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

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Stefanie Hoffmann

2018-02-01

Full Text Available 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.

Short communication: Repeatability of differential goat bulk milk culture and associations with somatic cell count, total bacterial count, and standard plate count.

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Koop, G; Dik, N; Nielen, M; Lipman, L J A

2010-06-01

The aims of this study were to assess how different bacterial groups in bulk milk are related to bulk milk somatic cell count (SCC), bulk milk total bacterial count (TBC), and bulk milk standard plate count (SPC) and to measure the repeatability of bulk milk culturing. On 53 Dutch dairy goat farms, 3 bulk milk samples were collected at intervals of 2 wk. The samples were cultured for SPC, coliform count, and staphylococcal count and for the presence of Staphylococcus aureus. Furthermore, SCC (Fossomatic 5000, Foss, Hillerød, Denmark) and TBC (BactoScan FC 150, Foss) were measured. Staphylococcal count was correlated to SCC (r=0.40), TBC (r=0.51), and SPC (r=0.53). Coliform count was correlated to TBC (r=0.33), but not to any of the other variables. Staphylococcus aureus did not correlate to SCC. The contribution of the staphylococcal count to the SPC was 31%, whereas the coliform count comprised only 1% of the SPC. The agreement of the repeated measurements was low. This study indicates that staphylococci in goat bulk milk are related to SCC and make a significant contribution to SPC. Because of the high variation in bacterial counts, repeated sampling is necessary to draw valid conclusions from bulk milk culturing. 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Star counts in M15 on U, B and V plates

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Calvani, M [Padua Univ. (Italy). Ist. di Astronomia; Nobili, L [Padua Univ. (Italy). Ist. di Fisica; Turolla, R [Scuola Internazionale Superiore di Studi Avanzati, Trieste (Italy)

1980-11-01

We present new counts of stars in M15, using plates in B, V and U. We are able to explore relatively close to the central parts of the cluster (0.1 pc) and we derive the best fitting parameters for the star distribution.

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

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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.

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

Science.gov (United States)

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.

Monitoring of airborne bacteria and aerosols in different wards of hospitals - Particle counting usefulness in investigation of airborne bacteria.

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Mirhoseini, Seyed Hamed; Nikaeen, Mahnaz; Khanahmd, Hossein; Hatamzadeh, Maryam; Hassanzadeh, Akbar

2015-01-01

The presence of airborne bacteria in hospital environments is of great concern because of their potential role as a source of hospital-acquired infections (HAI). The aim of this study was the determination and comparison of the concentration of airborne bacteria in different wards of four educational hospitals, and evaluation of whether particle counting could be predictive of airborne bacterial concentration in different wards of a hospital. The study was performed in an operating theatre (OT), intensive care unit (ICU), surgery ward (SW) and internal medicine (IM) ward of four educational hospitals in Isfahan, Iran. A total of 80 samples were analyzed for the presence of airborne bacteria and particle levels. The average level of bacteria ranged from 75-1194 CFU/m (3) . Mean particle levels were higher than class 100,000 cleanrooms in all wards. A significant correlation was observed between the numbers of 1-5 µm particles and levels of airborne bacteria in operating theatres and ICUs. The results showed that factors which may influence the airborne bacterial level in hospital environments should be properly managed to minimize the risk of HAIs especially in operating theaters. Microbial air contamination of hospital settings should be performed by the monitoring of airborne bacteria, but particle counting could be considered as a good operative method for the continuous monitoring of air quality in operating theaters and ICUs where higher risks of infection are suspected.

Mimicking Seawater For Culturing Marine Bacteria

DEFF Research Database (Denmark)

Rygaard, Anita Mac; Sonnenschein, Eva; Gram, Lone

2015-01-01

Only about 1% of marine bacteria have been brought into culture using traditional techniques. The purpose of this study was to investigate if mimicking the natural bacterial environment can increase culturability.We used marine substrates containing defined algal polymers or gellan gum as solidif......Only about 1% of marine bacteria have been brought into culture using traditional techniques. The purpose of this study was to investigate if mimicking the natural bacterial environment can increase culturability.We used marine substrates containing defined algal polymers or gellan gum...... as solidifying agents, and enumerated bacteria from seawater and algal exudates. We tested if culturability could be influenced by addition of quorum sensing signals (AHLs). All plates were incubated at 15°C. Bacterial counts (CFU/g) from algal exudates from brown algae were highest on media containing algal...... polymers. In general, bacteria isolated from algal exudates preferred more rich media than bacteria isolated from seawater. Overall, culturability ranged from 0.01 to 0.8% as compared to total cell count. Substitution of agar with gellan gum increased the culturability of seawater bacteria approximately...

Bacteriocidal activity of sanitizers against Enterococcus faecium attached to stainless steel as determined by plate count and impedance methods.

Science.gov (United States)

Andrade, N J; Bridgeman, T A; Zottola, E A

1998-07-01

Enterococcus faecium attached to stainless steel chips (100 mm2) was treated with the following sanitizers: sodium hypochlorite, peracetic acid (PA), peracetic acid plus an organic acid (PAS), quaternary ammonium, organic acid, and anionic acid. The effectiveness of sanitizer solutions on planktonic cells (not attached) was evaluated by the Association of Official Analytical Chemists (AOAC) suspension test. The number of attached cells was determined by impedance measurement and plate count method after vortexing. The decimal reduction (DR) in numbers of the E. faecium population was determined for the three methods and was analyzed by analysis of variance (P plate count method after vortexing, and impedance measurement, respectively. Plate count and impedance methods showed a difference (P measurement was the best method to measure adherent cells. Impedance measurement required the development of a quadratic regression. The equation developed from 82 samples is as follows: log CFU/chip = 0.2385T2-0.96T + 9.35, r2 = 0.92, P plate count method after vortexing. These data suggest that impedance measurement is the method of choice when evaluating the number of bacterial cells adhered to a surface.

Action of Antimicrobial Copper on Bacteria and Fungi Isolated from Commercial Poultry Hatcheries

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RFR Depner

Full Text Available ABSTRACT Since 2008, when the US Environmental Protection Agency (EPA registered copper and its alloys as an antimicrobial agent for contact surfaces, research has demonstrated their antimicrobial activity. The aim of this study was to evaluate the efficacy of antimicrobial copper against bacteria and fungi isolated from commercial poultry hatcheries in order to develop a microbiological control alternative in these environments. Samples were collected from the surfaces of hatcher baskets from two hatcheries. Mesophilic microorganisms and fungi/yeasts were isolated and standardized in concentration of 105 cells/mL. Four copper plates and four stainless steel plates were completely immersed for one minute in bacteria and fungi/yeasts solutions and left to dry for a day at room temperature. Subsequently, samples were collected from the metal plates with the aid of sterile swab and delimiter. These samples were planted onto Plate Count Agar (for mesophilic culture and Sabouraud Dextrose Agar (for fungi and yeast culture and incubated at 36°C for 48 hours and at 25°C for 5-7 days, respectively. After incubation, the colonies recovered from the plates were counted according to IN 62 of the Brazilian Ministry of Agriculture. Almost all contamination was eliminated from the surface of copper plates in a single day, while the stainless steel plates proved to be innocuous to the screened microorganisms. Copper, as a contact surface, proved to have important antimicrobial action on bacteria, fungi and yeasts common to hatcheries.

Total mesophilic counts underestimate in many cases the contamination levels of psychrotrophic lactic acid bacteria (LAB) in chilled-stored food products at the end of their shelf-life.

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Pothakos, Vasileios; Samapundo, Simbarashe; Devlieghere, Frank

2012-12-01

The major objective of this study was to determine the role of psychrotrophic lactic acid bacteria (LAB) in spoilage-associated phenomena at the end of the shelf-life of 86 various packaged (air, vacuum, modified-atmosphere) chilled-stored retail food products. The current microbiological standards, which are largely based on the total viable mesophilic counts lack discriminatory capacity to detect psychrotrophic LAB. A comparison between the total viable counts on plates incubated at 30 °C (representing the mesophiles) and at 22 °C (indicating the psychrotrophs) for 86 food samples covering a wide range - ready-to-eat vegetable salads, fresh raw meat, cooked meat products and composite food - showed that a consistent underestimation of the microbial load occurs when the total aerobic mesophilic counts are used as a shelf-life parameter. In 38% of the samples, the psychrotrophic counts had significantly higher values (+0.5-3 log CFU/g) than the corresponding total aerobic mesophilic counts. A total of 154 lactic acid bacteria, which were unable to proliferate at 30 °C were isolated. In addition, a further 43 with a poor recovery at this temperature were also isolated. This study highlights the potential fallacy of the total aerobic mesophilic count as a reference shelf-life parameter for chilled food products as it can often underestimate the contamination levels at the end of the shelf-life. Copyright © 2012 Elsevier Ltd. All rights reserved.

Petrifilm rapid S. aureus Count Plate method for rapid enumeration of Staphylococcus aureus in selected foods: collaborative study.

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Silbernagel, K M; Lindberg, K G

2001-01-01

A rehydratable dry-film plating method for Staphylococcus aureus in foods, the 3M Petrifilm Rapid S. aureus Count Plate method, was compared with AOAC Official Method 975.55 (Staphylococcus aureus in Foods). Nine foods-instant nonfat dried milk, dry seasoned vegetable coating, frozen hash browns, frozen cooked chicken patty, frozen ground raw pork, shredded cheddar cheese, fresh green beans, pasta filled with beef and cheese, and egg custard-were analyzed for S. aureus by 13 collaborating laboratories. For each food tested, the collaborators received 8 blind test samples consisting of a control sample and 3 levels of inoculated test sample, each in duplicate. The mean log counts for the methods were comparable for pasta filled with beef and cheese; frozen hash browns; cooked chicken patty; egg custard; frozen ground raw pork; and instant nonfat dried milk. The repeatability and reproducibility variances of the Petrifilm Rapid S. aureus Count Plate method were similar to those of the standard method.

Disinfection of bacteria attached to granular activated carbon.

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LeChevallier, M W; Hassenauer, T S; Camper, A K; McFeters, G A

1984-01-01

Heterotrophic plate count bacteria, coliform organisms, and pathogenic microorganisms attached to granular activated carbon particles were examined for their susceptibility to chlorine disinfection. When these bacteria were grown on carbon particles and then disinfected with 2.0 mg of chlorine per liter (1.4 to 1.6 mg of free chlorine residual per liter after 1 h) for 1 h, no significant decrease in viable counts was observed. Washed cells attached to the surface of granular activated carbon particles showed similar resistance to chlorine, but a progressive increase in sublethal injury was found. Observations made by scanning electron microscope indicated that granular activated carbon was colonized by bacteria which grow in cracks and crevices and are coated by an extracellular slime layer. These data suggest a possible mechanism by which treatment and disinfection barriers can be penetrated and pathogenic bacteria may enter drinking water supplies. Images PMID:6508306

IDENTIFICATION OF BACTERIA CAUSING DIARRHOEA IN HIV/AIDS PATIENTS AND ITS CORRELATION WITH CD4 COUNT

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Anand Premanand

2016-05-01

Full Text Available BACKGROUND The number of HIV-positive patients is increasing in India. Data on the prevalence of diarrhoea and the spectrum of bacteria responsible for diarrhoea in HIV- positive patients is lacking in our area. The identification of enteric pathogens in patients with HIV/AIDS is important because an increasing array of therapeutic regimens is becoming available to treat many of these infections. Thus, an attempt is done to elucidate the associations between causative bacteria of acute and chronic diarrhoea and CD4 count. METHODS Stool specimens were obtained over a period of eighteen months from HIV infected adults with diarrhoea presenting to Shri B M Patil Medical College Hospital and Research Centre, Vijayapura. In all patients with diarrhoea, stool specimens were examined by microscopy and cultures to identify bacterial pathogens and blood sample was analysed for CD4 count. RESULTS A total of 80 individuals were enrolled in this study. Cases included 46 males and 34 females. Among the cases, maximum subjects were found to be in the age group of 30-40 years in which 23 (62.2% were males and 14 (37.8% were females. 56 had acute and 24 had chronic diarrhoea. The percentages of bacteria isolated were 5 (8.9% in acute and 16 (66.7% in chronic diarrhoea respectively. The most common bacteria isolated was E. Coli (17.5% followed by Klebsiella (5% and Shigella Sps (3.75%. Patients with chronic diarrhoea had lower CD4 cell counts. The maximum bacterial isolation was in the patients whose CD4 cell counts were below 200 cells/mm3. CONCLUSION Bacterial isolation was most strongly associated with low CD4 counts and chronic diarrhoea. E. coli was isolated maximum among all the bacteria in the HIV patients. Over two-thirds of diarrhoeal episodes were undiagnosed, suggesting that unidentified agents or primary HIV enteropathy are important causes of diarrhoea in this population. There is a strong negative association between duration of diarrhoea and CD4

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.

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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.

The presence of embedded bacterial pure cultures in agar plates stimulate the culturability of soil bacteria

DEFF Research Database (Denmark)

Burmølle, Mette; Johnsen, Kaare; Abu Al-Soud, Waleed Mohamad Abdel F

2009-01-01

Traditional methods for bacterial cultivation recover only a small fraction of bacteria from all sorts of natural environments, and attempts have been made to improve the bacterial culturability. Here we describe the development of a cultivation method, based on the embedment of pure bacterial...... cultures in between two layers of agar. Plates containing either embedded Pseudomonas putida or Arthrobacter globiformis resulted in higher numbers of CFUs of soil bacteria (21% and 38%, respectively) after 833 h of incubation, compared to plates with no embedded strain. This indicates a stimulatory effect...... of the bacterial pure cultures on the cultivation of soil bacteria. Analysis of partial 16S rRNA gene sequences revealed a phylogenetical distribution of the soil isolates into 7 classes in 4 phyla. No difference was observed at the phylum or class level when comparing isolates grouped according to embedded strain...

Rapid diagnostics of the bacteria in air

Energy Technology Data Exchange (ETDEWEB)

Belov Nikolai, N. [ATECH KFT, Budapest (Hungary)

2000-07-01

Presence of the bacteria and viruses in the air is great problem now. Terrorists are going to use the bacteria weapon. Now biotechnology provides very cheap equipment ({approx} $500) for modification of the bacteria sorts. It may be used for receiving of new variants of the bacteriological weapon. And presence of one small bacteria aerosol generator in the international airport during several days will start the dangerous epidemic incidence the entire world. From another side - poor countries with hot and wet weather are continuously producing new and new dangerous bacteria. Every year epidemic waves of influence are going from China, India or Africa. And once up a time it will be epidemic explosive with fast lethal finish. Methods of estimation of the bio-aerosols in Air of City are very poor. Standard Bio-aerosol sampler has two conflicting demands. From one side the bio-sampler needs in great air volume of sample with great efficiency of separation of aerosol particles from measured air. From another side all selected particles needs in great care. This demand carried out from method of measurement of bacteria in sample by counting of colonies that grew from bacteria on nutrient media after incubation time. It is a problem to prevent bacterial flora from death during collecting aerosol sample. This time of growth and counting of colony is so long that result of this measurement will be unusable if it will be terrorist action of start of bacteriological was. Here presented new methods for fast diagnostics of the bacteria in the air. It consists from 4 general parts: (1) Micro-droplet method for diagnostics of biological active substances in aerosol sample. This method allows to control the bio-particle position on the plate, to use series of biochemistry species for analytical reaction for this small bio-particle. Small volume of biochemical reaction reduces noise. This method provides extremely high sensitivity for discovering of biological material. (2

Retreived bacteria from Noctiluca miliaris (green) bloom of the northeastern Arabian Sea

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Basu, Subhajit; Matondkar, S. G. Prabhu; Furtado, Irene

2013-01-01

In recent years, seasonal blooms of the dinoflagellate Noctiluca miliaris have appeared in the open-waters of the northern Arabian Sea (NAS). This study provides the first characterization of bacteria from a seasonal bloom of green Noctiluca of NAS (20°N-17°N and 64°E-70°E), during the spring-inter-monsoon cruise of Sagar Sampada 253, in March 2007. Bacterial growth as assessed by most-probable number (MPN) and plate counts, revealed `variable-physiotypes' over a wide range of salinities (0%-25% w/v NaCl), pH levels (5-8.5), and organic nutrient strengths, in comparison to non-bloom waters. MPN indices of bacteria in surface waters of bloom stations *DWK and *PRB, corresponded to (3.08-4.41)×103 cells/mL at 3.5% NaCl (w/v), and (2.82-9.49)×102 cells/mL at 25% (w/v) NaCl in tryptone-yeast extract broth (TYE). Plate counts were (1.12-4)×106 CFU/mL at 0% (w/v) NaCl, (1.28-3.9)×106 CFU/mL at 3.5% (w/v) NaCl, and (0.4-7)×104 CFU/mL at 25% NaCl (w/v) on TYE. One-tenth-strength Zobell's gave (0.6-3.74)×105 CFU/mL at pH 5 to (3.58-7.5)×105 CFU/mL at pH 8.5. These bacteria were identified to the genera Bacillus, Cellulomonas, Staphylococcus, Planococcus, Dietzia, Virgibacillus, Micrococcus, Sporosarcinae, Leucobacter, and Halomonas. The identity of three strains (GUFBSS253N2, GUFBSS253N30, and GUFBSS253N84) was confirmed through 16S rDNA sequence homology as Bacillus cohnii, Bacillus flexus, and Bacillus cereus. The ˜2-3-fold higher plate counts of culturable bacteria from the open-waters of the NAS indicate that these bacteria could critically determine the biogeochemical dynamics of the bloom and its milieu. The role of these bacteria in sustaining/terminating the bloom is under evaluation.

Determination of Complement-Mediated Killing of Bacteria by Viability Staining and Bioluminescence

OpenAIRE

Virta, Marko; Lineri, Sanna; Kankaanpää, Pasi; Karp, Matti; Peltonen, Karita; Nuutila, Jari; Lilius, Esa-Matti

1998-01-01

Complement-mediated killing of bacteria was monitored by flow cytometric, luminometric, and conventional plate counting methods. A flow cytometric determination of bacterial viability was carried out by using dual staining with a LIVE/DEAD BacLight bacterial viability kit. In addition to the viable cell population, several other populations emerged in the fluorescence histogram, and there was a dramatic decrease in the total cell count in the light-scattering histogram in the course of the co...

Do different standard plate counting (IDF/ISSO or AOAC) methods interfere in the conversion of individual bacteria counts to colony forming units in raw milk?

Science.gov (United States)

Cassoli, L D; Lima, W J F; Esguerra, J C; Da Silva, J; Machado, P F; Mourão, G B

2016-10-01

This study aimed to establish the correlation between individual bacterial count (IBC) obtained by flow cytometry and the number of colony forming units (CFU) determined by standard plate count (SPC) in raw milk using two different reference methodologies: the methodology of the International Dairy Federation (IDF) - International Organization for Standardization (ISO) 4833, incubation for 72 h at 30°C and the methodology of the Association of Official Analytical Chemists (AOAC), incubation for 48 h at 35°C. For this, 100 bovine milk samples (80 ml) from different farms were collected in a sterile bottle and maintained refrigerated at 4°C and were delivered to the laboratory. In the laboratory, the samples were divided into two vials of 40 ml each. Then, half of the vials were forwarded for the SPC analysis, and the other half were analysed using the equipment BactoScan FC. The analyses by flow cytometry and SPC were performed at the same time (maximum deviation of +/- 1 h). To transform the data from IBC ml(-1) to CFU ml(-1) (IDF or AOAC methodology), a standard linear regression equation was used, as recommended by IDF/ISO-196. The difference between the reference methodologies affects the equation that transforms IBC into CFU and therefore the accuracy of the results. The results estimated by the equation using the ISO 4833 methodology were on average 0·18 log units higher than the results estimated using the equation using the AOAC methodology. After the comparison of the methodologies, it was concluded that there is an impact of the reference methodologies on the conversion of the results from IBC to CFU. Depending on the methodology adopted by each laboratory or country, there may not be equivalence in the results. Hence, the laboratories specialized in milk quality analysis that have changed their methodology for analysis, passing from the MAPA (AOAC) methodology to the IDF standard, need to develop new conversion equations to make their

Effect of irradiation and storage post-irradiation of black pepper (Piper nigrum L.) on counts of microorganisms hygienic indicator using methods of conventional analysis and PETRIFILMTM plates

International Nuclear Information System (INIS)

Jaimes, Marcial Ibo Silva

1988-01-01

Fifteen samples of ground black pepper (Piper nigrum L.) purchased in Sao Paulo local stores, were submitted to irradiation in doses of 3, 6 and 10 kGy. All irradiated samples, including non-irradiated controls, were submitted to counts of yeasts and molds, aerobes (APC), coliforms and mesophilic aerobic spore formers (MASC), using conventional plate count methods and PETRIFILM TM plates. For yeasts and molds count, acidified potato dextrose agar (PDA) an PETRIFILM TM PFYM plates were used. For aerobes, plate count agar (PCA) and PETRIFILM TM PFAC plates were used. Violet red bile agar (VRBA) and PETRIFILM TM PFEC plates were employed for enumeration of coliforms. Counts of these groups of microorganisms obtained through the traditional plating procedures did not differ significantly from those using the corresponding PETRIFILM TM plates. In samples submitted to irradiation, a dose of 10 kGy caused a decrease of the yeasts and molds count from 10 4 -10 5 to less than 10 cfu/g. The same dose caused a decrease of the aerobic counts from 10 7 -10 8 to 10 2 -10 3 cfu/g, of coliforms from 10 4 -10 5 to less than 10 cfu/g and MASC from 10 6 -10 7 cfu/g to 10-10 2 cfu/g. The introduction of a injury repair step in the counting procedure resulted in a 32 to 89% increase in the number of coliforms. However, this additional step did not improve significantly the counts of MASC. After 270 days of storage of samples irradiated with 3 kGy, a decrease in the yeasts and molds population from 10 3 to 20 cfu/g was observed. The APC population in these samples was reduced from 5,0x10 6 to 2,4x10 4 cfu/g; in those irradiated with 6 kGy the reduction was from 4,0x10 4 to 5,0x10 3 cfu/g and in those irradiated with 10 kGy the counts were reduced from 30 to less than 10 cfu/g. After the same time of storage, the coliform population in non irradiated samples decreased from 2,8x10 5 to 1,5x10 4 cfu/g and from 9,1x10 3 to 20 cfu/g in those irradiated with 3 kGy. Similarly, the MASC

Influence of Heat Shock Temperatures and Fast Freezing on Viability of Probiotic Sporeformers and the Issue of Spore Plate Count Versus True Numbers

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Mojtaba Jafari

2016-02-01

Full Text Available Background and Objectives: The purpose of the present study was to investigate effects of various heat shock conditions and fast freezing and subsequent thawing on the viability and recovery of Bacillus coagulans and Bacillus subtilis as probiotic sporeformers, and also to compare spore plate and microscopic counts. Materials and Methods: After preparing the final suspensions of B. coagulans and Bacillus subtilis subsp. Natto spores, they were spread-plated before and after fast freezing treatment (-70°C for about 1 min. Heat shock treatments of the spores were carried out at 68oC for 15, 20, and 30 min as well as at 80oC for 10 and 15 min. Concentrations of the examined probiotic sporeformers were determined simultaneously by plate enumerations and microscopically determined counts. Student’s t-test and one-way analysis of variance (ANOVA of SPSS were used for statistical analysis of the data. Analysis of DoE results was carried out using Minitab. Results: The results presented here show that the highest recovery rates for B. coagulans (14.75 log CFU/mL and B. subtilis spores (14.80 log CFU/mL were under a heat shock condition of 68°C for 20 min in nutrient agar (p<0.05. In addition, the survival rates of B. coagulans and B. subtilis spores under the fast freezing and subsequent thawing condition were about 90% and 88%, respectively. Plate counts differed significantly from counts determined microscopically, with differences of almost 0.5 and 0.8 log for B. coagulans and B. subtilis spores, respectively (p<0.05. In addition, DoE results of the study revealed that both factors of spore count method and only freezing factor in fast freezing treatment have a significant effect on concentrations of the spores examined (p<0.05. Conclusions: Heat shock conditions, freezing and subsequent thawing circumstances, and plate counts or enumerations determined microscopically have significant influences on the viability of probiotic sporeformers and

Evaluation of Petrifilm™ Select E. coli Count Plate medium to discriminate antimicrobial resistant Escherichia coli

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Jensen Lars

2008-09-01

Full Text Available Abstract Background Screening and enumeration of antimicrobial resistant Escherichia coli directly from samples is needed to identify emerging resistant clones and obtain quantitative data for risk assessment. Aim of this study was to evaluate the performance of 3M™ Petrifilm™ Select E. coli Count Plate (SEC plate supplemented with antimicrobials to discriminate antimicrobial-resistant and non-resistant E. coli. Method A range of E. coli isolates were tested by agar dilution method comparing the Minimal Inhibitory Concentration (MIC for eight antimicrobials obtained by Mueller-Hinton II agar, MacConkey agar and SEC plates. Kappa statistics was used to assess the levels of agreement when classifying strains as resistant, intermediate or susceptible. Results SEC plate showed that 74% of all strains agreed within ± 1 log2 dilution when comparing MICs with Mueller-Hinton II media. High agreement levels were found for gentamicin, ampicillin, chloramphenicol and cefotaxime, resulting in a kappa value of 0.9 and 100% agreement within ± 1 log2 dilution. Significant variances were observed for oxytetracycline and sulphamethoxazole. Further tests showed that the observed discrepancy in classification of susceptibility to oxytetracycline by the two media could be overcome when a plate-dependent breakpoint of 64 mg/L was used for SEC plates. For sulphamethoxazole, SEC plates provided unacceptably high MICs. Conclusion SEC plates showed good agreement with Mueller-Hinton II agar in MIC studies and can be used to screen and discriminate resistant E. coli for ampicillin, cephalothin, streptomycin, chloramphenicol, cefotaxime and gentamicin using CLSI standardized breakpoints, but not for sulphamethoxazole. SEC plates can also be used to discriminate oxytetracycline-resistant E. coli if a plate-dependent breakpoint value of 64 mg/L is used.

Detection of irradiated food by using direct epifluorescent filter technique/aerobic plate count method (DEFT/APC)

International Nuclear Information System (INIS)

Zhao Yongfu; Li Lili; Wang Changbao; Ji Ping; Wang Chao; Wang Zhidong

2010-01-01

The Direct Epifluorescent Filter Technique/Aerobic Plate Count technique(DEFT/APC) can be used to identify the irradiated food by comparing the DEFT and APC counts of the samples prior to the irradiation and after. This technology was tested by using spice and dried marine fish as testing materials in this study. The results shows that the index, log (DEFT/APC) > 4.0, can indicate that the samples have been irradiated at a dose level higher than 1.0 kGy. It also indicates that the detecting sensitivity was affected by the initial value of APC and D 10 value of the samples. (authors)

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

Science.gov (United States)

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.

The Application of the Photographic Plate to the Quantitative Determination of Activities by Track Counts

International Nuclear Information System (INIS)

Broda, E.

1946-01-01

This report was written by E. Broda at the Cavendish Laboratory (Cambridge) in August 1946 and is about the application of the photographic plate to the quantitative determination of activities by track counts. This report includes the experiment description and the discussion of the results and consists of 4 parts: 1) Introduction 2) Estimation of Concentrations 3) The uptake of U in different conditions 4) The upper limits of the fission Cross sections of Bi and Pb. (nowak)

The Application of the Photographic Plate to the Quantitative Determination of Activities by Track Counts

Energy Technology Data Exchange (ETDEWEB)

Broda, E.

1946-07-01

This report was written by E. Broda at the Cavendish Laboratory (Cambridge) in August 1946 and is about the application of the photographic plate to the quantitative determination of activities by track counts. This report includes the experiment description and the discussion of the results and consists of 4 parts: 1) Introduction 2) Estimation of Concentrations 3) The uptake of U in different conditions 4) The upper limits of the fission Cross sections of Bi and Pb. (nowak)

Pathogenic Assay of Probiotic Bacteria Producing Proteolytic Enzymes as Bioremediation Bacteria Against Vannamei Shrimp Larvae (Litopenaeus vannamei

Directory of Open Access Journals (Sweden)

Wilis Ari Setyati

2017-06-01

Full Text Available Application of bacteria in bioremediation of shrimp culture ponds is one of the methods used to clean internal pollutants. This study aimed to evaluate the pathogenicity of extracellular proteolytic enzyme produced by the probiotic bacteria as bioremediation bacteria on vannamei shrimp larvae culture. There were five probiotic bacteria, which were successfully isolated from the sediments served as substrate in mangrove area. The isolated bacteria were coded in number as 13, 19, 30, 33, and 36. Pathogenic bacteria Vibrio harveyi was used as positive control. Pathogenic assay was carried out in two different bacterial concentrations, i.e. 10⁸ and 10⁶ cells.mL-1. The results showed that the lowest survival rate (SR of shrimp larvae in positive control V. harveyi was 53 and 65%. Whereas isolates with the highest SR value (100% were obtained from bacteria coded as 13 and 30. Isolates no. 19, 33 and 36 had SR of more than 90%. Total plate count (TPC data showed that the bacteria increased significantly at the end of the study with an average increase value of 24%. The smallest TPC value was shown by bacterial isolate no. 19, while the largest was obtained from the isolate no. 13. These results suggest that all probiotic bacteria were not pathogenic to the vannamei shrimp larvae.   Keywords: aquaculture, shrimp, bioremediation, pathogenesis, vibrio.

Differential carbohydrate media and anaerobic replica plating techniques in delineating carbohydrate-utilizing subgroups in rumen bacterial populations.

OpenAIRE

Leedle, J A; Hespell, R B

1980-01-01

A basal (BC) medium devoid of added carbohydrates, a complete (CC) medium containing nine carbohydrates were developed for enumerating rumen bacteria. The colony counts on the BC medium were 85 to 100% of those obtained on the CC medium. These colonies were pinpoint size (less than or equal to mm in diameter) but increased in size (2 to 5 mm in diameter) when carbohydrates were subsequently added. With the CC medium or other media tested, the colony counts were 20 to 50% higher on plates than...

Salivary cariogenic bacteria counts are associated with obesity in student women at a Malaysian university.

Science.gov (United States)

Yeo, Wey-Zheng; Lim, Sheng-Pei; Say, Yee-How

2018-01-01

The counts of cariogenic bacteria lactobacilli and mutans streptococci have been studied and correlated with sugar intake. This study was to investigate the association between salivary lactobacilli and mutans streptococci counts with sweet food eating behavior and sweet sensitivity among 120 Malaysian women (101 ethnic Chinese, 19 ethnic Indians), while taking into account anthropometric and menstruation variables. Demographics, anthropometric measurements and menstrual history were taken. Hedonic preference, intake frequency of a list of sweet foods, intensity perception and pleasantness ratings of sweet stimuli were assessed. Saliva was collected for lactobacilli and mutans streptococci culture. We found that centrally obese subjects (high waist circumference and waist-hip ratio) had significantly higher salivary lactobacilli and mutans streptococci counts (all pMalaysian women are associated with central obesity, but not sweet food eating behaviour, sweet sensitivity and menstruation variables. Salivary microbiome analysis could be useful as a potential diagnostic indicator of diseases such as obesity.

Disposable bioluminescence-based biosensor for detection of bacterial count in food.

Science.gov (United States)

Luo, Jinping; Liu, Xiaohong; Tian, Qing; Yue, Weiwei; Zeng, Jing; Chen, Guangquan; Cai, Xinxia

2009-11-01

A biosensor for rapid detection of bacterial count based on adenosine 5'-triphosphate (ATP) bioluminescence has been developed. The biosensor is composed of a key sensitive element and a photomultiplier tube used as a detector element. The disposable sensitive element consists of a sampler, a cartridge where intracellular ATP is chemically extracted from bacteria, and a microtube where the extracted ATP reacts with the luciferin-luciferase reagent to produce bioluminescence. The bioluminescence signal is transformed into relevant electrical signal by the detector and further measured with a homemade luminometer. Parameters affecting the amount of the extracted ATP, including the types of ATP extractants, the concentrations of ATP extractant, and the relevant neutralizing reagent, were optimized. Under the optimal experimental conditions, the biosensor showed a linear response to standard bacteria in a concentration range from 10(3) to 10(8) colony-forming units (CFU) per milliliter with a correlation coefficient of 0.925 (n=22) within 5min. Moreover, the bacterial count of real food samples obtained by the biosensor correlated well with those by the conventional plate count method. The proposed biosensor, with characteristics of low cost, easy operation, and fast response, provides potential application to rapid evaluation of bacterial contamination in the food industry, environment monitoring, and other fields.

Assessment of the efficiency of SimPlate™ total plate count color indicator (TPC CI to quantify mesophilic aerobic microorganisms in pasteurized milk Avaliação da eficiência do SimPlate™ Total Plate Count Color Indicator (TPC CI para enumeração de microrganismos aeróbios mesófilos em leite pasteurizado

Directory of Open Access Journals (Sweden)

Luís Augusto Nero

2002-01-01

Full Text Available The SimPlate™ TPC CI system is a rapid method to count mesophilic aerobic microorganisms (MAM in foods, based on the use of resazurine to indicate bacterial growth. Its efficiency in pasteurized milk was evaluated using 142 pasteurized milk samples (38 type A, 43 type B and 61 type C collected in Londrina, PR. The standard plating method, using Plate Count Agar (PCA was used for comparison. The plates of both systems were incubated at 35ºC and read after 24h and 48h. The occurrence of false-positive and false-negative wells and the predominant microorganisms in them were also evaluated. The results were compared by simple correlation and mean variance analyses. The correlation (r and mean variance values were 0.6811 and 0.7583 for the results obtained after 24h, respectively, and 0.9126 and 0.0842 for the results obtained after 48h, respectively. These results indicate that the performance of the system increases when the plates are incubated for 48h. When the three types of milk were evaluated separately, these values were 0.9285 and 0.0817 for type A milk, 0.9231 and 0.0466 for type B milk and 0.7209 and 0.1082 for type C milk. These results indicate that the better the quality of the milk the better the performance of SimPlate™ TPC CI. False-negative wells, found more frequently in samples with high MAM counts, were caused by Gram positive microorganisms, poorly detected by the SimPlate™ TPC CI system because they grew slowly and had low reduction capacity. The results indicated a higher efficiency of the SimPlate™ TPC CI system in the reading at 48h.O sistema SimPlate™ TPC CI é um método rápido para enumeração de microrganismos aeróbios mesófilos (MAM em alimentos que utiliza a resazurina como substância indicadora de crescimento bacteriano. Para avaliar sua eficiência em leite pasteurizado, 142 amostras (38 de leite tipo A, 43 de leite tipo B e 61 de leite tipo C foram colhidas em Londrina, PR, e analisadas pelo SimPlate

Spark counting technique with an aluminium oxide film

International Nuclear Information System (INIS)

Kawai, H.; Koga, T.; Morishima, H.; Niwa, T.; Nishiwaki, Y.

1980-01-01

Automatic spark counting of etch-pits on a polycarbonate film produced by nuclear fission fragments is now used for neutron monitoring in several countries. A method was developed using an aluminium oxide film instead of a polycarbonate as the neutron detector. Aluminium oxide films were prepared as follows: A cleaned aluminium plate as an anode and a nickel plate as a cathode were immersed in dilute sulfuric acid solution and electric current flowed between the electrodes at 12degC for 10-30 minutes. Electric current density was about 10 mA/cm 2 . The aluminium plate was then kept in boiling water for 10-30 minutes for sealing. The thickness of the aluminium oxide layer formed was about 1μm. The aluminium plate attached to a plate of suitable fissionable material, such as uranium or thorium, was irradiated with neutrons and set in a usual spark counter for fission track counting. One electrode was the aluminium plate and the other was an aluminized polyester sheet. Sparked pulses were counted with a usual scaler. The advantage of using spark counting with an aluminium oxide film for neutron monitoring is rapid measurement of neutron exposure, since chemical etching which is indispensable for spark counting with a polycarbonate detector film, is not needed. (H.K.)

Isolation and characterization of aerobic culturable arsenic-resistant bacteria from surfacewater and groundwater of Rautahat District, Nepal.

Science.gov (United States)

Shakya, S; Pradhan, B; Smith, L; Shrestha, J; Tuladhar, S

2012-03-01

Arsenic (As) contamination of groundwater is a serious Environmental Health Management issue of drinking water sources especially in Terai region of Nepal. Many studies have reported that due to natural abundance of arsenic in the environment, various bacteria have developed different resistance mechanisms for arsenic compound. In this study, the culturable arsenic-resistant bacteria indigenous to surfacewater as well as groundwater from Rautahat District of Nepal were randomly isolated by standard plate count method on the basis of viable growth on plate count agar amended with arsenate ranging from 0, 0.5, 10, 40, 80 to 160 milligram per liter (mg/l). With respect to the morphological and biochemical tests, nine morphologically distinct potent arsenate tolerant bacteria showed relatedness with Micrococcus varians, Micrococcus roseus, Micrococcus luteus, Pseudomonas maltophilia, Pseudomonas sp., Vibrio parahaemolyticus, Bacillus cereus, Bacillus smithii 1 and Bacillus smithii 2. The isolates were capable of tolerating more than 1000 mg/l of arsenate and 749 mg/l of arsenite. Likewise, bioaccumulation capability was highest with M. roseus (85.61%) and the least with B. smithii (47.88%) indicating the potential of the organisms in arsenic resistance and most probably in bioremediation. Copyright © 2011 Elsevier Ltd. All rights reserved.

Characterization of the spoilage lactic acid bacteria in “sliced vacuum-packed cooked ham”

Science.gov (United States)

Kalschne, Daneysa Lahis; Womer, Rute; Mattana, Ademir; Sarmento, Cleonice Mendes Pereira; Colla, Luciane Maria; Colla, Eliane

2015-01-01

The lactic acid bacteria are involved with food fermentation and in such cases with food spoilage. Considering the need to reduce the lactic acid bacteria growth in meat products, the aim of this work was to enumerated and investigated the lactic acid bacteria present on sliced vacuum-packed cooked ham stored at 4 °C and 8 °C for 45 days by phenotypic and molecular techniques. The quantification showed that the lactic acid bacteria were present from the first day with mean count of 1.98 log cfu/g for the four batches analyzed. The lactic acid bacteria grew rapidly on the samples, and plate counts around 7.59 log cfu/g and 8.25 log cfu/g were detected after 45 days of storage at 4 °C and 8 °C, respectively; storage temperatures studied showed significant influence on the microorganism in study growth. The predominant lactic acid bacteria associated with the spoilage samples at one day of storage includes Lactobacillus sp., the phenotypic overlap Leuconostoc / Weissella sp. and Enterococcus sp. At 45 days of storage at 4 and 8 °C the mainly specie was Lactobacillus curvatus , following by Lactobacillus sakei and Leuconostoc mesentereoides ; the Enterococcus sp. was not present in the samples. PMID:26221105

Determination of Bacterial Growth in Culture Media

International Nuclear Information System (INIS)

Elly Ellyna Rashid; Shariza Hanim Zainal Abidin; Mok, P.S.

2015-01-01

Bacteria is one of the important microorganism in our daily life. Bacteria provides human beings with products in the field of medical, industry, food, agriculture and others. Determination of bacteria growth is important so that we can enjoy the most benefit from it. Spread-plate method is one of the methods to obtain the bacterial counts. Agar plates, such as Nutrient Agar or Plate Count Agar are usually used for this purpose. Bacterial culture will be diluted first before being spread on the agar plate and incubated at specific temperature. The number of bacteria in colony-forming unit (CFU) will be counted the next day. The count will be used to determine the bacterial growth. (author)

Evaluation of Petrifilm™ method for enumerating aerobic bacteria in Crottin goat's cheese Evaluación de PetrifilmTM para la enumeración de bacterias aerobias en queso de cabra Crottin

Directory of Open Access Journals (Sweden)

G.B. de Sousa

2005-12-01

Full Text Available The PetrifilmTM Aerobic Count Plate (ACP developed by 3M laboratories, is a ready-to-use culture medium system, useful for the enumeration of aerobic bacteria in food. PetrifilmTMwas compared with a standard method in several different food products with satisfactory results. However, many studies showed that bacterial counts in PetrifilmTM were significantly lower than those obtained with conventional methods in fermented food. The purpose of this study was to compare the PetrifilmTM method for enumerating aerobic bacteria with a conventional method (PCA in Crottin goat's cheese. Thirty samples were used for the colony count. The mean count and standard deviation were 7.18 ± 1.17 log CFU g-1 on PCA and 7.11 ± 1.05 log CFU g-1 on PetrifilmTM. Analysis of variance revealed no significant differences between both methods (t = 1.33, P = 0.193. The Pearson correlation coefficient (0.971, P=0.0001 indicated a strong linear relationship between the PetrifilmTM and the standard method. The results showed that PetrifilmTM is suitable and a convenient alternative to this standard method for the enumeration of aerobic flora in goat soft cheese.PetrifilmTM Aerobic Count Plate (ACP desarrollado por 3M es un sistema listo para usar, empleado para el recuento de bacterias aerobias en alimentos. PetrifilmTMfue comparado con los métodos estándar en diferentes productos alimenticios con resultados satisfactorios. Sin embargo, en alimentos fermentados, algunos estudios mostraron que el recuento de bacterias aerobias en PetrifilmTM fue significativamente menor que aquellos obtenidos con los métodos convencionales (PCA. El propósito de este estudio fue comparar el método PetrifilmTM para el recuento de bacterias aerobias con un método convencional en queso de cabra Crottin. Se usaron 30 muestras para el recuento de colonias. Las medias y desviaciones estándar fueron 7,18 ± 1,17 log UFC g-1 en PCA y 7,11 ± 1,05 log UFC g-1 en PetrifilmTM. El análisis de

Microbiological diversity and prevalence of spoilage and pathogenic bacteria in commercial fermented alcoholic beverages (beer, fruit wine, refined rice wine, and yakju).

Science.gov (United States)

Jeon, Se Hui; Kim, Nam Hee; Shim, Moon Bo; Jeon, Young Wook; Ahn, Ji Hye; Lee, Soon Ho; Hwang, In Gyun; Rhee, Min Suk

2015-04-01

The present study examined 469 commercially available fermented alcoholic beverages (FABs), including beer (draft, microbrewed, and pasteurized), fruit wine (grape and others), refined rice wine, and yakju (raw and pasteurized). Samples were screened for Escherichia coli and eight foodborne pathogens (Bacillus cereus, Campylobacter jejuni, Clostridium perfringens, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus, and Yersinia enterocolitica), and the aerobic plate count, lactic acid bacteria, acetic acid bacteria, fungi, and total coliforms were also enumerated. Microbrewed beer contained the highest number of microorganisms (average aerobic plate count, 3.5; lactic acid bacteria, 2.1; acetic acid bacteria, 2.0; and fungi, 3.6 log CFU/ml), followed by draft beer and yakju (P beer samples) and B. cereus (detected in all FABs) were present in some products. B. cereus was detected most frequently in microbrewed beer (54.8% of samples) and nonpasteurized yakju (50.0%), followed by pasteurized yakju (28.8%), refined rice wine (25.0%), other fruit wines (12.3%), grape wine (8.6%), draft beer (5.6%), and pasteurized beer (2.2%) (P < 0.05). The finding that spore-forming B. cereus and coliform bacteria can survive the harsh conditions present in alcoholic beverages should be taken into account (alongside traditional quality indicators such as the presence of lactic acid-producing bacteria, acetic acid-producing bacteria, or both) when developing manufacturing systems and methods to prolong the shelf life of high-quality FAB products. New strategic quality management plans for various FABs are needed.

Effects of hot water application after defeathering on the levels of Campylobacter, coliform bacteria, and Escherichia coli on broiler carcasses.

Science.gov (United States)

Berrang, M E; Dickens, J A; Musgrove, M T

2000-11-01

Scalding has been found to lower the levels of Campylobacter on broiler carcasses. However, the numbers recovered from whole-carcass rinse samples increase following defeathering. This study was undertaken to examine the effect of a second scald applied after defeathering on microbial levels recovered from carcass rinses. Four treatments were evaluated: 1) immersion at 60 C for 28 s 30 min after defeathering, 2) immersion at 60 C for 28 s immediately after defeathering, 3) spray at 73 C for 20 s 30 min after defeathering, and 4) spray at 71 C for 20 s immediately after defeathering. As reported earlier, a significant increase in Campylobacter counts per mL whole carcass rinse was noted after carcasses were defeathered. However, when applied 30 min after defeathering, neither the immersion nor the spray second scald treatments lowered the Campylobacter counts. Likewise, neither treatment had any affect on Escherichia coli or coliform bacteria counts, even though total counts were slightly reduced by the treatments. When the second scald treatment immediately followed defeathering, the same trends were observed. Campylobacter counts after the second scald remained at the postpick levels, as did counts for E. coli and coliform bacteria, but total plate counts were slightly reduced. Overall, it would appear that a postscald treatment gentle enough not to alter the carcass appearance or meat quality would not effectively lower Campylobacter, E. coli, or coliform bacteria counts.

Rapid and reliable identification of Gram-negative bacteria and Gram-positive cocci by deposition of bacteria harvested from blood cultures onto the MALDI-TOF plate.

Science.gov (United States)

Barnini, Simona; Ghelardi, Emilia; Brucculeri, Veronica; Morici, Paola; Lupetti, Antonella

2015-06-18

Rapid identification of the causative agent(s) of bloodstream infections using the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) methodology can lead to increased empirical antimicrobial therapy appropriateness. Herein, we aimed at establishing an easier and simpler method, further referred to as the direct method, using bacteria harvested by serum separator tubes from positive blood cultures and placed onto the polished steel target plate for rapid identification by MALDI-TOF. The results by the direct method were compared with those obtained by MALDI-TOF on bacteria isolated on solid media. Identification of Gram-negative bacilli was 100 % concordant using the direct method or MALDI-TOF on isolated bacteria (96 % with score > 2.0). These two methods were 90 % concordant on Gram-positive cocci (32 % with score > 2.0). Identification by the SepsiTyper method of Gram-positive cocci gave concordant results with MALDI-TOF on isolated bacteria in 87 % of cases (37 % with score > 2.0). The direct method herein developed allows rapid identification (within 30 min) of Gram-negative bacteria and Gram-positive cocci from positive blood cultures and can be used to rapidly report reliable and accurate results, without requiring skilled personnel or the use of expensive kits.

Rapid and reliable identification of Gram-negative bacteria and Gram-positive cocci by deposition of bacteria harvested from blood cultures onto the MALDI-TOF plate.

OpenAIRE

Barnini, S; Ghelardi, Emilia; Brucculeri, V; Morici, Paola; Lupetti, Antonella

2015-01-01

Background Rapid identification of the causative agent(s) of bloodstream infections using the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) methodology can lead to increased empirical antimicrobial therapy appropriateness. Herein, we aimed at establishing an easier and simpler method, further referred to as the direct method, using bacteria harvested by serum separator tubes from positive blood cultures and placed onto the polished steel target plate for rapid identif...

Analytic Method on Characteristic Parameters of Bacteria in Water by Multiwavelength Transmission Spectroscopy

Directory of Open Access Journals (Sweden)

Yuxia Hu

2017-01-01

Full Text Available An analytic method together with the Mie scattering theory and Beer-Lambert law is proposed for the characteristic parameter determination of bacterial cells (Escherichia coli 10389 from multiwavelength transmission spectroscopy measurements. We calculate the structural parameters of E. coli cells, and compared with the microscopy, the relative error of cell volume is 7.90%, the cell number is compared with those obtained by plate counting, the relative error is l.02%, and the nucleic content and protein content of single E. coli cells are consistent with the data reported elsewhere. The proposed method can obtain characteristic parameters of bacteria as an excellent candidate for the rapid detection and identification of bacteria in the water.

Rapid quantification of live/dead lactic acid bacteria in probiotic products using high-sensitivity flow cytometry

Science.gov (United States)

He, Shengbin; Hong, Xinyi; Huang, Tianxun; Zhang, Wenqiang; Zhou, Yingxing; Wu, Lina; Yan, Xiaomei

2017-06-01

A laboratory-built high-sensitivity flow cytometer (HSFCM) was employed for the rapid and accurate detection of lactic acid bacteria (LAB) and their viability in probiotic products. LAB were stained with both the cell membrane-permeable SYTO 9 green-fluorescent nucleic acid stain and the red-fluorescent nucleic acid stain, propidium iodide, which penetrates only bacteria with compromised membranes. The side scatter and dual-color fluorescence signals of single bacteria were detected simultaneously by the HSFCM. Ultra-high temperature processing milk and skim milk spiked with Lactobacillus casei were used as the model systems for the optimization of sample pretreatment and staining. The viable LAB counts measured by the HSFCM were in good agreement with those of the plate count method, and the measured ratios between the live and dead LAB matched well with the theoretical ratios. The established method was successfully applied to the rapid quantification of live/dead LAB in yogurts and fermented milk beverages of different brands. Moreover, the concentration and viability status of LAB in ambient yogurt, a relatively new yet popular milk product in China, are also reported.

Microbiological assessment of house and imported bottled water by comparison of bacterial endotoxin concentration, heterotrophic plate count, and fecal coliform count.

Science.gov (United States)

Reyes, Mayra I; Pérez, Cynthia M; Negrón, Edna L

2008-03-01

Consumers increasingly use bottled water and home water treatment systems to avoid direct tap water. According to the International Bottled Water Association (IBWA), an industry trade group, 5 billion gallons of bottled water were consumed by North Americans in 2001. The principal aim of this study was to assess the microbial quality of in-house and imported bottled water for human consumption, by measurement and comparison of the concentration of bacterial endotoxin and standard cultivable methods of indicator microorganisms, specifically, heterotrophic and fecal coliform plate counts. A total of 21 brands of commercial bottled water, consisting of 10 imported and 11 in-house brands, selected at random from 96 brands that are consumed in Puerto Rico, were tested at three different time intervals. The Standard Limulus Amebocyte Lysate test, gel clot method, was used to measure the endotoxin concentrations. The minimum endotoxin concentration in 63 water samples was less than 0.0625 EU/mL, while the maximum was 32 EU/mL. The minimum bacterial count showed no growth, while the maximum was 7,500 CFU/mL. Bacterial isolates like P. fluorescens, Corynebacterium sp. J-K, S. paucimobilis, P. versicularis, A. baumannii, P. chlororaphis, F. indologenes, A. faecalis and P. cepacia were identified. Repeated measures analysis of variance demonstrated that endotoxin concentration did not change over time, while there was a statistically significant (p bacterial count over time. In addition, multiple linear regression analysis demonstrated that a unit change in the concentration of endotoxin across time was associated with a significant (p bacterial growth was not detected in some water samples, endotoxin was present. Measurement of Gram-negative bacterial endotoxins is one of the methods that have been suggested as a rapid way of determining bacteriological water quality.

Detecting swift fox: Smoked-plate scent stations versus spotlighting

Science.gov (United States)

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...

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A Molecular MST Approach to Investigate Fecal Indicator Bacteria in Bioaerosols, Bathing Water, Seaweed Wrack, and Sand at Recreational Beaches

Science.gov (United States)

Thoren, K. M.; Sinigalliano, C. D.

