rate (P =.007). There was no relationship between the median Candida CFU counts and gender or age. There was a high frequency (28%) of mixed colonization. Candida albicans was the most frequent species, followed by C parapsilosis, C tropicalis, and C krusei. In subjects with high Candida CFU counts there was an inverse relationship between salivary flow and Candida CFU counts.

An evaluation of bacterial contamination of barriers used in periapical tissue regeneration: Part 2--Bacterial penetration.

PubMed

Sharma, Priya; Mickel, André K; Chogle, Sami; Sharma, Prem Nath; Han, Yiping W; Jones, Jefferson J

2008-03-01

To compare the relative penetration of Prevotella melaninogenica and Enterococcus faecalis through 3 guided tissue regeneration membranes: Atrisorb, Lambone, and OsseoQuest. It was hypothesized that OsseoQuest would show increased bacterial penetration when compared to Lambone and Atrisorb. Centrifuge tubes containing trypticase soy broth were sealed with circular sections of membranes and placed in test tubes containing culture media. The bacterial penetration was assessed by passage of bacteria from the outer tube culture media to the inner centrifuge tube media through the membrane. After incubation for 4 and 48 hours, the media from the outer and inner tubes were compared for bacterial count. P melaninogenica exhibited 91% penetration for Lambone in 2 days, while OsseoQuest displayed 87% penetration with E faecalis in the same time. Atrisorb displayed a minimal penetration with both bacteria (2%). Atrisorb displayed the least bacterial penetration, which may be attributed to membrane structure, chemical configuration, hydrophobicity, and porosity of tested membranes.

Multiple Heavy Metal Tolerance of Soil Bacterial Communities and Its Measurement by a Thymidine Incorporation Technique

PubMed Central

Díaz-Raviña, Montserrat; Bååth, Erland; Frostegård, Åsa

1994-01-01

A thymidine incorporation technique was used to determine the tolerance of a soil bacterial community to Cu, Cd, Zn, Ni, and Pb. An agricultural soil was artificially contaminated in our laboratory with individual metals at three different concentrations, and the results were compared with the results obtained by using the plate count technique. Thymidine incorporation was found to be a simple and rapid method for measuring tolerance. Data obtained by this technique were very reproducible. A linear relationship was found between changes in community tolerance levels obtained by the thymidine incorporation and plate count techniques (r = 0.732, P < 0.001). An increase in tolerance to the metal added to soil was observed for the bacterial community obtained from each polluted soil compared with the community obtained from unpolluted soil. The only exception was when Pb was added; no indication of Pb tolerance was found. An increase in the tolerance to metals other than the metal originally added to soil was also observed, indicating that there was multiple heavy metal tolerance at the community level. Thus, Cu pollution, in addition to increasing tolerance to Cu, also induced tolerance to Zn, Cd, and Ni. Zn and Cd pollution increased community tolerance to all five metals. Ni amendment increased tolerance to Ni the most but also increased community tolerance to Zn and, to lesser degrees, increased community tolerance to Pb and Cd. In soils polluted with Pb increased tolerance to other metals was found in the following order: Ni > Cd > Zn > Cu. We found significant positive relationships between changes in Cd, Zn, and Pb tolerance and, to a lesser degree, between changes in Pb and Ni tolerance when all metals and amendment levels were compared. The magnitude of the increase in heavy metal tolerance was found to be linearly related to the logarithm of the metal concentration added to the soil. Threshold tolerance concentrations were estimated from these linear

Bacterial adenosine triphosphate as a measure of urinary tract infection

NASA Technical Reports Server (NTRS)

Chappelle, E. W.; Picciolo, G. L.

1971-01-01

Procedure detects and counts bacteria present in urine samples. Method also determines bacterial levels in other aqueous body fluids including lymph fluid, plasma, blood, spinal fluid, saliva and mucous.

Long-term clinical and bacterial effects of xylitol on patients with fixed orthodontic appliances.

PubMed

Masoud, Mohamed I; Allarakia, Reem; Alamoudi, Najlaa M; Nalliah, Romesh; Allareddy, Veerasathpurush

2015-01-01

The objective of this study was to evaluate long-term clinical and bacterial effects of using 6 g of xylitol per day for 3 months on patients with full fixed orthodontic appliances. The study was a pilot clinical trial that included 41 subjects who were undergoing orthodontic treatment. The subjects were randomly divided into three groups. Group A received xylitol chewing gum, group B received xylitol dissolvable chewable tablets, and Group C served as the control group and did not receive xylitol gums or tablets. Clinical examination and the collection of plaque and saliva samples were carried out at baseline and 3, 6, and 12 months. All three groups were given oral hygiene instruction and were put on a 6-month cleaning and topical fluoride schedule. Plaque scores and bacterial counts were used to evaluate the effectiveness of the different approaches at reducing the caries risk. Xylitol groups did not experience any more reduction in plaque score, plaque MS counts, or salivary MS counts than the control group nor did they have lower values at any of the time points. Chewing gum did not significantly increase the incidence of debonded brackets over the other groups. Xylitol does not have a clinical or bacterial benefit in patients with fixed orthodontic appliances. Oral hygiene instructions and 6-month topical fluoride application were effective at reducing plaque scores and bacterial counts in patients with full fixed appliances regardless of whether or not xylitol was used.

Altered Virome and Bacterial Microbiome in Human Immunodeficiency Virus-Associated Acquired Immunodeficiency Syndrome

PubMed Central

Monaco, Cynthia L.; Gootenberg, David B.; Zhao, Guoyan; Handley, Scott A.; Ghebremichael, Musie S.; Lim, Efrem S.; Lankowski, Alex; Baldridge, Megan T.; Wilen, Craig B.; Flagg, Meaghan; Norman, Jason M.; Keller, Brian C.; Luévano, Jesús Mario; Wang, David; Boum, Yap; Martin, Jeffrey N.; Hunt, Peter W.; Bangsberg, David R.; Siedner, Mark J.; Kwon, Douglas S.; Virgin, Herbert W.

2016-01-01

SUMMARY Human immunodeficiency virus (HIV) infection is associated with increased intestinal translocation of microbial products and enteropathy as well as alterations in gut bacterial communities. However, whether the enteric virome contributes to this infection and resulting immunodeficiency remains unknown. We characterized the enteric virome and bacterial microbiome in a cohort of Ugandan patients, including HIV-uninfected or HIV-infected subjects and those either treated with anti-retroviral therapy (ART) or untreated. Low peripheral CD4 T cell counts were associated with an expansion of enteric adenovirus sequences and this increase was independent of ART treatment. Additionally, the enteric bacterial microbiome of patients with lower CD4 T counts exhibited reduced phylogenetic diversity and richness with specific bacteria showing differential abundance, including increases in Enterobacteriaceae, which have been associated with inflammation. Thus, immunodeficiency in progressive HIV infection is associated with alterations in the enteric virome and bacterial microbiome, which may contribute to AIDS-associated enteropathy and disease progression. PMID:26962942

Altered Virome and Bacterial Microbiome in Human Immunodeficiency Virus-Associated Acquired Immunodeficiency Syndrome.

PubMed

Monaco, Cynthia L; Gootenberg, David B; Zhao, Guoyan; Handley, Scott A; Ghebremichael, Musie S; Lim, Efrem S; Lankowski, Alex; Baldridge, Megan T; Wilen, Craig B; Flagg, Meaghan; Norman, Jason M; Keller, Brian C; Luévano, Jesús Mario; Wang, David; Boum, Yap; Martin, Jeffrey N; Hunt, Peter W; Bangsberg, David R; Siedner, Mark J; Kwon, Douglas S; Virgin, Herbert W

2016-03-09

Human immunodeficiency virus (HIV) infection is associated with increased intestinal translocation of microbial products and enteropathy as well as alterations in gut bacterial communities. However, whether the enteric virome contributes to this infection and resulting immunodeficiency remains unknown. We characterized the enteric virome and bacterial microbiome in a cohort of Ugandan patients, including HIV-uninfected or HIV-infected subjects and those either treated with anti-retroviral therapy (ART) or untreated. Low peripheral CD4 T cell counts were associated with an expansion of enteric adenovirus sequences and this increase was independent of ART treatment. Additionally, the enteric bacterial microbiome of patients with lower CD4 T counts exhibited reduced phylogenetic diversity and richness with specific bacteria showing differential abundance, including increases in Enterobacteriaceae, which have been associated with inflammation. Thus, immunodeficiency in progressive HIV infection is associated with alterations in the enteric virome and bacterial microbiome, which may contribute to AIDS-associated enteropathy and disease progression. Copyright © 2016 Elsevier Inc. All rights reserved.

Evaluation of the Microbiological Status of Raw Beef in Korea: Considering the Suitability of Aerobic Plate Count Guidelines

PubMed Central

Kim, Hye-Jin; Kim, Dongwook; Kim, Hee-Jin; Song, Sung-Ok; Song, Young-Han; Jang, Aera

2018-01-01

This study was conducted to analyze the microbiological contamination status of raw beef distributed in Korea, and evaluate the suitability of current aerobic plate count (APC) guidelines. We analyzed five years (2010-2014) of microbiological monitoring data obtained from the Ministry of Food and Drug Safety and investigated the microbiological status of raw beef collected from meat packing centers and meat shops in the Seoul/Gyeonggi, Gangwon, and Chungcheong regions in August 2015. From 2010-2014, most raw beef (>94%) displayed APC levels of < 1.0 × 106 CFU/g. However, raw beef samples collected from all three regions in August 2015 had comparatively higher APC levels than those reported in previous years. To evaluate the relationship between the APC level and quality, changes in beef loin were evaluated during cold storage for 15 days at 4°C. On day 11, the mean APC level (4.7 × 106 CFU/g) conformed to current guidelines in Korea (1.0 × 107 CFU/g) and the pH value was 5.82. However, the sensory evaluation score for color and overall acceptability was under 3.0, meaning that the beef loin was not acceptable for eating. These results suggest that current APC guideline for raw beef should be lowered to 1.0 × 106 CFU/g to improve both the microbiological safety and palatability of raw beef. PMID:29725223

Transit time affects the community stability of Lactobacillus and Bifidobacterium species in an in vitro model of human colonic microbiotia.

PubMed

Rodes, Laetitia; Paul, Arghya; Coussa-Charley, Michael; Al-Salami, Hani; Tomaro-Duchesneau, Catherine; Fakhoury, Marc; Prakash, Satya

2011-12-01

Retention time, which is analogous to transit time, is an index for bacterial stability in the intestine. Its consideration is of particular importance to optimize the delivery of probiotic bacteria in order to improve treatment efficacy. This study aims to investigate the effect of retention time on Lactobacilli and Bifidobacteria stability using an established in vitro human colon model. Three retention times were used: 72, 96, and 144 h. The effect of retention time on cell viability of different bacterial populations was analyzed with bacterial plate counts and PCR. The proportions of intestinal Bifidobacteria, Lactobacilli, Enterococci, Staphylococci and Clostridia populations, analyzed by plate counts, were found to be the same as that in human colonic microbiota. Retention time in the human colon affected the stability of Lactobacilli and Bifidobacteria communities, with maximum stability observed at 144 h. Therefore, retention time is an important parameter that influences bacterial stability in the colonic microbiota. Future clinical studies on probiotic bacteria formulations should take into consideration gastrointestinal transit parameters to improve treatment efficacy.

Non-homogeneous flow profiles in sheared bacterial suspensions

NASA Astrophysics Data System (ADS)

Samanta, Devranjan; Cheng, Xiang

Bacterial suspensions under shear exhibit interesting rheological behaviors including the remarkable ``superfluidic'' state with vanishing viscosity at low shear rates. Theoretical studies have shown that such ``superfluidic'' state is linked with non-homogeneous shear flows, which are induced by coupling between nematic order of active fluids and hydrodynamics of shear flows. However, although bulk rheology of bacterial suspensions has been experimentally studied, shear profiles within bacterial suspensions have not been explored so far. Here, we experimentally investigate the flow behaviors of E. coli suspensions under planar oscillatory shear. Using confocal microscopy and PIV, we measure velocity profiles across gap between two shear plates. We find that with increasing shear rates, high-concentration bacterial suspensions exhibit an array of non-homogeneous flow behaviors like yield-stress flows and shear banding. We show that these non-homogeneous flows are due to collective motion of bacterial suspensions. The phase diagram of sheared bacterial suspensions is systematically mapped as functions of shear rates an bacterial concentrations. Our experiments provide new insights into rheology of bacterial suspensions and shed light on shear induced dynamics of active fluids. Chemical Engineering and Material Science department.

High event rate ROICs (HEROICs) for astronomical UV photon counting detectors

NASA Astrophysics Data System (ADS)

Harwit, Alex; France, Kevin; Argabright, Vic; Franka, Steve; Freymiller, Ed; Ebbets, Dennis

2014-07-01

The next generation of astronomical photocathode / microchannel plate based UV photon counting detectors will overcome existing count rate limitations by replacing the anode arrays and external cabled electronics with anode arrays integrated into imaging Read Out Integrated Circuits (ROICs). We have fabricated a High Event Rate ROIC (HEROIC) consisting of a 32 by 32 array of 55 μm square pixels on a 60 μm pitch. The pixel sensitivity (threshold) has been designed to be globally programmable between 1 × 103 and 1 × 106 electrons. To achieve the sensitivity of 1 × 103 electrons, parasitic capacitances had to be minimized and this was achieved by fabricating the ROIC in a 65 nm CMOS process. The ROIC has been designed to support pixel counts up to 4096 events per integration period at rates up to 1 MHz per pixel. Integration time periods can be controlled via an external signal with a time resolution of less than 1 microsecond enabling temporally resolved imaging and spectroscopy of astronomical sources. An electrical injection port is provided to verify functionality and performance of each ROIC prior to vacuum integration with a photocathode and microchannel plate amplifier. Test results on the first ROICs using the electrical injection port demonstrate sensitivities between 3 × 103 and 4 × 105 electrons are achieved. A number of fixes are identified for a re-spin of this ROIC.

The use of multiple indices of physiological activity to access viability in chlorine disinfected Escherichia coli O157:H7

NASA Technical Reports Server (NTRS)

Lisle, J. T.; Pyle, B. H.; McFeters, G. A.

1999-01-01

A suite of fluorescent intracellular stains and probes was used, in conjunction with viable plate counts, to assess the effect of chlorine disinfection on membrane potential (rhodamine 123; Rh123 and bis-(1,3-dibutylbarbituric acid) trimethine oxonol; DiBAC4(3)), membrane integrity (LIVE/DEAD BacLight kit), respiratory activity (5-cyano-2,3-ditolyl tetrazolium chloride; CTC) and substrate responsiveness (direct viable counts; DVC) in the commensal pathogen Escherichia coli O157:H7. After a 5 min exposure to the disinfectant, physiological indices were affected in the following order: viable plate counts > substrate responsiveness > membrane potential > respiratory activity > membrane integrity. In situ assessment of physiological activity by examining multiple targets, as demonstrated in this study, permits a more comprehensive determination of the site and extent of injury in bacterial cells following sublethal disinfection with chlorine. This approach to assessing altered bacterial physiology has application in various fields where detection of stressed bacteria is of interest.

Dynamic light scattering: A fast and reliable method to analyze bacterial growth during the lag phase.

PubMed

Vargas, Susana; Millán-Chiu, Blanca E; Arvizu-Medrano, Sofía M; Loske, Achim M; Rodríguez, Rogelio

2017-06-01

A comparison between plate counting (PC) and dynamic light scattering (DLS) is reported. PC is the standard technique to determine bacterial population as a function of time; however, this method has drawbacks, such as the cumbersome preparation and handling of samples, as well as the long time required to obtain results. Alternative methods based on optical density are faster, but do not distinguish viable from non-viable cells. These inconveniences are overcome by using DLS. Two different bacteria strains were considered: Escherichia coli and Staphylococcus aureus. DLS was performed at two different illuminating conditions: continuous and intermittent. By the increment of particle size as a function of time, it was possible to observe cell division and the formation of aggregates containing very few bacteria. The scattered intensity profiles showed the lag phase and the transition to the exponential phase of growth, providing a quantity proportional to viable bacteria concentration. The results revealed a clear and linear correlation in both lag and exponential phase, between the Log 10 (colony-forming units/mL) from PC and the Log 10 of the scattered intensity I s from DLS. These correlations provide a good support to use DLS as an alternative technique to determine bacterial population. Copyright © 2017 Elsevier B.V. All rights reserved.

Validation of the bacterial meningitis score in adults presenting to the ED with meningitis.

PubMed

McArthur, Robert; Edlow, Jonathan A; Nigrovic, Lise E

2016-07-01

The Bacterial Meningitis Score classifies children with meningitis and none of the following high-risk predictors at very low risk for bacterial meningitis: positive cerebrospinal fluid (CSF) Gram stain, CSF protein ≥80mg/dL, CSF absolute neutrophil count (ANC) ≥1000 cells/mm(3), peripheral ANC ≥10,000 cells/mm(3), and seizure at or prior to presentation. Although extensively validated in children, the Bacterial Meningitis Score has not been rigorously evaluated in adults. We performed a single-center cross-sectional retrospective study of adults presenting to the emergency department between 2003 and 2013 with meningitis (defined by CSF white blood cell count ≥10 cells/mm(3)). We defined a case of bacterial meningitis with either a positive CSF or blood culture. We report the performance of the Bacterial Meningitis Score in the study population. We identified 441 eligible patients of which, 4 (1%) had bacterial meningitis. The Bacterial Meningitis Score had a sensitivity of 100% [95% confidence interval (CI) 40%-100%], specificity 51% (95% CI, 46%-56%) and negative predictive value of 100% (95% CI, 98%-100%). None of the low risk adults had bacterial meningitis. If Bacterial Meningitis Score had been applied prospectively, the hospital admission rate would have dropped from 84% to 49% without missing any patients with bacterial meningitis. The Bacterial Meningitis Score accurately identified patients at low risk for bacterial meningitis and could assist clinical decision-making for adults with meningitis. Copyright © 2016 Elsevier Inc. All rights reserved.

TEMPERATURE-GRADIENT PLATES FOR GROWTH OF MICROORGANISMS

PubMed Central

Landman, Otto E.; Bausum, Howard T.; Matney, Thomas S.

1962-01-01

Landman, Otto E. (Fort Detrick, Frederick, Md.), Howard T. Bausum, and Thomas S. Matney. Temperature-gradient plates for growth of microorganisms. J. Bacteriol. 83:463–469. 1962.—Different temperature-gradient plates have been devised for the study of microbial growth on solid media through continuous temperature ranges or in liquid media at finely graded temperatures. All plates are made of heavy-gauge aluminum; heat supplied at one end is dissipated along the length of the metal so that a gradient is produced. The shape and range of the gradient depends on the amount of heat supplied, the insulation, the ambient temperature, and other factors. Differences of 0.2 C in temperature sensitivity between bacterial strains can be detected. The plates are simple to construct and operate. The dimensions of the aluminum, the mode of temperature measurement, and the method of heating may all be modified without diminishing the basic utility of the device. A sharp growth front develops at the maximal temperature of growth of bacteria. In most strains, all bacteria below the front form colonies and all bacteria above the front are killed, except for a few temperature-resistant mutants. Images PMID:14461975

Bacterial desorption from food container and food processing surfaces.

PubMed

McEldowney, S; Fletcher, M

1988-03-01

The desorption ofStaphylococcus aureus, Acinetobacter calcoaceticus, and a coryneform from the surfaces of materials used for manufacturing food containers (glass, tin plate, and polypropylene) or postprocess canning factory conveyor belts (stainless steel and nylon) was investigated. The effect of time, pH, temperature, and adsorbed organic layers on desorption was studied.S. aureus did not detach from the substrata at any pH investigated (between pH 5 and 9).A. calcoaceticus and the coryneform in some cases detached, depending upon pH and substratum composition. The degree of bacterial detachment from the substrata was not related to bacterial respiration at experimental pH values. Bacterial desorption was not affected by temperature (4-30°C) nor by an adsorbed layer of peptone and yeast extract on the substrata. The results indicate that bacterial desorption, hence bacterial removal during cleaning or their transfer via liquids flowing over colonized surfaces, is likely to vary with the surface composition and the bacterial species colonizing the surfaces.

Mobile laminar air flow screen for additional operating room ventilation: reduction of intraoperative bacterial contamination during total knee arthroplasty.

PubMed

Sossai, D; Dagnino, G; Sanguineti, F; Franchin, F

2011-12-01

Surgical site infections are important complications in orthopedic surgery. A mobile laminar air flow (LAF) screen could represent a useful addition to an operating room (OR) with conventional turbulent air ventilation (12.5 air changes/h), as it could decrease the bacterial count near the operating field. The purpose of this study was to evaluate LAF efficacy at reducing bacterial contamination in the surgical area during 34 total knee arthroplasties (TKAs). The additional unit was used in 17 operations; the LAF was positioned beside the operating table between two of the surgeons, with the air flow directed towards the surgical area (wound). The whole team wore conventional OR clothing and the correct hygiene procedures and rituals were used. Bacterial air contamination (CFU/m(3)) was evaluated in the wound area in 17 operations with the LAF unit and 17 without the LAF unit. The LAF unit reduced the mean bacterial count in the wound area from 23.5 CFU/m(3) without the LAF to 3.5 CFU/m(3) with the LAF (P < 0.0001), which is below the suggested limit for an OR with ultraclean laminar ventilation. There were no significant differences in the mean bacterial count in the instrument table area: 28.6 CFU/m(3) were recorded with the LAF (N = 6) unit and 30.8 CFU/m(3) (N = 6) without the LAF unit (P = 0.631). During six operations with LAF and six without LAF, particle counts were performed and the number of 0.5 μm particles was analyzed. The particle counts decreased significantly when the LAF unit was used (P = 0.003). When a mobile LAF unit was added to the standard OR ventilation, bacterial contamination of the wound area significantly decreased to below the accepted level for an ultraclean OR, preventing SSI infections.

Changes in bacterial diversity associated with bioremediation of used lubricating oil in tropical soils.

PubMed

Meeboon, Naruemon; Leewis, Mary-Cathrine; Kaewsuwan, Sireewan; Maneerat, Suppasil; Leigh, Mary Beth

2017-08-01

Used lubricating oil (ULO) is a widespread contaminant, particularly throughout tropical regions, and may be a candidate for bioremediation. However, little is known about the biodegradation potential or basic microbial ecology of ULO-contaminated soils. This study aims to determine the effects of used ULO on bacterial community structure and diversity. Using a combination of culture-based (agar plate counts) and molecular techniques (16S rRNA gene sequencing and DGGE), we investigated changes in soil bacterial communities from three different ULO-contaminated soils collected from motorcycle mechanical workshops (soil A, B, and C). We further explored the relationship between bacterial community structure, physiochemical soil parameters, and ULO composition in three ULO-contaminated soils. Results indicated that the three investigated soils had different community structures, which may be a result of the different ULO characteristics and physiochemical soil parameters of each site. Soil C had the highest ULO concentration and also the greatest diversity and richness of bacteria, which may be a result of higher nutrient retention, organic matter and cation exchange capacity, as well as freshness of oil compared to the other soils. In soils A and B, Proteobacteria (esp. Gammaproteobacteria) dominated the bacterial community, and in soil C, Actinobacteria and Firmicutes dominated. The genus Enterobacter, a member of the class Gammaproteobacteria, is known to include ULO-degraders, and this genus was the only one found in all three soils, suggesting that it could play a key role in the in situ degradation of ULO-contaminated tropical Thai soils. This study provides insights into our understanding of soil microbial richness, diversity, composition, and structure in tropical ULO-contaminated soils, and may be useful for the development of strategies to improve bioremediation.

Analysis of clinical outcomes in pediatric bacterial meningitis focusing on patients without cerebrospinal fluid pleocytosis.

PubMed

Lin, Wen-Li; Chi, Hsin; Huang, Fu-Yuan; Huang, Daniel Tsung-Ning; Chiu, Nan-Chang

2016-10-01

Cerebrospinal fluid (CSF) cell count and biochemical examinations and cultures form the basis for the diagnosis of bacterial meningitis. However, some patients do not have typical findings and are at a higher risk of being missed or having delayed treatment. To better understand the correlation between CSF results and outcomes, we evaluated CSF data focusing on the patients with atypical findings. This study enrolled CSF culture-proven bacterial meningitis patients aged from 1 month to 18 years in a medical center. The patients were divided into "normal" and "abnormal" groups for each laboratory result and in combination. The correlations between the laboratory results and the outcomes were analyzed. A total of 175 children with confirmed bacterial meningitis were enrolled. In CSF examinations, 16.2% of patients had normal white blood cell counts, 29.5% had normal glucose levels, 24.5% had normal protein levels, 10.2% had normal results in two items, and 8.6% had normal results in all three items. In logistic regression analysis, a normal CSF leukocyte count and increased CSF protein level were related to poor outcomes. Patients with meningitis caused by Streptococcus pneumoniae and hyponatremia were at a higher risk of mortality and the development of sequelae. In children with bacterial meningitis, nontypical CSF findings and, in particular, normal CSF leukocyte count and increased protein level may indicate a worse prognosis. Copyright © 2014. Published by Elsevier B.V.

Machine for Automatic Bacteriological Pour Plate Preparation

PubMed Central

Sharpe, A. N.; Biggs, D. R.; Oliver, R. J.

1972-01-01

A fully automatic system for preparing poured plates for bacteriological analyses has been constructed and tested. The machine can make decimal dilutions of bacterial suspensions, dispense measured amounts into petri dishes, add molten agar, mix the dish contents, and label the dishes with sample and dilution numbers at the rate of 2,000 dishes per 8-hr day. In addition, the machine can be programmed to select different media so that plates for different types of bacteriological analysis may be made automatically from the same sample. The machine uses only the components of the media and sterile polystyrene petri dishes; requirements for all other materials, such as sterile pipettes and capped bottles of diluents and agar, are eliminated. Images PMID:4560475

Effects of dietary inclusion of silymarin on performance, intestinal morphology and ileal bacterial count in aflatoxin-challenged broiler chicks.

PubMed

Jahanian, E; Mahdavi, A H; Asgary, S; Jahanian, R

2017-10-01

This study was conducted to investigate the effect of dietary supplementation of silymarin on performance, jejunal morphology and ileal bacterial population in broiler chicks intoxicated with a mix of aflatoxins. A total of three hundred thirty six 7-day-old Ross broiler chicks were randomly distributed between seven experimental groups with four replicates of 12 birds each. Experimental treatments consisted of a control group (unchallenged), and a 2 × 3 factorial arrangement, including two aflatoxin levels (0.5 and 2 ppm) and three levels of silymarin (0, 500 and 1000 ppm). Birds were challenged with a mix of aflatoxins from 7 to 28 days of age. Results showed that increasing aflatoxin level resulted in decreased average daily feed intake (ADFI) and weight gain (ADWG), consequently impaired feed conversion ratio (FCR) throughout the trial period. Dietary supplementation of silymarin resulted in the marked increases in ADFI and ADWG, and improved FCR values in aflatoxin-challenged chicks. Ileal bacterial populations at days 28 and 42 of age were increased by incremental levels of aflatoxins. On the other hand, dietary silymarin supplementation suppressed ileal populations of Escherichia coli, Salmonella, Klebsiella and total negative bacteria in aflatoxicated birds. Increase in dietary aflatoxin level resulted in the decreased villi height, villi height-to-crypt depth ratio (VH:CD), villi surface area and apparent villi absorptive area, while it increased crypt depth, goblet cell count and lymphoid follicular diameter. Feeding silymarin at the level of 1000 ppm increased villi height and VH:CD in aflatoxicated birds. Present results indicate that dietary inclusion of silymarin could improve performance by suppressing ileal bacteria and enhancing absorptive surface area in aflatoxin-challenged broiler chicks. Journal of Animal Physiology and Animal Nutrition © 2017 Blackwell Verlag GmbH.

Portable bacterial identification system based on elastic light scatter patterns.

PubMed

Bae, Euiwon; Ying, Dawei; Kramer, Donald; Patsekin, Valery; Rajwa, Bartek; Holdman, Cheryl; Sturgis, Jennifer; Davisson, V Jo; Robinson, J Paul

2012-08-28

Conventional diagnosis and identification of bacteria requires shipment of samples to a laboratory for genetic and biochemical analysis. This process can take days and imposes significant delay to action in situations where timely intervention can save lives and reduce associated costs. To enable faster response to an outbreak, a low-cost, small-footprint, portable microbial-identification instrument using forward scatterometry has been developed. This device, weighing 9 lb and measuring 12 × 6 × 10.5 in., utilizes elastic light scatter (ELS) patterns to accurately capture bacterial colony characteristics and delivers the classification results via wireless access. The overall system consists of two CCD cameras, one rotational and one translational stage, and a 635-nm laser diode. Various software algorithms such as Hough transform, 2-D geometric moments, and the traveling salesman problem (TSP) have been implemented to provide colony count and circularity, centering process, and minimized travel time among colonies. Experiments were conducted with four bacteria genera using pure and mixed plate and as proof of principle a field test was conducted in four different locations where the average classification rate ranged between 95 and 100%.

Microchannel plate life testing for UV spectroscopy instruments

NASA Astrophysics Data System (ADS)

Darling, N. T.; Siegmund, O. H. W.; Curtis, T.; McPhate, J.; Tedesco, J.; Courtade, S.; Holsclaw, G.; Hoskins, A.; Al Dhafri, S.

2017-08-01

The Emirates Mars Mission (EMM) UV Spectrograph (EMUS) is a far ultraviolet (102 nm to 170 nm) imaging spectrograph for characterization of the Martian exosphere and thermosphere. Imaging is accomplished by a photon counting open-face microchannel plate (MCP) detector using a cross delay line (XDL) readout. An MCP gain stabilization ("scrub") followed by lifetime spectral line burn-in simulation has been completed on a bare MCP detector at SSL. Gain and sensitivity stability of better than 7% has been demonstrated for total dose of 2.5 × 1012 photons cm-2 (2 C · cm-2 ) at 5.5 kHz mm-2 counting rates, validating the efficacy of an initial low gain full-field scrub.

Microchannel plate EUV detectors for the Extreme Ultraviolet Explorer

NASA Technical Reports Server (NTRS)

Siegmund, O. H. W.; Malina, R. F.; Coburn, K.; Werthimer, D.

1984-01-01

The design and operating characteristics of the prototype imaging microchannel plate (MCP) detector for the Extreme Ultraviolet Explorer (EUVE) Satellite are discussed. It is shown that this detector has achieved high position resolution performance (greater than 512 x 512 pixels) and has low (less than one percent) image distortion. In addition, the channel plate scheme used has tight pulse height distributions (less than 40 percent FWHM) for UV radiation and displays low (less than 0.2 cnt/sq cm-s) dark background counting rates. Work that has been done on EUV filters in relation to the envisaged filter and photocathode complement is also described.

An evidential example of airborne bacteria in a crowded, underground public concourse in Tokyo

NASA Astrophysics Data System (ADS)

Seino, Kaoruko; Takano, Takehito; Nakamura, Keiko; Watanabe, Masafumi

2005-01-01

We examined airborne bacteria in an underground concourse in Tokyo and investigated conditions that influenced bacterial counts. Airborne bacteria were collected by using an impactor sampler. Colonies on plate count agar (PCA) and Columbia colistin-nalidixic acid agar with 5% sheep blood (CNA agar) were enumerated. The range, geometric mean, and 95% CI of the bacterial counts (CFU m-3) on PCA and CNA agar were 150-1380, 456, 382-550 and 50-990, 237, 182-309, respectively. Bacterial counts on PCA significantly correlated with number of the pedestrians (r=0.89), relative humidity (r=0.70) and airborne dust (PM5.0) (r=0.73). Results of a multiple regression indicated independent positive association between the number of pedestrians and bacterial counts on PCA (p<0.01) after excluding the influence of relative humidity and airborne dust. Similar results were obtained with the statistical analysis for the counts of bacteria on CNA agar. Gram-positive cocci were dominant on PCA and CNA agar. Staphylococcus epidermidis and Micrococcus spp. were dominant among the 11 genera and 19 species identified in the present study. Considering the pattern of identified species and the significant independent association between number of pedestrians and bacterial counts, airborne bacteria in a crowded underground concourse were mostly originated from the pedestrians who were walking in the underground concourse. This study gave an evidential example of bacterial conditions in the air of an underground crowded public space in Tokyo.

Plasma surface modification of rigid contact lenses decreases bacterial adhesion.

PubMed

Wang, Yingming; Qian, Xuefeng; Zhang, Xiaofeng; Xia, Wei; Zhong, Lei; Sun, Zhengtai; Xia, Jing

2013-11-01

Contact lens safety is an important topic in clinical studies. Corneal infections usually occur because of the use of bacteria-carrying contact lenses. The current study investigated the impact of plasma surface modification on bacterial adherence to rigid contact lenses made of fluorosilicone acrylate materials. Boston XO and XO2 contact lenses were modified using plasma technology (XO-P and XO2-P groups). Untreated lenses were used as controls. Plasma-treated and control lenses were incubated in solutions containing Staphylococcus aureus or Pseudomonas aeruginosa. MTT colorimetry, colony-forming unit counting method, and scanning electron microscopy were used to measure bacterial adhesion. MTT colorimetry measurements showed that the optical density (OD) values of XO-P and XO2-P were significantly lower than those of XO and XO2, respectively, after incubation with S. aureus (P < 0.01). The OD value of XO-P was also much lower than that of XO after incubation with P. aeruginosa (P < 0.01). Colony-forming unit counting revealed that a significantly lower number of bacterial colonies attached to the XO-P versus XO lenses and to the XO2-P versus XO2 lenses incubated with S. aureus (P < 0.01). Fewer bacterial colonies attached to the XO-P versus XO lenses incubated with P. aeruginosa (P < 0.01). Further, scanning electron microscopy suggested different bacterial adhesion morphology on plasma-treated versus control lenses. Plasma surface modification can significantly decrease bacterial adhesion to fluorosilicone acrylate contact lenses. This study provides important evidence of a unique benefit of plasma technology in contact lens surface modification.

Effect of human milk fortifiers on bacterial growth in human milk.

PubMed

Santiago, Myla S; Codipilly, Champa N; Potak, Debra C; Schanler, Richard J

2005-10-01

As a component in human milk fortifiers (HMF), iron may equilibrate with human milk for as long as 24 hours, bind important bacteriostatic proteins, and potentially affect the host defense properties of human milk. We compared bacterial growth in human milk prepared with each of two HMF differing in their content of iron. Samples of human milk obtained from mothers of premature infants were divided and mixed with one of two HMF and maintained at refrigerator temperature. Refrigerated milk samples were removed at 0, 24, and 72 hours for determination of total bacterial colony counts (TBCC). TBCC did not differ between groups but declined from 0 to 72 hours, p<0.001. These data suggest that differences in iron content, or other nutrients in HMF, do not affect bacterial growth in human milk. Storage of fortified human milk at refrigerator temperature for 72 hours results in decreased bacterial growth. As a component in human milk fortifiers (HMF), iron may equilibrate with human milk for as long as 24 hours, bind important bacteriostatic proteins, and potentially affect the host defense properties of human milk. We compared bacterial growth in human milk prepared with each of two HMF differing in their content of iron. Samples of human milk obtained from mothers of premature infants were divided and mixed with one of two HMF and maintained at refrigerator temperature. Refrigerated milk samples were removed at 0, 24, and 72 hours for determination of total bacterial colony counts (TBCC).

Survivial Strategies in Bacterial Range Expansions

NASA Astrophysics Data System (ADS)

Frey, Erwin

2014-03-01

Bacterial communities represent complex and dynamic ecological systems. Different environmental conditions as well as bacterial interactions determine the establishment and sustainability of bacterial diversity. In this talk we discuss the competition of three Escherichia coli strains during range expansions on agar plates. In this bacterial model system, a colicin E2 producing strain C competes with a colicin resistant strain R and with a colicin sensitive strain S for new territory. Genetic engineering allows us to tune the growth rates of the strains and to study distinct ecological scenarios. These scenarios may lead to either single-strain dominance, pairwise coexistence, or to the coexistence of all three strains. In order to elucidate the survival mechanisms of the individual strains, we also developed a stochastic agent-based model to capture the ecological scenarios in silico. In a combined theoretical and experimental approach we are able to show that the level of biodiversity depends crucially on the composition of the inoculum, on the relative growth rates of the three strains, and on the effective reach of colicin toxicity.

Induction of a global stress response during the first step of Escherichia coli plate growth.

PubMed

Cuny, Caroline; Lesbats, Maïalène; Dukan, Sam

2007-02-01

We have investigated the first events that occur when exponentially grown cells are transferred from a liquid medium (Luria-Bertani [LB]) to a solid medium (LB agar [LBA]). We observed an initial lag phase of 180 min for the wild type MG1655 without any apparent growth. This lack of growth was independent of the bacterial physiological state (either the stationary or the exponential phase), the solid medium composition, or the number of cells on the plate, but it was dependent on the bacterial genotype. Using lacZ-reporter fusions and two-dimensional electrophoresis analysis, we observed that when cells from exponential-phase cultures were plated on LBA, several global regulons, like heat shock regulons (RpoH, RpoE, CpxAR) and oxidative-stress regulons (SoxRS, OxyR, Fur), were immediately induced. Our results indicate that in order to grow on plates, bacteria must not only adapt to new conditions but also perceive a real stress.

Rapid depletion of dissolved oxygen in 96-well microtiter plate Staphylococcus epidermidis biofilm assays promotes biofilm development and is influenced by inoculum cell concentration.

PubMed

Cotter, John J; O'Gara, James P; Casey, Eoin

2009-08-01

Biofilm-related research using 96-well microtiter plates involves static incubation of plates indiscriminate of environmental conditions, making oxygen availability an important variable which has not been considered to date. By directly measuring dissolved oxygen concentration over time we report here that dissolved oxygen is rapidly consumed in Staphylococcus epidermidis biofilm cultures grown in 96-well plates irrespective of the oxygen concentration in the gaseous environment in which the plates are incubated. These data indicate that depletion of dissolved oxygen during growth of bacterial biofilm cultures in 96-well plates may significantly influence biofilm production. Furthermore higher inoculum cell concentrations are associated with more rapid consumption of dissolved oxygen and higher levels of S. epidermidis biofilm production. Our data reveal that oxygen depletion during bacterial growth in 96-well plates may significantly influence biofilm production and should be considered in the interpretation of experimental data using this biofilm model.

Non-thermal effects of 94 GHz radiation on bacterial metabolism

NASA Astrophysics Data System (ADS)

Raitt, Brittany J.

Bacillus subtilis, Escherichia coli, Staphylococcus aureus, and Klebsiella pneumoniae were used to investigate the non-thermal effects of terahertz (THz) radiation exposure on bacterial cells. The THz source used was a 94 GHz (0.94 THz) Millitech Gunn Diode Oscillator with a power density of 1.3 mW/cm2. The cultures were placed in the middle sixty wells of two 96-well microplates, one serving as the experimental plate and one serving as a control. The experimental plate was placed on the radiation source for either two, eighteen, or twenty-four hours and the metabolism of the cells was measured in a spectrophotometer using the tetrazolium dye XTT. The results showed no consistent significant differences in either the growth rates or the metabolism of any of the bacterial species at this frequency and power density.

Intermittent fasting promotes bacterial clearance and intestinal IgA production in Salmonella typhimurium-infected mice.

PubMed

Godínez-Victoria, M; Campos-Rodriguez, R; Rivera-Aguilar, V; Lara-Padilla, E; Pacheco-Yepez, J; Jarillo-Luna, R A; Drago-Serrano, M E

2014-05-01

The impact of intermittent fasting versus ad libitum feeding during Salmonella typhimurium infection was evaluated in terms of duodenum IgA levels, bacterial clearance and intestinal and extra-intestinal infection susceptibility. Mice that were intermittently fasted for 12 weeks or fed ad libitum were infected with S. typhimurium and assessed at 7 and 14 days post-infection. Next, we evaluated bacterial load in the faeces, Peyer's patches, spleen and liver by plate counting, as well as total and specific intestinal IgA and plasmatic corticosterone levels (by immunoenzymatic assay) and lamina propria IgA levels in plasma cells (by cytofluorometry). Polymeric immunoglobulin receptor, α- and J-chains, Pax-5 factor, pro-inflammatory cytokine (tumour necrosis factor-α and interferon-γ) and anti-inflammatory cytokine (transforming growth factor-β) mRNA levels were assessed in mucosal and liver samples (by real-time PCR). Compared with the infected ad libitum mice, the intermittently fasted infected animals had (1) lower intestinal and systemic bacterial loads; (2) higher SIgA and IgA plasma cell levels; (3) higher mRNA expression of most intestinal parameters; and (4) increased or decreased corticosterone levels on day 7 and 14 post-infection, respectively. No contribution of liver IgA was observed at the intestinal level. Apparently, the changes following metabolic stress induced by intermittent fasting during food deprivation days increased the resistance to S. typhimurium infection by triggering intestinal IgA production and presumably, pathogen elimination by phagocytic inflammatory cells. © 2014 John Wiley & Sons Ltd.

Dynamics of bacterial communities before and after distribution in a full-scale drinking water network.

PubMed

El-Chakhtoura, Joline; Prest, Emmanuelle; Saikaly, Pascal; van Loosdrecht, Mark; Hammes, Frederik; Vrouwenvelder, Hans

2015-05-01

Understanding the biological stability of drinking water distribution systems is imperative in the framework of process control and risk management. The objective of this research was to examine the dynamics of the bacterial community during drinking water distribution at high temporal resolution. Water samples (156 in total) were collected over short time-scales (minutes/hours/days) from the outlet of a treatment plant and a location in its corresponding distribution network. The drinking water is treated by biofiltration and disinfectant residuals are absent during distribution. The community was analyzed by 16S rRNA gene pyrosequencing and flow cytometry as well as conventional, culture-based methods. Despite a random dramatic event (detected with pyrosequencing and flow cytometry but not with plate counts), the bacterial community profile at the two locations did not vary significantly over time. A diverse core microbiome was shared between the two locations (58-65% of the taxa and 86-91% of the sequences) and found to be dependent on the treatment strategy. The bacterial community structure changed during distribution, with greater richness detected in the network and phyla such as Acidobacteria and Gemmatimonadetes becoming abundant. The rare taxa displayed the highest dynamicity, causing the major change during water distribution. This change did not have hygienic implications and is contingent on the sensitivity of the applied methods. The concept of biological stability therefore needs to be revised. Biostability is generally desired in drinking water guidelines but may be difficult to achieve in large-scale complex distribution systems that are inherently dynamic. Copyright © 2015 Elsevier Ltd. All rights reserved.

Enumeration of the contaminating bacterial microbiota in unfermented pasteurized milks enriched with probiotic bacteria.

PubMed

Champagne, C P; Raymond, Y; Gonthier, J; Audet, P

2009-04-01

Pasteurized and unfermented milks supplemented with probiotic bacteria are appearing on the market. It then becomes a challenge to ascertain the undesirable contamination microbiota in the presence of a largely superior population of probiotic bacteria. A method to enumerate the contaminating microbial microbiota in such probiotic-enriched milks was developed. The probiotic cultures, Lactobacillus rhamnosus Lb-Immuni-T and Bifidobacterium animalis subsp. lactis BB-12(R), were added to a pasteurized unfermented milk to reach a minimum of 1 billion CFU per 250 mL portion, as ascertained by plating on de Man - Rogosa - Sharpe (MRS) agar in anaerobic conditions. No growth of B. animalis subsp. lactis BB-12 was noted on plate count agar (PCA) or Petrifilm plates, and the presence of this culture did not affect standard plate counts (SPC) of contaminating bacteria. However, L. rhamnosus formed colonies on PCA and Petrifilm plates. Attempts were thus made to inhibit the growth of the probiotic lactobacilli in PCA. The addition of 2% sodium phosphate (SP) or 5% glycerophosphate (GP) inhibited the growth of the lactobacilli in broths, but pin-point colonies of L. rhamnosus Lb-Immuni-T nevertheless appeared on PCA supplemented with phosphates. SPC could be obtained on PCA + 2% SP by only counting the large colonies, but this resulted in a significant (4.4 fold) underestimation of SPC values. On Petrifilm AC, at dilutions 0 to 2, all colonies were considered as being contaminants, while at dilutions 3 and 4, only large colonies were counted for SPC determinations. There was a direct correlation (R2 = 0.99) between SPC values with Petrifilm in uninoculated milks and those obtained on probiotic-enriched milks. The high correlation obtained over the 102 to 106 CFU/mL range of SPC values show that this Petrifilm method is appropriate to evaluate the microbiological quality of pasteurized milks enriched with L. rhamnosus Lb-Immuni-T and B. animalis subsp. lactis BB-12.

[Analysis of bactericidal material generated by electrical devices advertising bactericidal ability against bacteria on the agar gel plates].

PubMed

Nishimura, Hidekazu

2012-11-01

Several Japanese companies sell electrical devices advertised as effective in inactivating viruses and killing bacteria by releasing special materials, e.g., Plasmacluster ions, Nanoe particle and minus ions, into the air. These companies claim that their devices killed bacteria on plates in their own experiments. We tested device effectiveness using the same experiments from the Plasmacluster ioniser SHARP Co., Japan, the Nanoe generator Panasonic Co., Japan, and the Vion KING JIM Co., Japan, to test their advertising claims. Bactericidal ability on agar plate was tested, using Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus cereus, and Enterococcus faecalis as follows: the medium containing a certain amount of each bacterium was put onto an agar plate and smeared. Plates were kept in a closed chamber (inner volume 14.4 m3) or a glove box (inner volume 0.2 m), with one of the devices run for 2 hours. Plates not exposed to any device were used as controls. Each plate was retrieved and put in an incubator to count the number of bacterial colonies formed on the plate. There was no significant difference in the number of colonies on plates exposed to devices compared to control, in the number for all devices, or in all bacteria tested in experiments in the 14.4 m3 chamber. These results strongly suggest that these devices have almost no bactericidal effect, at least in space exceeding this volume. Colony formation was suppressed in the glove box in all devices and in all bacteria tested except P. aeruginosa, although the degree of suppression differed among experiments. The colony formation suppression mechanism was analyzed, and indicated that:colony formation did not change even after the removal of Plasmacluster ions, Nanoe particles, or negative ions from the air, while colony formation was decreased drastically by the removal of ozone from space, which was revealed to be generated inevitably during device operation. These results strongly suggest that the

Preparation, Imaging, and Quantification of Bacterial Surface Motility Assays

PubMed Central

Morales-Soto, Nydia; Anyan, Morgen E.; Mattingly, Anne E.; Madukoma, Chinedu S.; Harvey, Cameron W.; Alber, Mark; Déziel, Eric; Kearns, Daniel B.; Shrout, Joshua D.

2015-01-01

Bacterial surface motility, such as swarming, is commonly examined in the laboratory using plate assays that necessitate specific concentrations of agar and sometimes inclusion of specific nutrients in the growth medium. The preparation of such explicit media and surface growth conditions serves to provide the favorable conditions that allow not just bacterial growth but coordinated motility of bacteria over these surfaces within thin liquid films. Reproducibility of swarm plate and other surface motility plate assays can be a major challenge. Especially for more “temperate swarmers” that exhibit motility only within agar ranges of 0.4%-0.8% (wt/vol), minor changes in protocol or laboratory environment can greatly influence swarm assay results. “Wettability”, or water content at the liquid-solid-air interface of these plate assays, is often a key variable to be controlled. An additional challenge in assessing swarming is how to quantify observed differences between any two (or more) experiments. Here we detail a versatile two-phase protocol to prepare and image swarm assays. We include guidelines to circumvent the challenges commonly associated with swarm assay media preparation and quantification of data from these assays. We specifically demonstrate our method using bacteria that express fluorescent or bioluminescent genetic reporters like green fluorescent protein (GFP), luciferase (lux operon), or cellular stains to enable time-lapse optical imaging. We further demonstrate the ability of our method to track competing swarming species in the same experiment. PMID:25938934

Effects of storage temperature on bacterial growth rates and community structure in fresh retail sushi.

PubMed

Hoel, S; Jakobsen, A N; Vadstein, O

2017-09-01

This study was conducted to assess the effects of different storage temperatures (4-20°C), on bacterial concentrations, growth rates and community structure in fresh retail sushi, a popular retail product with a claimed shelf life of 2-3 days. The maximum specific growth rate based on aerobic plate count (APC) at 4°C was 0·06 h -1 and displayed a sixfold increase (0·37 h -1 ) at 20°C. Refrigeration resulted in no growth of hydrogen sulphide (H 2 S)-producing bacteria, but this group had the strongest temperature response. The bacterial community structure was determined by PCR/DGGE (denaturing gradient gel electrophoresis). Multivariate analysis based on Bray-Curtis similarities demonstrated that temperature alone was not the major determinant for the bacterial community structure. The total concentration of aerobic bacteria was the variable that most successfully explained the differences between the communities. The dominating organisms, detected by sequencing of DNA bands excised from the DGGE gel, were Brochothrix thermosphacta and genera of lactic acid bacteria (LAB). The relationship between growth rates and storage temperatures clearly demonstrates that these products are sensitive to deviations from optimal storage temperature, possibly resulting in loss of quality during shelf life. Regardless of the storage temperature, the bacterial communities converged towards a similar structure and density, but the storage temperature determined how fast the community reached its carrying capacity. Little information is available on the microbial composition of ready-to-eat food that are prepared with raw fish, subjected to contamination during handling, and susceptible to microbial growth during cold storage. Moreover, the data are a good first possibility to simulate growth of APC, H 2 S-producing bacteria and LAB under different temperature scenarios that might occur during production, distribution or storage. © 2017 The Society for Applied Microbiology.

Air, water, and surface bacterial contamination in a university-hospital autopsy room.

PubMed

Maujean, Géraldine; Malicier, Daniel; Fanton, Laurent

2012-03-01

Today, little is known about the bacteriological environment of the autopsy room and its potential interest for medico-legal practices. Seven hundred fifty microbiological samples were taken from surface (n = 660), air (n = 48), and water (n = 42) to evaluate it in a French University Forensic Department. Median bacterial counts were compared before and during autopsy for air samples, and before and after autopsy for surface samples, using Wilcoxon matched pairs signed ranks test. Bacterial identification relied on traditional phenotypic methods. Bacterial counts in the air were low before autopsy, increased significantly during procedure, and seemed more linked to the number of people in the room than to an important production of aerosol-containing bacteria. Despite cleaning, human fecal flora was omnipresent on surfaces, which revealed insufficient disinfection. Bacteriological sampling is an easy way to monitor cleaning practices in postmortem rooms, but chiefly a way to improve the reliability of medico-legal proofs of infectious deaths. © 2012 American Academy of Forensic Sciences.

Combination of therapeutic ultrasound with antibiotics interfere with the growth of bacterial culture that colonizes skin ulcers: An in-vitro study.

PubMed

Guirro, Elaine Caldeira de Oliveira; Angelis, Dejanira de Franceschi de; Sousa, Natanael Teixeira Alves de; Guirro, Rinaldo Roberto de Jesus

2016-09-01

Staphylococcus aureus and Escherichia coli are among the major bacterial species that colonize skin ulcers. Therapeutic ultrasound (TUS) produces biophysical effects that are relevant to wound healing; however, its application over a contaminated injury is not evidence-based. The objective of this research was to analyze the effect of TUS on in vitro-isolated S. aureus and E. coli, including the combination of ultrasound and antibiotics, in order to assess their antibiotic action on bacterial susceptibility. For the experiments, the bacterial strains were suspended in saline, then diluted (10(4)CFU/mL) for irradiation (at 1 and 3MHz, 0.5 and 0.8W/cm(2) for 0 and 15min) and the combination treatment of ultrasonication and antibiotics was administered by adding nalidixic acid (S. aureus) and tetracycline (E. coli) at concentrations equivalent to 50% of the minimum inhibitory concentration (MIC). The experiments were carried out in duplicate with six repetitions. The suspensions were inoculated on to Petri plates and incubated at 37°C and the colony forming units (CFUs) were counted after 24h. The results were subjected to the Shapiro-Wilk normality test, followed by parametric ANOVA and Tukey's post hoc test at a significance level of 1%. The results demonstrated that the action of TUS at 1MHz inhibited bacterial growth while at 3MHz, bacterial growth was observed in both species. However, the synergistic combination of ultrasound and antibiotics was able to inhibit the growth of both bacteria completely after 15min of ultrasonication. The results suggest that the action of ultrasound on S. aureus and E. coli are dependent on the oscillation frequency as well as the intensity and time of application. The combination of ultrasound with antibiotics was able to inhibit bacterial growth fully at all frequencies and doses in both species. Copyright © 2016 Elsevier B.V. All rights reserved.

Detection and quantification of intracellular bacterial colonies by automated, high-throughput microscopy.

PubMed

Ernstsen, Christina L; Login, Frédéric H; Jensen, Helene H; Nørregaard, Rikke; Møller-Jensen, Jakob; Nejsum, Lene N

2017-08-01

To target bacterial pathogens that invade and proliferate inside host cells, it is necessary to design intervention strategies directed against bacterial attachment, cellular invasion and intracellular proliferation. We present an automated microscopy-based, fast, high-throughput method for analyzing size and number of intracellular bacterial colonies in infected tissue culture cells. Cells are seeded in 48-well plates and infected with a GFP-expressing bacterial pathogen. Following gentamicin treatment to remove extracellular pathogens, cells are fixed and cell nuclei stained. This is followed by automated microscopy and subsequent semi-automated spot detection to determine the number of intracellular bacterial colonies, their size distribution, and the average number per host cell. Multiple 48-well plates can be processed sequentially and the procedure can be completed in one working day. As a model we quantified intracellular bacterial colonies formed by uropathogenic Escherichia coli (UPEC) during infection of human kidney cells (HKC-8). Urinary tract infections caused by UPEC are among the most common bacterial infectious diseases in humans. UPEC can colonize tissues of the urinary tract and is responsible for acute, chronic, and recurrent infections. In the bladder, UPEC can form intracellular quiescent reservoirs, thought to be responsible for recurrent infections. In the kidney, UPEC can colonize renal epithelial cells and pass to the blood stream, either via epithelial cell disruption or transcellular passage, to cause sepsis. Intracellular colonies are known to be clonal, originating from single invading UPEC. In our experimental setup, we found UPEC CFT073 intracellular bacterial colonies to be heterogeneous in size and present in nearly one third of the HKC-8 cells. This high-throughput experimental format substantially reduces experimental time and enables fast screening of the intracellular bacterial load and cellular distribution of multiple

Intercondylar humerus fracture- parallel plating and its results.

PubMed

Kumar, Sanjiv; Singh, Sudhir; Kumar, Dharmender; Kumar, Neeraj; Verma, Reetu

2015-01-01

Intercondylar fracture of humerus is one of the commonest fractures of young adult and counts for about 30% of all elbow fractures. The treatment of these fractures continues to present challenges despite advances in internal fixation. Although orthogonal plating use to provid adequate functional results in these fractures, parallel plating is said to be mechanically more stable construct thus allowing early mobilization and better range of motion. AIM of the study is to assess the clinical as well functional results of these fractures treated with parallel plating. Prospective study in a tertiary care hospital. A total of 23 fresh patients of intercondylar fracture of humerus from Jan 2013 to May 2014 were included in the study and were treated with parallel plating. These patients were followed at 3, 6, 12, 24 weeks and at 1year of follow up and assessed in terms of time for union, range of motion, MAYO score, DASH score and complication rate. At final follow up Mayo score was 96.32±04.96 from 5.00±01.26 and DASH SCORE was 31.42±2.04 which dropped from 150±05.34, Range of motion improved from 21.38±05.70 to 116.1±07.92 with 100% union rate and complications less than 19%. Parallel plating for intercondylar fracture of humerus is excellent method of fixation and results are similar to those treated with orthogonal plating.

Identifying fecal pollution sources using 3M(™) Petrifilm (™) count plates and antibiotic resistance analysis in the Horse Creek Watershed in Aiken County, SC (USA).

PubMed

Harmon, S Michele; West, Ryan T; Yates, James R

2014-12-01

Sources of fecal coliform pollution in a small South Carolina (USA) watershed were identified using inexpensive methods and commonly available equipment. Samples from the upper reaches of the watershed were analyzed with 3M(™) Petrifilm(™) count plates. We were able to narrow down the study's focus to one particular tributary, Sand River, that was the major contributor of the coliform pollution (both fecal and total) to a downstream reservoir that is heavily used for recreation purposes. Concentrations of total coliforms ranged from 2,400 to 120,333 cfu/100 mL, with sharp increases in coliform counts observed in samples taken after rain events. Positive correlations between turbidity and fecal coliform counts suggested a relationship between fecal pollution and stormwater runoff. Antibiotic resistance analysis (ARA) compared antibiotic resistance profiles of fecal coliform isolates from the stream to those of a watershed-specific fecal source library (equine, waterfowl, canines, and untreated sewage). Known fecal source isolates and unknown isolates from the stream were exposed to six antibiotics at three concentrations each. Discriminant analysis grouped known isolates with an overall average rate of correct classification (ARCC) of 84.3 %. A total of 401 isolates from the first stream location were classified as equine (45.9 %), sewage (39.4 %), waterfowl (6.2 %), and feline (8.5 %). A similar pattern was observed at the second sampling location, with 42.6 % equine, 45.2 % sewage, 2.8 % waterfowl, 0.6 % canine, and 8.8 % feline. While there were slight weather-dependent differences, the vast majority of the coliform pollution in this stream appeared to be from two sources, equine and sewage. This information will contribute to better land use decisions and further justify implementation of low-impact development practices within this urban watershed.

Vulnerability of Bacillus spores and of related genera to physical impaction injury with particular reference to spread-plating.

PubMed

Thomas, P; Sekhar, A C; Mujawar, M M

2014-11-01

To examine whether bacterial spores are vulnerable to impaction injury during standard spread-plating or to other modes of physical impaction. Employing heat-challenged spores of Bacillus pumilus, Bacillus subtilis, Bacillus thuringiensis, Lysinibacillus, Paenibacillus and Brevibacillus spp. from day-4 to day-10 nutrient agar (NA) plates in 50% ethanol, plating the spore suspension to the extent of just drying the agar surface on fresh NA (50-60 s; SP-B) was tested in comparison with the spreader-independent approach of spotting-and-tilt-spreading (SATS), or a brief plating (<10 s; SP-A). Spore CFU was significantly reduced with SP-B in different organisms (23-40%) over SATS independent of the spore size. Comparing 4-, 7- and 10-day-old B. pumilus spores, the former two displayed significant CFU reduction in SP-B indicating a spore age-related effect. Continuous plating for 2-5 min showed a reduction in spore CFU in all organisms depending on plating duration. CFU reduction effect with SP-B was less manifest on refrigerated plates where no friction was experienced but acute on prewarmed and surface-dried plates. Spreader movement over agar surface subsequent to the exhaustion of free moisture proved highly detrimental to spores. A simulated plating study by plating the spores over a plastic film till drying showed a significant reduction in spore CFU. DAPI staining and glass bead-vortexing studies confirmed spore disruption through physical impaction. Bacterial spores are vulnerable to injury during spread-plating or with other forms of physical impaction with variable effects on different genotypes independent of the spore size but altered by spore age. Implications during spore CFU estimations employing spread-plating and during spore surveillance, and the recommendation of SATS as an easier and safer alternative for spore CFU enumeration. © 2014 The Society for Applied Microbiology.

Reflected scatterometry for noninvasive interrogation of bacterial colonies

NASA Astrophysics Data System (ADS)

Kim, Huisung; Doh, Iyll-Joon; Sturgis, Jennifer; Bhunia, Arun K.; Robinson, J. Paul; Bae, Euiwon

2016-10-01

A phenotyping of bacterial colonies on agar plates using forward-scattering diffraction-pattern analysis provided promising classification of several different bacteria such as Salmonella, Vibrio, Listeria, and E. coli. Since the technique is based on forward-scattering phenomena, light transmittance of both the colony and the medium is critical to ensure quality data. However, numerous microorganisms and their growth media allow only limited light penetration and render the forward-scattering measurement a challenging task. For example, yeast, Lactobacillus, mold, and several soil bacteria form colorful and dense colonies that obstruct most of the incoming light passing through them. Moreover, blood agar, which is widely utilized in the clinical field, completely blocks the incident coherent light source used in forward scatterometry. We present a newly designed reflection scatterometer and validation of the resolving power of the instrument. The reflectance-type instrument can acquire backward elastic scatter patterns for both highly opaque media and colonies and has been tested with three different bacterial genera grown on blood agar plates. Cross-validation results show a classification rate above 90% for four genera.

The effect of different precooling rates and cold storage on milk microbiological quality and composition.

PubMed

Paludetti, Lizandra F; Kelly, Alan L; O'Brien, Bernadette; Jordan, Kieran; Gleeson, David

2018-03-01

The objective of this study was to measure the effect of different milk cooling rates, before entering the bulk tank, on the microbiological load and composition of the milk, as well as on energy usage. Three milk precooling treatments were applied before milk entered 3 identical bulk milk tanks: no plate cooler (NP), single-stage plate cooler (SP), and double-stage plate cooler (DP). These precooling treatments cooled the milk to 32.0 ± 1.4°C, 17.0 ± 2.8°C, and 6.0 ± 1.1°C, respectively. Milk was added to the bulk tank twice daily for 72 h, and the tank refrigeration temperature was set at 3°C. The blend temperature within each bulk tank was reduced after each milking event as the volume of milk at 3°C increased simultaneously. The bacterial counts of the milk volumes precooled at different rates did not differ significantly at 0 h of storage or at 24-h intervals thereafter. After 72 h of storage, the total bacterial count of the NP milk was 3.90 ± 0.09 log 10 cfu/mL, whereas that of the precooled milk volumes were 3.77 ± 0.09 (SP) and 3.71 ± 0.09 (DP) log 10 cfu/mL. The constant storage temperature (3°C) over 72 h helped to reduce bacterial growth rates in milk; consequently, milk composition was not affected and minimal, if any, proteolysis occurred. The DP treatment had the highest energy consumption (17.6 ± 0.5 Wh/L), followed by the NP (16.8 ± 2.7 Wh/L) and SP (10.6 ± 1.3 Wh/L) treatments. This study suggests that bacterial count and composition of milk are minimally affected when milk is stored at 3°C for 72 h, regardless of whether the milk is precooled; however, milk entering the tank should have good initial microbiological quality. Considering the numerical differences between bacterial counts, however, the use of the SP or DP precooling systems is recommended to maintain low levels of bacterial counts and reduce energy consumption. The Authors. Published by FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association

A direct plating method for estimating populations of Escherichia coli O157 in bovine manure and manure-based materials.

PubMed

Berry, Elaine D; Wells, James E

2008-11-01

Escherichia coli O157:H7 outbreaks associated with produce consumption have brought attention to livestock manures and manure-based soil amendments as potential sources of pathogens for the contamination of these crops. Procedures for enumeration of E. coli O157:H7 are needed to assess the risks of transmission from these manures and their by-products. A direct plating method employing spiral plating onto CHROMagar O157 was investigated for enumeration of E. coli O157:H7 in feedlot surface material, aged bovine manure, bovine manure compost, and manure-amended soil. In studies utilizing samples spiked with a five-strain cocktail of E. coli O157:H7 at levels ranging from 102 to 10(5) CFU/g of sample, there were strong correlations between the observed and predicted levels of this pathogen. Although the addition of 2.5 mg/liter potassium tellurite and 5 mg/liter novobiocin made the medium more restrictive, these amendments enhanced the ability to identify and enumerate E. coli O157:H7 in feedlot surface material, which contained a higher proportion of fresh feces than did the other three sample types and therefore higher levels of interfering bacterial microflora. The spiral plating method was further assessed to determine its ability to enumerate E. coli O157:H7 in naturally contaminated feedlot surface material. Comparison of E. coli O157:H7 counts in feedlot surface material obtained by the spiral plating method and a most probable number technique were well correlated. We conclude that direct spiral plating onto CHROMagar O157 is effective for estimating E. coli O157:H7 levels in a variety of manures and manure-containing sample types to a lower detection limit of 200 CFU/g. The method has application for determining E. coli O157:H7 concentrations in manures and composts before their sale and use as soil amendments and for measuring the effectiveness of manure treatment processes to reduce or inactivate this pathogen.

Bacterial carriage on the fingernails of OR nurses.

PubMed

Wynd, C A; Samstag, D E; Lapp, A M

1994-11-01

This study provides statistically significant data that demonstrate that chipped fingernail polish or fingernail polish worn longer than four days fosters increased numbers of bacteria on the fingernails of OR nurses after surgical hand scrubs. There were no significant correlations between fingernail length and the numbers of bacterial colonies on the fingernails of the study groups tested after performing a standard surgical hand scrub. A convenience sample of 102 perioperative nurses with either freshly polished fingernails; chipped fingernail polish; or natural, polish-free fingernails participated. The data suggest OR nurses can wear fresh fingernail polish on healthy fingernails without risking increased bacterial counts.

Bacterial dispersion in relation to operating room clothing.

PubMed Central

Whyte, W.; Vesley, D.; Hodgson, R.

1976-01-01

The effect of operating clothing on the dispersal of bacterial particles from the wearers was studied in a dispersal chamber. A comparison was made of six gowns as well as four types of trousers. The gowns were of three basic types, namely a conventional cotton type, disposable types made of non-woven fabric and those of the total-body exhaust system (Charnley type). The dispersal chamber could simulate conditions as expected both in down-flow unidirectional ultra-clean systems and in a conventional turbulent plenum-ventilated system. It was found that the disposable gowns would reduce the dispersal rate by about 30% in the simulated conventionally ventilated system and about 65% in the laminar flow system. The total-body exhaust system (Charnley) would reduce the count by 10-fold in the conventional ventilated system and by 66-fold in the laminar-flow system. The poor performance of the gowns in conventionally ventilated systems was caused by the dispersal of bacterial particles from underneath the gown (about 80%). This was not reduced by the disposable gown and only partially by the Charnley type. This small drop would be further decreased in a conventionally ventilated operating-room as only scrubbed staff would wear the gown. In order to overcome this poor performance in conventionally ventilated operating-rooms impervious trousers would be required. Four types were studied and it was demonstrated that those made either from Ventile or non-woven fabric would reduce the bacterial dispersion fourfold. As these tests had been carried out in an artificial environment checks were carried out in the unidirectional-flow operating-room during total-hip arthroplasty. This was done by comparing conventional cotton gowns with non-woven gowns and total-body exhaust gowns. The results showed good correlation between the operating room and the chamber with the non-woven fabric gown but the total-body exhaust system did not perform as well in the operating room (12-fold

Influence of short distance transportation on tracheal bacterial content and lower airway cytology in horses.

PubMed

Allano, Marion; Labrecque, Olivia; Rodriguez Batista, Edisleidy; Beauchamp, Guy; Bédard, Christian; Lavoie, Jean-Pierre; Leclere, Mathilde

2016-08-01

The aim of this study was to determine the effects of short distance transportation on airway mucus, cytology and bacterial culture to identify potential biases in the diagnosis of airway diseases in referral centres. Eight healthy adult horses were studied using a prospective cross-over design. Mucus scores, tracheal wash (cytology, bacterial culture) and bronchoalveolar lavage fluid (BALF; cytology) were obtained while stabled and following 2.5 h transportation (with and without hay). Neutrophil counts, percentages and BALF neutrophilia frequency increased following transport without hay (P <0.05). No effect was observed on tracheal cytology and bacterial count (P > 0.05). BALF neutrophilia could develop solely as a result of transportation or due to interactions between repeated transports, ambient temperature, head position or other environmental factors. Copyright © 2016 Elsevier Ltd. All rights reserved.

Killing rate of colony count by hydrodynamic cavitation due to square multi-orifice plates

NASA Astrophysics Data System (ADS)

Dong, Zhiyong; Zhao, Wenqian

2018-02-01

Currently,in water supply engineering, the conventional technique of disinfection by chlorination is employed to kill pathogenic microorganisms in raw water. However, chlorine reacts with organic compounds in water and generates disinfection byproducts (DBPs), such as trihalomethanes (THMs), haloacetic acids (HAAs) etc. These byproducts are of carcinogenic, teratogenic and mutagenic effects, which seriously threaten human health. Hydrodynamic cavitation is a novel technique of drinking water disinfection without DBPs. Effects of orifice size, orifice number and orifice layout of multi-orifice plate, cavitation number, cavitation time and orifice velocity on killing pathogenic microorganisms by cavitation were investigated experimentally in a self-developed square multi-orifice plate-type hydrodynamic cavitation device. The experimental results showed that cavitation effects increased with decrease in orifice size and increase in orifice number, cavitation time and orifice velocity. Along with lowering in cavitation number, there was an increase in Reynolds shear stress,thus enhancing the killing rate of pathogenic microorganism in raw water. In addition, the killing rate by staggered orifice layout was greater than that by checkerboard-type orifice layout.