2016-02-01

Despite numerous cases of beach bacteria affecting millions of people worldwide, the persistence of the bacteria populations in coastal areas is still not well understood. The purpose of this study was to test the levels of persistence of Fecal Indicating Bacteria (FIB) of enterococci, Escherichia coli, and Human-source Bacteroidales, within the intertidal "swash zone" and the deeper waist zone in which people commonly bathe and play. In addition, the study sought to determine if these bacterial contaminants may also be found in aerosols at the beach. Measuring solar insolation in relation to bacterial persistence in seaweed wrack was used to determine if sunlight plays a role in modifying concentrations of FIB at the beach. Light intensity measured by a solar photometer and air quality measured by aerosol plate counts and qPCR Microbial Source Tracking (MST) was compared to varying locations where the beach samples were collected. Results from water samples demonstrate that bacteria measured using plate counts and qPCR were indeed higher within the swash zone than in the waist zone. This is in contrast with the way that the EPA currently measures and determines the public safety of beach waters. They commonly measure the waist zone, but disregard the swash zone. Results from beach bio-aerosol samples showed a wide variety of fungi and bacteria in the beach air, and qPCR MST analysis of these bio-aerosols showed the presence of FIBs such as enterococci on several of the aerosol collection plates. This emphasizes the need to collect samples from the entire beach instead of just measuring at an isolated area, and that exposure to microbial contaminants may include bathing water, beach sand, seaweed wrack, and bio-aerosols. Thus, the data reveals a potential way to identify harmful levels of bacteria and dangerous levels of poor air quality at recreational beaches. These results expound the need for broader assessment of potential beach contamination, not only the

Prevalence of pathogenic bacteria in street vended ready-to-eat meats in Windhoek, Namibia.

Science.gov (United States)

Shiningeni, Daphney; Chimwamurombe, Percy; Shilangale, Renatus; Misihairabgwi, Jane

2018-05-31

To determine the prevalence of pathogenic bacteria in street vended ready-to-eat meats in Windhoek, Namibia, a total of 96 street vended ready to eat meat samples were evaluated. Prevalences of 42%, 52%, 15%, 6% and 83% were observed for Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Shigella and Enterobacteriaceae respectively, while the highest aerobic plate counts were 7.74 Log 10 cfu/g, 5.67 Log 10 cfu/g, 5.12 Log 10 cfu/g , 4.56 Log 10 cfu/g, 3.3 Log 10 cfu/g, 5.75 Log 10 cfu/g respectively. Unsatisfactory microbial levels were 32% for aerobic plate count, 26% for Enterobacteriaceae, 35% for Escherichia coli, 11% for Listeria monocytogenes, 7% for Staphylococcus aureus and 6% for Shigella. Salmonella was detected in 11% and 40% of samples from two suburbs. The unsatisfactory microbiological quality of some ready-to-eat meats necessitates the provision of training on food safety and hygiene to street vendors for consumer protection. Copyright © 2018 Elsevier Ltd. All rights reserved.

High Reproducibility of ELISPOT Counts from Nine Different Laboratories

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Srividya Sundararaman

2015-01-01

Full Text Available The primary goal of immune monitoring with ELISPOT is to measure the number of T cells, specific for any antigen, accurately and reproducibly between different laboratories. In ELISPOT assays, antigen-specific T cells secrete cytokines, forming spots of different sizes on a membrane with variable background intensities. Due to the subjective nature of judging maximal and minimal spot sizes, different investigators come up with different numbers. This study aims to determine whether statistics-based, automated size-gating can harmonize the number of spot counts calculated between different laboratories. We plated PBMC at four different concentrations, 24 replicates each, in an IFN-γ ELISPOT assay with HCMV pp65 antigen. The ELISPOT plate, and an image file of the plate was counted in nine different laboratories using ImmunoSpot® Analyzers by (A Basic Count™ relying on subjective counting parameters set by the respective investigators and (B SmartCount™, an automated counting protocol by the ImmunoSpot® Software that uses statistics-based spot size auto-gating with spot intensity auto-thresholding. The average coefficient of variation (CV for the mean values between independent laboratories was 26.7% when counting with Basic Count™, and 6.7% when counting with SmartCount™. Our data indicates that SmartCount™ allows harmonization of counting ELISPOT results between different laboratories and investigators.

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

Science.gov (United States)

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 pulse stacking method of particle counting applied to position sensitive detection

International Nuclear Information System (INIS)

Basilier, E.

1976-03-01

A position sensitive particle counting system is described. A cyclic readout imaging device serves as an intermediate information buffer. Pulses are allowed to stack in the imager at very high counting rates. Imager noise is completely discriminated to provide very wide dynamic range. The system has been applied to a detector using cascaded microchannel plates. Pulse height spread produced by the plates causes some loss of information. The loss is comparable to the input loss of the plates. The improvement in maximum counting rate is several hundred times over previous systems that do not permit pulse stacking. (Auth.)

Truce with oxygen - A naerobiosis outcompete aerobiosis in the Antarctic lacustrine bacteria

Digital Repository Service at National Institute of Oceanography (India)

LokaBharathi, P.A.; Nair, S.; DeSouza, M.J.B.D.; Chandramohan, D.

. All colonies that were not black by sulfide precipitation were counted as FB. The anaerobic CFU were AnB, FB, SRB, and TDLO. The plates were incubated at 8–10° C for 10–20 days and the tubes for ca 30 days. All samples have been analysed... aerobiosis in the Antarctic lacustrine bacteria Page 3 of 5 file://C:\\My Documents\\articles23.htm 2/11/05 sulfide (63–188 ppm) at 5° C for 16 h and the viability estimated using Kogure’s method10,11. Viability is expressed as percentage of total viable...

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Full Text Available ... 800-342-2383) Give by Mail Close ... your plate with more non-starchy veggies and smaller portions of starchy foods and protein—no special tools or counting required! You can ...

Isolation and Identification of Phosphate Solubilizing and Nitrogen Fixing Bacteria from Soil in Wamena Biological Garden, Jayawijaya, Papua

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SRI WIDAWATI

2005-07-01

Full Text Available A study was undertaken to investigate the occurrence of phosphate solubilizing bacteria (PSB and nitrogen-fixing bacteria (NFB from soil samples of Wamena Biological Garden (WbiG. Eleven soil samples were collected randomly to estimate microbial population which used plate count method. The result showed that the microbial population ranged from 5.0x103-7.5x106 cells of bacteria/gram of soil and 5.0x103-1.5x107 cells of bacteria/gram of soil for PSB and NFB respectively. There were 17 isolates which have been identified till genus and species. The isolated microorganism were identified as PSB i.e. Bacillus sp., B. pantothenticus, B. megatherium, Flavobacterium sp., F. breve, Klebsiella sp., K. aerogenes, Chromobacterium lividum, Enterobacter alvei, E. agglomerans, Pseudomonas sp., Proteus sp. and as NFB i.e. Azotobacter sp., A. chroococcum, A. paspalii, Rhizobium sp., and Azospirillum sp.

Isolation and Characterization of Pb Resistant Bacteria from Cilalay Lake, Indonesia

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Kesi Kurnia

2015-12-01

Full Text Available Pollution of water environment with heavy metals is becoming one of the most severe environmental and human health hazards. Lead (Pb is a major pollutant and highly toxic to human, animals, plants, and microbes. Toxic metals are difficult to remove from the environment, since they cannot be chemically or biologically degraded and are ultimately indestructible. Biological approaches based on metal-resistant microorganisms have received a great deal of attention as alternative remediation processes. This study aim to isolate and characterize Pb resistant of heterotrophic bacteria in Cilalay Lake, West Java, Indonesia. The water samples were collected along three points around Cilalay Lake. Water physical and chemical determination was performed using the Water Quality Checker. The bacterial isolates were screened on Triptone Glucose Yeast (TGY agar plates. Afterwards selected isolates were grown on Nutrient Agar media 50% with supplemented Pb 100 ppm by the standard disk. Population of resistant bacteria was counted. The result from metal resistant bacteria indicated that all isolates were resistant. The most abundant type of resistant bacteria to lead was Gram negative more than Gram positive. Identified have metal resistant bacteria could be useful for the bioremediation of heavy metal contaminated sewage and waste water

Single ion counting with a MCP (microchannel plate) detector

Energy Technology Data Exchange (ETDEWEB)

Tawara, Hiroko; Sasaki, Shinichi; Miyajima, Mitsuhiro [National Lab. for High Energy Physics, Tsukuba, Ibaraki (Japan); Shibamura, Eido

1996-07-01

In this study, a single-ion-counting method using alpha-particle-impact ionization of Ar atoms is demonstrated and the preliminary {epsilon}{sub mcp} for Ar ions with incident energies of 3 to 4.7 keV is determined. The single-ion counting by the MCP is aimed to be performed under experimental conditions as follows: (1) A signal from the MCP is reasonably identified as incidence of single Ar-ion. (2) The counting rate of Ar ions is less than 1 s{sup -1}. (3) The incident Ar ions are not focused on a small part of an active area of the MCP, namely, {epsilon}{sub mcp} is determined with respect to the whole active area of the MCP. So far, any absolute detection efficiency has not been reported under these conditions. (J.P.N.)

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

Science.gov (United States)

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.

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Assessment of pathogenic bacteria in water and sediment from a water reservoir under tropical conditions (Lake Ma Vallée), Kinshasa Democratic Republic of Congo.

Science.gov (United States)

Mwanamoki, Paola M; Devarajan, Naresh; Thevenon, Florian; Atibu, Emmanuel K; Tshibanda, Joseph B; Ngelinkoto, Patience; Mpiana, Pius T; Prabakar, Kandasamy; Mubedi, Josué I; Kabele, Christophe G; Wildi, Walter; Poté, John

2014-10-01

This study was conducted to assess potential human health risks presented by pathogenic bacteria in a protected multi-use lake-reservoir (Lake Ma Vallée) located in west of Kinshasa, Democratic Republic of Congo (DRC). Water and surface sediments from several points of the Lake were collected during summer. Microbial analysis was performed for Escherichia coli, Enterococcus (ENT), Pseudomonas species and heterotrophic plate counts. PCR amplification was performed for the confirmation of E. coli, ENT, Pseudomonas spp. and Pseudomonas aeruginosa isolated from samples. The results reveal low concentration of bacteria in water column of the lake, the bacterial quantification results observed in this study for the water column were below the recommended limits, according to WHO and the European Directive 2006/7/CE, for bathing water. However, high concentration of bacteria was observed in the sediment samples; the values of 2.65 × 10(3), 6.35 × 10(3), 3.27 × 10(3) and 3.60 × 10(8) CFU g(-1) of dry sediment for E. coli, ENT, Pseudomonas spp. and heterotrophic plate counts, respectively. The results of this study indicate that sediments of the Lake Ma Vallée can constitute a reservoir of pathogenic microorganisms which can persist in the lake. Possible resuspension of faecal indicator bacteria and pathogens would affect water quality and may increase health risks to the population during recreational activities. Our results indicate that the microbial sediment analysis provides complementary and important information for assessing sanitary quality of surface water under tropical conditions.

Removal of crude petroleum hydrocarbons by heterotrophic bacteria ...

African Journals Online (AJOL)

Nitrogenous fertilizer (NPK) plant effluents from NAFCON were used in amending plots of land experimentally polluted with crude oil. Counts of heterotrophic bacteria (THBC) and fungi (TF), and of petroleum utilizing bacteria (PUB) and fungi (PUF) were monitored during an 8 weeks period. Counts obtained showed that ...

Population of bacteria from soil in Tudu-Aog village, Passi district, Bolaang Mongondow, North Sulawesi

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RIANI HARDININGSIH

2004-01-01

Full Text Available An experiment was conducted in order to know the population of bacteria from soil in Tudu-Aog village, Passi district, Bolaang Mongondow, North Sulawesi, the purpose of the research was to study the population of bacteria from soil. Fourthy six soil samples were taken from two location, namelyTudu-Aog village and Bugis mountain. Isolation was done by dilution methods on YEMA medium (for Rhizobium bacteria, Winogradsky’s (for Azotobacter bacteria, Pycosvkaya (for Phosphat Solubilizing Bacteria, and selective Difco Pseudomonas (for Pseudomonas bacteria. Incubation at room temperature (27-280C until 15 days, and the enumeration with plate count method. The highest enumeration of Rhizobium bacteria with plant rhizosphere of Alocasia esculenta (27x105 CFU/g soil, Theobroma cacao (29x105 CFU/g soil,and Euphorbia paniculata (26x105 CFU/g soil, Azotobacter bacteria with plant rhizosphere of Lycopersicum esculantum (38x105 CFU/g soil, Eugenia aromaticum (43x105 CFU/g soil, Andropogon sp. (34x105 CFU/g soil, Phosphat Solubilizing bacteria with plant rhizosphere of Sechium edule (27x105 CFU/g soil, Cinnamomum sp. (48x105 CFU/g soil, Cyathea sp. (72x105 CFU/g soil, and Pseudomonas bacteria with plant rhizosphere of Oryza sativa (18x105 CFU/g soil, Vanilla sp. (12x105 CFU/g soil, dan Saurauia sp.(19x105 CFU/g soil.

Measurement of environmental radioactivity with photo-diode and Imaging Plate

International Nuclear Information System (INIS)

Mori, C.; Sumi, T.; Gotoh, S.; Saze, T.; Nishizawa, K.

2000-01-01

Measurement of environmental radioactivity with photo-diode (PD) and Imaging Plate (IP) was tired. Commercially available Si PIN type PD's generally have depletion layer thickness more than a few hundred micrometer, which is enough for alpha particle measurement. PD's have various features: being usable in normal temperature, high energy resolution and low cost. Radon daughter nuclides positively charged in atmosphere were collected on the PD surface with negative electric potential and measured the pulse height spectra of alpha-particles from the daughter nuclides of Radon in thorium oxide, uranium ore, granite, and concrete. Counting of alpha-particles with IP was tired. Lead plates usually contain Pb-210 (RaD) and emit alpha-particles from Po-210. The alpha-particles from the plate were counted with PD and the plate was exposed to IP. By adjusting the gradation level on the reading out of the latent image, it was possible to count alpha-particle incident image one by one, and the number per 1 cm 2 was compared with the number of count with PD. (author)

Using flow cytometry for counting natural planktonic bacteria and understanding the structure of planktonic bacterial communities

Directory of Open Access Journals (Sweden)

Josep M. Gasol

2000-06-01

Full Text Available Flow cytometry is rapidly becoming a routine methodology in aquatic microbial ecology. The combination of simple to use bench-top flow cytometers and highly fluorescent nucleic acid stains allows fast and easy determination of microbe abundance in the plankton of lakes and oceans. The different dyes and protocols used to stain and count planktonic bacteria as well as the equipment in use are reviewed, with special attention to some of the problems encountered in daily routine practice such as fixation, staining and absolute counting. One of the main advantages of flow cytometry over epifluorescence microscopy is the ability to obtain cell-specific measurements in large numbers of cells with limited effort. We discuss how this characteristic has been used for differentiating photosynthetic from non-photosynthetic prokaryotes, for measuring bacterial cell size and nucleic acid content, and for estimating the relative activity and physiological state of each cell. We also describe how some of the flow cytometrically obtained data can be used to characterize the role of microbes on carbon cycling in the aquatic environment and we prospect the likely avenues of progress in the study of planktonic prokaryotes through the use of flow cytometry.

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

Science.gov (United States)

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.

DETERMINATION OF MINERAL CONTAIN AND BACTERIA CONTAMINANT ON ORGANIC AND NONORGANIC FRESH VEGETABLES

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Harsojo Harsojo

2010-06-01

Full Text Available The determination of mineral content and bacteria contaminant on fresh vegetable of long bean (Vegan ungulate Wall., white cabbage (Basic tolerance L., and lettuce (Lectuca sativa L. that cultivated by organic and nonorganic system have been done. The mineral content has been analyzed using neutron activation analysis and atomic absorption spectroscopy method, while bacteria contaminant by total plate count number using Nutrient Agar, Mac Conkey Agar, Baird Parker medium, and Salmonella using selective medium. The results showed that there are some essential mineral such as Fe, Zn, Ca, Co, and nonessential mineral Cd. There is tendency that fresh vegetable that cultivated by organic system contained Fe, Zn, Ca, Co and Cd mineral less than nonorganic. The Zn mineral content in nonorganic of fresh vegetable were higher than the limit of threshold number from Health Department, Republic of Indonesia (2004, while Cd mineral in organic or nonorganic of fresh vegetable were greater then threshold number from Codex Alimentarius Commision. The measurement of bacteria contaminant on organic and nonorganic of fresh vegetables contained aerob, coli, and Staphylococcus bacteria in organic of fresh vegetables were less compared to nonorganic of fresh vegetables.   Keywords: mineral, bacteria aerob, coli, Staphylococcus, Salmonella, organic, and nonorganic vegetable, neutron activation

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.

Science.gov (United States)

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

Effects of combined sewer overflow and stormwater on indicator bacteria concentrations in the Tama River due to the high population density of Tokyo Metropolitan area.

Science.gov (United States)

Ham, Young-Sik; Kobori, Hiromi; Takasago, Masahisa

2009-05-01

The indicator bacteria (standard plate count, total coliform, and fecal coliform bacteria) concentrations have been investigated using six ambient habitats (population density, percent sewer penetration, stream flow rate (m(3)/sec), percent residential area, percent forest area and percent agricultural area) in the Tama River basin in Tokyo, Japan during June 2003 to January 2005. The downstream and tributary Tama River showed higher concentrations of TC and FC bacteria than the upstream waters, which exceeded an environmental quality standard for rivers and a bathing water quality criterion. It was estimated that combined sewer overflow (CSO) and stormwater effluents contributed -4-23% to the indicator bacteria concentrations of the Tama River. The results of multiple regression analyses show that the indicator bacteria concentrations of Tama River basin are significantly affected by population density. It is concluded that the Tama River received a significant bacterial contamination load originating from the anthropogenic source.

Customized Computer Vision and Sensor System for Colony Recognition and Live Bacteria Counting in Agriculture

Directory of Open Access Journals (Sweden)

Gabriel M. ALVES

2016-06-01

Full Text Available This paper presents an arrangement based on a dedicated computer and charge-coupled device (CCD sensor system to intelligently allow the counting and recognition of colony formation. Microbes in agricultural environments are important catalysts of global carbon and nitrogen cycles, including the production and consumption of greenhouse gases in soil. Some microbes produce greenhouse gases such as carbon dioxide and nitrous oxide while decomposing organic matter in soil. Others consume methane from the atmosphere, helping to mitigate climate change. The magnitude of each of these processes is influenced by human activities and impacts the warming potential of Earth’s atmosphere. In this context, bacterial colony counting is important and requires sophisticated analysis methods. The method implemented in this study uses digital image processing techniques, including the Hough Transform for circular objects. The visual environment Borland Builder C++ was used for development, and a model for decision making was incorporated to aggregate intelligence. For calibration of the method a prepared illuminated chamber was used to enable analyses of the bacteria Escherichia coli, and Acidithiobacillus ferrooxidans. For validation, a set of comparisons were established between this smart method and the expert analyses. The results show the potential of this method for laboratory applications that involve the quantification and pattern recognition of bacterial colonies in solid culture environments.

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

Science.gov (United States)

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.

Disinfection of Escherichia coli bacteria using hybrid method of ozonation and hydrodynamic cavitation with orifice plate

Science.gov (United States)

Karamah, Eva F.; Ghaudenson, Rioneli; Amalia, Fitri; Bismo, Setijo

2017-11-01

This research aims to evaluate the performance of hybrid method of ozonation and hydrodynamic cavitation with orifice plate on E.coli bacteria disinfection. In this research, ozone dose, circulation flowrate, and disinfection method were varied. Ozone was produced by commercial ozonator with ozone dose of 64.83 mg/hour, 108.18 mg/hour, and 135.04 mg/hour. Meanwhile, hydrodynamic cavitation was generated by an orifice plate. The disinfection method compared in this research were: hydrodynamic cavitation, ozonation, and the combination of both. The best result on each method was achieved on the 60th minutes and with a circulation flowrate of 7 L/min. The hybrid method attained final concentration of 0 CFU/mL from the initial concentration of 2.10 × 105 CFU/mL. The ozonation method attained final concentration of 0 CFU/mL from the initial concentration of 1.32 × 105 CFU/mL. Cavitation method gives the least disinfection with final concentration of 5.20 × 104 CFU/mL from the initial concentration of 2.17 × 105 CFU/mL. In conclusion, hybrid method gives a faster and better disinfection of E.coli than each method on its own.

Utilization of bacteriocin-producing bacteria in dairy products

Directory of Open Access Journals (Sweden)

Matěj Patrovský

2016-07-01

Full Text Available Lactic acid bacteria have been used since ancient times for food preparation and for bio-conservation by fermentation. Selected strains are capable of producing antimicrobial peptides - bacteriocins, which can be natural preservatives, especially in products with short shelf lives. The present study is focused on inhibitory effects of the bacteriocin-producing bacteria strains Enterococcus faecium, Pediococccus acidilactici and Lactobacillus plantarum against Listeria innocua as an indicator microorganism. Freeze-dried preparations of bacterial strains producing particular bacteriocins were tested by agar well-diffusion assay and by the traditional spread plate method. Plantaricin exhibited the highest anti-listerial effect among the tested bacteriocins. Pediocin also demonstrated a distinct inhibitory effect, but enterocin appeared to be heat labile and its efficiency was also suppressed under cold storage conditions. Plantaricin reduced Listeria innocua counts by 1 log in dairy spread made from cheese and quark. The formation of bacteriocins by various Lactobacillus plantarum strains were substantially influenced by the cultivation conditions of the mother culture and by the microbial preparation process before freeze-drying. Bacteriocins introduced into foodstuffs via protective cultures in situ offer new perspectives on enhancing food quality and safety.

Parallel susceptibility testing of bacteria through culture-quantitative PCR in 96-well plates

Directory of Open Access Journals (Sweden)

Jun Luo

2018-05-01

Full Text Available Objective: The methods combining culture and quantitative PCR(qPCR offer new solutions for rapid antibiotic susceptibility testing(AST. However, the multiple steps of DNA extraction and cold storage of PCR reagents needed make them unsuitable for rapid high throughput AST. In this study, a parallel culture-qPCR method was developed to overcome above problems. Method: In this method, bacteria culture and DNA extraction automatically and simultaneously completed through using a common PCR instrument as a controllable heating device. A lyophilized 16S rDNA targeted qPCR reagent was also developed, which was stable and could be kept at 4 °C for long time and at 37 °C for about two months. Result: Testing of 36 P. aeruginosa isolates and 28 S. aureus isolates showed that the method had good agreements with the standard broth microdilution method, with an overall agreement of 97.22% (95% CI, 85.83–99.51 for P. aeruginosa and 96.43% (95% CI, 79.76–99.81 for S. aureus. This method could test 12 samples against a panel of up to 7 antibiotics simultaneously in two 96-well PCR plates within 4 h, which greatly improves the testing efficiency of the culture-qPCR method. Conclusion: With rapidness to obtain results and the capabilities for automation and multiple-sample testing, the parallel culture-qPCR method would have great potentials in clinical labs. Keywords: Antibiotic susceptibility testing, Thermo-cold lysis, Lyophilized qPCR reagent, Quantitative PCR, Bacteria

Isolation and Presumptive Identification of Adherent Epithelial Bacteria (“Epimural” Bacteria) from the Ovine Rumen Wall

OpenAIRE

Mead, Lorna J.; Jones, G. A.

1981-01-01

One hundred sixty-one strains of adherent bacteria were isolated under anaerobic conditions from four sites on the rumen epithelial surface of sheep fed hay or a hay-grain ration. Before isolation of bacteria, rumen tissue was washed six times in an anaerobic dilution solution, and viable bacteria suspended in the washings were counted. Calculation indicated that unattached bacteria would have been removed from the tissue by this procedure, but a slow and progressive release of attached bacte...

Microbial status of smoked fish, scombia scombia sold in Owerri, Imo ...

African Journals Online (AJOL)

These were analyzed microbiologically for viable heterotrophic bacteria and fungi count on Nutrient and Potato dextrose agar respectively, using pour plate method and coliform count in MacConkey broth by multiple tube method (MPN). The mean value ... Key words: Bacteria, Yeast, Mould, Smoked fish, Contamination.

Identification and discrimination of Pseudomonas aeruginosa bacteria grown in blood and bile by laser-induced breakdown spectroscopy

International Nuclear Information System (INIS)

Rehse, Steven J.; Diedrich, Jonathan; Palchaudhuri, Sunil

2007-01-01

Pseudomonas aeruginosa bacteria colonies have been analyzed by laser-induced breakdown spectroscopy using nanosecond laser pulses. LIBS spectra were obtained after transferring the bacteria from a nutrient-rich culture medium to a nutrient-free agar plate for laser ablation. To study the dependence of the LIBS spectrum on growth and environmental conditions, colonies were cultured on three different nutrient media: a trypticase soy agar (TSA) plate, a blood agar plate, and a medium chosen deliberately to induce bacteria membrane changes, a MacConkey agar plate containing bile salts. Nineteen atomic and ionic emission lines in the LIBS spectrum, which was dominated by inorganic elements such as calcium, magnesium and sodium, were used to identify and classify the bacteria. A discriminant function analysis was used to discriminate between the P. aeruginosa bacteria and two strains of E. coli: a non-pathogenic environmental strain and the pathogenic strain enterohemorrhagic E. coli 0157:H7 (EHEC). Nearly identical spectra were obtained from P. aeruginosa grown on the TSA plate and the blood agar plate, while the bacteria grown on the MacConkey plate exhibited easily distinguishable differences from the other two. All P. aeruginosa samples, independent of initial growth conditions, were readily discriminated from the two E. coli strains

Quicklime treatment and stirring of different poultry litter substrates for reducing pathogenic bacteria counts.

Science.gov (United States)

Lopes, M; Roll, V F B; Leite, F L; Dai Prá, M A; Xavier, E G; Heres, T; Valente, B S

2013-03-01

Testing different management practices can help to identify conditions that decrease or even eliminate pathogenic bacteria in poultry litter. A trial was conducted to evaluate the effects of daily manual stirring (rotation of the litter with a pitchfork) for the first 14 d of a bird's life (WDR), in 3 types of poultry litter substrates and quicklime treatment (CaO) during layout time between flocks on pathogenic bacteria occurrence (cfu). A total of 216 male Cobb broilers were randomly allotted to 18 pens with new litter (experimental unit). A split-plot design, with 6 treatments allotted to the main plots, was used: 1) wood shavings (WS) + WDR, 2) WS without stirring up to 14 d (WODR), 3) rice hulls (RIH) + WDR, 4) RIH + WODR, 5) mixture of 50% RIH and WS + WDR, and 6) mixture of 50% RIH and WS + WODR. Two treatments were allotted to the subplots: 0 and 300 g of CaO•m(-2) litter. After depopulation, litter samples were collected, and CaO was incorporated into the litter in the designated half of each pen. The cfu from litter samples after 7 d of the quicklime treatment were counted on Chapman agar, brain heart infusion media, and MacConkey agar. The data were analyzed using ANOVA, and the means were compared by least squares means (P litter efficiently reduced the cfu observed on brain heart infusion, Chapman agar, and MacConkey agar media by 57.2, 66.9, and 92.1%, respectively, compared with control (6.4, 17.9, and 46.1%; P litter reduces the cfu, regardless of the substrate and stirring performed.

Survival and Recovery of Methanotrophic Bacteria Starved Under Oxic and Anoxic Conditions

Science.gov (United States)

Roslev, Peter; King, Gary M.

1994-01-01

The effects of carbon deprivation on survival of methanotrophic bacteria were compared in cultures incubated in the presence and absence of oxygen in the starvation medium. Survival and recovery of the examined methanotrophs were generally highest for cultures starved under anoxic conditions as indicated by poststarvation measurements of methane oxidation, tetrazolium salt reduction, plate counts, and protein synthesis. Methylosinus trichosporium OB3b survived up to 6 weeks of carbon deprivation under anoxic conditions while maintaining a physiological state that allowed relatively rapid (hours) methane oxidation after substrate addition. A small fraction of cells starved under oxic and anoxic conditions (4 and 10%, respectively) survived more than 10 weeks but required several days for recovery on plates and in liquid medium. A non-spore-forming methanotroph, strain WP 12, displayed 36 to 118% of its initial methane oxidation capacity after 5 days of carbon deprivation. Oxidation rates varied with growth history prior to the experiments as well as with starvation conditions. Strain WP 12 starved under anoxic conditions showed up to 90% higher methane oxidation activity and 46% higher protein production after starvation than did cultures starved under oxic conditions. Only minor changes in biomass and niorpholow were seen for methanotrophic bacteria starved tinder anoxic conditions. In contrast, starvation under oxic conditions resulted in morphology changes and an initial 28 to 35% loss of cell protein. These data suggest that methanotrophic bacteria can survin,e carbon deprivation under anoxic conditions by using maintenance energy derived Solelyr from an anaerobic endogenous metabolism. This capability could partly explain a significant potential for methane oxidation in environments not continuously, supporting aerobic methanotrophic growth.

Rationalizing and advancing the 3-MPBA SERS sandwich assay for rapid detection of bacteria in environmental and food matrices.

Science.gov (United States)

Pearson, Brooke; Mills, Alexander; Tucker, Madeline; Gao, Siyue; McLandsborough, Lynne; He, Lili

2018-06-01

Bacterial foodborne illness continues to be a pressing issue in our food supply. Rapid detection methods are needed for perishable foods due to their short shelf lives and significant contribution to foodborne illness. Previously, a sensitive and reliable surface-enhanced Raman spectroscopy (SERS) sandwich assay based on 3-mercaptophenylboronic acid (3-MBPA) as a capturer and indicator molecule was developed for rapid bacteria detection. In this study, we explored the advantages and constraints of this assay over the conventional aerobic plate count (APC) method and further developed methods for detection in real environmental and food matrices. The SERS sandwich assay was able to detect environmental bacteria in pond water and on spinach leaves at higher levels than the APC method. In addition, the SERS assay appeared to have higher sensitivity to quantify bacteria in the stationary phase. On the other hand, the APC method was more sensitive to cell viability. Finally, a method to detect bacteria in a challenging high-sugar juice matrix was developed to enhance bacteria capture. This study advanced the SERS technique for real applications in environment and food matrices. Copyright © 2017 Elsevier Ltd. All rights reserved.

Single bioreactor gastrointestinal tract simulator for study of survival of probiotic bacteria.

Science.gov (United States)

Sumeri, Ingrid; Arike, Liisa; Adamberg, Kaarel; Paalme, Toomas

2008-08-01

The aim of the present study was to design an in vitro model system to evaluate the probiotic potential of food. A single bioreactor system-gastrointestinal tract simulator (GITS) was chosen for process simulation on account of its considerable simplicity compared to multi-vessel systems used in previous studies. The bioreactor was evaluated by studying the viability of four known probiotic bacteria (Lactobacillus acidophilus La-5, Lactobacillus johnsonii NCC 533, Lactobacillus casei strain Shirota, and Lactobacillus rhamnosus GG) as a function of their physiological state. L. acidophilus and L. johnsonii survived in GITS better when introduced at an early stationary or exponential phase compared to being previously stored for 2 weeks at 4 degrees C. These two species were more resistant to bile salts and survived better than L. casei and L. rhamnosus GG. The latter two species gave large losses (up to 6 log) in plate counts independent of growth state due to the bile. However, experiments with some commercial probiotic products containing Lb. GG bacteria showed much better survival compared with model food (modified deMan-Rogosa-Sharpe growth medium), thus demonstrating the influence of the food matrix on the viability of bacteria. The study demonstrated that GITS can be successfully used for evaluation of viability of probiotic bacteria and functionality of probiotic food.

Flow Cytometry Total Cell Counts: A Field Study Assessing Microbiological Water Quality and Growth in Unchlorinated Drinking Water Distribution Systems

Science.gov (United States)

Liu, G.; Van der Mark, E. J.; Verberk, J. Q. J. C.; Van Dijk, J. C.

2013-01-01

The objective of this study was to evaluate the application of flow cytometry total cell counts (TCCs) as a parameter to assess microbial growth in drinking water distribution systems and to determine the relationships between different parameters describing the biostability of treated water. A one-year sampling program was carried out in two distribution systems in The Netherlands. Results demonstrated that, in both systems, the biomass differences measured by ATP were not significant. TCC differences were also not significant in treatment plant 1, but decreased slightly in treatment plant 2. TCC values were found to be higher at temperatures above 15°C than at temperatures below 15°C. The correlation study of parameters describing biostability found no relationship among TCC, heterotrophic plate counts, and Aeromonas. Also no relationship was found between TCC and ATP. Some correlation was found between the subgroup of high nucleic acid content bacteria and ATP (R 2 = 0.63). Overall, the results demonstrated that TCC is a valuable parameter to assess the drinking water biological quality and regrowth; it can directly and sensitively quantify biomass, detect small changes, and can be used to determine the subgroup of active HNA bacteria that are related to ATP. PMID:23819117

Flow Cytometry Total Cell Counts: A Field Study Assessing Microbiological Water Quality and Growth in Unchlorinated Drinking Water Distribution Systems

Directory of Open Access Journals (Sweden)

G. Liu

2013-01-01

Full Text Available The objective of this study was to evaluate the application of flow cytometry total cell counts (TCCs as a parameter to assess microbial growth in drinking water distribution systems and to determine the relationships between different parameters describing the biostability of treated water. A one-year sampling program was carried out in two distribution systems in The Netherlands. Results demonstrated that, in both systems, the biomass differences measured by ATP were not significant. TCC differences were also not significant in treatment plant 1, but decreased slightly in treatment plant 2. TCC values were found to be higher at temperatures above 15°C than at temperatures below 15°C. The correlation study of parameters describing biostability found no relationship among TCC, heterotrophic plate counts, and Aeromonas. Also no relationship was found between TCC and ATP. Some correlation was found between the subgroup of high nucleic acid content bacteria and ATP (R2=0.63. Overall, the results demonstrated that TCC is a valuable parameter to assess the drinking water biological quality and regrowth; it can directly and sensitively quantify biomass, detect small changes, and can be used to determine the subgroup of active HNA bacteria that are related to ATP.

Bacteria associated with cultures of psathyrella atroumbonata (Pleger)

African Journals Online (AJOL)

These bacteria include Bacillus licheniformis, Bacillus subtilis, Leuconostoc mesenteroides, Pseudomonas aeruginosa, Bacillus cereus and Staphylococcus aureus. The average bacteria count was 1.0 x 106 cfu/ml and these bacteria grew within pH range of 5.0 and 9.0. the optimum temperature range of growth lied ...

Acoustic manipulation of bacteria cells suspensions

Science.gov (United States)

GutiéRrez-Ramos, Salomé; Hoyos, Mauricio; Aider, Jean Luc; Ruiz, Carlos; Acoustofluidics Team Team; Soft; Bio Group Collaboration

An acoustic contacless manipulation gives advantages in the exploration of the complex dynamics enviroment that active matter exhibits. Our works reports the control confinement and dispersion of Escherichia coliRP437-pZA3R-YFP suspensions (M9Glu-Ca) via acoustic levitation.The manipulation of the bacteria bath in a parallel plate resonator is achieved using the acoustic radiation force and the secondary radiation force. The primary radiation force generates levitation of the bacteria cells at the nodal plane of the ultrasonic standing wave generated inside the resonator. On the other side, secondary forces leads to the consolidation of stable aggregates. All the experiments were performed in the acoustic trap described, where we excite the emission plate with a continuous sinusoidal signal at a frequency in the order of MHz and a quartz slide as the reflector plate. In a typical experiment we observed that, before the input of the signal, the bacteria cells exhibit their typical run and tumble behavior and after the sound is turned on all of them displace towards the nodal plane, and instantaneously the aggregation begins in this region. CNRS French National Space Studies, CONACYT Mexico.

Eutectic cell and nodule count as the quality factors of cast iron

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E. Fraś

2008-10-01

Full Text Available In this work the predictions based on a theoretical analysis aimed at elucidating of eutectic cell count or nodule counts N wereexperimentally verified. The experimental work was focused on processing flake graphite and ductile iron under various inoculationconditions in order to achieve various physicochemical states of the experimental melts. In addition, plates of various wall thicknesses, s were cast and the resultant eutectic cell or nodule counts were established. Moreover, thermal analysis was used to find out the degree of maximum undercooling for the graphite eutectic, Tm. A relationship was found between the eutectic cell or nodule count and the maximum undercooling Tm.. In addition it was also found that N can be related to the wall thickness of plate shaped castings. Finally, the present work provides a rational for the effect of technological factors such as the melt chemistry, inoculation practice, and holding temperature and time on the resultant cell count or nodule count of cast iron. In particular, good agreement was found between the predictions of the theoretical analysis and the experimental data.

Undercooling, nodule count and carbides in thin walled ductile cast iron

DEFF Research Database (Denmark)

Pedersen, Karl Martin; Tiedje, Niels Skat

2008-01-01

Ductile cast iron has been cast in plate thicknesses between 2 to 8 mm. The temperature has been measured during the solidification and the graphite nodule count and size distribution together with the type and amount of carbides have been analysed afterwards. Low nodule count gives higher...

Survival of Antibiotic Resistant Bacteria and Horizontal Gene Transfer Control Antibiotic Resistance Gene Content in Anaerobic Digesters.

Science.gov (United States)

Miller, Jennifer H; Novak, John T; Knocke, William R; Pruden, Amy

2016-01-01

Understanding fate of antibiotic resistant bacteria (ARB) vs. their antibiotic resistance genes (ARGs) during wastewater sludge treatment is critical in order to reduce the spread of antibiotic resistance through process optimization. Here, we spiked high concentrations of tetracycline-resistant bacteria, isolated from mesophilic (Iso M1-1-a Pseudomonas sp.) and thermophilic (Iso T10-a Bacillus sp.) anaerobic digested sludge, into batch digesters and monitored their fate by plate counts and quantitative polymerase chain reaction (QPCR) of their corresponding tetracycline ARGs. In batch studies, spiked ARB plate counts returned to baseline (thermophilic) or 1-log above baseline (mesophilic) while levels of the ARG present in the spiked isolate [tet(G)] remained high in mesophilic batch reactors. To compare results under semi-continuous flow conditions with natural influent variation, tet(O), tet(W), and sul1 ARGs, along with the intI1 integrase gene, were monitored over a 9-month period in the raw feed sludge and effluent sludge of lab-scale thermophilic and mesophilic anaerobic digesters. sul1 and intI1 in mesophilic and thermophilic digesters correlated positively (Spearman rho = 0.457-0.829, P < 0.05) with the raw feed sludge. There was no correlation in tet(O) or tet(W) ratios in raw sludge and mesophilic digested sludge or thermophilic digested sludge (Spearman rho = 0.130-0.486, P = 0.075-0.612). However, in the thermophilic digester, the tet(O) and tet(W) ratios remained consistently low over the entire monitoring period. We conclude that the influent sludge microbial composition can influence the ARG content of a digester, apparently as a result of differential survival or death of ARBs or horizontal gene transfer of genes between raw sludge ARBs and the digester microbial community. Notably, mesophilic digestion was more susceptible to ARG intrusion than thermophilic digestion, which may be attributed to a higher rate of ARB survival and/or horizontal gene

Simultaneous aptasensor for multiplex pathogenic bacteria detection based on multicolor upconversion nanoparticles labels.

Science.gov (United States)

Wu, Shijia; Duan, Nuo; Shi, Zhao; Fang, Congcong; Wang, Zhouping

2014-03-18

A highly sensitive and specific multiplex method for the simultaneous detection of three pathogenic bacteria was fabricated using multicolor upconversion nanoparticles (UCNPs) as luminescence labels coupled with aptamers as the molecular recognition elements. Multicolor UCNPs were synthesized via doping with various rare-earth ions to obtain well-separated emission peaks. The aptamer sequences were selected using the systematic evolution of ligands by exponential enrichment (SELEX) strategy for Staphylococcus aureus, Vibrio parahemolyticus, and Salmonella typhimurium. When applied in this method, aptamers can be used for the specific recognition of the bacteria from complex mixtures, including those found in real food matrixes. Aptamers and multicolor UCNPs were employed to selectively capture and simultaneously quantify the three target bacteria on the basis of the independent peaks. Under optimal conditions, the correlation between the concentration of three bacteria and the luminescence signal was found to be linear from 50-10(6) cfu mL(-1). Improved by the magnetic separation and concentration effect of Fe3O4 magnetic nanoparticles, the limits of detection of the developed method were found to be 25, 10, and 15 cfu mL(-1) for S. aureus, V. parahemolyticus, and S. typhimurium, respectively. The capability of the bioassay in real food samples was also investigated, and the results were consistent with experimental results obtained from plate-counting methods. This proposed method for the detection of various pathogenic bacteria based on multicolor UCNPs has great potential in the application of food safety and multiplex nanosensors.

Low white blood cell count and cancer

Science.gov (United States)

... gov/ency/patientinstructions/000675.htm Low white blood cell count and cancer To use the sharing features on this page, please enable JavaScript. White blood cells (WBCs) fight infections from bacteria, viruses, fungi, and ...

Identification of antibiotic resistant bacteria community and a GeoChip based study of resistome in urban watersheds.

Science.gov (United States)

Low, Adrian; Ng, Charmaine; He, Jianzhong

2016-12-01

Urban watersheds from point sources are potential reservoirs of antibiotic resistance genes (ARGs). However, few studies have investigated urban watersheds of non-point sources. To understand the type of ARGs and bacteria that might carry such genes, we investigated two non-point source urban watersheds with different land-use profiles. Antibiotic resistance levels of two watersheds (R1, R3) were examined using heterotrophic plate counts (HPC) as a culturing method to obtain counts of bacteria resistant to seven antibiotics belonging to different classes (erythromycin, kanamycin, lincomycin, norfloxacin, sulfanilamide, tetracycline and trimethoprim). From the HPC study, 239 antibiotic resistant bacteria were characterized for resistance to more antibiotics. Furthermore, ARGs and antimicrobial biosynthesis genes were identified using GeoChip version 5.0 to elucidate the resistomes of surface waters in watersheds R1 and R3. The HPC study showed that water samples from R1 had significantly higher counts of bacteria resistant to erythromycin, kanamycin, norfloxacin, sulfanilamide, tetracycline and trimethoprim than those from R3 (Analysis of Similarity (ANOSIM), R = 0.557, p antibiotics tested, lincomycin and trimethoprim resistant bacteria are greater in abundances. The 239 antibiotic resistant isolates represent a subset of resistant bacterial populations, including bacteria not previously known for resistance. Majority of the isolates had resistance to ampicillin, vancomycin, lincomycin and trimethoprim. GeoChip revealed similar ARGs in both watersheds, but with significantly higher intensities for tetX and β-lactamase B genes in R1 than R3. The genes with the highest average normalized intensities in R1 and R3 were tetracycline (tet) and fosfomycin (fosA) resistance genes, respectively. The higher abundance of tetX genes in R1 is congruent with the higher abundance of tetracycline resistant HPC observed in R1 samples. Strong correlations (r ≥ 0.8) of efflux

Ammonia-oxidizing bacteria in a chloraminated distribution system: seasonal occurrence, distribution and disinfection resistance.

Science.gov (United States)

Wolfe, R L; Lieu, N I; Izaguirre, G; Means, E G

1990-02-01

Nitrification in chloraminated drinking water can have a number of adverse effects on water quality, including a loss of total chlorine and ammonia-N and an increase in the concentration of heterotrophic plate count bacteria and nitrite. To understand how nitrification develops, a study was conducted to examine the factors that influence the occurrence of ammonia-oxidizing bacteria (AOB) in a chloraminated distribution system. Samples were collected over an 18-month period from a raw-water source, a conventional treatment plant effluent, and two covered, finished-water reservoirs that previously experienced nitrification episodes. Sediment and biofilm samples were collected from the interior wall surfaces of two finished-water pipelines and one of the covered reservoirs. The AOB were enumerated by a most-probable-number technique, and isolates were isolated and identified. The resistance of naturally occurring AOB to chloramines and free chlorine was also examined. The results of the monitoring program indicated that the levels of AOB, identified as members of the genus Nitrosomonas, were seasonally dependent in both source and finished waters, with the highest levels observed in the warm summer months. The concentrations of AOB in the two reservoirs, both of which have floating covers made of synthetic rubber (Hypalon; E.I. du Pont de Nemours & Co., Inc., Wilmington, Del.), had most probable numbers that ranged from less than 0.2 to greater than 300/ml and correlated significantly with temperature and levels of heterotrophic plate count bacteria. No AOB were detected in the chloraminated reservoirs when the water temperature was below 16 to 18 degrees C. The study indicated that nitrifiers occur throughout the chloraminated distribution system. Higher concentrations of AOB were found in the reservoir and pipe sediment materials than in the pipe biofilm samples. The AOB were approximately 13 times more resistant to monochloramine than to free chlorine. After 33 min

hand- dug well

African Journals Online (AJOL)

Misrak

wells at the vicinity of three major refuse dumpsite taking measurement of depth of the well, diameter, volume of water, distance ... The total coliform bacteria count shows that the water is exposed to serious and complex contaminations, which .... Bacterial plate count was carried out using the pour plate method with nutrient ...

THE PHOTODYNAMIC EFFECT OF LED-MAGNETIC EXPOSURE TO PHOTOINACTIVATION OF AEROBIC PHOTOSYNTETIC BACTERIA

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Suryani Dyah Astuti

2014-01-01

Full Text Available All photosynthetic bacteria have a major pigment of bacteriochlorophyl and accessor pigment e.g. the carotenoids, which both have an important role in photosynthesis process. This study aim to explore the exogenous organic photosensitizer from photosyntetic bacteria for photodynamic therapy application. This study is an experimental research aiming to test the potential illumination ofLED with wavelength 409, 430, 528 and 629 nm, and power optimization and time exposure LED-magnetic for optimum photo activation Rhodococcus growth. The reseach design use a factorial completely randomized design with factor ofpower and exposure time. The number ofbacterial colonies grown measure using ofTotal Plate Count (TPC methods. The result ofanova test shows that irradiation treatment with LED 409 nm, 430 nm, 528 nm and 629 nm significantly affects on bacterial colony growth. LED 409 nm exposure has the greatest potential to boost the growth ofbacterial colonies by 77%. LED exposure and the addition of1.8 mT magnetic field increases bacterial colony growth by 98%. Results of optimization of LED and magnetic fields show power 46 mW and a 40 minute (energy dose 110 J/cm2 optimum growth ofbacterial colonies increase by 184%. So LED and magnetic illumination has potentially increased the viability ofan aerob photosyntetic bacteria colonies.

Damage mechanisms of pathogenic bacteria in drinking water ...

African Journals Online (AJOL)

This study aimed at elucidating the inactivation mechanisms of pathogenic bacteria in drinking water during chlorine and solar disinfection using a simple plating method. The well-known bacterial model Escherichia coli was used as pathogenic bacteria for the experiments. The damage mechanisms of E. coli were ...

Effect of Gaseous Ozone Exposure on the Bacteria Counts and Oxidative Properties of Ground Hanwoo Beef at Refrigeration Temperature.

Science.gov (United States)

Cho, Youngjae; Muhlisin; Choi, Ji Hye; Hahn, Tae-Wook; Lee, Sung Ki

2014-01-01

This study was designed to elucidate the effect of ozone exposure on the bacteria counts and oxidative properties of ground Hanwoo beef contaminated with Escherichia coli O157:H7 at refrigeration temperature. Ground beef was inoculated with 7 Log CFU/g of E. coli O157:H7 isolated from domestic pigs and was then subjected to ozone exposure (10×10(-6) kg O3 h(-1)) at 4℃ for 3 d. E. coli O157:H7, total aerobic and anaerobic bacterial growth and oxidative properties including instrumental color changes, TBARS, catalase (CAT) and glutathione peroxidase (GPx) activity were evaluated. Ozone exposure significantly prohibited (pgenerator inside a refrigerator. Further studies regarding the ozone concentrations and exposure times are needed.

Enhanced counting efficiency of Cerenkov radiation from bismuth-210

International Nuclear Information System (INIS)

Peck, G.A.; Smith, J.D.

1998-01-01

This paper describes the measurement of 210 Bi by Cerenkov counting in a commercial liquid scintillation counter. The counting efficiency in water is 0.17 counts per second per Becquerel (17%). When the enhancers Triton X-100 (15% v/v) and sodium salicylate (1% m/v) are added to the solution the counting efficiency for 210 Bi increases from 17% to 75%. The 210 Po daughter of 210 Bi causes interference of 0.85 counts per second per Becquerel in the presence of the enhancers but not in water. When 210 Bi and 210 Po are present in secular equilibrium the total counting efficiency is 160%. When 210 Bi and 210 Po are not in secular equilibrium the 210 Po can be removed immediately before counting by plating onto silver foil. The use of the enhancers gives a substantial increase in counting efficiency compared to counting in water. Compared with solutions used in liquid scintillation counting the enhancer solution is inexpensive and can be disposed of without environmental hazard. (author)

The increase in leucocyte count during menstruation is a function of ...

African Journals Online (AJOL)

Menstruation is an endocrine mediated physiologic cyclical bleeding per vagina in a non-pregnant woman of the reproductive age group. Blood is a good culture medium for bacteria and as such, menstruation can affect leucocytes count. This study is aimed at evaluating leucocytes count during menstruation. Fifty-eight ...

Total bacterial count and somatic cell count in refrigerated raw milk stored in communal tanks

Directory of Open Access Journals (Sweden)

Edmar da Costa Alves

2014-09-01

Full Text Available The current industry demand for dairy products with extended shelf life has resulted in new challenges for milk quality maintenance. The processing of milk with high bacterial counts compromises the quality and performance of industrial products. The study aimed to evaluate the total bacteria counts (TBC and somatic cell count (SCC in 768 samples of refrigerated raw milk, from 32 communal tanks. Samples were collected in the first quarter of 2010, 2011, 2012 and 2013 and analyzed by the Laboratory of Milk Quality - LQL. Results showed that 62.5%, 37.5%, 15.6% and 27.1% of the means for TBC in 2010, 2011, 2012 and 2013, respectively, were above the values established by legislation. However, we observed a significant reduction in the levels of total bacterial count (TBC in the studied periods. For somatic cell count, 100% of the means indicated values below 600.000 cells/mL, complying with the actual Brazilian legislation. The values found for the somatic cell count suggests the adoption of effective measures for the sanitary control of the herd. However, the results must be considered with caution as it highlights the need for quality improvements of the raw material until it achieves reliable results effectively.

Effect of the Intelligent Health Messenger Box on health care professionals' knowledge, attitudes, and practice related to hand hygiene and hand bacteria counts.

Science.gov (United States)

Saffari, Mohsen; Ghanizadeh, Ghader; Fattahipour, Rasoul; Khalaji, Kazem; Pakpour, Amir H; Koenig, Harold G

2016-12-01

We assessed the effectiveness of the Intelligent Health Messenger Box in promoting hand hygiene using a quasiexperimental design. Knowledge, attitudes, and self-reported practices related to hand hygiene as well as hand bacteria counts and amount of liquid soap used were measured. The intervention involved broadcasting preventive audio messages. All outcomes showed significant change after the intervention compared with before. The Intelligent Health Messenger Box can serve as a practical way to improve hand hygiene. Copyright © 2016 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

Method Verification Requirements for an Advanced Imaging System for Microbial Plate Count Enumeration.

Science.gov (United States)

Jones, David; Cundell, Tony

2018-01-01

The Growth Direct™ System that automates the incubation and reading of membrane filtration microbial counts on soybean-casein digest, Sabouraud dextrose, and R2A agar differs only from the traditional method in that micro-colonies on the membrane are counted using an advanced imaging system up to 50% earlier in the incubation. Based on the recommendations in USP Validation of New Microbiological Testing Methods , the system may be implemented in a microbiology laboratory after simple method verification and not a full method validation. LAY ABSTRACT: The Growth Direct™ System that automates the incubation and reading of microbial counts on membranes on solid agar differs only from the traditional method in that micro-colonies on the membrane are counted using an advanced imaging system up to 50% earlier in the incubation time. Based on the recommendations in USP Validation of New Microbiological Testing Methods , the system may be implemented in a microbiology laboratory after simple method verification and not a full method validation. © PDA, Inc. 2018.