Residual Agar Determination in Bacterial Spores by Electrospray Ionization Mass Spectrometry

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

Wahl, Karen L.; Colburn, Heather A.; Wunschel, David S.

2010-02-15

Presented here is an analytical method to detect residual agar from a bacterial spore sample as an indication of culturing on an agar plate. This method is based on the resolubilization of agar polysaccharide from a bacterial spore sample, enzymatic digestion, followed by electrospray ionization tandem mass spectrometry (ESI-MSn) analysis for detection of a specific agar fragment ion. A range of Bacillus species and strains were selected to demonstrate the effectiveness of this approach. The characteristic agar fragment ion was detected in the spores grown on agar that were washed from 1 to 5 times, irradiated or non-irradiated and notmore » in the spores grown in broth. A sample containing approximately 108 spores is currently needed for confident detection of residual agar from culture on agar plates in the presence of bacterial spores with a limit of detection of approximately 1 ppm agar spiked into a broth-grown spore sample. The results of a proficiency test with 42 blinded samples are presented demonstrating the utility of this method with no false positives and only 3 false negatives for samples that were below the detection level of the method as documented.« less

Effectiveness of a steam cleaning unit for disinfection in a veterinary hospital.

PubMed

Wood, Cheryl L; Tanner, Benjamin D; Higgins, Laura A; Dennis, Jeffrey S; Luempert, Louis G

2014-12-01

To evaluate whether the application of steam to a variety of surface types in a veterinary hospital would effectively reduce the number of bacteria. 5 surface types. Steam was applied as a surface treatment for disinfection to 18 test sites of 5 surface types in a veterinary hospital. A pretreatment sample was obtained by collection of a swab specimen from the left side of each defined test surface. Steam disinfection was performed on the right side of each test surface, and a posttreatment sample was then collected in the same manner from the treated (right) side of each test surface. Total bacteria for pretreatment and posttreatment samples were quantified by heterotrophic plate counts and for Staphylococcus aureus, Pseudomonas spp, and total coliforms by counts on selective media. Significant reductions were observed in heterotrophic plate counts after steam application to dog runs and dog kennel floors. A significant reduction in counts of Pseudomonas spp was observed after steam application to tub sinks. Bacterial counts were reduced, but not significantly, on most other test surfaces that had adequate pretreatment counts for quantification. Development of health-care-associated infections is of increasing concern in human and veterinary medicine. The application of steam significantly reduced bacterial numbers on a variety of surfaces within a veterinary facility. Steam disinfection may prove to be an alternative or adjunct to chemical disinfection within veterinary practices.

Optimizing the position resolution of a Z-stack microchannel plate resistive anode detector for low intensity signals

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

Wiggins, B. B.; Richardson, E.; Siwal, D.

A method for achieving good position resolution of low-intensity electron signals using a microchannel plate resistive anode detector is demonstrated. Electron events at a rate of 7 counts s{sup −1} are detected using a Z-stack microchannel plate. The dependence of position resolution on both the distance and the potential difference between the microchannel plate and resistive anode is investigated. Using standard commercial electronics, a measured position resolution of 170 μm (FWHM) is obtained, which corresponds to an intrinsic resolution of 157 μm (FWHM)

Cellular and soluble components decrease the viable pathogen counts in milk from dairy cows with subclinical mastitis.

PubMed

Koshiishi, Tomoko; Watanabe, Masako; Miyake, Hajime; Hisaeda, Keiichi; Isobe, Naoki

2017-08-10

The present study was undertaken to clarify the factors that reduce the viable pathogen count in milk collected from the udders of subclinical mastitic cows during preservation. Milk was centrifuged to divide somatic cells (cellular components, precipitates) and antimicrobial peptides (soluble components, supernatants without fat layer); each fraction was cultured with bacteria, and the number of viable bacteria was assessed prior to and after culture. In 28.8% of milk samples, we noted no viable bacteria immediately after collection; this value increased significantly after a 5-hr incubation of milk with cellular components but not with soluble components (48.1 and 28.8%, respectively). After culture with cellular components, the numbers of bacteria (excluding Staphylococcus aureus and Streptococcus uberis) and yeast decreased dramatically, although the differences were not statistically significant. After cultivation with soluble components, only yeasts showed a tendency toward decreased mean viability, whereas the mean bacterial counts of S. uberis and T. pyogenes tended to increase after 5-hr preservation with soluble components. These results suggest that most pathogens in high somatic cell count (SCC) milk decreased during preservation at 15 to 25°C, due to both the cellular components and antimicrobial components in the milk. Particularly, the cellular components more potently reduced bacterial counts during preservation.

Bacterial vaginosis: Etiology and modalities of treatment—A brief note

PubMed Central

Kumar, Nikhil; Behera, Beauty; Sagiri, Sai S.; Pal, Kunal; Ray, Sirsendu S.; Roy, Saroj

2011-01-01

A large women population of the world is suffering from a vaginal infection commonly known as bacterial vaginosis. The disease is associated with the decrease in the lactobacilli count in the vagina. Till date, there is a lack of full proof treatment modalities for the cure of the disease. The treatment includes the use of antimicrobials and/or acidifying agents and probiotics, either separately or in combination. This note discusses about the etiology and the various present-day modalities of treatment of bacterial vaginosis. PMID:22219582

Automated food microbiology: potential for the hydrophobic grid-membrane filter.

PubMed Central

Sharpe, A N; Diotte, M P; Dudas, I; Michaud, G L

1978-01-01

Bacterial counts obtained on hydrophobic grid-membrane filters were comparable to conventional plate counts for Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus in homogenates from a range of foods. The wide numerical operating range of the hydrophobic grid-membrane filters allowed sequential diluting to be reduced or even eliminated, making them attractive as components in automated systems of analysis. Food debris could be rinsed completely from the unincubated hydrophobic grid-membrane filter surface without affecting the subsequent count, thus eliminating the possibility of counting food particles, a common source of error in electronic counting systems. PMID:100054

Diagnostic markers of serious bacterial infections in febrile infants younger than 90 days old.

PubMed

Nosrati, Adi; Ben Tov, Amir; Reif, Shimon

2014-02-01

The aim of this study was to assess correlations between demographic, clinical and laboratory characteristics and the risk of serious bacterial infection (SBI) in febrile <90-day-old infants. Medical records of all infants younger than 90 days old hospitalized at Dana-Dwek Children's Hospital (2006-2008) for evaluation of fever were retrospectively reviewed. Data on clinical, laboratory and demographic characteristics were retrieved and evaluated. Forty-eight of the 401 study infants (12%) had SBI: most of them had urinary tract infection (43 infants; 90% of all SBI), three infants had bacteremia, one had bacterial pneumonia and one had bacterial meningitis. Significant independent clinical predictors for the diagnosis of SBI included duration of fever, absence of rhinitis and the absence of lung and skin manifestations. Significant independent laboratory predictors were absolute neutrophil count (ANC), platelets, blood urea nitrogen and C-reactive protein (CRP) level. On receiver operating characteristic curve analysis, the CRP area under the curve (0.819) was significantly superior to ANC and leukocyte count. Of the clinical and laboratory variables selected for evaluation, qualitative CRP was the strongest independent predictor for diagnosing SBI and a significantly better diagnostic marker than clinical characteristics, ANC and white blood cell count. © 2013 The Authors. Pediatrics International © 2013 Japan Pediatric Society.

Improvements in Boron Plate Coating Technology for Higher Efficiency Neutron Detection and Coincidence Counting Error Reduction

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

Menlove, Howard Olsen; Henzlova, Daniela

This informal report presents the measurement data and information to document the performance of the advanced Precision Data Technology, Inc. (PDT) sealed cell boron-10 plate neutron detector that makes use of the advanced coating materials and procedures. In 2015, PDT changed the boron coating materials and application procedures to significantly increase the efficiency of their basic corrugated plate detector performance. A prototype sealed cell unit was supplied to LANL for testing and comparison with prior detector cells. Also, LANL had reference detector slabs from the original neutron collar (UNCL) and the new Antech UNCL with the removable 3He tubes. Themore » comparison data is presented in this report.« less

Direct quantification of bacterial biomass in influent, effluent and activated sludge of wastewater treatment plants by using flow cytometry.

PubMed

Foladori, P; Bruni, L; Tamburini, S; Ziglio, G

2010-07-01

A rapid multi-step procedure, potentially amenable to automation, was proposed for quantifying viable and active bacterial cells, estimating their biovolume using flow cytometry (FCM) and to calculate their biomass within the main stages of a wastewater treatment plant: raw wastewater, settled wastewater, activated sludge and effluent. Fluorescent staining of bacteria using SYBR-Green I + Propidium Iodide (to discriminate cell integrity or permeabilisation) and BCECF-AM (to identify enzymatic activity) was applied to count bacterial cells by FCM. A recently developed specific procedure was applied to convert Forward Angle Light Scatter measured by FCM into the corresponding bacterial biovolume. This conversion permits the calculation of the viable and active bacterial biomass in wastewater, activated sludge and effluent, expressed as Volatile Suspended Solids (VSS) or particulate Chemical Oxygen Demand (COD). Viable bacterial biomass represented only a small part of particulate COD in raw wastewater (4.8 +/- 2.4%), settled wastewater (10.7 +/- 3.1%), activated sludge (11.1 +/- 2.1%) and effluent (3.2 +/- 2.2%). Active bacterial biomass counted for a percentage of 30-47% of the viable bacterial biomass within the stages of the wastewater treatment plant. Copyright 2010 Elsevier Ltd. All rights reserved.

Bacterial populations associated with the dirty area of a South African poultry abattoir.

PubMed

Geornaras, I; de Jesus, A E; von Holy, A

1998-06-01

Bacterial populations associated with three sample types from the neck region of poultry carcasses in the dirty area of an abattoir were characterized. Sample types before and after scalding were skin only, feathers only, and a skin and feather combination. The neck skin of carcasses after the defeathering processing stage was also sampled. Bacterial populations associated with water from the scald tank, rubber fingers at the exit of the defeathering machine, and air in the dirty area were also characterized. Bacterial colonies (751) were randomly isolated from yeast extract-supplemented tryptone soya agar plates exhibiting 30 to 300 colonies. Micrococcus spp. were isolated in the highest proportion from pre-and postscalded carcass samples (63.5 to 86.1% of isolates), regardless of the sample type. Conversely, Enterobacteriaceae (40.3%), Acinetobacter (19.4%), and Aeromonas/Vibrio (12.5%) species predominated on neck skin samples taken from mechanically defeathered carcasses. Isolates from the rubber fingers were, however, predominantly Micrococcus spp. (94.4%). Bacterial groups isolated in the highest proportion from scald tank water samples were Micrococcus spp. (38.3%), species of Enterobacteriaceae (29.1%), and lactic acid bacteria (17.0%). Corynebacterium spp., species of Enterobacteriaceae, and Micrococcus spp. were dominant on air settle plates.

Designing primers and evaluation of the efficiency of propidium monoazide - Quantitative polymerase chain reaction for counting the viable cells of Lactobacillus gasseri and Lactobacillus salivarius.

PubMed

Lai, Chieh-Hsien; Wu, Sih-Rong; Pang, Jen-Chieh; Ramireddy, Latha; Chiang, Yu-Cheng; Lin, Chien-Ku; Tsen, Hau-Yang

2017-07-01

The purpose of this study is to evaluate the efficiency of using propidium monoazide (PMA) real-time quantitative polymerase chain reaction (qPCR) to count the viable cells of Lactobacillus gasseri and Lactobacillus salivarius in probiotic products. Based on the internal transcription spacer and 23S rRNA genes, two primer sets specific for these two Lactobacillus species were designed. For a probiotic product, the total deMan Rogosa Sharpe plate count was 8.65±0.69 log CFU/g, while for qPCR, the cell counts of L. gasseri and L. salivarius were 8.39±0.14 log CFU/g and 8.57±0.24 log CFU/g, respectively. Under the same conditions, for its heat-killed product, qPCR counts for L. gasseri and L. salivarius were 6.70±0.16 log cells/g and 7.67±0.20 log cells/g, while PMA-qPCR counts were 5.33±0.18 log cells/g and 5.05±0.23 log cells/g, respectively. For cell dilutions with a viable cell count of 8.5 log CFU/mL for L. gasseri and L. salivarius, after heat killing, the PMA-qPCR count for both Lactobacillus species was near 5.5 log cells/mL. When the PMA-qPCR counts of these cell dilutions were compared before and after heat killing, although some DNA might be lost during the heat killing, significant qPCR signals from dead cells, i.e., about 4-5 log cells/mL, could not be reduced by PMA treatment. Increasing PMA concentrations from 100 μM to 200 μM or light exposure time from 5 minutes to 15 minutes had no or, if any, only minor effect on the reduction of qPCR signals from their dead cells. Thus, to differentiate viable lactic acid bacterial cells from dead cells using the PMA-qPCR method, the efficiency of PMA to reduce the qPCR signals from dead cells should be notable. Copyright © 2016. Published by Elsevier B.V.

Effect of Punica granatum L. Flower Water Extract on Five Common Oral Bacteria and Bacterial Biofilm Formation on Orthodontic Wire

PubMed Central

VAHID DASTJERDI, Elahe; ABDOLAZIMI, Zahra; GHAZANFARIAN, Marzieh; AMDJADI, Parisa; KAMALINEJAD, Mohammad; MAHBOUBI, Arash

2014-01-01

Background: Use of herbal extracts and essences as natural antibacterial compounds has become increasingly popular for the control of oral infectious diseases. Therefore, finding natural antimicrobial products with the lowest side effects seems necessary. The present study sought to assess the effect of Punica granatum L. water extract on five oral bacteria and bacterial biofilm formation on orthodontic wire. Methods: Antibacterial property of P. granatum L. water extract was primarily evaluated in brain heart infusion agar medium using well-plate method. The minimum inhibitory concentration and minimum bactericidal concentration were determined by macro-dilution method. The inhibitory effect on orthodontic wire bacterial biofilm formation was evaluated using viable cell count in biofilm medium. At the final phase, samples were fixed and analyzed by Scanning Electron Microscopy. Results: The growth inhibition zone diameter was proportional to the extract concentration. The water extract demonstrated the maximum antibacterial effect on Streptococcus sanguinis ATCC 10556 with a minimum inhibitory concentration of 6.25 mg/ml and maximum bactericidal effect on S. sanguinis ATCC 10556 and S. sobrinus ATCC 27607 with minimum bactericidal concentration of 25 mg/ml. The water extract decreased bacterial biofilm formation by S. sanguinis, S. sobrinus, S. salivarius, S. mutans ATCC 35608 and E. faecalis CIP 55142 by 93.7–100%, 40.6–99.9%, 85.2–86.5%, 66.4–84.4% and 35.5–56.3% respectively. Conclusion: Punica granatum L. water extract had significant antibacterial properties against 5 oral bacteria and prevented orthodontic wire bacterial biofilm formation. However, further investigations are required to generalize these results to the clinical setting. PMID:26171362

Rapid assessment of viable but non-culturable Bacillus coagulans MTCC 5856 in commercial formulations using Flow cytometry.

PubMed

Majeed, Muhammed; Majeed, Shaheen; Nagabhushanam, Kalyanam; Punnapuzha, Ardra; Philip, Sheena; Mundkur, Lakshmi

2018-01-01

Accurate enumeration of bacterial count in probiotic formulation is imperative to ensure that the product adheres to regulatory standards and citation in consumer product label. Standard methods like plate count, can enumerate only replicating bacterial population under selected culture conditions. Viable but non culturable bacteria (VBNC) retain characteristics of living cells and can regain cultivability by a process known as resuscitation. This is a protective mechanism adapted by bacteria to evade stressful environmental conditions. B. coagulans MTCC 5856(LactoSpore®) is a probiotic endospore which can survive for decades in hostile environments without dividing. In the present study, we explored the use of flow cytometry to enumerate the viable count of B. coagulans MTCC 5856 under acidic and alkaline conditions, high temperature and in commercial formulations like compressed tablets and capsules. Flow cytometry (FCM) was comparable to plate count method when the spores were counted at physiological conditions. We show that VBNC state is induced in B. coagulans MTCC 5856by high temperature and acidic pH. The cells get resuscitated under physiological conditions and FCM was sensitive to detect the VBNC spores. Flow cytometry showed excellent ability to assess the viable spore count in commercial probiotic formulations of B. coagulans MTCC 5856. The results establish Flow cytometry as a reliable method to count viable bacteria in commercial probiotic preparations. Sporulation as well as existence as VBNC could contribute to the extreme stability of B. coagulans MTCC 5856.

Rapid assessment of viable but non-culturable Bacillus coagulans MTCC 5856 in commercial formulations using Flow cytometry

PubMed Central

Majeed, Muhammed; Majeed, Shaheen; Nagabhushanam, Kalyanam; Punnapuzha, Ardra; Philip, Sheena

2018-01-01

Accurate enumeration of bacterial count in probiotic formulation is imperative to ensure that the product adheres to regulatory standards and citation in consumer product label. Standard methods like plate count, can enumerate only replicating bacterial population under selected culture conditions. Viable but non culturable bacteria (VBNC) retain characteristics of living cells and can regain cultivability by a process known as resuscitation. This is a protective mechanism adapted by bacteria to evade stressful environmental conditions. B. coagulans MTCC 5856(LactoSpore®) is a probiotic endospore which can survive for decades in hostile environments without dividing. In the present study, we explored the use of flow cytometry to enumerate the viable count of B. coagulans MTCC 5856 under acidic and alkaline conditions, high temperature and in commercial formulations like compressed tablets and capsules. Flow cytometry (FCM) was comparable to plate count method when the spores were counted at physiological conditions. We show that VBNC state is induced in B. coagulans MTCC 5856by high temperature and acidic pH. The cells get resuscitated under physiological conditions and FCM was sensitive to detect the VBNC spores. Flow cytometry showed excellent ability to assess the viable spore count in commercial probiotic formulations of B. coagulans MTCC 5856. The results establish Flow cytometry as a reliable method to count viable bacteria in commercial probiotic preparations. Sporulation as well as existence as VBNC could contribute to the extreme stability of B. coagulans MTCC 5856. PMID:29474436

A comparison of legionella and other bacteria concentrations in cooling tower water

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

Cappabianca, R.M.; Jurinski, N.B.; Jurinski, J.B.

1994-05-01

A field study was conducted in which water samples collected from air conditioning cooling water reservoirs of high-rise buildings throughout an urban area were assayed for Legionella and for total bacteria. Buildings included within the study had ongoing biocidal treatment programs for the cooling towers. Separate sample analyses were performed to measure the viable colony concentrations of total bacteria and of Legionella in the process waters. The occurrence and viable counts of Legionella in 304 environmental water samples were determined by inoculating them onto plates of buffered charcoal yeast extract (BCYE) agar medium (a presumptive screening method). The samples weremore » collected during summer months between July and September. BCYE plate cultures of 50 (16.4%) of the samples yielded Legionella with viable counts ranging from 2 to 608 colony forming units per milliliter. In the water samples, 281 (92.4%) yielded viable counts of bacteria that ranged from 9 to 1.2 x 10{sup 6} per milliliter. This study demonstrates that Legionella are commonly present in the water of air conditioning cooling towers and that there is no significant correlation between concurrently sampled culture plate counts of Legionella and total bacteria plate counts. Correspondingly, there is no demonstrated validity for use of total bacterial counts as an inferential surrogate for the concentration of Legionella in the water. 19 refs., 3 figs., 1 tab.« less

Cultivation and qPCR Detection of Pathogenic and Antibiotic-Resistant Bacterial Establishment in Naive Broiler Houses.

PubMed

Brooks, J P; McLaughlin, M R; Adeli, A; Miles, D M

2016-05-01

Conventional commercial broiler production involves the rearing of more than 20,000 broilers in a single confined space for approximately 6.5 wk. This environment is known for harboring pathogens and antibiotic-resistant bacteria, but studies have focused on previously established houses with mature litter microbial populations. In the current study, a set of three naive houses were followed from inception through 11 broiler flocks and monitored for ambient climatic conditions, bacterial pathogens, and antibiotic resistance. Within the first 3 wk of the first flock cycle, 100% of litter samples were positive for and , whereas was cultivation negative but PCR positive. Antibiotic resistance genes were ubiquitously distributed throughout the litter within the first flock, approaching 10 to 10 genomic units g. Preflock litter levels were approximately 10 CFU g for heterotrophic plate count bacteria, whereas midflock levels were >10 colony forming units (CFU) g; other indicators demonstrated similar increases. The influence of intrahouse sample location was minor. In all likelihood, given that preflock levels were negative for pathogens and antibiotic resistance genes and 4 to 5 Log lower than flock levels for indicators, incoming birds most likely provided the colonizing microbiome, although other sources were not ruled out. Most bacterial groups experienced a cyclical pattern of litter contamination seen in other studies, whereas microbial stabilization required approximately four flocks. This study represents a first-of-its-kind view into the time required for bacterial pathogens and antibiotic resistance to colonize and establish in naive broiler houses. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

WBC count

MedlinePlus

Leukocyte count; White blood cell count; White blood cell differential; WBC differential; Infection - WBC count; Cancer - WBC count ... called leukopenia. A count less than 4,500 cells per microliter (4.5 × 10 9 /L) is ...

High temperature, short time pasteurization temperatures inversely affect bacterial numbers during refrigerated storage of pasteurized fluid milk.

PubMed

Ranieri, M L; Huck, J R; Sonnen, M; Barbano, D M; Boor, K J

2009-10-01

The grade A Pasteurized Milk Ordinance specifies minimum processing conditions of 72 degrees C for at least 15 s for high temperature, short time (HTST) pasteurized milk products. Currently, many US milk-processing plants exceed these minimum requirements for fluid milk products. To test the effect of pasteurization temperatures on bacterial numbers in HTST pasteurized milk, 2% fat raw milk was heated to 60 degrees C, homogenized, and treated for 25 s at 1 of 4 different temperatures (72.9, 77.2, 79.9, or 85.2 degrees C) and then held at 6 degrees C for 21 d. Aerobic plate counts were monitored in pasteurized milk samples at d 1, 7, 14, and 21 postprocessing. Bacterial numbers in milk processed at 72.9 degrees C were lower than in milk processed at 85.2 degrees C on each sampling day, indicating that HTST fluid milk-processing temperatures significantly affected bacterial numbers in fluid milk. To assess the microbial ecology of the different milk samples during refrigerated storage, a total of 490 psychrotolerant endospore-forming bacteria were identified using DNA sequence-based subtyping methods. Regardless of processing temperature, >85% of the isolates characterized at d 0, 1, and 7 postprocessing were of the genus Bacillus, whereas more than 92% of isolates characterized at d 14 and 21 postprocessing were of the genus Paenibacillus, indicating that the predominant genera present in HTST-processed milk shifted from Bacillus spp. to Paenibacillus spp. during refrigerated storage. In summary, 1) HTST processing temperatures affected bacterial numbers in refrigerated milk, with higher bacterial numbers in milk processed at higher temperatures; 2) no significant association was observed between genus isolated and pasteurization temperature, suggesting that the genera were not differentially affected by the different processing temperatures; and 3) although typically present at low numbers in raw milk, Paenibacillus spp. are capable of growing to numbers that can

Interactions of Seedborne Bacterial Pathogens with Host and Non-Host Plants in Relation to Seed Infestation and Seedling Transmission

PubMed Central

Dutta, Bhabesh; Gitaitis, Ronald; Smith, Samuel; Langston, David

2014-01-01

The ability of seed-borne bacterial pathogens (Acidovorax citrulli, Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato, Xanthomonas euvesicatoria, and Pseudomonas syringae pv. glycinea) to infest seeds of host and non-host plants (watermelon, tomato, pepper, and soybean) and subsequent pathogen transmission to seedlings was investigated. A non-pathogenic, pigmented strain of Serratia marcescens was also included to assess a null-interacting situation with the same plant species. Flowers of host and non-host plants were inoculated with 1×106 colony forming units (CFUs)/flower for each bacterial species and allowed to develop into fruits or umbels (in case of onion). Seeds harvested from each host/non-host bacterial species combination were assayed for respective bacteria by plating on semi-selective media. Additionally, seedlots for each host/non-host bacterial species combination were also assayed for pathogen transmission by seedling grow-out (SGO) assays under greenhouse conditions. The mean percentage of seedlots infested with compatible and incompatible pathogens was 31.7 and 30.9% (by plating), respectively and they were not significantly different (P = 0.67). The percentage of seedlots infested with null-interacting bacterial species was 16.8% (by plating) and it was significantly lower than the infested lots generated with compatible and incompatible bacterial pathogens (P = 0.03). None of the seedlots with incompatible/null-interacting bacteria developed symptoms on seedlings; however, when seedlings were assayed for epiphytic bacterial presence, 19.5 and 9.4% of the lots were positive, respectively. These results indicate that the seeds of non-host plants can become infested with incompatible and null-interacting bacterial species through flower colonization and they can be transmitted via epiphytic colonization of seedlings. In addition, it was also observed that flowers and seeds of non-host plants can be colonized

Characterization of bacterial pathogens in rural and urban irrigation water.

PubMed

Aijuka, Matthew; Charimba, George; Hugo, Celia J; Buys, Elna M

2015-03-01

The study aimed to compare the bacteriological quality of an urban and rural irrigation water source. Bacterial counts, characterization, identification and diversity of aerobic bacteria were determined. Escherichia coli isolated from both sites was subjected to antibiotic susceptibility testing, virulence gene (Stx1/Stx2 and eae) determination and (GTG)5 Rep-PCR fingerprinting. Low mean monthly counts for aerobic spore formers, anaerobic spore formers and Staphylococcus aureus were noted although occasional spikes were observed. The most prevalent bacterial species at both sites were Bacillus spp., E. coli and Enterobacter spp. In addition, E. coli and Bacillus spp. were most prevalent in winter and summer respectively. Resistance to at least one antibiotic was 84% (rural) and 83% (urban). Highest resistance at both sites was to cephalothin and ampicillin. Prevalence of E. coli possessing at least one virulence gene (Stx1/Stx2 and eae) was 15% (rural) and 42% (urban). All (rural) and 80% (urban) of E. coli possessing virulence genes showed antibiotic resistance. Complete genetic relatedness (100%) was shown by 47% of rural and 67% of urban E. coli isolates. Results from this study show that surface irrigation water sources regardless of geographical location and surrounding land-use practices can be reservoirs of similar bacterial pathogens.

Serum bactericidal assay for the evaluation of typhoid vaccine using a semi-automated colony-counting method.

PubMed

Jang, Mi Seon; Sahastrabuddhe, Sushant; Yun, Cheol-Heui; Han, Seung Hyun; Yang, Jae Seung

2016-08-01

Typhoid fever, mainly caused by Salmonella enterica serovar Typhi (S. Typhi), is a life-threatening disease, mostly in developing countries. Enzyme-linked immunosorbent assay (ELISA) is widely used to quantify antibodies against S. Typhi in serum but does not provide information about functional antibody titers. Although the serum bactericidal assay (SBA) using an agar plate is often used to measure functional antibody titers against various bacterial pathogens in clinical specimens, it has rarely been used for typhoid vaccines because it is time-consuming and labor-intensive. In the present study, we established an improved SBA against S. Typhi using a semi-automated colony-counting system with a square agar plate harboring 24 samples. The semi-automated SBA efficiently measured bactericidal titers of sera from individuals immunized with S. Typhi Vi polysaccharide vaccines. The assay specifically responded to S. Typhi Ty2 but not to other irrelevant enteric bacteria including Vibrio cholerae and Shigella flexneri. Baby rabbit complement was more appropriate source for the SBA against S. Typhi than complements from adult rabbit, guinea pig, and human. We also examined the correlation between SBA and ELISA for measuring antibody responses against S. Typhi using pre- and post-vaccination sera from 18 human volunteers. The SBA titer showed a good correlation with anti-Vi IgG quantity in the serum as determined by Spearman correlation coefficient of 0.737 (P < 0.001). Taken together, the semi-automated SBA might be efficient, accurate, sensitive, and specific enough to measure functional antibody titers against S. Typhi in sera from human subjects immunized with typhoid vaccines. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Inflammatory response in mixed viral-bacterial community-acquired pneumonia.

PubMed

Bello, Salvador; Mincholé, Elisa; Fandos, Sergio; Lasierra, Ana B; Ruiz, María A; Simon, Ana L; Panadero, Carolina; Lapresta, Carlos; Menendez, Rosario; Torres, Antoni

2014-07-29

The role of mixed pneumonia (virus+bacteria) in community-acquired pneumonia (CAP) has been described in recent years. However, it is not known whether the systemic inflammatory profile is different compared to monomicrobial CAP. We wanted to investigate this profile of mixed viral-bacterial infection and to compare it to monomicrobial bacterial or viral CAP. We measured baseline serum procalcitonin (PCT), C reactive protein (CRP), and white blood cell (WBC) count in 171 patients with CAP with definite etiology admitted to a tertiary hospital: 59 (34.5%) bacterial, 66 (39.%) viral and 46 (27%) mixed (viral-bacterial). Serum PCT levels were higher in mixed and bacterial CAP compared to viral CAP. CRP levels were higher in mixed CAP compared to the other groups. CRP was independently associated with mixed CAP. CRP levels below 26 mg/dL were indicative of an etiology other than mixed in 83% of cases, but the positive predictive value was 45%. PCT levels over 2.10 ng/mL had a positive predictive value for bacterial-involved CAP versus viral CAP of 78%, but the negative predictive value was 48%. Mixed CAP has a different inflammatory pattern compared to bacterial or viral CAP. High CRP levels may be useful for clinicians to suspect mixed CAP.

Differentiation of bacterial colonies and temporal growth patterns using hyperspectral imaging

NASA Astrophysics Data System (ADS)

Mehrübeoglu, Mehrube; Buck, Gregory W.; Livingston, Daniel W.

2014-09-01

Detection and identification of bacteria are important for health and safety. Hyperspectral imaging offers the potential to capture unique spectral patterns and spatial information from bacteria which can then be used to detect and differentiate bacterial species. Here, hyperspectral imaging has been used to characterize different bacterial colonies and investigate their growth over time. Six bacterial species (Pseudomonas fluorescens, Escherichia coli, Serratia marcescens, Salmonella enterica, Staphylococcus aureus, Enterobacter aerogenes) were grown on tryptic soy agar plates. Hyperspectral data were acquired immediately after, 24 hours after, and 96 hours after incubation. Spectral signatures from bacterial colonies demonstrated repeatable measurements for five out of six species. Spatial variations as well as changes in spectral signatures were observed across temporal measurements within and among species at multiple wavelengths due to strengthening or weakening reflectance signals from growing bacterial colonies based on their pigmentation. Between-class differences and within-class similarities were the most prominent in hyperspectral data collected 96 hours after incubation.

Plating Bacteriophage M13.

PubMed

Green, Michael R; Sambrook, Joseph

2017-10-03

A plaque of bacteriophage M13 derives from infection of a single bacterium by a single virus particle. The progeny particles infect neighboring bacteria, which, in turn, release another generation of daughter virus particles. If the bacteria are growing in semisolid medium (e.g., containing agar or agarose), then the diffusion of the progeny particles is limited. Cells infected with bacteriophage M13 are not killed, but have a longer generation time than uninfected Escherichia coli In consequence, plaques appear as areas of slower-growing cells on a faster-growing lawn of bacterial cells. This protocol describes plating of bacteriophage M13 stocks. Plaques are readily detectable on top agar after 4-8 h of incubation at 37°C. © 2017 Cold Spring Harbor Laboratory Press.

MALDI-TOF mass spectrometry and high-consequence bacteria: safety and stability of biothreat bacterial sample testing in clinical diagnostic laboratories.

PubMed

Tracz, Dobryan M; Tober, Ashley D; Antonation, Kym S; Corbett, Cindi R

2018-03-01

We considered the application of MALDI-TOF mass spectrometry for BSL-3 bacterial diagnostics, with a focus on the biosafety of live-culture direct-colony testing and the stability of stored extracts. Biosafety level 2 (BSL-2) bacterial species were used as surrogates for BSL-3 high-consequence pathogens in all live-culture MALDI-TOF experiments. Viable BSL-2 bacteria were isolated from MALDI-TOF mass spectrometry target plates after 'direct-colony' and 'on-plate' extraction testing, suggesting that the matrix chemicals alone cannot be considered sufficient to inactivate bacterial culture and spores in all samples. Sampling of the instrument interior after direct-colony analysis did not recover viable organisms, suggesting that any potential risks to the laboratory technician are associated with preparation of the MALDI-TOF target plate before or after testing. Secondly, a long-term stability study (3 years) of stored MALDI-TOF extracts showed that match scores can decrease below the threshold for reliable species identification (<1.7), which has implications for proficiency test panel item storage and distribution.

Evaluation of the Minnesota Easy Culture System II Bi-Plate and Tri-Plate for identification of common mastitis pathogens in milk.

PubMed

Royster, E; Godden, S; Goulart, D; Dahlke, A; Rapnicki, P; Timmerman, J

2014-01-01

The objective of this study was to validate use of the Minnesota Easy Culture System II Bi-Plate and Tri-Plate (University of Minnesota Laboratory for Udder Health, St. Paul) to identify common mastitis pathogens in milk. A total of 283 quarter and composite milk samples submitted to the University of Minnesota Laboratory for Udder Health during the spring of 2010 were cultured simultaneously using 3 methods: standard laboratory culture (reference method) and the Minnesota Easy Culture System II Bi-Plate and Tri-Plate methods. Bi-Plate and Tri-Plate cultures were incubated for 18 to 24h and interpreted by 2 independent, untrained readers within 5h of each other. An experienced technician completed the standard laboratory culture. For each sample, all 3 study personnel recorded the culture result (yes/no) for each of the following diagnostic categories: no bacterial growth (NG), mixed (2 organisms), contaminated (3 or more organisms), gram-positive (GP), gram-negative (GN), Staphylococcus spp., Streptococcus spp., Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Enterococcus spp., Staphylococcus aureus, coagulase-negative staphylococci, Escherichia coli, Klebsiella spp., and other. For each category, the prevalence, sensitivity, specificity, accuracy, and predictive values of a positive and negative test were calculated, and the agreement between readers and between each reader and the laboratory was assessed. Specificity, overall accuracy, and negative predictive values were generally high (>80%) for the Bi-Plate and Tri-Plate for each category. Sensitivity and positive predictive values were intermediate (>60%) or high (>80%) for the broad categories of NG, GP, GN, Staphylococcus spp. and Streptococcus spp., and for Staph. aureus, but were generally lower (<60%) for other more specific categories. Similarly, interreader agreement (kappa value) was moderate to substantial (40-80%) for the broad categories of NG, GP, GN, Staphylococcus spp

The impact of shrimp farming effluent on bacterial communities in mangrove waters, Ceará, Brazil.