Comparison of apical extrusion of intracanal bacteria by various glide-path establishing systems: an in vitro study

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Alberto Dagna

2017-11-01

Full Text Available Objectives This study compared the amount of apically extruded bacteria during the glide-path preparation by using multi-file and single-file glide-path establishing nickel-titanium (NiTi rotary systems. Materials and Methods Sixty mandibular first molar teeth were used to prepare the test apparatus. They were decoronated, blocked into glass vials, sterilized in ethylene oxide gas, infected with a pure culture of Enterococcus faecalis, randomly assigned to 5 experimental groups, and then prepared using manual stainless-steel files (group KF and glide-path establishing NiTi rotary files (group PF with PathFiles, group GF with G-Files, group PG with ProGlider, and group OG with One G. At the end of canal preparation, 0.01 mL NaCl solution was taken from the experimental vials. The suspension was plated on brain heart infusion agar and colonies of bacteria were counted, and the results were given as number of colony-forming units (CFU. Results The manual instrumentation technique tested in group KF extruded the highest number of bacteria compared to the other 4 groups (p < 0.05. The 4 groups using rotary glide-path establishing instruments extruded similar amounts of bacteria. Conclusions All glide-path establishment instrument systems tested caused a measurable apical extrusion of bacteria. The manual glide-path preparation showed the highest number of bacteria extruded compared to the other NiTi glide-path establishing instruments.

External Microflora of a Marine Wood-Boring Isopod

OpenAIRE

Boyle, Paul J.; Mitchell, Ralph

1981-01-01

Bacteria associated with the marine wood-boring isopod Limnoria lignorum were enumerated by acridine orange epifluorescence microscopy and by plate counts on several media; the plate-viable bacteria were isolated and identified. Similar procedures were followed to enumerate and identify bacteria associated with the wood substrate from which the isopods were collected and with the surrounding water from the isopod habitat. Approximately 1.4 × 107 bacterial cells were associated with each indiv...

Coffee Flower Honey (Coffea sp. Addition to Kefir Quality Based on Microbiology Characteristic

Directory of Open Access Journals (Sweden)

Astri Wulandari

2017-11-01

Full Text Available The Objective of this research was to determine the best concentration of addition coffee flower honey to kefir based on total plate count, lactic acid bacteria, acidity, and pH value. The materials used in this research were honey kefir made from cow’s milk, kefir grains, and coffee flowers honey. The method used in this research was experimental with Completely Randomized Design by 4 treatments and 4 replications and continued by using Duncan’s Multiple Range Test. The result showed that the effect of addition coffee flowers honey was not significantly different (P>0.05 on total plate count, lactic acid bacteria, acidity, give highly significant different (P<0.01 on pH value. Addition 20% of  coffee flower honey was the best treatment with quality of total plate count 6.21 ± 0.60 log CFU/mL, total lactic acid bacteria 6.16 ± 0.14 log CFU/mL, acidity 0.76 ± 0.11 % and pH value 3.596 ± 0.021. Further study is needed to figure the storability of honey kefir.

Unusual rise in mercury-resistant bacteria in coastal environs

Digital Repository Service at National Institute of Oceanography (India)

Ramaiah, N.; De, J.

A sharp rise in mercury-resistant bacteria (MRB) capable of tolerating very high concentration of Hg was observed over the last 3-4 years in the coastal environs of India. While none or negligible colony-forming units (CFU) of bacteria were counted...

Okara: A Nutritionally Valuable By-product Able to Stabilize Lactobacillus plantarum during Freeze-drying, Spray-drying, and Storage

Science.gov (United States)

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

Some physiological and morphological aspects of radiation-resistant bacteria and a new method for their isolation from food

International Nuclear Information System (INIS)

Sanders, S.W.

1978-01-01

A study was undertaken to help clarify the taxonomic positions and mechanisms of radiation resistance of radiation-resistant asporogenous bacteria found in foods. Determinations of DNA base compositions of highly resistant Moroxella-Acinetobacter (M-A) strains indicated that they were atypical, having percent guanine plus cytosine contents exceeding the values for true Moraxella or Acinetobacter spp. By direct observation of dividing cells, the resistant M-A were found to undergo multiple-plane division. Electron micrographs revealed unusually thick cell walls in the M-A as compared with gram-negative bacteria, indicating a possible role of the cell wall in radiation resistance. Resistance to desiccation was utilized in the selection of highly radiation-resistant bacteria from non-irradiated sources. Bacteria from a food or other source were suspended in dilute phosphate buffer and dried in a thin film at 25 C and 33% relative humidity. Storage under these conditions for 15 days or more reduced the numbers of radiation-sensitive bacteria. Further selection of the most radiation-resistant bacteria was obtained by irradiation of bacteria on velveteen in the replication process, thereby allowing the isolation of highly resistant bacteria that had not been irradiated. The similarity of radiation-resistance and identifying characteristics between irradiated and non-irradiated isolates indicated that highly radiation-resistant bacteria are not altered by radiation selection. Irradiated Plate Count Agar and Tryptic Soy Agar were found to be very toxic to radiation-resistant bacteria. This phenomenon may be important in food irradiation, where the ability to survive and grow in a product may depend partly on the sensitivity to bacteriocidal products formed during irradiation

Preparation of actinide targets by molecular plating for Coulomb excitation studies at ATLAS

International Nuclear Information System (INIS)

Greene, J. P.

1998-01-01

Molecular plating is now routinely used to prepare sources and targets of actinide elements. Although the technique is simple and fairly reproducible, because of the radioactive nature of the target it is very useful to record various parameters in the preparation of such targets. At Argonne, ∼200 microg/cm 2 thick targets of Pu and Cm were required for Coulomb Excitation (COULEX) Studies with the Argonne-Notre Dame BGO gamma ray facility and later with the GAMMASPHERE. These targets were plated on 50 mg/cm 2 Au backing and were covered with 150 microg/cm 2 Au foil. Targets of 239 Pu, 240 Pu, 242 Pu, 244 Pu and 248 Cm were prepared by dissolving the material in isopropyl alcohol and electroplating the actinide ions by applying 600 volts. The amount of these materials on the target was determined by alpha particle counting and gamma ray counting. Details of the molecular plating and counting will be discussed

An improved plating assay for determination of phage titer | Yang ...

African Journals Online (AJOL)

In this study, an improved plating assay was developed for detection of the number of recombinant phage Cap-T7 present in a test solution at a certain dilution point by counting the plaque forming units. The data demonstrated that the improved plating assay is fast, useful, and convenient for the determination of the phage ...

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

Science.gov (United States)

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.

Bacteriological Analysis and Hygine Level of Food Outlets within Rufus Giwa Polytechnic, Owo, Ondo State, Nigeria.

OpenAIRE

Ibrahim TA; Akenroye OM; Osabiya OJ

2013-01-01

The bacteriological quality of three major food outlets in Rufus Giwa Polytechnic, Owo, was assessed using standard bacteriological methods. Swabs of hands of food vendors, table and plates in these outlets were assessed for total bacterial count, total coliform count and total E. coli count. A total of 789 bacterial colonies were isolated from hands of food handlers, tables and plates used for eating in the outlets. Eleven genera of bacteria were isolated and identified, they were; klebsiell...

Study on auto-plating process time versus recovery for polonium, Po-210 in environmental sample

International Nuclear Information System (INIS)

Jalal Sharib; Zaharudin Ahmad; Abdul Kadir Ishak; Norfaizal Mohamed; Ahmad Sanadi Abu Bakar; Yii Mei Wo; Kamarozaman Ishak; Siti Aminah Yusoff

2008-08-01

This study was carried out to evaluate time effectiveness and recovery 16 samples of 4 Kuala Muda stations during auto-plating process procedures for determination Polonium, Po 210 activity concentration in environmental sample. The study was performed using Kuala Muda sediment as sample in the same methodology. The auto-plating process runs for 4, 12, 24 and 30 hours on a silver disc for 4 samples each station, and then counted for one (1) day using an alpha spectrometry counting system. The objectives for this study is to justify on time duration for auto-plating process effecting a chemical yield of Po-209.The results showed recovery are increasing versus time and constantly at 24 hour auto-plating. Its mean, 24 hour is an optimum time for auto-plating process for determination of Polonium, Po 210 activity concentration in environmental sample. (Author)

Rapid fluorescence detection of pathogenic bacteria using magnetic enrichment technique combined with magnetophoretic chromatography.

Science.gov (United States)

Che, Yulan; Xu, Yi; Wang, Renjie; Chen, Li

2017-08-01

A rapid and sensitive analytical method was developed to detect pathogenic bacteria which combined magnetic enrichment, fluorescence labeling with polyethylene glycol (PEG) magnetophoretic chromatography. As pathogenic bacteria usually exist in complex matrixes at low concentration, an efficient enrichment is essential for diagnosis. In order to capture series types of pathogenic bacteria in samples, amino-modified magnetic nanoparticles (Fe 3 O 4 @SiO 2 -NH 2 ) were prepared for efficient enrichment by the electrostatic interaction with pathogenic bacteria. It was shown that the capture efficiency reached up to 95.4% for Escherichia coli (E. coli). Furthermore, quantitative analysis of the bacteria was achieved by using acridine orange (AO) as a fluorescence probe for the captured E. coli due to its ability of staining series types of bacteria and rapid labeling. In order to remove the free magnetic nanoparticles and redundant fluorescent reagent, the labeled suspension was poured into a PEG separation column and was separated by applying an external magnetic field. The presence of 100 cfu mL -1 E. coli could be detected for semi-quantitative analysis by observing the separation column with the naked eye, and the concentration could be further evaluated by fluorescence detection. All the above processes were finished within 80 min. It was demonstrated that a good linear relationship existed between the fluorescence intensity and the concentration of E. coli ranging from 10 2 to 10 6  cfu mL -1 , with a detection limit of 100 cfu mL -1 when E. coli acted as target bacteria. The recovery rate of E. coli was 93.6∼102.0% in tap water and cooked meat samples, and the RSD was lower than 7% (n = 6); the result coincided with the conventional plate count method. Graphical abstract ᅟ.

Corn cob biochar increases soil culturable bacterial abundance without enhancing their capacities in utilizing carbon sources in Biolog Eco-plates

Institute of Scientific and Technical Information of China (English)

JIANG Lin-lin; HAN Guang-ming; LAN Yu; LIU Sai-nan; GAO Ji-ping; YANG Xu; MENG Jun; CHEN Wen-fu

2017-01-01

Biochar has been shown to influence soil microbial communities in terms of their abundance and diversity.However,the relationship among microbial abundance,structure and C metabolic traits is not well studied under biochar application.Here it was hypothesized that the addition of biochar with intrinsic properties (i.e.,porous structure) could affect the proliferation of culturable microbes and the genetic structure of soil bacterial communities.In the meantime,the presence of available organic carbon in biochar may influence the C utilization capacities of microbial community in Biolog Eco-plates.A pot experiment was conducted with differenct biochar application (BC) rates:control (0 t ha-1),BC1 (20 t ha-1) and BC2 (40 t ha-1).Culturable microorganisms were enumerated via the plate counting method.Bacterial diversity was examined using denaturing gradient gel electrophoresis (DGGE).Microbial capacity in using C sources was assessed using Biolog Eco-plates.The addition of biochar stimulated the growth of actinomyces and bacteria,especially the ammonifying bacteria and azotobacteria,but had no significant effect on fungi proliferation.The phylogenetic distribution of the operational taxonomic units could be divided into the following groups with the biochar addition:Firmicutes,Acidobacteria,Gemmatimonadetes,Actinobacteria,Cyanobacteria and α-,β-,γ-and δ-Proteobacteria (average similarity ＞95％).Biochar application had a higher capacity utilization for L-asparagine,Tween 80,D-mannitol,L-serine,γ-hydroxybutyric acid,N-acetyI-D-glucosamine,glycogen,itaconic acid,glycyl-L-glutamic acid,α-ketobutyricacid and putrescine,whereas it had received decreased capacities in using the other 20 carbon sources in Biolog Eco-plates.Redundancy analysis (RDA) revealed that the physico-chemical properties,indices of bacterial diversity,and C metabolic traits were positively correlated with the appearance of novel sequences under BC2 treatment.Our study indicates that the

Fitting a distribution to miccrobial counts: making sense of zeros

DEFF Research Database (Denmark)

Ribeiro Duarte, Ana Sofia; Stockmarr, Anders; Nauta, Maarten

and standard deviation) and the prevalence of contaminated food units (one minus the proportion of “true zeros”) from a set of microbial counts. By running the model with in silico generated concentration and count data, we could evaluate the performance of this method in terms of estimation of the three......Non-detects or left-censored results are inherent to the traditional methods of microbial enumeration in foods. Typically, a low concentration of microorganisms in a food unit goes undetected in plate counts or most probable number (MPN) counts, and produces “artificial zeros”. However......, these “artificial zeros” are only a share of the total number of zero counts resulting from a sample, as their number adds up to the number of “true zeros” resulting from uncontaminated units. In the process of fitting a probability distribution to microbial counts, “artificial” and “true” zeros are usually...

Enumeration and biomass estimation of planktonic bacteria and viruses by transmission electron microscopy

International Nuclear Information System (INIS)

Borsheim, K.Y.; Bratbak, G.; Heldal, M.

1990-01-01

Bacteria and virus particles were harvested from water samples by ultracentrifugation directly onto Formvar-coated electron microscopy grids and counted in a transmission electron microscope. With this technique, we have counted and sized bacteria and viruses in marine water samples and during laboratory incubations. By X-ray microanalysis, we could determine the elemental composition and dry-matter content of individual bacteria. The dry weight/volume ratio for the bacteria was 600 fg of dry weight microns-3. The potassium content of the bacteria was normal compared with previous estimates from other bacterial assemblages; thus, this harvesting procedure did not disrupt the bacterial cells. Virus particles were, by an order of magnitude, more abundant than bacteria in marine coastal waters. During the first 5 to 7 days of incubation, the total number of viruses increased exponentially at a rate of 0.4 day-1 and thereafter declined. The high proliferation rate suggests that viral parasitism may affect mortality of bacteria in aquatic environments

Isolation and characterization of bacteria flora from dumpsites in ...

African Journals Online (AJOL)

The bacterial load count for the various dumpsites and the adjacent control sites was counted using mise and misra serial dilution method. Aerobic and facultative anaerobic bacteria were isolated from dump site samples using five different culture media. Phenotypic test was assayed using double disk method to check for ...

Processing ruminal ingesta to release bacteria attached to feed ...

African Journals Online (AJOL)

A comparison was made of different methods of processing ingesta to release bacteria attached to solid particles, prior to making viable counts. Initially processing was performed under a stream of anaerobic gas and counts were made using the roll tube technique. Later, processing was done in an anaerobic cabinet and ...

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African Journals Online (AJOL)

DR. AMINU

2008-12-01

Dec 1, 2008 ... The counts from raw meat were, aerobic plate count (APC) = 1.73 E8 cfu/g, and fungal counts (FC) = 2.02 E7 cfu/g, ... Key words: Hazard and Critical Control Point, Bacterial count, Fungal count, Pathogenic bacteria, balangu, Kano. .... Does a control measure for the hazard exist at this step. Is control at this ...

Use of pliable bags in liquid scintillation counting

International Nuclear Information System (INIS)

Simonnet, G.; Jacquet, M.A.; Sharif, A.; Engler, R.

1981-01-01

Pliable plastic bags have been used to replace glass or plastic vials for liquid scintillation counting. The two major advantages of this method are the lower cost of the plastic bags and the fact that, per sample, the radioactive waste is significantly reduced. The following parameters have been checked: the impermeability of the bags to various scintillator mixtures and the fact that neither the irregular shape of the bags nor their position in the counting chamber had any effect on the results of the counting. The latter was also constant with time, at least over a period of 10 days. The technique has been used to count the radioactivity of 3 H-DNA precipitates prepared from bacteria and lymphocytes and deposited on filters impregnated with only 200 μl scintillator. It is a method that can be applied to the counting of any samples deposited on filters and insoluble in scintillator. (author)

BACTERIOLOGICAL PROPERTIES OF MARINE WATER IN ADRIATIC FISH FARMS: ENUMERATION OF HETEROTROPHIC BACTERIA

Directory of Open Access Journals (Sweden)

Emin Teskeredžić

2012-12-01

Full Text Available Aquaculture is currently one of the fastest growing food production sectors in the world. Increase in nutrients and organic wastes lead to general deterioration of water quality. The problem of water quality is associated with both physical and chemical factors, as well as microbiological water quality. Heterotrophic bacteria play an important role in the process of decomposition of organic matter in water environment and indicate eutrophication process. Here we present our experience and knowledge on bacterial properties of marine water in the Adriatic fish farms with European sea bass (Dicentrarchus labrax L., 1758, with an emphasis on enumeration of heterotrophic bacteria in marine water. We applied two temperatures of incubation, as well as two methods for enumeration of heterotrophic bacteria: substrate SimPlate® test and spread plate method on conventional artificial media (Marine agar and Tryptic Soy agar with added NaCl. The results of analysis of bacteriological properties of marine water in the Adriatic fish farms showed that enumeration of heterotrophic bacteria in marine water depends on the applied incubation temperature and media for enumeration. At the same time, the incubation temperature of 22C favours more intense growth of marine heterotrophic bacteria, whereas a SimPlate test gives higher values of heterotrophic bacteria. Volatile values of heterotrophic bacteria during this research indicate a possible deterioration of microbiological water quality in the Adriatic fish farms and a need for regular monitoring of marine water quality.

Undercooling and nodule count in thin walled ductile iron castings

DEFF Research Database (Denmark)

Pedersen, Karl Martin; Tiedje, Niels Skat

2007-01-01

Casting experiments have been performed with eutectic and hypereutectic castings with plate thicknesses from 2 to 8 mm involving both temperature measurements during solidification and microstructural examination afterwards. The nodule count was the same for the eutectic and hypereutectic casting...

Ozone Technology for Pathogenic Bacteria of Shrimp (Vibrio sp.) Disinfection

Science.gov (United States)

Wulansarie, Ria; Dyah Pita Rengga, Wara; Rustamadji

2018-03-01

One of important marine commodities in Indonesia, shrimps are susceptible with Vibrio sp bacteria infection. That infection must be cleared. One of the technologies for disinfecting Vibrio sp. is ozone technology. In this research, Vibrio sp. is a pathogenic bacterium which infects Penaeus vannamei. Ozone technology is applied for threatening Vibrio sp. In this research, ozonation was performed in different pH. Those are neutral, acid (pH=4), and base (pH=9). The sample was water from shrimp embankment from Balai Besar Perikanan Budidaya Air Payau (BBPBAP) located in Jepara. That water was the habitat of Penaeus vannamei shrimp. The brand of ozonator used in this research was “AQUATIC”. The used ozonator in this research had 0,0325 g/hour concentration. The flow rate of sample used in this research was 2 L/minute. The ozonation process was performed in continuous system. A tank, pipe, pump, which was connected with microfilter, flowmeter and ozone generator were the main tools in this research. It used flowmeter and valve to set the flow rate scalable as desired. The first step was the insert of 5 L sample into the receptacle. Then, by using a pump, a sample supplied to the microfilter to be filtered and passed into the flow meter. The flow rate was set to 2 LPM. Furthermore, gas from ozonator passed to the flow for the disinfection of bacteria and then was recycled to the tank and the process run continuously. Samples of the results of ozonation were taken periodically from time 0, 3, 7, 12, 18, 24 to 30 minutes. The samples of the research were analyzed using Total Plate Count (TPC) test in BBPBAP Jepara to determine the number of Vibrio sp. bacteria. The result of this research was the optimal condition for pathogenic bacteria of shrimp (Vibrio sp.) ozonation was in neutral condition.

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

Science.gov (United States)

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.

Identifying the major bacteria causing intramammary infections in individual milk samples of sheep and goats using traditional bacteria culturing and real-time polymerase chain reaction.

Science.gov (United States)

Rovai, M; Caja, G; Salama, A A K; Jubert, A; Lázaro, B; Lázaro, M; Leitner, G

2014-09-01

Use of DNA-based methods, such as real-time PCR, has increased the sensitivity and shortened the time for bacterial identification, compared with traditional bacteriology; however, results should be interpreted carefully because a positive PCR result does not necessarily mean that an infection exists. One hundred eight lactating dairy ewes (56 Manchega and 52 Lacaune) and 24 Murciano-Granadina dairy goats were used for identifying the main bacteria causing intramammary infections (IMI) using traditional bacterial culturing and real-time PCR and their effects on milk performance. Udder-half milk samples were taken for bacterial culturing and somatic cell count (SCC) 3 times throughout lactation. Intramammary infections were assessed based on bacteria isolated in ≥2 samplings accompanied by increased SCC. Prevalence of subclinical IMI was 42.9% in Manchega and 50.0% in Lacaune ewes and 41.7% in goats, with the estimated milk yield loss being 13.1, 17.9, and 18.0%, respectively. According to bacteriology results, 87% of the identified single bacteria species (with more than 3 colonies/plate) or culture-negative growth were identical throughout samplings, which agreed 98.9% with the PCR results. Nevertheless, the study emphasized that 1 sampling may not be sufficient to determine IMI and, therefore, other inflammatory responses such as increased SCC should be monitored to identify true infections. Moreover, when PCR methodology is used, aseptic and precise milk sampling procedures are key for avoiding false-positive amplifications. In conclusion, both PCR and bacterial culture methods proved to have similar accuracy for identifying infective bacteria in sheep and goats. The final choice will depend on their response time and cost analysis, according to the requirements and farm management strategy. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Assessment of Antibiotic Resistant Commensal Bacteria in Food

Science.gov (United States)

2006-01-01

mold Penicillium was able to inhibit the growth of some bacteria. (37). In 1928, a Brittish physician Alexander Fleming observed the similar...phenomenon. One of his bacterial plate cultures was contaminated with the blue-green mold Penicillium , and the bacterial colonies in close approximation to...sliced chicken lunchmeat, and the strain was identified as Pseudomonas sp . ART bacteria were isolated sporadically in 32 lunchmeat, which is

Relationship between aerobic bacteria, salmonellae and Campylobacter on broiler carcasses.

Science.gov (United States)

Cason, J A; Bailey, J S; Stern, N J; Whittemore, A D; Cox, N A

1997-07-01

Broiler carcasses were removed from commercial processing lines immediately after defeathering, before chilling, and after chilling to determine whether any relationship exists between aerobic bacteria and the human enteropathogens salmonellae and Campylobacter. In two experiments, a whole carcass rinse procedure was used to sample 30 carcasses after defeathering, 90 carcasses before chilling, and 90 carcasses after chilling, for a total of 210 different carcasses. Aerobic bacteria and Campylobacter spp. were enumerated and the incidence of salmonellae was determined. Salmonellae and Campylobacter incidences were 20 and 94%, respectively, for all carcasses sampled. After picking, neither salmonellae-positive nor Campylobacter-positive carcasses had mean aerobic most probable number (MPN) values that were different from carcasses negative for those organisms. Immediately before chilling, aerobic and Campylobacter counts were 7.12 and 5.33 log10 cfu per carcass, respectively. Immersion chilling reduced aerobic counts by approximately 1.8 log and Campylobacter by 1.5 log, with no change in salmonellae-positive carcasses. There was no difference in aerobic or Campylobacter counts between carcasses that were positive or negative for salmonellae at any of the sampling locations, nor was any correlation found between levels of aerobic organisms and Campylobacter. Carcasses with aerobic counts above the mean or more than one standard deviation above the mean also failed to show any correlation. Discriminant analysis indicated error rates as high as 50% when numbers of aerobic bacteria were used to predict incidence of salmonellae or Campylobacter on individual carcasses. Aerobic bacteria are not suitable as index organisms for salmonellae or Campylobacter on broiler carcasses.

Comparison of bacteria populations in clean and recycled sand used for bedding in dairy facilities.

Science.gov (United States)

Kristula, M A; Rogers, W; Hogan, J S; Sabo, M

2005-12-01

Bedding samples were collected twice from commercial dairy free-stall facilities that used recycled sand and clean sand in both the summer and winter. Collection began on the day sand was taken from the pile (d 0) and placed in the free stalls, and continued for 5 to 7 additional days. The number of colonies per gram of bedding of gram-negative bacteria, coliforms, Streptococcus spp., and Klebsiella spp. were estimated for each sand sample as well as amounts of dry and organic matter. Clean sand (CS) and recycled sand (RS) had the same bacterial counts when compared at any sampling time. The mean counts of bacterial populations did vary over the course of the study in both CS and RS. There was a significant increase in bacterial counts from d 0 to d 1 for gram-negative bacteria, coliforms, and Streptococcus spp. in both winter and summer. Counts of gram-negative bacteria, coliforms, Klebsiella spp., and Streptococcus spp. did not differ from d 1 to 7 in the winter. Total counts of gram-negative bacteria did not differ from d 1 to 7 in the summer. On d 1 in the summer, coliform counts were lower than at d 5 to 7, and Klebsiella spp. counts were lower than on d 3 to 7. Streptococcus spp. counts were high on d 1 and were constant through d 7 in both winter and summer trials. The number of coliform and Klebsiella spp. in both CS and RS was below the threshold thought to cause mastitis during the sampling times. The number of Streptococcus spp. was high in both CS and RS during the sampling periods. Other management factors need to be identified to decrease the number of Streptococcus spp. in bedding. Recycled sand had a higher organic matter and lower dry matter compared with CS in winter and summer. The results for this study were obtained from multiple herd comparisons, and herd was a significant effect suggesting that different management systems influence the number and types of bacteria in both CS and RS.

Experimental Study for Automatic Colony Counting System Based Onimage Processing

Science.gov (United States)

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.

Culture dependent bacteria in commercial fishes: Qualitative assessment and molecular identification using 16S rRNA gene sequencing

KAUST Repository

Mannalamkunnath Alikunhi, Nabeel; Batang, Zenon B.; AlJahdali, Haitham A.; Aziz, Mohammed A.M.; Al-Suwailem, Abdulaziz M.

2016-01-01

Fish contaminations have been extensively investigated in Saudi coasts, but studies pertaining to bacterial pathogens are meager. We conducted qualitative assessment and molecular identification of culture dependent bacteria in 13 fish species collected from three fishing sites and a local fish market in Jeddah, Saudi Arabia. The bacterial counts of gills, skin, gut and muscle were examined on agar plates of Macconkey’s (Mac), Eosin methylene blue (EMB) and Thiosulfate Citrate Bile Salts (TCBS) culture media. Bacterial counts exhibited interspecific, locational and behavioral differences. Mugil cephalus exhibited higher counts on TCBS (all body-parts), Mac (gills, muscle and gut) and EMB (gills and muscle). Samples of Area I were with higher counts, concurrent to seawater and sediment samples, revealing the influence of residing environment on fish contamination. Among feeding habits, detritivorous fish harbored higher bacterial counts, while carnivorous group accounted for lesser counts. Counts were higher in skin of fish obtained from market compared to field samples, revealing market as a major source of contamination. Bacterial counts of skin were positively correlated with other body-parts indicating influence of surface bacterial biota in overall quality of fish. Hence, hygienic practices and proper storage facilities in the Jeddah fish market is recommended to prevent adverse effect of food-borne illness in consumers. Rahnella aquatilis (Enterobacteriaceae) and Photobacterium damselae (Vibrionaceae) were among the dominant species identified from fish muscle samples using Sanger sequencing of 16S rRNA. This bacterial species are established human pathogens capable of causing foodborne illness with severe antibiotic resistance. Opportunistic pathogens such as Hafnia sp. (Enterobacteriaceae) and Pseudomonas stutzeri (Pseudomonadaceae) were also identified from fish muscle. These findings indicate bacterial contamination risk in commonly consumed fish of

Culture dependent bacteria in commercial fishes: Qualitative assessment and molecular identification using 16S rRNA gene sequencing

KAUST Repository

Alikunhi, Nabeel M.

2016-05-27

Fish contaminations have been extensively investigated in Saudi coasts, but studies pertaining to bacterial pathogens are meager. We conducted qualitative assessment and molecular identification of culture dependent bacteria in 13 fish species collected from three fishing sites and a local fish market in Jeddah, Saudi Arabia. The bacterial counts of gills, skin, gut and muscle were examined on agar plates of Macconkey’s (Mac), Eosin methylene blue (EMB) and Thiosulfate Citrate Bile Salts (TCBS) culture media. Bacterial counts exhibited interspecific, locational and behavioral differences. Mugil cephalus exhibited higher counts on TCBS (all body-parts), Mac (gills, muscle and gut) and EMB (gills and muscle). Samples of Area I were with higher counts, concurrent to seawater and sediment samples, revealing the influence of residing environment on fish contamination. Among feeding habits, detritivorous fish harbored higher bacterial counts, while carnivorous group accounted for lesser counts. Counts were higher in skin of fish obtained from market compared to field samples, revealing market as a major source of contamination. Bacterial counts of skin were positively correlated with other body-parts indicating influence of surface bacterial biota in overall quality of fish. Hence, hygienic practices and proper storage facilities in the Jeddah fish market is recommended to prevent adverse effect of food-borne illness in consumers. Rahnella aquatilis (Enterobacteriaceae) and Photobacterium damselae (Vibrionaceae) were among the dominant species identified from fish muscle samples using Sanger sequencing of 16S rRNA. This bacterial species are established human pathogens capable of causing foodborne illness with severe antibiotic resistance. Opportunistic pathogens such as Hafnia sp. (Enterobacteriaceae) and Pseudomonas stutzeri (Pseudomonadaceae) were also identified from fish muscle. These findings indicate bacterial contamination risk in commonly consumed fish of

Effects of supplementing lactic acid bacteria on fecal microbiota ...

African Journals Online (AJOL)

Results: The results indicated that Lactobacillus plantarum strain L.p X3-2B increased fecal lactic acid bacteria(LAB) and Bifidobacterium while resisting the growth of harmful bacteria. Viable counts of LAB and Bifidobacterium reached 8 log cfu/mL after feeding for 14 days. Fecal pH in the control group was high in ...

Bio-protective potential of lactic acid bacteria: Effect of and on changes of the microbial community in vacuum-packaged chilled beef

Directory of Open Access Journals (Sweden)

Yimin Zhang

2018-04-01

Full Text Available Objective This study was to determine the bacterial diversity and monitor the community dynamic changes during storage of vacuum-packaged sliced raw beef as affected by Lactobacillus sakei and Lactobacillus curvatus. Methods L. sakei and L. curvatus were separately incubated in vacuumed-packaged raw beef as bio-protective cultures to inhibit the naturally contaminating microbial load. Dynamic changes of the microbial diversity of inoculated or non-inoculated (control samples were monitored at 4°C for 0 to 38 days, using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE. Results The DGGE profiles of DNA directly extracted from non-inoculated control samples highlighted the order of appearance of spoilage bacteria during storage, showing that Enterbacteriaceae and Pseudomonas fragi emerged early, then Brochothrix thermosphacta shared the dominant position, and finally, Pseudomonas putida showed up became predominant. Compared with control, the inoculation of either L. sakei or L. curvatus significantly lowered the complexity of microbial diversity and inhibited the growth of spoilage bacteria (p<0.05. Interestingly, we also found that the dominant position of L. curvatus was replaced by indigenous L. sakei after 13 d for L. curvatus-inoculated samples. Plate counts on selective agars further showed that inoculation with L. sakei or L. curvatus obviously reduced the viable counts of Enterbacteraceae, Pseudomonas spp. and B. thermosphacta during later storage (p< 0.05, with L. sakei exerting greater inhibitory effect. Inoculation with both bio-protective cultures also significantly decreased the total volatile basic nitrogen values of stored samples (p<0.05. Conclusion Taken together, the results proved the benefits of inoculation with lactic acid bacteria especially L. sakei as a potential way to inhibit growth of spoilage-related bacteria and improve the shelf life of vacuum-packaged raw beef.

Distribution and species composition of planktonic luminous bacteria in the Arabian Sea

Digital Repository Service at National Institute of Oceanography (India)

Ramaiah, N.; Chandramohan, D.

Distribution of the total viable heterotrophic bacteria and the luminous bacteria in the neretic and oceanic waters of the west coast of India was studied. Counts of viable heterotrophs fluctuated widely, generally with a decrease in their number...

Cellular viability of Saccharomyces cerevisiae cultivated in association with contaminates bacteria of alcoholic fermentation;Viabilidade celular de Saccharomyces cerevisiae cultivada em associacao com bacterias contaminantes da fermentacao alcoolica

Energy Technology Data Exchange (ETDEWEB)

Nobre, Thais de Paula

2005-07-01

The aim of this work was to study the influence of the bacteria Bacillus and Lactobacillus, as well as their metabolic products, in reduction of cellular viability of Saccharomyces cerevisiae, when in mixed culture of yeast and active and treated bacteria. Also was to evaluated an alternative medium (MCC) for the cultivation of bacteria and yeast, constituted of sugarcane juice diluted to 5 deg Brix and supplemented with yeast extract and peptone. The bacteria Bacillus subtilis, Bacillus coagulans, Bacillus stearothermophilus, Lactobacillus fermentum and Lactobacillus plantarum were cultivated in association with yeast Saccharomyces cerevisiae (strain Y-904) for 72 h on 32 deg C, under agitation. The cellular viability, budding rate and population of S. cerevisiae, the total acidity, volatile acidity and pH of culture were determined from 0, 24, 48 e 72 h of mixed culture. Also were determined the initial and final of microorganism population across the pour plate method, in traditional culture medium (PCA for Bacillus, MRS-agar for Lactobacillus and YEPD-agar for yeast S. cerevisiae) and in medium constituted of sugarcane juice. The bacteria cultures were treated by heat sterilization (120 deg C for 20 minutes), antibacterial agent (Kamoran HJ in concentration 3,0 ppm) or irradiation (radiation gamma, with doses of 5,0 kGy for Lactobacillus and 15,0 kGy for Bacillus). The results of the present research showed that just the culture mediums more acids (with higher concentrations of total and volatile acidity, and smaller values of pH), contaminated with active bacteria L. fermentum and B. subtilis, caused reduction on yeast cellular viability. Except the bacteria B. subtilis treated with radiation, the others bacteria treated by different procedures (heat, radiation e antibacterial) did not cause reduction on yeast cellular viability and population, indicating that the isolated presence of the cellular metabolic of theses bacteria was not enough to reduce the

A novel, optical, on-line bacteria sensor for monitoring drinking water quality.

Science.gov (United States)

Højris, Bo; Christensen, Sarah Christine Boesgaard; Albrechtsen, Hans-Jørgen; Smith, Christian; Dahlqvist, Mathis

2016-04-04

Today, microbial drinking water quality is monitored through either time-consuming laboratory methods or indirect on-line measurements. Results are thus either delayed or insufficient to support proactive action. A novel, optical, on-line bacteria sensor with a 10-minute time resolution has been developed. The sensor is based on 3D image recognition, and the obtained pictures are analyzed with algorithms considering 59 quantified image parameters. The sensor counts individual suspended particles and classifies them as either bacteria or abiotic particles. The technology is capable of distinguishing and quantifying bacteria and particles in pure and mixed suspensions, and the quantification correlates with total bacterial counts. Several field applications have demonstrated that the technology can monitor changes in the concentration of bacteria, and is thus well suited for rapid detection of critical conditions such as pollution events in drinking water.

OBTAINING OF PROTEIC BIOMASS BY CULTIVATION OF LACTIC ACID BACTERIA ON GRAPE MARC DIFFUSION SOLUTION

Directory of Open Access Journals (Sweden)

Marian BUTU

2013-08-01

Full Text Available In this article are presented the researches made in order to obtain protein biomass with the aid of lactic bacteria grown on an economically medium, achieved by using secondary products from the winery: marc and wine yeast. Therefore, there were cultivated two strains of Lactobacillus sp. on five different growth medium. The protein biosynthesis and evolution of lactic fermentation were monitored by determining the optical density (OD of the culture at a wavelength λ = 600 nm and by counting the colony forming units (CFU by serial dilutions and seeding on plates and by determination of lactic acid obtained. The results showed that the fermentation medium represented by diffusion solution of the marc, enriched with peptone is economically profitable compared to other culture media containing peptone, yeast extract, glucose, minerals, amino acids and vitamins presented in the literature.

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.

Science.gov (United States)

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.

Immunological techniques as tools to characterize the subsurface microbial community at a trichloroethylene contaminated site

Energy Technology Data Exchange (ETDEWEB)

Fliermans, C.B.; Dougherty, J.M.; Franck, M.M.; McKinzey, P.C.; Hazen, T.C.

1992-01-01

Effective in situ bioremediation strategies require an understanding of the effects pollutants and remediation techniques have on subsurface microbial communities. Therefore, detailed characterization of a site's microbial communities is important. Subsurface sediment borings and water samples were collected from a trichloroethylene (TCE) contaminated site, before and after horizontal well in situ air stripping and bioventing, as well as during methane injection for stimulation of methane-utilizing microorganisms. Subsamples were processed for heterotrophic plate counts, acridine orange direct counts (AODC), community diversity, direct fluorescent antibodies (DFA) enumeration for several nitrogen-transforming bacteria, and Biolog [reg sign] evaluation of enzyme activity in collected water samples. Plate counts were higher in near-surface depths than in the vadose zone sediment samples. During the in situ air stripping and bioventing, counts increased at or near the saturated zone, remained elevated throughout the aquifer, but did not change significantly after the air stripping. Sporadic increases in plate counts at different depths as well as increased diversity appeared to be linked to differing lithologies. AODCs were orders of magnitude higher than plate counts and remained relatively constant with depth except for slight increases near the surface depths and the capillary fringe. Nitrogen-transforming bacteria, as measured by serospecific DFA, were greatly affected both by the in situ air stripping and the methane injection. Biolog[reg sign] activity appeared to increase with subsurface stimulation both by air and methane. The complexity of subsurface systems makes the use of selective monitoring tools imperative.

Immunological techniques as tools to characterize the subsurface microbial community at a trichloroethylene contaminated site

Energy Technology Data Exchange (ETDEWEB)

Fliermans, C.B.; Dougherty, J.M.; Franck, M.M.; McKinzey, P.C.; Hazen, T.C.

1992-12-31

Effective in situ bioremediation strategies require an understanding of the effects pollutants and remediation techniques have on subsurface microbial communities. Therefore, detailed characterization of a site`s microbial communities is important. Subsurface sediment borings and water samples were collected from a trichloroethylene (TCE) contaminated site, before and after horizontal well in situ air stripping and bioventing, as well as during methane injection for stimulation of methane-utilizing microorganisms. Subsamples were processed for heterotrophic plate counts, acridine orange direct counts (AODC), community diversity, direct fluorescent antibodies (DFA) enumeration for several nitrogen-transforming bacteria, and Biolog {reg_sign} evaluation of enzyme activity in collected water samples. Plate counts were higher in near-surface depths than in the vadose zone sediment samples. During the in situ air stripping and bioventing, counts increased at or near the saturated zone, remained elevated throughout the aquifer, but did not change significantly after the air stripping. Sporadic increases in plate counts at different depths as well as increased diversity appeared to be linked to differing lithologies. AODCs were orders of magnitude higher than plate counts and remained relatively constant with depth except for slight increases near the surface depths and the capillary fringe. Nitrogen-transforming bacteria, as measured by serospecific DFA, were greatly affected both by the in situ air stripping and the methane injection. Biolog{reg_sign} activity appeared to increase with subsurface stimulation both by air and methane. The complexity of subsurface systems makes the use of selective monitoring tools imperative.

A flow cytometric technique for quantification and differentiation of bacteria in bulk tank milk

DEFF Research Database (Denmark)

Holm, C.; Mathiasen, T.; Jespersen, Lene

2004-01-01

were defined: region 1 includes bacteria mainly associated with poor hygiene, region 2 includes psychrotrophic hygiene bacteria and region 3 includes bacteria mainly related to mastitis. The ability of the flow cytometric technique to predict the main cause of elevated bacterial counts on routine...

Evaluation of changes in periodontal bacteria in healthy dogs over 6 months using quantitative real-time PCR.

Science.gov (United States)

Maruyama, N; Mori, A; Shono, S; Oda, H; Sako, T

2018-03-01

Porphyromonas gulae, Tannerella forsythia and Campylobacter rectus are considered dominant periodontal pathogens in dogs. Recently, quantitative real-time PCR (qRT-PCR) methods have been used for absolute quantitative determination of oral bacterial counts. The purpose of the present study was to establish a standardized qRT-PCR procedure to quantify bacterial counts of the three target periodontal bacteria (P. gulae, T. forsythia and C. rectus). Copy numbers of the three target periodontal bacteria were evaluated in 26 healthy dogs. Then, changes in bacterial counts of the three target periodontal bacteria were evaluated for 24 weeks in 7 healthy dogs after periodontal scaling. Analytical evaluation of each self-designed primer indicated acceptable analytical imprecision. All 26 healthy dogs were found to be positive for P. gulae, T. forsythia and C. rectus. Median total bacterial counts (copies/ng) of each target genes were 385.612 for P. gulae, 25.109 for T. forsythia and 5.771 for C. rectus. Significant differences were observed between the copy numbers of the three target periodontal bacteria. Periodontal scaling reduced median copy numbers of the three target periodontal bacteria in 7 healthy dogs. However, after periodontal scaling, copy numbers of all three periodontal bacteria significantly increased over time (pperiodontal bacteria in dogs. Furthermore, the present study has revealed that qRT-PCR method can be considered as a new objective evaluation system for canine periodontal disease. Copyright© by the Polish Academy of Sciences.

Bacteria associated with cysts of the soybean cyst nematode (Heterodera glycines).

Science.gov (United States)

Nour, Sarah M; Lawrence, John R; Zhu, Hong; Swerhone, George D W; Welsh, Martha; Welacky, Tom W; Topp, Edward

2003-01-01

The soybean cyst nematode (SCN), Heterodera glycines, causes economically significant damage to soybeans (Glycine max) in many parts of the world. The cysts of this nematode can remain quiescent in soils for many years as a reservoir of infection for future crops. To investigate bacterial communities associated with SCN cysts, cysts were obtained from eight SCN-infested farms in southern Ontario, Canada, and analyzed by culture-dependent and -independent means. Confocal laser scanning microscopy observations of cyst contents revealed a microbial flora located on the cyst exterior, within a polymer plug region and within the cyst. Microscopic counts using 5-(4,6-dichlorotriazine-2-yl)aminofluorescein staining and in situ hybridization (EUB 338) indicated that the cysts contained (2.6 +/- 0.5) x 10(5) bacteria (mean +/- standard deviation) with various cellular morphologies. Filamentous fungi were also observed. Live-dead staining indicated that the majority of cyst bacteria were viable. The probe Nile red also bound to the interior polymer, indicating that it is lipid rich in nature. Bacterial community profiles determined by denaturing gradient gel electrophoresis analysis were simple in composition. Bands shared by all eight samples included the actinobacterium genera Actinomadura and STREPTOMYCES: A collection of 290 bacteria were obtained by plating macerated surface-sterilized cysts onto nutrient broth yeast extract agar or on actinomycete medium. These were clustered into groups of siblings by repetitive extragenic palindromic PCR fingerprinting, and representative isolates were tentatively identified on the basis of 16S rRNA gene sequence. Thirty phylotypes were detected, with the collection dominated by Lysobacter and Variovorax spp. This study has revealed the cysts of this important plant pathogen to be rich in a variety of bacteria, some of which could presumably play a role in the ecology of SCN or have potential as biocontrol agents.

Growth of legionella and other heterotrophic bacteria in a circulating cooling water system exposed to ultraviolet irradiation

International Nuclear Information System (INIS)

Kusnetsov, J.M.; Miettinen, I.T.; Martikainen, P.J.; Keskitalo, P.J.; Ahonen, H.E.; Tulkki, A.I.

1994-01-01

The effect of ultraviolet irradiation on the growth and occurrence of legionella and other heterotrophic bacteria in a circulating cooling water system was studied. Water of the reservoir was circulated once in 28 h through a side-stream open channel u.v. radiator consisting of two lamps. Viable counts of legionellas and heterotrophic bacteria in water immediately after the u.v. treatment were 0-12 and 0.7-1.2% of those in the reservoir, respectively. U.v. irradiation increased the concentration of easily assimilable organic carbon. In the u.v. irradiated water samples incubated in the laboratory the viable counts of heterotropic bacteria reached the counts in reservoir water within 5 d. The increase in viable counts was mainly due to reactivation of bacterial cells damaged by u.v. light, not because of bacterial multiplication. Despite u.v. irradiation the bacterial numbers in the reservoir water, including legionellas, did not decrease during the experimental period of 33 d. The main growth of bacteria in the reservoir occurred in biofilm and sediment, which were never exposed to u.v. irradiation. (Author)

Assessment of the application of an automated electronic milk analyzer for the enumeration of total bacteria in raw goat milk.

Science.gov (United States)

Ramsahoi, L; Gao, A; Fabri, M; Odumeru, J A

2011-07-01

Automated electronic milk analyzers for rapid enumeration of total bacteria counts (TBC) are widely used for raw milk testing by many analytical laboratories worldwide. In Ontario, Canada, Bactoscan flow cytometry (BsnFC; Foss Electric, Hillerød, Denmark) is the official anchor method for TBC in raw cow milk. Penalties are levied at the BsnFC equivalent level of 50,000 cfu/mL, the standard plate count (SPC) regulatory limit. This study was conducted to assess the BsnFC for TBC in raw goat milk, to determine the mathematical relationship between the SPC and BsnFC methods, and to identify probable reasons for the difference in the SPC:BsnFC equivalents for goat and cow milks. Test procedures were conducted according to International Dairy Federation Bulletin guidelines. Approximately 115 farm bulk tank milk samples per month were tested for inhibitor residues, SPC, BsnFC, psychrotrophic bacteria count, composition (fat, protein, lactose, lactose and other solids, and freezing point), and somatic cell count from March 2009 to February 2010. Data analysis of the results for the samples tested indicated that the BsnFC method would be a good alternative to the SPC method, providing accurate and more precise results with a faster turnaround time. Although a linear regression model showed good correlation and prediction, tests for linearity indicated that the relationship was linear only beyond log 4.1 SPC. The logistic growth curve best modeled the relationship between the SPC and BsnFC for the entire sample population. The BsnFC equivalent to the SPC 50,000 cfu/mL regulatory limit was estimated to be 321,000 individual bacteria count (ibc)/mL. This estimate differs considerably from the BsnFC equivalent for cow milk (121,000 ibc/mL). Because of the low frequency of bulk tank milk pickups at goat farms, 78.5% of the samples had their oldest milking in the tank to be 6.5 to 9.0 d old when tested, compared with the cow milk samples, which had their oldest milking at 4 d

Endophytes diversity of bacteria associated with roots of colosuana (bothriochloa pertusa) pasture in three locations of Sucre Department, Colombia

International Nuclear Information System (INIS)

Perez C, Alexander; Rojas S, Johanna; Fuentes C, Justo

2010-01-01

The objective of this study was to isolate endophytes diversity of culturable bacteria associated with grass roots colosuana Bothriochloa pertusa (L) a. camus in three localities of the department of Sucre, Colombia. endophytes bacterial diversity was performed by isolation of colonies on media culture. The population density was estimated by direct counting of colonies on plate and cultural characteristics of each morphotype were obtained by observation of each colony made. We determined the correlation diversity, population density and locations, using ANOVA and principal component analysis or simple correspondence, using the statistical program R, 2009 (4.5). 20 farms livestock were sampled by locality; it was observed the presence of various bacterial morphotypes endophytes. We found significant differences between diversity (morphotypes), population density (UFC.raiz -1 ) and locations. The diversity of bacterial endophytes represents only a small fraction of the total diversity present in nature; being little information we have of the presence of these microorganisms in specific agro-ecosystems, which is why this work becomes the first evidence of association between bacteria and roots of grass endophytes colosuana in Colombia.

Preparation of actinide targets by molecular plating for coulomb excitation studies at ATLAS

CERN Document Server

Greene, J P; Ahmad, I

1999-01-01

Molecular plating is now routinely used to prepare sources and targets of actinide elements. Although the technique is simple and fairly reproducible, because of the radioactive nature of the targets, it is very useful to record various parameters in the preparation process. At Argonne, approx 200 mu g/cm sup 2 thick targets of Pu and Cm were required for Coulomb Excitation (COULEX) studies with the Argonne-Notre Dame boron germanate (BGO) gamma-ray facility and later with the GAMMASPHERE. These targets were plated on 50 mg/cm sup 2 Au backings and were covered with 150 mu g/cm sup 2 Au foil. Targets of sup 2 sup 3 sup 9 Pu, sup 2 sup 4 sup 0 Pu, sup 2 sup 4 sup 2 Pu, sup 2 sup 4 sup 4 Pu and sup 2 sup 4 sup 8 Cm were prepared by dissolving the material in isopropyl alcohol and electroplating the actinide ions by applying 600 V. The amount of these materials on the target was determined by alpha particle counting and gamma-ray counting. Details of the molecular plating and counting will be discussed.

Culture-dependent bacteria in commercial fishes: Qualitative assessment and molecular identification using 16S rRNA gene sequencing

Directory of Open Access Journals (Sweden)

Nabeel M. Alikunhi

2017-09-01

Full Text Available Fish contamination has been extensively investigated along the Saudi coasts, but studies pertaining to bacterial pathogens are scarce. We conducted qualitative assessment and molecular identification of culture-dependent bacteria in 13 fish species from three coastal sites and a local fish market in Jeddah, Saudi Arabia. Bacterial counts of gills, skin, gut and muscle were examined on agar plates of Macconkey’s (Mac, Eosin Methylene Blue (EMB and Thiosulfate Citrate Bile Salts (TCBS culture media. Bacterial counts significantly differed between species, sources and feeding habits of examined fishes. Mugil cephalus exhibited higher counts on TCBS (all body parts, Mac (gills, muscle and gut and EMB (gills and muscle. Fishes from Area I had higher bacterial loads, coinciding with those in seawater and sediment from the same site, indicating direct association between habitat conditions and the levels of bacterial contamination. By feeding habit, detritivorous fish harbored higher counts than herbivorous and carnivorous species. Bacterial counts of skin were higher in fish from market than field sites, and positively correlated with other body parts indicating the relation of surface bacterial load on the overall quality of fish. Rahnella aquatilis (Enterobacteriaceae and Photobacterium damselae (Vibrionaceae were among the dominant species from fish muscle based on 16S rRNA sequencing. These species are known human pathogens capable of causing foodborne illness with severe antibiotic resistance. Opportunistic pathogens, e.g. Hafnia sp. (Enterobacteriaceae and Pseudomonas stutzeri (Pseudomonadaceae also occurred in fish muscle. The inclusion of bacterial contamination in future monitoring efforts is thus crucial.

Evaluation of PCR and DNA hybridization protocols for detection of viable enterotoxigenic Clostridium perfringens in irradiated beef

International Nuclear Information System (INIS)

Baez, L.A.; Juneja, V.K.; Thayer, D.W.; Sackitey, S.

1997-01-01

The sensitivity of DNA hybridization and polymerase chain reaction (PCR), was evaluated in irradiated cooked and raw beef samples. A membrane-based colony hybridization assay and a PCR protocol, both with specificity for the enterotoxin A gene of Clostridium perfringens, were compared with viable plate counts. The results of the colony hybridization procedure were in agreement with viable plate counts for detection and enumeration of enterotoxigenic C. perfringens. The PCR procedure combined a 4 h enrichment followed by a nucleic acid extraction step and assessed the amplification of 183 and 750 base pair enterotoxin gene targets. Detection of C. perfringens by PCR did not show a reliable correlation with viable plate counts or the colony hybridization assay. C. perfringens killed by irradiation were not detected by the plate count or colony hybridization methods; however, killed cells were detected with the PCR technique. By relying on the growth of viable cells for detection and/or enumeration, the colony hybridization and plate count methods provided a direct correlation with the presence of viable bacteria

Short communication: Lactose enhances bile tolerance of yogurt culture bacteria.

Science.gov (United States)

Mena, Behannis; Aryana, Kayanush

2018-03-01

Lactose is an energy source for culture bacteria. Bile tolerance is an important probiotic property. Our aim was to elucidate the effect of lactose on bile tolerance of yogurt starter culture Lactobacillus bulgaricus LB-12 and Streptococcus thermophilus ST-M5. Bile tolerance of pure cultures was determined using 0.3% oxgall in MRS THIO broth (Difco, Becton Dickinson, Sparks, MD) for L. bulgaricus and 0.3% oxgall in M17 broth (Oxoid, Basingstoke, UK) for Strep. thermophilus. Lactose was added to both broths at 0 (control), 1, 3, and 5% (wt/vol) broth. Dilutions were plated hourly for 12 h. Experiments were replicated 3 times. At 2, 4, and 12 h of incubation, lactose incorporated at all amounts, 1, 3, and 5% (wt/vol), showed higher counts of Strep. thermophilus ST-M5 compared with the control. Lactose use at 5% (wt/vol) significantly enhanced bile tolerance of both L. bulgaricus and Strep. thermophilus compared with control. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Efficacy of low-pressure foam cleaning compared to conventional cleaning methods in the removal of bacteria from surfaces associated with convenience food.

Science.gov (United States)

Lambrechts, A A; Human, I S; Doughari, J H; Lues, J F R

2014-09-01

Food borne illnesses and food poisoning are cause for concern globally. The diseases are often caused by food contamination with pathogenic bacteria due largely to poor sanitary habits or storage conditions. Prevalence of some bacteria on cleaned and sanitised food contact surfaces from eight convenience food plants in Gauteng (South Africa) was investigated with the view to evaluate the efficacy of the cleaning methods used with such food contact surfaces. The microbial load of eight convenience food manufacturing plants was determined by sampling stainless steel food contact surfaces after they had been cleaned and sanitised at the end of a day's shift. Samples were analysed for Total Plate Count (TPC), Escherichia coli, Salmonella species, Staphylococcus aureus and Listeria species. Results showed that 59 % of the total areas sampled for TPC failed to comply with the legal requirements for surfaces, according to the Foodstuffs, Cosmetics and Disinfectants Act ( 0.05) in terms of Listeria species isolates obtained from both cleaning methods. The LPF method proved to be the superior cleaning option for lowering TPC counts. Regardless of cleaning method used, pathogens continued to flourish on various surfaces, including dry stainless steel, posing a contamination hazard for a considerable period depending on the contamination level and type of pathogen. Intensive training for proper chemical usage and strict procedural compliance among workers for efficient cleaning procedures is recommended.

High precision refractometry based on Fresnel diffraction from phase plates.

Science.gov (United States)

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.

Total lactic acid bacteria, antioxidant activity, and acceptance of synbiotic yoghurt with red ginger extract (Zingiberofficinale var. rubrum)

Science.gov (United States)

Larasati, B. A.; Panunggal, B.; Afifah, D. N.; Anjani, G.; Rustanti, N.

2018-02-01

Antioxidant related to oxidative stress can caused the metabolic disorders. A functional food that high in antioxidant can be use as the alternative prevention. The addition of red ginger extract in yoghurt could form a functional food, that high in antioxidant, synbiotic and fiber. The influence of red ginger extract on yoghurt synbiotic against lactic acid bacteria, antioxidant activity and acceptance were analyzed. This was an experimental research with one factor complete randomized design, specifically the addition of red ginger extract 0%; 0,1%; 0,3% and 0,5% into synbiotic yoghurt. Total plate count method used to analyze the lactic acid bacteria, 1-1-diphenyl-2-picrylhydrazyl (DPPH) method for antioxidant activity, and acceptance analyzed with hedonic test. The higher the dose of extract added to synbiotic yoghurt, the antioxidant activity got significantly increased (ρ=0,0001), while the lactic acid bacteria got insignificantly decreased (ρ=0,085). The addition of 0,5% red ginger extract obtained the antioxidant activity of 71% and 4,86 × 1013 CFU/ml on lactic acid bacteria, which the requirement for probiotic on National Standard of Indonesia is >107 CFU/ml. The addition of extract had a significant effect on acceptance (ρ=0,0001) in flavor, color, and texture, but not aroma (ρ=0,266). The optimal product in this research was the yoghurt synbiotic with addition of 0,1% red ginger extract. To summarize, the addition of red ginger extract in synbiotic yoghurt had significant effect on antioxidant activity, flavor, color, and texture, but no significant effect on lactic acid bacteria and aroma.

Growth of Chlorella vulgaris and associated bacteria in photobioreactors

Science.gov (United States)

Lakaniemi, Aino‐Maija; Intihar, Veera M.; Tuovinen, Olli H.; Puhakka, Jaakko A.

2012-01-01

Summary The aim of this study was to test three flat plate photobioreactor configurations for growth of Chlorella vulgaris under non‐axenic conditions and to characterize and quantify associated bacterial communities. The photobioreactor cultivations were conducted using tap water‐based media to introduce background bacterial population. Growth of algae was monitored over time with three independent methods. Additionally, the quantity and quality of eukaryotes and bacteria were analysed using culture‐independent molecular tools based on denaturing gradient gel electrophoresis (PCR‐DGGE) and quantitative polymerase chain reaction (QPCR). Static mixers used in the flat plate photobioreactors did not generally enhance the growth at the low light intensities used. The maximum biomass concentration and maximum specific growth rate were 1.0 g l−1 and 2.0 day−1 respectively. Bacterial growth as determined by QPCR was associated with the growth of C. vulgaris. Based on PCR‐DGGE, bacteria in the cultures mainly originated from the tap water. Bacterial community profiles were diverse but reproducible in all flat plate cultures. Most prominent bacteria in the C. vulgaris cultures belonged to the class Alphaproteobacteria and especially to the genus Sphingomonas. Analysis of the diversity of non‐photosynthetic microorganisms in algal mass cultures can provide useful information on the public health aspects and unravel community interactions. PMID:21936882

Distribution of bacteria in frozen shrimps and their decontamination by gamma irradiation

Energy Technology Data Exchange (ETDEWEB)

Ito, Hitoshi; Adulyatham, P.; Ishigaki, Isao

1987-11-01

Six samples of frozen shrimps were obtained through importing company mainly from South-east Asian countries. Five samples of frozen shrimps contained 1.5 x 10/sup 5/ to 6.2 x 10/sup 6/ per gram of total aerobic bacteria, and 4 samples contained 1.4 x 10/sup 2/ to 2.2 x 10/sup 3/ per gram of coliforms, which are higher contamination than Japanese standard of hygenic level for frozen food products (total counts below 10/sup 5/ per gram and no coliforms). The dominant bacteria in total counts were consisted with psychrotrophic Moraxella, Arthrobacter, Micrococcus, Acinetobacter, Flavobacterium, Corynebacterium, Pseudomonas and etc, in all kinds of frozen shrimps. Salmonellae or enteric Vibrio were could not isolated by ordinary isolation methods, and there were isolated many strains of Proteus mirabilis instead of these bacteria. Necessary dose of gamma irradiation for decontamination of bacteria to hygenic level was decided as 3 approx. 5 kGy from the inactivation curves of total bacteria, coliforms, Proteus mirabilis and Salmonella typhymurium in frozen shrimps. Off-odor was suppressed by frozen condition of irradiation.

Distribution of bacteria in frozen shrimps and their decontamination by gamma irradiation

International Nuclear Information System (INIS)

Ito, Hitoshi; Adulyatham, P.; Ishigaki, Isao

1987-01-01

Six samples of frozen shrimps were obtained through importing company mainly from South-east Asian countries. Five samples of frozen shrimps contained 1.5 x 10 5 to 6.2 x 10 6 per gram of total aerobic bacteria, and 4 samples contained 1.4 x 10 2 to 2.2 x 10 3 per gram of coliforms, which are higher contamination than Japanese standard of hygenic level for frozen food products (total counts below 10 5 per gram and no coliforms). The dominant bacteria in total counts were consisted with psychrotrophic Moraxella, Arthrobacter, Micrococcus, Acinetobacter, Flavobacterium, Corynebacterium, Pseudomonas and etc, in all kinds of frozen shrimps. Salmonellae or enteric Vibrio were could not isolated by ordinary isolation methods, and there were isolated many strains of Proteus mirabilis instead of these bacteria. Necessary dose of gamma irradiation for decontamination of bacteria to hygenic level was decided as 3 ∼ 5 kGy from the inactivation curves of total bacteria, coliforms, Proteus mirabilis and Salmonella typhymurium in frozen shrimps. Off-odor was suppressed by frozen condition of irradiation. (author)

Sulfate-reducing bacteria colonize pouches formed for ulcerative colitis but not for familial adenomatous polyposis.

LENUS (Irish Health Repository)

Duffy, M

2012-02-03

PURPOSE: Ileal pouch-anal anastomosis remains the "gold standard" in surgical treatment of ulcerative colitis and familial adenomatous polyposis. Pouchitis occurs mainly in patients with a background of ulcerative colitis, although the reasons for this are unknown. The aim of this study was to characterize differences in pouch bacterial populations between ulcerative colitis and familial adenomatous pouches. METHODS: After ethical approval was obtained, fresh stool samples were collected from patients with ulcerative colitis pouches (n = 10), familial adenomatous polyposis (n = 7) pouches, and ulcerative colitis ileostomies (n = 8). Quantitative measurements of aerobic and anaerobic bacteria were performed. RESULTS: Sulfate-reducing bacteria were isolated from 80 percent (n = 8) of ulcerative colitis pouches. Sulfate-reducing bacteria were absent from familial adenomatous polyposis pouches and also from ulcerative colitis ileostomy effluent. Pouch Lactobacilli, Bifidobacterium, Bacteroides sp, and Clostridium perfringens counts were increased relative to ileostomy counts in patients with ulcerative colitis. Total pouch enterococci and coliform counts were also increased relative to ileostomy levels. There were no significant quantitative or qualitative differences between pouch types when these bacteria were evaluated. CONCLUSIONS: Sulfate-reducing bacteria are exclusive to patients with a background of ulcerative colitis. Not all ulcerative colitis pouches harbor sulfate-reducing bacteria because two ulcerative colitis pouches in this study were free of the latter. They are not present in familial adenomatous polyposis pouches or in ileostomy effluent collected from patients with ulcerative colitis. Total bacterial counts increase in ulcerative colitis pouches after stoma closure. Levels of Lactobacilli, Bifidobacterium, Bacteroides sp, Clostridium perfringens, enterococci, and coliforms were similar in both pouch groups. Because sulfate-reducing bacteria are

Effect of radiation decontamination on drug-resistant bacteria

International Nuclear Information System (INIS)

Ito, Hitoshi

2006-01-01

More than 80% of food poisoning bacteria such as Salmonella are reported as antibiotic-resistant to at least one type antibiotic, and more than 50% as resistant to two or more. For the decontamination of food poisoning bacteria in foods, radiation resistibility on drug-resistant bacteria were investigated compared with drug-sensitive bacteria. Possibility on induction of drug-resistant mutation by radiation treatment was also investigated. For these studies, type strains of Escherichia coli S2, Salmonella enteritidis YK-2 and Staphylococcus aureus H12 were used to induce drug-resistant strains with penicillin G. From the study of radiation sensitivity on the drug-resistant strain induced from E. coli S2, D 10 value was obtained to be 0.20 kGy compared with 0.25 kGy at parent strain. On S. enteritidis YK-2, D 10 value was obtained to be 0.14 kGy at drug-resistant strain compared with 0.16 kGy at parent strain. D 10 value was also obtained to be 0.15 kGy at drug-resistant strain compared with 0.21 kGy at parent strain of St. aureus H12. Many isolates of E. coli 157:H7 or other type of E. coli from meats such as beef were resistant to penicillin G, and looked to be no relationship on radiation resistivities between drug-resistant strains and sensitive strains. On the study of radiation sensitivity on E. coli S2 at plate agars containing antibiotics, higher survival fractions were obtained at higher doses compared with normal plate agar. The reason of higher survival fractions at higher doses on plate agar containing antibiotics should be recovery of high rate of injured cells by the relay of cell division, and drug-resistant strains by mutation are hardly induced by irradiation. (author)

Use of Gamma Rays to Control of Internal Parasites and Pathogenic Bacteria in Silver side Fish (Bissaria)

International Nuclear Information System (INIS)

Shawki, H.A.; El-Hanafy, A.E.A.; Shagar, G.I.A.

2014-01-01

The usefulness of Gamma irradiation to control of internal parasites and pathogenic bacteria found in silver side fish (Atherina) (Bissaria) was investigated. The detected parasites and the prevalence were adult Trematode (37.5%); Cestodes (95%) and Nematode (22.5%) for control silver side fish. The counts of detected microorganisms, (Total bacterial count; Psychrophilic bacteria; Mold and yeast; E.coli and Staphyloccous aureus ) were 4.89 ; 2.30; 2.32; 2.31 and 2.04 log 10 cfu/g for control silver side fish, respectively. Applied irradiation doses reduce the infected rate by Trematode, Cestodes and Nematode, furthermore, gamma irradiation with different doses (0.5, 1, 2, 4 and 5 kGy) reduce the microorganisms count of silver side fish (Bissaria) samples and the rate of decrement increase with the dose increase. Total bacterial count was not detected by using dose 5 kGy while Psychrophilic bacteria were completely eliminated using dose 1 kGy. On the other hand mold and yeast; E.coli and Staph aureus in silver side fish samples were not detected after subjected to gamma irradiation with dose 4 kGy. The results suggest that the applied doses completely elimination different internal parasites and pathogenic bacteria found in silver side fish. Thus, it could be conclude that the irradiation dose of 4 kGy can be effectively applied to ensure the safety of internal parasites and pathogenic bacteria found in silver side fish (Atherina) (Bissaria) with regards to these harmful parasites and pathogenic bacteria

Cadmium resistance of endophytic bacteria and rizosféricas bacteria isolated from Oriza sativa in Colombia

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Nataly Ayubb T

2017-12-01

Full Text Available The present study had as objective to evaluate in vitro the resistance of endophytic bacteria and rizospheric bacteria to different concentrations of Cadmium.This bacteria were isolated fron different tissues of commercial rice varieties and from bacteria isolated from the rhizosphere in rice plantations of the Nechí (Antioquía and Achí (Bolivar.  Plant growth promotion was evaluated in vitro by nitrogen fixation, phosphate solubilization and siderophores production of endophytic bacteria. Of each tissue isolated from rice plants was carried out isolation in culture medium for endophytic bacteria, and the soil samples were serially diluted in peptone water. Each sample was determined the population density by counting in CFU / g of tissue and morphotypes were separated by shape, color, size and appearance in culture media. Significant differences were observed for density population of bacteria with respect to tissue, with higher values in root (4x1011 g/root, followed of the stem (3x1010g/etem, leaf (5x109 g/ leaf, flag leaf (3x109 g/ flag leaf and with less density in panicle (4x108 g/panicle. The results of the identification with kit API were confirmed the presence of endophytic bacteria Burkholderia cepaceae and rizospheric bacteria Pseudomona fluorescens With the ability to tolerate different concentrations of Cd, fix nitrogen, solubilize phosphates and produce siderophores.

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

Science.gov (United States)

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.

Simplified quality control of radioactive seeds in sterile cartridge for prostate brachytherapy using an imaging plate and a collimator

International Nuclear Information System (INIS)

Saze, T.; Miyoshi, H.; Maezawa, H.; Kubota, M.; Furutani, S.; Nishitani, H.; Kawaguchi, Y.; Nakayama, S.; Ito, S.; Nishizawa, K.

2008-01-01

Image analyzer system and collimator has been successfully applied to calibrate simultaneously multiple 125 I seeds in a sterile cartridge. Seeds within the cartridge were placed on an imaging plate, and the imaging plate irradiated. To remove scatter radiation, and improve spatial resolution of seed images, this study used a specially designed collimator. The irradiated imaging plate was scanned using an image analyzer system, and radioactivity intensities of seed images were given in counts. Counts could be translated to profiles, and each seed within the cartridge was analyzed. It is observed that a good correlation between counts and total radioactivity of the seeds within the cartridge. Thus, using a least-squares line, it was possible to calibrate a seed with unknown apparent activity. By analyzing the profiles, it was possible not only to detect a mis calibrated seed in the cartridge from its relative difference in counts, but also to identify its position in the cartridge. Using an imaging analyzer system, all seeds in a cartridge could be calibrated in a sterile environment. (author)

The inhibitory activity of Lactic acid bacteria isolated from fresh cow cheese

Directory of Open Access Journals (Sweden)

Nevijo Zdolec

2007-04-01

Full Text Available Lactic acid bacteria are the constituent part of milk microbial flora that could influence the safety of dairy products due production of organic acids, hydrogen peroxide, carbon dioxide and bacteriocins. Taking this in consideration, the objective of this study was to investigate the composition of lactic acid bacteria population in fresh cow cheeses taken from local markets, as well as their antimicrobial capacity. Lactic acid bacteria counts were determined according to ISO 1524:1998 method, biochemical determination using API 50 CHL system, and inhibitory activity against L. monocytogenes NCTC 10527 by agar well diffusion assay. Lactic acid bacteria count in fresh cow cheeses (n=10 ranged from 5.87 to 8.38 log10 CFU g-1. Among 52 MRS isolates collected, 61.54 % were assigned to the Lactococcus lactis subsp. Lactis species, 23.07 % Lactobacillus helveticus, 11.54 % Leuconostoc mesenteroides subsp. cremoris and 3.85 % Leuconostoc mesenteroides subsp. mesenteroides. Antilisterial activity was found in 18 isolates.

Fungi outcompete bacteria under increased uranium concentration in culture media

International Nuclear Information System (INIS)

Mumtaz, Saqib; Streten-Joyce, Claire; Parry, David L.; McGuinness, Keith A.; Lu, Ping; Gibb, Karen S.

2013-01-01

As a key part of water management at the Ranger Uranium Mine (Northern Territory, Australia), stockpile (ore and waste) runoff water was applied to natural woodland on the mine lease in accordance with regulatory requirements. Consequently, the soil in these Land Application Areas (LAAs) presents a range of uranium concentrations. Soil samples were collected from LAAs with different concentrations of uranium and extracts were plated onto LB media containing no (0 ppm), low (3 ppm), medium (250 ppm), high (600 ppm) and very high (1500 ppm) uranium concentrations. These concentrations were similar to the range of measured uranium concentrations in the LAAs soils. Bacteria grew on all plates except for the very high uranium concentrations, where only fungi were recovered. Identifications based on bacterial 16S rRNA sequence analysis showed that the dominant cultivable bacteria belonged to the genus Bacillus. Members of the genera Paenibacillus, Lysinibacillus, Klebsiella, Microbacterium and Chryseobacterium were also isolated from the LAAs soil samples. Fungi were identified by sequence analysis of the intergenic spacer region, and members of the genera Aspergillus, Cryptococcus, Penicillium and Curvularia were dominant on plates with very high uranium concentrations. Members of the Paecilomyces and Alternaria were also present but in lower numbers. These findings indicate that fungi can tolerate very high concentrations of uranium and are more resistant than bacteria. Bacteria and fungi isolated at the Ranger LAAs from soils with high concentrations of uranium may have uranium binding capability and hence the potential for uranium bioremediation. -- Highlights: ► Fungi outcompete bacteria under increased uranium concentration in culture media. ► Soil microorganisms isolated from the Ranger Land Application Areas (LAAs) were resistant to uranium. ► Bacillus was the most abundant cultivable genus retrieved from the Ranger LAAs soils. ► Uranium in LAAs soils is

Characterization of anti-listerial lactic acid bacteria isolated from Thai fermented fish products

DEFF Research Database (Denmark)

Østergaard, Anya; Embarek, Peter Karim Ben; Wedell-Neergaard, C.

1998-01-01

Thai fermented fish products were screened for lactic acid bacteria capable of inhibiting Listeria sp. (Listeria innocua). Of 4150 assumed lactic acid bacteria colonies from MRS agar plates that were screened by an agar-overlay method 58 (1.4%) were positive. Forty four of these strains were...

Effects of Ethanolic Ferolagu angulata Extract on Pathogenic Gastrointestinal Bacteria and Probiotic Bacteria in Skimmed Milk Medium

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Reza Naghiha

2016-12-01

Full Text Available Background:    Due to excessive consumption of synthetic drugs, drug resistance rate of pathogenic bacteria is increasing and there is an ever-increasing need to find new safe compounds to tackle this problem. This study was conducted to investigate the consequences of chavill extract on the growth and viability of gastrointestinal pathogenic bacterium and probiotics bacteria. Methods:    The experiment contained three levels of the chavill extract concentrations (0, 1 and 3% which were added to the milk free fat in accompany with three probiotic bacteria (Lactobacillus acidophilus, Lactobacillus casei and lactobacillus plantaram and a pathogenic gastrointestinal bacterium (Salmonella typhimurium. Bacterial inoculums (1×107 CFU/ml with different concentrations of chavill extract were added to skimmed milk medium and bacteria growth were enumerated. Results:  The concentration of 1% chavill extract significantly increased the total count of probiotic bacteria compared to the control group, while the number of pathogenic bacteria was decreased. At 3% chavill extract the growth of Lactobacillus acidophilus and Lactobacillus plantaram were increased. On the other hand, it prevented the growth of Salmonella typhimurium Conclusion:   Chavill extracts would play as an alternative to antibiotics in pharmacological studies to decreases harmful bacteria and increase probiotic bacteria.

Micro-plate radiobinding assay of autoantibody to glutamic acid decarboxylase

International Nuclear Information System (INIS)

Huang Gan; Jin Helai; Wang Xia; Li Hui; Zhang Song; Zhou Zhiguang

2008-01-01

Objective: The purpose of this study was to develop a high-throughput micro-plate radiobinding assay (RBA) of glutamic acid decarboxylase antibody (CAD-Ab) and to evaluate its clinical application. Methods: 35 S labeled GAD 65 antigen was incubated with sera for 24 h on a 96-well plate, and then transferred to the Millipore plate coated with protein A, which was washed with 4 degree C PBS buffer, and then counted by a liquid scintillation counter. The CAD-Ab results were expressed by WHO standard unit (U/ml). A total of 224 healthy controls, 162 patients with type 1 diabetes mellitus (T1DM) and 210 patients with newly diagnosed type 2 diabetes (T2DM) were recruited. A total of 119 T1DM and healthy eases with gradually changing GAD-Ab levels were selected to compare the consistency of micro-plate RBA with conventional radioligand assay (RLA). Blood samples were obtained from the peripheral vein and finger tip in 32 healthy controls, 35 T1DM and 24 T2DM patients, and tested with micro-plate RBA and then compared with the conventional RLA to investigate the reliability of finger tip sampling. Linear correlation, student's t-test, variance analysis and receiver operating characteristic (ROC) curve were performed using SPSS 11.5. Results: (1) The optimized conditions of micro-plate RBA included 2 μl serum incubated with 3 x 10 4 counts/min 35 S-CAD for 24 h under slow vibration, antigen-antibody compounds washed 10 times by 4 degree C PBS buffer, and radioactivity counted with Optiphase Supermix scintillation liquid. (2)The intra-batch CV of the micro-plate RBA was 3.8%-10.2%, and the inter-batch CV was 5.6%-11.9%. The linearity analysis showed a good correlation when the GAD-Ab in serum samples ranged from 40.3 to 664 U/ml and the detection limit of measurement was 3.6 U/ml. The results from Diabetes Autoantibody Standardization Program (DASP) 2005 showed that the sensitivity and specificity for GAD-Ab were 78% (39 positive among 50 new-onset T1DM) and 98% (2 positive

Metal oxide/hydroxide-coated dual-media filter for simultaneous removal of bacteria and heavy metals from natural waters.

Science.gov (United States)

Ahammed, M Mansoor; Meera, V

2010-09-15

The present study was conducted to compare the performance of a dual-media filter consisting of manganese oxide-coated (MOCS) and iron hydroxide-coated sand (IOCS) with that of IOCS filter and uncoated sand filter in treating water contaminated by microorganisms, heavy metals and turbidity with a view to its use in simple household water purification devices in developing countries. Long-duration column tests were conducted using two natural waters namely, roof-harvested rainwater and canal water. Performance of the filters showed that dual-media filter was more efficient in removing bacteria and heavy metals compared to IOCS filter, while uncoated sand filter showed very poor performance. The average effluent levels for dual-media filter when tested with rainwater were: turbidity 1.0+/-0.1 NTU; total coliforms 3+/-2 MPN/100 mL; heterotrophic plate count 170+/-20 CFU/mL; zinc 0.06+/-0.01 mg/L, while that for IOCS filter were: turbidity 1.0+/-0.1 NTU; total coliforms 4+/-2 MPN/100 mL; heterotrophic plate count 181+/-37 CFU/mL; zinc 0.20+/-0.07 mg/L. Similar results were obtained for canal water also. Up to 900 bed volumes (BV) could be treated without affecting the efficiency in the case of rainwater, while the filter operation had to be terminated after 500 BV due to excessive headloss in the case of canal water. The study thus showed the potential of the dual-media for use in low-cost household water filters for purification of natural waters. Copyright 2010 Elsevier B.V. All rights reserved.

A preliminary study of anaerobic thiosulfate-oxidising bacteria as denitrifiers in the Arabian Sea

Digital Repository Service at National Institute of Oceanography (India)

LokaBharathi, P.A.; Chandramohan, D.; Nair, S.

Bacteria which oxidize thiosulfate and reduce nitrate (TONRB) and bacteria which oxidize thiosulfate and denitrify (TODB) sampled at 5-, 100-, 200-and 300-m depths were enumerated in agar shake cultures by colony counting and by applying MPN...

Single electron counting using a dual MCP assembly

International Nuclear Information System (INIS)

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

2016-01-01

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. - Highlights: • Dual MCPs assembly with four electrodes using different voltage combinations has been investigated for single electron counting. • Both the MCP voltages and the gap voltage can affect the gain, pulse height resolution and P/V ratio. • A high gain of the first stage MCP, a saturation mode of the second stage MCP and an appropriately reverse gap voltage can improve the resolution greatly. • The optimum voltage arrangements is significant for the design of MCP detectors in single electron counting applications.

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

Science.gov (United States)

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.

Influence of flooring type during transport and holding on bacteria recovery from broiler carcass rinses before and after defeathering.

Science.gov (United States)

Buhr, R J; Cason, J A; Dickens, J A; Hinton, A; Ingram, K D

2000-03-01

Four trials were conducted to determine whether conventional solid or elevated wire mesh flooring, during transport and holding of broilers prior to slaughter, influenced the number of bacteria recovered from feathered and defeathered carcasses. After 4 h off feed, 7-wk-old broilers were placed at commercial density into a modified commercial transport dump-coop on either fiberglass sheeting or 2.54x2.54 cm wire mesh flooring that allowed feces to fall through. Broilers were transported for 1 h and then held for 13 h under a covered shed before processing. Broilers were killed by electrocution, and the vents were plugged to prevent escape of feces. External carcass rinses were obtained twice (from the same carcass) from eight broilers per flooring treatment per trial, before scalding and defeathering and again after defeathering and removal of the head and feet. Greater numbers of total aerobes, coliforms, and Escherichia coli were recovered from feathered carcasses than from defeathered carcasses. Campylobacter count was also less for defeathered than feathered carcasses from the solid flooring treatment but did not significantly decrease following defeathering of carcasses from the wire flooring. The incidence of Campylobacter-positive carcasses was reduced following defeathering for both flooring treatments, but the percentage of Salmonellae-positive carcasses remained constant. Coliform (log10 6.20 vs. 5.63 cfu/mL of rinse) and E. coli (log10 5.93 vs. 5.36) counts in the feathered rinses were significantly higher for the solid flooring compared with wire flooring, respectively. After defeathering, the number of coliforms (log10 3.12) and E. coli (log10 2.91) recovered did not differ between flooring treatments. Aerobic plate count (log10 7.06 and 4.02), Campylobacter count (log10 2.49 and 1.80), and the incidence of Campylobacter-positive (44 and 11%) and Salmonellae-positive (52 and 50%) carcasses for feathered and defeathered rinses, respectively, did not

Modulation of cell surface hydrophobicity and attachment of bacteria to abiotic surfaces and shrimp by Malaysian herb extracts.

Science.gov (United States)

Hui, Yew Woh; Dykes, Gary A

2012-08-01

The use of simple crude water extracts of common herbs to reduce bacterial attachment may be a cost-effective way to control bacterial foodborne pathogens, particularly in developing countries. The ability of water extracts of three common Malaysian herbs (Andrographis paniculata, Eurycoma longifolia, and Garcinia atroviridis) to modulate hydrophobicity and attachment to surfaces of five food-related bacterial strains (Bacillus cereus ATCC 14576, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 10145, Salmonella Enteritidis ATCC 13076, Staphylococcus aureus ATCC 25923) were determined. The bacterial attachment to hydrocarbon assay was used to determine bacterial hydrophobicity. Staining and direct microscopic counts were used to determine attachment of bacteria to glass and stainless steel. Plating on selective media was used to determine attachment of bacteria to shrimp. All extracts were capable of either significantly ( P 0.05) to bacterial attachment. For specific combinations of bacteria, surface material, and plant extract, significant correlations (R > 0.80) between hydrophobicity and attachment were observed. The highest of these was observed for S. aureus attachment to stainless steel and glass after treatment with the E. longifolia extract (R = 0.99, P < 0.01). The crude water herb extracts in this study were shown to have the potential to modulate specific bacterial and surface interactions and may, with further work, be useful for the simple and practical control of foodborne pathogens.

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

Science.gov (United States)

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

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

Directory of Open Access Journals (Sweden)

Muhammed Majeed

Full Text Available 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.

Science.gov (United States)

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.

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

Science.gov (United States)

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

Counting mycobacteria in infected human cells and mouse tissue: a comparison between qPCR and CFU.

Directory of Open Access Journals (Sweden)

Sharad Pathak

Full Text Available Due to the slow growth rate and pathogenicity of mycobacteria, enumeration by traditional reference methods like colony counting is notoriously time-consuming, inconvenient and biohazardous. Thus, novel methods that rapidly and reliably quantify mycobacteria are warranted in experimental models to facilitate basic research, development of vaccines and anti-mycobacterial drugs. In this study we have developed quantitative polymerase chain reaction (qPCR assays for simultaneous quantification of mycobacterial and host DNA in infected human macrophage cultures and in mouse tissues. The qPCR method cannot discriminate live from dead bacteria and found a 10- to 100-fold excess of mycobacterial genomes, relative to colony formation. However, good linear correlations were observed between viable colony counts and qPCR results from infected macrophage cultures (Pearson correlation coefficient [r] for M. tuberculosis = 0.82; M. a. avium = 0.95; M. a. paratuberculosis = 0.91. Regression models that predict colony counts from qPCR data in infected macrophages were validated empirically and showed a high degree of agreement with observed counts. Similar correlation results were also obtained in liver and spleen homogenates of M. a. avium infected mice, although the correlations were distinct for the early phase (< day 9 post-infection and later phase (≥ day 20 post-infection liver r = 0.94 and r = 0.91; spleen r = 0.91 and r = 0.87, respectively. Interestingly, in the mouse model the number of live bacteria as determined by colony counts constituted a much higher proportion of the total genomic qPCR count in the early phase (geometric mean ratio of 0.37 and 0.34 in spleen and liver, respectively, as compared to later phase of infection (geometric mean ratio of 0.01 in both spleen and liver. Overall, qPCR methods offer advantages in biosafety, time-saving, assay range and reproducibility compared to colony counting. Additionally, the duplex format allows

Cellular viability of Saccharomyces cerevisiae cultivated in association with contaminates bacteria of alcoholic fermentation

International Nuclear Information System (INIS)

Nobre, Thais de Paula

2005-01-01

The aim of this work was to study the influence of the bacteria Bacillus and Lactobacillus, as well as their metabolic products, in reduction of cellular viability of Saccharomyces cerevisiae, when in mixed culture of yeast and active and treated bacteria. Also was to evaluated an alternative medium (MCC) for the cultivation of bacteria and yeast, constituted of sugarcane juice diluted to 5 deg Brix and supplemented with yeast extract and peptone. The bacteria Bacillus subtilis, Bacillus coagulans, Bacillus stearothermophilus, Lactobacillus fermentum and Lactobacillus plantarum were cultivated in association with yeast Saccharomyces cerevisiae (strain Y-904) for 72 h on 32 deg C, under agitation. The cellular viability, budding rate and population of S. cerevisiae, the total acidity, volatile acidity and pH of culture were determined from 0, 24, 48 e 72 h of mixed culture. Also were determined the initial and final of microorganism population across the pour plate method, in traditional culture medium (PCA for Bacillus, MRS-agar for Lactobacillus and YEPD-agar for yeast S. cerevisiae) and in medium constituted of sugarcane juice. The bacteria cultures were treated by heat sterilization (120 deg C for 20 minutes), antibacterial agent (Kamoran HJ in concentration 3,0 ppm) or irradiation (radiation gamma, with doses of 5,0 kGy for Lactobacillus and 15,0 kGy for Bacillus). The results of the present research showed that just the culture mediums more acids (with higher concentrations of total and volatile acidity, and smaller values of pH), contaminated with active bacteria L. fermentum and B. subtilis, caused reduction on yeast cellular viability. Except the bacteria B. subtilis treated with radiation, the others bacteria treated by different procedures (heat, radiation e antibacterial) did not cause reduction on yeast cellular viability and population, indicating that the isolated presence of the cellular metabolic of theses bacteria was not enough to reduce the

High hydrostatic pressure inactivation of total aerobic bacteria, lactic acid bacteria, yeasts in sour Chinese cabbage.

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Li, Lin; Feng, Lun; Yi, Junjie; Hua, Cheng; Chen, Fang; Liao, Xiaojun; Wang, Zhengfu; Hu, Xiaosong

2010-08-15

This study investigated the inactivation of total aerobic bacteria (TAB), lactic acid bacteria (LAB), yeasts in sour Chinese cabbage (SCC) treated by high hydrostatic pressure (HHP). The pressure level ranged from 200 to 600 MPa and the treatment time were 10-30 min. All samples were stored at 4, 27 and 37 degrees C for 90 days. The pressure level of 200 MPa had no significant impact on these microorganisms. The counts of TAB were significantly reduced by 2.7-4.0 log(10)CFU/g at 400 MPa and 4.2-4.5 log(10)CFU/g at 600 MPa from 6.2 log(10)CFU/g; the counts of LAB were also reduced by 2.4-4.3 log(10)CFU/g at 400 MPa from 7.0 log(10)CFU/g and LAB was completely inactivated at 600 MPa; the counts of yeasts were reduced by 1.5-2.0 log(10)CFU/g at 400 and 600 MPa from 4.2 log(10)CFU/g. Storage temperatures significantly influenced the microbial proliferation in HHP-treated SCC depending on the pressure levels. The surviving TAB and LAB at 400 MPa equaled initial counts after 15-day storage at 27 and 37 degrees C, whereas they were inhibited at 4 degrees C up to 60 days. The surviving TAB at 600 MPa did not grow. Yeasts at 400 and 600 MPa decreased below detectable level after 2 days at all the three storage temperatures. From the microbial safety point of view, the result indicated that HHP at 600 MPa could be used as an alternative preservation method for SCC. Copyright 2010 Elsevier B.V. All rights reserved.

Development and characterisation of a visible light photon counting imaging detector system

CERN Document Server

Barnstedt, J

2002-01-01

We report on the development of a visible light photon counting imaging detector system. The detector concept is based on standard 25 mm diameter microchannel plate image intensifiers made by Proxitronic in Bensheim (Germany). Modifications applied to these image intensifiers are the use of three microchannel plates instead of two and a high resistance ceramics plate used instead of the standard phosphor output screen. A wedge and strip anode mounted directly behind the high resistance ceramics plate was used as a read out device. This wedge and strip anode picks up the image charge of electron clouds emerging from the microchannel plates. The charge pulses are fed into four charge amplifiers and subsequently into a digital position decoding electronics, achieving a position resolution of up to 1024x1024 pixels. Mounting the anode outside the detector tube is a new approach and has the great advantage of avoiding electrical feedthroughs from the anode so that the standard image intensifier fabrication process...

Effects of gamma-rays on an indigenous Bacillus isolate

International Nuclear Information System (INIS)

Jong Bor Chyan; Liew Pauline Woan Ying; Ahmad Zainuri Mohd Dzomir; Siti Khadijah Abu Hadin; Nabilahuda Mohd Tumirin

2010-01-01

A preliminary study was carried out with the aim to obtain strong cellulolytic bacteria by ionizing radiation using a 60 Co source. An indigenous cellulolytic Bacillus sp. NMBCC 10023 originally isolated from soil origin was used in the study. The harvested bacterial pellets from overnight growth cultures were exposed to gamma-irradiation with doses ranging from 1 kGy to 40 kGy. The numbers of surviving bacteria on agar plate decreased as the gamma irradiation dose increased. No isolates were recovered after exposure to doses greater than 10 kGy. Based on the cell count using plate count method, the 90 % lethal dose (LD90) of gamma radiation of Bacillus sp. NMBCC 10023 was between 2-4 kGy. (author)

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

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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.

Temperature Effect Study on Growth and Survival of Pathogenic Vibrio parahaemolyticus in Jinjiang Oyster (Crassostrea rivularis with Rapid Count Method

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Yuan Wang

2018-01-01

Full Text Available The growth of Vibrio parahaemolyticus (V. parahaemolyticus in oysters during postharvest storage increases the possibility of its infection in humans. In this work, to investigate the growth or survival profiles in different media, pathogenic V. parahaemolyticus in APW, Jinjiang oyster (JO, Crassostrea rivularis slurry, and live JO were studied under different temperatures. All the strain populations were counted through our double-layer agar plate (DLAP method. In APW, the pathogenic V. parahaemolyticus showed continuous growth under 15, 25, and 35°C, while a decline in behavior was displayed under 5°C. The similar survival trend of pathogenic V. parahaemolyticus in JO slurry and live JO was observed under 5, 25, and 35°C, except the delayed growth or decline profile compared to APW. Under 15°C, they displayed decline and growth profile in JO slurry and live JO, respectively. These results indicate the different sensitivity of pathogenic V. parahaemolyticus in these matrices to temperature variation. Furthermore, nonpathogenic V. parahaemolyticus displayed little difference in survival profiles when inoculated in live JO under corresponding temperatures. The results indicate that inhibition or promotion effect could be regulated under different storage temperature for both pathogenic and nonpathogenic strains. Besides, the DLAP method showed the obvious quickness and efficiency during the bacteria count.

Gravity separation of fat, somatic cells, and bacteria in raw and pasteurized milks.

Science.gov (United States)

Caplan, Z; Melilli, C; Barbano, D M

2013-04-01

The objective of experiment 1 was to determine if the extent of gravity separation of milk fat, bacteria, and somatic cells is influenced by the time and temperature of gravity separation or the level of contaminating bacteria present in the raw milk. The objective of experiment 2 was to determine if different temperatures of milk heat treatment affected the gravity separation of milk fat, bacteria, and somatic cells. In raw milk, fat, bacteria, and somatic cells rose to the top of columns during gravity separation. About 50 to 80% of the fat and bacteria were present in the top 8% of the milk after gravity separation of raw milk. Gravity separation for 7h at 12°C or for 22h at 4°C produced equivalent separation of fat, bacteria, and somatic cells. The completeness of gravity separation of fat was influenced by the level of bacteria in the milk before separation. Milk with a high bacterial count had less (about 50 to 55%) gravity separation of fat than milk with low bacteria count (about 80%) in 22h at 4°C. Gravity separation caused fat, bacteria, and somatic cells to rise to the top of columns for raw whole milk and high temperature, short-time pasteurized (72.6°C, 25s) whole milk. Pasteurization at ≥76.9°C for 25s prevented all 3 components from rising, possibly due to denaturation of native bovine immunoglobulins that normally associate with fat, bacteria, and somatic cells during gravity separation. Gravity separation can be used to produce reduced-fat milk with decreased bacterial and somatic cell counts, and may be a critical factor in the history of safe and unique traditional Italian hard cheeses produced from gravity-separated raw milk. A better understanding of the mechanism of this natural process could lead to the development of new nonthermal thermal technology (that does not involve heating the milk to high temperatures) to remove bacteria and spores from milk or other liquids. Copyright © 2013 American Dairy Science Association. Published by

Cell viability of mycorrhiza helper bacteria solid inoculant in different carrier material

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Asyiah, Iis Nur; Hindersah, Reginawanti; Harni, Rita

2018-02-01

Roots of food crops are colonized by nonpathogenic mycorrhizal fungi which show natural ability to control plant pathogen. Mycorrhizal establishment in plant roots is affected by rhizobacteria, known as mycorrhiza helper bacteria (MHB), which has synergetic effects on mycorrhizal associations. Laboratory experiment has been conducted to assess the best carrier material to develop well-qualified MHB of Pseudomonas diminuta and Bacillus subtilis solid inoculant. Carrier materials were 100 mesh organic matter of agricultural waste. Different spore concentration of both bacterial liquid inoculants were grown on three kinds of 100-mesh organic matter and stored at room temperature up to 90 days. Cell viability of both MHB were counted by serial dilution plate method by using specific medium. The results showed that sugar cane baggase ash was the best carrier material to maintain cell viability for both MHB. However, the population of Pseudomonas diminuta and Bacillus subtilis in sugar cane baggase ash were slightly decreased after 90 days. The use of sugarcane baggase ash for solid MHB inoculant development could be suggested.

Viability of sublethally injured coliform bacteria on fresh-cut cabbage stored in high CO2 atmospheres following rinsing with electrolyzed water.

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Izumi, Hidemi; Inoue, Ayano

2018-02-02

The extent of sublethally injured coliform bacteria on shredded cabbage, either rinsed or not rinsed with electrolyzed water, was evaluated during storage in air and high CO 2 controlled atmospheres (5%, 10%, and 15%) at 5°C and 10°C using the thin agar layer (TAL) method. Sublethally injured coliform bacteria on nonrinsed shredded cabbage were either absent or they were injured at a 64-65% level when present. Rinsing of shredded cabbage with electrolyzed water containing 25ppm available chlorine reduced the coliform counts by 0.4 to 1.1 log and caused sublethal injury ranging from 42 to 77%. Pantoea ananatis was one of the species injured by chlorine stress. When shredded cabbage, nonrinsed or rinsed with electrolyzed water, was stored in air and high CO 2 atmospheres at 5°C for 7days and 10°C for 5days, coliform counts on TAL plates increased from 3.3-4.5 to 6.5-9.0 log CFU/g during storage, with the increase being greater at 10°C than at 5°C. High CO 2 of 10% and 15% reduced the bacterial growth on shredded cabbage during storage at 5°C. Although injured coliform bacteria were not found on nonrinsed shredded cabbage on the initial day, injured coliforms at a range of 49-84% were detected on samples stored in air and high CO 2 atmospheres at 5°C and 10°C. Injured cells were detected more frequently during storage at both temperatures irrespective of the CO 2 atmosphere when shredded cabbage was rinsed with electrolyzed water. These results indicated that injured coliform bacteria on shredded cabbage, either rinsed or not rinsed with electrolyzed water, exhibited different degrees of injury during storage regardless of the CO 2 atmosphere and temperature tested. Copyright © 2017 Elsevier B.V. All rights reserved.

Effects of levan-type fructan on growth performance, nutrient digestibility, diarrhoea scores, faecal shedding of total lactic acid bacteria and coliform bacteria, and faecal gas emission in weaning pigs.

Science.gov (United States)

Lei, Xin Jian; Kim, Yong Min; Park, Jae Hong; Baek, Dong Heon; Nyachoti, Charles Martin; Kim, In Ho

2018-03-01

The use of antibiotics as growth promoters in feed has been fully or partially banned in several countries. The objective of this study was to evaluate effects of levan-type fructan on growth performance, nutrient digestibility, faecal shedding of lactic acid bacteria and coliform bacteria, diarrhoea scores, and faecal gas emission in weaning pigs. A total of 144 weaning pigs [(Yorkshire × Landrace) × Duroc] were randomly allocated to four diets: corn-soybean meal-based diets supplemented with 0, 0.1, 0.5, or 1.0 g kg -1 levan-type fructan during this 42-day experiment. During days 0 to 21 and 0 to 42, average daily gain and average daily feed intake were linearly increased (P bacteria counts were linearly increased (P = 0.001). The results indicate that dietary supplementation with increasing levan-type fructan enhanced growth performance, improved nutrient digestibility, and increased faecal lactic acid bacteria counts in weaning pigs linearly. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

A Repeating Sulfated Galactan Motif Resuscitates Dormant Micrococcus luteus Bacteria.

Science.gov (United States)

Böttcher, Thomas; Szamosvári, Dávid; Clardy, Jon

2018-07-01

Only a small fraction of bacteria can autonomously initiate growth on agar plates. Nongrowing bacteria typically enter a metabolically inactive dormant state and require specific chemical trigger factors or signals to exit this state and to resume growth. Micrococcus luteus has become a model organism for this important yet poorly understood phenomenon. Only a few resuscitation signals have been described to date, and all of them are produced endogenously by bacterial species. We report the discovery of a novel type of resuscitation signal that allows M. luteus to grow on agar but not agarose plates. Fractionation of the agar polysaccharide complex and sulfation of agarose allowed us to identify the signal as highly sulfated saccharides found in agar or carrageenans. Purification of hydrolyzed κ-carrageenan ultimately led to the identification of the signal as a small fragment of a large linear polysaccharide, i.e., an oligosaccharide of five or more sugars with a repeating disaccharide motif containing d-galactose-4-sulfate (G4S) 1,4-linked to 3,6-anhydro-α-d-galactose (DA), G4S-(DA-G4S) n ≥2 IMPORTANCE Most environmental bacteria cannot initiate growth on agar plates, but they can flourish on the same plates once growth is initiated. While there are a number of names for and manifestations of this phenomenon, the underlying cause appears to be the requirement for a molecular signal indicating safe growing conditions. Micrococcus luteus has become a model organism for studying this growth initiation process, often called resuscitation, because of its apparent connection with the persistent or dormant form of Mycobacterium tuberculosis , an important human pathogen. In this report, we identify a highly sulfated saccharide from agar or carrageenans that robustly resuscitates dormant M. luteus on agarose plates. We identified and characterized the signal as a small repeating disaccharide motif. Our results indicate that signals inherent in or absent from the

Quantification of viable spray-dried potential probiotic lactobacilli using real-time PCR

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Radulović Zorica

2012-01-01

Full Text Available The basic requirement for probiotic bacteria to be able to perform expected positive effects is to be alive. Therefore, appropriate quantification methods are crucial. Bacterial quantification based on nucleic acid detection is increasingly used. Spray-drying (SD is one of the possibilities to improve the survival of probiotic bacteria against negative environmental effects. The aim of this study was to investigate the survival of spray-dried Lactobacillus plantarum 564 and Lactobacillus paracasei Z-8, and to investigate the impact on some probiotic properties caused by SD of both tested strains. Besides the plate count technique, the aim was to examine the possibility of using propidium monoazide (PMA in combination with real-time polymerase chain reaction (PCR for determining spray-dried tested strains. The number of intact cells, Lb. plantarum 564 and Lb. paracasei Z-8, was determined by real-time PCR with PMA, and it was similar to the number of investigated strains obtained by the plate count method. Spray-dried Lb. plantarum 564 and Lb. paracasei Z-8 demonstrated very good probiotic ability. It may be concluded that the PMA real-time PCR determination of the viability of probiotic bacteria could complement the plate count method and SD may be a cost-effective way to produce large quantities of some probiotic cultures. [Projekat Ministarstva nauke Republike Srbije, br. 046010

INHIBITION OF STAPHYLOCOCCUS AUREUS BY LACTIC ACID BACTERIA AND / OR BIFIDOBACTERIUM LACTIS DURING MILK FERMENTATION AND STORAGE

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Khalaf S. Al-Delaimy

2013-02-01

Full Text Available Survival and inhibition of Staphylococcus aureus by the lactic acid bacteria (LAB starter culture (Sterptococcus thermophillus and Lactobacillus delbrukii subsp. bulgaricus and/ or probiotic bacteria Bifidobacterium lactis during milk fermentation to yoghurt and storage up to 12 days was studied. Adding S. aureus (initial count log 6.64/ ml with LAB (initial count log 6.8/ ml in milk during yoghurt processing and storage resulted in no significant change in the counts of both S. aureus and LAB during fermentation period of 4 hrs at 45° C. A steady decrease in S. aureus count during storage at 25° C and 4° C was observed reaching a complete (100 % inhibition after 9 and 12 days, respectively, with no significant increase in LAB count. Adding S. aureus (initial count log 6.62/ ml with B. lactis (initial count log 6.83/ ml in milk for 4 hr at45° C, no significant changes in the counts of both bacteria were found. After storage at 25° C and at 4° C a sharp decline in the S. aureus count with a 100 % inhibition after 6 and 9 days with approximately two log and one log increase in B. lactis counts consecutively. In general similar result was observed when adding S. aureus together with LAB and B. lactis in milk during fermentation and storage. pH values decreased during milk fermentation and storage from initially 6.55-6.64 to around 4 in most milk samples. The results of this study show that S. aureus was completely inhibited by LAB and/or B. lactis after milk fermentation to yoghurt and storage at room temperature and refrigeration for 6-9 days. It is therefore recommended to add the probiotic B. lactis with LAB to milk for yoghurt processing.

Screening for crude oil degrading bacteria in liquid organic waste (effluent samples)

International Nuclear Information System (INIS)

Akpe, A.R.

2014-01-01

The screening for crude oil degrading bacteria in some liquid organic wastes (cassava mill effluents, rubber effluents and oil palm mill effluents) was carried out. Hydrocarbon utilising bacteria were isolated on mineral salt agar using vapour phase technique. The samples yielded 20 bacterial isolates from 13 different genera. Cassava mill effluent and rubber effluent had the highest number (7), while oil palm effluent had the least number (6) of bacterial isolates. The isolates that had the highest occurrence (occurring in all samples) were Pseudomonas aeruginosa and Escherichia coli. Of these 13 genera 9 were gram negative, while only 4 were gram positive. The total heterotrophic bacterial (THB) count and total hydrocarbon utilisers (THU) from all the effluent samples ranged from 3.0 * 10/sup 4/ to 6.0* 10/sup 7/ cfu/mL and 2.3 *10/sup 2/ to 4.2*10/sup 3/ cfu/mL, respectively. The counts of hydrocarbon utilisers were obviously lower than the heterotrophic counts, although the differences in counts were found to be statistically non-significant (P > 0.05). Rubber effluents and oil palm mill effluents had the highest number of hydrocarbon utilisers with three isolates each. The active hydrocarbon utilisers encountered in this study included Serratia marscescens, Bacillus cereus, P. aeruginosa, Enterobacter aerogenes and Bacillus subtilis. Presence of nutrients and crude oil degrading bacteria in these effluents suggests that these effluents can be used to enhance bioremediation through their use as biostimulation and bioaugmentation agents. (author)

A Sortase A-Immobilized Mesoporous Hollow Carbon Sphere-Based Biosensor for Detection of Gram-Positive Bacteria

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Wang, Hongsu; Luo, Ruiping; Chen, Yang; Si, Qi; Niu, Xiaodi

2018-05-01

A sensor based on mesoporous carbon materials immobilized with sortase A (SrtA) for determination of Staphylococcus aureus (S. aureus) is reported. To prepare the biosensor, we first synthesized carboxyl-functionalized mesoporous hollow carbon spheres, then applied them as carriers for immobilization of SrtA. Based on the catalytic mechanism of SrtA, a highly sensitive, inexpensive, and rapid method was developed for S. aureus detection. The sensor showed a linear response in the bacterial concentration range of 0.125 × 102 colony-forming units (CFU) mL-1 to 2.5 × 102 CFU mL-1, with detection limit as low as 9.0 CFU mL-1. The method was successfully used for quantitative detection of S. aureus in whole milk samples, giving results similar to experimental results obtained from the plate counting method. This biosensor could also be used to detect other Gram-positive bacteria that secrete SrtA.