PubMed

Sousa, O V; Macrae, A; Menezes, F G R; Gomes, N C M; Vieira, R H S F; Mendonça-Hagler, L C S

2006-12-01

The effects of shrimp farm effluents on bacterial communities in mangroves have been infrequently reported. Classic and molecular biology methods were used to survey bacterial communities from four mangroves systems. Water temperature, salinity, pH, total heterotrophic bacteria and maximum probable numbers of Vibrio spp. were investigated. Genetic profiles of bacterial communities were also characterized by polymerase chain reaction (PCR) amplification of eubacterial and Vibrio 16S rDNA using denaturing gradient gel electrophoresis (DGGE). Highest heterotrophic counts were registered in the mangrove not directly polluted by shrimp farming. The Enterobacteriaceae and Chryseomonas luteola dominated the heterotrophic isolates. Vibrio spp. pathogenic to humans and shrimps were identified. Eubacterial genetic profiles suggest a shared community structure independent of mangrove system. Vibrio genetic profiles were mangrove specific. Neither microbial counts nor genetic profiling revealed a significant decrease in species richness associated with shrimp farm effluent. The complex nature of mangrove ecosystems and their microbial communities is discussed.

Inoculation of a phenanthrene-degrading endophytic bacterium reduces the phenanthrene level and alters the bacterial community structure in wheat.

PubMed

Liu, Juan; Xiang, Yanbing; Zhang, Zhiming; Ling, Wanting; Gao, Yanzheng

2017-06-01

Colonization by polycyclic aromatic hydrocarbon (PAH)-degrading endophytic bacteria (PAHDEB) can reduce the PAH contamination risk in plant. However, little information is available on the impact of PAHDEB colonization on the endophytic bacterial community of inner plant tissues. A phenanthrene-degrading endophytic bacterium (PDEB), Massilia sp. Pn2, was inoculated onto the roots of wheat and subjected to greenhouse container experiments. The endophytic bacterial community structure in wheat was investigated using high-throughput sequencing technology. The majority of endophytic bacteria in wheat were Proteobacteria, and the dominant genus was Pseudomonas. Phenanthrene contamination clearly increased the diversity of endophytic bacteria in wheat. The cultivable endophytic bacteria counts in wheat decreased with increasing the level of phenanthrene contamination; the endophytic bacterial community structure changed correspondingly, and the bacterial richness first increased and then decreased. Inoculation of strain Pn2 reduced the phenanthrene contamination in wheat, enlarged the biomass of wheat roots, changed the bacterial community structure and enhanced the cell counts, diversity and richness of endophytic bacteria in phenanthrene-contaminated wheat in a contamination level-dependent manner. The findings of this investigation provide insight into the responses of endophytic bacterial community in plant to external PAH contamination and PAHDEB colonization.

Interactions of plaunotol with bacterial membranes.

PubMed

Koga, T; Watanabe, H; Kawada, H; Takahashi, K; Utsui, Y; Domon, H; Ishii, C; Narita, T; Yasuda, H

1998-08-01

Plaunotol, a cytoprotective antiulcer agent, has a bactericidal effect against Helicobacter pylori, which may result from interaction of this compound with the bacterial cell membrane. The purpose of the present study was to confirm that plaunotol interacts with the H. pylori membrane. Membrane fluidities were measured using two stearic acid spin labels, namely 5-doxyl-stearic acid (in which the nitroxide group is located in the upper portion of the bacterial cell membrane) and 16-doxyl-stearic acid methyl ester (in which the nitroxide group is located deeper in the bacterial cell membrane), by means of electron spin resonance. The membrane fluidities of plaunotol-treated cells were significantly increased in the measurements made using the two spin labels. We also attempted to isolate plaunotol-resistant H. pylori in vitro by two different methods. To assess the level of resistance that could be reached, H. pylori was passaged five times on an agar plate containing subinhibitory concentrations of plaunotol or metronidazole. To measure the rate of development of resistance, H. pylori was grown with subinhibitory concentrations (0.25 x MIC) of plaunotol or metronidazole, and quantitatively plated on to medium containing 4 x MIC of the compounds. This treatment was repeated once more. No plaunotol-resistant colonies were selected by the two methods. H. pylori developed resistance to metronidazole easily and at a relatively high rate. The mechanism by which plaunotol directly fluidizes and destroys the H. pylori membrane might make it difficult for this organism to develop resistance to plaunotol. It was confirmed that the bactericidal effects of plaunotol were also shown against Staphylococcus aureus, Streptococcus pneumoniae, Neisseria gonorrhoeae, Moraxella catarrhalis and Haemophilus influenzae. No such effect was seen against Escherichia coli and Pseudomonas aeruginosa.

Hydrodynamics of bacterial colonies: A model

NASA Astrophysics Data System (ADS)

Lega, J.; Passot, T.

2003-03-01

We propose a hydrodynamic model for the evolution of bacterial colonies growing on soft agar plates. This model consists of reaction-diffusion equations for the concentrations of nutrients, water, and bacteria, coupled to a single hydrodynamic equation for the velocity field of the bacteria-water mixture. It captures the dynamics inside the colony as well as on its boundary and allows us to identify a mechanism for collective motion towards fresh nutrients, which, in its modeling aspects, is similar to classical chemotaxis. As shown in numerical simulations, our model reproduces both usual colony shapes and typical hydrodynamic motions, such as the whirls and jets recently observed in wet colonies of Bacillus subtilis. The approach presented here could be extended to different experimental situations and provides a general framework for the use of advection-reaction-diffusion equations in modeling bacterial colonies.

Surface Bacterial-Spore Assay Using Tb3+/DPA Luminescence

NASA Technical Reports Server (NTRS)

Ponce, Adrian

2007-01-01

Equipment and a method for rapidly assaying solid surfaces for contamination by bacterial spores are undergoing development. The method would yield a total (nonviable plus viable) spore count of a surface within minutes and a viable-spore count in about one hour. In this method, spores would be collected from a surface by use of a transparent polymeric tape coated on one side with a polymeric adhesive that would be permeated with one or more reagent(s) for detection of spores by use of visible luminescence. The sticky side of the tape would be pressed against a surface to be assayed, then the tape with captured spores would be placed in a reader that illuminates the sample with ultraviolet light and counts the green luminescence spots under a microscope to quantify the number of bacterial spores per unit area. The visible luminescence spots seen through the microscope would be counted to determine the concentration of spores on the surface. This method is based on the chemical and physical principles of methods described in several prior NASA Tech Briefs articles, including Live/Dead Spore Assay Using DPA-Triggered Tb Luminescence (NPO-30444), Vol. 27, No. 3 (March 2003), page 7a. To recapitulate: The basic idea is to exploit the observations that (1) dipicolinic acid (DPA) is present naturally only in bacterial spores; and (2) when bound to Tb3+ ions, DPA triggers intense green luminescence of the ions under ultraviolet excitation; (3) DPA can be released from the viable spores by using L-alanine to make them germinate; and (4) by autoclaving, microwaving, or sonicating the sample, one can cause all the spores (non-viable as well as viable) to release their DPA. One candidate material for use as the adhesive in the present method is polydimethysiloxane (PDMS). In one variant of the method for obtaining counts of all (viable and nonviable) spores the PDMS would be doped with TbCl3. After collection of a sample, the spores immobilized on the sticky tape surface

Escherichia coli viability determination using dynamic light scattering: a comparison with standard methods.

PubMed

Loske, Achim M; Tello, Elba M; Vargas, Susana; Rodriguez, Rogelio

2014-08-01

To determine the concentration of bacteria in a sample is important in the food industry, medicine and biotechnology. A disadvantage of the plate-counting method is that a microorganism colony could arise from one cell or from many cells. The other standard methodology, known as optical density determination, is based on the turbidity of a suspension and registers all bacteria, dead and alive. In this article, dynamic light scattering is proposed as a fast and reliable method to determine bacterial viability and, consequently, time evolution. Escherichia coli was selected because this microorganism is well known and easy to handle. A correlation between the data from these three techniques was obtained. We were able to calculate the growth rate, usually determined by plate counting or optical density measurement, using dynamic light scattering and to predict bacterial behavior. An analytical relationship between the colony forming units and the light scattered intensity was also deduced.

Spiral Salmonella assay: validation against the standard pour-plate assay.

PubMed

Diehl, M; Fort, F

1996-01-01

The spiral Ames assay, an automated approach to bacterial mutagenicity testing which simplifies the test procedure and reduces the amount of drug required to generate mutagenic dose-response information, has been evaluated and validated for routine screening. The spiral plater delivers the Salmonella bacteria, exogenous metabolic activation system and drug to the surface of a rotating agar plate one on top of another in such a way that a uniform density of bacteria is exposed to a logarithmically decreasing volume of drug. Following an incubation of 48 hr at 37 degrees C, the plates are scanned by a laser counter, and the data are subjected to a computerized analysis. Petri plates of 15 cm diameter were used to provide a concentration range of about 250-fold per plate. The Salmonella were concentrated 20-fold to increase sensitivity. Thirty-eight compounds from a variety of chemical classes, including both pharmaceuticals and known mutagens of moderate to strong potency, were tested in both the spiral and the standard pour-plate assays. There was overall test agreement on positive or negative results for 82% of the compounds tested. When only the results from strains TA98 plus TA100 were considered, the agreement was 87%. When positive results were obtained, the fold increase over vehicle control was on average twice as great for the spiral assay compared to the pour-plate assay. It was concluded that the two assay procedures generally provided comparable results, with the spiral assay being somewhat more sensitive in terms of dose-response than the pour-plate assay.

Changes in bacterial diversity and community structure following pesticides addition to soil estimated by cultivation technique.

PubMed

Cycoń, Mariusz; Piotrowska-Seget, Zofia

2009-07-01

An experiment was conducted under laboratory conditions to investigate the effect of increasing concentrations of fenitrothion (2, 10 and 200 mg a.i./kg soil), diuron (1.5, 7.5 and 150 mg a.i./kg soil) and thiram (3.5, 17.5 and 350 mg a.i./kg soil) on soil respiration, bacterial counts and changes in culturable fraction of soil bacteria. To ascertain these changes, the community structure, bacterial biodiversity and process of colony formation, based on the r/K strategy concept, EP- and CD-indices and the FOR model, respectively, were determined. The results showed that the measured parameters were generally unaffected by the lowest dosages of pesticides, corresponding to the recommended field rates. The highest dosages of fenitrothion and thiram suppressed the peak SIR by 15-70% and 20-80%, respectively, while diuron increased respiration rate by 17-25% during the 28-day experiment. Also, the total numbers of bacteria increased in pesticide-treated soils. However, the reverse effect on day 1 and, in addition, in case of the highest dosages of insecticide on days 14 and 28, was observed. Analysis of the community structure revealed that in all soil treatments bacterial communities were generally dominated by K-strategists. Moreover, differences in the distribution of individual bacteria classes and the gradual domination of bacteria populations belonging to r-strategists during the experiment, as compared to control, was observed. However, on day 1, at the highest pesticide dosages, fast growing bacteria constituted only 1-10% of the total colonies number during 48 h of plate incubation, whereas in remaining samples they reached from 20 to 40% of total cfu. This effect, in case of fenitrothion, lasted till the end of the experiment. At the highest dosages of fenitrothion, diuron and at all dosages of thiram the decrease of biodiversity, as indicated by EP- and CD-indices on day 1, was found. At the next sampling time, no significant retarding or stimulating effect

Bacterial body plans

PubMed Central

Rieger, Tomáš; Neubauer, Zdeněk; Blahůšková, Anna; Cvrčková, Fatima

2008-01-01

The bacterium Serratia marcescens produces a plethora of multicellular shapes of different colorations on solid substrates, allowing immediate visual detection of varieties. Such a plasticity allows studies on multicellular community scale spanning two extremes, from well-elaborated individual colonies to undifferentiated cell mass. For a single strain and medium, we obtained a range of different multicellular bodies, depending on the layout of initial plating. Four principal factors affecting the morphogenetic pathways of such bodies can be distinguished: (1) amount, density and distribution pattern of founder cells; (2) the configuration of surrounding free medium; (3) the presence and character of other bacterial bodies sharing the same niche; and (4) self-perception, resulting in delimitation towards other bodies. The last feature results in an ability of well-formed multicellular individuals to maintain their identity upon a close mutual contact, as well as in spontaneous separation of cell masses in experimental chimeras. We propose an “embryo-like” colony model where multicellular bacterial bodies develop along genuine ontogenetic pathways inherent to the given species (clone), while external shaping forces (like nutrient gradients, pH, etc.,) exert not formative, but only regulative roles in the process. PMID:19513204

Indoor air bacterial load and antibiotic susceptibility pattern of isolates in operating rooms and surgical wards at jimma university specialized hospital, southwest ethiopia.

PubMed

Genet, Chalachew; Kibru, Gebre; Tsegaye, Wondewosen

2011-03-01

Surgical site infection is the second most common health care associated infection. One of the risk factors for such infection is bacterial contamination of operating rooms' and surgical wards' indoor air. In view of that, the microbiological quality of air can be considered as a mirror of the hygienic condition of these rooms. Thus, the objective of this study was to determine the bacterial load and antibiotic susceptibility pattern of isolates in operating rooms' and surgical wards' indoor air of Jimma University Specialized Hospital. A cross sectional study was conducted to measure indoor air microbial quality of operating rooms and surgical wards from October to January 2009/2010 on 108 indoor air samples collected in twelve rounds using purposive sampling technique by Settle Plate Method (Passive Air Sampling following 1/1/1 Schedule). Sample processing and antimicrobial susceptibility testing were done following standard bacteriological techniques. The data was analyzed using SPSS version 16 and interpreted according to scientifically determined baseline values initially suggested by Fisher. The mean aerobic colony counts obtained in OR-1(46cfu/hr) and OR-2(28cfu/hr) was far beyond the set 5-8cfu/hr acceptable standards for passive room. Similarly the highest mean aerobic colony counts of 465cfu/hr and 461cfu/hr were observed in Female room-1 and room-2 respectively when compared to the acceptable range of 250-450cfu/hr. In this study only 3 isolates of S. pyogenes and 48 isolates of S. aureus were identified. Over 66% of S. aureus was identified in Critical Zone of Operating rooms. All isolates of S. aureus showed 100% and 82.8% resistance to methicillin and ampicillin respectively. Higher degree of aerobic bacterial load was measured from operating rooms' and surgical wards' indoor air. Reducing foot trafficking, improving the ventilation system and routine cleaning has to be made to maintain the aerobic bacteria load with in optimal level.

Salivary bacterial leakage into implant-abutment connections: preliminary results of an in vitro study.

PubMed

Mencio, F; Papi, P; Di Carlo, S; Pompa, G

2016-06-01

The occurrence of bacterial leakage in the internal surface of implants, through implant-abutment interface (IAI), is one of the parameters for analyzing the fabrication quality of the connections. The aim of this in vitro study is to evaluate two different types of implant-abutment connections: the screwed connection (Group 1) and the cemented connection (Group 2), analyzing the permeability of the IAI to bacterial colonization, using human saliva as culture medium. A total of twelve implants were tested, six in each experimental group. Five healthy patients were enrolled in this study. Two milliliters of non-stimulated saliva were collected from each subject and mixed in a test tube. After 14 days of incubation of the bacteria sample in the implant fixtures, a PCR-Real Time analysis was performed. Fisher's exact test was used to compare the proportions of implant-abutment assembled structures detected with bacterial leakage. Differences in the bacterial counts of the two groups were compared using the Mann-Whitney U test. A p value < 0.05 was considered significant. The results showed a decreased stability with the screwed implant-abutment connections compared to the cemented implant-abutment connections. A mean total bacterial count of 1.2E+07 (± 0.25E+07) for Group 1 and of 7.2E+04 (± 14.4E+04) for Group 2 was found, with a high level of significance, p = .0001. Within the limitations of this study it can be concluded that bacterial species from human saliva may penetrate along the implant-abutment interface in both connections, however the cemented connection implants showed the lowest amount of bacterial colonization.

Corticosteroids for Bacterial Keratitis

PubMed Central

Srinivasan, Muthiah; Mascarenhas, Jeena; Rajaraman, Revathi; Ravindran, Meenakshi; Lalitha, Prajna; Glidden, David V.; Ray, Kathryn J.; Hong, Kevin C.; Oldenburg, Catherine E.; Lee, Salena M.; Zegans, Michael E.; McLeod, Stephen D.; Lietman, Thomas M.; Acharya, Nisha R.

2013-01-01

Objective To determine whether there is a benefit in clinical outcomes with the use of topical corticosteroids as adjunctive therapy in the treatment of bacterial corneal ulcers. Methods Randomized, placebo-controlled, double-masked, multicenter clinical trial comparing prednisolone sodium phosphate, 1.0%, to placebo as adjunctive therapy for the treatment of bacterial corneal ulcers. Eligible patients had a culture-positive bacterial corneal ulcer and received topical moxifloxacin for at least 48 hours before randomization. Main Outcome Measures The primary outcome was best spectacle-corrected visual acuity (BSCVA) at 3 months from enrollment. Secondary outcomes included infiltrate/scar size, reepithelialization, and corneal perforation. Results Between September 1, 2006, and February 22, 2010, 1769 patients were screened for the trial and 500 patients were enrolled. No significant difference was observed in the 3-month BSCVA (−0.009 logarithm of the minimum angle of resolution [logMAR]; 95% CI, −0.085 to 0.068; P = .82), infiltrate/scar size (P = .40), time to reepithelialization (P = .44), or corneal perforation (P > .99). A significant effect of corticosteroids was observed in subgroups of baseline BSCVA (P = .03) and ulcer location (P = .04). At 3 months, patients with vision of counting fingers or worse at baseline had 0.17 logMAR better visual acuity with corticosteroids (95% CI, −0.31 to −0.02; P = .03) compared with placebo, and patients with ulcers that were completely central at baseline had 0.20 logMAR better visual acuity with corticosteroids (−0.37 to −0.04; P = .02). Conclusions We found no overall difference in 3-month BSCVA and no safety concerns with adjunctive corticosteroid therapy for bacterial corneal ulcers. Application to Clinical Practice Adjunctive topical corticosteroid use does not improve 3-month vision in patients with bacterial corneal ulcers. PMID:21987582

Characterization of the pigment xanthomonadin in the bacterial genus Xanthomonas using micro- and resonance Raman spectroscopy

NASA Astrophysics Data System (ADS)

Paret, Mathews L.; Sharma, Shiv K.; Misra, Anupam K.; Acosta, Tayro; deSilva, Asoka S.; Vowell, Tomie; Alvarez, Anne M.

2012-06-01

We used micro- and resonance Raman spectroscopy with 785 nm and 514.5 nm laser excitation, respectively, to characterize a plant pathogenic bacteria, Xanthomonas axonopodis pv. dieffenbachiae D150. The bacterial genus Xathomonas is closely related to bacterial genus Stenotrophomonas that causes an infection in humans. This study has identified for the first time the unique Raman spectra of the carotenoid-like pigment xanthomonadin of the Xanthomonas strain. Xanthomonadin is a brominated aryl-polyene pigment molecule similar to carotenoids. Further studies were conducted using resonance Raman spectroscopy with 514.5 nm laser excitation on several strains of the bacterial genus Xanthomonas isolated from numerous plants from various geographical locations. The current study revealed that the Raman bands representing the vibrations (v1, v2, v3) of the polyene chain of xanthomonadin are 1003-1005 (v3), 1135-1138 (v2), and 1530 (v1). Overtone bands representing xanthomonadin were identified as 2264-2275 (2v2), and combinational bands at 2653-2662 (v1+ v2). The findings from this study validate our previous finding that the Raman fingerprints of xanthomonadin are unique for the genus Xanthomonas. This facilitates rapid identification (~5 minutes) of Xanthomonas spp. from bacterial culture plates. The xanthomonadin marker is different from Raman markers of many other bacterial genus including Agrobacterium, Bacillus, Clavibacter, Enterobacter, Erwinia, Microbacterium, Paenibacillus, and Ralstonia. This study also identified Xanthomonas spp. from bacterial strains isolated from a diseased wheat sample on a culture plate.

Predictive value of decoy receptor 3 in postoperative nosocomial bacterial meningitis.

PubMed

Liu, Yong-Juan; Shao, Li-Hua; Wang, Qian; Zhang, Jian; Ma, Rui-Ping; Liu, Hai-Hong; Dong, Xiao-Meng; Ma, Li-Xian

2014-11-03

Nosocomial bacterial meningitis requires timely treatment, but what is difficult is the prompt and accurate diagnosis of this disease. The aim of this study was to assess the potential role of decoy receptor 3 (DcR3) levels in the differentiation of bacterial meningitis from non-bacterial meningitis. A total of 123 patients were recruited in this study, among them 80 patients being with bacterial meningitis and 43 patients with non-bacterial meningitis. Bacterial meningitis was confirmed by bacterial culture of cerebrospinal fluid (CSF) culture and enzyme-linked immunosorbent assay (ELISA) was used to detect the level of DcR3 in CSF. CSF levels of DcR3 were statistically significant between patients with bacterial meningitis and those with non-bacterial meningitis (p<0.001). A total of 48.75% of patients with bacterial meningitis received antibiotic>24 h before CSF sampling, which was much higher than that of non-bacterial meningitis. CSF leucocyte count yielded the highest diagnostic value, with an area under the receiver operating characteristic curve (ROC) of 0.928, followed by DcR3. At a critical value of 0.201 ng/mL for DcR3, the sensitivity and specificity were 78.75% and 81.40% respectively. DcR3 in CSF may be a valuable predictor for differentiating patients with bacterial meningitis from those with non-bacterial meningitis. Further studies are needed for the validation of this study.

Predictive Value of Decoy Receptor 3 in Postoperative Nosocomial Bacterial Meningitis

PubMed Central

Liu, Yong-Juan; Shao, Li-Hua; Wang, Qian; Zhang, Jian; Ma, Rui-Ping; Liu, Hai-Hong; Dong, Xiao-Meng; Ma, Li-Xian

2014-01-01

Nosocomial bacterial meningitis requires timely treatment, but what is difficult is the prompt and accurate diagnosis of this disease. The aim of this study was to assess the potential role of decoy receptor 3 (DcR3) levels in the differentiation of bacterial meningitis from non-bacterial meningitis. A total of 123 patients were recruited in this study, among them 80 patients being with bacterial meningitis and 43 patients with non-bacterial meningitis. Bacterial meningitis was confirmed by bacterial culture of cerebrospinal fluid (CSF) culture and enzyme-linked immunosorbent assay (ELISA) was used to detect the level of DcR3 in CSF. CSF levels of DcR3 were statistically significant between patients with bacterial meningitis and those with non-bacterial meningitis (p < 0.001). A total of 48.75% of patients with bacterial meningitis received antibiotic >24 h before CSF sampling, which was much higher than that of non-bacterial meningitis. CSF leucocyte count yielded the highest diagnostic value, with an area under the receiver operating characteristic curve (ROC) of 0.928, followed by DcR3. At a critical value of 0.201 ng/mL for DcR3, the sensitivity and specificity were 78.75% and 81.40% respectively. DcR3 in CSF may be a valuable predictor for differentiating patients with bacterial meningitis from those with non-bacterial meningitis. Further studies are needed for the validation of this study. PMID:25372942

Diversity and dynamics of antibiotic-resistant bacteria in cheese as determined by PCR denaturing gradient gel electrophoresis.

PubMed

Flórez, Ana Belén; Mayo, Baltasar

2015-12-02

This work reports the composition and succession of tetracycline- and erythromycin-resistant bacterial communities in a model cheese, monitored by polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE). Bacterial 16S rRNA genes were examined using this technique to detect structural changes in the cheese microbiota over manufacturing and ripening. Total bacterial genomic DNA, used as a template, was extracted from cultivable bacteria grown without and with tetracycline or erythromycin (both at 25 μg ml(-1)) on a non-selective medium used for enumeration of total and viable cells (Plate Count agar with Milk; PCA-M), and from those grown on selective and/or differential agar media used for counting various bacterial groups; i.e., lactic acid bacteria (de Man, Rogosa and Sharpe agar; MRSA), micrococci and staphylococci (Baird-Parker agar; BPA), and enterobacteria (Violet Red Bile Glucose agar; VRBGA). Large numbers of tetracycline- and erythromycin-resistant bacteria were detected in cheese samples at all stages of ripening. Counts of antibiotic-resistant bacteria varied widely depending on the microbial group and the point of sampling. In general, resistant bacteria were 0.5-1.0 Log10 units fewer in number than the corresponding susceptible bacteria. The PCR-DGGE profiles obtained with DNA isolated from the plates for total bacteria and the different bacterial groups suggested Escherichia coli, Lactococcus lactis, Enterococcus faecalis and Staphylococcus spp. as the microbial types resistant to both antibiotics tested. This study shows the suitability of the PCR-DGGE technique for rapidly identifying and tracking antibiotic resistant populations in cheese and, by extension, in other foods. Copyright © 2015 Elsevier B.V. All rights reserved.

A rapid detection method using flow cytometry to monitor the risk of Legionella in bath water.

PubMed

Taguri, Toshitsugu; Oda, Yasunori; Sugiyama, Kanji; Nishikawa, Toru; Endo, Takuro; Izumiyama, Shinji; Yamazaki, Masayuki; Kura, Fumiaki

2011-07-01

Legionella species are the causative agents of human legionellosis, and bathing facilities have been identified as the sources of infection in several outbreaks in Japan. Researchers in Japan have recently reported evidence of significant associations between bacterial counts and the occurrence of Legionella in bathing facilities and in a hot tub model. A convenient and quantitative bacterial enumeration method is therefore required as an indicator of Legionella contamination or disinfection to replace existing methods such as time-consuming Legionella culture and expensive Legionella-DNA amplification. In this study, we developed a rapid detection method (RDM) to monitor the risk of Legionella using an automated microbial analyzing device based on flow cytometry techniques to measure the total number of bacteria in water samples within two minutes, by detecting typical patterns of scattered light and fluorescence. We first compared the results of our RDM with plate counting results for five filtered hot spring water samples spiked with three species of bacteria, including Legionella. Inactivation of these samples by chlorine was also assessed by the RDM, a live/dead bacterial fluorescence assay and plate counting. Using the RDM, the lower limit of quantitative bacterial counts in the spiked samples was determined as 3.0×10(3)(3.48log)counts mL(-1). We then used a laboratory model of a hot tub and found that the RDM could monitor the growth curve of naturally occurring heterotrophic bacteria with 1 and 2 days' delayed growth of amoeba and Legionella, respectively, and could also determine the killing curve of these bacteria by chlorination. Finally, samples with ≥3.48 or <3.48log total bacterial counts mL(-1) were tested using the RDM from 149 different hot tubs, and were found to be significantly associated with the positive or negative detection of Legionella with 95% sensitivity and 84% specificity. These findings indicated that the RDM can be used for

Bacterial interactions in the rhizosphere of seagrass communities in shallow coastal lagoons.

PubMed

Donnelly, A P; Herbert, R A

1998-12-01

Rooted phanerogam communities in the shallow intertidal and subtidal coastal zone represent productive and healthy ecosystems. Inorganic nutrients are assimilated into seagrass biomass. Much of the organic matter resulting from moribund seagrass is rapidly mineralized, principally by bacteria. The microbial community of the rhizosphere is also highly active due to the supply of organic matter released during photosynthesis. This active sediment community plays an important role through carbon, nitrogen and phosphorous cycling in maintaining the stability and productivity of seagrass meadows. Over the last two decades, however, seagrass meadows in European coastal areas have declined due to increasing pollution. As eutrophication advances a trasition occurs from rooted phanerogram dominated communities to planktonic algal blooms and/or cyanobacterial blooms. Such changes represent the decline of a stable, high biodiversity habitat to an unstable one dominated by a few species. These changes of community structure can occur rapidly once the internal nutrient and organic matter control cycles are exceeded. A field investigation was undertaken to establish the spatial distribution of bacterial populations of Zostera noltii colonized and uncolonized sediment in the Bassin d'Arcachon, France. Bacteria were enumerated using both plate count and MPN techniques for different functional groups as well as determining the total bacterial populations present. Nitrogen fixation, ammonification, sulphate reduction rates, as well as alkaline phosphatase activity were also determined. Colonization of the Z. noltii roots and rhizomes was studied by light and scanning electron microscopy. Results confirmed that higher bacterial populations were present in the rhizosphere of Z. noltii compared to uncolonized sediments. Furthermore, electron microscopy identified the rhizome as the main site of colonization for a diverse range of morphological groups of bacteria. Sulphate reducing

EFFECT OF AEROSOLIZATION ON CULTURABILITY AND VIABILITY OF GRAM-NEGATIVE BACTERIA

EPA Science Inventory

Estimations of the bacterial content of air can be more easily made now than a decade ago, with colony formation the method of choice for enumeration of airborne bacteria.However, plate counts are subject to error because bacteria exposed to the air may remain viable yet lose the...

Adherence of coagulase-negative staphylococci to plastic tissue culture plates: a quantitative model for the adherence of staphylococci to medical devices.

PubMed Central

Christensen, G D; Simpson, W A; Younger, J J; Baddour, L M; Barrett, F F; Melton, D M; Beachey, E H

1985-01-01

The adherence of coagulase-negative staphylococci to smooth surfaces was assayed by measuring the optical densities of stained bacterial films adherent to the floors of plastic tissue culture plates. The optical densities correlated with the weight of the adherent bacterial film (r = 0.906; P less than 0.01). The measurements also agreed with visual assessments of bacterial adherence to culture tubes, microtiter plates, and tissue culture plates. Selected clinical strains were passed through a mouse model for foreign body infections and a rat model for catheter-induced endocarditis. The adherence measurements of animal passed strains remained the same as those of the laboratory-maintained parent strain. Spectrophotometric classification of coagulase-negative staphylococci into nonadherent and adherent categories according to these measurements had a sensitivity, specificity, and accuracy of 90.6, 80.8, and 88.4%, respectively. We examined a previously described collection of 127 strains of coagulase-negative staphylococci isolated from an outbreak of intravascular catheter-associated sepsis; strains associated with sepsis were more adherent than blood culture contaminants and cutaneous strains (P less than 0.001). We also examined a collection of 84 strains isolated from pediatric patients with cerebrospinal fluid (CSF) shunts; once again, pathogenic strains were more adherent than were CSF contaminants (P less than 0.01). Finally, we measured the adherence of seven endocarditis strains. As opposed to strains associated with intravascular catheters and CSF shunts, endocarditis strains were less adherent than were saprophytic strains of coagulase-negative staphylococci. The optical densities of bacterial films adherent to plastic tissue culture plates serve as a quantitative model for the study of the adherence of coagulase-negative staphylococci to medical devices, a process which may be important in the pathogenesis of foreign body infections. Images PMID:3905855

Biodegradable Chitosan Coating Incorporated with Black Pepper Essential Oil for Shelf Life Extension of Common Carp (Cyprinus carpio) during Refrigerated Storage.

PubMed

Moosavi-Nasab, Marzieh; Shad, Ehsan; Ziaee, Esmaeil; Yousefabad, Seyyed Hossein Asadi; Golmakani, Mohammad Taghi; Azizinia, Mehdi

2016-06-01

Chitosan (Ch) coating incorporated with black pepper essential oil (Ch+BPEO) was studied to extend the shelf life of common carp (Cyprinus carpio) during refrigerated storage at 4 ± 1°C. The chemical composition of BPEO was characterized using gas chromatography-mass spectrometry (GC-MS). Antibacterial properties of BPEO were determined by disk diffusion agar, MIC, and MBC. Ch (2% [wt/vol]) and Ch+BPEO (2% [wt/vol] Ch with 1.5% [vol/vol] BPEO) were used for common carp fillet coating. The samples were analyzed periodically for chemical (pH, total volatile basic nitrogen) and microbiological (aerobic plate count, psychrophilic bacteria count, lactic acid bacteria, and Enterobacteriaceae bacterial counts) characteristics during 16 days. The GC-MS results indicated that main components in BPEO were carene, caryophyllene, limonene, β-pinene, and α-pinene. The samples coated with Ch and Ch+BPEO resulted in lower pH and total volatile basic nitrogen values in comparison with the control. The microbiological analysis of fish fillets during refrigerated storage clearly indicated that Ch+BPEO coating significantly reduced the fish fillet microbial load. The aerobic plate count, psychrophilic bacteria count, lactic acid bacteria count, and Enterobacteriaceae bacterial count of samples coated with Ch+BPEO were reduced approximately 4.1, 3.9, 2.3, and 2.8 log CFU/g, respectively, at the end of the storage period. Finally, Ch and Ch+BPEO effectively improved the quality of fish fillet during refrigerated storage and extended the shelf life of fish fillets from 8 to 16 days. Black pepper; Chitosan; Common carp; Essential oil.

Spore-forming organisms in platelet concentrates: a challenge in transfusion bacterial safety.

PubMed

Störmer, M; Vollmer, T; Kleesiek, K; Dreier, J

2008-12-01

Bacterial detection and pathogen reduction are widely used methods of minimizing the risk of transfusion-transmitted bacterial infection. But, bacterial spores are highly resistant to chemical and physical agents. In this study, we assessed the bacterial proliferation of spore-forming organisms seeded into platelet concentrates (PCs) to demonstrate that spores can enter the vegetative state in PCs during storage. In the in vitro study, PCs were inoculated with 1-10 spores mL(-1)of Bacillus cereus (n = 1), Bacillus subtilis (n = 2) and Clostridium sporogenes (n = 2). Sampling was performed during 6-day aerobic storage at 22 degrees C. The presence of bacteria was assessed by plating culture, automated culture and real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Spores of the C. sporogenes do not enter the vegetative phase under PC storage conditions, whereas B. subtilis and B. cereus showed growth in the PC and could be detected using RT-PCR and automated culture. Depending on the species and inoculums, bacterial spores may enter the vegetative phase during PC storage and can be detected by bacterial detection methods.