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

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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.

Developments in microchannel plate detectors for imaging x-ray astronomy

International Nuclear Information System (INIS)

Fraser, G.W.; Whiteley, M.J.; Pearson, J.F.

1985-01-01

The authors present new results in four areas of microchannel plate (MCP) X-ray detector operation. The performance in pulse counting mode of MCPs with 8 micron channel diameters is reported. The effects on MCP quantum detection efficiency and energy discrimination of multiple CsI coatings are described. A new mode of operation of two-stage multipliers is evaluated. Replacing the conventional electron-accelerating inter-plate potential difference by a retarding field is shown to result in definite advantages with regard to X-ray energy discrimination and detector lifetime. The source of the MCP internal background is discussed

LEVELS AND TYPES OF AEROBIC SPORE FORMING BACTERIA ...

African Journals Online (AJOL)

The four companies whose packaged product were studied had an average plate total spore counts as follows: Company A=6.2x 103; Company B= 3.1x 104; Company C= 6.0x 104 and Company D= 3.1x102 colony forming units per gram, respectively. Identification tests showed that among the aerobic spore formers were ...

Impact of manure fertilization on the abundance of antibiotic-resistant bacteria and frequency of detection of antibiotic resistance genes in soil and on vegetables at harvest.

Science.gov (United States)

Marti, Romain; Scott, Andrew; Tien, Yuan-Ching; Murray, Roger; Sabourin, Lyne; Zhang, Yun; Topp, Edward

2013-09-01

Consumption of vegetables represents a route of direct human exposure to bacteria found in soil. The present study evaluated the complement of bacteria resistant to various antibiotics on vegetables often eaten raw (tomato, cucumber, pepper, carrot, radish, lettuce) and how this might vary with growth in soil fertilized inorganically or with dairy or swine manure. Vegetables were sown into field plots immediately following fertilization and harvested when of marketable quality. Vegetable and soil samples were evaluated for viable antibiotic-resistant bacteria by plate count on Chromocult medium supplemented with antibiotics at clinical breakpoint concentrations. DNA was extracted from soil and vegetables and evaluated by PCR for the presence of 46 gene targets associated with plasmid incompatibility groups, integrons, or antibiotic resistance genes. Soil receiving manure was enriched in antibiotic-resistant bacteria and various antibiotic resistance determinants. There was no coherent corresponding increase in the abundance of antibiotic-resistant bacteria enumerated from any vegetable grown in manure-fertilized soil. Numerous antibiotic resistance determinants were detected in DNA extracted from vegetables grown in unmanured soil. A smaller number of determinants were additionally detected on vegetables grown only in manured and not in unmanured soil. Overall, consumption of raw vegetables represents a route of human exposure to antibiotic-resistant bacteria and resistance determinants naturally present in soil. However, the detection of some determinants on vegetables grown only in freshly manured soil reinforces the advisability of pretreating manure through composting or other stabilization processes or mandating offset times between manuring and harvesting vegetables for human consumption.

Screening and biological characteristics of fufenozide degrading bacteria

Science.gov (United States)

Xu, Chenhao; Gong, Mingfu; Guan, Qinlan; Deng, Xia; Deng, Hongyan; Huang, Jiao

2018-04-01

Fufenozide was a novel pesticide for the control of Lepidoptera pests, which was highly toxic to silkworm. Fufenozide-contaminated soil samples were collected and the bacteria that degrade fufenozide were isolated and screened by selective medium. The colony characteristics, cell characteristics and degradation characteristics in different concentrations fufenozide of the fufenozide degrading bacteria were studied. The results indicated that seven strains of fufenozide degradeing bacteria, named as DDH01, DDH03, DDH04, DDH04, DDH05, DDH07 and DDH07 respectively, were isolated from soil contaminated with fufenozide. DDH01, DDH02, DDH04 and DDH05 of seven fufenozide degrading bacteria, was gram-positive bacteria, and DDH03, DDH06 and DDH07 was gram-negative bacteria. All of seven strains of fufenozide degrading bacteria were not spores, weeks flagella, rod-shaped bacteria. DDH06 and DDH07 had capsules, and the remaining five strains had not capsule. The colonies formed by seven strains of fufenozide degradation bacteria on beef extract peptone medium plate were milky white colonies with irregular edges, thinner lawn, smaller colony with smooth surface. The growth of 7 strains of fufenozide degradation bacteria was significantly affected by the concentration of fufenozide, All of 7 strains grown in the range from 0.00025 g/mL to 1 g/mL of 10% fufenozide suspension. DDH2 was the best among the 7 strains of fufenozide degrading bacteria grown in 10% fufenozide suspension medium.

Extracellular deoxyribonuclease production by periodontal bacteria.

Science.gov (United States)

Palmer, L J; Chapple, I L C; Wright, H J; Roberts, A; Cooper, P R

2012-08-01

Whilst certain bacteria have long been known to secrete extracellular deoxyribonuclease (DNase), the purpose in microbial physiology was unclear. Recently, however, this enzyme has been demonstrated to confer enhanced virulence, enabling bacteria to evade the host's immune defence of extruded DNA/chromatin filaments, termed neutrophil extracellular traps (NETs). As NETs have recently been identified in infected periodontal tissue, the aim of this study was to screen periodontal bacteria for extracellular DNase activity. To determine whether DNase activity was membrane bound or secreted, 34 periodontal bacteria were cultured in broth and on agar plates. Pelleted bacteria and supernatants from broth cultures were analysed for their ability to degrade DNA, with relative activity levels determined using an agarose gel electrophoresis assay. Following culture on DNA-supplemented agar, expression was determined by the presence of a zone of hydrolysis and DNase activity related to colony size. Twenty-seven bacteria, including red and orange complex members Porphyromonas gingivalis, Tannerella forsythia, Fusobacterium nucleatum, Parvimonas micra, Prevotella intermedia, Streptococcus constellatus, Campylobacter rectus and Prevotella nigrescens, were observed to express extracellular DNase activity. Differences in DNase activity were noted, however, when bacteria were assayed in different culture states. Analysis of the activity of secreted DNase from bacterial broth cultures confirmed their ability to degrade NETs. The present study demonstrates, for the first time, that DNase activity is a relatively common property of bacteria associated with advanced periodontal disease. Further work is required to determine the importance of this bacterial DNase activity in the pathogenesis of periodontitis. © 2011 John Wiley & Sons A/S.

Optimasi Konsentrasi Fruktooligosakarida untuk Meningkatkan Pertumbuhan Bakteri Asam Laktat Starter Yoghurt (CONCENTRATION OPTIMIZATION OF FRUCTOOLIGOSACCHARIDES TO INCREASE GROWTH OF LACTIC ACID BACTERIA YOGHURT STARTER

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Raden Haryo Bimo Setiarto

2017-09-01

Full Text Available Fructooligosaccharides are prebiotic source that widely used in food products, such as: fermented milk and infant formula. Prebiotics are food components that cannot be digested in the digestive tract enzymatically. However, they can be fermented by probiotic bacteria in the colon. This study aimed to determine the optimum concentrations of fructooligosaccharides in order to increase the growth of lactic acid bacteria yogurt starter (Lactobacillus acidophillus, Lactobacillus bulgaricus, Streptococcus thermophillus. Optimation concentration of fructooligosaccharides on the growth of Lactobacillus acidophilus, Lactobacillus bulgaricus, Streptococcus thermophillus can be determined based on OD (optical density, TPC (Total Plate Count, total lactic acid content and pH value. Suplementation of fructooligosaccharides 1 % (w/v on the media MRSB increased significantly the growth of L. acidophilus, L.bulgaricus, S. thermophilus. Furthermore, L. acidophilus, L. bulgaricus and S. thermophilus experienced exponential growth phase during incubation period from 6 to 18 hours. Fermentation of L. acidophilus, L. bulgaricus, S. thermophilus in MRSB medium supplemented by fructooligosaccharides decreased the pH value of the formation of organic acids from 6.00 to 4.00. ABSTRAK Fruktooligosakarida adalah sumber prebiotik yang banyak digunakan dalam produk pangan olahan seperti susu fermentasi dan susu formula. Prebiotik adalah komponen bahan pangan fungsional yang tidak dapat dicerna di dalam saluran pencernaan secara enzimatik sehingga akan difermentasi oleh bakteri probiotik dalam usus besar. Penelitian ini bertujuan menentukan konsentrasi optimum fruktooligosakarida untuk meningkatkan pertumbuhan bakteri asam laktat starter yoghurt (Lactobacillus acidophillus, Lactobacillus bulgaricus, Streptococcus thermophillus. Konsentrasi optimum fruktooligosakarida pada pertumbuhan Lactobacillus acidophilus, Lactobacillus bulgaricus, Streptococcus thermophillus dapat

How bacterial cell division might cheat turgor pressure - a unified mechanism of septal division in Gram-positive and Gram-negative bacteria.

Science.gov (United States)

Erickson, Harold P

2017-08-01

An important question for bacterial cell division is how the invaginating septum can overcome the turgor force generated by the high osmolarity of the cytoplasm. I suggest that it may not need to. Several studies in Gram-negative bacteria have shown that the periplasm is isoosmolar with the cytoplasm. Indirect evidence suggests that this is also true for Gram-positive bacteria. In this case the invagination of the septum takes place within the uniformly high osmotic pressure environment, and does not have to fight turgor pressure. A related question is how the V-shaped constriction of Gram-negative bacteria relates to the plate-like septum of Gram-positive bacteria. I collected evidence that Gram-negative bacteria have a latent capability of forming plate-like septa, and present a model in which septal division is the basic mechanism in both Gram-positive and Gram-negative bacteria. © 2017 WILEY Periodicals, Inc.

Total resistance of native bacteria as an indicator of changes in the water environment

Energy Technology Data Exchange (ETDEWEB)

Harnisz, Monika [Department of Environmental Microbiology, University of Warmia and Mazury in Olsztyn, Prawocheńskiego 1, 10-957 Olsztyn (Poland)

2013-03-15

This study analyzes changes in the total (intrinsic and acquired) resistance of autochthonous bacteria in a river which is a receiver of treated wastewater. In the analyzed samples, tetracycline contamination levels were low and characteristic of surface water bodies. An increase in the populations of tetracycline-resistant and fluoroquinolone-resistant microorganisms was noted in downstream river water samples in comparison with upstream river water samples, but the above trend was not observed in bacteria resistant to macrolides and β-lactams. The counts of doxycycline-resistant bacteria (DOX{sup R}) were significantly correlated with doxycycline levels. The minimum inhibitory concentrations (MICs) for doxycycline in DOX{sup R} isolates were higher in downstream river water than in upstream river water samples. The discharge of treated wastewater had no effect on the multi-drug resistance of oxytetracycline-resistant and doxycycline-resistant isolates. The results of the experiment indicate that the presence of doxycycline-resistant bacteria is a robust indicator of anthropogenic stress in river water. -- Highlights: ► The total resistance of native bacteria in river which is a receiver of treated wastewater was analyzed. ► Tetracyclines contamination levels were low. ► The counts of doxycycline-resistant bacteria were correlated with doxycycline levels. -- The presence of doxycycline-resistant bacteria in rivers can be a robust indicator of anthropogenic stress.

Total resistance of native bacteria as an indicator of changes in the water environment

International Nuclear Information System (INIS)

Harnisz, Monika

2013-01-01

This study analyzes changes in the total (intrinsic and acquired) resistance of autochthonous bacteria in a river which is a receiver of treated wastewater. In the analyzed samples, tetracycline contamination levels were low and characteristic of surface water bodies. An increase in the populations of tetracycline-resistant and fluoroquinolone-resistant microorganisms was noted in downstream river water samples in comparison with upstream river water samples, but the above trend was not observed in bacteria resistant to macrolides and β-lactams. The counts of doxycycline-resistant bacteria (DOX R ) were significantly correlated with doxycycline levels. The minimum inhibitory concentrations (MICs) for doxycycline in DOX R isolates were higher in downstream river water than in upstream river water samples. The discharge of treated wastewater had no effect on the multi-drug resistance of oxytetracycline-resistant and doxycycline-resistant isolates. The results of the experiment indicate that the presence of doxycycline-resistant bacteria is a robust indicator of anthropogenic stress in river water. -- Highlights: ► The total resistance of native bacteria in river which is a receiver of treated wastewater was analyzed. ► Tetracyclines contamination levels were low. ► The counts of doxycycline-resistant bacteria were correlated with doxycycline levels. -- The presence of doxycycline-resistant bacteria in rivers can be a robust indicator of anthropogenic stress

A study of bacteria, fungi and biomass in particulate matter in ambient air of Khorramabad during summer and autumn 2012

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hatam Godini

2015-05-01

Full Text Available Introduction: Particulate matter refers to the combination of atmospheric pollutants that a portion of this particulate is bioaerosol. The aim of this study was the evaluation of bacteria, fungi and biomass in particulate matter in ambient air of Khorramabad during summer and autumn 2012. Materials and Methods: This study was a cross sectional study that conducted in Khorramabad city during summer and fall 2012. Sampling has been done via high-volume sampler. The special cultures were used for cultivation and determination of fungal and Heterotrophic Plate Count (HPC (and Bradford method were used to determine bacteria and protein as biomass indicator, respectively. Relationship between these variables with metrological parameters was evaluated too. Results: The highest PM10 in July (257.18 µg/m3 and lowest in September (92.45 µg/m3 had been recorded. The highest amount of bacteria and fungi were measured as monthly in November (605 No/m3 and December (120 No/m3, respectively. The highest of protein concentration was measured in August, September and December (27-30 µg/m3. With the increase in PM10, biomass concentration in the air showed a meaningful increase. Conclusion: Biomass concentration in the air increased with increasing PM10 but it had no significant effect on the concentration of bacteria and fungi in the air. Meteorological factors such as temperature, humidity, wind speed, solar radiation and the amount of exposure time had a significant impact on bioaerosol concentrations in the air.

Visualizing aquatic bacteria by light and transmission electron microscopy.

Science.gov (United States)

Silva, Thiago P; Noyma, Natália P; Duque, Thabata L A; Gamalier, Juliana P; Vidal, Luciana O; Lobão, Lúcia M; Chiarini-Garcia, Hélio; Roland, Fábio; Melo, Rossana C N

2014-01-01

The understanding of the functional role of aquatic bacteria in microbial food webs is largely dependent on methods applied to the direct visualization and enumeration of these organisms. While the ultrastructure of aquatic bacteria is still poorly known, routine observation of aquatic bacteria by light microscopy requires staining with fluorochromes, followed by filtration and direct counting on filter surfaces. Here, we used a new strategy to visualize and enumerate aquatic bacteria by light microscopy. By spinning water samples from varied tropical ecosystems in a cytocentrifuge, we found that bacteria firmly adhere to regular slides, can be stained by fluorochoromes with no background formation and fast enumerated. Significant correlations were found between the cytocentrifugation and filter-based methods. Moreover, preparations through cytocentrifugation were more adequate for bacterial viability evaluation than filter-based preparations. Transmission electron microscopic analyses revealed a morphological diversity of bacteria with different internal and external structures, such as large variation in the cell envelope and capsule thickness, and presence or not of thylakoid membranes. Our results demonstrate that aquatic bacteria represent an ultrastructurally diverse population and open avenues for easy handling/quantification and better visualization of bacteria by light microscopy without the need of filter membranes.

The use of flow cytometry to accurately ascertain total and viable counts of Lactobacillus rhamnosus in chocolate.

Science.gov (United States)

Raymond, Yves; Champagne, Claude P

2015-04-01

The goals of this study were to evaluate the precision and accuracy of flow cytometry (FC) methodologies in the evaluation of populations of probiotic bacteria (Lactobacillus rhamnosus R0011) in two commercial dried forms, and ascertain the challenges in enumerating them in a chocolate matrix. FC analyses of total (FC(T)) and viable (FC(V)) counts in liquid or dried cultures were almost two times more precise (reproducible) than traditional direct microscopic counts (DCM) or colony forming units (CFU). With FC, it was possible to ascertain low levels of dead cells (FC(D)) in fresh cultures, which is not possible with traditional CFU and DMC methodologies. There was no interference of chocolate solids on FC counts of probiotics when inoculation was above 10(7) bacteria per g. Addition of probiotics in chocolate at 40 °C resulted in a 37% loss in viable cells. Blending of the probiotic powder into chocolate was not uniform which raised a concern that the precision of viable counts could suffer. FCT data can serve to identify the correct inoculation level of a sample, and viable counts (FCV or CFU) can subsequently be better interpreted. Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.

Antimicrobial-resistant bacteria in wild game in Slovenia

Science.gov (United States)

Križman, M.; Kirbiš, A.; Jamnikar-Ciglenečki, U.

2017-09-01

Wildlife is usually not exposed to clinically-used antimicrobial agents but can acquire antimicrobial resistance throughout contact with humans, domesticated animals and environments. Samples of faeces from intestines (80 in total) were collected from roe deer (52), wild boars (11), chamois (10) red deer (6) and moufflon (1). After culture on ChromID extended spectrum β-lactamase (ESBL) plates to select for growth of ESBL-producing bacteria, 25 samples produced bacterial colonies for further study. Six species of bacteria were identified from the 25 samples: Stenotrophomonas maltophilia, Serratia fonticola, Stenotrophomonas nitritireducens, Enterococcus faecium, Enterococcus faecalis and Escherichia coli. Two ESBL enzymes were amplified from group TEM and three from group CTX-M-1. Undercooked game meat and salami can be a source of resistant bacteria when animals are not eviscerated properly.

Broth and agar hop-gradient plates used to evaluate the beer-spoilage potential of Lactobacillus and Pediococcus isolates.

Science.gov (United States)

Haakensen, M; Schubert, A; Ziola, B

2009-03-15

Identification of the beer-spoilage Lactobacillus and Pediococcus bacteria has largely taken two approaches; identification of spoilage-associated genes or identification of specific species of bacteria regardless of ability to grow in beer. The problem with these two approaches is that they are either overly inclusive (i.e., detect all bacteria of a given species regardless of spoilage potential) or overly selective (i.e., rely upon individual, putative spoilage-associated genes). Our goal was to design a method to assess the ability of Lactobacillus and Pediococcus to spoil beer that is independent of speciation or genetic background. In searching for a method by which to differentiate between beer-spoilage bacteria and bacteria that cannot grow in beer, we explored the ability of lactobacilli and pediococci isolates to grow in the presence of varying concentrations of hop-compounds and ethanol in broth medium versus on agar medium. The best method for differentiating between bacteria that can grow in beer and bacteria that do not pose a threat as beer-spoilage organisms was found to be a hop-gradient agar plate containing ethanol. This hop-gradient agar plate technique provides a rapid and simple solution to the dilemma of assessing the ability of Lactobacillus and Pediococcus isolates to grow in beer, and provides new insights into the different strategies used by these bacteria to survive under the stringent conditions of beer.

Cytotoxicity But No Mutagenicity In Bacteria With Externally Generated Singlet Oxygen

Science.gov (United States)

Midden, W. Robert; Dahl, Thomas A.; Hartman, Philip E.

1988-02-01

Singlet oxygen is believed to be an important intermediate responsible for the cytotoxicity of HpD phototherapy. It has been recognized as a possible intermediate in photosensitization for more than 20 years. However, it has been difficult to obtain conclusive evidence of its biological characteristics in the past because most of the methods available for its generation that are compatible with biological systems also generate other reactive intermediates whose effects are difficult to distinguish from singlet oxygen. We have used a recently devised separated-surface-sensi-tizer (S-S-S) system for singlet oxygen generation' to measure the cytotoxicity and mutagenicity of singlet oxygen in bacteria. The S-S-S system employs rose bengal as a sensitizer immobilized on one surface of a glass plate. The glass plate is placed sensitizer-side down a small distance (plate is illuminated from above to generate singlet oxygen at the surface of the sensitizer. The singlet oxygen thus generated can diffuse the short dis-tance to the surface of the membrane to react with the bacteria. Because of the short lifetime of singlet oxygen in air, increasing the distance between the sensitizer and the membrane causes a decline in the amount of singlet oxygen reaching the membrane according to a function derived from the Einstein-Smoluchowski equation for net displacement by diffusion. Plotting the log of the effect measured (e.g., cytotoxicity) vs. the square of the distance gives a straight line. The slope of this line can be used to calculate the gas phase half life of the intermediate responsible for the observed effects. We have found that bacteria are rapidly killed in the illuminated S-S-S system and that the gas phase half life of the agent responsible for cell killing is the same as that of singlet oxygen. This observation and other simple chemical tests have conclusively estab-lished that singlet oxygen is responsible for the cytotoxicity observed with bacteria. Dosimetry

Mycobacterium tuberculosis from chronic murine infections that grows in liquid but not on solid medium

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Mitchison Denis A

2004-11-01

Full Text Available Abstract Background Old, stationary cultures of Mycobacterium tuberculosis contain a majority of bacteria that can grow in broth cultures but cannot grow on solid medium plates. These may be in a non-replicating, dormant growth phase. We hypothesised that a similar population might be present in chronic, murine tuberculosis. Methods Estimates of the numbers of viable M. tuberculosis, strain H37Rv, in the spleens and lungs of mice in a 7-day acute infection and in a 10-month chronic infection were made by conventional plate counts and, as broth counts, by noting presence or absence of growth in serial replicate dilutions in liquid medium. Results Plate and broth counts in 6 mice gave similar mean values in the acute infection, 7 days after infection. However, the broth counts were much higher in 36 mice with a chronic infection at 10 months. Broth counts averaged 5.290 log10 cfu /organ from spleens and 5.523 log10 cfu/organ from lungs, while plate counts were 3.858 log10 cfu/organ from spleens and 3.662 log10 cfu/organ from lungs, indicating that the total bacterial population contained only 3.7% bacilli in spleens and 1.4% bacilli in lungs, capable of growth on plates. Conclusion The proportion growing on plates might be a measure of the "dormancy" of the bacilli equally applicable to cultural and animal models.

[Study on modeling method of total viable count of fresh pork meat based on hyperspectral imaging system].

Science.gov (United States)

Wang, Wei; Peng, Yan-Kun; Zhang, Xiao-Li

2010-02-01

Once the total viable count (TVC) of bacteria in fresh pork meat exceeds a certain number, it will become pathogenic bacteria. The present paper is to explore the feasibility of hyperspectral imaging technology combined with relevant modeling method for the prediction of TVC in fresh pork meat. For the certain kind of problem that has remarkable nonlinear characteristic and contains few samples, as well as the problem that has large amount of data used to express the information of spectrum and space dimension, it is crucial to choose a logical modeling method in order to achieve good prediction result. Based on the comparative result of partial least-squares regression (PLSR), artificial neural networks (ANNs) and least square support vector machines (LS-SVM), the authors found that the PLSR method was helpless for nonlinear regression problem, and the ANNs method couldn't get approving prediction result for few samples problem, however the prediction models based on LS-SVM can give attention to the little training error and the favorable generalization ability as soon as possible, and can make them well synchronously. Therefore LS-SVM was adopted as the modeling method to predict the TVC of pork meat. Then the TVC prediction model was constructed using all the 512 wavelength data acquired by the hyperspectral imaging system. The determination coefficient between the TVC obtained with the standard plate count for bacterial colonies method and the LS-SVM prediction result was 0.987 2 and 0.942 6 for the samples of calibration set and prediction set respectively, also the root mean square error of calibration (RMSEC) and the root mean square error of prediction (RMSEP) was 0.207 1 and 0.217 6 individually, and the result was considerably better than that of MLR, PLSR and ANNs method. This research demonstrates that using the hyperspectral imaging system coupled with the LS-SVM modeling method is a valid means for quick and nondestructive determination of TVC of pork

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

Science.gov (United States)

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.

MALDI-TOF mass spectrometry for early identification of bacteria grown in blood culture bottles.

Science.gov (United States)

Zabbe, Jean-Benoît; Zanardo, Laura; Mégraud, Francis; Bessède, Emilie

2015-08-01

This note reports an interesting way to rapidly identify bacteria grown from blood culture bottles. Chocolate agar plates were inoculated with 1 drop of the positive blood bottle medium. After a 3-hour incubation, the growth veil was submitted to MALDI-TOF mass spectrometry: 77% of the bacteria present have been correctly identified. Copyright © 2015 Elsevier B.V. All rights reserved.

Estimates of microbial quality and concentration of copper in distributed drinking water are highly dependent on sampling strategy.

Science.gov (United States)

Lehtola, Markku J; Miettinen, Ilkka T; Hirvonen, Arja; Vartiainen, Terttu; Martikainen, Pertti J

2007-12-01

The numbers of bacteria generally increase in distributed water. Often household pipelines or water fittings (e.g., taps) represent the most critical location for microbial growth in water distribution systems. According to the European Union drinking water directive, there should not be abnormal changes in the colony counts in water. We used a pilot distribution system to study the effects of water stagnation on drinking water microbial quality, concentration of copper and formation of biofilms with two commonly used pipeline materials in households; copper and plastic (polyethylene). Water stagnation for more than 4h significantly increased both the copper concentration and the number of bacteria in water. Heterotrophic plate counts were six times higher in PE pipes and ten times higher in copper pipes after 16 h of stagnation than after only 40 min stagnation. The increase in the heterotrophic plate counts was linear with time in both copper and plastic pipelines. In the distribution system, bacteria originated mainly from biofilms, because in laboratory tests with water, there was only minor growth of bacteria after 16 h stagnation. Our study indicates that water stagnation in the distribution system clearly affects microbial numbers and the concentration of copper in water, and should be considered when planning the sampling strategy for drinking water quality control in distribution systems.

Results from raw milk microbiological tests do not predict the shelf-life performance of commercially pasteurized fluid milk.

Science.gov (United States)

Martin, N H; Ranieri, M L; Murphy, S C; Ralyea, R D; Wiedmann, M; Boor, K J

2011-03-01

Analytical tools that accurately predict the performance of raw milk following its manufacture into commercial food products are of economic interest to the dairy industry. To evaluate the ability of currently applied raw milk microbiological tests to predict the quality of commercially pasteurized fluid milk products, samples of raw milk and 2% fat pasteurized milk were obtained from 4 New York State fluid milk processors for a 1-yr period. Raw milk samples were examined using a variety of tests commonly applied to raw milk, including somatic cell count, standard plate count, psychrotrophic bacteria count, ropy milk test, coliform count, preliminary incubation count, laboratory pasteurization count, and spore pasteurization count. Differential and selective media were used to identify groups of bacteria present in raw milk. Pasteurized milk samples were held at 6°C for 21 d and evaluated for standard plate count, coliform count, and sensory quality throughout shelf-life. Bacterial isolates from select raw and pasteurized milk tests were identified using 16S ribosomal DNA sequencing. Linear regression analysis of raw milk test results versus results reflecting pasteurized milk quality consistently showed low R(2) values (tests and results from tests used to evaluate pasteurized milk quality. Our findings suggest the need for new raw milk tests that measure the specific biological barriers that limit shelf-life and quality of fluid milk products. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

UV inactivation of pathogenic and indicator microorganisms

International Nuclear Information System (INIS)

Chang, J.C.; Ossoff, S.F.; Lobe, D.C.; Dorfman, M.H.; Dumais, C.M.; Qualls, R.G.; Johnson, J.D.

1985-01-01

Survival was measured as a function of the dose of germicidal UV light for the bacteria Escherichia coli, Salmonella typhi, Shigella sonnei, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis spores, the enteric viruses poliovirus type 1 and simian rotavirus SA11, the cysts of the protozoan Acanthamoeba castellanii, as well as for total coliforms and standard plate count microorganisms from secondary effluent. The doses of UV light necessary for a 99.9% inactivation of the cultured vegetative bacteria, total coliforms, and standard plate count microorganisms were comparable. However, the viruses, the bacterial spores, and the amoebic cysts required about 3 to 4 times, 9 times, and 15 times, respectively, the dose required for E. coli. These ratios covered a narrower relative dose range than that previously reported for chlorine disinfection of E. coli, viruses, spores, and cysts

UV inactivation of pathogenic and indicator microorganisms

Energy Technology Data Exchange (ETDEWEB)

Chang, J.C.; Ossoff, S.F.; Lobe, D.C.; Dorfman, M.H.; Dumais, C.M.; Qualls, R.G.; Johnson, J.D.

1985-06-01

Survival was measured as a function of the dose of germicidal UV light for the bacteria Escherichia coli, Salmonella typhi, Shigella sonnei, Streptococcus faecalis, Staphylococcus aureus, and Bacillus subtilis spores, the enteric viruses poliovirus type 1 and simian rotavirus SA11, the cysts of the protozoan Acanthamoeba castellanii, as well as for total coliforms and standard plate count microorganisms from secondary effluent. The doses of UV light necessary for a 99.9% inactivation of the cultured vegetative bacteria, total coliforms, and standard plate count microorganisms were comparable. However, the viruses, the bacterial spores, and the amoebic cysts required about 3 to 4 times, 9 times, and 15 times, respectively, the dose required for E. coli. These ratios covered a narrower relative dose range than that previously reported for chlorine disinfection of E. coli, viruses, spores, and cysts.

Azoreductase activity of anaerobic bacteria isolated from human intestinal microflora.

Science.gov (United States)

Rafii, F; Franklin, W; Cerniglia, C E

1990-07-01

A plate assay was developed for the detection of anaerobic bacteria that produce azoreductases. With this plate assay, 10 strains of anaerobic bacteria capable of reducing azo dyes were isolated from human feces and identified as Eubacterium hadrum (2 strains), Eubacterium spp. (2 species), Clostridium clostridiiforme, a Butyrivibrio sp., a Bacteroides sp., Clostridium paraputrificum, Clostridium nexile, and a Clostridium sp. The average rate of reduction of Direct Blue 15 dye (a dimethoxybenzidine-based dye) in these strains ranged from 16 to 135 nmol of dye per min per mg of protein. The enzymes were inactivated by oxygen. In seven isolates, a flavin compound (riboflavin, flavin adenine dinucleotide, or flavin mononucleotide) was required for azoreductase activity. In the other three isolates and in Clostridium perfringens, no added flavin was required for activity. Nondenaturing polyacrylamide gel electrophoresis showed that each bacterium expressed only one azoreductase isozyme. At least three types of azoreductase enzyme were produced by the different isolates. All of the azoreductases were produced constitutively and released extracellularly.

A six-well plate method: less laborious and effective method for cultivation of obligate anaerobic microorganisms.

Science.gov (United States)

Nakamura, Kohei; Tamaki, Hideyuki; Kang, Myung Suk; Mochimaru, Hanako; Lee, Sung-Taik; Nakamura, Kazunori; Kamagata, Yoichi

2011-01-01

We developed a simple, less laborious method to cultivate and isolate obligate anaerobic microorganisms using a six-well plate together with the AnaeroPack System, designated as the six-well plate method. The cultivation efficiency of this method, based on colony-forming units, colony formation time, and colony size, was evaluated with four authentic obligate anaerobes (two methanogenic archaea and two sulfate-reducing bacteria). The method was found to be comparable to or even better than the roll tube method, a technique that is commonly used at present for the cultivation of obligate anaerobes. Further experiments using 21 representative obligate anaerobes demonstrated that all examined anaerobes (11 methanogens, 5 sulfate- or thiosulfate-reducing bacteria, and 5 syntrophs) could form visible colonies on the six-well plate and that these colonies could be successfully subcultured in fresh liquid media. Using this method, an unidentified sulfate-reducing bacterium was successfully isolated from an environmental sample.

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

Science.gov (United States)

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 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.

Influence of disturbances on bacteria level in an operating room

DEFF Research Database (Denmark)

Brohus, Henrik; Hyldig, Mikkel; Kamper, Simon

2008-01-01

In operating rooms great effort is manifested to reduce the bacteria level in order to decrease the risk of infections. The main source of bacteria is the staff and the patient, thus, the resulting bacteria concentration is roughly speaking a combination of the ventilation system and the emission...... from the occupants. This study investigates the influence of two main disturbances in an operating room namely the door opening during the operation and the activity level of the staff. It is found that the frequent door opening in this case does not cause significant transport of air from outside...... the operating room to the wound area of the patient. However, a significant influence of the activity level on the bacteria emission and concentration is found. Counting the number of persons in an operating room to estimate the bacteria source strength is not sufficient, the corresponding activity level must...

The application of flow cytometry and fluorescent probe technology for detection and assessment of viability of plant pathogenic bacteria

NARCIS (Netherlands)

Chitarra, L.G.; Bulk, van den R.W.

2003-01-01

Conventional methods to detect and assess the viability of plant pathogenic bacteria are usually based on plating assays or serological techniques. Plating assays provide information about the number of viable cells, expressed as colony-forming units, but are time-consuming and laborious.

Effectiveness of a pre-procedural mouthwash in reducing bacteria in dental aerosols: randomized clinical trial

Directory of Open Access Journals (Sweden)

Belén RETAMAL-VALDES

2017-03-01

Full Text Available Abstract The aim of this randomized, single blinded clinical trial was to evaluate the effect of a pre-procedural mouthwash containing cetylpyridinium chloride (CPC, zinc lactate (Zn and sodium fluoride (F in the reduction of viable bacteria in oral aerosol after a dental prophylaxis with ultrasonic scaler. Sixty systemically healthy volunteers receiving dental prophylaxis were randomly assigned to one of the following experimental groups (15 per group: (i rinsing with 0.075% CPC, 0.28% Zn and 0.05% F (CPC+Zn+F, (ii water or (iii 0.12% chlorhexidine digluconate (CHX, and (iv no rinsing. Viable bacteria were collected from different locations in the dental office on enriched TSA plates and anaerobically incubated for 72 hours. The colonies were counted and species were then identified by Checkerboard DNA–DNA Hybridization. The total number of colony-forming units (CFUs detected in the aerosols from volunteers who rinsed with CPC+Zn+F or CHX was statistically significantly (p<0.05 lower than of those subjects who did not rinse or who rinsed with water. When all locations were considered together, the aerosols from the CPC+Zn+F and CHX groups showed, respectively, 70% and 77% fewer CFUs than those from the No Rinsing group and 61% and 70% than those from the Water group. The mean proportions of bacterial species from the orange complex were statistically significantly (p<0.05 lower in aerosols from the CPC+Zn+F and CHX groups compared with the others two groups. In conclusion, the mouthwash containing CPC+Zn+F, is effective in reducing viable bacteria in oral aerosol after a dental prophylaxis with ultrasonic scaler.

Abundance and activity of oil-degrading and indigenous bacteria in sediment microcosms

International Nuclear Information System (INIS)

Araujo, R.; Molina, M.; Bachoon, D.

1995-01-01

The responses of bacterial community composition and degradation crude oil to applications of bioremediation products and plant detrital material were investigated in wetlands microcosms. The microcosms were constructed of sieved sediments and operated as tidal marshes. Products included nutrients, organisms, surfactants and combinations thereof; dried ground Spartina was the source of detrital material. Plate count and most probable-number techniques were used to enumerate microbial populations and GC/MS analysis of indicator petroleum hydrocarbons was used to assess oil degradation. Microbial communities were characterized by whole-genome hybridization and specific probes for bacterial groups, including Pseudomonas, Streptomycetes, Vibrio, and sulfate-reducing bacteria. Although the total microbial numbers were similar in all bioremediation treatments, the numbers of oil degraders increased two to three log units in the fertilizer and microbial-degrader-enriched treatments. Oil-degraders comprised the largest fraction of the total population in the treatment amended with microbial degraders, apparently at the expense of indigenous bacteria, as indicated by specific probes. Oil-degraders were also detected in the subsurface in all treatments except the controls. The extent of oil degradation was not consistent with bacterial numbers; only nutrient additions resulted in significantly enhanced degradation of oil. After 1 month of microcosm operation, oil-degraders had increased at least two orders of magnitude in sediment surface layers when oil was added alone or with Spartina detritus, although total bacterial numbers and the number of oil-degraders decreased to near initial levels by 2 months. The peak coincides with bacterial utilization of the alkane fraction of petroleum hydrocarbons

Investigation on adhering properties of dental materials by means of radioactively labelled bacteria

International Nuclear Information System (INIS)

Pfister, W.; Kleinert, P.; Sandig, H.C.; Wutzler, P.; Ruschitschka, A.; Schaefer, U.

1987-01-01

Bacteria of the species Streptococcus mutans were radioactively labelled with 113 In-oxinate. Different dental materials were incubated with the labelled bacteria. Counts per minute of the dental materials could be determined as proportion of the quantity of adhering microorganisms. Silver-palladium-alloy had a lower adherence than silver-tin-alloy. Finest polished alloys had lower adhering properties than unpolished surfaces of materials. (author)

Fly's Eye: a counting camera for thermal neutrons, some applications, problems, and prospects

International Nuclear Information System (INIS)

Davidson, J.B.

1975-01-01

An area detector for thermal neutrons based on image intensification techniques is described and some capabilities and limitations of the detection system are discussed. Among the former are high spatial resolution high instantaneous counting rate, electronic zoom, time-gating, and integration. The detector is limited in that the maximum counting rate for a resolution element is 60 regularly spaced counts per second. Also, the nonuniformity of response over the detector limits the useful size and requires point-by-point calibration. In addition, a higher efficiency for neutron detection would be desirable. Some typical applications of the system are: crystal inspection, neutron magnetic diffraction topography, and searches for temperature induced changes in diffraction patterns. The future application of solid state television sensors and microchannel plate intensifiers to improve the system are briefly mentioned. (U.S.)

Fly's eye: a counting camera for thermal neutrons: some applications, problems, and prospects

International Nuclear Information System (INIS)

Davidson, J.B.

1976-01-01

An area detector for thermal neutrons based on image intensification techniques is described. Some capabilities and limitations of the detection system are discussed. Among the former are high spatial resolution, high instantaneous counting rate, electronic zoom, time-gating, and integration. The detector is limited in that the maximum counting rate for a resolution element is 60 regularly spaced counts per second. Also, the nonuniformity of response over the detector puts a limit on the useful size and necessitates point-by-point calibration. In addition, a higher efficiency for neutron detection would be desirable. Some typical applications of the system are crystal inspection, neutron magnetic diffraction topography, and searches for temperature-induced changes in diffraction patterns. The future application of solid-state television sensors and microchannel-plate intensifiers to improve the system is briefly mentioned

The effect of gamma irradiation on bacteria in stored rice

International Nuclear Information System (INIS)

Kamaruzzaman Sijam.

1987-01-01

The effect of gamma irradiation on bacteria was studied for reducing the total microbial numbers that contaminating raw product under storage. Different storage packages of rice samples were irradiated at various levels of dosage. The results of bacterial isolation, total bacterial count and the isolation of bacterial food pathogenus were discussed. It was observed that the presence of bacteria colonies was suppressed by the presence of yeast and moulds eventhough the number of them decreased as the irradiation dosage levels were increased. (A.J.)

COMPARISON THE NUMBER OF BACTERIA BETWEEN WASHING HANDS USING SOAP AND HAND SANITIZER AS A BACTERIOLOGY LEARNING RESOURCE FOR STUDENTS

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Satya Darmayani

2017-11-01

Full Text Available Hands are the principal carriers of bacterial diseases, therefore very important to know that washing hands with soap or hand sanitizer is highly effective healthy behaviors to reduce bacteria in the palm. This study aimed to determine the total number of bacteria between washing hands with soap and hand sanitizer, also applying the results of these studies as a learning resource in bacteriology. The research design was the true experiment with pretest-posttest control group research design and laboratory examination. Analysis of data using paired t-test and independent sample t-test with α = 0.05. The result using paired t-test obtained t count= 2.48921> t 0.05 (14 = 2.14479 (with liquid soap, obtained t count= 2.32937> t 0.05 (14 = 2.14479 (with hand sanitizer. As for the comparison of the total number of bacteria include washing hands with soap and hand sanitizer using independent samples t-test obtained results there were differences in the total number of bacteria include washing hands with liquid soap and hand sanitizer with t count= 2.23755> t 0.05 ( 13 = 2.16037. That results showed hand sanitizer more effective to reduce the number of bacteria than the liquid soap, that was hand sanitizer 96% and liquid soap by 95%.

Investigations of the effect of electron-beam irradiation on bacteria in sewage sludge

International Nuclear Information System (INIS)

Osterstock, G.

1976-01-01

The effect of electron beams on bacteria was investigated in 2 experimental facilities. A 400 kV electron accelerator was used to irradiate sludge quantities of 10 l and 180 l. The total bacterial count, number of coliform bacteria and, in injected sludge, the relative bacteria density of salmonella were investigated. A dose of 0.5 to 0.75 Mrad was required to reduce coliform bacteria to below the detectable level in 0.1 ml. With a dose of 1.5 Mrad salmonella were reduced by 6 orders of magnitude on the average. In addition, the dependence of the reduction in bacteria on the dose rate as well as on mixing of the irradiation material was investigated. Substantial reproduction of bacteria in digested sludge was found in all cases after the irradiation. (author)

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

Science.gov (United States)

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.

Bacterial flora of soil after application of oily waste

Energy Technology Data Exchange (ETDEWEB)

Jensen, V

1975-01-01

The influence of mineral oils and oily waste on the bacterial flora of soil was studied both in the field and in model experiments by plate counts followed by examination of the composition of the bacterial flora developing on the plates and by enrichment cultures followed by isolation of pure cultures. A strong increase in bacterial numbers after oil application was observed both in field and model experiments, and this increase occurred within all groups of bacteria, except spore formers and streptomycetes. The most important species of oil decomposing bacteria belonged to the genera Arthrobacter and Pseudomonas.

Effect of alginate chemical disinfection on bacterial count over gypsum cast.

Science.gov (United States)

Haralur, Satheesh B; Al-Dowah, Omir S; Gana, Naif S; Al-Hytham, Abdullah

2012-05-01

To evaluate the efficacy of sodium hypochlorite (1 : 10) and iodophor disinfectants on alginate impressions along with their effect on the survived bacterium count on the gypsum cast. Four alginate impression on each dentate patients were made, of which Group I were not washed or disinfected, Group II impressions were merely washed with water, Group III were disinfected by spraying with sodium hypochlorite (1 : 10), Group IV were disinfected with iodophor (1 : 213). Gypsum cast (type III) were made from all the impression. Impressions and gypsum cast were swabbed in mid palatal region for bacterial culture. Bacterial colony counting done after 3 days of incubation at 37℃ in blood agar media. The data obtained was analyzed by one way ANOVA test at a significant difference level of 0.05. Group I and Group II showed significantly more bacteria compared to Group III and Group IV. Bacterial colonies on the alginate impression and gypsum cast in group disinfected with Sodium hypochlorite (1 : 10) were 0.18, 0.82 respectively compared to group treated with iodophor (1 : 213). There was an increase in bacterial count on dental cast compared to source alginate impressions. Sodium hypochlorite (1 : 10) was found to be better disinfectant for alginate impression. There was an indication of increase in number of bacteria from alginate impression to making of dental cast. Additional gypsum cast disinfectant procedures need to be encouraged to completely eliminate cross infection to dental laboratory.

Using Fluorescence Intensity of Enhanced Green Fluorescent Protein to Quantify Pseudomonas aeruginosa

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Erin Wilson

2018-05-01

Full Text Available A variety of direct and indirect methods have been used to quantify planktonic and biofilm bacterial cells. Direct counting methods to determine the total number of cells include plate counts, microscopic cell counts, Coulter cell counting, flow cytometry, and fluorescence microscopy. However, indirect methods are often used to supplement direct cell counting, as they are often more convenient, less time-consuming, and require less material, while providing a number that can be related to the direct cell count. Herein, an indirect method is presented that uses fluorescence emission intensity as a proxy marker for studying bacterial accumulation. A clinical strain of Pseudomonas aeruginosa was genetically modified to express a green fluorescent protein (PA14/EGFP. The fluorescence intensity of EGFP in live cells was used as an indirect measure of live cell density, and was compared with the traditional cell counting methods of optical density (OD600 and plate counting (colony-forming units (CFUs. While both OD600 and CFUs are well-established methods, the use of fluorescence spectroscopy to quantify bacteria is less common. This study demonstrates that EGFP intensity is a convenient reporter for bacterial quantification. In addition, we demonstrate the potential for fluorescence spectroscopy to be used to measure the quantity of PA14/EGFP biofilms, which have important human health implications due to their antimicrobial resistance. Therefore, fluorescence spectroscopy could serve as an alternative or complementary quick assay to quantify bacteria in planktonic cultures and biofilms.

A novel, optical, on-line bacteria sensor for monitoring drinking water quality

DEFF Research Database (Denmark)

Højris, Bo; Christensen, Sarah Christine Boesgaard; Albrechtsen, Hans-Jørgen

2016-01-01

Today, microbial drinking water quality is monitored through either time-consuming laboratory methods or indirect on-line measurements. Results are thus either delayed or insufficient to support proactive action. A novel, optical, on-line bacteria sensor with a 10-minute time resolution has been...... and quantifying bacteria and particles in pure and mixed suspensions, and the quantification correlates with total bacterial counts. Several field applications have demonstrated that the technology can monitor changes in the concentration of bacteria, and is thus well suited for rapid detection of critical...... conditions such as pollution events in drinking water....

Irradiation control of pathogenic bacteria and their growth during storage in short neckde clam

International Nuclear Information System (INIS)

Ju Hua; Wang Feng; Ha Yiming; Liu Shuliang

2009-01-01

The growth of pathogenic bacteria during storage and their D 10 values by irradiation in short necked clam were evaluated. The results showed that the D 10 values of the total bacteria count, E. coli, Vibrio parahaemolyticus in short necked clam at 0 degree C ∼ 4 degree C condition were 0.70, 0.56, 0.20kGy, respectively. The results showed that total bacteria count in vacuum packaged short necked clam sample was 4.46 log (CFU/g) after 3kGy 60 Co γ-irradiation and 28 days stored under 0 degree C ∼ 4 degree C, which could meet the requirement of related aquatic product standards, the E. coli and Vibrio parahaemolyticus would not detected after 0.4kGy irradiation and 7 days storage at 0 degree C ∼ 4 degree C. Therefore, micro-organisms could be controled effectively by 3 kGy irradiation and its shelf-life could extend to 28 days. (authors)

Prevalence of Vibrio vulnificus and Vibrio parahaemolyticus in the Maryland Coastal Bays

Science.gov (United States)

De Pascuale, V. O.

2016-02-01

The bacterial family of Vibrionaceae is indigenous in the marine estuarine environments such as the Maryland Coastal Bays. Vibrio vulnificus and Vibrio parahaemolyticus are both pathogenic bacteria. Understanding the distribution of Vibrio species is crucial because of the health concerns associated with the bacteria. The aim of this study was to evaluate the overall abundance of bacteria with a focus on Vibrio species in the Maryland Coastal Bays. Seawater samples were collected from 10 different sites that differ with regard to water quality. The total bacteria count (TBC) was determined by two methods: Total plate count and Epifluorescence microscopy. The most-probable-number (MPN) methodology was used to estimate the population of Vibrio parahaemolyticus and Vibrio vulnificus. In addition to the bacteriological analysis, the environmental parameters of temperature and salinity were measured using YSI 6600 multiparameter meter. The average total bacteria count was 2.21 log CFU ml-1. Vibrio vulnificus comprised 5% of the total bacteria count while Vibrio parahaemolyticus comprised only 2% of the total bacteria count. Vibrio vulnificus ranged from 0.30 to 2.48 log MPN ml-1 at the sites tested. Lower Vibrio parahaemolyticus count was observed at the sites with a range of 0.30 to 1.97 log MPN ml-1. There was no significant correlation between the environmental parameters and the Vibrio spp. Since both Vibrio vulnificus and Vibrio parahaemolyticus peak in the summer, there is a potential for a risk of wound infections and gastrointestinal illness based on this data.

A Method for Quantification of Epithelium Colonization Capacity by Pathogenic Bacteria

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Rune M. Pedersen

2018-02-01

Full Text Available Most bacterial infections initiate at the mucosal epithelium lining the gastrointestinal, respiratory, and urogenital tracts. At these sites, bacterial pathogens must adhere and increase in numbers to effectively breach the outer barrier and invade the host. If the bacterium succeeds in reaching the bloodstream, effective dissemination again requires that bacteria in the blood, reestablish contact to distant endothelium sites and form secondary site foci. The infectious potential of bacteria is therefore closely linked to their ability to adhere to, colonize, and invade epithelial and endothelial surfaces. Measurement of bacterial adhesion to epithelial cells is therefore standard procedure in studies of bacterial virulence. Traditionally, such measurements have been conducted with microtiter plate cell cultures to which bacteria are added, followed by washing procedures and final quantification of retained bacteria by agar plating. This approach is fast and straightforward, but yields only a rough estimate of the adhesive properties of the bacteria upon contact, and little information on the ability of the bacterium to colonize these surfaces under relevant physiological conditions. Here, we present a method in which epithelia/endothelia are simulated by flow chamber-grown human cell layers, and infection is induced by seeding of pathogenic bacteria on these surfaces under conditions that simulate the physiological microenvironment. Quantification of bacterial adhesion and colonization of the cell layers is then performed by in situ time-lapse fluorescence microscopy and automatic detection of bacterial surface coverage. The method is demonstrated in three different infection models, simulating Staphylococcus aureus endothelial infection and Escherichia coli intestinal- and uroepithelial infection. The approach yields valuable information on the fitness of the bacterium to successfully adhere to and colonize epithelial surfaces and can be used

Bacterial spot and early blight biocontrol by epiphytic bacteria in tomato plants

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Roberto Lanna Filho

2010-12-01

Full Text Available The objective of this work was to evaluate in vitro and in vivo biocontrol of bacterial spot (Xanthomonas vesicatoria and early blight (Alternaria solani by the epiphytic bacteria Paenibacillus macerans and Bacillus pumilus. Tomato plants were previously sprayed with epiphytic bacteria, benzalkonium chloride and PBS buffer and, after four days, they were inoculated with A. solani and X. vesicatoria. To determine the phytopathogenic bacteria population, leaflet samples were collected from each treatment every 24 hours, for seven days, and plated on semi-selective medium. The effect of epiphytic bacteria over phytopathogens was performed by the antibiosis test and antagonistic activity measured by inhibition zone diameter. The epiphytic and benzalkonium chloride drastically reduced the severity of early blight and bacterial spot in comparison to the control (PBS. In detached leaflets, the epiphytic bacteria reduced in 70% the number of phytopathogenic bacteria cells in the phylloplane. The antibiosis test showed that the epiphytic bacteria efficiently inhibit the phytopathogens growth. In all the bioassays, the epiphytic bacteria protect tomato plants against the phytopathogens

Microbial assessment of un-bottled synthetic juices sold in Peshawar

International Nuclear Information System (INIS)

Abid, H.; Ali, J.; Hussain, A.

2010-01-01

Un-bottled synthetic juices are sold by street vendors and hawkers raising the concern about their safety. To examine the quality of un-bottled synthetic beverages from different locations of Peshawar city using Standard techniques. A total of 56 samples of un-bottled synthetic juices were collected from 8 different locations of Peshawar City and analyzed for Total Plate Count, Total Coliform Bacteria, Total Fecal Coliform bacteria E. coli, Yeast and Mould. In all localities, the street vended un-bottled synthetic juices were found hygienically of poor quality as all had high total plate count which ranged from 2 x 102 to 5 x 107 and total coliform bacteria ranged from 110 MPN/ml. Total Fecal Coliform ranged from < 0.3 to 110M PN/ml. The presence of E coli, contamination was found in (25) 44 % Samples, while all the analyzed samples were contaminated with yeasts and moulds. All un-bottled synthetic juices sold on roadside were highly contaminated with disease causing microorganisms. Policy message: Periodic monitoring of street beverage should be carried out to make them safe for consumption. (author)

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

Science.gov (United States)

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.