Host Biomarkers for Distinguishing Bacterial from Non-Bacterial Causes of Acute Febrile Illness: A Comprehensive Review

PubMed Central

Kapasi, Anokhi J.; Dittrich, Sabine; González, Iveth J.; Rodwell, Timothy C.

2016-01-01

Background In resource limited settings acute febrile illnesses are often treated empirically due to a lack of reliable, rapid point-of-care diagnostics. This contributes to the indiscriminate use of antimicrobial drugs and poor treatment outcomes. The aim of this comprehensive review was to summarize the diagnostic performance of host biomarkers capable of differentiating bacterial from non-bacterial infections to guide the use of antibiotics. Methods Online databases of published literature were searched from January 2010 through April 2015. English language studies that evaluated the performance of one or more host biomarker in differentiating bacterial from non-bacterial infection in patients were included. Key information extracted included author information, study methods, population, pathogens, clinical information, and biomarker performance data. Study quality was assessed using a combination of validated criteria from the QUADAS and Lijmer checklists. Biomarkers were categorized as hematologic factors, inflammatory molecules, cytokines, cell surface or metabolic markers, other host biomarkers, host transcripts, clinical biometrics, and combinations of markers. Findings Of the 193 citations identified, 59 studies that evaluated over 112 host biomarkers were selected. Most studies involved patient populations from high-income countries, while 19% involved populations from low- and middle-income countries. The most frequently evaluated host biomarkers were C-reactive protein (61%), white blood cell count (44%) and procalcitonin (34%). Study quality scores ranged from 23.1% to 92.3%. There were 9 high performance host biomarkers or combinations, with sensitivity and specificity of ≥85% or either sensitivity or specificity was reported to be 100%. Five host biomarkers were considered weak markers as they lacked statistically significant performance in discriminating between bacterial and non-bacterial infections. Discussion This manuscript provides a summary

Direct viable count as test for toxicity assessment: the effects of four metals on a Salmonella enteritidis strain.

PubMed

Scoglio, M E; Di Pietro, A; Anzalone, C; Calimeri, S; Lo Giudice, D; Trimarchi, G R

2000-01-01

The toxicity of synthetic sewage containing increasing concentrations of arsenic (.125, .25, .5, 1.0 mg L-1), cadmium (.02, .05, .1, .2 mg L-1), lead (.2, .5, 1.0, 2.0 mg L-1) and nickel (.5, 1.0, 2.0, 4.0 mg L-1) has been investigated by determining the total direct count (TDC) and the direct viable count (DVC) of Salmonella enteritidis by means of an immunofluorescence technique (IFA). This has been done in order to evaluate the possibility of using the IFA technique to estimate the toxicity of complex effluents. Arsenic, cadmium and nickel produced a concentration-dependent reduction in the number of viable bacterial cells. This was more clear when the viable bacterial cells were considered than when only the culturable part was used. Lead did not show a concentration-dependent and reproducible effect. At the highest concentrations allowed by the Italian wastewater regulations, lead, cadmium, arsenic and nickel reduced the viable/total bacterial cells ratio to 74.5%, 68.5%, 28.4% and 6.9%, respectively. The toxic effects of the metals were also tested using the standard Microtox assay.

Hydrodynamics of a flexible plate between pitching rigid plates

NASA Astrophysics Data System (ADS)

Kim, Junyoung; Kim, Daegyoum

2017-11-01

The dynamics of a flexible plate have been studied as a model problem in swimming and flying of animals and fluid-structure interaction of plants and flags. Motivated by fish schooling and an array of sea grasses, we investigate the dynamics of a flexible plate closely placed between two pitching rigid plates. In most studies on passive deformation of the flexible plate, the plate is immersed in a uniform flow or a wavy flow. However, in this study, the flexible plate experiences periodic deformation by the oscillatory flow generated by the prescribed pitching motion of the rigid plates. In our model, the pitching axes of the rigid plates and the clamping position of the flexible plate are aligned on the same line. The flexible plate shows various responses depending on length and pitching frequency of rigid plates, thickness of a flexible plate, and free-stream velocity. To find the effect of each variable on the response of the flexible plate, amplitude of a trailing edge and modal contribution of a flapping motion are compared, and flow structure around the flexible plate is examined.

Characterisation of Arctic Bacterial Communities in the Air above Svalbard

PubMed Central

Cuthbertson, Lewis; Amores-Arrocha, Herminia; Malard, Lucie A.; Els, Nora; Sattler, Birgit; Pearce, David A.

2017-01-01

Atmospheric dispersal of bacteria is increasingly acknowledged as an important factor influencing bacterial community biodiversity, biogeography and bacteria–human interactions, including those linked to human health. However, knowledge about patterns in microbial aerobiology is still relatively scarce, and this can be attributed, in part, to a lack of consensus on appropriate sampling and analytical methodology. In this study, three different methods were used to investigate aerial biodiversity over Svalbard: impaction, membrane filtration and drop plates. Sites around Svalbard were selected due to their relatively remote location, low human population, geographical location with respect to air movement and the tradition and history of scientific investigation on the archipelago, ensuring the presence of existing research infrastructure. The aerial bacterial biodiversity found was similar to that described in other aerobiological studies from both polar and non-polar environments, with Proteobacteria, Actinobacteria, and Firmicutes being the predominant groups. Twelve different phyla were detected in the air collected above Svalbard, although the diversity was considerably lower than in urban environments elsewhere. However, only 58 of 196 bacterial genera detected were consistently present, suggesting potentially higher levels of heterogeneity. Viable bacteria were present at all sampling locations, showing that living bacteria are ubiquitous in the air around Svalbard. Sampling location influenced the results obtained, as did sampling method. Specifically, impaction with a Sartorius MD8 produced a significantly higher number of viable colony forming units (CFUs) than drop plates alone. PMID:28481257

Microorganisms as an Indicator of Hygiene Status Among Migrant Food Handlers in Peninsular Malaysia.

PubMed

Woh, Pei Yee; Thong, Kwai Lin; Lim, Yvonne Ai Lian; Behnke, Jerzy Marian; Lewis, John Watkin; Mohd Zain, Siti Nursheena

2017-10-01

This study used microbial indicators to assess the hygiene status of 383 migrant food handlers from 3 urban cities in Peninsular Malaysia. Microbiological analysis revealed that all the hand swabs tested 99.5% positive for aerobic plate counts (mean [M] ± standard deviation [SD] = 3.57 ± 0.83 log 10 CFU [colony forming unit]), 20.8% positive for total coliform/ Escherichia coli (M ± SD = 0.30 ± 0.67 log 10 CFU), and 63.4% positive for Staphylococcus aureus (M ± SD = 1.38 ± 1.26 log 10 CFU). In addition, aerobic plate counts and Staphylococcus aureus counts exceeded the acceptable standard levels. Bacterial counts were found to be significantly associated with subjects' country of origin ( P = .019) and working responsibilities ( P = .001). Our findings indicate high probability of transmission of pathogenic bacteria from the food handlers' hands to customers during meal preparation and serving. This calls for improvements in personal hygiene and sanitation standards by the relevant health authorities among migrant food handlers.

Influence of a Brazilian wild green propolis on the enamel mineral loss and Streptococcus mutans' count in dental biofilm.

PubMed

Cardoso, Julia Gabiroboertz; Iorio, Natalia Lopes Pontes; Rodrigues, Luís Fernando; Couri, Maria Luiza Barra; Farah, Adriana; Maia, Lucianne Cople; Antonio, Andréa Gonçalves

2016-05-01

This study investigated the anti-demineralizing and antibacterial effects of a propolis ethanolic extract (EEP) against Streptococcus mutans dental biofilm. Blocks of sound bovine enamel (n=24) were fixed on polystyrene plates. S. mutans inoculum (ATCC 25175) and culture media were added (48 h-37 °C) to form biofilm. Blocks with biofilm received daily treatment (30 μL/1 min), for 5 days, as following: G1 (EEP 33.3%); G2 (chlorhexidine digluconate 0.12%); G3 (ethanol 80%); and G4 (Milli-Q water). G5 and G6 were blocks without biofilm that received only EEP and Milli-Q water, respectively. Final surface hardness was evaluated and the percentage of hardness loss (%HL) was calculated. The EEP extract pH and total solids were determined. S. mutans count was expressed by log10 scale of Colony-Forming Units (CFU/mL). One way ANOVA was used to compare results which differed at a 95% significance level. G2 presented the lowest average %HL value (68.44% ± 12.98) (p=0.010), while G4 presented the highest (90.49% ± 5.38%HL) (p=0.007). G1 showed %HL (84.41% ± 2.77) similar to G3 (87.80% ± 6.89) (p=0.477). Groups G5 and G6 presented %HL=16.11% ± 7.92 and 20.55% ± 10.65; respectively (p=0.952). G1 and G4 differed as regards to S. mutans count: 7.26 ± 0.08 and 8.29 ± 0.17 CFU/mL, respectively (p=0.001). The lowest bacterial count was observed in chlorhexidine group (G2=6.79 ± 0.10 CFU/mL) (p=0.043). There was no difference between S. mutans count of G3 and G4 (p=0.435). The EEP showed pH 4.8 and total soluble solids content=25.9 Brix. The EEP seems to be a potent antibacterial substance against S. mutans dental biofilm, but presented no inhibitory action on the de-remineralization of caries process. Copyright © 2016 Elsevier Ltd. All rights reserved.

The effect of a honey based gel and silver sulphadiazine on bacterial infections of in vitro burn wounds.

PubMed

Boekema, B K H L; Pool, L; Ulrich, M M W

2013-06-01

Bacterial contamination remains a constant threat in burn wound care. Topical treatments to combat contaminations have good bactericidal effects but can have detrimental effects for the healing process. Treatments with for example silver can increase healing times. Honey based products can be a good alternative as it is antibacterial and patient-friendly. We evaluated the bactericidal and cytotoxic effects of a honey based gel and silver sulphadiazine in a human burn wound model with Pseudomonas aeruginosa. After adding 10(5)colony forming units of P. aeruginosa, topical treatments were applied on the burn wound models. After 2, 12, 24, 28 and 70 h, bacteria were dislodged and counted by plating dilutions. Cytotoxic effects were evaluated histologically in samples of burn wound models treated topically for 3 weeks, without bacteria. L-Mesitran Soft significantly reduced the bacterial load (5-log reduction) up to 24h but did not completely eliminate bacteria from the burn wounds. After Flammazine(®) treatment, only a few colony forming units were observed at all time points. In contrast, re-epithelialization was significantly reduced after application of Flammazine(®) compared to L-Mesitran Soft or control. This in vitro model of burn wound infection can be used to evaluate topical treatments. L-Mesitran Soft is a good alternative for treating burn wounds but the slightly lower bactericidal activity in the burn wound model warrants a higher frequency of application. Copyright © 2012 Elsevier Ltd and ISBI. All rights reserved.

Analysis of Bacterial Community Structure in Sulfurous-Oil-Containing Soils and Detection of Species Carrying Dibenzothiophene Desulfurization (dsz) Genes

PubMed Central

Duarte, Gabriela Frois; Rosado, Alexandre Soares; Seldin, Lucy; de Araujo, Welington; van Elsas, Jan Dirk

2001-01-01

The selective effects of sulfur-containing hydrocarbons, with respect to changes in bacterial community structure and selection of desulfurizing organisms and genes, were studied in soil. Samples taken from a polluted field soil (A) along a concentration gradient of sulfurous oil and from soil microcosms treated with dibenzothiophene (DBT)-containing petroleum (FSL soil) were analyzed. Analyses included plate counts of total bacteria and of DBT utilizers, molecular community profiling via soil DNA-based PCR-denaturing gradient gel electrophoresis (PCR-DGGE), and detection of genes that encode enzymes involved in the desulfurization of hydrocarbons, i.e., dszA, dszB, and dszC.Data obtained from the A soil showed no discriminating effects of oil levels on the culturable bacterial numbers on either medium used. Generally, counts of DBT degraders were 10- to 100-fold lower than the total culturable counts. However, PCR-DGGE showed that the numbers of bands detected in the molecular community profiles decreased with increasing oil content of the soil. Analysis of the sequences of three prominent bands of the profiles generated with the highly polluted soil samples suggested that the underlying organisms were related to Actinomyces sp., Arthrobacter sp., and a bacterium of uncertain affiliation. dszA, dszB, and dszC genes were present in all A soil samples, whereas a range of unpolluted soils gave negative results in this analysis. Results from the study of FSL soil revealed minor effects of the petroleum-DBT treatment on culturable bacterial numbers and clear effects on the DBT-utilizing communities. The molecular community profiles were largely stable over time in the untreated soil, whereas they showed a progressive change over time following treatment with DBT-containing petroleum. Direct PCR assessment revealed the presence of dszB-related signals in the untreated FSL soil and the apparent selection of dszA- and dszC-related sequences by the petroleum-DBT treatment

Silver-nanoparticle-coated biliary stent inhibits bacterial adhesion in bacterial cholangitis in swine.

PubMed

Wen, Wei; Ma, Li-Mei; He, Wei; Tang, Xiao-Wei; Zhang, Yin; Wang, Xiang; Liu, Li; Fan, Zhi-Ning

2016-02-01

One of the major limitations of biliary stents is the stent occlusion, which is closely related to the over-growth of bacteria. This study aimed to evaluate the feasibility of a novel silver-nanoparticle-coated polyurethane (Ag/PU) stent in bacterial cholangitis model in swine. Ag/PU was designed by coating silver nanoparticles on polyurethane (PU) stent. Twenty-four healthy pigs with bacterial cholangitis using Ag/PU and PU stents were randomly divided into an Ag/PU stent group (n=12) and a PU stent group (n=12), respectively. The stents were inserted by standard endoscopic retrograde cholangiopancreatography. Laboratory assay was performed for white blood cell (WBC) count, alanine aminotransferase (ALT), interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) at baseline time, 8 hours, 1, 2, 3, and 7 days after stent placements. The segment of bile duct containing the stent was examined histologically ex vivo. Implanted biliary stents were examined by a scan electron microscope. The amount of silver release was also measured in vitro. The number of inflammatory cells and level of ALT, IL-1beta and TNF-alpha were significantly lower in the Ag/PU stent group than in the PU stent group. Hyperplasia of the mucosa was more severe in the PU stent group than in the Ag/PU stent group. In contrast to the biofilm of bacteria on the PU stent, fewer bacteria adhered to the Ag/PU stent. PU biliary stents modified with silver nanoparticles are able to alleviate the inflammation of pigs with bacterial cholangitis. Silver-nanoparticle-coated stents are resistant to bacterial adhesion.

Effect of mixing techniques on bacterial attachment and disinfection time of polyether impression material

PubMed Central

Guler, Umut; Budak, Yasemin; Ruh, Emrah; Ocal, Yesim; Canay, Senay; Akyon, Yakut

2013-01-01

Objective: The aim of this study was 2-fold. The first aim was to evaluate the effects of mixing technique (hand-mixing or auto-mixing) on bacterial attachment to polyether impression materials. The second aim was to determine whether bacterial attachment to these materials was affected by length of exposure to disinfection solutions. Materials and Methods: Polyether impression material samples (n = 144) were prepared by hand-mixing or auto-mixing. Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa were used in testing. After incubation, the bacterial colonies were counted and then disinfectant solution was applied. The effect of disinfection solution was evaluated just after the polymerization of impression material and 30 min after polymerization. Differences in adherence of bacteria to the samples prepared by hand-mixing and to those prepared by auto-mixing were assessed by Kruskal-Wallis and Mann-Whitney U-tests. For evaluating the efficiency of the disinfectant, Kruskal-Wallis multiple comparisons test was used. Results: E. coli counts were higher in hand-mixed materials (P < 0.05); no other statistically significant differences were found between hand- and auto-mixed materials. According to the Kruskal-Wallis test, significant differences were found between the disinfection procedures (Z > 2.394). Conclusion: The methods used for mixing polyether impression material did not affect bacterial attachment to impression surfaces. In contrast, the disinfection procedure greatly affects decontamination of the impression surface. PMID:24966729

Effect of mixing techniques on bacterial attachment and disinfection time of polyether impression material.

PubMed

Guler, Umut; Budak, Yasemin; Ruh, Emrah; Ocal, Yesim; Canay, Senay; Akyon, Yakut

2013-09-01

The aim of this study was 2-fold. The first aim was to evaluate the effects of mixing technique (hand-mixing or auto-mixing) on bacterial attachment to polyether impression materials. The second aim was to determine whether bacterial attachment to these materials was affected by length of exposure to disinfection solutions. Polyether impression material samples (n = 144) were prepared by hand-mixing or auto-mixing. Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa were used in testing. After incubation, the bacterial colonies were counted and then disinfectant solution was applied. The effect of disinfection solution was evaluated just after the polymerization of impression material and 30 min after polymerization. Differences in adherence of bacteria to the samples prepared by hand-mixing and to those prepared by auto-mixing were assessed by Kruskal-Wallis and Mann-Whitney U-tests. For evaluating the efficiency of the disinfectant, Kruskal-Wallis multiple comparisons test was used. E. coli counts were higher in hand-mixed materials (P < 0.05); no other statistically significant differences were found between hand- and auto-mixed materials. According to the Kruskal-Wallis test, significant differences were found between the disinfection procedures (Z > 2.394). The methods used for mixing polyether impression material did not affect bacterial attachment to impression surfaces. In contrast, the disinfection procedure greatly affects decontamination of the impression surface.

Compton suppression gamma-counting: The effect of count rate

USGS Publications Warehouse

Millard, H.T.

1984-01-01

Past research has shown that anti-coincidence shielded Ge(Li) spectrometers enhanced the signal-to-background ratios for gamma-photopeaks, which are situated on high Compton backgrounds. Ordinarily, an anti- or non-coincidence spectrum (A) and a coincidence spectrum (C) are collected simultaneously with these systems. To be useful in neutron activation analysis (NAA), the fractions of the photopeak counts routed to the two spectra must be constant from sample to sample to variations must be corrected quantitatively. Most Compton suppression counting has been done at low count rate, but in NAA applications, count rates may be much higher. To operate over the wider dynamic range, the effect of count rate on the ratio of the photopeak counts in the two spectra (A/C) was studied. It was found that as the count rate increases, A/C decreases for gammas not coincident with other gammas from the same decay. For gammas coincident with other gammas, A/C increases to a maximum and then decreases. These results suggest that calibration curves are required to correct photopeak areas so quantitative data can be obtained at higher count rates. ?? 1984.

LENGTH-HETEROGENEITY POLYMERASE CHAIN REACTION (LH-PCR) AS AN INDICATOR OF STREAM SANITARY AND ECOLOGICAL CONDITION: OPTIMAL SAMPLE SIZE AND HOLDING CONDITIONS

EPA Science Inventory

The use of coliform plate count data to assess stream sanitary and ecological condition is limited by the need to store samples at 4oC and analyze them within a 24-hour period. We are testing LH-PCR as an alternative tool to assess the bacterial load of streams, offering a cost ...

Okara: A Nutritionally Valuable By-product Able to Stabilize Lactobacillus plantarum during Freeze-drying, Spray-drying, and Storage.

PubMed

Quintana, Gabriel; Gerbino, Esteban; Gómez-Zavaglia, Andrea

2017-01-01

Okara is a nutritionally valuable by-product produced in large quantities as result of soymilk elaboration. This work proposes its use as both culture and dehydration medium during freeze-drying, spray-drying, and storage of Lactobacillus plantarum CIDCA 83114. Whole and defatted okara were employed as culture media for L. plantarum CIDCA 83114. The growth kinetics were followed by plate counting and compared with those of bacteria grown in MRS broth (control). No significant differences in plate counting were observed in the three media. The fatty acid composition of bacteria grown in whole and defatted okara showed a noticeable increase in the unsaturated/saturated (U/S) fatty acid ratio, with regard to bacteria grown in MRS. This change was mainly due to the increase in polyunsaturated fatty acids, namely C18:2. For dehydration assays, cultures in the stationary phase were neutralized and freeze-dried (with or without the addition of 250 mM sucrose) or spray-dried. Bacteria were plate counted immediately after freeze-drying or spray-drying and during storage at 4°C for 90 days. Freeze-drying in whole okara conducted to the highest bacterial recovery. Regarding storage, spray-dried bacteria previously grown in whole and defatted okara showed higher plate counts than those grown in MRS. On the contrary, freeze-dried bacteria previously grown in all the three culture media were those with the lowest plate counts. The addition of sucrose to the dehydration media improved their recovery. The higher recovery of microorganisms grown in okara after freeze-drying and spray-drying processes and during storage was ascribed to both the presence of fiber and proteins in the dehydration media, and the increase in U/S fatty acids ratio in bacterial membranes. The obtained results support for the first time the use of okara as an innovative matrix to deliver L. plantarum . Considering that okara is an agro-waste obtained in large quantities, these results represent an

Okara: A Nutritionally Valuable By-product Able to Stabilize Lactobacillus plantarum during Freeze-drying, Spray-drying, and Storage

PubMed Central

Quintana, Gabriel; Gerbino, Esteban; Gómez-Zavaglia, Andrea

2017-01-01

Okara is a nutritionally valuable by-product produced in large quantities as result of soymilk elaboration. This work proposes its use as both culture and dehydration medium during freeze-drying, spray-drying, and storage of Lactobacillus plantarum CIDCA 83114. Whole and defatted okara were employed as culture media for L. plantarum CIDCA 83114. The growth kinetics were followed by plate counting and compared with those of bacteria grown in MRS broth (control). No significant differences in plate counting were observed in the three media. The fatty acid composition of bacteria grown in whole and defatted okara showed a noticeable increase in the unsaturated/saturated (U/S) fatty acid ratio, with regard to bacteria grown in MRS. This change was mainly due to the increase in polyunsaturated fatty acids, namely C18:2. For dehydration assays, cultures in the stationary phase were neutralized and freeze-dried (with or without the addition of 250 mM sucrose) or spray-dried. Bacteria were plate counted immediately after freeze-drying or spray-drying and during storage at 4°C for 90 days. Freeze-drying in whole okara conducted to the highest bacterial recovery. Regarding storage, spray-dried bacteria previously grown in whole and defatted okara showed higher plate counts than those grown in MRS. On the contrary, freeze-dried bacteria previously grown in all the three culture media were those with the lowest plate counts. The addition of sucrose to the dehydration media improved their recovery. The higher recovery of microorganisms grown in okara after freeze-drying and spray-drying processes and during storage was ascribed to both the presence of fiber and proteins in the dehydration media, and the increase in U/S fatty acids ratio in bacterial membranes. The obtained results support for the first time the use of okara as an innovative matrix to deliver L. plantarum. Considering that okara is an agro-waste obtained in large quantities, these results represent an

Distinctive colonization of Bacillus sp. bacteria and the influence of the bacterial biofilm on electrochemical behaviors of aluminum coatings.

PubMed

Abdoli, Leila; Suo, Xinkun; Li, Hua

2016-09-01

Formation of biofilm is usually essential for the development of biofouling and crucially impacts the corrosion of marine structures. Here we report the attachment behaviors of Bacillus sp. bacteria and subsequent formation of bacterial biofilm on stainless steel and thermal sprayed aluminum coatings in artificial seawater. The colonized bacteria accelerate the corrosion of the steel plates, and markedly enhance the anti-corrosion performances of the Al coatings in early growth stage of the bacterial biofilm. After 7days incubation, the biofilm formed on the steel is heterogeneous while exhibits homogeneous feature on the Al coating. Atomic force microscopy examination discloses inception of formation of local pitting on steel plates associated with significantly roughened surface. Electrochemical testing suggests that the impact of the bacterial biofilm on the corrosion behaviors of marine structures is not decided by the biofilm alone, it is instead attributed to synergistic influence by both the biofilm and physicochemical characteristics of the substratum materials. Copyright © 2016 Elsevier B.V. All rights reserved.

Assessment of the bacteriological activity associated with granular activated carbon treatment of drinking water.

PubMed Central

Stewart, M H; Wolfe, R L; Means, E G

1990-01-01

Bacteriological analyses were performed on the effluent from a conventional water treatment pilot plant in which granular activated carbon (GAC) had been used as the final process to assess the impact of GAC on the microbial quality of the water produced. Samples were collected twice weekly for 160 days from the effluents of six GAC columns, each of which used one of four different empty-bed contact times (7.5, 15, 30, and 60 min). The samples were analyzed for heterotrophic plate counts and total coliforms. Effluent samples were also exposed to chloramines and free chlorine for 60 min (pH 8.2, 23 degrees C). Bacterial identifications were performed on the disinfected and nondisinfected effluents. Additional studies were conducted to assess the bacteriological activity associated with released GAC particles. The results indicated that heterotrophic plate counts in the effluents from all columns increased to 10(5) CFU/ml within 5 days and subsequently stabilized at 10(4) CFU/ml. The heterotrophic plate counts did not differ at different empty-bed contact times. Coliforms (identified as Enterobacter spp.) were recovered from the nondisinfected effluent on only two occasions. The disinfection results indicated that 1.5 mg of chloramines per liter inactivated approximately 50% more bacteria than did 1.0 mg of free chlorine per liter after 1 h of contact time. Chloramines and chlorine selected for the development of different bacterial species--Pseudomonas spp. and Flavobacterium spp., respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2082828

Contribution of bacterial cells to lacustrine organic matter based on amino sugars and D-amino acids

NASA Astrophysics Data System (ADS)

Carstens, Dörte; Köllner, Krista E.; Bürgmann, Helmut; Wehrli, Bernhard; Schubert, Carsten J.

2012-07-01

Amino sugars (ASs), D-amino acids (D-AAs), and bacterial cell counts were measured in two Swiss lakes to study the contribution of bacterial cells to organic matter (OM) and the fate of ASs and bacterial amino biomarkers during OM degradation. Concentrations of individual ASs (glucosamine, galactosamine, muramic acid, and mannosamine) in the particulate and total OM pools were analyzed in water-column profiles of Lake Brienz (oligotrophic and oxic throughout the entire water column) and Lake Zug (eutrophic, stratified, and permanently anoxic below 170 m) in spring and in fall. Generally, carbon-normalized AS concentrations decreased with water depth, indicating the preferential decomposition of ASs. For Lake Brienz the relative loss of particulate ASs was higher than in Lake Zug, suggesting enhanced AS turnover in an oligotrophic environment. AS ratio changes in the water column revealed a replacement of plankton biomass with OM from heterotrophic microorganisms with increasing water depth. Similar to the ASs, highest carbon normalized D-AA concentrations were found in the upper water column with decreasing concentrations with depth and an increase close to the sediments. In Lake Zug, an increase in the percentage of D-AAs also showed the involvement of bacteria in OM degradation. Estimations of OM derived from bacterial cells using cell counts and the bacterial biomarkers muramic acid and D-AAs gave similar results. For Lake Brienz 0.2-14% of the organic carbon pool originated from bacterial cells, compared to only 0.1-5% in Lake Zug. Based on our estimates, muramic acid appeared primarily associated with bacterial biomass and not with refractory bacterial necromass. Our study underscores that bacteria are not only important drivers of OM degradation in lacustrine systems, they also represent a significant source of OM themselves, especially in oligotrophic lakes.

Influence of Biopreparations on the Bacterial Community of Oily Waste

NASA Astrophysics Data System (ADS)

Biktasheva, L. R.; Galitskaya, P. Yu; Selivanovskaya, S. Yu

2018-01-01

Oil pollution is reported to be one the most serious environmental problems nowadays. Therefore, methods of remediation of oily polluted soils and oily wastes are of great importance. Bioremediation being a perspective method of sanitation of oil pollutions, includes biostimulation of the polluted sites’ indigenous microflora, and in some cases additional introduction of active strains able to decompose hydrocarbon. The efficacy of introducing such biopreparations depends on the interactions between the introduced microbes and the indigenous ones. In this study, the influence of bacterial consortium (Rhodococcus jialingiae, Stenotrophomonas rhizophila and Pseudomonas gessardii) introduction on the bioremediation of an oily waste sampled from a refinery situated in the Mari El region (Russia) was estimated. Single and multiple inoculations of the consortium in addition to moistening and aeration were compared with a control sample, which included only aeration and moistening of the waste. It was shown, that two of the three introduced strains (Rh. jialingiae and Ps.gessardii) gene copy numbers were higher in the inoculated variants than in the control sample and with their initial counts, which meant that these strains survived and included into the bacterial community of the wastes. At the same time, bacterial counts were significantly lower, and the physiological profile of waste microflora slightly altered in the inoculated remediation variants as compared with the control sample. Interestingly, no difference in the degradation rates of hydrocarbons was revealed in the inoculated remediation variants and the control sample.

New Bacterial Infection in the Prostate after Transrectal Prostate Biopsy.

PubMed

Seo, Yumi; Lee, Gilho

2018-04-23

The prostate is prone to infections. Hypothetically, bacteria can be inoculated into the prostate during a transrectal prostate biopsy (TRPB) and progress into chronic bacterial prostatitis. Therefore, we examined new bacterial infections in biopsied prostates after TRPB and whether they affect clinical characteristics in the biopsied patients. Of men whose prostate cultures have been taken prior to TRPB, 105 men with bacteria-free benign prostate pathology underwent an additional repeated prostate culture within a year after TRPB. Twenty out of 105 men (19.05%) acquired new bacteria in their naïve prostates after TRPB. Except for one single case of Escherichia coli infection, 19 men had acquired gram-positive bacteria species. Between the culture-positive and negative groups, there were no significant differences in age, serum prostate-specific antigen (PSA) level, white blood cell (WBC) counts in expressed prostatic secretion (EPS), prostate volume, symptom severities in Korean version of the National Institutes of Health-Chronic Prostatitis Symptom Index (NIH-CPSI) questionnaire, and patient-specific risk factors for biopsy associated infectious complications. Additionally, the TRPB procedure increased the WBC counts in post-biopsy EPS ( P = 0.031, McNemar test), but did not increase the serum PSA level and symptoms of NIH-CPSI in 20 men who acquired new bacteria after TRPB. The TRPB procedure was significantly associated with acquiring new bacterial infections in the biopsied prostate, but these localized bacteria did not affect patients' serum PSA level and symptoms after biopsy.

Covalent Immobilization of Enoxacin onto Titanium Implant Surfaces for Inhibiting Multiple Bacterial Species Infection and In Vivo Methicillin-Resistant Staphylococcus aureus Infection Prophylaxis

PubMed Central

Nie, Bin'en; Long, Teng; Ao, Haiyong; Zhou, Jianliang; Tang, Tingting

2016-01-01

ABSTRACT Infection is one of the most important causes of titanium implant failure in vivo. A developing prophylactic method involves the immobilization of antibiotics, especially vancomycin, onto the surface of the titanium implant. However, these methods have a limited effect in curbing multiple bacterial infections due to antibiotic specificity. In the current study, enoxacin was covalently bound to an amine-functionalized Ti surface by use of a polyethylene glycol (PEG) spacer, and the bactericidal effectiveness was investigated in vitro and in vivo. The titanium surface was amine functionalized with 3-aminopropyltriethoxysilane (APTES), through which PEG spacer molecules were covalently immobilized onto the titanium, and then the enoxacin was covalently bound to the PEG, which was confirmed by X-ray photoelectron spectrometry (XPS). A spread plate assay, confocal laser scanning microscopy (CLSM), and scanning electron microscopy (SEM) were used to characterize the antimicrobial activity. For the in vivo study, Ti implants were inoculated with methicillin-resistant Staphylococcus aureus (MRSA) and implanted into the femoral medullary cavity of rats. The degree of infection was assessed by radiography, micro-computed tomography, and determination of the counts of adherent bacteria 3 weeks after surgery. Our data demonstrate that the enoxacin-modified PEGylated Ti surface effectively prevented bacterial colonization without compromising cell viability, adhesion, or proliferation in vitro. Furthermore, it prevented MRSA infection of the Ti implants in vivo. Taken together, our results demonstrate that the use of enoxacin-modified Ti is a potential approach to the alleviation of infections of Ti implants by multiple bacterial species. PMID:27799220

Effect of postharvest UV-C treatment on the bacterial diversity of Ataulfo mangoes by PCR-DGGE, survival of E. coli and antimicrobial activity

PubMed Central

Fernández-Suárez, Rocío; Ramírez-Villatoro, Guadalupe; Díaz-Ruiz, Gloria; Eslava, Carlos; Calderón, Montserrat; Navarro-Ocaña, Arturo; Trejo-Márquez, Andrea; Wacher, Carmen

2013-01-01

Since Mexico is the second largest exporter of mangoes, its safety assurance is essential. Research in microbial ecology and knowledge of complex interactions among microbes must be better understood to achieve maximal control of pathogens. Therefore, we investigated the effect of UV-C treatments on bacterial diversity of the Ataulfo mangoes surface using PCR-DGGE analysis of variable region V3 of 16S rRNA genes, and the survival of E. coli, by plate counting. The UV-C irradiation reduced the microbial load on the surface of mangoes immediately after treatment and the structure of bacterial communities was modified during storage. We identified the key members of the bacterial communities on the surface of fruits, predominating Enterobacter genus. Genera as Lactococcus and Pantoea were only detected on the surface of non-treated (control) mangoes. This could indicate that these genera were affected by the UV-C treatment. On the other hand, the treatment did not have a significant effect on survival of E. coli. However, genera that have been recognized as antagonists against foodborne pathogens were identified in the bands patterns. Also, phenolic compounds were determined by HPLC and antimicrobial activity was assayed according to the agar diffusion method. The main phenolic compounds were chlorogenic, gallic, and caffeic acids. Mango peel methanol extracts (UV-C treated and control mangoes) showed antimicrobial activity against strains previously isolated from mango, detecting significant differences (P < 0.05) among treated and control mangoes after 4 and 12 days of storage. Ps. fluorescens and Ps. stutszeri were the most sensitive. PMID:23761788

Photon-Counting H33D Detector for Biological Fluorescence Imaging

PubMed Central

Michalet, X.; Siegmund, O.H.W.; Vallerga, J.V.; Jelinsky, P.; Millaud, J.E.; Weiss, S.

2010-01-01

We have developed a photon-counting High-temporal and High-spatial resolution, High-throughput 3-Dimensional detector (H33D) for biological imaging of fluorescent samples. The design is based on a 25 mm diameter S20 photocathode followed by a 3-microchannel plate stack, and a cross delay line anode. We describe the bench performance of the H33D detector, as well as preliminary imaging results obtained with fluorescent beads, quantum dots and live cells and discuss applications of future generation detectors for single-molecule imaging and high-throughput study of biomolecular interactions. PMID:20151021

Cultivable anaerobic and aerobic bacterial communities in the fermentation chambers of Holotrichia parallela (coleoptera: scarabaeidae) larvae.

PubMed

Zhang, Zhen-Yu; Yuan, Yimin; Ali, Muhammad Waqar; Peng, Tao; Peng, Wei; Raza, Muhammad Fahim; Zhao, Yongshun; Zhang, Hongyu

2018-01-01

As important pests, scarab beetle larvae survive on plant biomass and the microbiota of the fermentation chamber play an important role in the digestion of lignocellulose-rich diets. However, the cultivable microbes, especially the anaerobic cultivable microbes, are still largely unknown. Here, both cultivable anaerobic and aerobic bacterial communities associated with the fermentation chamber of Holotrichia parallela larvae were investigated. In total bacteria cells directly enumerated by the 4', 6-diamidino-2-phenylindole (DAPI) staining method, the viable plate counts of cultivable bacteria in the fermentation chamber accounted for 0.92% of proportion. These cultivable bacteria were prone to attach to the fermentation chamber wall (88.41%) compared to the chamber contents. Anaerobic bacteria were dominant in the cultivable bacteria attaching to the fermentation chamber wall (70.20%), while the quantities of anaerobes and aerobes were similar in the chamber contents. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), fingerprinting and sequence analysis of isolated colonies revealed that the cultivable bacteria are affiliated with class γ-Proteobacteria, Bacteroidia, Actinobacteria, Clostridia and β-Proteobacteria. γ-Proteobacteria was the major type of anaerobic cultivable bacteria and even the only one type of aerobic cultivable bacteria. Taken together, our results suggest, for the first time, that anaerobic microbiota are dominant in cultivable bacteria in the special anoxia niche of the fermentation chamber from H. parallela larvae. These bacterial isolates could be a treasure trove for screening lignocellulytic microbes which are essential for the plant biomass digestion of this scarab species.

Cultivable anaerobic and aerobic bacterial communities in the fermentation chambers of Holotrichia parallela (coleoptera: scarabaeidae) larvae

PubMed Central

Ali, Muhammad Waqar; Peng, Tao; Peng, Wei; Raza, Muhammad Fahim; Zhao, Yongshun; Zhang, Hongyu

2018-01-01

As important pests, scarab beetle larvae survive on plant biomass and the microbiota of the fermentation chamber play an important role in the digestion of lignocellulose-rich diets. However, the cultivable microbes, especially the anaerobic cultivable microbes, are still largely unknown. Here, both cultivable anaerobic and aerobic bacterial communities associated with the fermentation chamber of Holotrichia parallela larvae were investigated. In total bacteria cells directly enumerated by the 4’, 6-diamidino-2-phenylindole (DAPI) staining method, the viable plate counts of cultivable bacteria in the fermentation chamber accounted for 0.92% of proportion. These cultivable bacteria were prone to attach to the fermentation chamber wall (88.41%) compared to the chamber contents. Anaerobic bacteria were dominant in the cultivable bacteria attaching to the fermentation chamber wall (70.20%), while the quantities of anaerobes and aerobes were similar in the chamber contents. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), fingerprinting and sequence analysis of isolated colonies revealed that the cultivable bacteria are affiliated with class γ-Proteobacteria, Bacteroidia, Actinobacteria, Clostridia and β-Proteobacteria. γ-Proteobacteria was the major type of anaerobic cultivable bacteria and even the only one type of aerobic cultivable bacteria. Taken together, our results suggest, for the first time, that anaerobic microbiota are dominant in cultivable bacteria in the special anoxia niche of the fermentation chamber from H. parallela larvae. These bacterial isolates could be a treasure trove for screening lignocellulytic microbes which are essential for the plant biomass digestion of this scarab species. PMID:29304141

USE OF SCORE AND CEREBROSPINAL FLUID LACTATE DOSAGE IN DIFFERENTIAL DIAGNOSIS OF BACTERIAL AND ASEPTIC MENINGITIS.

PubMed

Pires, Frederico Ribeiro; Franco, Andréia Christine Bonotto Farias; Gilio, Alfredo Elias; Troster, Eduardo Juan

2017-01-01

To evaluate Bacterial Meningitis Score (BMS) on its own and in association with Cerebrospinal Fluid (CSF) lactate dosage in order to distinguish bacterial from aseptic meningitis. Children diagnosed with meningitis at a tertiary hospital between January/2011 and December/2014 were selected. All data were obtained upon admission. BMS was applied and included: CSF Gram staining (2 points); CSF neutrophil count ≥1,000 cells/mm3 (1 point); CSF protein ≥80 mg/dL (1 point); peripheral blood neutrophil count ≥10,000 cells/mm3 (1 point) and seizures upon/before arrival (1 point). Cutoff value for CSF lactate was ≥30 mg/dL. Sensitivity, specificity and negative predictive value of several BMS cutoffs and BMS associated with high CSF lactate were evaluated for prediction of bacterial meningitis. Among 439 eligible patients, 94 did not have all data available to complete the score, and 345 patients were included: 7 in bacterial meningitis group and 338 in aseptic meningitis group. As predictive factors of bacterial meningitis, BMS ≥1 had 100% sensitivity (95%CI 47.3-100), 64.2% specificity (58.8-100) and 100% negative predictive value (97.5-100); BMS ≥2 or BMS ≥1 associated with high CSF lactate also showed 100% sensitivity (47.3-100); but 98.5% specificity (96.6-99.5) and 100% negative predictive value (98.3-100). 2 point BMS in association with CSF lactate dosage had the same sensitivity and negative predictive value, with increased specificity for diagnosis of bacterial meningitis when compared with 1-point BMS.

Bacterial quality and safety of packaged fresh leafy vegetables at the retail level in Finland.

PubMed

Nousiainen, L-L; Joutsen, S; Lunden, J; Hänninen, M-L; Fredriksson-Ahomaa, M

2016-09-02

Consumption of packaged fresh leafy vegetables, which are convenient ready-to-eat products, has increased during the last decade. The number of foodborne outbreaks associated with these products has concurrently increased. In our study, (1) label information, (2) O2/CO2 composition, (3) bacterial quality and (4) safety of 100 fresh leafy vegetables at the retail level were studied in Finland during 2013. Bacterial quality was studied using aerobic bacteria (AB) and coliform bacteria (CB) counts, and searching for the presence of Escherichia coli, Listeria and Yersinia. The safety was studied by the presence of Salmonella, ail-positive Yersinia, stx-positive E. coli (STEC) and Listeria monocytogenes using PCR and culturing. Important label information was unavailable on several packages originating from different companies. The packaging date was missing on all packages and the date of durability on 83% of the packages. Storage temperature was declared on 62% of the packages and 73% of the packages contained information about prewashing. The batch/lot number was missing on 29% of the packages. Very low oxygen (O2) (<1%) and elevated carbon dioxide (CO2) (2-22%) concentrations were measured in all packages labelled to contain a protective atmosphere. O2 and CO2 concentrations varied widely in the rest of the packages. AB and CB counts were high in the leafy vegetable samples varying between 6.2 and 10.6 and 4.2-8.3logcfu/g, respectively. In most of the samples, the AB and CB counts exceeded 10(8) and 10(6)cfu/g, respectively. A positive correlation was observed between the AB and CB counts. E. coli was isolated from 15% of the samples and Yersinia from 33%. L. monocytogenes was isolated from two samples and ail-positive Y. enterocolitica in one. Using PCR, STEC was detected in seven samples, and Salmonella and ail-positive Y. enterocolitica in two samples each. The AB and CB mean values of products originating from different companies varied widely. High AB and CB

Microbiological and molecular characterization of commercially available probiotics containing Bacillus clausii from India and Pakistan.

PubMed

Patrone, Vania; Molinari, Paola; Morelli, Lorenzo

2016-11-21

Probiotics are actively used for treatment of diarrhoea, respiratory infections, and prevention of infectious gastrointestinal diseases. The efficacy of probiotics is due to strain-specific features and the number of viable cells; however, several reports of deviations from the label in the actual content of strains in probiotic products are a matter of concern. Most of the available data on quality focuses on probiotic products containing lactobacilli and/or bifidobacteria, while very few data are available on spore-forming probiotics. The present study evaluates the label claims for spore count and species identification in five commercial probiotic products marketed in India and Pakistan that claim to contain Bacillus clausii: Tufpro, Ecogro, Enterogermina, Entromax, and Ospor. Bacterial enumeration from three batches was done by microbiological plating methods by two independent operators. Species identification was done using PCR amplification and sequence analysis of the 16S rRNA gene, and determination of the total amount of species present in the products was done using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analysis followed by DNA sequencing of the excised bands. Plate count methods demonstrated poor correlations between quantitative label indications and bacteria recovered from plates for Tufpro, Ecogro, and Ospor. The 16S rRNA analysis performed on bacteria isolated from plate counts showed that only Enterogermina and Ospor contained homogenous B. clausii. PCR-DGGE analysis revealed that only Enterogermina had a homogenous B. clausii population while other products had mixed bacterial populations. In conclusion, the current analysis clearly demonstrates that of the five analysed commercial probiotics, only Enterogermina followed the label claims. Copyright © 2016 Elsevier B.V. All rights reserved.

Diagnostic value of lactate, procalcitonin, ferritin, serum-C-reactive protein, and other biomarkers in bacterial and viral meningitis

PubMed Central

Sanaei Dashti, Anahita; Alizadeh, Shekoofan; Karimi, Abdullah; Khalifeh, Masoomeh; Shoja, Seyed Abdolmajid

2017-01-01

Abstract There are many difficulties distinguishing bacterial from viral meningitis that could be reasonably solved using biomarkers. The aim of this study was to evaluate lactate, procalcitonin (PCT), ferritin, serum-CRP (C-reactive protein), and other known biomarkers in differentiating bacterial meningitis from viral meningitis in children. All children aged 28 days to 14 years with suspected meningitis who were admitted to Mofid Children's Hospital, Tehran, between October 2012 and November 2013, were enrolled in this prospective cross-sectional study. Children were divided into 2 groups of bacterial and viral meningitis, based on the results of cerebrospinal fluid (CSF) culture, polymerase chain reaction, and cytochemical profile. Diagnostic values of CSF parameters (ferritin, PCT, absolute neutrophil count [ANC], white blood cell count, and lactate) and serum parameters (PCT, ferritin, CRP, and erythrocyte sedimentation rate [ESR]) were evaluated. Among 50 patients with meningitis, 12 were diagnosed with bacterial meningitis. Concentrations of all markers were significantly different between bacterial and viral meningitis, except for serum (P = .389) and CSF (P = .136) PCT. The best rates of area under the receiver operating characteristic (ROC) curve (AUC) were achieved by lactate (AUC = 0.923) and serum-CRP (AUC = 0.889). The best negative predictive values (NPV) for bacterial meningitis were attained by ANC (100%) and lactate (97.1%). The results of our study suggest that ferritin and PCT are not strong predictive biomarkers. A combination of low CSF lactate, ANC, ESR, and serum-CRP could reasonably rule out the bacterial meningitis. PMID:28858084

Bacterial communities in ancient permafrost profiles of Svalbard, Arctic.

PubMed

Singh, Purnima; Singh, Shiv M; Singh, Ram N; Naik, Simantini; Roy, Utpal; Srivastava, Alok; Bölter, Manfred

2017-12-01

Permafrost soils are unique habitats in polar environment and are of great ecological relevance. The present study focuses on the characterization of bacterial communities from permafrost profiles of Svalbard, Arctic. Counts of culturable bacteria range from 1.50 × 10 3 to 2.22 × 10 5 CFU g -1 , total bacterial numbers range from 1.14 × 10 5 to 5.52 × 10 5 cells g -1 soil. Bacterial isolates are identified through 16S rRNA gene sequencing. Arthrobacter and Pseudomonas are the most dominant genera, and A. sulfonivorans, A. bergeri, P. mandelii, and P. jessenii as the dominant species. Other species belong to genera Acinetobacter, Bacillus, Enterobacter, Nesterenkonia, Psychrobacter, Rhizobium, Rhodococcus, Sphingobacterium, Sphingopyxis, Stenotrophomonas, and Virgibacillus. To the best of our knowledge, genera Acinetobacter, Enterobacter, Nesterenkonia, Psychrobacter, Rhizobium, Sphingobacterium, Sphingopyxis, Stenotrophomonas, and Virgibacillus are the first northernmost records from Arctic permafrost. The present study fills the knowledge gap of culturable bacterial communities and their chronological characterization from permafrost soils of Ny-Ålesund (79°N), Arctic. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A Treatise on Equivalent-Plate Stiffnesses for Stiffened Laminated-Composite Plates and Plate-Like Lattices

NASA Technical Reports Server (NTRS)

Nemeth, Michael P.

2011-01-01

A survey of studies conducted since 1914 on the use of equivalent-plate stiffnesses in modeling the overall, stiffness-critical response of stiffened plates and shells is presented. Two detailed, comprehensive derivations of first-approximation equivalent-plate stiffnesses are also presented that are based on the Reissner-Mindlin-type, first-order transverse-shear deformation theory for anisotropic plates. Equivalent-plate stiffness expressions, and a corresponding symbolic manipulation computer program, are also presented for several different stiffener configurations. These expressions are very general and exhibit the full range of anisotropies permitted by the Reissner-Mindlin-type, first-order transverse-shear deformation theory for anisotropic plates. The expressions presented in the present study were also compared with available, previously published results. For the most part, the previously published results are for special cases of the general expressions presented herein and are almost in complete agreement. Analysis is also presented that extends the use of the equivalent-plate stiffness expressions to sandwich plates.

Quick counting method for estimating the number of viable microbes on food and food processing equipment.

PubMed

Winter, F H; York, G K; el-Nakhal, H

1971-07-01

A rapid method for estimating the extent of microbial contamination on food and on food processing equipment is described. Microbial cells are rinsed from food or swab samples with sterile diluent and concentrated on the surface of membrane filters. The filters are incubated on a suitable bacteriological medium for 4 hr at 30 C, heated at 105 C for 5 min, and stained. The membranes are then dried at 60 C for 15 min, rendered transparent with immersion oil, and examined microscopically. Data obtained by the rapid method were compared with counts of the same samples determined by the standard plate count method. Over 60 comparisons resulted in a correlation coefficient of 0.906. Because the rapid technique can provide reliable microbiological count information in extremely short times, it can be a most useful tool in the routine evaluation of microbial contamination of food processing facilities and for some foods.

Effects of lead and zinc mining contamination on bacterial community diversity and enzyme activities of vicinal cropland.

PubMed

Qu, Juanjuan; Ren, Guangming; Chen, Bao; Fan, Jinghua; E, Yong

2011-11-01

In the process of mining activity, many kinds of heavy metals enter into soils with dust, causing serious contamination to the environment. In this study, six soils were sampled from cropland at different distances from a lead/zinc mine in Heilongjiang Province, China. The total contents of lead and zinc in the vicinal cropland exceeded the third level of environmental quality standard for soil in China, which indicated that soils in this area were moderately contaminated. Bacterial community diversity and population were greatly decreased when the concentrations of lead and zinc were beyond 1,500 and 995 mg kg(-1), respectively, as analyzed by plate counting and polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). The bands of DGGE patterns varied with the degree of contamination. The activities of soil urease, phosphatase, and dehydrogenase were negatively correlated with the concentrations of lead and zinc. The highest inhibitory effect of heavy metals on soil enzyme activities was observed in urease. It was noted that PCR-DGGE patterns combined with soil enzyme activity analysis can be indices for the soil quality assessment by heavy metal contamination.

Choral Counting

ERIC Educational Resources Information Center

Turrou, Angela Chan; Franke, Megan L.; Johnson, Nicholas

2017-01-01

The students in Ms. Moscoso's second-grade class gather on the rug after recess, ready for one of their favorite math warm-ups: Choral Counting. Counting is an important part of doing mathematics throughout the school; students count collections (Schwerdtfeger and Chan 2007) and solve problems using a variety of strategies, many of which are…

Influence of protein deposition on bacterial adhesion to contact lenses.

PubMed

Subbaraman, Lakshman N; Borazjani, Roya; Zhu, Hua; Zhao, Zhenjun; Jones, Lyndon; Willcox, Mark D P

2011-08-01

The aim of the study is to determine the adhesion of Gram positive and Gram negative bacteria onto conventional hydrogel (CH) and silicone hydrogel (SH) contact lens materials with and without lysozyme, lactoferrin, and albumin coating. Four lens types (three SH-balafilcon A, lotrafilcon B, and senofilcon A; one CH-etafilcon A) were coated with lysozyme, lactoferrin, or albumin (uncoated lenses acted as controls) and then incubated in Staphylococcus aureus (Saur 31) or either of two strains of Pseudomonas aeruginosa (Paer 6294 and 6206) for 24 h at 37 °C. The total counts of the adhered bacteria were determined using the H-thymidine method and viable counts by counting the number of colony-forming units on agar media. All three strains adhered significantly lower to uncoated etafilcon A lenses compared with uncoated SH lenses (p < 0.05). Lysozyme coating on all four lens types increased binding (total and viable counts) of Saur 31 (p < 0.05). However, lysozyme coating did not influence P. aeruginosa adhesion (p > 0.05). Lactoferrin coating on lenses increased binding (total and viable counts) of Saur 31 (p < 0.05). Lactoferrin-coated lenses showed significantly higher total counts (p < 0.05) but significantly lower viable counts (p < 0.05) of adhered P. aeruginosa strains. There was a significant difference between the total and viable counts (p < 0.05) that were bound to lactoferrin-coated lenses. Albumin coating of lenses increased binding (total and viable counts) of all three strains (p < 0.05). Lysozyme deposited on contact lenses does not possess antibacterial activity against certain bacterial strains, whereas lactoferrin possess an antibacterial effect against strains of P. aeruginosa.

Cold plate

DOEpatents

Marroquin, Christopher M.; O'Connell, Kevin M.; Schultz, Mark D.; Tian, Shurong

2018-02-13

A cold plate, an electronic assembly including a cold plate, and a method for forming a cold plate are provided. The cold plate includes an interface plate and an opposing plate that form a plenum. The cold plate includes a plurality of active areas arranged for alignment over respective heat generating portions of an electronic assembly, and non-active areas between the active areas. A cooling fluid flows through the plenum. The plenum, at the non-active areas, has a reduced width and/or reduced height relative to the plenum at the active areas. The reduced width and/or height of the plenum, and exterior dimensions of cold plate, at the non-active areas allow the non-active areas to flex to accommodate surface variations of the electronics assembly. The reduced width and/or height non-active areas can be specifically shaped to fit between physical features of the electronics assembly.

Clinical Value of Assessing Cytokine Levels for the Differential Diagnosis of Bacterial Meningitis in a Pediatric Population

PubMed Central

Ye, Qing; Shao, Wen-Xia; Shang, Shi-Qiang; Shen, Hong-Qiang; Chen, Xue-Jun; Tang, Yong-Min; Yu, Yong-Lin; Mao, Jian-Hua

2016-01-01

Abstract We performed a prospective observational study to evaluate the utility of measuring inflammatory cytokine levels to discriminate bacterial meningitis from similar common pediatric diseases. Inflammatory cytokine levels and other cerebrospinal fluid (CSF) physicochemical indicators were evaluated in 140 patients who were diagnosed with bacterial meningitis via microbiological culture or PCR assay. The CSF concentrations of interleukin (IL)-6 and IL-10, CSF/blood IL-6 and IL-10 ratios, CSF white blood cell count, and CSF micro total protein were significantly elevated in bacterial meningitis patients compared with healthy children or patients with viral encephalitis, epilepsy, or febrile convulsions (P < 0.001). The area under the curve values for CSF concentrations of IL-6 and IL-10, CSF/blood IL-6 and IL-10 ratios, CSF white blood cell count, and CSF micro total protein to identify bacterial meningitis episodes by receiver-operating characteristic analysis were 0.988, 0.949, 0.995, 0.924, 0.945, and 0.928, respectively. The area under the curve for the combination of CSF IL-6 and CSF/blood IL-6 ratio was larger than that for either parameter alone, and the combination exhibited enhanced specificity and positive predictive value. After effective meningitis treatment, CSF IL-6 levels dropped significantly. These results suggest that CSF IL-6 and CSF/blood IL-6 ratio are good biomarkers in discriminating bacterial meningitis. Evaluating CSF IL-6 and CSF/blood IL-6 ratio in combination can improve diagnostic efficiency. Additionally, CSF IL-6 levels can be used to monitor the effects of bacterial meningitis treatment. PMID:27043692

Clinical Value of Assessing Cytokine Levels for the Differential Diagnosis of Bacterial Meningitis in a Pediatric Population.

PubMed

Ye, Qing; Shao, Wen-Xia; Shang, Shi-Qiang; Shen, Hong-Qiang; Chen, Xue-Jun; Tang, Yong-Min; Yu, Yong-Lin; Mao, Jian-Hua

2016-03-01

We performed a prospective observational study to evaluate the utility of measuring inflammatory cytokine levels to discriminate bacterial meningitis from similar common pediatric diseases. Inflammatory cytokine levels and other cerebrospinal fluid (CSF) physicochemical indicators were evaluated in 140 patients who were diagnosed with bacterial meningitis via microbiological culture or PCR assay. The CSF concentrations of interleukin (IL)-6 and IL-10, CSF/blood IL-6 and IL-10 ratios, CSF white blood cell count, and CSF micro total protein were significantly elevated in bacterial meningitis patients compared with healthy children or patients with viral encephalitis, epilepsy, or febrile convulsions (P < 0.001). The area under the curve values for CSF concentrations of IL-6 and IL-10, CSF/blood IL-6 and IL-10 ratios, CSF white blood cell count, and CSF micro total protein to identify bacterial meningitis episodes by receiver-operating characteristic analysis were 0.988, 0.949, 0.995, 0.924, 0.945, and 0.928, respectively. The area under the curve for the combination of CSF IL-6 and CSF/blood IL-6 ratio was larger than that for either parameter alone, and the combination exhibited enhanced specificity and positive predictive value. After effective meningitis treatment, CSF IL-6 levels dropped significantly. These results suggest that CSF IL-6 and CSF/blood IL-6 ratio are good biomarkers in discriminating bacterial meningitis. Evaluating CSF IL-6 and CSF/blood IL-6 ratio in combination can improve diagnostic efficiency. Additionally, CSF IL-6 levels can be used to monitor the effects of bacterial meningitis treatment.

Cerebrospinal fluid ferritin in children with viral and bacterial meningitis.

PubMed

Rezaei, M; Mamishi, S; Mahmoudi, S; Pourakbari, B; Khotaei, G; Daneshjou, K; Hashemi, N

2013-01-01

Despite the fact that the prognosis of bacterial meningitis has been improved by the influence of antibiotics, this disease is still one of the significant causes of morbidity and mortality in children. Rapid differentiation between bacterial and aseptic meningitis, and the need for immediate antibiotic treatment in the former, is crucial in the prognosis of these patients. Ferritin is one of the most sensitive biochemical markers investigated in cerebrospinal fluid (CSF) for the early diagnosis of bacterial meningitis. The present study aims to evaluate the diagnostic capability of CSF ferritin in differentiating bacterial and viral meningitis in the paediatric setting. A cross-sectional study was carried out in the referral Children's Medical Center Hospital, Tehran, during 2008 and 2009. According to the inclusion criteria, CSF samples from 42 patients with suspected meningitis were obtained and divided into two meningitis groups, bacterial (n = 18) and viral (n = 24). Ferritin and other routine determinants (i.e., leucocytes, protein and glucose) were compared between the two groups. Ferritin concentration in the bacterial meningitis group was 106.39 +/- 86.96 ng/dL, which was considerably higher than in the viral meningitis group (10.17 +/- 14.09, P < 0.001). Mean CSF protein concentration and cell count were significantly higher in the bacterial meningitis group and showed a positive correlation with CSF ferritin. In conclusion, this study suggests that CSF ferritin concentration is an accurate test for the early differentiation of bacterial and aseptic meningitis; however, further investigation on a larger cohort of patients is required to confirm this finding.

Comparison of the cytotoxic effect of polystyrene latex nanoparticles on planktonic cells and bacterial biofilms

NASA Astrophysics Data System (ADS)

Nomura, Toshiyuki; Fujisawa, Eri; Itoh, Shikibu; Konishi, Yasuhiro

2016-06-01

The cytotoxic effect of positively charged polystyrene latex nanoparticles (PSL NPs) was compared between planktonic bacterial cells and bacterial biofilms using confocal laser scanning microscopy, atomic force microscopy, and a colony counting method. Pseudomonas fluorescens, which is commonly used in biofilm studies, was employed as the model bacteria. We found that the negatively charged bacterial surface of the planktonic cells was almost completely covered with positively charged PSL NPs, leading to cell death, as indicated by the NP concentration being greater than that required to achieve single layer coverage. In addition, the relationship between surface coverage and cell viability of P. fluorescens cells correlated well with the findings in other bacterial cells ( Escherichia coli and Lactococcus lactis). However, most of the bacterial cells that formed the biofilm were viable despite the positively charged PSL NPs being highly toxic to planktonic bacterial cells. This indicated that bacterial cells embedded in the biofilm were protected by self-produced extracellular polymeric substances (EPS) that provide resistance to antibacterial agents. In conclusion, mature biofilms covered with EPS exhibit resistance to NP toxicity as well as antibacterial agents.

Role of neutrophil to lymphocyte and monocyte to lymphocyte ratios in the diagnosis of bacterial infection in patients with fever.

PubMed

Naess, Are; Nilssen, Siri Saervold; Mo, Reidun; Eide, Geir Egil; Sjursen, Haakon

2017-06-01

To study the role of the neutrophil:lymphocyte ratio (NLR) and monocyte:lymphocyte ratio (MLR) in discriminating between different patient groups hospitalized for fever due to infection and those without infection. For 299 patients admitted to hospital for fever with unknown cause, a number of characteristics including NLR and MLR were recorded. These characteristics were used in a multiple multinomial regression analysis to estimate the probability of a final diagnostic group of bacterial, viral, clinically confirmed, or no infection. Both NLR and MLR significantly predicted final diagnostic group. Being highly correlated, however, both variables could not be retained in the same model. Both variables also interacted significantly with duration of fever. Generally, higher values of NLR and MLR indicated larger probabilities for bacterial infection and low probabilities for viral infection. Patients with septicemia had significantly higher NLR compared to patients with other bacterial infections with fever for less than one week. White blood cell counts, neutrophil counts, and C-reactive proteins did not differ significantly between septicemia and the other bacterial infection groups. NLR is a more useful diagnostic tool to identify patients with septicemia than other more commonly used diagnostic blood tests. NLR and MLR may be useful in the diagnosis of bacterial infection among patients hospitalized for fever.

Mutacins and bacteriocins like genes in Streptococcus mutans isolated from participants with high, moderate, and low salivary count.

PubMed

Soto, Carolina; Padilla, Carlos; Lobos, Olga

2017-02-01

To detect S. mutans producers of mutacins and bacteriocins like substances (BLIS) from saliva of participants with low, moderate, and high salivary counts. 123 strains of S. mutans were obtained from participants with low, moderate, and high salivary counts (age 18 and 20 years old) and their antibacterial capacity analyzed. By using PCR amplification, the expression levels of mutacins and BLIS genes were studied (expressed in arbitrary units/ml) in all three levels. S. mutans strains from participants with low salivary counts show high production of mutacins (63%). In contrast, participants with moderate and high salivary counts depict relatively low levels of mutacins (22 and 15%, respectively). Moreover, participants with low salivary counts showed high expression levels of genes encoding mutacins, a result that correlates with the strong antimicrobial activity of the group. Participants with moderate and high salivary counts however depict low expression levels of mutacin related genes, and little antimicrobial activity. No BLIS were detected in any of the groups studied. S. mutans isolated from the saliva of participants with low bacterial counts have significant antibacterial capacity compared to that of participants with moderate and high salivary counts. The superior lethality of S. mutans in participants with low salivary counts is likely due to the augmented expression of mutacin- related genes. Copyright © 2016 Elsevier Ltd. All rights reserved.

CSF lactate for accurate diagnosis of community-acquired bacterial meningitis.

PubMed

Giulieri, S; Chapuis-Taillard, C; Jaton, K; Cometta, A; Chuard, C; Hugli, O; Du Pasquier, R; Bille, J; Meylan, P; Manuel, O; Marchetti, O

2015-10-01

CSF lactate measurement is recommended when nosocomial meningitis is suspected, but its value in community-acquired bacterial meningitis is controversial. We evaluated the diagnostic performance of lactate and other CSF parameters in a prospective cohort of adult patients with acute meningitis. Diagnostic accuracy of lactate and other CSF parameters in patients with microbiologically documented episodes was assessed by receiver operating characteristic (ROC) curves. The cut-offs with the best diagnostic performance were determined. Forty-five of 61 patients (74%) had a documented bacterial (n = 18; S. pneumoniae, 11; N. meningitidis, 5; other, 2) or viral (n = 27 enterovirus, 21; VZV, 3; other, 3) etiology. CSF parameters were significantly different in bacterial vs. viral meningitis, respectively (p < 0.001 for all comparisons): white cell count (median 1333 vs. 143/mm(3)), proteins (median 4115 vs. 829 mg/l), CSF/blood glucose ratio (median 0.1 vs. 0.52), lactate (median 13 vs. 2.3 mmol/l). ROC curve analysis showed that CSF lactate had the highest accuracy for discriminating bacterial from viral meningitis, with a cutoff set at 3.5 mmol/l providing 100% sensitivity, specificity, PPV, NPV, and efficiency. CSF lactate had the best accuracy for discriminating bacterial from viral meningitis and should be included in the initial diagnostic workup of this condition.

USE OF SCORE AND CEREBROSPINAL FLUID LACTATE DOSAGE IN DIFFERENTIAL DIAGNOSIS OF BACTERIAL AND ASEPTIC MENINGITIS

PubMed Central

Pires, Frederico Ribeiro; Franco, Andréia Christine Bonotto Farias; Gilio, Alfredo Elias; Troster, Eduardo Juan

2017-01-01

ABSTRACT Objective: To evaluate Bacterial Meningitis Score (BMS) on its own and in association with Cerebrospinal Fluid (CSF) lactate dosage in order to distinguish bacterial from aseptic meningitis. Methods: Children diagnosed with meningitis at a tertiary hospital between January/2011 and December/2014 were selected. All data were obtained upon admission. BMS was applied and included: CSF Gram staining (2 points); CSF neutrophil count ≥1,000 cells/mm3 (1 point); CSF protein ≥80 mg/dL (1 point); peripheral blood neutrophil count ≥10,000 cells/mm3 (1 point) and seizures upon/before arrival (1 point). Cutoff value for CSF lactate was ≥30 mg/dL. Sensitivity, specificity and negative predictive value of several BMS cutoffs and BMS associated with high CSF lactate were evaluated for prediction of bacterial meningitis. Results: Among 439 eligible patients, 94 did not have all data available to complete the score, and 345 patients were included: 7 in bacterial meningitis group and 338 in aseptic meningitis group. As predictive factors of bacterial meningitis, BMS ≥1 had 100% sensitivity (95%CI 47.3-100), 64.2% specificity (58.8-100) and 100% negative predictive value (97.5-100); BMS ≥2 or BMS ≥1 associated with high CSF lactate also showed 100% sensitivity (47.3-100); but 98.5% specificity (96.6-99.5) and 100% negative predictive value (98.3-100). Conclusions: 2 point BMS in association with CSF lactate dosage had the same sensitivity and negative predictive value, with increased specificity for diagnosis of bacterial meningitis when compared with 1-point BMS. PMID:29185620

A novel lignin degradation bacterial consortium for efficient pulping.

PubMed

Wang, Yanxia; Liu, Quan; Yan, Lei; Gao, Yamei; Wang, Yanjie; Wang, Weidong

2013-07-01

A lignin degradation bacterial consortium named LDC was screened from the sludge of a reeds pond by a restricted subculture. It could break down 60.9% lignin in reeds at 30°C under conditions of static culture within 15 days. In order to analyze the diversity of LDC, plate isolation, 16S rDNA clone library and ARDRA (Amplified Ribosomal DNA Restriction Analysis) were performed. Six bacterial strains were isolated from LDC and eighteen DNA phylotypes were identified from 230 bacterial analyzed clones. They were classified into Clostridiales(9.1%), Geovibrio thiophilus (5.1%), Desulfomicrobium (10.9%), Pseudomonas sp. (25.2%), Azoarcus sp. (5.1%), Thauera (5.1%), Paenibacillus sp. (5.1%), Cohnella sp. (2.2%), Acinetobacter sp. (3.1%), Microbacterium (7.8%), and uncultured bacterium (21.3%). In addition, physical characteristics of paper hand-sheets between biological pretreatment and chemical pretreatment were compared. The results showed that LDC had the capability of lignin degradation and was efficient for pulping, which would provide a new choice for biopulping. Copyright © 2013 Elsevier Ltd. All rights reserved.

Health Threats from Contamination of Spices Commercialized in Romania: Risks of Fungal and Bacterial Infections.