Determination of total bacterial count in raw milk by flow cytometry

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Dubravka Samaržija

2004-01-01

Full Text Available The automatic flow cytometry as routine method for total bacterial count determination of raw ex-farm milk has recently been accepted in Croatia. This method significantly differs from the reference method (Standard Plate Count mostly in the presentation of the results obtained. Therefore, this paper summarized experiences in the application of flow cytometry in the dairy laboratories practice. The principle and the practice of the method, methodological details and factors influencing the results were described. In order to avoid problems regarding the interpretation of the results, which aregeneral problems of the quantitative microbiology, this article try to explain an appropriate conversion of the results with regards to SPC/ml, as an official method for the bacteriological quality proposal by the national legislation.

Investigations of the effect of electron-beam irradiation on bacteria in sewage sludge

International Nuclear Information System (INIS)

Osterstock, G.

1976-01-01

The effect of electron beams on bacteria was investigated in 2 experimental facilities. A 400 kV electron accelerator was used to irradiate sludge quantities of 10 l and 180 l. The total bacterial count, the number of coliform bacteria and, in injected sludge, the relative bacteria density of salmonella were investigated. A dose of 0.5 to 0.75 Mrad was required to reduce coliform bacteria to below the detectable level in 0.1 ml. With a dose of 1.5 Mrad salmonella were reduced by 6 orders of magnitude on the average. In addition, the dependence of the reduction in bacteria on the dose rate as well as on mixing of the irradiated material was investigated. Substantial reproduction of bacteria in digested sludge was found in all cases after the irradiation. (orig./MG) [de

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

Science.gov (United States)

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 bacterial cultures in the nonhealing context. Practitioner social (p 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.

Viable bacteria associated with red blood cells and plasma in freshly drawn blood donations.

Science.gov (United States)

Damgaard, Christian; Magnussen, Karin; Enevold, Christian; Nilsson, Martin; Tolker-Nielsen, Tim; Holmstrup, Palle; Nielsen, Claus Henrik

2015-01-01

Infection remains a leading cause of post-transfusion mortality and morbidity. Bacterial contamination is, however, detected in less than 0.1% of blood units tested. The aim of the study was to identify viable bacteria in standard blood-pack units, with particular focus on bacteria from the oral cavity, and to determine the distribution of bacteria revealed in plasma and in the red blood cell (RBC)-fraction. Cross-sectional study. Blood were separated into plasma and RBC-suspensions, which were incubated anaerobically or aerobically for 7 days on trypticase soy blood agar (TSA) or blue lactose plates. For identification colony PCR was performed using primers targeting 16S rDNA. Blood donors attending Capital Region Blood Bank, Copenhagen University Hospital, Rigshospitalet, Hvidovre, Denmark, October 29th to December 10th 2013. 60 donors (≥50 years old), self-reported medically healthy. Bacterial growth was observed on plates inoculated with plasma or RBCs from 62% of the blood donations. Growth was evident in 21 (35%) of 60 RBC-fractions and in 32 (53%) of 60 plasma-fractions versus 8 of 60 negative controls (p = 0.005 and p = 2.6x10-6, respectively). Propionibacterium acnes was found in 23% of the donations, and Staphylococcus epidermidis in 38%. The majority of bacteria identified in the present study were either facultative anaerobic (59.5%) or anaerobic (27.8%) species, which are not likely to be detected during current routine screening. Viable bacteria are present in blood from donors self-reported as medically healthy, indicating that conventional test systems employed by blood banks insufficiently detect bacteria in plasma. Further investigation is needed to determine whether routine testing for anaerobic bacteria and testing of RBC-fractions for adherent bacteria should be recommended.

Monitoring of drug resistance amplification and attenuation with the use of tetracycline-resistant bacteria during wastewater treatment

Science.gov (United States)

Harnisz, Monika; Korzeniewska, Ewa; Niestępski, Sebastian; Osińska, Adriana; Nalepa, Beata

2017-11-01

The objective of this study was to monitor changes (amplification or attenuation) in antibiotic resistance during wastewater treatment based on the ecology of tetracycline-resistant bacteria. The untreated and treated wastewater were collected in four seasons. Number of tetracycline-(TETR) and oxytetracycline-resistant (OTCR) bacteria, their qualitative composition, minimum inhibitory concentrations (MICs), sensitivity to other antibiotics, and the presence of tet (A, B, C, D, E) resistance genes were determined. TETR and OTCR counts in untreated wastewater were 100 to 1000 higher than in treated effluent. OTCR bacterial counts were higher than TETR populations in both untreated and treated wastewater. TETR isolates were not dominated by a single bacterial genus or species, whereas Aeromonas hydrophila and Aeromonas sobria were the most common in OTCR isolates. The treatment process attenuated the drug resistance of TETR bacteria and amplified the resistance of OTCR bacteria. In both microbial groups, the frequency of tet(A) gene increased in effluent in comparison with untreated wastewater. Our results also indicate that treated wastewater is a reservoir of multiple drug-resistant bacteria as well as resistance determinants which may pose a health hazard for humans and animals when released to the natural environment.

Azoreductase activity of anaerobic bacteria isolated from human intestinal microflora.

Science.gov (United States)

Rafii, F; Franklin, W; Cerniglia, C E

1990-01-01

A plate assay was developed for the detection of anaerobic bacteria that produce azoreductases. With this plate assay, 10 strains of anaerobic bacteria capable of reducing azo dyes were isolated from human feces and identified as Eubacterium hadrum (2 strains), Eubacterium spp. (2 species), Clostridium clostridiiforme, a Butyrivibrio sp., a Bacteroides sp., Clostridium paraputrificum, Clostridium nexile, and a Clostridium sp. The average rate of reduction of Direct Blue 15 dye (a dimethoxybenzidine-based dye) in these strains ranged from 16 to 135 nmol of dye per min per mg of protein. The enzymes were inactivated by oxygen. In seven isolates, a flavin compound (riboflavin, flavin adenine dinucleotide, or flavin mononucleotide) was required for azoreductase activity. In the other three isolates and in Clostridium perfringens, no added flavin was required for activity. Nondenaturing polyacrylamide gel electrophoresis showed that each bacterium expressed only one azoreductase isozyme. At least three types of azoreductase enzyme were produced by the different isolates. All of the azoreductases were produced constitutively and released extracellularly. Images PMID:2202258

Lactic Acid and Probiotic Bacteria from Fermented and Probiotic Dairy Products

Directory of Open Access Journals (Sweden)

B.K.L. Karna

2007-12-01

Full Text Available Lactic acid and probiotic bacteria were enumerated and isolated from commercially available yoghurt andprobiotic milk products. Lactobacillus delbrueckii ssp. bulgaricus were enumerated and isolated usingMRS agar incubated anaerobically at 37oC for 72 hrs. M17 agar was used for the enumeration andisolation of Streptococcus thermophilus incubated aerobically at 37oC for 48 hrs. MRS agar and modifiedMRS agar (MRS + L-cysteine + LiCl + Na propionate were used for the enumeration and isolation ofprobiotic bacteria. Both were incubated anaerobically at 37oC for 72 hrs. Morphological, physiologicaland biochemical reactions were used to characterize the isolates.Str. thermophilus counts ranged from 2.6 x 1011 to 2.9 x 1020 CFU/g with Fruit Yoghurt (FY having thehighest count and Yoghurt Natural (YN with the lowest count. Highest Lactobacillus delbrueckii ssp.bulgaricus count was obtained in Duo Yoghurt (DY, 1.1 x 109 and lowest in Yoghurt Drink (YD, 8.0 x 107CFU/g. The highest probiotic bacterial count of 2.3 x 108 was obtained in Yakult (YK and Neslac (Nesshowed the lowest, 1.6 x 102 CFU/g. The viable counts of all the products examined met the prescribedminimum viable count of 105 to 106CFU/g for the claimed health benefits for the consumer except forChamyto Plain (CP, Nes and Nan-2 (Nan.Morphological, physiological and biochemical characteristics showed that the following genera andspecies were present Pediococcus acidilactici (YN, P. pentosaceus (FY, Lactobacillus delbrueckiidelbrueckii and L. brevis in Non Fat High Calcium Yoghurt (NC, L. acidophilus and L. delbrueckiidelbrueckii (DY, YD, P. damnosus and P. pentosaceus in Chamyto Orange (CO, L. delbrueckii bulgaricus,L. acidophilus, and L. delbrueckii delbrueckii (CP, L. para. paracasei (YK and Bifidobacterium ssp.(Nes and Nan.Of the 28 isolates characterized in this study, 15 were Lactobacillus (5 species, 5 werePediococcus (3 species, 6 were Bifidobacterium (species not identified, and 2 were

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

Science.gov (United States)

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

Teknik Pengawetan Fillet Ikan Nila Merah dengan Senyawa Anti Bakteri asal Lactobacillus Acidophilus dan Bifido Bacteria Biffidum

Directory of Open Access Journals (Sweden)

Dede Saputra

2014-12-01

Full Text Available Red tilapia is a good commodity to be developed because it has a high nutritional value composition, with a protein content 17.8%, fat 2.8%, and others composition. The fillet of red tilapia fish is easy to spoil, because of S. aureus, Salmonella sp., and other microbes. Many methods are used to save and preserve the quality of fillet, such fillet preparation through good sanitation practices, cooling process, but the effort were not optimal. The objectives of this study were to 1 evaluate the potency of antibacterial produced by Lactobacillus acidophilus and Bifidobacteria biffidum to inhibit the growth of spoilage bacteria that contaminated the red tilapia fillet; 2 evaluate the effect of antibacterial compounds produced by Lactobacillus acidophilus and Bifidobacteria biffidum of inhibiting the setback fillet quality, 3 determine the shelf life of red tilapia fillet at room temperature. Antibacterial activity test is done by using the well diffusion method; the rate of deterioration of quality of fish tests done by observing the organoleptic parameters, pH measurement test, total volatile base method. Total number of bacteria were performed by Standard Plate Count (SPC test. The LAB’s are able to inhibit the growth of spoilage bacteria Pseudomonas aeruginosa about 8.67-9.00 mm and Listeria monocytogenes about 8.33-9.00 mm through the well diffusion method. pH values about 5.71-5.74, TVB values about 1,26-21.43 with SPC test about 1.39-4.83 CFU/mL. The antibacterial compounds could inhibit  the rate of deterioration of quality red tilapia fillets until 14 hours.

Bactericidal activity of bio-synthesized silver nanoparticles against human pathogenic bacteria

International Nuclear Information System (INIS)

Abalkhil, Tarad Abdulaziz; Alharbi, Sulaiman Ali; Salmen, Saleh Hussein; Wainwright, Milton

2017-01-01

Green synthesis is an attractive and eco-friendly approach to generate potent antibacterial silver nanoparticles (Ag-NPs). Such particles have long been used to fight bacteria and represent a promising tool to overcome the emergence of antibiotic-resistant bacteria. In this study, green synthesis of Ag-NPs was attempted using plant extracts of Aloe vera, Portulaca oleracea and Cynodon dactylon. The identity and size of Ag-NPs was characterized by ultraviolet–visible spectrophotometer and scanning electron microscopy. Monodispersed Ag-NPs were produced with a range of different sizes based on the plant extract used. The bactericidal activity of Ag-NPs against a number of human pathogenic bacteria was determined using the disc diffusion method. The results showed that Gram positive bacteria were more susceptible than Gram negative ones to these antibacterial agents. The minimum inhibitory concentration was determined using the 96- well plate method. Finally, the mechanism by which Ag-NPs affect bacteria was investigated by SEM analysis. Bacteria treated with Ag-NPs were seen to undergo shrinkage and to lose their viability. This study provides evidence for a cheap and effective method for synthesizing potent bactericidal Ag-NPs and demonstrates their effectiveness against human pathogenic bacteria

An improved method of microencapsulation of probiotic bacteria for their stability in acidic and bile conditions during storage.

Science.gov (United States)

Ding, W K; Shah, N P

2009-03-01

The purpose of this study was to develop a method for applying an extra coating of palm oil and poly-L-lysine (POPL) to alginate (ALG) microcapsules to enhance the survival of probiotic bacteria. Eight strains of probiotic bacteria including Lactobacillus rhamnosus, Bifidobacterium longum, L. salivarius, L. plantarum, L. acidophilus, L. paracasei, B. lactis type Bl-O4, and B. lactis type Bi-07 were encapsulated using alginate alone or alginate with POPL. Electron microscopy was used to measure the size of the microcapsules and to determine their surface texture. To assess if the addition of POPL improved the viability of probiotic bacteria in acidic conditions, both ALG and POPL microcapsules were inoculated into pH 2.0 MRS broths and their viability was assessed over a 2-h incubation period. Two bile salts including oxgall bile salt and taurocholic acid were used to test the bile tolerance of probiotic bacteria entrapped in ALG and POPL microcapsules. To assess the porosity and the ability of the microcapsule to hold small molecules in an aqueous environment a water-soluble fluorescent dye, 6-carboxyflourescin (6 FAM), was encapsulated and its release was monitored using a UV spectrophotometer. The results indicated that coating the microcapsules with POPL increased the overall size of the capsules by an average of 3 microm +/- 0.67. However, microcapsules with added POPL had a much smoother surface texture when examined under an electron microscope. The results also indicated that the addition of POPL to microcapsules improved the average viability of probiotic bacteria by > 1 log CFU/mL when compared to ALG microcapsules at 2 h of exposure to acidic conditions. However, similar plate counts were observed between ALG and POPL microcapsules when exposed to bile salts. This suggests that an extra coating of POPL could be readily broken down by bile salts that are commonly found in the lower gastrointestinal tract (GIT). Upon testing the porosity of the

Diversity of root nodule bacteria from leguminous crops

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Agrawal Pooja

2016-01-01

Full Text Available In the present study, a total of 353 nodule-associated bacteria were isolated from 220 legume plant samples belonging to Cicer arietinum (85, Glycine max (74, Vigna radiata (21 and Cajanus cajan (40. A total of 224 bacteria were identified as fast-growing Rhizobium spp. on the basis of differential staining (Gram staining and carbol fuchsin staining and biochemical tests. All the isolates were tested for indole acetic acid production (IAA, phosphate solubilization and siderophore production on plate assay. To examine the effect of volatile organic metabolites (VOM and water soluble soil components (WSSC on nodule bacteria, culture conditions were optimized by observing the effects of various parameters such as pH, salt content and temperatures on the growth of bacteria. Selected rhizobia were subjected to random amplified polymorphic DNA (RAPD and amplified ribosomal DNA restriction analysis (ARDRA analysis to identify their species. On the basis of RAPD and ARDRA, 10 isolates were identified as Rhizobium meliloti. In this study, Rhizobium GO4, G16, G20, G77, S43, S81, M07, M37, A15 and A55 were observed as the best candidates among the tested bacteria and can be further used as potent bioinoculants.

Viable Bacteria Associated with Red Blood Cells and Plasma in Freshly Drawn Blood Donations

DEFF Research Database (Denmark)

Damgaard, Christian; Magnussen, Karin; Enevold, Christian

2015-01-01

) or blue lactose plates. For identification colony PCR was performed using primers targeting 16S rDNA. SETTING: Blood donors attending Capital Region Blood Bank, Copenhagen University Hospital, Rigshospitalet, Hvidovre, Denmark, October 29th to December 10th 2013. PARTICIPANTS: 60 donors (≥50 years old....... CONCLUSIONS: Viable bacteria are present in blood from donors self-reported as medically healthy, indicating that conventional test systems employed by blood banks insufficiently detect bacteria in plasma. Further investigation is needed to determine whether routine testing for anaerobic bacteria and testing......OBJECTIVES: Infection remains a leading cause of post-transfusion mortality and morbidity. Bacterial contamination is, however, detected in less than 0.1% of blood units tested. The aim of the study was to identify viable bacteria in standard blood-pack units, with particular focus on bacteria from...

Investigation of the Rate of the Bacterial Contamination of the Ice Factories in Bandar Abbas, Iran

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Nahid Moradi

2016-09-01

Full Text Available Background: Pollution of drinking water and ice is one of the most serious ways of water borne diseases spread. The purpose of this study was to investigate the bacterial contamination of the ice produced by ice factories in Bandar Abbas. Methods: In this descriptive ,cross-sectional study samples were collected from seven ice factories in Bandar Abbas. Sampling was done by standard method. Amount of the Contamination of ice and water to coliforms was investigated by the Multiple Tube method to determine the MPN, isolate bacteria and identify the microorganisms using conventional bacteriological techniques and counting the total count of bacteria by the Plate Count method on a nutrient agar medium. The data was analyzed by the SPSS software. Results: In this study, a total of 84 samples were investigated. Gram-positive bacteria (65.5% and gram-negative bacteria(34.5% were separated from each other. The MPN rate in samples and total count of bacteria were 0->1100 and 2×101×104 CFU/mL, respectively. Conclusion: The results suggest that necessary precautions be taken by environmental health specialists and other public health authorities in production, transportation and distribution of the ice blocks to reduce the rate of bacterial contamination.

In vitro screening of soil bacteria for inhibiting phytopathogenic fungi ...

African Journals Online (AJOL)

At present, the greatest interest resides with the development and application of specific biocontrol agent for the control of diseases on plant and this form the focus of this work. Several soil bacteria were evaluated in vitro for their effectiveness on the basis of their ability to suppress fungi in plate inhibition assays. 51 strains ...

Fluorescence techniques to detect and to assess viability of plant pathogenic bacteria

NARCIS (Netherlands)

Chitarra, L.G.

2001-01-01

Plant pathogenic bacteria cause major economic losses in commercial crop production worldwide every year. The current methods used to detect and to assess the viability of bacterial pathogens and to test seed lots or plants for contamination are usually based on plate assays or on

A colorimetric assay of 1-aminocyclopropane-1-carboxylate (ACC) based on ninhydrin reaction for rapid screening of bacteria containing ACC deaminase.

Science.gov (United States)

Li, Z; Chang, S; Lin, L; Li, Y; An, Q

2011-08-01

1-Aminocyclopropane-1-carboxylate (ACC) deaminase activity is an efficient marker for bacteria to promote plant growth by lowering ethylene levels in plants. We aim to develop a method for rapidly screening bacteria containing ACC deaminase, based on a colorimetric ninhydrin assay of ACC. A reliable colorimetric ninhydrin assay was developed to quantify ACC using heat-resistant polypropylene chimney-top 96-well PCR plates, having the wells evenly heated in boiling water, preventing accidental contamination from boiling water and limiting evaporation. With this method to measure bacterial consumption of ACC, 44 ACC-utilizing bacterial isolates were rapidly screened out from 311 bacterial isolates that were able to grow on minimal media containing ACC as the sole nitrogen source. The 44 ACC-utilizing bacterial isolates showed ACC deaminase activities and belonged to the genus Burkholderia, Pseudomonas or Herbaspirillum. Determination of bacterial ACC consumption by the PCR-plate ninhydrin-ACC assay is a rapid and efficient method for screening bacteria containing ACC deaminase from a large number of bacterial isolates. The PCR-plate ninhydrin-ACC assay extends the utility of the ninhydrin reaction and enables a rapid screening of bacteria containing ACC deaminase from large numbers of bacterial isolates. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

Fitting a distribution to microbial counts: Making sense of zeroes

DEFF Research Database (Denmark)

Ribeiro Duarte, Ana Sofia; Stockmarr, Anders; Nauta, Maarten

2015-01-01

The accurate estimation of true prevalence and concentration of microorganisms in foods is an important element of quantitative microbiological risk assessment (QMRA). This estimation is often based on microbial detection and enumeration data. Among such data are artificial zero counts, that orig......The accurate estimation of true prevalence and concentration of microorganisms in foods is an important element of quantitative microbiological risk assessment (QMRA). This estimation is often based on microbial detection and enumeration data. Among such data are artificial zero counts......, that originated by chance from contaminated food products. When these products are not differentiated from uncontaminated products that originate true zero counts, the estimates of true prevalence and concentration may be inaccurate. This inaccuracy is especially relevant in situations where highly pathogenic...... bacteria are involved and where growth can occur along the food pathway. Our aim was to develop a method that provides accurate estimates of concentration parameters and differentiates between artificial and true zeroes, thus also accurately estimating true prevalence. We first show the disadvantages...

Modeling particulate removal in plate-plate and wire-plate electrostatic precipitators

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S Ramechecandane

2016-09-01

Full Text Available The present study is concerned with the modeling of electrically charged particles in a model plate-plate and a single wire-plate electrostatic precipitator (ESP. The particle concentration distributions for both a plate-plate and a wire-plate ESP are calculated using a modified drift flux model. Numerical investigations are performed using the modified drift flux model for particle number concentration, in addition to the RNG k - ε model for the mean turbulent flow field and the Poisson equation for the electric field. The proposed model and the outlined methodology for coupling the flow field, electric field, charging kinetics and particle concentration is applied to two model precipitators that are truly representative of a wide class of commercialized ESPs. The present investigation is quite different from the earlier studies as it does not make assumptions like a homogeneous electric field or an infinite turbulent diffusivity. The electric field calculated is a strong function of position and controls the migration velocity of particles. Hence, the proposed model can be implemented in a flow solver to obtain a full-fledged solution for any kind of ESP with no limitations on the particle number concentration, as encountered in a Lagrangian approach. The effect of turbulent diffusivity on particle number concentration in a plate-plate ESP is investigated in detail and the results obtained are compared with available experimental data. Similarly, the effect of particle size/diameter and applied electric potential on the accumulative collection performance in the case of a wire-plate ESP is studied and the results obtained are compared with available numerical data. The numerical results obtained using the modified drift flux model for both the plate-plate and wire-plate ESP are in close agreement with available experimental and numerical data.

Predictive models for the effect of storage temperature on Vibrio parahaemolyticus viability and counts of total viable bacteria in Pacific oysters (Crassostrea gigas).

Science.gov (United States)

Fernandez-Piquer, Judith; Bowman, John P; Ross, Tom; Tamplin, Mark L

2011-12-01

Vibrio parahaemolyticus is an indigenous bacterium of marine environments. It accumulates in oysters and may reach levels that cause human illness when postharvest temperatures are not properly controlled and oysters are consumed raw or undercooked. Predictive models were produced by injecting Pacific oysters (Crassostrea gigas) with a cocktail of V. parahaemolyticus strains, measuring viability rates at storage temperatures from 3.6 to 30.4°C, and fitting the data to a model to obtain parameter estimates. The models were evaluated with Pacific and Sydney Rock oysters (Saccostrea glomerata) containing natural populations of V. parahaemolyticus. V. parahaemolyticus viability was measured by direct plating samples on thiosulfate-citrate-bile salts-sucrose (TCBS) agar for injected oysters and by most probable number (MPN)-PCR for oysters containing natural populations. In parallel, total viable bacterial counts (TVC) were measured by direct plating on marine agar. Growth/inactivation rates for V. parahaemolyticus were -0.006, -0.004, -0.005, -0.003, 0.030, 0.075, 0.095, and 0.282 log₁₀ CFU/h at 3.6, 6.2, 9.6, 12.6, 18.4, 20.0, 25.7, and 30.4°C, respectively. The growth rates for TVC were 0.015, 0.023, 0.016, 0.048, 0.055, 0.071, 0.133, and 0.135 log₁₀ CFU/h at 3.6, 6.2, 9.3, 14.9, 18.4, 20.0, 25.7, and 30.4°C, respectively. Square root and Arrhenius-type secondary models were generated for V. parahaemolyticus growth and inactivation kinetic data, respectively. A square root model was produced for TVC growth. Evaluation studies showed that predictive growth for V. parahaemolyticus and TVC were "fail safe." The models can assist oyster companies and regulators in implementing management strategies to minimize V. parahaemolyticus risk and enhancing product quality in supply chains.

Decontamination effect of milling by a jet mill on bacteria in rice flour.

Science.gov (United States)

Sotome, Itaru; Nei, Daisuke; Tsuda, Masuko; Mohammed, Sharif Hossen; Takenaka, Makiko; Okadome, Hiroshi; Isobe, Seiichiro

2011-06-01

The decontamination effect of milling by a jet mill was investigated by counting the number of bacteria in brown and white rice flour with mean particle diameters of 3, 20, and 40µm prepared by the jet mill. In the jet mill, the particles are crushed and reduced in size by the mechanical impact caused by their collision. Although the brown and white rice grains were contaminated with approximately 10(6) and 10(5) CFU/g bacteria, the microbial load of the rice flour decreased as the mean particle diameter decreased, ultimately decreasing to approximately 104 and 103 CFU/g in the brown and white rice flour. The temperature and pressure changes of the sample were not considered to have an effect on reducing the bacterial count during the milling. Hence, it was thought that the rice flour was decontaminated by other effects.

Development of a Flow Cytometry-Based Method for Rapid Detection of Escherichia coli and Shigella Spp. Using an Oligonucleotide Probe

Science.gov (United States)

Xue, Yong; Wilkes, Jon G.; Moskal, Ted J.; Williams, Anna J.; Cooper, Willie M.; Nayak, Rajesh; Rafii, Fatemeh; Buzatu, Dan A.

2016-01-01

Standard methods to detect Escherichia coli contamination in food use the polymerase chain reaction (PCR) and agar culture plates. These methods require multiple incubation steps and take a long time to results. An improved rapid flow-cytometry based detection method was developed, using a fluorescence-labeled oligonucleotide probe specifically binding a16S rRNA sequence. The method positively detected 51 E. coli isolates as well as 4 Shigella species. All 27 non-E. coli strains tested gave negative results. Comparison of the new genetic assay with a total plate count (TPC) assay and agar plate counting indicated similar sensitivity, agreement between cytometry cell and colony counts. This method can detect a small number of E.coli cells in the presence of large numbers of other bacteria. This method can be used for rapid, economical, and stable detection of E. coli and Shigella contamination in the food industry and other contexts. PMID:26913737

Development of a Flow Cytometry-Based Method for Rapid Detection of Escherichia coli and Shigella Spp. Using an Oligonucleotide Probe.

Directory of Open Access Journals (Sweden)

Yong Xue

Full Text Available Standard methods to detect Escherichia coli contamination in food use the polymerase chain reaction (PCR and agar culture plates. These methods require multiple incubation steps and take a long time to results. An improved rapid flow-cytometry based detection method was developed, using a fluorescence-labeled oligonucleotide probe specifically binding a16S rRNA sequence. The method positively detected 51 E. coli isolates as well as 4 Shigella species. All 27 non-E. coli strains tested gave negative results. Comparison of the new genetic assay with a total plate count (TPC assay and agar plate counting indicated similar sensitivity, agreement between cytometry cell and colony counts. This method can detect a small number of E.coli cells in the presence of large numbers of other bacteria. This method can be used for rapid, economical, and stable detection of E. coli and Shigella contamination in the food industry and other contexts.

Quality aspects of raw goat milk in Lower Southern Thailand

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Siriwat Wasiksiri

2010-05-01

Full Text Available This study was conducted to determine the quality and safety of raw goat milk in Lower Southern Thailand duringAugust to September 2008. Milk samples were collected from five farms in Songkhla, Yala, and Pattani Province of which thepH, acidity, specific gravity, milk fat, solid not fat, total solid, total plate count, Coliform count, and antibiotic residue weretested. The results did not show any significant difference (p>0.05 on pH and total plate count among samples from eachfarm. However, acidity, specific gravity, milk fat, solid not fat, and total solid varied between farms depending on feed supplyand management. Means of total bacteria count and Coliform count of most samples were in TACF standard quality (log3.720 cfu/ml and log 1.892 cfu/ml, respectively, except four samples had higher Coliform contamination. Additionally,a higher proportion (22.7 % of samples with antibiotic residue was found.

A directional gamma-ray detector based on scintillator plates

Energy Technology Data Exchange (ETDEWEB)

Hanna, D., E-mail: hanna@physics.mcgill.ca; Sagnières, L.; Boyle, P.J.; MacLeod, A.M.L.

2015-10-11

A simple device for determining the azimuthal location of a source of gamma radiation, using ideas from astrophysical gamma-ray burst detection, is described. A compact and robust detector built from eight identical modules, each comprising a plate of CsI(Tl) scintillator coupled to a photomultiplier tube, can locate a point source of gamma rays with degree-scale precision by comparing the count rates in the different modules. Sensitivity to uniform environmental background is minimal.

Contamination Levels and Identification of Bacteria in Milk Sampled from Three Regions of Tanzania: Evidence from Literature and Laboratory Analyses

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G. Msalya

2017-01-01

Full Text Available Milk in Tanzania has been reported to be contaminated with large number of bacteria. This is because (1 milk is obtained from animals with unknown health status, (2 good milking and handling practices are to a large extent not observed, and (3 marketing and distribution are done in informal channels. These factors are potential causes of milk-borne diseases and milk quality loss. The aim of this study was to assess nutritional risks in milk as reported in literature over a period of 20 years and through analyses of samples collected during the present study. The issues highlighted in literature were high bacteria and coliform counts exceeding standard levels in East Africa, prevalence of bacteria and drug residues in milk, and adulteration. Based on performed analyses, total bacterial count 1.0×107 colony forming units per millilitre (cfu/ml and total coliform count 1.1×107 cfu/ml, also greater than recommended levels, were found. Ten bacteria types were isolated from milk samples (five, Pseudomonas aeruginosa, Listeria monocytogenes, Listeria innocua, Listeria ivanovii, and Klebsiella spp. are reported in Tanzanian for the first time. Two drugs tetracycline and sulphur were detected. Therefore, it is worth noting that integrated research is needed to evaluate the situation and address these challenges.

Preliminary stochastic model for managing Vibrio parahaemolyticus and total viable bacterial counts in a Pacific oyster (Crassostrea gigas) supply chain.

Science.gov (United States)

Fernandez-Piquer, Judith; Bowman, John P; Ross, Tom; Estrada-Flores, Silvia; Tamplin, Mark L

2013-07-01

Vibrio parahaemolyticus can accumulate and grow in oysters stored without refrigeration, representing a potential food safety risk. High temperatures during oyster storage can lead to an increase in total viable bacteria counts, decreasing product shelf life. Therefore, a predictive tool that allows the estimation of both V. parahaemolyticus populations and total viable bacteria counts in parallel is needed. A stochastic model was developed to quantitatively assess the populations of V. parahaemolyticus and total viable bacteria in Pacific oysters for six different supply chain scenarios. The stochastic model encompassed operations from oyster farms through consumers and was built using risk analysis software. Probabilistic distributions and predictions for the percentage of Pacific oysters containing V. parahaemolyticus and high levels of viable bacteria at the point of consumption were generated for each simulated scenario. This tool can provide valuable information about V. parahaemolyticus exposure and potential control measures and can help oyster companies and regulatory agencies evaluate the impact of product quality and safety during cold chain management. If coupled with suitable monitoring systems, such models could enable preemptive action to be taken to counteract unfavorable supply chain conditions.

Sales of Forestry-Related Specialty License Plates in the Southern United States: A County Level Empirical Analysis

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Shaun M. Tanger

2016-07-01

Full Text Available In recent years, specialty license plates have become an increasingly popular way to raise awareness and show support for a myriad of issues with which the plate is linked. Several states and various organizations that provide forestry education have developed forestry license plates. Vehicle owners can purchase the plates to show their support towards forestry by buying the forestry license plates, which generates revenue for the provider organization. Using county-level data from five states in the Southeastern United States, a statistical model was developed to examine explanatory factors of forestry-based specialty license plate sales in 2014. Using linear count regression modeling, we observed that the significant predictor variables of plate sales were income per capita, population density, the percentage of acres that are forested in the county, acres of forest in the county that are privately owned, percentage of people who are 65 or older, and presence of the forest industry in the county. Plate sales were positively correlated with the presence of the forest industry in the county.

Screening of antagonistic bacteria against the green mold disease (Trichoderma harzianum Rifai of Grey Oyster Mushroom (Pleurotus pulmonarius (Fr. Quel.

Directory of Open Access Journals (Sweden)

Nualsri, C.

2005-01-01

Full Text Available A total of 174 strains of bacteria antagonistic against the green mold (Trichoderma harzianum, isolated from cultivating bags and fruiting bodies of the mushrooms, were screened for effects on mushroom mycelia and ability to control the green mold disease. Twenty-eight of them promoted the primodia formation of the Pleurotus pulmonarius mycelia on agar plates. Twenty-two isolates were selected and further tested in a mushroom house. Cell suspension of each isolate was prepared and sprayed onto the spawn surface of P. pulmonarius. Fifteen isolates shortened the times required from watering to 2nd and 3rd flushing and increased yield of the basidiocarps by 1.1-34.3% over 30 days. Six isolates of bacteria which showed an inhibitory effect against T. harzianum, enhanced primordia formation and increased yield of P. pulmonarius were selected and used for control testing in a cultivation house. The suspension of each isolate was sprayed onto the spawn surface immediately after exposure to the air in the mushroom house, followed by spore suspension of T. harzianum two days later. The number of infected bags was counted at 30 days after inoculation and the cumulative yield was compared after 60 days. The results showed that bacteria isolate B012-022 was highly effective in suppressing the green mold disease.Only 6.7% of the cultivating bags were found to be infected by T. harzianum when bacteria isolate B012-022 was applied. Cumulative yield obtained from 900 g of 94% sawdust + 5% rice bran + 1% Ca(OH2 was 300.0 g/bag after 60 days, 71.1% higher than the bags infected by the green mold and without bacterial spraying. Identification of the six bacterial isolates showed all to be Bacillus spp.

Photodynamic effects of methylene blue-loaded polymeric nanoparticles on dental plaque bacteria.

Science.gov (United States)

Klepac-Ceraj, Vanja; Patel, Niraj; Song, Xiaoqing; Holewa, Colleen; Patel, Chitrang; Kent, Ralph; Amiji, Mansoor M; Soukos, Nikolaos S

2011-09-01

Photodynamic therapy (PDT) is increasingly being explored for treatment of oral infections. Here, we investigate the effect of PDT on human dental plaque bacteria in vitro using methylene blue (MB)-loaded poly(lactic-co-glycolic) (PLGA) nanoparticles with a positive or negative charge and red light at 665 nm. Dental plaque samples were obtained from 14 patients with chronic periodontitis. Suspensions of plaque microorganisms from seven patients were sensitized with anionic, cationic PLGA nanoparticles (50 µg/ml equivalent to MB) or free MB (50 µg/ml) for 20 min followed by exposure to red light for 5 min with a power density of 100 mW/cm2 . Polymicrobial oral biofilms, which were developed on blood agar in 96-well plates from dental plaque inocula obtained from seven patients, were also exposed to PDT as above. Following the treatment, survival fractions were calculated by counting the number of colony-forming units. The cationic MB-loaded nanoparticles exhibited greater bacterial phototoxicity in both planktonic and biofilm phase compared to anionic MB-loaded nanoparticles and free MB, but results were not significantly different (P > 0.05). Cationic MB-loaded PLGA nanoparticles have the potential to be used as carriers of MB for PDT systems. Copyright © 2011 Wiley-Liss, Inc.

Dynamic states of swimming bacteria in a nematic liquid crystal cell with homeotropic alignment

International Nuclear Information System (INIS)

Zhou, Shuang; Harvard University, Cambridge, MA; Tovkach, Oleh; University of Massachusetts, Amherst, MA; Golovaty, Dmitry

2017-01-01

Flagellated bacteria such as Escherichia coli and Bacillus subtilis exhibit effective mechanisms for swimming in fluids and exploring the surrounding environment. In isotropic fluids such as water, the bacteria change swimming direction through the run-and-tumble process. Lyotropic chromonic liquid crystals (LCLCs) have been introduced recently as an anisotropic environment in which the direction of preferred orientation, the director, guides the bacterial trajectories. In this work, we describe the behavior of bacteria B. subtilis in a homeotropic LCLC geometry, in which the director is perpendicular to the bounding plates of a shallow cell. We demonstrate that the bacteria are capable of overcoming the stabilizing elastic forces of the LCLC and swim perpendicularly to the imposed director (and parallel to the bounding plates). The effect is explained by a finite surface anchoring of the director at the bacterial body; the role of surface anchoring is analyzed by numerical simulations of a rod realigning in an otherwise uniform director field. Shear flows produced by a swimming bacterium cause director distortions around its body, as evidenced both by experiments and numerical simulations. These distortions contribute to a repulsive force that keeps the swimming bacterium at a distance of a few micrometers away from the bounding plates. The homeotropic alignment of the director imposes two different scenarios of bacterial tumbling: one with an 180° reversal of the horizontal velocity and the other with the realignment of the bacterium by two consecutive 90° turns. Finally, in the second case, the angle between the bacterial body and the imposed director changes from 90° to 0° and then back to 90°; the new direction of swimming does not correlate with the previous swimming direction.

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

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Claire Bankier

Full Text Available 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.

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

Science.gov (United States)

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.

Influence of Asellus aquaticus on Escherichia coli, Klebsiella pneumoniae, Campylobacter jejuni and naturally occurring heterotrophic bacteria in drinking water.

Science.gov (United States)

Christensen, Sarah C B; Nissen, Erling; Arvin, Erik; Albrechtsen, Hans-Jørgen

2012-10-15

Water lice, Asellus aquaticus (isopoda), frequently occur in drinking water distribution systems where they are a nuisance to consumers and water utilities. Whether they are solely an aesthetic problem or also affect the microbial water quality is a matter of interest. We studied the influence of A. aquaticus on microbial water quality in non-chlorinated drinking water in controlled laboratory experiments. Pure cultures of the indicator organisms Escherichia coli and Klebsiella pneumoniae and the pathogen Campylobacter jejuni as well as naturally occurring heterotrophic drinking water bacteria (measured as heterotrophic plate counts, HPC) were investigated in microcosms at 7 °C, containing non-sterilised drinking water, drinking water sediment and A. aquaticus collected from a non-chlorinated ground water based drinking water supply system. Concentrations of E. coli, K. pneumoniae and C. jejuni decreased over time, following a first order decay with half lives of 5.3, 18.4 and 1.3 days, respectively. A. aquaticus did not affect survival of indicators and pathogens substantially whereas HPC were influenced by presence of dead A. aquaticus. Growth rates increased with an average of 48% for bacteria grown on R-2A agar and an average of 83% for bacteria grown on yeast extract agar when dead A. aquaticus were present compared to no and living A. aquaticus present. A. aquaticus associated E. coli, K. pneumoniae and C. jejuni were measured (up to 25 per living and 500 per dead A. aquaticus) and so were A. aquaticus associated heterotrophic bacteria (>1.8*10(4) CFU per living and >6*10(4) CFU per dead A. aquaticus). A. aquaticus did not serve as an optimised habitat that increased survival of indicators and pathogens, since A. aquaticus associated E. coli, K. pneumoniae and C. jejuni were only measured as long as the bacteria were also present in the water and sediment. Copyright © 2012 Elsevier Ltd. All rights reserved.

Hiding in Fresh Fruits and Vegetables: Opportunistic Pathogens May Cross Geographical Barriers

OpenAIRE

Al-Kharousi, Zahra S.; Guizani, Nejib; Al-Sadi, Abdullah M.; Al-Bulushi, Ismail M.; Shaharoona, Baby

2016-01-01

Different microbial groups of the microbiome of fresh produce can have diverse effects on human health. This study was aimed at identifying some microbial communities of fresh produce by analyzing 105 samples of imported fresh fruits and vegetables originated from different countries in the world including local samples (Oman) for aerobic plate count and the counts of Enterobacteriaceae, Enterococcus, and Staphylococcus aureus. The isolated bacteria were identified by molecular (PCR) and bioc...

Meta-Analysis of Quantification Methods Shows that Archaea and Bacteria Have Similar Abundances in the Subseafloor

Science.gov (United States)

May, Megan K.; Kevorkian, Richard T.; Steen, Andrew D.

2013-01-01

There is no universally accepted method to quantify bacteria and archaea in seawater and marine sediments, and different methods have produced conflicting results with the same samples. To identify best practices, we compiled data from 65 studies, plus our own measurements, in which bacteria and archaea were quantified with fluorescent in situ hybridization (FISH), catalyzed reporter deposition FISH (CARD-FISH), polyribonucleotide FISH, or quantitative PCR (qPCR). To estimate efficiency, we defined “yield” to be the sum of bacteria and archaea counted by these techniques divided by the total number of cells. In seawater, the yield was high (median, 71%) and was similar for FISH, CARD-FISH, and polyribonucleotide FISH. In sediments, only measurements by CARD-FISH in which archaeal cells were permeabilized with proteinase K showed high yields (median, 84%). Therefore, the majority of cells in both environments appear to be alive, since they contain intact ribosomes. In sediments, the sum of bacterial and archaeal 16S rRNA gene qPCR counts was not closely related to cell counts, even after accounting for variations in copy numbers per genome. However, qPCR measurements were precise relative to other qPCR measurements made on the same samples. qPCR is therefore a reliable relative quantification method. Inconsistent results for the relative abundance of bacteria versus archaea in deep subsurface sediments were resolved by the removal of CARD-FISH measurements in which lysozyme was used to permeabilize archaeal cells and qPCR measurements which used ARCH516 as an archaeal primer or TaqMan probe. Data from best-practice methods showed that archaea and bacteria decreased as the depth in seawater and marine sediments increased, although archaea decreased more slowly. PMID:24096423

Ralicon anodes for image photon counting fabricated by electron beam lithography

International Nuclear Information System (INIS)

Burton, W.M.

1982-01-01

The Anger wedge and strip anode event location system developed for microchannel plate image photon detectors at the Space Sciences Laboratory of the University of California, Berkeley, has been extended in the present work by the use of electron beam lithography (EBL). This method of fabrication can be used to produce optical patterns for the subsequent manufacture of anodes by conventional photo-etching methods and has also enabled anodes to be produced directly by EBL microfabrication techniques. Computer-aided design methods have been used to develop several types of RALICON (Readout Anodes of Lithographic Construction) for use in photon counting microchannel plate imaging detectors. These anodes are suitable for linear, two dimensional or radial position measurements and they incorporate novel design features made possible by the EBL fabrication technique which significantly extend their application relative to published wedge-strip anode designs. (author)

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

Science.gov (United States)

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 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 could potentially result in a

Beverages obtained from soda fountain machines in the U.S. contain microorganisms, including coliform bacteria.

Science.gov (United States)

White, Amy S; Godard, Renee D; Belling, Carolyn; Kasza, Victoria; Beach, Rebecca L

2010-01-31

Ninety beverages of three types (sugar sodas, diet sodas and water) were obtained from 20 self-service and 10 personnel-dispensed soda fountains, analyzed for microbial contamination, and evaluated with respect to U.S. drinking water regulations. A follow-up study compared the concentration and composition of microbial populations in 27 beverages collected from 9 soda fountain machines in the morning as well as in the afternoon. Ice dispensed from these machines was also examined for microbial contamination. While none of the ice samples exceeded U.S. drinking water standards, coliform bacteria was detected in 48% of the beverages and 20% had a heterotrophic plate count greater than 500cfu/ml. Statistical analyses revealed no difference in levels of microbial contamination between beverage types or between those dispensed from self-service and personnel-dispensed soda fountains. More than 11% of the beverages analyzed contained Escherichia coli and over 17% contained Chryseobacterium meningosepticum. Other opportunistic pathogenic microorganisms isolated from the beverages included species of Klebsiella, Staphylococcus, Stenotrophomonas, Candida, and Serratia. Most of the identified bacteria showed resistance to one or more of the 11 antibiotics tested. These findings suggest that soda fountain machines may harbor persistent communities of potentially pathogenic microorganisms which may contribute to episodic gastric distress in the general population and could pose a more significant health risk to immunocompromised individuals. These findings have important public health implications and signal the need for regulations enforcing hygienic practices associated with these beverage dispensers. Copyright 2009 Elsevier B.V. All rights reserved.

Psychrotrophic bacteria and their negative effects on milk and dairy products quality

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Šimun Zamberlin

2012-06-01

Full Text Available The characteristics of bacterial populations in raw milk at the time of processing has a significant influence on shelf-life, organoleptic quality, spoilage and yields of raw milk, processed milk as well as on the other dairy products. Unfortunately, cold and extended storage of raw milk, as a common practice in dairy sector today, favour the growth of psychrotrophic bacteria. Therefore, their count in the refrigerated milk is more than the ideal limit of 10 % of the mesophilic count. Psychrotrophic bacteria are generally able to form extracellular or intracellular thermo-resistant enzymes (proteases, lipases, phospolipases which can contribute to milk and dairy products spoilage. In addition, besides exhibiting spoilage features, some species belonging to the psychrotrops are considered as emerging pathogens that carry innate resistance to antibiotics or produce toxins. In sense of quality, psychrotrophic bacteria have become major problem for today’s dairy industry as leading cause in spoilage of cold-storage milk and dairy products. This review article focuses on the impact of psychrotrops on quality problems associated with raw milk as well as on th final dairy products. Means of controlling the dominant psychrotrophic species responsible for undesirable activities in milk and dairy products were also discussed.

Bacteria contributing to behaviour of radiocarbon in sodium acetate

International Nuclear Information System (INIS)

Ishii, N.; Uchida, S.

2011-01-01

An acetate-utilising bacterium was isolated and identified from deionised water that was used for flooding of paddy soils in this study's batch culture experiments. Bacteria in the deionised water samples formed colonies on agar plates containing [1,2- 14 C] sodium acetate, and the autoradiograms showed that all the colonies were positive for 14 C utilisation. Then one of the acetate-utilising bacteria was isolated. The isolate was characterised by phylogenetic analysis, cell morphology, Gram staining and growth at 30 deg. C. Phylogenetic analysis based on 16 S rRNA sequencing showed that the isolate belonged to the genus Burkholderia. The bacterium was gram-negative rods and grew at 30 deg. C under aerobic conditions. Based on these characteristics, the isolate was identified as Burkholderia gladioli. Because B. gladioli is often found in soil, water and the rhizosphere, attention must be paid to the relationships between bacteria and the behaviour of 14 C to for the safety assessment of geological disposal of transuranic waste. (authors)

Changes in the Microbial Composition of Microbrewed Beer during the Process in the Actual Manufacturing Line.

Science.gov (United States)

Kim, S A; Jeon, S H; Kim, N H; Kim, H W; Lee, N Y; Cho, T J; Jung, Y M; Lee, S H; Hwang, I G; Rhee, M S

2015-12-01

This study investigated changes in the microbial composition of microbrewed beer during the manufacturing processes and identified potential microbial hazards, effective critical quality control points, and potential contamination routes. Comprehensive quantitative (aerobic plate count, lactic acid bacteria, fungi, acetic acid bacteria, coliforms, and Bacillus cereus) and qualitative (Escherichia coli and eight foodborne pathogens) microbiological analyses were performed using samples of raw materials (malt and manufacturing water), semiprocessed products (saccharified wort, boiled wort, and samples taken during the fermentation and maturation process), and the final product obtained from three plants. The initial aerobic plate count and lactic acid bacteria counts in malt were 5.2 and 4.3 log CFU/g, respectively. These counts were reduced to undetectable levels by boiling but were present at 2.9 and 0.9 log CFU/ml in the final product. Fungi were initially present at 3.6 log CFU/g, although again, the microbes were eliminated by boiling; however, the level in the final product was 4.6 log CFU/ml. No E. coli or foodborne pathogens (except B. cereus) were detected. B. cereus was detected at all stages, although it was not present in the water or boiled wort (total detection rate ¼ 16.4%). Results suggest that boiling of the wort is an effective microbial control measure, but careful management of raw materials and implementation of effective control measures after boiling are needed to prevent contamination of the product after the boiling step. The results of this study may constitute useful and comprehensive information regarding the microbiological quality of microbrewed beer.

Hydrodynamics of a flexible plate between pitching rigid plates

Science.gov (United States)

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.

Characterization of Bacteria in Nigerian Yogurt as Promising Alternative to Antibiotics in Gastrointestinal Infections.

Science.gov (United States)

Ayeni, Anthony Opeyemi; Ruppitsch, Werner; Ayeni, Funmilola Abidemi

2018-03-14

Gastrointestinal infections are endemic in Nigeria and several factors contribute to their continual survival, including bacterial resistance to commonly used antibiotics. Nigerian yogurts do not include probiotics, and limited information is available about the antimicrobial properties of the fermenters in the yogurt against gastrointestinal pathogens. Therefore, the antimicrobial potentials of bacteria in Nigeria-produced yogurts against intestinal pathogens were investigated in this study. Viable counts of lactic acid bacteria (LAB) in 15 brands of yogurt were enumerated and the bacteria identified by partial sequencing of 16S rRNA gene. Susceptibility of the gastrointestinal pathogens (Salmonella, Shigella and E. coli ) to antibiotics by disc diffusion method, to viable LAB by the agar overlay method, and to the cell-free culture supernatant (CFCS) of the LAB were investigated. Co-culture analysis of LAB and pathogens were also done. Viable counts of 1.5 × 10 11 cfu/ml were observed in some yogurt samples. Two genera were identified: Lactobacillus (70.7%) and Acetobacter (29.3%). The Lactobacillus species reduced multidrug-resistant gastrointestinal pathogens by 4 to 5 log while the zones of inhibition ranged between 11 and 23. The Lactobacillus and Acetobacter strains examined displayed good activities against the multidrug-resistant tested pathogens. This is the first report of antimicrobial activities of acetic acid bacteria isolated from yogurt in Nigeria.

Evaluation of In-Vacuum Imaging Plate Detector for X-Ray Diffraction Microscopy

International Nuclear Information System (INIS)

Nishino, Yoshinori; Takahashi, Yukio; Yamamoto, Masaki; Ishikawa, Tetsuya

2007-01-01

We performed evaluation tests of a newly developed in-vacuum imaging plate (IP) detector for x-ray diffraction microscopy. IP detectors have advantages over direct x-ray detection charge-coupled device (CCD) detectors, which have been commonly used in x-ray diffraction microscopy experiments, in the capabilities for a high photon count and for a wide area. The detector system contains two IPs to make measurement efficient by recording data with the one while reading or erasing the other. We compared speckled diffraction patterns of single particles taken with the IP and a direct x-ray detection CCD. The IP was inferior to the CCD in spatial resolution and in signal-to-noise ratio at a low photon count

Bacteriological investigations of the irradiation of fresh fish using a new classification scheme for bacteria of the fish skin

Energy Technology Data Exchange (ETDEWEB)

Karnop, G [Bundesforschungsanstalt fuer Fischerei, Hamburg (F.R. Germany). Inst. fuer Biochemie und Technologie

1975-04-01

Investigations were made on the effect of irradiation with 36, 72, 108 and 144 kr, carried out once and twice, on the bacterial flora of the skin of red fish on the day of catching and after 9, 16 and 23 days of storage in ice. With an initial total count of the fish of 13,700 bacteria/cm/sup 2/ of skin surface irradiation with 36 kr (108 kr) on the day of catching caused a reduction of the total count to 11% (1.3%) of the nonirradiated fish on the 9th day of storage. Nearly all differences disappeared by the 16th day. A second irradiation with 36 kr and 108 kr on the 9th day reduced the bacterial count on a large scale by which on the 16th day the total count of these fishes was lower than that of the nonirradiated fish on the 9th day. Later on the differentiation disappeared quickly but there were small differences unlike the nonirradiated fish on the 23rd day. The rapid equalization during the last storage period is possibly only typical of the storage in boxes. A scheme for the characterization of types of spoilage bacteria recently established and based on the bacterial attack on leucine, ..beta..-alanine, creatine, creatinine and cystine yielded the following results: The different Pseudomonas types were reduced much more than Achromobacter types. The irradiation effect does not only consist in a reduction of the general total count of bacteria but also in the selective destruction of the most active spoilage bacteria with a very extensive enzymatic pattern which concerns many organic nitrogen compounds in the tissue of fish. By means of a sub-group of Pseudomonas and several maturity stages of the bacterial populations a 7 days delay of the bacterial evolution, caused by the second irradiation with 36 kr, could be observed. The useful effect of irradiation carried out twice with doses about 50 kr was discussed and estimated at a 10 to 12 days delay of the bacterial spoilage.