PubMed

Man, Adrian; Mare, Anca; Toma, Felicia; Curticăpean, Augustin; Santacroce, Luigi

2016-01-01

The study of fungal contamination in food and mycotoxicoses is a priority today, both internationally and nationally. The purpose of this study is to have a general view over the quality of the most common spices that are sold in Romanian markets, by assessing the degree of fungal, bacterial and mycotoxin contamination in pepper and chili powders. We tested four types of spices: white pepper, black pepper, sweet and hot chili powders from 12 different distributing companies, summing a total of 35 sample types. The fungal and bacterial load was assessed by Standard Plate Count, while the mycotoxin content by High-performance liquid chromatography. Environmental conditions (humidity, pH) and the selling price for each product were also followed. Fungi were observed in 72.7% of black pepper samples, 33.3% in white pepper, 30% in sweet chili and 25% in hot chili products. The most common isolated fungus was Aspergillus spp., while Rhizopus, Mucor, Fusarium, Penicillium, Absidia species were found, in smaller percentage. Four producers (44.4%) presented fungal contamination of over 10^3 CFU/g and two producers (22.2%) presented no fungal contamination in their products. Bacterial contamination was found in 85.7% of the tested products, consisting mostly in Bacillus spp. Aflatoxin B1 was present in all the tested products, mostly in black pepper (mean value 126.3 ng/g); Ochratoxin A was present in sweet chili (mean value 328 ng/g) and Zearalenone in hot chili (mean value 604 ng/g) and sweet chili (mean value 382 ng/g). All spices presented either fungal contamination, mycotoxin contamination, or both. The high humidity and the high pH of spices represent favorable conditions for fungal growth. The selling price was partly related to the physic-chemical conditions and microbiological quality of the spices. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Fluorometry as a bacterial source tracking tool in coastal watersheds, Trinidad, CA

Treesearch

Trever Parker; Andrew Stubblefield

2012-01-01

Bacterial counts have long been used as indicators of water pollution that may affect public health. By themselves, bacteria are indicators only and can not be used to identify the source of the pollutant for remediation efforts. Methods of microbial source tracking are generally time consuming, labor intensive and expensive. As an alternative, a fluorometer can be...

Bacterial contamination of ultrasound probes in different radiological institutions before and after specific hygiene training: do we have a general hygienical problem?

PubMed

Sartoretti, Thomas; Sartoretti, Elisabeth; Bucher, Candid; Doert, Aleksis; Binkert, Christoph; Hergan, Klaus; Meissnitzer, Matthias; Froehlich, Johannes; Kolokythas, Orpheus; Matoori, Simon; Orasch, Christina; Kos, Sebastian; Sartoretti-Schefer, Sabine; Gutzeit, Andreas

2017-10-01

Aim was to investigate hygienic conditions of ultrasound probes before and after hygiene training in radiology institutions in comparison to bacterial contamination in public places. In three radiology departments, bacterial contamination was evaluated using baseline agar plates for cultures taken from 36 ultrasound probes. Afterwards teams were trained by a hygiene service centre and 36 ultrasound probes were routinely disinfected with regular disinfecting wipes and then evaluated. In comparison, bacterial contamination in public places (bus poles, n = 11; toilet seats, n = 10) were analysed. Plates were routinely incubated and the number of colony forming units (CFU) analysed. Cultures taken from the probes showed a median of 53 CFU before and 0 CFU after training (p < 0.001). Cultures taken from public places showed a median of 4 CFU from toilets and 28 from bus poles and had lower bacterial load in comparison to ultrasound probes before training (p = 0.055, toilets; p = 0.772, bus poles), without statistical significance. Bacterial contamination of ultrasound probes prior to hygiene training proved to be high and showed higher bacterial load than toilets seats or bus poles. Radiologists should be aware that the lack of hygiene in the field of ultrasound diagnostics puts patients at risk of healthcare-associated infections. • Hospital-associated infections are a problem for patient care. • Hygiene training of staff prevents bacterial contamination of ultrasound probes. • Disinfection of ultrasound probes is an easy method to protect patients.

Community-acquired bacterial meningitis in adults with cancer or a history of cancer.

PubMed

Costerus, Joost M; Brouwer, Matthijs C; van der Ende, Arie; van de Beek, Diederik

2016-03-01

To study the incidence, clinical presentation, causative bacteria, and outcome of community-acquired bacterial meningitis in adults with cancer. We evaluated incidence and characteristics of patients with cancer included in a nationwide prospective cohort study of adults with community-acquired meningitis performed in the Netherlands from March 1, 2006, to September 31, 2014. All patients underwent a neurologic examination at hospital discharge, and outcome was graded using the Glasgow Outcome Scale. Active cancer was identified in 68 of 1,351 episodes (5%) and a history of cancer in 87 (6%). The annual incidence of community-acquired bacterial meningitis was 2.71-fold (95% confidence interval [CI] 1.68-4.36, p < 0.001) increased for patients with cancer compared to patients without cancer in 2010, and 3.52-fold (95% CI 2.16-5.73, p < 0.001) in 2013. The clinical presentation of bacterial meningitis in patients with cancer compared to patients without cancer was similar. Patients with active cancer presented with lower leukocyte count in blood (12.1 × 10(9) cells/L vs 17.3 × 10(9) cells/L, p < 0.001) and CSF (670 cells/mm(3) vs 2,567 cells/mm(3), p < 0.001) and were more likely to be infected with Listeria monocytogenes (21% vs 5%, p < 0.001) than patients without cancer. Active cancer was identified as an independent risk factor for unfavorable outcome in bacterial meningitis (odds ratio 1.85, 95% CI 1.09-3.13). One of 8 patients with community-bacterial meningitis was identified to have a history of cancer and cancer was considered active in half of these patients. Patients with active cancer present with lower CSF leukocyte counts, are more likely to be infected with L monocytogenes, and are at high risk of unfavorable outcome. © 2016 American Academy of Neurology.

Surface topography of composite restorative materials following ultrasonic scaling and its Impact on bacterial plaque accumulation. An in-vitro SEM study.

PubMed

Hossam, A Eid; Rafi, A Togoo; Ahmed, A Saleh; Sumanth, Phani Cr

2013-06-01

This is an in vitro study to investigate the effects of ultrasonic scaling on the surface roughness and quantitative bacterial count on four different types of commonly used composite restorative materials for class V cavities. Nanofilled, hybrid, silorane and flowable composites were tested. Forty extracted teeth served as specimen and were divided into 4 groups of 10 specimens, with each group receiving a different treatment and were examined by a Field emission scanning electron microscope. Bacterial suspension was then added to the pellicle-coated specimens, and then bacterial adhesion was analyzed by using image analyzing program. Flowable and silorane-based composites showed considerably smoother surfaces and lesser bacterial count in comparison to other types, proving that bacterial adhesion is directly proportional to surface roughness. The use of ultrasonic scalers affects the surfaces of composite restorative materials. Routine periodontal scaling should be carried out very carefully, and polishing of the scaled surfaces may overcome the alterations in roughness, thus preventing secondary caries, surface staining, plaque accumulation and subsequent periodontal inflammation. How to cite this article: Eid H A, Togoo R A, Saleh A A, Sumanth C R. Surface Topography of Composite Restorative Materials following Ultrasonic Scaling and its Impact on Bacterial Plaque Accumulation. An In-Vitro SEM Study. J Int Oral Health 2013; 5(3):13-19.

Surface topography of composite restorative materials following ultrasonic scaling and its Impact on bacterial plaque accumulation. An in-vitro SEM study

PubMed Central

Hossam, A. Eid; Rafi, A. Togoo; Ahmed, A Saleh; Sumanth, Phani CR

2013-01-01

Background: This is an in vitro study to investigate the effects of ultrasonic scaling on the surface roughness and quantitative bacterial count on four different types of commonly used composite restorative materials for class V cavities. Materials & Methods: Nanofilled, hybrid, silorane and flowable composites were tested. Forty extracted teeth served as specimen and were divided into 4 groups of 10 specimens, with each group receiving a different treatment and were examined by a Field emission scanning electron microscope. Bacterial suspension was then added to the pellicle-coated specimens, and then bacterial adhesion was analyzed by using image analyzing program. Results: Flowable and silorane-based composites showed considerably smoother surfaces and lesser bacterial count in comparison to other types, proving that bacterial adhesion is directly proportional to surface roughness. Conclusion: The use of ultrasonic scalers affects the surfaces of composite restorative materials. Routine periodontal scaling should be carried out very carefully, and polishing of the scaled surfaces may overcome the alterations in roughness, thus preventing secondary caries, surface staining, plaque accumulation and subsequent periodontal inflammation. How to cite this article: Eid H A, Togoo R A, Saleh A A, Sumanth C R. Surface Topography of Composite Restorative Materials following Ultrasonic Scaling and its Impact on Bacterial Plaque Accumulation. An In-Vitro SEM Study. J Int Oral Health 2013; 5(3):13-19. PMID:24155597

A System for Photon-Counting Spectrophotometry of Prompt Optical Emission from Gamma-Ray Bursts

NASA Astrophysics Data System (ADS)

Vestrand, W. T.; Albright, K.; Casperson, D.; Fenimore, E.; Ho, C.; Priedhorsky, W.; White, R.; Wren, J.

2003-04-01

With the launch of HETE-2 and the coming launch of the Swift satellite, there will be many new opportunities to study the physics of the prompt optical emission with robotic ground-based telescopes. Time-resolved spectrophotometry of the rapidly varying optical emission is likely to be a rich area for discovery. We describe a program to apply state-of-the-art photon-counting imaging technology to the study of prompt optical emission from gamma-ray bursts. The Remote Ultra-Low Light Imaging (RULLI) project at Los Alamos National Laboratory has developed an imaging sensor which employs stacked microchannel plates and a crossed delay line readout with 200 picosecond photon timing to measure the time of arrival and positions for individual optical photons. RULLI detectors, when coupled with a transmission grating having 300 grooves/mm, can make photon-counting spectroscopic observations with spectral resolution that is an order of magnitude greater and temporal resolution three orders of magnitude greater than the most capable photon-counting imaging detectors that have been used for optical astronomy.

Label-free isolation and deposition of single bacterial cells from heterogeneous samples for clonal culturing

NASA Astrophysics Data System (ADS)

Riba, J.; Gleichmann, T.; Zimmermann, S.; Zengerle, R.; Koltay, P.

2016-09-01

The isolation and analysis of single prokaryotic cells down to 1 μm and less in size poses a special challenge and requires micro-engineered devices to handle volumes in the picoliter to nanoliter range. Here, an advanced Single-Cell Printer (SCP) was applied for automated and label-free isolation and deposition of bacterial cells encapsulated in 35 pl droplets by inkjet-like printing. To achieve this, dispenser chips to generate micro droplets have been fabricated with nozzles 20 μm in size. Further, the magnification of the optical system used for cell detection was increased. Redesign of the optical path allows for collision-free addressing of any flat substrate since no compartment protrudes below the nozzle of the dispenser chip anymore. The improved system allows for deterministic isolation of individual bacterial cells. A single-cell printing efficiency of 93% was obtained as shown by printing fluorescent labeled E. coli. A 96-well plate filled with growth medium is inoculated with single bacteria cells on average within about 8 min. Finally, individual bacterial cells from a heterogeneous sample of E. coli and E. faecalis were isolated for clonal culturing directly on agar plates in user-defined array geometry.

Bacterial pneumonia, HIV therapy, and disease progression among HIV-infected women in the HIV epidemiologic research (HER) study.

PubMed

Kohli, Rakhi; Lo, Yungtai; Homel, Peter; Flanigan, Timothy P; Gardner, Lytt I; Howard, Andrea A; Rompalo, Anne M; Moskaleva, Galina; Schuman, Paula; Schoenbaum, Ellie E

2006-07-01

To determine the rate and predictors of community-acquired bacterial pneumonia and its effect on human immunodeficiency virus (HIV) disease progression in HIV-infected women, we performed a multiple-site, prospective study of HIV-infected women in 4 cities in the United States. During the period of 1993-2000, we observed 885 HIV-infected and 425 HIV-uninfected women with a history of injection drug use or high-risk sexual behavior. Participants underwent semiannual interviews, and CD4+ lymphocyte count and viral load were assessed in HIV-infected subjects. Data regarding episodes of bacterial pneumonia were ascertained from medical record reviews. The rate of bacterial pneumonia among 885 HIV-infected women was 8.5 cases per 100 person-years, compared with 0.7 cases per 100 person-years in 425 HIV-uninfected women (P < .001). In analyses limited to follow-up after 1 January 1996, highly active antiretroviral therapy (HAART) and trimethoprim-sulfamethoxazole (TMP-SMX) use were associated with a decreased risk of bacterial pneumonia. Among women who had used TMP-SMX for 12 months, each month of HAART decreased bacterial pneumonia risk by 8% (adjusted hazard ratio [HR(adj)], 0.92; 95% confidence interval [CI], 0.89-0.95). Increments of 50 CD4+ cells/mm3 decreased the risk (HR(adj), 0.88; 95% CI, 0.84-0.93), and smoking doubled the risk (HR(adj), 2.12; 95% CI, 1.26-3.55). Bacterial pneumonia increased mortality risk (HR(adj), 5.02; 95% CI, 2.12-11.87), with adjustment for CD4+ lymphocyte count and duration of HAART and TMP-SMX use. High rates of bacterial pneumonia persist among HIV-infected women. Although HAART and TMP-SMX treatment decreased the risk, bacterial pneumonia was associated with an accelerated progression to death. Interventions that improve HAART utilization and promote smoking cessation among HIV-infected women are warranted.

Kids Count in Delaware, Families Count in Delaware: Fact Book, 2003.

ERIC Educational Resources Information Center

Delaware Univ., Newark. Kids Count in Delaware.

This Kids Count Fact Book is combined with the Families Count Fact Book to provide information on statewide trends affecting children and families in Delaware. The Kids Count and Families Count indicators have been combined into four new categories: health and health behaviors, educational involvement and achievement, family environment and…

DETECTION OF BACTERIAL CYTOTOXIC ACTIVITIES FROM WATER-DAMAGED CEILING TILE MATERIAL FOLLOWING INCUBATION ON BLOOD AGAR

EPA Science Inventory

Samples of ceiling tiles with high levels of bacteria exhibited cytotoxic activities on a HEP-2 tissue culture assay. Ceiling tiles containing low levels of bacterial colonization did not show cytotoxic activities on the HEP-2 tissue culture assay. Using a spread plate procedure ...

Duodenal bacterial overgrowth during treatment in outpatients with omeprazole.

PubMed Central

Fried, M; Siegrist, H; Frei, R; Froehlich, F; Duroux, P; Thorens, J; Blum, A; Bille, J; Gonvers, J J; Gyr, K

1994-01-01

The extent of duodenal bacterial overgrowth during the pronounced inhibition of acid secretion that occurs with omeprazole treatment is unknown. The bacterial content of duodenal juice of patients treated with omeprazole was therefore examined in a controlled prospective study. Duodenal juice was obtained under sterile conditions during diagnostic upper endoscopy. Aspirates were plated quantitatively for anaerobic and aerobic organisms. Twenty five outpatients with peptic ulcer disease were investigated after a 5.7 (0.5) weeks (mean (SEM)) treatment course with 20 mg (nine patients) or 40 mg (16 patients). The control group consisted of 15 outpatients referred for diagnostic endoscopy without prior antisecretory treatment. No patient in the control group had duodenal bacterial overgrowth. In the omeprazole group bacterial overgrowth (> or = 10(5) cfu/ml) was found in 14 (56%) patients (p = 0.0003). The number of bacteria (log10) in duodenal juice in patients treated with omeprazole was distinctly higher (median 5.7; range < 2-8.7) when compared with the control group (median < 2; range < 2-5.0; p = 0.0004). As well as orally derived bacteria, faecal type bacteria were found in seven of 14 and anaerobic bacteria in three of 14 patients. Bacterial overgrowth was similar with the two doses of omeprazole. These results indicate that duodenal bacterial overgrowth of both oral and faecal type bacteria occurs often in ambulatory patients treated with omeprazole. Further studies are needed to determine the clinical significance of these findings, particularly in high risk groups during long term treatment with omeprazole. PMID:8307444

Composite and Component Plates, Plate Non-rigidity, and the Steadiness of Plate Motion From Marine Geophysical and Space Geodetic Data

NASA Astrophysics Data System (ADS)

Gordon, R. G.; Argus, D. F.; DeMets, C.

2017-12-01

Plate tectonic theory has evolved since its birth 50 years ago. In particular, we now recognize that some of the originally proposed plates such as the Indo-Australia plate, the Africa plate, and the America plate are what we term "composite" plates—entities that contain no traditionally defined narrow plate boundaries, but are composed of multiple approximately rigid regions, which we term "component" plates, separated by diffuse plate boundaries. The best example of a composite plate is the Indo-Australia composite plate, which consists of the India, Capricorn, Australia, and Macquarie component plates and multiple intervening diffuse oceanic plate boundaries. The poles of relative rotation between component plates tend to lie in their mutual diffuse plate boundary. Outside of diffuse boundaries, plate rigidity has proven to be an excellent approximation, but the non-closure of some plate circuits indicates that stable plate interiors have a small but significant non-rigidity that may add up to 1 to 2 mm/a across any individual plate and may be partly due to horizontal thermal contraction of oceanic lithosphere. The greatest observational challenge to plate rigidity is posed by the Pacific-Cocos-Nazca plate circuit, which fails closure by 15 ±4 mm/a. The most rapid deformation of the plates observed with space geodesy is generated by solid Earth's viscous response to unloading of the late Pleistocene ice sheets. Differences between different realizations of global plate velocities from space geodesy appear in some cases to be due to differing assumptions about the motion of the geocenter, which affects estimated plate relative angular velocities and estimated vertical motion at geodetic sites. Comparison of space geodetic and marine geophysical plate motion rates and directions has demonstrated that plate motion is nearly steady, which allows plate boundary conditions to be applied to inter-seismic strain accumulation due to locking of specific faults. In

Absolute Enumeration of Probiotic Strains Lactobacillus acidophilus NCFM® and Bifidobacterium animalis subsp. lactis Bl-04 ® via Chip-Based Digital PCR.

PubMed

Hansen, Sarah J Z; Morovic, Wesley; DeMeules, Martha; Stahl, Buffy; Sindelar, Connie W

2018-01-01

The current standard for enumeration of probiotics to obtain colony forming units by plate counts has several drawbacks: long time to results, high variability and the inability to discern between bacterial strains. Accurate probiotic cell counts are important to confirm the delivery of a clinically documented dose for its associated health benefits. A method is described using chip-based digital PCR (cdPCR) to enumerate Bifidobacterium animalis subsp. lactis Bl-04 and Lactobacillus acidophilus NCFM both as single strains and in combination. Primers and probes were designed to differentiate the target strains against other strains of the same species using known single copy, genetic differences. The assay was optimized to include propidium monoazide pre-treatment to prevent amplification of DNA associated with dead probiotic cells as well as liberation of DNA from cells with intact membranes using bead beating. The resulting assay was able to successfully enumerate each strain whether alone or in multiplex. The cdPCR method had a 4 and 5% relative standard deviation (RSD) for Bl-04 and NCFM, respectively, making it more precise than plate counts with an industry accepted RSD of 15%. cdPCR has the potential to replace traditional plate counts because of its precision, strain specificity and the ability to obtain results in a matter of hours.

Comparison of the compact dry TC method with the standard method ISO 21149:2006 for determining aerobic colony counts in cosmetic emulsion.

PubMed

De Vaugelade, S; Aime, M; Farcette, N; Maurel, E; Lacour, T; Thomas, C; Bouchonnet, S; Pirnay, S

2017-02-01

Compact Dry TC, a rapid method kit for determining aerobic colony counts, has been developed by Nissui Pharmaceutical Co. for food application. These plates are pre-sterilized and contain culture medium, a cold-soluble gelling agent and a colour redox indicator for rapid enumeration. In this study, the alternative method is compared with the standard method ISO 21149:2006 - Cosmetic - Microbiology - Enumeration and detection of aerobic mesophilic bacteria, for cosmetic emulsions application. An oil-in-water (o/w) cosmetic emulsion was contaminated with a pool of bacterial strains (Escherichia coli ATCC 8739, Staphylococcus aureus ATCC 6538 and Pseudomonas aeruginosa ATCC 9027). One millilitre of samples was spread on agar as described in ISO 21149. The colonies were enumerated after 3 days of incubation. At the same time, 1.2 mL samples were spread on Compact Dry TC kits. The kit was incubated at 35°C ± 1°C for 48 h, and the colonies were enumerated. Accuracy determination was carried out using six replicates at four levels of concentrations (10, 50, 100 and 250 CFU mL -1 ). The repeatability study was carried out using 12 replicates at four levels of concentrations (10, 50, 100 and 250 CFU mL -1 ). Variations relative to the analyst and to the batch of emulsion have been investigated. The linear correlation coefficients of Compact Dry TC Kit enumeration with standard method ISO 21149:2006 was 0.9999. In comparison study, no apparent differences were noted between the Compact Dry TC kit and the reference method ISO 21149, for the detection level of aerobic microorganisms. Relative accuracy, repeatability and intermediate precision studies were acceptable. In the repeatability study, the Shapiro-Wilk test has confirmed the normally distribution of the twelve assays. No significant variations in Compact Dry TC count results were observed with different analysts and different batches of emulsion. The results showed that the two compared methods 'Compact Dry TC' vs

Rainfall simulation in greenhouse microcosms to assess bacterial-associated runoff from land-applied poultry litter.

PubMed

Brooks, John P; Adeli, Ardeshir; Read, John J; McLaughlin, Michael R

2009-01-01

Runoff water following a rain event is one possible source of environmental contamination after a manure application. This greenhouse study used a rainfall simulator to determine bacterial-associated runoff from troughs of common bermudagrass [Cynodon dactylon (L.) Pers.] that were treated with P-based, N-based, and N plus lime rates of poultry (Gallus gallus) litter, recommended inorganic fertilizer, and control. Total heterotrophic plate count (HPC) bacteria, total and thermotolerant coliforms, enterococci, staphylococci, Clostridium perfringens, Salmonella, and Campylobacter, as well as antibiotic resistance profiles for the staphylococci and enterococci isolates were all monitored in runoff waters. Analysis following five rainfall events indicated that staphylococci, enterococci, and clostridia levels were related to manure application rate. Runoff release of staphylococci, enterococci, and C. perfringens were approximately 3 to 6 log10 greater in litter vs. control treatment. In addition, traditional indicators such as thermotolerant and total coliforms performed poorly as fecal indicators. Some isolated enterococci demonstrated increased antibiotic resistance to polymixin b and/or select aminoglyocosides, while many staphylococci were susceptible to most antimicrobials tested. Results indicated poultry litter application can lead to microbial runoff following simulated rain events. Future studies should focus on the use of staphylococci, enterococci, and C. perfringens as indicators.

Incidence and Predictors of Bacterial infection in Febrile Children with Sickle Cell Disease.

PubMed

Morrissey, Benita J; Bycroft, Thomas P; Almossawi, Ofran; Wilkey, Olufunke B; Daniels, Justin G

2015-01-01

Children with sickle cell disease are at increased risk of developing bacteremia and other serious bacterial infections. Fever is a common symptom in sickle cell disease and can also occur with sickle cell crises and viral infections. We aimed to evaluate the incidence and predictors of bacteremia and bacterial infection in children with sickle cell disease presenting with fever to a district hospital and sickle cell center in London. A retrospective analysis was performed on all attendances of children (aged under 16 years) with sickle cell disease presenting with a fever of 38.5 °C or higher over a 1-year period. Confirmed bacterial infection was defined as bacteremia, bacterial meningitis, urinary tract infection (UTI), pneumonia, osteomyelitis or other bacterial infection with positive identification of organism. Children were defined as having a suspected bacterial infection if a bacterial infection was suspected clinically, but no organism was identified. Over a 1-year period there were 88 episodes analyzed in 59 children. Bacteremia occurred in 3.4% of episodes and confirmed bacterial infection in 7.0%. Suspected bacterial infection occurred in 33.0%. One death occurred from Salmonella typhirium septicemia. C-reactive protein (CRP) level and white blood cell (WBC) count were both significantly associated with bacterial infection (p = 0.004 and 0.02, respectively.) In conclusion, bacterial infections continue to be a significant problem in children with sickle cell disease. C-reactive protein was significantly associated with bacterial infections, and could be included in clinical risk criteria for febrile children with sickle cell disease.

Copper effects on bacterial activity of estuarine silty sediments

NASA Astrophysics Data System (ADS)

Almeida, Adelaide; Cunha, Ângela; Fernandes, Sandra; Sobral, Paula; Alcântara, Fernanda

2007-07-01

Bacteria of silty estuarine sediments were spiked with copper to 200 μg Cu g -1 dry weight sediment in order to assess the impact of copper on bacterial degradation of organic matter and on bacterial biomass production. Bacterial density was determined by direct counting under epifluorescence microscopy and bacterial production by the incorporation of 3H-Leucine. Leucine turnover rate was evaluated by 14C-leucine incorporation and ectoenzymatic activities were estimated as the hydrolysis rate of model substrates for β-glucosidase and leucine-aminopeptidase. The presence of added copper in the microcosms elicited, after 21 days of incubation, generalised anoxia and a decrease in organic matter content. The non-eroded surface of the copper-spiked sediment showed, when compared to the control, a decrease in bacterial abundance and significant lower levels of bacterial production and of leucine turnover rate. Bacterial production and leucine turnover rate decreased to 1.4% and 13% of the control values, respectively. Ectoenzymatic activities were also negatively affected but by smaller factors. After erosion by the water current in laboratory flume conditions, the eroded surface of the control sediment showed a generalised decline in all bacterial activities. The erosion of the copper-spiked sediment showed, however, two types of responses with respect to bacterial activities at the exposed surface: positive responses of bacterial production and leucine turnover rate contrasting with slight negative responses of ectoenzymatic activities. The effects of experimental erosion in the suspended cells were also different in the control and in the copper-spiked sediment. Bacterial cells in the control microcosm exhibited, when compared to the non-eroded sediment cells, decreases in all activities after the 6-h suspension. The response of the average suspended copper-spiked sediment cell differed from the control by a less sharp decrease in ectoenzymatic activities and

Short communication: A comparison of biofilm development on stainless steel and modified-surface plate heat exchangers during a 17-h milk pasteurization run.

PubMed

Jindal, Shivali; Anand, Sanjeev; Metzger, Lloyd; Amamcharla, Jayendra

2018-04-01

Flow of milk through the plate heat exchanger (PHE) results in denaturation of proteins, resulting in fouling. This also accelerates bacterial adhesion on the PHE surface, eventually leading to the development of biofilms. During prolonged processing, these biofilms result in shedding of bacteria and cross-contaminate the milk being processed, thereby limiting the duration of production runs. Altering the surface properties of PHE, such as surface energy and hydrophobicity, could be an effective approach to reduce biofouling. This study was conducted to compare the extent of biofouling on native stainless steel (SS) and modified-surface [Ni-P-polytetrafluoroethylene (PTFE)] PHE during the pasteurization of raw milk for an uninterrupted processing run of 17 h. For microbial studies, raw and pasteurized milk samples were aseptically collected from inlets and outlets of both PHE at various time intervals to examine shedding of bacteria in the milk. At the end of the run, 3M quick swabs (3M, St. Paul, MN) and ATP swabs (Charm Sciences Inc., Lawrence, MA) were used to sample plates from different sections of the pasteurizers (regeneration, heating, and cooling) for biofilm screening and to estimate the efficiency of cleaning in place, respectively. The data were tested for ANOVA, and means were compared. Modified PHE experienced lower mesophilic and thermophilic bacterial attachment and biofilm formation (average log 1.0 and 0.99 cfu/cm 2 , respectively) in the regenerative section of the pasteurizer compared with SS PHE (average log 1.49 and 1.47, respectively). Similarly, higher relative light units were observed for SS PHE compared with the modified PHE, illustrating the presence of more organic matter on the surface of SS PHE at the end of the run. In addition, at h 17, milk collected from the outlet of SS PHE showed plate counts of 5.44 cfu/cm 2 , which were significantly higher than those for pasteurized milk collected from modified PHE (4.12 log cfu/cm 2 ). This

A Microbial Community in Sediments Beneath the Western Antarctic Ice Sheet, Ice Stream C (Kamb)

NASA Astrophysics Data System (ADS)

Skidmore, M.; Han, S.; Foo, W.; Bui, D.; Lanoil, B.

2004-12-01

In 2000, an ice-drilling project focusing on the "sticky spot" of Ice Stream C recovered cores of sub-glacial sediments from beneath the Western Antarctic Ice Sheet. We have characterized several chemical and microbiological parameters of the sole intact sediment core. Pore waters extracted from these sediments were brackish and some were supersaturated with respect to calcite. Ion chromatography demonstrated the presence of several organic acids at low, but detectable, levels in the pore water. DAPI direct cell counts were approximately 107 cells g-1. Aerobic viable plate counts were much lower than direct cell counts; however, they were two orders of magnitude higher on plates incubated at low temperature (4 ° C; 3.63 x 105 CFU ml-1) than at higher temperatures (ca. 22° C; 1.5 x 103 CFU ml-1); no colonies were detected on plates incubated anaerobically at either temperature. 16S rDNA clone library analysis indicates extremely limited bacterial diversity in these samples: six phylogenetic clades were detected. The three dominant bacterial phylogenetic clades in the clone libraries (252 clones total) were most closely related to Thiobacillus thioparus (180 clones), Polaromonas vacuolata (34 clones), and Gallionella ferruginea (35 clones) and their relatives; one clone each represented the other three phylogenetic clades (most closely related to Ralstonia pickettii, Lysobacter antibioticus, and Xylella fastidiosa, respectively). These sequences match closely with sequences previously obtained from other subglacial environments in Alaska, Ellesmere Island, Canada and New Zealand. Implications of this microbial community to subglacial chemistry and microbial biogeography will be discussed.

Tower counts

USGS Publications Warehouse

Woody, Carol Ann; Johnson, D.H.; Shrier, Brianna M.; O'Neal, Jennifer S.; Knutzen, John A.; Augerot, Xanthippe; O'Neal, Thomas A.; Pearsons, Todd N.

2007-01-01

Counting towers provide an accurate, low-cost, low-maintenance, low-technology, and easily mobilized escapement estimation program compared to other methods (e.g., weirs, hydroacoustics, mark-recapture, and aerial surveys) (Thompson 1962; Siebel 1967; Cousens et al. 1982; Symons and Waldichuk 1984; Anderson 2000; Alaska Department of Fish and Game 2003). Counting tower data has been found to be consistent with that of digital video counts (Edwards 2005). Counting towers do not interfere with natural fish migration patterns, nor are fish handled or stressed; however, their use is generally limited to clear rivers that meet specific site selection criteria. The data provided by counting tower sampling allow fishery managers to determine reproductive population size, estimate total return (escapement + catch) and its uncertainty, evaluate population productivity and trends, set harvest rates, determine spawning escapement goals, and forecast future returns (Alaska Department of Fish and Game 1974-2000 and 1975-2004). The number of spawning fish is determined by subtracting subsistence, sport-caught fish, and prespawn mortality from the total estimated escapement. The methods outlined in this protocol for tower counts can be used to provide reasonable estimates ( plus or minus 6%-10%) of reproductive salmon population size and run timing in clear rivers. 

Radiation-hard ceramic Resistive Plate Chambers for forward TOF and T0 systems

NASA Astrophysics Data System (ADS)

Akindinov, A.; Dreyer, J.; Fan, X.; Kämpfer, B.; Kiselev, S.; Kotte, R.; Garcia, A. Laso; Malkevich, D.; Naumann, L.; Nedosekin, A.; Plotnikov, V.; Stach, D.; Sultanov, R.; Voloshin, K.

2017-02-01

Resistive Plate Chambers with ceramic electrodes are the main candidates for a use in precise multi-channel timing systems operating in high-radiation conditions. We report the latest R&D results on these detectors aimed to meet the requirements of the forward T0 counter at the CBM experiment. RPC design, gas mixture, limits on the bulk resistivity of ceramic electrodes, efficiency, time resolution, counting rate capabilities and ageing test results are presented.

Effect of omeprazole on intragastric bacterial counts, nitrates, nitrites, and N-nitroso compounds.

PubMed Central

Verdu, E; Viani, F; Armstrong, D; Fraser, R; Siegrist, H H; Pignatelli, B; Idström, J P; Cederberg, C; Blum, A L; Fried, M

1994-01-01

Previous studies have suggested that profound inhibition of gastric acid secretion may increase exposure to potentially carcinogenic N-nitroso compounds. The aim of this study was to find out if the proton pump inhibitor omeprazole (20 mg daily) is associated with increased concentrations of potentially carcinogenic N-nitroso compounds in gastric juice. The volume of gastric contents, number of bacteria, and concentrations of nitrates, nitrites, and N-nitroso compounds was determined in gastric aspirates obtained after an overnight fast in 14 healthy volunteers (7M:7F) after one week of treatment with placebo, and one and two weeks' treatment with omeprazole. Median bacterial concentrations were 1.0 x 10(4) (range 5.0 x 10(3)-5.0 x 10(6)) colony forming units (CFU)/ml after one weeks' treatment with placebo and increased significantly to 4.0 x 10(5) (0-3.3 x 10(7)) CFU/ml after two weeks' treatment with omeprazole (p < 0.05). A similar increase was seen in the concentration of nitrate reducing bacteria. There was no difference in the volume of gastric aspirates after treatment with omeprazole when compared with placebo (65 (29-155) ml v 42 (19-194) ml). The concentration of N-nitroso compounds was 0.13 (0-1.0) mumol/l after two weeks of omeprazole, which was not significantly different from that seen with placebo (0.15 (0-0.61) mumol/l). There was also no increase in the concentrations of nitrates or nitrites. It is concluded that omeprazole (20 mg once daily) for two weeks in healthy volunteers is associated with gastric bacterial proliferation but does not increase concentrations of N-nitroso compounds. PMID:8174980

Chronic bacterial prostatitis in men with spinal cord injury.

PubMed

Krebs, Jörg; Bartel, Peter; Pannek, Jürgen

2014-12-01

Recurrent urinary tract infections (UTI) are a major problem affecting spinal cord injury (SCI) patients and may stem from chronic bacterial prostatitis. We have therefore investigated the presence of chronic bacterial prostatitis and its role in the development of recurrent symptomatic UTI in SCI men. This study is a prospective cross-sectional investigation of bacterial prostatitis in SCI men in a single SCI rehabilitation center. In 50 men with chronic SCI presenting for a routine urologic examination, urine samples before and after prostate massage were taken for microbiologic investigation and white blood cell counting. Furthermore, patient characteristics, bladder diary details, and the annual rate of symptomatic UTI were collected retrospectively. No participant reported current symptoms of UTI or prostatitis. In most men (39/50, 78 %), the microbiologic analysis of the post-massage urine sample revealed growth of pathogenic bacteria. The majority of these men (32/39, 82 %) also presented with mostly (27/39, 69 %) the same pathogenic bacteria in the pre-massage sample. There was no significant (p = 0.48) difference in the number of symptomatic UTI in men with a positive post-massage culture compared with those with a negative culture. No significant (p = 0.67) difference in the frequency distribution of positive versus negative post-massage cultures was detected between men with recurrent and sporadic UTI. Most SCI men are affected by asymptomatic bacterial prostatitis; however, bacterial prostatitis does not play a major role in the development of recurrent UTI. The indication for antibiotic treatment of chronic bacterial prostatitis in asymptomatic SCI men with recurrent UTI is questionable.

Bacterial adhesion affinities of various implant abutment materials.

PubMed

Yamane, Koichi; Ayukawa, Yasunori; Takeshita, Toru; Furuhashi, Akihiro; Yamashita, Yoshihisa; Koyano, Kiyoshi

2013-12-01

To investigate bacterial adhesion to various abutment materials. Thirty volunteers participated in this study. Resin splints were fabricated, and five types of disks were fabricated from pure titanium, gold-platinum alloy, zirconia, alumina, and hydroxyapatite with uniform surface roughness and attached to the buccal surface of each splint. After 4 days of use by the subjects, the plaque accumulated on the disk surfaces was analyzed. The bacterial community structure was evaluated using 16S rRNA gene profiling with terminal restriction fragment length polymorphism analysis. The total bacterial count on each disk was estimated using quantitative polymerase chain reaction. Terminal restriction fragment length polymorphism profiles were more similar between tested materials than between subjects, suggesting that the bacterial community structures on the abutment material were influenced more by the individuals than by the type of material. However, the total number of bacteria attached to a disk was significantly different among five materials (P < 0.001, Brunner-Langer test for longitudinal data). Fewer bacteria were attached to the gold-platinum alloy than to the other materials. Gold-platinum alloy appears to be useful material for abutments when considering the accumulation of plaque. However, alternative properties of the abutment material, such as effects on soft tissue healing, should also be taken into consideration when choosing an abutment material. © 2012 John Wiley & Sons A/S.

MRP8/14 Enhances Corneal Susceptibility to Pseudomonas aeruginosa Infection by Amplifying Inflammatory Responses

PubMed Central

Deng, Qiuchan; Sun, Mingxia; Yang, Kun; Zhu, Min; Chen, Kang; Yuan, Jin; Wu, Minhao; Huang, Xi

2013-01-01

Purpose. We explored the role of myeloid-related protein 8 and 14 (MRP8/14) in Pseudomonas aeruginosa (PA) keratitis. Methods. MRP8/14 mRNA levels in human corneal scrapes and mouse corneas infected by PA were tested using real-time PCR. MRP8/14 protein expression in C57BL/6 (B6) corneas was confirmed using Western blot assay and immunohistochemistry. B6 mice were injected subconjunctivally with siRNA for MRP8/14, and then infected with PA. Bacterial plate counts and myeloperoxidase assays were used to determine the bacterial load and polymorphonuclear neutrophil (PMN) infiltration in infected B6 corneas. Pro-inflammatory cytokine levels in vivo and in vitro were examined with PCR and ELISA. In murine macrophage-like RAW264.7 cells, phagocytosis and bacterial killing were assessed using plate count assays, and reactive oxygen species (ROS) and nitric oxide (NO) levels were tested with flow cytometry and Griess assay, respectively. Results. MRP8/14 expression levels were increased significantly in human corneal scrapes and B6 corneas after PA infection. Silencing of MRP8/14 in B6 corneas significantly reduced the severity of corneal disease, bacterial clearance, PMN infiltration, and pro-inflammatory cytokine expression after PA infection. In vitro studies demonstrated further that silencing of MRP8/14 suppressed pro-inflammatory cytokine production, bacterial killing, and ROS production, but not phagocytosis or NO production. Conclusions. Our study demonstrated a dual role for MRP8/14 in bacterial keratitis. Although MRP8/14 promotes bacterial clearance by enhancing ROS production, it functions more importantly as an inflammatory amplifier at the ocular surface by enhancing pro-inflammatory cytokine expression, thus contributing to the corneal susceptibility. PMID:23299480

Bacterial adhesion on direct and indirect dental restorative composite resins: An in vitro study on a natural biofilm.

PubMed

Derchi, Giacomo; Vano, Michele; Barone, Antonio; Covani, Ugo; Diaspro, Alberto; Salerno, Marco

2017-05-01

Both direct and indirect techniques are used for dental restorations. Which technique should be preferred or whether they are equivalent with respect to bacterial adhesion is unclear. The purpose of this in vitro study was to determine the affinity of bacterial biofilm to dental restorative composite resins placed directly and indirectly. Five direct composite resins for restorations (Venus Diamond, Adonis, Optifil, Enamel Plus HRi, Clearfil Majesty Esthetic) and 3 indirect composite resins (Gradia, Estenia, Signum) were selected. The materials were incubated in unstimulated whole saliva for 1 day. The biofilms grown were collected and their bacterial cells counted. In parallel, the composite resin surface morphology was analyzed with atomic force microscopy. Both bacterial cell count and surface topography parameters were subjected to statistical analysis (α=.05). Indirect composite resins showed significantly lower levels than direct composite resins for bacterial cell adhesion, (P<.001). No significant differences were observed within the direct composite resins (P>.05). However, within the indirect composite resins a significantly lower level was found for Gradia than Estenia or Signum (P<.01). A partial correlation was observed between composite resin roughness and bacterial adhesion when the second and particularly the third-order statistical moments of the composite resin height distributions were considered. Indirect dental restorative composite resins were found to be less prone to biofilm adhesion than direct composite resins. A correlation of bacterial adhesion to surface morphology exists that is described by kurtosis; thus, advanced data analysis is required to discover possible insights into the biologic effects of morphology. Copyright © 2016 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

Powerful colloidal silver nanoparticles for the prevention of gastrointestinal bacterial infections

NASA Astrophysics Data System (ADS)

Le, Anh-Tuan; Tam Le, Thi; Quy Nguyen, Van; Hoang Tran, Huy; Dang, Duc Anh; Tran, Quang Huy; Vu, Dinh Lam

2012-12-01

In this work we have demonstrated a powerful disinfectant ability of colloidal silver nanoparticles (NPs) for the prevention of gastrointestinal bacterial infections. The silver NPs colloid was synthesized by a UV-enhanced chemical precipitation. Two gastrointestinal bacterial strains of Escherichia coli (ATCC 43888-O157:k-:H7) and Vibrio cholerae (O1) were used to verify the antibacterial activity of the as-prepared silver NPs colloid by means of surface disinfection assay in agar plates and turbidity assay in liquid media. Transmission electron microscopy was also employed to analyze the ultrastructural changes of bacterial cells caused by silver NPs. Noticeably, our silver NPs colloid displayed a highly effective bactericidal effect against two tested gastrointestinal bacterial strains at a silver concentration as low as ˜3 mg l-1. More importantly, the silver NPs colloid showed an enhancement of antibacterial activity and long-lasting disinfectant effect as compared to conventional chloramin B (5%) disinfection agent. These advantages of the as-prepared colloidal silver NPs make them very promising for environmental treatments contaminated with gastrointestinal bacteria and other infectious pathogens. Moreover, the powerful disinfectant activity of silver-containing materials can also help in controlling and preventing further outbreak of diseases.

Normalization of test and evaluation of biothreat detection systems: overcoming microbial air content fluctuations by using a standardized reagent bacterial mixture.

PubMed

Berchebru, Laurent; Rameil, Pascal; Gaudin, Jean-Christophe; Gausson, Sabrina; Larigauderie, Guilhem; Pujol, Céline; Morel, Yannick; Ramisse, Vincent

2014-10-01

Test and evaluation of engineered biothreat agent detection systems ("biodetectors") are a challenging task for government agencies and industries involved in biosecurity and biodefense programs. In addition to user friendly features, biodetectors need to perform both highly sensitive and specific detection, and must not produce excessive false alerts. In fact, the atmosphere displays a number of variables such as airborne bacterial content that can interfere with the detection process, thus impeding comparative tests when carried out at different times or places. To overcome these bacterial air content fluctuations, a standardized reagent bacterial mixture (SRBM), consisting in a collection of selected cultivable environmental species that are prevalent in temperate climate bioaerosols, was designed to generate a stable, reproducible, and easy to use surrogate of bioaerosol sample. The rationale, design, and production process are reported. The results showed that 8.59; CI 95%: 8.46-8.72 log cfu distributed into vials underwent a 0.95; CI 95%: 0.65-1.26 log viability decay after dehydration and subsequent reconstitution, thus advantageously mimicking a natural bioaerosol sample which is typically composed of cultivable and uncultivable particles. Dehydrated SRBM was stable for more than 12months at 4°C and allowed the reconstitution of a dead/live cells aqueous suspension that is stable for 96h at +4°C, according to plate counts. Specific detection of a simulating biothreat agent (e.g. Bacillus atrophaeus) by immuno-magnetic or PCR assays did not display any significant loss of sensitivity, false negative or positive results in the presence of SRBM. This work provides guidance on testing and evaluating detection devices, and may contribute to the establishment of suitable standards and normalized procedures. Copyright © 2014 Elsevier B.V. All rights reserved.

Coupling MALDI-TOF mass spectrometry protein and specialized metabolite analyses to rapidly discriminate bacterial function

PubMed Central

Clark, Chase M.; Costa, Maria S.

2018-01-01

For decades, researchers have lacked the ability to rapidly correlate microbial identity with bacterial metabolism. Since specialized metabolites are critical to bacterial function and survival in the environment, we designed a data acquisition and bioinformatics technique (IDBac) that utilizes in situ matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to analyze protein and specialized metabolite spectra recorded from single bacterial colonies picked from agar plates. We demonstrated the power of our approach by discriminating between two Bacillus subtilis strains in <30 min solely on the basis of their differential ability to produce cyclic peptide antibiotics surfactin and plipastatin, caused by a single frameshift mutation. Next, we used IDBac to detect subtle intraspecies differences in the production of metal scavenging acyl-desferrioxamines in a group of eight freshwater Micromonospora isolates that share >99% sequence similarity in the 16S rRNA gene. Finally, we used IDBac to simultaneously extract protein and specialized metabolite MS profiles from unidentified Lake Michigan sponge-associated bacteria isolated from an agar plate. In just 3 h, we created hierarchical protein MS groupings of 11 environmental isolates (10 MS replicates each, for a total of 110 spectra) that accurately mirrored phylogenetic groupings. We further distinguished isolates within these groupings, which share nearly identical 16S rRNA gene sequence identity, based on interspecies and intraspecies differences in specialized metabolite production. IDBac is an attempt to couple in situ MS analyses of protein content and specialized metabolite production to allow for facile discrimination of closely related bacterial colonies. PMID:29686101

Effect of early antibiotic administration on cecal bacterial communities and their metabolic profiles in pigs fed diets with different protein levels.

PubMed

Zhang, Chuanjian; Yu, Miao; Yang, Yuxiang; Mu, Chunlong; Su, Yong; Zhu, Weiyun

2016-12-01

This study investigated the effects of early antibiotic administration (EAA) on cecal bacterial communities and their metabolic profiles in pigs fed diets with different protein levels. Eighteen litters (total 180) of piglets on day (d) 7 were fed either a commercial creep feed or commercial creep feed + antibiotic (Olaquindox, Oxytetracycline Calcium and Kitasamycin) until d 42. On d 42, pigs within each group were further randomly fed a normal crude protein (CP) diet (20% and 18% CP from d 42 to d 77 and d 77 to d 120, respectively) or a low-CP diet (16% and 14% CP from d 42 to d 77 and d 77 to d 120, respectively), generating 4 groups, control-low CP (Con-LP), control-normal CP (Con-NP), antibiotic-low CP (Ant-LP) and antibiotic-normal CP (Ant-NP), respectively. On d 77 and d 120, 5 pigs per group were slaughtered and cecal materials were collected for bacterial analysis. With cecal bacteria, principle component analysis (PCA) of the denaturing gradient gel electrophoresis (DGGE) profile showed two distinct groups of samples from low-CP diet and samples from normal-CP diet. Real-time PCR showed that EAA did not have significant effect on major bacterial groups, only showed significant interactions (P < 0.05) with CP level for Lactobacillus counts on d 77 and Clostridium cluster XIVa counts on d 120 with higher values in the Con-NP group compared to the Ant-NP groups. Low-CP diet increased (P < 0.05) short-chain fatty acids (SCFA) producing bacteria counts (Bacteroidetes on d 77 and d 120; Clostridium cluster IV and Clostridium cluster XIVa on d 77), but decreased (P < 0.05) Escherichia coli counts on d 77 and d 120. For metabolites, EAA increased (P < 0.05) protein fermentation products (p-cresol, indole and skatole on d 77; ammonia, putrescine and spermidine on d 120), and showed significant interactions (P < 0.05) with CP level for p-cresol and skatole concentrations on d 77 and putrescine and spermidine concentrations on d 120 with higher values

Changes in the bacterial community of soybean rhizospheres during growth in the field.

PubMed

Sugiyama, Akifumi; Ueda, Yoshikatsu; Zushi, Takahiro; Takase, Hisabumi; Yazaki, Kazufumi

2014-01-01

Highly diverse communities of bacteria inhabiting soybean rhizospheres play pivotal roles in plant growth and crop production; however, little is known about the changes that occur in these communities during growth. We used both culture-dependent physiological profiling and culture independent DNA-based approaches to characterize the bacterial communities of the soybean rhizosphere during growth in the field. The physiological properties of the bacterial communities were analyzed by a community-level substrate utilization assay with BioLog Eco plates, and the composition of the communities was assessed by gene pyrosequencing. Higher metabolic capabilities were found in rhizosphere soil than in bulk soil during all stages of the BioLog assay. Pyrosequencing analysis revealed that differences between the bacterial communities of rhizosphere and bulk soils at the phylum level; i.e., Proteobacteria were increased, while Acidobacteria and Firmicutes were decreased in rhizosphere soil during growth. Analysis of operational taxonomic units showed that the bacterial communities of the rhizosphere changed significantly during growth, with a higher abundance of potential plant growth promoting rhizobacteria, including Bacillus, Bradyrhizobium, and Rhizobium, in a stage-specific manner. These findings demonstrated that rhizosphere bacterial communities were changed during soybean growth in the field.

Bacteriological and genetic assessment of game meat from Japanese wild boars.

PubMed

Naya, Yuka; Horiuchi, Motohiro; Ishiguro, Naotaka; Shinagawa, Morikazu

2003-01-15

Bacterial tests were used to assess bacterial contamination of game meat from Japanese wild boars. The bacterial contamination of wild boar meat was less than that of domestic pork, as determined by aerobic plate counts (APC) and coliform counts. None of the meat examined in this study was contaminated by Salmonella or E. coli O-157. To detect adulteration by domestic pig meat or European wild boar meat, 46 samples of game meat sold as Japanese wild boar were examined genetically. A total of 17 samples showed genetic haplotypes of European and Asian domestic pigs in the D-loop of mitochondrial DNA (mtDNA), and 16 samples showed nuclear glucosephosphate isomerase-processed pseudogene (GPIP) genotypes of European domestic pigs. The European GPIP genotypes of these samples were confirmed by PCR-RFLP analysis. These results indicate that some game meat sold as Japanese wild boar is adulterated by cross-breeding between pigs and wild boars or by contamination with meat from domestic pigs or European wild boars.

Multiplicity Counting

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

Geist, William H.

2015-12-01

This set of slides begins by giving background and a review of neutron counting; three attributes of a verification item are discussed: 240Pu eff mass; α, the ratio of (α,n) neutrons to spontaneous fission neutrons; and leakage multiplication. It then takes up neutron detector systems – theory & concepts (coincidence counting, moderation, die-away time); detector systems – some important details (deadtime, corrections); introduction to multiplicity counting; multiplicity electronics and example distributions; singles, doubles, and triples from measured multiplicity distributions; and the point model: multiplicity mathematics.

Performance optimization of plate heat exchangers with chevron plates

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

Muley, A.; Manglik, R.M.

1999-07-01

The enhanced heat transfer performance of a chevron plate heat exchanger (PHE) is evaluated employing (1) energy-conservation based performance evaluation criteria (PECs), and (2) the second-law based minimization of entropy generation principle. Single-phase laminar and turbulent flow convection for three different chevron-plate arrangements are considered. The influence of plate surface corrugation characteristics and their stack arrangements on the heat exchanger's thermal-hydraulic performance is delineated. Based on the different figures of merit, the results show that the extent of heat transfer enhancement increases with flow Re and chevron angle {beta} in laminar flow, but it diminishes with increasing Re in turbulentmore » flows. With up to 2.9 times higher Q, 48% lower A, and entropy generation number N{sub s,a} {lt} 1, relative to an equivalent flat-plate pack, chevron plates are found to be especially suitable in the low to medium flow rates range (20 {le} Re {le} 2,000). Also, there appears to be no significant advantage of using a mixed-plate over a symmetric-plate arrangement.« less

Role of Clinical Presentations and Routine CSF Analysis in the Rapid Diagnosis of Acute Bacterial Meningitis in Cases of Negative Gram Stained Smears

PubMed Central

Fouad, Rabab; El-Kholy, Badawy; Yosry, Ayman

2014-01-01

Background and Aim. Bacterial meningitis is a lethal, disabling endemic disease needing prompt antibiotic management. Gram stained smears is rapid accurate method for diagnosis of bacterial meningitis. In cases of negative gram stained smears diagnosis is delayed till culture results. We aim to assess the role of clinical presentations and routine CSF analysis in the cost-effective rapid diagnosis of negative gram stained smears bacterial meningitis. Methods. Cross sectional study including 623 acute meningitis patients divided into two groups: bacterial meningitis and nonbacterial meningitis groups. The clinical presentations, systemic inflammatory parameters, and CSF analysis were evaluated and compared in both groups. Results. Altered conscious level, localizing neurological signs, Kernig's and Brudzinski's signs together with peripheral leucocytosis (>10.000/mm3), high CRP (>6) together with high CSF protein (>50 gl/dL), CSF neutrophilic count (≥50% of total CSF leucocytic count), and low CSF glucose level (<45 gm/dL) and CSF/serum glucose ≤0.6 were significantly diagnostic in bacterial meningitis patients. From the significant CSF analysis variables CSF protein carried the higher accuracy of diagnosis 78% with sensitivity 88% and specificity 72%. Conclusions. High CSF protein (>50 mg/dL) together with plasma inflammatory markers and CSF cytochemical parameters can diagnose bacterial meningitis in gram stain negative smear till culture results. PMID:24803939

Predictors of bacterial pneumonia in Evaluation of Subcutaneous Interleukin-2 in a Randomized International Trial (ESPRIT).

PubMed

Pett, S L; Carey, C; Lin, E; Wentworth, D; Lazovski, J; Miró, J M; Gordin, F; Angus, B; Rodriguez-Barradas, M; Rubio, R; Tambussi, G; Cooper, D A; Emery, S

2011-04-01

Bacterial pneumonia still contributes to morbidity/mortality in HIV infection despite effective combination antiretroviral therapy (cART). Evaluation of Subcutaneous Interleukin-2 in a Randomized International Trial (ESPRIT), a trial of intermittent recombinant interleukin-2 (rIL-2) with cART vs. cART alone (control arm) in HIV-infected adults with CD4 counts ≥300cells/μL, offered the opportunity to explore associations between bacterial pneumonia and rIL-2, a cytokine that increases the risk of some bacterial infections. Baseline and time-updated factors associated with first-episode pneumonia on study were analysed using multivariate proportional hazards regression models. Information on smoking/pneumococcal vaccination history was not collected. IL-2 cycling was most intense in years 1-2. Over ≈7 years, 93 IL-2 [rate 0.67/100 person-years (PY)] and 86 control (rate 0.63/100 PY) patients experienced a pneumonia event [hazard ratio (HR) 1.06; 95% confidence interval (CI) 0.79, 1.42; P=0.68]. Median CD4 counts prior to pneumonia were 570cells/μL (IL-2 arm) and 463cells/μL (control arm). Baseline risks for bacterial pneumonia included older age, injecting drug use, detectable HIV viral load (VL) and previous recurrent pneumonia; Asian ethnicity was associated with decreased risk. Higher proximal VL (HR for 1 log(10) higher VL 1.28; 95% CI 1.11, 1.47; P<0.001) was associated with increased risk; higher CD4 count prior to the event (HR per 100 cells/μL higher 0.94; 95% CI 0.89, 1.0; P=0.04) decreased risk. Compared with controls, the hazard for a pneumonia event was higher if rIL-2 was received <180 days previously (HR 1.66; 95% CI 1.07, 2.60; P=0.02) vs.≥180 days previously (HR 0.98; 95% CI 0.70, 1.37; P=0.9). Compared with the control group, pneumonia risk in the IL-2 arm decreased over time, with HRs of 1.41, 1.71, 1.16, 0.62 and 0.84 in years 1, 2, 3-4, 5-6 and 7, respectively. Bacterial pneumonia rates in cART-treated adults with moderate

Evaluation of finger plate and flat plate connection design.

DOT National Transportation Integrated Search

2016-01-01

This project investigates the cause(s) of premature deterioration of MoDOT finger plate and flat plate expansion devices : under high traffic volumes and then uses that information to design new Load and Resistance Factor Design (LRFD) : finger plate...

Details: Elevation of Plate Typical Bay, SectionThrough Plate Typical Bay, ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

Details: Elevation of Plate Typical Bay, Section-Through Plate Typical Bay, Section-Through Plate Center Bay, Elevation of Plate Center Bay - Contoocook Covered Bridge, Spanning Contoocook River, Hopkinton, Merrimack County, NH

Free Surface Effects on the Wake of a Flat Plate.

DTIC Science & Technology

1984-11-08

D-i46 98 FREE SURFCE’EFFECTS ON THE MAKE OF A FLAT PLTE(U) i/l 9(8 NAVAL RESEARCH LAB WASHINGTON DC T F SWEAN ET AL. 08 NOV 84 NRL-MR...5426UNCLASSIFIED F/ 20/4 NL 11111 ~ L.0 2 4 11111L .563 I -A 16 CEO -- . . IV NRL Memorandum Rpot52 Free Surface iEffwcs on the Wake of Al lit Plate T . F. SWEAlJ...13b. TIME COVERED 14. DATE OF REPORT (YeasrUonitDay) S.PAGE COUNT .0 - Interim IFROM _ TO T 1984 November 8 FS23 16 SUPPLEMENTARY NOTATION 17 COSATI

Changes in bacterial and archaeal communities during the concentration of brine at the graduation towers in Ciechocinek spa (Poland).

PubMed

Kalwasińska, Agnieszka; Deja-Sikora, Edyta; Burkowska-But, Aleksandra; Szabó, Attila; Felföldi, Támas; Kosobucki, Przemysław; Krawiec, Arkadiusz; Walczak, Maciej

2018-03-01

This study evaluates the changes in bacterial and archaeal community structure during the gradual evaporation of water from the brine (extracted from subsurface Jurassic deposits) in the system of graduation towers located in Ciechocinek spa, Poland. The communities were assessed with 16S rRNA gene sequencing (MiSeq, Illumina) and microscopic methods. The microbial cell density determined by direct cell count was at the order of magnitude of 10 7 cells/mL. It was found that increasing salt concentration was positively correlated with both the cell counts, and species-level diversity of bacterial and archaeal communities. The archaeal community was mostly constituted by members of the phylum Euryarchaeota, class Halobacteria and was dominated by Halorubrum-related sequences. The bacterial community was more diverse, with representatives of the phyla Proteobacteria and Bacteroidetes as the most abundant. The proportion of Proteobacteria decreased with increasing salt concentration, while the proportion of Bacteroidetes increased significantly in the more concentrated samples. Representatives of the genera Idiomarina, Psychroflexus, Roseovarius, and Marinobacter appeared to be tolerant to changes of salinity. During the brine concentration, the relative abundances of Sphingobium and Sphingomonas were significantly decreased and the raised contributions of genera Fabibacter and Fodinibius were observed. The high proportion of novel (not identified at 97% similarity level) bacterial reads (up to 42%) in the 16S rRNA gene sequences indicated that potentially new bacterial taxa inhabit this unique environment.

In vitro anti-biofilm and anti-bacterial activity of Junceella juncea for its biomedical application

PubMed Central

Kumar, P; Selvi, S Senthamil; Govindaraju, M

2012-01-01

Objective To investigate the anti-biofilm and anti-bacterial activity of Junceella juncea (J. juncea) against biofilm forming pathogenic strains. Methods Gorgonians were extracted with methanol and analysed with fourier transform infrared spectroscopy. Biofilm forming pathogens were identified by Congo red agar supplemented with sucrose. A quantitative spectrophotometric method was used to monitor in vitro biofilm reduction by microtitre plate assay. Anti-bacterial activity of methanolic gorgonian extract (MGE) was carried out by disc diffusion method followed by calculating the percentage of increase with crude methanol (CM). Results The presence of active functional group was exemplified by FT-IR spectroscopy. Dry, black, crystalline colonies confirm the production of extracellular polymeric substances responsible for biofilm formation in Congo red agar. MGE exhibited potential anti-biofilm activity against all tested bacterial strains. The anti-bacterial activity of methanolic extract was comparably higher in Salmonella typhii followed by Escherichia coli, Vibrio cholerae and Shigella flexneri. The overall percentage of increase was higher by 50.2% to CM. Conclusions To conclude, anti-biofilm and anti-bacterial efficacy of J. juncea is impressive over biofilm producing pathogens and are good source for novel anti-bacterial compounds. PMID:23593571

A Novel 96well-formatted Micro-gap Plate Enabling Drug Response Profiling on Primary Tumour Samples

NASA Astrophysics Data System (ADS)

Ma, Wei-Yuan; Hsiung, Lo-Chang; Wang, Chen-Ho; Chiang, Chi-Ling; Lin, Ching-Hung; Huang, Chiun-Sheng; Wo, Andrew M.

2015-04-01

Drug-based treatments are the most widely used interventions for cancer management. Personalized drug response profiling remains inherently challenging with low cell count harvested from tumour sample. We present a 96well-formatted microfluidic plate with built-in micro-gap that preserves up to 99.2% of cells during multiple assay/wash operation and only 9,000 cells needed for a single reagent test (i.e. 1,000 cells per test spot x 3 selected concentration x triplication), enabling drug screening and compatibility with conventional automated workstations. Results with MCF7 and MDA-MB-231 cell lines showed that no statistical significance was found in dose-response between the device and conventional 96-well plate control. Primary tumour samples from breast cancer patients tested in the device also showed good IC50 prediction. With drug screening of primary cancer cells must consider a wide range of scenarios, e.g. suspended/attached cell types and rare/abundant cell availability, the device enables high throughput screening even for suspended cells with low cell count since the signature microfluidic cell-trapping feature ensures cell preservation in a multiple solution exchange protocol.

Office space bacterial abundance and diversity in three metropolitan areas.

PubMed

Hewitt, Krissi M; Gerba, Charles P; Maxwell, Sheri L; Kelley, Scott T

2012-01-01

People in developed countries spend approximately 90% of their lives indoors, yet we know little about the source and diversity of microbes in built environments. In this study, we combined culture-based cell counting and multiplexed pyrosequencing of environmental ribosomal RNA (rRNA) gene sequences to investigate office space bacterial diversity in three metropolitan areas. Five surfaces common to all offices were sampled using sterile double-tipped swabs, one tip for culturing and one for DNA extraction, in 30 different offices per city (90 offices, 450 total samples). 16S rRNA gene sequences were PCR amplified using bar-coded "universal" bacterial primers from 54 of the surfaces (18 per city) and pooled for pyrosequencing. A three-factorial Analysis of Variance (ANOVA) found significant differences in viable bacterial abundance between offices inhabited by men or women, among the various surface types, and among cities. Multiplex pyrosequencing identified more than 500 bacterial genera from 20 different bacterial divisions. The most abundant of these genera tended to be common inhabitants of human skin, nasal, oral or intestinal cavities. Other commonly occurring genera appeared to have environmental origins (e.g., soils). There were no significant differences in the bacterial diversity between offices inhabited by men or women or among surfaces, but the bacterial community diversity of the Tucson samples was clearly distinguishable from that of New York and San Francisco, which were indistinguishable. Overall, our comprehensive molecular analysis of office building microbial diversity shows the potential of these methods for studying patterns and origins of indoor bacterial contamination. "[H]umans move through a sea of microbial life that is seldom perceived except in the context of potential disease and decay." - Feazel et al. (2009).

Development of a nematode offspring counting assay for rapid and simple soil toxicity assessment.

PubMed

Kim, Shin Woong; Moon, Jongmin; Jeong, Seung-Woo; An, Youn-Joo

2018-05-01

Since the introduction of standardized nematode toxicity assays by the American Society for Testing and Materials (ASTM) and International Organization for Standardization (ISO), many studies have reported their use. Given that the currently used standardized nematode toxicity assays have certain limitations, in this study, we examined the use of a novel nematode offspring counting assay for evaluating soil ecotoxicity based on a previous soil-agar isolation method used to recover live adult nematodes. In this new assay, adult Caenorhabditis elegans were exposed to soil using a standardized toxicity assay procedure, and the resulting offspring in test soils attracted by a microbial food source in agar plates were counted. This method differs from previously used assays in terms of its endpoint, namely, the number of nematode offspring. The applicability of the bioassay was demonstrated using metal-spiked soils, which revealed metal concentration-dependent responses, and with 36 field soil samples characterized by different physicochemical properties and containing various metals. Principal component analysis revealed that texture fraction (clay, sand, and silt) and electrical conductivity values were the main factors influencing the nematode offspring counting assay, and these findings warrant further investigation. The nematode offspring counting assay is a rapid and simple process that can provide multi-directional toxicity assessment when used in conjunction with other standard methods. Copyright © 2018 Elsevier Ltd. All rights reserved.

Gradient microfluidics enables rapid bacterial growth inhibition testing.

PubMed

Li, Bing; Qiu, Yong; Glidle, Andrew; McIlvenna, David; Luo, Qian; Cooper, Jon; Shi, Han-Chang; Yin, Huabing

2014-03-18

Bacterial growth inhibition tests have become a standard measure of the adverse effects of inhibitors for a wide range of applications, such as toxicity testing in the medical and environmental sciences. However, conventional well-plate formats for these tests are laborious and provide limited information (often being restricted to an end-point assay). In this study, we have developed a microfluidic system that enables fast quantification of the effect of an inhibitor on bacteria growth and survival, within a single experiment. This format offers a unique combination of advantages, including long-term continuous flow culture, generation of concentration gradients, and single cell morphology tracking. Using Escherichia coli and the inhibitor amoxicillin as one model system, we show excellent agreement between an on-chip single cell-based assay and conventional methods to obtain quantitative measures of antibiotic inhibition (for example, minimum inhibition concentration). Furthermore, we show that our methods can provide additional information, over and above that of the standard well-plate assay, including kinetic information on growth inhibition and measurements of bacterial morphological dynamics over a wide range of inhibitor concentrations. Finally, using a second model system, we show that this chip-based systems does not require the bacteria to be labeled and is well suited for the study of naturally occurring species. We illustrate this using Nitrosomonas europaea, an environmentally important bacteria, and show that the chip system can lead to a significant reduction in the period required for growth and inhibition measurements (<4 days, compared to weeks in a culture flask).

Bacterial and primary production in the pelagic zone of the Kara Sea

NASA Astrophysics Data System (ADS)

Sazhin, A. F.; Romanova, N. D.; Mosharov, S. A.

2010-10-01

Data on the bacterial and primary production, which were obtained simultaneously for the same water samples, are presented for three regions of the Kara Sea. The samples were collected for the transect westwards of the Yamal Peninsula, along the St. Anna Trough, and the transect in Ob Bay. Direct counts of the DAPI-stained bacterial cells were performed. The bacterial production and grazing rates were determined using a direct method when metabolic inhibitors vancomycin and penicillin were added. The primary production rates were estimated using the 14C method. The average primary production was 112.6, 58.5, and 28.7 mg C m-2 day-1, and the bacterial production was 12.8, 48.9, and 81.6 mg C m-2 day-1 along the Yamal Peninsula, the St. Anna Trough, and Ob Bay, respectively. The average bacterial carbon demand was 34.6, 134.5, and 220.4 mg C m-2 day-1 for these regions, respectively. The data obtained lead us to conclude that the phytoplankton-synthesized organic matter is generally insufficient to satisfy the bacterial carbon demand and may be completely assimilated via the heterotrophic processes in the marine ecosystems. Therefore, the bacterial activity and, consequently, the amount of the synthesized biomass (i.e., the production) both depend directly on the phytoplankton’s condition and activity. We consider these relationships to be characteristics of the Kara Sea’s biota.

Diagnostic value of lactate, procalcitonin, ferritin, serum-C-reactive protein, and other biomarkers in bacterial and viral meningitis: A cross-sectional study.

PubMed

Sanaei Dashti, Anahita; Alizadeh, Shekoofan; Karimi, Abdullah; Khalifeh, Masoomeh; Shoja, Seyed Abdolmajid

2017-09-01

There are many difficulties distinguishing bacterial from viral meningitis that could be reasonably solved using biomarkers. The aim of this study was to evaluate lactate, procalcitonin (PCT), ferritin, serum-CRP (C-reactive protein), and other known biomarkers in differentiating bacterial meningitis from viral meningitis in children.All children aged 28 days to 14 years with suspected meningitis who were admitted to Mofid Children's Hospital, Tehran, between October 2012 and November 2013, were enrolled in this prospective cross-sectional study. Children were divided into 2 groups of bacterial and viral meningitis, based on the results of cerebrospinal fluid (CSF) culture, polymerase chain reaction, and cytochemical profile. Diagnostic values of CSF parameters (ferritin, PCT, absolute neutrophil count [ANC], white blood cell count, and lactate) and serum parameters (PCT, ferritin, CRP, and erythrocyte sedimentation rate [ESR]) were evaluated.Among 50 patients with meningitis, 12 were diagnosed with bacterial meningitis. Concentrations of all markers were significantly different between bacterial and viral meningitis, except for serum (P = .389) and CSF (P = .136) PCT. The best rates of area under the receiver operating characteristic (ROC) curve (AUC) were achieved by lactate (AUC = 0.923) and serum-CRP (AUC = 0.889). The best negative predictive values (NPV) for bacterial meningitis were attained by ANC (100%) and lactate (97.1%).The results of our study suggest that ferritin and PCT are not strong predictive biomarkers. A combination of low CSF lactate, ANC, ESR, and serum-CRP could reasonably rule out the bacterial meningitis.