Bacteriological investigations of the irradiation of fresh fish using a new classification scheme for bacteria of the fish skin

International Nuclear Information System (INIS)

Karnop, G.

1975-01-01

Investigations were made on the effect of irradiation with 36, 72, 108 and 144 kr, carried out once and twice, on the bacterial flora of the skin of red fish on the day of catching and after 9, 16 and 23 days of storage in ice. With an initial total count of the fish of 13,700 bacteria/cm 2 of skin surface irradiation with 36 kr (108 kr) on the day of catching caused a reduction of the total count to 11% (1.3%) of the nonirradiated fish on the 9th day of storage. Nearly all differences disappeared by the 16th day. A second irradiation with 36 kr and 108 kr on the 9th day reduced the bacterial count on a large scale by which on the 16th day the total count of these fishes was lower than that of the nonirradiated fish on the 9th day. Later on the differentiation disappeared quickly but there were small differences unlike the nonirradiated fish on the 23rd day. The rapid equalization during the last storage period is possibly only typical of the storage in boxes. A scheme for the characterization of types of spoilage bacteria recently established and based on the bacterial attack on leucine, β-alanine, creatine, creatinine and cystine yielded the following results: The different Pseudomonas types were reduced much more than Achromobacter types. The irradiation effect does not only consist in a reduction of the general total count of bacteria but also in the selective destruction of the most active spoilage bacteria with a very extensive enzymatic pattern which concerns many organic nitrogen compounds in the tissue of fish. By means of a sub-group of Pseudomonas and several maturity stages of the bacterial populations a 7 days delay of the bacterial evolution, caused by the second irradiation with 36 kr, could be observed. The useful effect of irradiation carried out twice with doses about 50 kr was discussed and estimated at a 10-12 days delay of the bacterial spoilage. (orig./MG) [de

Evaluation of the Influence of Frequency of Milk Collection and Milking Dayshift on the Microbiological Quality of Raw Milk

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Lucía Reguillo

2018-01-01

Full Text Available The aim of this study was to analyze the influence of milk collection frequency (24 h versus 48 h and milking dayshift (morning and evening on total mesophilic aerobic bacteria (MAB and psychrotrophic bacteria (PSY counts in raw milk samples. MAB counts were determined by flow cytometry (BactoScan and PSY counts by the plate counting agar method. An univariate statistical analysis was performed to find out significant differences among the studied factors. Results obtained showed that collecting milk every 24 h was effective in reducing MAB and PSY counts by 32 and 18%, respectively, compared to 48 h milk collection. This positive impact allowed reducing up to 4°C the temperature of the heat treatment in the dairy industry, thus involving energy savings of 22%. Milking during the mornings showed a significant reduction of MAB counts in comparison to milking performed during the evenings (P<0.05. These results are highly useful for the improvement of milk quality through the optimization of collection and milking systems set at primary production.

Microbial counts of food contact surfaces at schools depending on a feeding scheme

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Nthabiseng Nhlapo

2014-11-01

Full Text Available The prominence of disease transmission between individuals in confined environments is a concern, particularly in the educational environment. With respect to school feeding schemes, food contact surfaces have been shown to be potential vehicles of foodborne pathogens. The aim of this study was to assess the cleanliness of the surfaces that come into contact with food that is provided to children through the National School Nutrition Programme in central South Africa. In each school under study, microbiological samples were collected from the preparation surface and the dominant hand and apron of the food handler. The samples were analysed for total viable counts, coliforms, Escherichia coli, Staphylococcus aureus and yeasts and moulds. The criteria specified in the British Columbia Guide for Environmental Health Officers were used to evaluate the results. Total viable counts were high for all surfaces, with the majority of colonies being too numerous to count (over 100 colonies per plate. Counts of organisms were relatively low, with 20% of the surfaces producing unsatisfactory enumeration of S. aureus and E. coli and 30% unsatisfactory for coliforms. Yeast and mould produced 50% and 60% unsatisfactory counts from preparation surfaces and aprons, respectively. Statistically significant differences could not be established amongst microbial counts of the surfaces, which suggests cross-contamination may have occurred. Contamination may be attributed to foodstuffs and animals in the vicinity of the preparation area rather than to the food handlers, because hands had the lowest counts of enumerated organisms amongst the analysed surfaces.

Electrolysis of polluting wastes: I - Wastewater from a seasoning freeze-drying industry

OpenAIRE

Angelis, Dejanira F. de; Corso, Carlos R.; Bidoia, Ederio D.; Moraes, Peterson B.; Domingos, Roberto N.; Rocha-Filho, Romeu C.

1998-01-01

Wastewater from a seasoning freeze-drying industry was electrolysed to increase its biodegradability. Stainless-steel electrodes were used at 9.09 A/m², for up to 80 min. Conductivity, pH, biochemical (BOD) and chemical (COD) oxygen demands, Daphnia similis acute toxicity bioassays, and bacteria counting through the plate count agar method were determined after different times of electrolysis. The results (e.g. higher BOD and lower COD) showed that the biodegradability of the wastewater was s...

Isolation and characterization of pigmented algicidal bacteria from seawater

Science.gov (United States)

Shaima, A.; Gires, U.; Asmat, A.

2014-09-01

Some dinoflagellate species are toxic and widely distributed in Malaysian marines ecosystems. They can cause many problems to aquatic life due to the production of various potential and natural toxins that accumulate in filter feeding shellfish and cause food poisoning to human. In recent decades, bacteria have been widely used as a biological control against these harmful algae. In the present study, pigmented bacteria isolated from marine water of Port Dickson beach was studied for their anti-algal activity towards toxic dinoflagellate Alexandrium minutum. Four isolates were studied and only one was capable of inhibiting algal growth when treated with bacterial culture. The algilytic effect on dinoflagellate was evaluated based on direct cell count under the microscope. Results showed that only isolate Sdpd-310 with orange colour has an inhibitory effect on A. minutum growth. This study demonstrated the rapid algicidal activity of a marine pigmented bacteria against the toxic dinoflagellate A. minutum.

Aerobic Mesophilic, Coliform, Escherichia coli, and Staphylococcus aureus Counts of Raw Meat from the Formal and Informal Meat Sectors in South Africa.

Science.gov (United States)

Jaja, Ishmael Festus; Green, Ezekiel; Muchenje, Voster

2018-04-21

Foodborne disease (FBD) is a global public health concern, and foods from animal sources have been associated with outbreaks of food-related illness. In this study, animal carcasses from the two abattoirs (HT1 and HT2) in the formal meat sector (FMS) and slaughter points in the informal meat sector (INMS) were examined at two stages of slaughter (before washing and after washing) for aerobic colony counts (ACC) and total viable count (TCC), as well as Escherichia coli and Staphylococcus aureus count. At each stage, carcasses were sampled by swabbing at the neck, brisket, flank, and rump. ACC for beef, mutton, and pork carcasses at HT1 and HT2 before washing were between 2.5⁻5.8, 2.2⁻4.7, and 2.7⁻3.7 mean log CFU/cm², respectively, and TCC count before washing was highest on the neck of cattle (6.3 ± 2.4) and after washing was highest on the perineal of sheep (5.7 ± 6.9). In the INMS, TCC count was highest on the brisket (6.9 ± 3.2) and in the neck (5.5 ± 2.4). Higher ACC values of 6.2⁻6.7 mean log CFU/cm² were obtained in the INMS. The highest count for E. coli (4.2 mean log CFU/cm²) after washing was in the neck, while the highest count for S. aureus (4.0 mean log CFU/cm²) was in the flank. All bacteria count in the INMS exceeded acceptable limits, and washing did not significantly reduce microbial load in meat in the FMS and INMS. Bacteria count in the FMS and INMS exceeded acceptable standards. However, meat processed in the INMS poses a more significant risk of FBD to consumers.

Effective Parameters on Increasing Filamentous Bacteria and Their Effects on Membrane Fouling in MBR

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Hossein Hazrati

2013-03-01

Full Text Available Over 90 percent of the wastewater treatment plants in Iran use activated sludge process. Due to increase in organic loading rates, most of these plants do not have appropriate performance. For upgrading these systems and decreasing production of the excess sludge, a UASB reactor can be used as pretreatment for decreasing the organic loading prior to the activated sludge system. Also for improving the effluent quality, a membrane can be replaced for secondary sedimentation tank, i.e. changing activated sludge to membrane bioreactor. In this study, the effect of significant changes in feed composition, due to the introduction of UASB reactor; have been investigated on the population of filamentous bacteria, COD and TS removal efficiency and membrane fouling. The results showed that the population of filamentous bacteria increased rapidly from 5 to 100 Count/µL. However, this increase does not have considerable effect on membrane fouling. With increasing MLSS concentration, the number of filamentous bacteria increased from 100 to 400Count/µL. As a result, the trans membrane pressure was raised from 1.5 to 3kpa and overall membrane resistance was increased against the effluent flux. For reducing the filamentous bacteria, a dose of 20 g Cl2 /Kg MLSS was added in few intervals for two days. It was also found the number of filamentous bacteria decreased from 400 to 100 after 5 days without decreasing the other microorganisms’ population significantly. The trans membrane pressure was also retained without any further increase.

[Phylogenetic diversity and cold-adaptive hydrolytic enzymes of culturable psychrophilic bacteria associated with sea ice from high latitude ocean, Artic].

Science.gov (United States)

Yu, Yong; Li, Hui-Rong; Chen, Bo; Zeng, Yin-Xin; He, Jian-Feng

2006-04-01

The phylogenetic diversity of culturable psychrophilic bacteria associated with sea ice from high latitude sea (77 degrees 30'N - 81 degrees 12'N), Canadian Basin and Greenland sea Arctic, was investigated. A total of 37 psychrophilic strains were isolated using three different methods of ( i ) spread plate method: 100 microL of each dilution ice-melt sample was spreaded onto the surface of Marine 2216 agar (DIFCO laboratories, Detroit, MI) and incubated for 2 to 6 weeks at 4 degrees C; ( ii ) bath culture and spread plate method: 1 mL of sample was added to 9mL of NSW (unamended natural seawater, 0.2 microm prefiltered and autoclaved) and incubated for 1 months at - 1 degrees C, then spread plate method was used to isolate bacterial strains from the pre-cultured samples; ( iii ) cold shock, bath culture and spread plate method: samples were exposed to - 20 degrees C for 24h, then bacterial strains isolated by bath culture and spread plate method under aerobic conditions. Nearly half of psychrophilic strains are isolated by using method iii . 16S rDNA nearly full-length sequence analysis reveal that psychrophilic strains fall in two phylogenetic divisions, gamma-proteobacteria (in the genera Colwellia, Marinobacter, Shewanella, Thalassomonas, Glaciecola, Marinomonas and Pseudoalteromonas) and Cytophaga-Flexibacter-Bacteroides (in the genera Flavobacterium and Psychroflexus). Nine of bacterial isolates (BSi20007, BSi20497, BSi20517, BSi20537, BSi20170, BSi20001, BSi20002, BSi20675 and BSi20101) quite likely represent novel species (16S rDNA sequence similarity below 97%). One of strains (BSi20002) from Canadian Basin shows 100% sequence similarity to the Antarctic Weddell sea ice isolate Marinobacter sp. ANT8277, suggesting bacteria may have a bipolar distribution at the species level. AF283859 sequences were submitted to the BLAST search program of the National Center for Biotechnology Information website (NCBI, http://www. ncbi. nlm.nih. gov). Twenty sequences

Comparing Methods of Separating Bacterial Biofilms on the Surface of Water Transportation Pipes and Equipment of Milking in the Farms

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setareh nabizadeh

2016-08-01

Full Text Available Introduction Bacterial biofilms can be both useful and harmful based on their combination and locations. Biofilm formation occurs as a stepwise process. Their formation in liquid transportation pipes used for milking system and drinking water in animal farms may create some problems and is a potential source of pollution. Speed of biofilm formation depends on many factors including: construction and functional characteristics of bacteria, the composition and culture conditions such as temperature and substratum. In this research the Bacillus subtillis bacteria with special characteristics was selected due to its capability for biofilm creation. Bacillus subtillis bacteria is mobility and a stronger connection than other bacteria levels are created. In the research conducted in the biofilm there are many resources on biofilm formation by Bacillus subtillis bacteria. Bacillus subtillis is saprophytic in the soil, water and air. There is also the ability to form spores of Bacillus subtillis. Materials and Methods Firstly the possibility of creating biofilms on different Plastic (polyvinilchlorid, polypropylene, polyethylengelycole, alluminum and glass surfaces in three temperatures of 4°C, 30°C and 37°C were studied. Two different methods of biofilms separation including separating swap and vortex were tested and their efficienceies were calculated. After biofilm formation on parts of the vortex separation method after washing parts in sterile conditions in a tube containing normal saline for 4 minutes was vortex. The bacterial suspension decreasing dilution series was created. Pour plate in medium using agar plate count agar and was cultured at 30°C for 24-48 hours. Numbers of colonies were counted. The numbers of biofilm cells were calculated. In swap method after biofilm formation on parts using a cotton swap was isolated biofilms. The swap was transferred to tube containing normal saline and the bacterial suspension decreasing dilution

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

Science.gov (United States)

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.

Compton suppression gamma-counting: The effect of count rate

Science.gov (United States)

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.

EcoCount

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Phillip P. Allen

2014-05-01

Full Text Available Techniques that analyze biological remains from sediment sequences for environmental reconstructions are well established and widely used. Yet, identifying, counting, and recording biological evidence such as pollen grains remain a highly skilled, demanding, and time-consuming task. Standard procedure requires the classification and recording of between 300 and 500 pollen grains from each representative sample. Recording the data from a pollen count requires significant effort and focused resources from the palynologist. However, when an adaptation to the recording procedure is utilized, efficiency and time economy improve. We describe EcoCount, which represents a development in environmental data recording procedure. EcoCount is a voice activated fully customizable digital count sheet that allows the investigator to continuously interact with a field of view during the data recording. Continuous viewing allows the palynologist the opportunity to remain engaged with the essential task, identification, for longer, making pollen counting more efficient and economical. EcoCount is a versatile software package that can be used to record a variety of environmental evidence and can be installed onto different computer platforms, making the adoption by users and laboratories simple and inexpensive. The user-friendly format of EcoCount allows any novice to be competent and functional in a very short time.

RBC count

Science.gov (United States)

... by kidney disease) RBC destruction ( hemolysis ) due to transfusion, blood vessel injury, or other cause Leukemia Malnutrition Bone ... slight risk any time the skin is broken) Alternative Names Erythrocyte count; Red blood cell count; Anemia - RBC count Images Blood test ...

Contaminant bacteria in traditional-packed honey

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Hening Tjaturina Pramesti

2007-03-01

Full Text Available Honey may be contaminated by microorganisms during its harvesting, processing, and packaging. Honey selected for clinical purposes must safe, sterile, and contain antimicrobial activity, so it must be evaluated using laboratory testing. The aim of this descriptive laboratory study was to isolate and identify the bacterial contaminant in the traditional-packed honey dealing with the use of honey for medical purposes. the colony forming units of honey sample cultured on blood agar were counted using Stuart bacterial colony counter. The suspected bacterial colonies were isolated and identified based on cultural morphology characteristics. The isolates of suspected bacterial colonies were stained according to Gram and Klein method and then were examined by the biochemical reaction. The results showed that there were two contaminant bacteria. Gram-positive cocci which were presumptively identified as coagulase-negative Staphylococci and gram-positive rods which were presumptively identified as Bacillus subtilis. In conclusion, the contaminant bacteria were regarded as low pathogen bacteria. The subtilin enzyme of B subtilis may cause an allergic reaction and coagulase-negative Staphylococci, Staphylococcus epidermidis is also an opportunist pathogen. Inevitably, for medical purposes, traditional-packed honey must be well filtered, water content above 18%, and standardized sterilization without loss of an antibacterial activity or change in properties.

Impact of fertilizing with raw or anaerobically digested sewage sludge on the abundance of antibiotic-resistant coliforms, antibiotic resistance genes, and pathogenic bacteria in soil and on vegetables at harvest.

Science.gov (United States)

Rahube, Teddie O; Marti, Romain; Scott, Andrew; Tien, Yuan-Ching; Murray, Roger; Sabourin, Lyne; Zhang, Yun; Duenk, Peter; Lapen, David R; Topp, Edward

2014-11-01

The consumption of crops fertilized with human waste represents a potential route of exposure to antibiotic-resistant fecal bacteria. The present study evaluated the abundance of bacteria and antibiotic resistance genes by using both culture-dependent and molecular methods. Various vegetables (lettuce, carrots, radish, and tomatoes) were sown into field plots fertilized inorganically or with class B biosolids or untreated municipal sewage sludge and harvested when of marketable quality. Analysis of viable pathogenic bacteria or antibiotic-resistant coliform bacteria by plate counts did not reveal significant treatment effects of fertilization with class B biosolids or untreated sewage sludge on the vegetables. Numerous targeted genes associated with antibiotic resistance and mobile genetic elements were detected by PCR in soil and on vegetables at harvest from plots that received no organic amendment. However, in the season of application, vegetables harvested from plots treated with either material carried gene targets not detected in the absence of amendment. Several gene targets evaluated by using quantitative PCR (qPCR) were considerably more abundant on vegetables harvested from sewage sludge-treated plots than on vegetables from control plots in the season of application, whereas vegetables harvested the following year revealed no treatment effect. Overall, the results of the present study suggest that producing vegetable crops in ground fertilized with human waste without appropriate delay or pretreatment will result in an additional burden of antibiotic resistance genes on harvested crops. Managing human exposure to antibiotic resistance genes carried in human waste must be undertaken through judicious agricultural practice. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

The Effect of Some Physical and Chemical Parameters on Regrowth of Aeromonas Bacterium and Heterotrophic Bacteria in Isfahan Drinking Water System

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Sina Dobaradaran

2006-03-01

Full Text Available Aeromonas is one the gram – negative , non spor – formating rod shaping , facultatively anaerobic and opportunistic bactria that can cause systematic infections, leision and diarrhoea in human. Fairly high bactrial population in distribution system is not only of concern because of affecting consumer health but also it makes it difficult to enumerate coliform bacterium indicator. So, the relationships between aeromonas and heterotrophic bacteria growth with pH, temperature, turbidity, free residual cholornie and DO were determined in this study. ADA- V media was used in presumptive stage to count aeromonas bacteria for the first time in Iran on the basis of 1605 EPA (2001 method and used oxidase tests, trehalose fermentation and indol test in confirmative stage. R2A media was used to count HPC bacteria and other factors measured on the basis of standards. The results showed that positive cases of aeromonas bacteria and HPC increase in higher temperature and turbidity and lower pH. In contrast, positive cases of aeromonas bacteria and HPC decrease while free residual cholorine and DO increase. In addition,no positive case of aeromonas was observed in more than 0.2 mg/L concentration of free residual cholorine.

Effect of face washing with soap and water and cleaning with antiseptics on upper-lid bacteria of surgical eye patients.

Science.gov (United States)

Bekibele, Charles O; Kehinde, Aderemi O; Ajayi, Benedictus G K

2010-12-01

To determine the effect of face washing with soap and water and cleaning with povidone iodine and cetrimide/chlorhexidine gluconate (Savlon) on upper-lid bacteria. Prospective, nonrandomized clinical trial. Eighty patients attending the Eye Clinic, University College Hospital, Ibadan, Nigeria. Eighty patients assigned to 4 groups had swabs of the upper eyelid skin taken before and after face wash with soap and water, and cleansing with Savlon and 5% povidone iodine. Specimens were cultured and Gram stained. Bacterial counts were carried out using standard techniques. Face washing with soap and water increased the proportion of patients with bacterial isolates from 80.0% to 87.5%. The average colony count increased from 187.1 to 318.5 colony units per mL (p = 0.02). Application of 5% povidone iodine without face washing with soap and water reduced the proportion of patients with bacterial isolates from 82.6% (mean count 196.5) to 28.6% (mean count 34.1)(p = 0.001); in comparison, the application of 5% povidone iodine after face washing with soap and water reduced the proportion from 71.4% (mean count 133.9) to 40.0% (mean count 69.0)(p = 0.01). Application of Savlon without face washing with soap and water reduced the proportion of patients with bacterial isolates from 100% (mean count 310.9) to 41.2% (mean count 19.8)(p = 0.004) compared with the application after face washing, which reduced the proportion from 89.5% (mean count 240.3) to 41.2% (mean count 82.9)(p = 0.02). Both povidone and Savlon are effective in reducing periocular bacteria in an African setting. Prior face washing with soap and water had no added benefit in reducing bacterial colony count.

Contamination of water wells in Khoms city with pathogenic coliform bacteria

International Nuclear Information System (INIS)

Mahjoub, Tariq M.; Buazzi, Mahmoud M.; Jamil, Ahmad Y.

2007-01-01

240 Samples from 60 water wells in and around the area of city of Khoms city (in northwestern Libya) were analyzed over four successive seasons for count of faecal Coliform bacteria, of which antibiotic resistance was later assessed. Standard methods were used for analysis of faecla coliform bacteria. Water wells contained varying levels of faecal coliform bacteria ranging from a Most Probable Number of 0 to 1.8 x 10 3 cfu/100 ml, with zero isolates at autumn and winter seasons, while wells dug at private farms had the highest percentage of contamination, reaching 56.6% of wells in autumn, and also had the highest number of faecal coliform isolates, 1,8x10 3 cfu/100 ml, in spring and summer seasons, strains of isolated Escherichia coli were most sensitive to chloramphenicol, and most resistant to tetracycline. (author)

EVALUATION OF THE TEA TREE OIL ACTIVITY TO ANAEROBIC BACTERIA--IN VITRO STUDY.

Science.gov (United States)

Ziółkowska-Klinkosz, Marta; Kedzia, Anna; Meissner, Hhenry O; Kedzia, Andrzej W

2016-01-01

The study of the sensitivity to tea tree oil (Australian Company TTD International Pty. Ltd. Sydney) was carried out on 193 strains of anaerobic bacteria isolated from patients with various infections within the oral cavity and respiratory tracts. The susceptibility (MIC) of anaerobes was determined by means of plate dilution technique in Brucella agar supplemented with 5% defibrinated sheep blood, menadione and hemin. Inoculum contained 10(5) CFU per spot was cultured with Steers replicator upon the surface of agar with various tea tree oil concentrations or without oil (anaerobes growth control). Incubation the plates was performed in anaerobic jars under anaerobic conditions at 37 degrees C for 48 h. MIC was defined as the lowest concentrations of the essential oil completely inhibiting growth of anaerobic bacteria. Test results indicate, that among Gram-negative bacteria the most sensitive to essential oil were strains of Veillonella and Porphyromonas species. Essential oil in low concentrations (MIC in the range of = 0.12 - 0.5 mg/mL) inhibited growth of accordingly 80% and 68% strains. The least sensitive were strains of the genus Tannerella, Parabacteroides and Dialister (MIC 1.0 - 2.0 mg/mL). In the case of Gram-positive anaerobic bacteria the tea tree oil was the most active to strains of cocci of the genus Anaerococcus and Ruminococcus (MIC in range = 0.12 - 0.5 mg/mL) or strains of rods of the genus Eubacterium and Eggerthella (MIC = 0.25 mg/mL). Among Gram-positive rods the least sensitive were the strains of the genus Bifidobacterium ( MIC = 2.0 mg/mL). The tea tree oil was more active to Gram-positive than to Gram-negative anaerobic bacteria.

Utilization of Vinegar for Isolation of Cellulose Producing Acetic Acid Bacteria

International Nuclear Information System (INIS)

Aydin, Y. Andelib; Aksoy, Nuran Deveci

2010-01-01

Wastes of traditionally fermented Turkish vinegar were used in the isolation of cellulose producing acetic acid bacteria. Waste material was pre-enriched in Hestrin-Schramm medium and microorganisms were isolated by plating dilution series on HS agar plates The isolated strains were subjected to elaborate biochemical and physiological tests for identification. Test results were compared to those of reference strains Gluconacetobacter xylinus DSM 46604, Gluconacetobacter hansenii DSM 5602 and Gluconacetobacter liquefaciens DSM 5603. Seventeen strains, out of which only three were found to secrete the exopolysaccharide cellulose. The highest cellulose yield was recorded as 0.263±0.02 g cellulose L -1 for the strain AS14 which resembled Gluconacetobacter hansenii in terms of biochemical tests.

Incorporation of nisin in natural casing for the control of spoilage microorganisms in vacuum packaged sausage

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Joyce Regina de Barros

2010-12-01

Full Text Available This study aimed to evaluate the effectiveness of natural casing treatment with nisin and phosphoric acid on control of spoilage microorganisms in vacuum packaged sausages. Ovine casings were dipped in the following baths: 1 0.1% food grade phosphoric acid; 2 5.0 mg/L nisin; 3 0.1% phosphoric acid and 5.0 mg/L nisin; and 4 sterile water (control. The sausages were produced in a pilot plant, stuffed into the pretreated natural casings, vacuum packaged and stored at 4 and 10 °C for 56 days. The experiments were performed according to a full factorial design 2³, totalizing 8 treatments that were repeated in 3 blocks. Aerobic plate counts and lactic acid bacteria analysis were conducted at 1, 14, 28, 42 and 56 days of storage. Treatment of casings with phosphoric acid 0.1% alone did not inhibit the growth of lactic acid bacteria and reduced the aerobic plate count by 1 log. The activity of nisin against lactic acid bacteria was enhanced by the addition of phosphoric acid, demonstrating a synergistic effect. Furthermore nisin activity was more evident at lower storage temperature (4 ºC. Therefore treatment of the natural casings with nisin and phosphoric acid, combined with low storage temperature, are obstacles that present a potential for controlling the growth of lactic acid bacteria in vacuum packaged sausage.

Preliminary Identification of the Microbial Fuel Cell Bacteria Communities in Sewage

International Nuclear Information System (INIS)

Zain, S.M; Hashim, R.; Roslani, N.S.; Suja, F.; Basri, N.E.A.; Anuar, N.; Daud, W.R.W.

2011-01-01

This study aimed to determine the types of bacteria exist in wastewater that contribute to generate electricity and simultaneously remove carbon and nitrogen. The method used was Fluorescence In Situ Hybridization (FISH) to detect the bacteria group while Polymerase Chain Reaction (PCR) was used to confirm the observation made using FISH. A biochemical identification using BIOLOG GEN III MICROPLATE TM also was used . The samples were cultured on nutrient agar plate to identify the morphology of the bacteria. The result showed that 21 isolates from three different locations at the activated sludge treatment plant with six, eleven and four strains for raw sewage, aeration tank and returned activated sludge samples, respectively. Preliminary identification does not give a good match but only showed the existence of ammonia-oxidizing bacteria (FISH) and Kurtia Gibsoni (BIOLOG) from aeration tank : Bacillus sp (PCR) and Bacillus Pseudomycoides (BIOLOG) from returned activated sludge. The maximum power density generated using returned activated sludge was 9.053 mW/ cm 2 , with 26.8 % COD removal and 40 % TKN removal (author)

Antimicrobial peptides secreted by equine mesenchymal stromal cells inhibit the growth of bacteria commonly found in skin wounds.

Science.gov (United States)

Harman, Rebecca M; Yang, Steven; He, Megan K; Van de Walle, Gerlinde R

2017-07-04

The prevalence of chronic skin wounds in humans is high, and treatment is often complicated by the presence of pathogenic bacteria. Therefore, safe and innovative treatments to reduce the bacterial load in cutaneous wounds are needed. Mesenchymal stromal cells (MSC) are known to provide paracrine signals that act on resident skin cells to promote wound healing, but their potential antibacterial activities are not well described. The present study was designed to examine the antibacterial properties of MSC from horses, as this animal model offers a readily translatable model for MSC therapies in humans. Specifically, we aimed to (i) evaluate the in vitro effects of equine MSC on the growth of representative gram-negative and gram-positive bacterial species commonly found in skin wounds and (ii) define the mechanisms by which MSC inhibit bacterial growth. MSC were isolated from the peripheral blood of healthy horses. Gram-negative E. coli and gram-positive S. aureus were cultured in the presence of MSC and MSC conditioned medium (CM), containing all factors secreted by MSC. Bacterial growth was measured by plating bacteria and counting viable colonies or by reading the absorbance of bacterial cultures. Bacterial membrane damage was detected by incorporation of N-phenyl-1-naphthylamine (NPN). Antimicrobial peptide (AMP) gene and protein expression by equine MSC were determined by RT-PCR and Western blot analysis, respectively. Blocking of AMP activity of MSC CM was achieved using AMP-specific antibodies. We found that equine MSC and MSC CM inhibit the growth of E. coli and S. aureus, and that MSC CM depolarizes the cell membranes of these bacteria. In addition, we found that equine MSC CM contains AMPs, and blocking these AMPs with antibodies reduces the effects of MSC CM on bacteria. Our results demonstrate that equine MSC inhibit bacterial growth and secrete factors that compromise the membrane integrity of bacteria commonly found in skin wounds. We also identified

Rapid detection of microbial contamination in grape juice by flow cytometry

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Marielle Bouix

1999-03-01

Full Text Available This study presents an application of flow cytometry to evaluate rapidly the viable micro-organisms in grape juice. In this method, viable cells are firstly specitically labelled with a fluorescent reagent. The sample is then injected into the flow cytometer where the labelled micro-organisms are individually illuminated by a laser beam. The emission of fluorescence is measured. The system counts the number of fluorescent events and prints out a histogram of the fluorescence intensity which is characteristic of the micro-organism being analysed. In laboratory conditions, preliminary trials have been undertaken with an artificially inoculated grape juice with pure yeast and bacteria cultures. This method succeeded in counting simultaneously yeasts and bacteria within 15 minutes, with a high degree of sensitivity, 5.103 yeasts perml and 5.104 bacteria per ml. This technique can also be applied to the detection of mould contamination and the test has been done with Botrytis spores. The method makes direct cell counts possible and is capable of analysing 30 samples per hour. It can be automatised and easily used in industrial laboratory. During the last harvest, more than a thousand of must samples were controled using this technique. The results let to determine the yeast contamination level of a grape juice tank even before unloading. The results obtained by flow cytometry were compared to the plate count reference method. The correlation between cytometry and count by plate culture was 99 p. cent for the threshold of 5.1 04 yeasts/ml which seemed to point out a high contamination. By using this flow cytometry method during the harvest period, the results were supplied in real time. This allowed a rapid selection of the musts, depending upon the scale of their contamination and improved the quality of the wine by corrective actions.

MICROBIAL QUALITY OF HONEY MIXTURE WITH POLLEN

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Ján Mareček

2011-02-01

Full Text Available Normal 0 21 MicrosoftInternetExplorer4 The aim of this study was evaluation of microbial quality in raw materials (honey, pollen and evaluation of microbial quality in honey mixture with pollen (2.91 % and 3.85 % and also dynamics of microbial groups in honey mixtures with pollen after 14 days storage at the room temperature (approximately 25 °C and in cold store (8 °C. We used dilution plating method for testing of samples. Detections of total plate microbial count (aerobic and anaerobic microorganisms, sporulating bacteria, coliform bacteria, Bifidobacterium sp., Lactobacillus sp. and microscopic fungi were performed. In general, counts of microorganisms decreased in honey mixture with pollen compared to raw pollen and these counts increased compared to natural honey. Total plate count was 5.37 log KTJ.g-1 in pollen; 1.36 log KTJ.g-1 in honey; 2.97 log KTJ.g-1 in honey mixture with 2.91 % pollen and 2.04 log KTJ.g-1 in honey mixture with 3.85 % pollen. Coliform bacteria were detected in pollen (1.77 log KTJ.g-1. Then, we found coliform bacteria in one sample of honey mixtures with pollen (2.91 % - 1.00 log KTJ.g-1.Bifidobacterium species were detected only in raw pollen. We did not findLactobacillus sp. in any of the samples. Microscopic fungi were detected on two cultivating media. Yeasts were present in pollen sample (average 5.39 log KTJ.g-1, honey mixture with 2.91 % pollen (average 2.51 log KTJ.g-1 and honey mixture with 3.85 % pollen (average 1.58 log KTJ.g-1. Filamentous microscopic fungi were detectable in pollen (average 3.38 log KTJ.g-1, in honey (only on one medium: 1.00 log KTJ.g-1, in honey mixture with 2.91 % pollen (average 1.15 log KTJ.g-1 and in honey mixture with 3.85 % pollen (1.71 %. Raw pollen contained microscopic fungi as Absidiasp., Mucor sp., Alternaria sp. andEmericella nidulans. Honey mixture with 2.91 % pollen after storage (14 days contained lower microbial counts when compared with the sample

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

International Nuclear Information System (INIS)

Scotter, S.L.; Wood, R.; McWeeny, D.J.

1990-01-01

A study to evaluate the potential of the Limulus amoebocyte lysate (LAL) test in conjunction with a Gram negative bacterial (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 0 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 amd during subsequent chilled storage also aided differentiation of irradiated and unirradiated birds. Large numbers of yeast and Gram positive cocci were isolated from irradiated carcasses whereas Gram negative oxidative rods were the predominant spoilage flora on unirradiated birds. (author)

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

Energy Technology Data Exchange (ETDEWEB)

Scotter, S L; Wood, R; McWeeny, D J [Ministry of Agriculture, Fisheries and Food, Norwich (UK). Food Science Lab.

1990-01-01

A study to evaluate the potential of the Limulus amoebocyte lysate (LAL) test in conjunction with a Gram negative bacterial (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{sup 0}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 amd during subsequent chilled storage also aided differentiation of irradiated and unirradiated birds. Large numbers of yeast and Gram positive cocci were isolated from irradiated carcasses whereas Gram negative oxidative rods were the predominant spoilage flora on unirradiated birds. (author).

Ammonia production by human faecal bacteria, and the enumeration, isolation and characterization of bacteria capable of growth on peptides and amino acids

Directory of Open Access Journals (Sweden)

Richardson Anthony J

2013-01-01

Full Text Available Abstract Background The products of protein breakdown in the human colon are considered to be detrimental to gut health. Amino acid catabolism leads to the formation of sulfides, phenolic compounds and amines, which are inflammatory and/or precursors to the formation of carcinogens, including N-nitroso compounds. The aim of this study was to investigate the kinetics of protein breakdown and the bacterial species involved. Results Casein, pancreatic casein hydrolysate (mainly short-chain peptides or amino acids were incubated in vitro with suspensions of faecal bacteria from 3 omnivorous and 3 vegetarian human donors. Results from the two donor groups were similar. Ammonia production was highest from peptides, followed by casein and amino acids, which were similar. The amino acids metabolized most extensively were Asp, Ser, Lys and Glu. Monensin inhibited the rate of ammonia production from amino acids by 60% (P = 0.001, indicating the involvement of Gram-positive bacteria. Enrichment cultures were carried out to investigate if, by analogy with the rumen, there was a significant population of asaccharolytic, obligately amino acid-fermenting bacteria (‘hyper-ammonia-producing’ bacteria; HAP in the colon. Numbers of bacteria capable of growth on peptides or amino acids alone averaged 3.5% of the total viable count, somewhat higher than the rumen. None of these were HAP, however. The species enriched included Clostridium spp., one of which was C. perfringens, Enterococcus, Shigella and Escherichia coli. Conclusions Protein fermentation by human faecal bacteria in the absence of sugars not only leads to the formation of hazardous metabolic products, but also to the possible proliferation of harmful bacteria. The kinetics of protein metabolism were similar to the rumen, but HAP bacteria were not found.

A position sensitive parallel plate avalanche fission detector for use in particle induced fission coincidence measurements

NARCIS (Netherlands)

Plicht, J. van der

1980-01-01

A parallel plate avalanche detector developed for the detection of fission fragments in particle induced fission reactions is described. The active area is 6 × 10 cm2; it is position sensitive in one dimension with a resolution of 2.5 mm. The detector can withstand a count rate of 25000 fission

Correlation between slaughter practices and the distribution of Salmonella and hygiene indicator bacteria on pig carcasses during slaughter.

Science.gov (United States)

Biasino, W; De Zutter, L; Mattheus, W; Bertrand, S; Uyttendaele, M; Van Damme, I

2018-04-01

This study investigated the distribution of hygiene indicator bacteria and Salmonella on pig carcasses. Moreover, the relation between hygiene indicator counts and Salmonella presence as well as associations between specific slaughter practices and carcass contamination were determined for each carcass area. Seven Belgian pig slaughterhouses were visited three times to swab five randomly selected carcasses at nine different areas, after evisceration and trimming. Information about slaughter practices was collected using a questionaire. In all samples, the E. coli and Salmonella presence was analyzed and Enterobacteriaceae and total aerobic bacteria were quantified. Average total aerobic counts ranged from 3.1 (loin, pelvic duct, ham) to 4.4 log 10  CFU/cm 2 (foreleg). Median Enterobacteriaceae numbers varied between 0.4 (ham) an 1.8 log 10  CFU/cm 2 (foreleg). E. coli and Salmonella presence ranged from 15% (elbow) to 89% (foreleg) and 5% (elbow) to 38% (foreleg), respectively. Positive relations were found between hygiene indicator counts and Salmonella presence at the head, sternum, loin and throat. Several slaughter practices, such as splitting the head and incising tonsils, were associated with higher levels of hygiene indicator bacteria and Salmonella. These findings can be used to educate slaughterhouse personnel and estimate the public health risk involved in consumption of different pork cuts. Copyright © 2017 Elsevier Ltd. All rights reserved.

Counting statistics in low level radioactivity measurements fluctuating counting efficiency

International Nuclear Information System (INIS)

Pazdur, M.F.

1976-01-01

A divergence between the probability distribution of the number of nuclear disintegrations and the number of observed counts, caused by counting efficiency fluctuation, is discussed. The negative binominal distribution is proposed to describe the probability distribution of the number of counts, instead of Poisson distribution, which is assumed to hold for the number of nuclear disintegrations only. From actual measurements the r.m.s. amplitude of counting efficiency fluctuation is estimated. Some consequences of counting efficiency fluctuation are investigated and the corresponding formulae are derived: (1) for detection limit as a function of the number of partial measurements and the relative amplitude of counting efficiency fluctuation, and (2) for optimum allocation of the number of partial measurements between sample and background. (author)

Characteristic of cow milk dadih using starter of probiotic of lactic acid bacteria

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Sri Usmiati

2011-06-01

Full Text Available Dadih is an original dairy product from West Sumatera processed traditionally. It is a spontaneous fermentation of buffalo milk at room temperature for 48 hours in a bamboo tube, has no standard of processing and quality. Dadih is potentially to be develop into probiotic products (functional food that can be enjoyed by the public widely. Development of cow's milk dadih is necessary since buffalo milk is available only in certain area. Product and characteristic information of cow milk dadih using probiotic of lactic acid bacteria starter has not been known. The research objective was to determine the characteristics of cow milk dadih that used starter of probiotic lactic acid bacteria during storage at room temperature (27oC and cold temperature (4oC. The study was designed using a factorial randomized block design pattern 12x3 at room temperature and 12X4 at cold temperatures, with the number of repeatation of 3 times. Treatment consisted of: (i starter formula (A using a single bacterium or a combination of Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium longum, and (ii storage time (B. Observed parameters included pH value, titrable acidity, the total plate count, and in-vitro probiotic testing (bacterial resistance to bile salts and low pH of cow milk dadih. The results showed that L. acidophilus early exponential phase was at the hour 3rd, L. casei at the hour 4th and B. longum on the 3rd of which is used as the optimum time of mixing two or more bacteria in the manufacture of cow milk dadih. The volume of starter used was 3% with time fermentation of 48 hours at room temperature (27-30oC. Cow milk dadih that was stored for 7 days at room temperature (27-30oC and for 21 days at cold temperatures (4-10oC was able to maintain viability of bacteria to bile salts and low pH at 1010-1012 cfu/ml with percentage resistance varied. The cow milk dadih using a combination starter of B. longum with other probiotics on the

Cold plate

Energy Technology Data Exchange (ETDEWEB)

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.

Biofilm Formation by Bacteria Isolated from Intravenous Catheters

Directory of Open Access Journals (Sweden)

Sina Hedayati

2015-10-01

Full Text Available Background: Reports on the association of nosocomial bacterial infections with indwelling medical devices such as intravenous catheters (IVC has increased in recent years. The potential to form biofilm on these devices seems to be the main reason for establishment of such infections. The aim of this study was to measure the potential of biofilm formation by bacterialisolates from IVCs.Methods: Seventy-one IVCs were collected from hospitalized patients in ICU, NICU, hematology and oncology wards at Taleghani Hospital from Jan 2010 to Jan 2011. The bacterial isolates were identified using the standard biochemical tests and the potential to form biofilms was determined by the microtiter plate assay method (MTP and colony morphology using Congo red agar plates (CRA.Results: Overall, 54 (71% IVCs were colonized and 76 bacteria were isolated among which, 64 (84.2% were coagulase negative staphylococci (CoNS, 3 (3.9% S. aureus, 3 (3.9% Enterococcus spp., 2 (2.6% E. coli and 4 (5.3% were miscellaneous isolates not further identified. Among the CoNS, biofilm formation was observed in 68.7% and 82.8% of bacteriausing MTP and CRA methods, respectively. S. aureus and E. coli isolates also were biofilm producers but Enterococcus and other unknown isolates were biofilm negative.Conclusions: Our results confirm that the prevalent biofilm forming bacteria on IVCs were CoNS and that was the reason for high rates of nosocomial infections.

A direct pre-screen for marine bacteria producing compounds inhibiting quorum sensing reveals diverse planktonic bacteria that are bioactive.

Science.gov (United States)

Linthorne, Jamie S; Chang, Barbara J; Flematti, Gavin R; Ghisalberti, Emilio L; Sutton, David C

2015-02-01

A promising new strategy in antibacterial research is inhibition of the bacterial communication system termed quorum sensing. In this study, a novel and rapid pre-screening method was developed to detect the production of chemical inhibitors of this system (quorum-quenching compounds) by bacteria isolated from marine and estuarine waters. This method involves direct screening of mixed populations on an agar plate, facilitating specific isolation of bioactive colonies. The assay showed that between 4 and 46 % of culturable bacteria from various samples were bioactive, and of the 95 selectively isolated bacteria, 93.7 % inhibited Vibrio harveyi bioluminescence without inhibiting growth, indicating potential production of quorum-quenching compounds. Of the active isolates, 21 % showed further activity against quorum-sensing-regulated pigment production by Serratia marcescens. The majority of bioactive isolates were identified by 16S ribosomal DNA (rDNA) amplification and sequencing as belonging to the genera Vibrio and Pseudoalteromonas. Extracts of two strongly bioactive Pseudoalteromonas isolates (K1 and B2) were quantitatively assessed for inhibition of growth and quorum-sensing-regulated processes in V. harveyi, S. marcescens and Chromobacterium violaceum. Extracts of the isolates reduced V. harveyi bioluminescence by as much as 98 % and C. violaceum pigment production by 36 % at concentrations which had no adverse effect on growth. The activity found in the extracts indicated that the isolates may produce quorum-quenching compounds. This study further supports the suggestion that quorum quenching may be a common attribute among culturable planktonic marine and estuarine bacteria.

Size-dependent antibacterial activities of silver nanoparticles against oral anaerobic pathogenic bacteria.

Science.gov (United States)

Lu, Zhong; Rong, Kaifeng; Li, Ju; Yang, Hao; Chen, Rong

2013-06-01

Dental caries and periodontal disease are widespread diseases for which microorganism infections have been identified as the main etiology. Silver nanoparticles (Ag Nps) were considered as potential control oral bacteria infection agent due to its excellent antimicrobial activity and non acute toxic effects on human cells. In this work, stable Ag Nps with different sizes (~5, 15 and 55 nm mean values) were synthesized by using a simple reduction method or hydrothermal method. The Nps were characterized by powder X-ray diffraction, transmission electron microscopy and UV-vis absorption spectroscopy. The antibacterial activities were evaluated by colony counting assay and growth inhibition curve method, and corresponding minimum inhibitory concentration (MIC) against five anaerobic oral pathogenic bacteria and aerobic bacteria E. coli were determined. The results showed that Ag Nps had apparent antibacterial effects against the anaerobic oral pathogenic bacteria and aerobic bacteria. The MIC values of 5-nm Ag against anaerobic oral pathogenic bacteria A. actinomycetemcomitans, F. nuceatum, S. mitis, S. mutans and S. sanguis were 25, 25, 25, 50 and 50 μg/mL, respectively. The aerobic bacteria were more susceptible to Ag NPs than the anaerobic oral pathogenic bacteria. In the mean time, Ag NPs displayed an obvious size-dependent antibacterial activity against the anaerobic bacteria. The 5-nm Ag presents the highest antibacterial activity. The results of this work indicated a potential application of Ag Nps in the inhibition of oral microorganism infections.

Antibody Microarray for E. coli O157:H7 and Shiga Toxin in Microtiter Plates

Directory of Open Access Journals (Sweden)

Andrew G. Gehring

2015-12-01

Full Text Available Antibody microarray is a powerful analytical technique because of its inherent ability to simultaneously discriminate and measure numerous analytes, therefore making the technique conducive to both the multiplexed detection and identification of bacterial analytes (i.e., whole cells, as well as associated metabolites and/or toxins. We developed a sandwich fluorescent immunoassay combined with a high-throughput, multiwell plate microarray detection format. Inexpensive polystyrene plates were employed containing passively adsorbed, array-printed capture antibodies. During sample reaction, centrifugation was the only strategy found to significantly improve capture, and hence detection, of bacteria (pathogenic Escherichia coli O157:H7 to planar capture surfaces containing printed antibodies. Whereas several other sample incubation techniques (e.g., static vs. agitation had minimal effect. Immobilized bacteria were labeled with a red-orange-fluorescent dye (Alexa Fluor 555 conjugated antibody to allow for quantitative detection of the captured bacteria with a laser scanner. Shiga toxin 1 (Stx1 could be simultaneously detected along with the cells, but none of the agitation techniques employed during incubation improved detection of the relatively small biomolecule. Under optimal conditions, the assay had demonstrated limits of detection of ~5.8 × 105 cells/mL and 110 ng/mL for E. coli O157:H7 and Stx1, respectively, in a ~75 min total assay time.

Antibody Microarray for E. coli O157:H7 and Shiga Toxin in Microtiter Plates.

Science.gov (United States)

Gehring, Andrew G; Brewster, Jeffrey D; He, Yiping; Irwin, Peter L; Paoli, George C; Simons, Tawana; Tu, Shu-I; Uknalis, Joseph

2015-12-04

Antibody microarray is a powerful analytical technique because of its inherent ability to simultaneously discriminate and measure numerous analytes, therefore making the technique conducive to both the multiplexed detection and identification of bacterial analytes (i.e., whole cells, as well as associated metabolites and/or toxins). We developed a sandwich fluorescent immunoassay combined with a high-throughput, multiwell plate microarray detection format. Inexpensive polystyrene plates were employed containing passively adsorbed, array-printed capture antibodies. During sample reaction, centrifugation was the only strategy found to significantly improve capture, and hence detection, of bacteria (pathogenic Escherichia coli O157:H7) to planar capture surfaces containing printed antibodies. Whereas several other sample incubation techniques (e.g., static vs. agitation) had minimal effect. Immobilized bacteria were labeled with a red-orange-fluorescent dye (Alexa Fluor 555) conjugated antibody to allow for quantitative detection of the captured bacteria with a laser scanner. Shiga toxin 1 (Stx1) could be simultaneously detected along with the cells, but none of the agitation techniques employed during incubation improved detection of the relatively small biomolecule. Under optimal conditions, the assay had demonstrated limits of detection of ~5.8 × 10⁵ cells/mL and 110 ng/mL for E. coli O157:H7 and Stx1, respectively, in a ~75 min total assay time.

Bacteria in non-woven textile filters for domestic wastewater treatment.

Science.gov (United States)

Spychała, Marcin; Starzyk, Justyna

2015-01-01

The objective of this study was preliminary identification of heterotrophic and ammonia oxidizing bacteria (AOB) cell concentration in the cross-sectional profile of geotextile filters for wastewater treatment. Filters of thicknesses 3.6 and 7.2 mm, made of non-woven textile TS20, were supplied with septic tank effluent and intermittently dosed and filtered under hydrostatic pressure. The cumulative loads of chemical oxygen demand (COD) and total solids were about 1.36 and 1.06 kg/cm2, respectively. The filters under analysis reached a relatively high removal efficiency for organic pollution 70-90% for biochemical oxygen demand (BOD5) and 60-85% for COD. The ammonia nitrogen removal efficiency level proved to be unstable (15-55%). Biomass samples for dry mass identification were taken from two regions: continuously flooded with wastewater and intermittently flooded with wastewater. The culturable heterotrophic bacteria were determined as colony-forming units (CFUs) on microbiological-selective media by means of the plate method. AOB and nitrite oxidizing bacteria (NOB) were examined using the FISH technique. A relatively wide range of heterotrophic bacteria was observed from 7.4×10(5)/cm2 to 3.8×10(6)/cm2 in geotextile layers. The highest concentration of heterotrophic bacteria (3.8×10(6)/cm2) was observed in the first layer of the textile filter. AOB were identified occasionally--about 8-15% of all bacteria colonizing the last filter layer, but occasionally much higher concentrations and ammonia nitrogen efficiency were achieved. Bacteria oxidizing nitrite to nitrate were not observed. The relation of total and organic fraction of biomass to culturable heterotrophic bacteria was also found.

Characterization of specimens obtained by different sampling methods for evaluation of periodontal bacteria.

Science.gov (United States)

Okada, Ayako; Sogabe, Kaoru; Takeuchi, Hiroaki; Okamoto, Masaaki; Nomura, Yoshiaki; Hanada, Nobuhiro

2017-12-27

Quantitative analysis of periodontal bacteria is considered useful for clinical diagnosis, evaluation and assessment of the risk of periodontal disease. The purpose of this study was to compare the effectiveness of sampling of saliva, supragingival and subgingival plaque for evaluation of periodontal bacteria. From each of 12 subjects, i) subgingival plaque was collected from the deepest pocket using a sterile paper point, ii) stimulated whole saliva was collected after chewing gum, and iii) supragingival plaque was collected using a tooth brush. These samples were sent to the medical examination laboratory for quantitative analysis of the counts of three periodontal bacterial species: Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia. The proportions of these bacteria in subgingival plaque were higher than those in saliva or supragingival plaque, but lower in subgingival plaque than in saliva or supragingival plaque. In several cases, periodontal bacteria were below the levels of detection in subgingival plaque. We concluded that samples taken from subgingival plaque may be more useful for evaluating the proportion of periodontal bacteria in deep pockets than is the case for other samples. Therefore, for evaluation of periodontal bacteria, clinicians should consider the characteristics of the specimens obtained using different sampling methods.

Naturally fermented Jijelian black olives: microbiological characteristics and isolation of lactic acid bacteria

Directory of Open Access Journals (Sweden)

Karam, Nour-Eddine

2009-12-01

Full Text Available A study of the microflora of traditionally fermented black olives in Eastern Algeria is presented. A count of the following microbial groups was carried out: mesophilic bacteria, enterobacteria, lactic acid bacteria (LAB, staphylococci and yeast. In a second phase, the identification and assessment of the technological traits of LAB was performed. Seventeen lactic acid bacteria were isolated and identified. These isolates were represented by two genera: Lactobacillus and Leuconostoc. The results showed that Lactobacillus plantarum was the predominant species in this traditional product.Un estudio sobre la microflora de aceitunas negras fermentada por métodos tradicionales en el Este de Argelia es presentado. Se realizo el siguiente recuento de grupos de microorganismos: bacterias mesófilas, enterobacterias, bacterias ácido lácticas (LAB, staphylococcus y levaduras. En una segunda fase, la identificación y evaluación de aspectos tecnológicos de LAB fue realizada. Setenta bacterias ácido lácticas fueron aisladas e identificadas. Estos aislados contenían principalmente dos géneros: Lactobacillus y Leuconostoc. Los resultados mostraron que Lactobacillus plantarum fue la especie predominante en este producto tradicional.

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

Science.gov (United States)

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 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 Legionella control at

Microbial Count and Shelf Life of Phalsa(Grewia Asiatica) Juice

International Nuclear Information System (INIS)

Saddozai, A. A.; Mumtaz, A.; Raza, S.; Saleem, S. A.

2015-01-01

The study investigated the shelf life of laboratory developed phalsa juice at room temperature. Phalsa was purchased from local market, juice was prepared and kept in sterilized bottles at room temperature. Physicochemical and microbial and oragnoleptic quality of the juice was examined till two weeks. Microbial activity in phalsa juice increased while organoleptic attributes such as texture (mouth feel), flavour, taste, colour and overall acceptability of phalsa juice were decreased during the study period. The pH and TSS value were decreased from 3.99 to 3.54 and from 11.22 to 9.55, respectively after 2 weeks storage. Total plate count also showed decline from 6.2*10/sup -1/ to 3.2 * 10/sup -1/ cfuml/sup -1/ whereas yeast and mould counts increased simultaneously from 2.6*10/sup -1/ and nil to 5.5*10/sup -1/ and 2.4*10/sup -1/ cfuml/sup -1/, respectively during the storage. To increase shelf life of phalsa juice storage at refrigerated temperature with/without preservatives is recommended. (author)

Seasonal variations of nitrate reducing and denitrifying bacteria utilizing hexadecane in Mandovi estuary, Goa, West Coast of India

Digital Repository Service at National Institute of Oceanography (India)

Sousa, T.D.; Ingole, B.; Sousa, S.D.; Bhosle, S.

> cfu/ml on minimal media containing hexadecane as the sole carbon source. Highest bacterial counts were obtained during the monsoons. 22% of bacteria capable of hexadecane utilization were nitrate reducing and 12% were denitrifying. 29...

Identification of lead-resistant endophytic bacteria isolated from rice

International Nuclear Information System (INIS)

Perez-Cordero, Alexander; Barraza-Roman, Zafiro; Martinez-Pacheco, Dalila

2015-01-01

Resistance of endophytic bacteria in vitro was evaluated at different lead concentrations. The tissue samples of commercial rice varieties at tillering stage were collected during the first half of 2013, in Monteria, Cordoba, Colombia. Each tissue was subjected to surface cleaning. Endophytic bacteria were isolated in agar R_2A medium. The population density (CFU/g tissue) was determined from each tissue by direct counting of R_2A medium surface. Morphotypes were classified by shape, color, size and appearance. A total of 168 morphotypes were isolated from root, tillers and leaf of different commercial varieties of rice. The lead resistance test is performed in vitro, The lead resistance test was performed in vitro, by the suspensions of endophytic bacteria in log phase and inoculation in minimal medium with five concentrations of lead as Pb (NO_3)_2. The experiment was incubated at 32 degrees celsius and agitated at 150 rpm for five days. The measure of turbidimetry at 600 nm was conduced every hour afterstarting the test. Endophytic bacteria showed the ability to grow at concentrations of 100% of Pb as Pb (NO_3) _2. The presence of Burkholderia cepacia and Pseudomonas putida, which showed resistance to differents lead concentration was confirmed as result of the identification with kit API20E. (author) [es

Determining random counts in liquid scintillation counting

International Nuclear Information System (INIS)

Horrocks, D.L.

1979-01-01

During measurements involving coincidence counting techniques, errors can arise due to the detection of chance or random coincidences in the multiple detectors used. A method and the electronic circuits necessary are here described for eliminating this source of error in liquid scintillation detectors used in coincidence counting. (UK)

Antimicrobial copper alloys decreased bacteria on stethoscope surfaces.

Science.gov (United States)

Schmidt, Michael G; Tuuri, Rachel E; Dharsee, Arif; Attaway, Hubert H; Fairey, Sarah E; Borg, Keith T; Salgado, Cassandra D; Hirsch, Bruce E

2017-06-01

Stethoscopes may serve as vehicles for transmission of bacteria among patients. The aim of this study was to assess the efficacy of antimicrobial copper surfaces to reduce the bacterial concentration associated with stethoscope surfaces. A structured prospective trial involving 21 health care providers was conducted at a pediatric emergency division (ED) (n = 14) and an adult medical intensive care unit located in tertiary care facilities (n = 7). Four surfaces common to a stethoscope and a facsimile instrument fabricated from U.S. Environmental Protection Agency-registered antimicrobial copper alloys (AMCus) were assessed for total aerobic colony counts (ACCs), methicillin-resistant Staphylococcus aureus, gram-negative bacteria, and vancomycin-resistant enterococci for 90 days. The mean ACCs collectively recovered from all stethoscope surfaces fabricated from the AMCus were found to carry significantly lower concentrations of bacteria (pediatric ED, 11.7 vs 127.1 colony forming units [CFU]/cm 2 , P stethoscopes was the most heavily burdened surface; mean concentrations exceeded the health care-associated infection acquisition concentration (5 CFU/cm 2 ) by at least 25×, supporting that the stethoscope warrants consideration in plans mitigating microbial cross-transmission during patient care. Stethoscope surfaces fabricated with AMCus were consistently found to harbor fewer bacteria. Copyright © 2017 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

Effects of the gamma and ultrasound radiation in orange juice contaminated by Alicyclobacillus acidoterrestris;Efeitos das radiacoes gama e ultra-sonica em suco de laranja contaminado por Alicyclobacillus acidoterrestris

Energy Technology Data Exchange (ETDEWEB)

Pires, Cristiane Cassiolato, E-mail: cristiane_cassiolato@yahoo.com.b

2006-07-01

The orange juice has attributes could make that hard or even block the growth. of microorganisms, even so already have been isolated filamentous fungi, yeast, as well lactic and heat resistant bacteria. Alicyclobacillus acidoterrestris is a heat resistant bacteria that has done damage in the orange juice concentrated and frozen business. This bacterium is able the grown in temperatures below 35 deg C constituting a risk to orange juices that have been contaminated. Even those that already pass trough thermal treatment may deteriorated, if they are storage in place without refrigeration. As the traditional methods of decontamination and conservation have not been effective in unfeasible this bacterium, others methods of sterilization are need, among them are gamma and ultrasound radiation. The gamma radiation is able to sterilize foods and reduce the microbiology density, allowing in this way to enhance the period of storage. The ultrasound application has been used currently with the aim to microbiology control, been this technique effective in the microbial cell destruction. The Brazil, are the major producer and exporter of concentrated frozen orange juice, due to it, the present work was carried out to two experiments with the aim to determinate the resistance of the bacterium Alicyclobacillus acidoterrestris to gamma and ultrasound irradiation, in the orange juice, priory contaminated. The orange juice samples was diluted to 11,5 deg Brix and the bacteria suspensions was added, passed by process of radiation gamma with dose of 0, 2, 4, 6 and 8 kGy and ultrasonic with the frequency of 25, 35 and 42 kHz and with the time of exposition of 0, 1, 5, 10 and 20 minutes. After the treatment, the samples were stored in room temperature (25 +- 2 deg C) and of refrigeration (4 +- 1 deg C). For the microbial analyses, the samples were diluted in to decimal scale, plated by 'pour plate' technique. The Petri plates were storage in warn temperature (46 +- 2 deg C

Effects of the gamma and ultrasound radiation in orange juice contaminated by Alicyclobacillus acidoterrestris

International Nuclear Information System (INIS)

Pires, Cristiane Cassiolato

2006-01-01

The orange juice has attributes could make that hard or even block the growth. of microorganisms, even so already have been isolated filamentous fungi, yeast, as well lactic and heat resistant bacteria. Alicyclobacillus acidoterrestris is a heat resistant bacteria that has done damage in the orange juice concentrated and frozen business. This bacterium is able the grown in temperatures below 35 deg C constituting a risk to orange juices that have been contaminated. Even those that already pass trough thermal treatment may deteriorated, if they are storage in place without refrigeration. As the traditional methods of decontamination and conservation have not been effective in unfeasible this bacterium, others methods of sterilization are need, among them are gamma and ultrasound radiation. The gamma radiation is able to sterilize foods and reduce the microbiology density, allowing in this way to enhance the period of storage. The ultrasound application has been used currently with the aim to microbiology control, been this technique effective in the microbial cell destruction. The Brazil, are the major producer and exporter of concentrated frozen orange juice, due to it, the present work was carried out to two experiments with the aim to determinate the resistance of the bacterium Alicyclobacillus acidoterrestris to gamma and ultrasound irradiation, in the orange juice, priory contaminated. The orange juice samples was diluted to 11,5 deg Brix and the bacteria suspensions was added, passed by process of radiation gamma with dose of 0, 2, 4, 6 and 8 kGy and ultrasonic with the frequency of 25, 35 and 42 kHz and with the time of exposition of 0, 1, 5, 10 and 20 minutes. After the treatment, the samples were stored in room temperature (25 +- 2 deg C) and of refrigeration (4 +- 1 deg C). For the microbial analyses, the samples were diluted in to decimal scale, plated by 'pour plate' technique. The Petri plates were storage in warn temperature (46 +- 2 deg C) by 48 hours

Heterogeneous precipitation of silver nanoparticles on kaolinite plates

International Nuclear Information System (INIS)

Cabal, B; Moya, J S; Torrecillas, R; Malpartida, F

2010-01-01

Two different methods to obtain silver nanoparticles supported on kaolin crystals have been performed: the first one followed a thermal reduction and the second one a chemical reduction. In both cases, the silver nanoparticles with two different average particles size (ca.12 and 30 nm) were perfectly isolated and attached to the surface of the kaolin plates. The silver nanoparticles were localized mainly at the edge of the single crystal plates, the hydroxyl groups being the main centres of adsorption. The samples were fully characterized by XRD, UV-vis spectroscopy and TEM. The antimicrobial benefits of the composites were evaluated as antibacterial against common Gram-positive and Gram-negative bacteria, and antifungal activity against yeast. The results indicated a high antimicrobial activity for Escherichia coli JM 110 and Micrococcus luteus, while being inactive against yeast under our experimental conditions. The chemical analysis of Ag in the fermentation broths show that only a small portion of metal (<9 ppm) is released from the kaolin/metakaolin particles. Therefore, the risk of toxicity due to a high concentration of metal in the medium is minimized.

Heterogeneous precipitation of silver nanoparticles on kaolinite plates.

Science.gov (United States)

Cabal, B; Torrecillas, R; Malpartida, F; Moya, J S

2010-11-26

Two different methods to obtain silver nanoparticles supported on kaolin crystals have been performed: the first one followed a thermal reduction and the second one a chemical reduction. In both cases, the silver nanoparticles with two different average particles size (ca.12 and 30 nm) were perfectly isolated and attached to the surface of the kaolin plates. The silver nanoparticles were localized mainly at the edge of the single crystal plates, the hydroxyl groups being the main centres of adsorption. The samples were fully characterized by XRD, UV-vis spectroscopy and TEM. The antimicrobial benefits of the composites were evaluated as antibacterial against common Gram-positive and Gram-negative bacteria, and antifungal activity against yeast. The results indicated a high antimicrobial activity for Escherichia coli JM 110 and Micrococcus luteus, while being inactive against yeast under our experimental conditions. The chemical analysis of Ag in the fermentation broths show that only a small portion of metal (<9 ppm) is released from the kaolin/metakaolin particles. Therefore, the risk of toxicity due to a high concentration of metal in the medium is minimized.

Heterogeneous precipitation of silver nanoparticles on kaolinite plates

Energy Technology Data Exchange (ETDEWEB)

Cabal, B; Moya, J S [Instituto de Ciencia de Materiales de Madrid (ICMM-CSIC), 28049, Cantoblanco, Madrid (Spain); Torrecillas, R [Centro de Investigacion en Nanomateriales y NanotecnologIa (CINN), Consejo Superior de Investigaciones CientIficas (CSIC)-Universidad de Oviedo-UO-Principado de Asturias, Parque Tecnologico de Asturias, 33428, Llanera (Spain); Malpartida, F, E-mail: bcabal@icmm.csic.es [Centro Nacional de BiotecnologIa (CNB-CSIC), 28049, Cantoblanco, Madrid (Spain)

2010-11-26

Two different methods to obtain silver nanoparticles supported on kaolin crystals have been performed: the first one followed a thermal reduction and the second one a chemical reduction. In both cases, the silver nanoparticles with two different average particles size (ca.12 and 30 nm) were perfectly isolated and attached to the surface of the kaolin plates. The silver nanoparticles were localized mainly at the edge of the single crystal plates, the hydroxyl groups being the main centres of adsorption. The samples were fully characterized by XRD, UV-vis spectroscopy and TEM. The antimicrobial benefits of the composites were evaluated as antibacterial against common Gram-positive and Gram-negative bacteria, and antifungal activity against yeast. The results indicated a high antimicrobial activity for Escherichia coli JM 110 and Micrococcus luteus, while being inactive against yeast under our experimental conditions. The chemical analysis of Ag in the fermentation broths show that only a small portion of metal (<9 ppm) is released from the kaolin/metakaolin particles. Therefore, the risk of toxicity due to a high concentration of metal in the medium is minimized.

Significance of bacteria associated with invertebrates in drinking water distribution networks.

Science.gov (United States)

Wolmarans, E; du Preez, H H; de Wet, C M E; Venter, S N

2005-01-01

The implication of invertebrates found in drinking water distribution networks to public health is of concern to water utilities. Previous studies have shown that the bacteria associated with the invertebrates could be potentially pathogenic to humans. This study investigated the level and identity of bacteria commonly associated with invertebrates collected from the drinking water treatment systems as well as from the main pipelines leaving the treatment works. On all sampling occasions bacteria were isolated from the invertebrate samples collected. The highest bacterial counts were observed for the samples taken before filtration as was expected. There were, however, indications that optimal removal of invertebrates from water did not always occur. During the investigation, 116 colonies were sampled for further identification. The isolates represent several bacterial genera and species that are pathogenic or opportunistic pathogens of humans. Diarrhoea, meningitis, septicaemia and skin infections are among the diseases associated with these organisms. The estimated number of bacteria that could be associated with a single invertebrate (as based on average invertebrate numbers) could range from 10 to 4000 bacteria per organism. It can, therefore, be concluded that bacteria associated with invertebrates might under the worst case scenario pose a potential health risk to water users. In the light of the above findings it is clear that invertebrates in drinking water should be controlled at levels as low as technically and economically feasible.

Evaluation of Standard and Modified M-FC, MacConkey, and Teepol Media for Membrane Filtration Counting of Fecal Coliforms in Water

OpenAIRE

Grabow, W. O. K.; Hilner, C. A.; Coubrough, P.

1981-01-01

MacConkey agar, standard M-FC agar, M-FC agar without rosolic acid, M-FC agar with a resuscitation top layer, Teepol agar, and pads saturated with Teepol broth, were evaluated as growth media for membrane filtration counting of fecal coliform bacteria in water. In comparative tests on 312 samples of water from a wide variety of sources, including chlorinated effluents, M-FC agar without rosolic acid proved the medium of choice because it generally yielded the highest counts, was readily obtai...

Radiation counting statistics

Energy Technology Data Exchange (ETDEWEB)

Suh, M. Y.; Jee, K. Y.; Park, K. K.; Park, Y. J.; Kim, W. H

1999-08-01

This report is intended to describe the statistical methods necessary to design and conduct radiation counting experiments and evaluate the data from the experiment. The methods are described for the evaluation of the stability of a counting system and the estimation of the precision of counting data by application of probability distribution models. The methods for the determination of the uncertainty of the results calculated from the number of counts, as well as various statistical methods for the reduction of counting error are also described. (Author). 11 refs., 8 tabs., 8 figs.

Radiation counting statistics

Energy Technology Data Exchange (ETDEWEB)

Suh, M. Y.; Jee, K. Y.; Park, K. K. [Korea Atomic Energy Research Institute, Taejon (Korea)

1999-08-01

This report is intended to describe the statistical methods necessary to design and conduct radiation counting experiments and evaluate the data from the experiments. The methods are described for the evaluation of the stability of a counting system and the estimation of the precision of counting data by application of probability distribution models. The methods for the determination of the uncertainty of the results calculated from the number of counts, as well as various statistical methods for the reduction of counting error are also described. 11 refs., 6 figs., 8 tabs. (Author)

Radiation counting statistics

International Nuclear Information System (INIS)

Suh, M. Y.; Jee, K. Y.; Park, K. K.; Park, Y. J.; Kim, W. H.

1999-08-01

This report is intended to describe the statistical methods necessary to design and conduct radiation counting experiments and evaluate the data from the experiment. The methods are described for the evaluation of the stability of a counting system and the estimation of the precision of counting data by application of probability distribution models. The methods for the determination of the uncertainty of the results calculated from the number of counts, as well as various statistical methods for the reduction of counting error are also described. (Author). 11 refs., 8 tabs., 8 figs

Scale-down of vinegar production into microtiter plates using a custom-made lid.

Science.gov (United States)

Schlepütz, Tino; Büchs, Jochen

2014-04-01

As an important food preservative and condiment, vinegar is widely produced in industry by submerged acetic acid bacteria cultures. Although vinegar production is established on the large scale, up to now suitable microscale cultivation methods, e.g. using microtiter plates, are missing to enable high-throughput cultivation and to optimize fermentation conditions. In order to minimize evaporation losses of ethanol and acetic acid in a 48-well microtiter plate during vinegar production a new custom-made lid was developed. A diffusion model was used to calculate the dimensions of a hole in the lid to guarantee a suitable oxygen supply and level of ventilation. Reference fermentation was conducted in a 9-L bioreactor to enable the calculation of the proper cultivation conditions in the microtiter plate. The minimum dissolved oxygen tensions in the microtiter plate were between 7.5% and 23% of air saturation and in the same range as in the 9-L bioreactor. Evaporation losses of ethanol and acetic acid were less than 5% after 47 h and considerably reduced compared to those of microtiter plate fermentations with a conventional gas-permeable seal. Furthermore, cultivation times in the microtiter plate were with about 40 h as long as in the 9-L bioreactor. In conclusion, microtiter plate cultivations with the new custom-made lid provide a platform for high-throughput studies on vinegar production. Results are comparable to those in the 9-L bioreactor. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Chlorhexidine avoids skin bacteria recolonization more than triclosan.

Science.gov (United States)

Macias, Juan H; Alvarez, Mildred F; Arreguin, Virginia; Muñoz, Juan M; Macias, Alejandro E; Alvarez, Jose A

2016-12-01

We do not know whether differences exist between the residual effect of 2% chlorhexidine in 70% isopropyl alcohol when compared with 1% triclosan in 70% isopropyl alcohol. Using an analytic, longitudinal, controlled, and comparative experimental trial, with blinded measurements, we recruited healthy, adult volunteers from the University of Guanajuato who completed a stabilization phase of skin microbiota and had no history of skin allergies. Four 25-cm 2 areas of the inner surface of the forearms were designated for study: unscrubbed control for establishing baseline bacterial counts, scrubbed control with tridistilled water, scrubbed with chlorhexidine, and scrubbed with triclosan. Quantitative cultures were taken of all the areas at 0, 3, and 24 hours, using agar plates with neutralizing agents. A total of 135 healthy volunteers were tested. At 24 hours, the unscrubbed control counts were 288 CFU/cm 2 , whereas the scrubbed control counts were 96 CFU/cm 2 ; 24 CFU/cm 2 for chlorhexidine and 96 CFU/cm 2 for triclosan (Kruskal-Wallis χ 2 H = 64.27; P <.001). Chlorhexidine is the best antiseptic option when a prolonged antiseptic effect is needed; for instance, when implanting medical devices or performing surgical procedures. Copyright © 2016 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

An automated and highly efficient method for counting and measuring fluorescent foci in rod-shaped bacteria

DEFF Research Database (Denmark)

Nielsen, Henrik Jørck; Hansen, Flemming G.

2010-01-01

P>Direct measurements of cells from photo micrographs are becoming increasingly used when investigating the position and/or distribution of chromosomal loci in bacteria. In general, these measurements have been done manually, and without clear definition of how they are made. Here we present...

A new microtitre plate screening method for evaluating the viability of aerobic respiring bacteria in high surface biofilms.

Science.gov (United States)

Pérez, L M; Alvarez, B L; Codony, F; Fittipaldi, M; Adrados, B; Peñuela, G; Morató, J

2010-09-01

It is difficult to determine the effects of bactericidal compounds against bacteria in a biofilm because classical procedures for determining cell viability require several working days, multiple complicated steps and are frequently only applicable to cells in suspension. We attempt to develop a compact, inexpensive and versatile system to measure directly the extent of biofilm formation from water systems and to determine the viability of respiring bacteria in high surface biofilms. It has been reported that the reduction of tetrazolium sodium salts, such as XTT (sodium 3,3'-[1-[(phenylamino)carbonyl]-3,4-tetrazolium]Bis(4-methoxy)-6-nitro)benzene sulfonic acid hydrate), during active bacterial metabolism can be incorporated into a colorimetric method for quantifying cell viability. XTT is reduced to a soluble formazan compound during bacterial aerobic metabolism such that the amount of formazan generated is proportional to the bacterial biomass. We show here, for the first time, that this colorimetric approach can be used to determine the metabolic activity of adherent aerobic bacteria in a biofilm as a measure of cell viability. This technique has been used to estimate viability and proliferation of bacteria in suspension, but this is the first application to microbial communities in a real undisturbed biofilm. This simple new system can be used to evaluate the complex biofilm community without separating the bacteria from their support. Thus, the results obtained by this practice may be more representative of the circumstances in a natural system, opening the possibility to multiple potential applications.

Tower counts

Science.gov (United States)

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. 

Evaluation of inhibitory effects of iranian propolis against filamentous bacteria

International Nuclear Information System (INIS)

Eshraghi, S.; Valafar, S.

2008-01-01

To investigate the antibacterial activities of propolis in samples collected from Zanjan province Iran, against 25 pathogenic strains of bacteria. In order to evaluate the biological properties of methanol extract of propolis using agar distribution methods (disk and drop plate). Seven concentrations of methanolic extract of propolis were prepared and added drop wise to the bacterial seed layer cultured agar media individually. The diameter of the clear zone formed in each concentration was measured and correlated to the ability of the extracts to inhibit the growth of bacteria. Nocardia asteroides and N. brasiliensis has nearly shown the same susceptibility to various concentrations of propolis extract, and the complete clear zones revealed that this effect was quite remarkable. For other bacteria, different degrees of susceptibility to propolis were observed. We came to this conclusion that zones formed by 50mg/ml Amikacin in agar was similar to that of 5% concentration of propolis, and that the potency of propolis is 80% of Amikacin potency, which is the most effective antibiotic against Nocardia. (author)

Radiation-hard ceramic Resistive Plate Chambers for forward TOF and T0 systems

Energy Technology Data Exchange (ETDEWEB)

Akindinov, A., E-mail: Alexander.Akindinov@cern.ch [Institute for Theoretical and Experimental Physics, Moscow (Russian Federation); Dreyer, J.; Fan, X.; Kämpfer, B. [Helmholtz-Zentrum Dresden-Rossendorf, Dresden (Germany); Kiselev, S. [Institute for Theoretical and Experimental Physics, Moscow (Russian Federation); Kotte, R.; Garcia, A. Laso [Helmholtz-Zentrum Dresden-Rossendorf, Dresden (Germany); Malkevich, D. [Institute for Theoretical and Experimental Physics, Moscow (Russian Federation); Naumann, L. [Helmholtz-Zentrum Dresden-Rossendorf, Dresden (Germany); Nedosekin, A.; Plotnikov, V. [Institute for Theoretical and Experimental Physics, Moscow (Russian Federation); Stach, D. [Helmholtz-Zentrum Dresden-Rossendorf, Dresden (Germany); Sultanov, R.; Voloshin, K. [Institute for Theoretical and Experimental Physics, Moscow (Russian Federation)

2017-02-11

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.

Morphologic characterization and quantitative analysis on in vitro bacteria by nuclear techniques of measurement; Caracterizacao morfologica e analise quantitativa de bacterias in vitro por tecnicas nucleares de medidas

Energy Technology Data Exchange (ETDEWEB)

Lopes, Joana D' Arc Ramos

2001-10-01

The great difficulty to identify microorganisms (bacteria) from infectious processes is related to the necessary time to obtain a reliable result, about 72 hours. The purpose of this work is to establish a faster method to characterize bacterial morphologies through the use of neutron radiography, which can take about 5 hours. The samples containing the microorganisms, bacteria with different morphologies, after the appropriate microbiologic procedures were incubated with B{sup 10} for 30 minutes and soon after deposited in a plate of a solid detector of nuclear tracks (SSNTD), denominated CR-39. To obtain the images relative to bacteria, the detector was submitted to the flow of thermal neutrons of the order of 2.2 x 10{sup 5} n/cm{sup 2}.s from the J-9 channel of the Reactor Argonauta (IEN/CNEN). To observe the images from bacteria in each sample under an optical microscope, the sheets were chemically developed. The analysis of the images revealed morphologic differences among the genera (Gram positive from Gram-negative and coccus from bacillus), in samples containing either isolated or mixed bacteria. We thus verified the viability of the technique to achieve morphological characterization of different microorganisms. A quantitative approach seemed also to be feasible with the technique. The whole process took about 2 hours. (author)

Short communication: A comparison of biofilm development on stainless steel and modified-surface plate heat exchangers during a 17-h milk pasteurization run.

Science.gov (United States)

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

Bacteriological safety assessment, hygienic habits and cross-contamination risks in a Nigerian urban sample of household kitchen environment.

Science.gov (United States)

Ejechi, Bernard O; Ochei, Ono P

2017-06-01

Urban household kitchen environment was assessed for safety by determining their levels of indicator bacteria, hygienic habits and risk of cross-contamination. Household kitchens (60) were selected in Warri Town, Nigeria, by the multi-stage sampling technique. Contact surfaces, water and indoor kitchen air were analysed for aerobic plate counts, total and faecal coliforms using Nutrient and McConkey media by swab/rinse method, membrane filtration and sedimentation methods, respectively. Hygienic habits and risk of cross-contamination were assessed with structured questionnaire which included socio-demographic variables. On the basis of median counts, the prevalence of high counts (log cfu/cm 2 /m 3 /100 mL) of aerobic plate counts (>3.0), total coliforms (>1.0) and faecal coliforms (>0) on contact surfaces and air was high (58.0-92.0%), but low in water (30.0-40.0%). Pots, plates and cutleries were the contact surfaces with low counts. Prevalence of poor hygienic habits and high risk of cross-contamination was 38.6 and 67.5%, respectively. Education, occupation and kitchen type were associated with cross-contamination risk (P = 0.002-0.022), while only education was associated with hygienic habits (P = 0.03). Cross-contamination risk was related (P = 0.01-0.05) to aerobic plate counts (OR 2.30; CL 1.30-3.17), total coliforms (OR 5.63; CL 2.76-8.25) and faecal coliforms (OR 4.24; CL 2.87-6.24), while hygienic habit was not. It can be concluded that urban household kitchens in the Nigerian setting are vulnerable to pathogens likely to cause food-borne infections.

A novel microtiter plate radioimmunoassay of insulin autoantibody

International Nuclear Information System (INIS)

Huang Gan; Li Zhangwei; Jin Helai; Wang Xia; Wang Jianping; Zhou Zhiguang

2009-01-01

Objective: Insulin autoantibody (IAA) is known to exist in sera of type 1 diabetes mellitus (T1DM) patients and pre-T1DM individuals. The aim of this study was to establish a novel mierotiter plate radioimmunoassay (RIA) for IAA and evaluate its clinical value. Methods: Diluted 125 I-insulin was mixed with 5 μl serum samples in a 96-well microtiter plate and then incubated for 72 h on an orbital plate shaker (4 degree C). The immunocomplexes were transferred to another protein a coated Millipore plate, and then the plate was washed with Tri-Buffered Saline Tween-20 (TBT) buffer. Counts per minute (CPM) was measured with liquid scintillation and luminescence counter. The positive cut-off point of IAA index was defined as ≥0.06 based on the 99-percentile of the distribution in 317 healthy individuals. The specificity and sensitivity of the assay were calculated from the samples provided by the fourth Diabetes Autoantibodies Standardization Program (DASP 2005). The IAA levels were determined in 71 T1DM and 551 newly diagnosed type 2 diabetes (T2DM) patients, and 317 healthy controls. The t test, non-parametric test, χ 2 test and linear correlation analysis were performed on the data using SPSS 11.5 software. The concordance rate was estimated with Kappa value. Results: (1) The optimized testing condition was described as 2 x 10 4 CPM of 125 I-insulin, 5 μl serum sample and slowly horizontal shaking for 72 h. (2) The intra-assay CV was 4.8%-8.9% and inter-assay CV was 6.4%-10.5%. Based on DASP 2005 samples, the specificity and sensitivity of the assay were 97% (97/100) and 50% (25/50), respectively. Ninety-six serum samples with different IAA levels were selected and tested to compare between our new method and a domestic IAA RIA kit. The results showed that the IAA indices from the two methods were positively correlated (r= 0.678, P<0.001). The concordance rate was 72.9% (Kappa value=0.402). There were 25 samples with discordant results, which were positive for

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

Science.gov (United States)

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.

Irradiation control of pathogenic bacteria and their growth during storage time in cooled chicken

International Nuclear Information System (INIS)

Ha Yiming; Wang Feng; Fan Bei; Liu Shuliang; Ju Hua

2009-01-01

The growth of pathogenic bacteria during storage time and their D 10 values by irradiation in cooled chicken were evaluated. The total numbers of colony, E.coli 10003, Campylobacter jejuni33560 and CY04 of the D 10 values were 1.434 kGy, 0.408 kGy, 0.175 kGy, 0.2 kGy respectively in cooled chicken. The results show that total bacteria count in vacuum packaged cooled chicken sample is 5.66 lg(CFU/g) and 4.90 lg (CFU/g) after 3 kGy and 5 kGy irradiation. And in storage at 0∼4 degree C the storage shelf-life of irradiated vacuum packaged cooled chicken could extend to 21 d and 28 d. It can be deduced that pathogenic bacteria can be controlled effectively by irradiation. (authors)

Effects of gelling agent and extracellular signaling molecules on the culturability of marine bacteria

DEFF Research Database (Denmark)

Rygaard, Anita Mac; Schmidt Thøgersen, Mariane; Nielsen, Kristian Fog

2017-01-01

Only 1 % of marine bacteria are currently culturable using standard laboratory procedures and this is a major obstacle for our understanding of the biology of marine microorganisms and for the discovery of novel microbial natural products. Therefore, the purpose of the present study was to invest......Only 1 % of marine bacteria are currently culturable using standard laboratory procedures and this is a major obstacle for our understanding of the biology of marine microorganisms and for the discovery of novel microbial natural products. Therefore, the purpose of the present study...... was to investigate if improved cultivation conditions, including the use of an alternative gelling agent, and supplementation with signaling molecules, could improve the culturability of bacteria from seawater. Substituting agar with gellan gum improved viable counts 3 – 40-fold, depending on medium composition...

Radiation shielding plate

International Nuclear Information System (INIS)

Kobayashi, Torakichi; Sugawara, Takeo.

1983-01-01

Purpose: To reduce the weight and stabilize the configuration of a radiation shielding plate which is used in close contact with an object to be irradiated with radiation rays. Constitution: The radiation shielding plate comprises a substrate made of lead glass and a metallic lead coating on the surface of the substrate by means of plating, vapor deposition or the like. Apertures for permeating radiation rays are formed to the radiation shielding plate. Since the shielding plate is based on a lead glass plate, a sufficient mechanical strength can be obtained with a thinner structure as compared with the conventional plate made of metallic lead. Accordingly, if the shielding plate is disposed on a soft object to be irradiated with radiation rays, the object and the plate itself less deform to obtain a radiation irradiation pattern with distinct edges. (Moriyama, K.)

Plating on difficult-to-plate metals: what's new

International Nuclear Information System (INIS)

Wiesner, H.J.

1980-01-01

Some of the changes since 1970 in procedures for plating on such materials as titanium, molybdenum, silicon, aluminum, and gallium arsenide are summarized. While basic procedures for plating some of these materials were developed as many as 30 to 40 years ago, changes in the end uses of the plated products have necessitated new plating processes. In some cases, vacuum techniques - such as ion bombardment, ion implantation, and vacuum metallization - have been introduced to improve the adhesion of electrodeposits. In other cases, these techniques have been used to deposit materials upon which electrodeposits are required

Counting carbohydrates

Science.gov (United States)

Carb counting; Carbohydrate-controlled diet; Diabetic diet; Diabetes-counting carbohydrates ... Many foods contain carbohydrates (carbs), including: Fruit and fruit juice Cereal, bread, pasta, and rice Milk and milk products, soy milk Beans, legumes, ...

Effect of plate shapes in orifice plate type flowmeters

International Nuclear Information System (INIS)

Moeller, S.V.

1984-01-01

The study of unusual plate shapes in orifice plate type flowmeters is presented, with a view to providing data for the substitution of the plate with one centered circular orifice in those applications where its use is not possible. For this purpose, six pairs of plates with different forms, with and without chamfered edges, were made and tested in a closed water loop. Results show that, generally, the use of chamfers improves the results and, in the case of perforated and slotlike orificed plates, the narrow-ness of the fluid passage tends to make unnecessary its use. (Author) [pt

Plating laboratory

International Nuclear Information System (INIS)

Seamster, A.G.; Weitkamp, W.G.

1984-01-01

The lead plating of the prototype resonator has been conducted entirely in the plating laboratory at SUNY Stony Brook. Because of the considerable cost and inconvenience in transporting personnel and materials to and from Stony Brook, it is clearly impractical to plate all the resonators there. Furthermore, the high-beta resonator cannot be accommodated at Stony Brook without modifying the set up there. Consequently the authors are constructing a plating lab in-house

INHIBITION OF PATHOGENS BY SPOROGENIC BACTERIA ISOLATED FROM HONEY OF Melipona sp. (APIDAE: APINAE: MELIPONINI

Directory of Open Access Journals (Sweden)

KELY DAMIANA NOVAES DA SILVA

2016-01-01

Full Text Available The aim of this study was to isolate sporogenic bacteria from the honey of stingless bees Melipona sp., in dry forest, and to evaluate their antagonistic potential for medicinal employment purposes and animal production. The honey samples were collected in Serra Talhada - PE, where honey was taken from four different hives (in triplicate, totaling 12 samples. The samples were diluted and subjected to 80 ºC for 20 minutes to eliminate vegetative cells. The dilutions were plated onto nutrient agar and incubated at 30 ºC for 72 hours. Then the colony forming units (CFU were quantified. The samples were also plated onto malt agar and Sabouraud agar, and incubated at 30 ºC for 14 days for the growth of yeast and molds. Total and fecal coliforms were quantified by the most probable number method (MPN. Seven isolates (I of sporogenic bacteria ( Bacillus were obtained, however only four showed probiotic potential. Isolate I - 5 showed the greatest probiotic potential and inhibited the growth of Escherichia coli , Klebsiella sp., Pseudomonas aeruginosa, Salmonella sp., and Staphylococcus aureus . The growth of the Sarcina sp. was not inhibited by any isolate. No yeast, molds or coliforms were found. The Melipona sp. honey is a source of spore - forming bacteria and is antagonistic to microorganisms that contaminate honey. It has good microbiological quality.

COUNTS OF MICROORGANISMS CAUSING BOVINE MASTITIS AND STUDY OF ANTIMICROBIAL ACTION

Directory of Open Access Journals (Sweden)

Wanessa Oliveira Ribeiro

2014-02-01

Full Text Available Mastitis is an inflammation of the mammary gland caused mainly by microorganisms, altering the characteristics of milk and results in significant economic losses for this production complex. The study aimed to determine the main causative agents of bovine mastitis in a dairy farm in Rio Pomba city, Minas Gerais state, Brazil, and evaluate the use of plant extract and antibiotics commonly used in the control of microorganisms that cause this disease. Raw milk samples coming from 47 dairy cow were individually collected for microbiological evaluation. We also evaluated the sensitivity of isolates from the plant extract and the antibiotics commonly used in the farm. It was found that 17.0 %, 31.9 %, 85.4 % and 38.3 % of the samples presented, respectively, Staphylococcus aureus, coliforms, faecal coliforms and Escherichia coli. Furthermore, most of the samples showed counts of aerobic mesophilic microorganisms and Streptococcus sp. between 104 and 105 CFU.mL-1, while the counts of S. aureus ranged between 102 and 103 CFU.mL-1 in most of samples. A higher efficacy of tetracycline on the isolates of S. aureus was verified and of ampicillin on the E. coli isolates. All isolates of the latter bacteria were resistant to plant extract. Due to the high incidence of microorganisms, we emphasize the need for implementation of Good Agricultural Practices in milk production, because these bacteria are coming from hair, skin, mucous membranes of animals and/or belonging to the enteric microbiota of mammals, respectively.

Distribution of bacteria and fungi in the earthworm Libyodrillus violaceous (Annelida: Oligochaeta, a native earthworm from Nigeria

Directory of Open Access Journals (Sweden)

A. B Idowu

2006-03-01

Full Text Available Earthworms are soil invertebrates that play a key role in recycling organic matter in soils.In Nigeria, earthworms include Libyodrillus violaceous. Aerobic and anaerobic bacterial counts, as well as fungal counts of viable microorganisms in soils and gut sections, were made on twenty L. violaceous collected from different sites on the campus of the University of Agriculture, Abeokuta, Nigeria. The samples were collected between April and November, 2002. Numbers of microorganisms were higher in castings and gut sections than in uningested soil samples. The guts and their contents also had higher moisture and total nitrogen contents than the uningested soils. Bacteria and fungi isolated from the samples were identified by standard microbiological procedures on the bases of their morphological and biochemical characteristics. Isolated bacteria were identified as Staphylococcus, Bacillus spp., Pseudomonas aeruginosa, Streptococcus mutans, Clostridium, Spirocheata spp., Azotobacter spp., Micrococcus lylae, Acinetobacter spp., Halobacterium for bacteria. Yeast isolates were identified as Candida spp., Zygosaccharomyces spp., Pichia spp., and Saccharomyces spp while molds were identified as, Aspergillus spp., Pytium spp., Penicillium spp., Fusarium spp and Rhizopus spp. Of the five locations examined, the refuse dump area had the highest numbers of both aerobic and anaerobic organisms, followed by the arboretum while the cultivated land area recorded the lowest counts. The higher numbers of microorganisms observed in the gut sections and casts of the earthworms examined in this work reinforce the general concept that the gut and casts of earthworms show higher microbial diversity and activity than the surrounding soil. Rev. Biol. Trop. 54 (1: 49-58. Epub 2006 Mar 31.

The effect of laminar air flow and door openings on operating room contamination.

Science.gov (United States)

Smith, Eric B; Raphael, Ibrahim J; Maltenfort, Mitchell G; Honsawek, Sittisak; Dolan, Kyle; Younkins, Elizabeth A

2013-10-01

We evaluate the association of laminar airflow (LAF) and OR traffic with intraoperative contamination rates. Two sterile basins were placed in each room during 81 cases, one inside and one outside the LAF. One Replicate Organism Detection and Counting (RODAC) plate from each basin was sent for culture at successive 30-minute intervals from incision time until wound closure. At successive 30-minute intervals more plates were contaminated outside than inside the LAF. A negative binomial model showed that the bacteria colony forming units (CFU) depended on whether there were any door openings (P=0.02) and the presence of LAF (P=0.003). LAF decreases CFU by 36.6%. LAF independently reduces the risk of contamination and microbial counts for surgeries lasting 90 minutes or less. © 2013.

Honey characteristics after extraction and half-year storage

Directory of Open Access Journals (Sweden)

Vladimíra Kňazovická

2015-12-01

Full Text Available The aim of the study was to analyze the fresh honey after extracting and after half-year storage at room temperature. Overall, we analyzed 10 samples of rape (Brassica napus honey coming from district Vranov nad Toplou located in the eastern Slovakia. The analysis consisted of the evaluation of the physico-chemical parameters (water content, free acidity and electrical conductivity and microbiological evaluation (total plate count (TPC, counts of coliform bacteria, lactic acid bacteria, sporulating microorganisms and microscopic fungi. Water content, free acidity and electrical conductivity were measured according to IHC (2009, namely these parameters were detected by refractometer, titration and conductometer, respectively. We used dilution plating method for microbiological analysis. Fresh rape honey contained 18.3 ±1.0% of water. Free acidity of fresh rape honey was 12.7 ±2.0 meq.kg-1 and electrical conductivity was 0.14 mS.cm-1. After half a year of storage, water content and electrical conductivity decreased nonsignificantly and free acidity increased nonsignificantly. Stored honey samples meet the requirements of Decree 41/2012 and 106/2012. From microbiological point of view, fresh rape honey showed relatively high microbial counts. Mean values of TPC, sporulating microorganisms, lactic acid bacteria and yeasts exceeded 2.00 log cfu/g. All spotted microbial groups decreased in the stored honey comparing with the fresh honey. We found significant (p ˂0.01 differences of TPC, lactic acid bacteria and yeasts comparing the fresh and stored honey samples. Evaluating microbiological parameters, one sample of stored honey did not meet the requirements of Codex Alimentarius SR (2014. TPC exceeded the limit value. Based on the results we can conclude that all samples meet the requirements for good quality honey. Microbial counts in the honey decreased gradually. Probably, various microorganisms have important role in creation of the honey from

Detection of microbial concentration in ice-cream using the impedance technique.

Science.gov (United States)

Grossi, M; Lanzoni, M; Pompei, A; Lazzarini, R; Matteuzzi, D; Riccò, B

2008-06-15

The detection of microbial concentration, essential for safe and high quality food products, is traditionally made with the plate count technique, that is reliable, but also slow and not easily realized in the automatic form, as required for direct use in industrial machines. To this purpose, the method based on impedance measurements represents an attractive alternative since it can produce results in about 10h, instead of the 24-48h needed by standard plate counts and can be easily realized in automatic form. In this paper such a method has been experimentally studied in the case of ice-cream products. In particular, all main ice-cream compositions of real interest have been considered and no nutrient media has been used to dilute the samples. A measurement set-up has been realized using benchtop instruments for impedance measurements on samples whose bacteria concentration was independently measured by means of standard plate counts. The obtained results clearly indicate that impedance measurement represents a feasible and reliable technique to detect total microbial concentration in ice-cream, suitable to be implemented as an embedded system for industrial machines.

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

Science.gov (United States)

Vahid Dastjerdi, Elahe; Abdolazimi, Zahra; Ghazanfarian, Marzieh; Amdjadi, Parisa; Kamalinejad, Mohammad; Mahboubi, Arash

2014-12-01

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. 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. 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. 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.

Drought resistant of bacteria producing exopolysaccharide and IAA in rhizosphere of soybean plant (Glycine max) in Wonogiri Regency Central Java Indonesia

Science.gov (United States)

Susilowati, A.; Puspita, A. A.; Yunus, A.

2018-03-01

Drought is one of the main problem which limitating the agriculture productivity in most arid region such as in district Eromoko, Wuryantro and SelogiriWonogiri Central Java Indonesia. Bacteria are able to survive under stress condition by producte exopolysaccharide. This study aims to determine the presence of exopolysaccharide-producing drought-resistant bacteria on rhizosphere of soybean (Glycine max) and to determine the species of bacteria based on 16S rRNA gene. Isolation of bacteria carried out by the spread plate method. The decreased of osmotic potential for screening drought tolerant bacteria according to the previous equation [12]. Selection of exopolysaccharide-producing bacteria on solid media ATCC 14 followed by staining the capsule. 16S rRNA gene amplification performed by PCR using primers of 63f and 1387r. The identificationof the bacteria is determined by comparing the results of DNA sequence similarity with bacteria databank in NCBI database. The results showed 11 isolates were exopolysaccharide-producing drought tolerant bacteria. The identity of the bacteria which found are Bacillus sp, Bacillus licheniformis, Bacillus megaterium and Bacillus pumilus.

Isolation of naphthalene-degrading bacteria from tropical marine sediments

International Nuclear Information System (INIS)

Zhuang, W.-Q.; Tay, J.-H.; Maszenan, A.M.; Tay, S.T.-L.

2003-01-01

Oil pollution is a major environmental concern in many countries, and this has led to a concerted effort in studying the feasibility of using oil-degrading bacteria for bioremediation. Although many oil-degrading bacteria have been isolated from different environments, environmental conditions can impose a selection pressure on the types of bacteria that can reside in a particular environment. This study reports the successful isolation of two indigenous naphthalene-degrading bacteria from oil-contaminated tropical marine sediments by enrichment culture. Strains MN-005 and MN-006 were characterized using an extensive range of biochemical tests. The 16S ribosomal deoxyribonucleic acid (rDNA) sequence analysis was also performed for the two strains. Their naphthalene degradation capabilities were determined using gas chromatography and DAPI counting of bacterial cells. Strains MN-005 and MN-006 are phenotypically and phylogenetically different from each other, and belong to the genera Staphylococcus and Micrococcus, respectively. Strains MN-005 and MN-006 has maximal specific growth rates (μ max ) of 0.082±0.008 and 0.30±0.02 per hour, respectively, and half-saturation constants (K s ) of 0.79±0.10 and 2.52±0.32 mg per litre, respectively. These physiological and growth studies are useful in assessing the potential of these indigenous isolates for in situ or ex situ naphthalene pollutant bioremediation in tropical marine environments. (author)

Identification of lead- resistant endophytic bacteria isolated from rice.

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Alexander Pérez-Cordero

2015-06-01

Full Text Available   The objective of this study was to evaluate in vitro the endophytic bacteria resistance to different lead concentrations. The sampling was undertaken in the first half of 2013, when tissue samples of commercial varieties of rice at tillering stage were collected in Montería, Cordoba, Colombia. Each tissue was subjected to surface cleaning. Endophytic bacteria in agar R2A medium were isolated. Population density (CFU/g tissue was determined from each tissue, by direct counting of R2A medium surface. morphotypes were classified by shape, color, size, and appearance. A total of 168 morphotypes were isolated from root, tillers, and leaf of different commercial varieties of rice. The lead resistance test was performed in vitro, to do that, suspensions of endophytic bacteria in log phase were prepared and inoculated in minimal medium with five concentrations of lead as Pb(NO32. The experiment was incubated at 32 °C and agitated at 150 rpm, for five days. Every hour afterstarting the test, turbidimetry measuring at 600 nm was conducted. Results showed the ability of endophytic bacteria to grow at concentrations of 100% of Pb as Pb(NO32. The results of the identification with kit API20E confirmed the presence of Burkholderia cepacia and Pseudomonas putida, which showed resistance to different lead concentrations.

Comparative study on disinfection potency of spore forming bacteria by electron-beam irradiation and gamma-ray irradiation

International Nuclear Information System (INIS)

Takizawa, Hironobu; Suzuki, Satoru; Suzuki, Tetsuya; Takama, Kozo; Hayashi, Toru; Yasumoto, Kyoden.

1990-01-01

Along with gamma-ray irradiation, electron-beam irradiation (EB) is a method to disinfect microorganisms which cause food decomposition and food-poisoning. The present study was undertaken to compare sterilization efficacy of EB and gamma-ray irradiation on bacterial spores and vegetative cells under various conditions. Spores of Bacillus pumilus, a marker strain for irradiation study, and Bacillus stearothermophilus known as a thermophilic bacteria were irradiated by electron-beam and gamma-ray separately at irradiation dose of 0 to 10 kGy on combination of wet/dry and aerobic/anaerobic conditions. Sterilization effect of irradiation on spores was evaluated by colony counting on agar plates. Results showed that both EB and gamma-ray irradiation gave sufficient sterilization effect on spores, and the sterilization effect increased exponentially with irradiation dose. The sterilization effect of gamma-ray irradiation was higher than that of EB in all cases. Higher disinfection effect was observed under aerobic condition. The present study suggests that oxygen supply in EB is more important than gamma-ray irradiation. No results suggesting that chlorine ion at 0.1 ppm (as available chlorine concentration) enhanced the sterilization efficacy of either EB or gamma-ray irradiation was obtained under any conditions examined. (author)

Comparative study on disinfection potency of spore forming bacteria by electron-beam irradiation and gamma-ray irradiation

Energy Technology Data Exchange (ETDEWEB)

Takizawa, Hironobu; Suzuki, Satoru; Suzuki, Tetsuya; Takama, Kozo [Hokkaido Univ., Hakodate (Japan). Faculty of Fisheries; Hayashi, Toru; Yasumoto, Kyoden

1990-10-01

Along with gamma-ray irradiation, electron-beam irradiation (EB) is a method to disinfect microorganisms which cause food decomposition and food-poisoning. The present study was undertaken to compare sterilization efficacy of EB and gamma-ray irradiation on bacterial spores and vegetative cells under various conditions. Spores of Bacillus pumilus, a marker strain for irradiation study, and Bacillus stearothermophilus known as a thermophilic bacteria were irradiated by electron-beam and gamma-ray separately at irradiation dose of 0 to 10 kGy on combination of wet/dry and aerobic/anaerobic conditions. Sterilization effect of irradiation on spores was evaluated by colony counting on agar plates. Results showed that both EB and gamma-ray irradiation gave sufficient sterilization effect on spores, and the sterilization effect increased exponentially with irradiation dose. The sterilization effect of gamma-ray irradiation was higher than that of EB in all cases. Higher disinfection effect was observed under aerobic condition. The present study suggests that oxygen supply in EB is more important than gamma-ray irradiation. No results suggesting that chlorine ion at 0.1 ppm (as available chlorine concentration) enhanced the sterilization efficacy of either EB or gamma-ray irradiation was obtained under any conditions examined. (author).

Contamination pathways of spore-forming bacteria in a vegetable cannery.

Science.gov (United States)

Durand, Loïc; Planchon, Stella; Guinebretiere, Marie-Hélène; André, Stéphane; Carlin, Frédéric; Remize, Fabienne

2015-06-02

Spoilage of low-acid canned food during prolonged storage at high temperatures is caused by heat resistant thermophilic spores of strict or facultative bacteria. Here, we performed a bacterial survey over two consecutive years on the processing line of a French company manufacturing canned mixed green peas and carrots. In total, 341 samples were collected, including raw vegetables, green peas and carrots at different steps of processing, cover brine, and process environment samples. Thermophilic and highly-heat-resistant thermophilic spores growing anaerobically were counted. During vegetable preparation, anaerobic spore counts were significantly decreased, and tended to remain unchanged further downstream in the process. Large variation of spore levels in products immediately before the sterilization process could be explained by occasionally high spore levels on surfaces and in debris of vegetable combined with long residence times in conditions suitable for growth and sporulation. Vegetable processing was also associated with an increase in the prevalence of highly-heat-resistant species, probably due to cross-contamination of peas via blanching water. Geobacillus stearothermophilus M13-PCR genotypic profiling on 112 isolates determined 23 profile-types and confirmed process-driven cross-contamination. Taken together, these findings clarify the scheme of contamination pathway by thermophilic spore-forming bacteria in a vegetable cannery. Copyright © 2015 Elsevier B.V. All rights reserved.

Create Your Plate

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Full Text Available ... Plate Share Create Your Plate ! Share: Seven Simple Steps to Create Your Plate It's simple and effective ... foods within each food category. Try these seven steps to get started: Using your dinner plate, put ...

A combination of direct viable count and fluorescence in situ hybridization for specific enumeration of viable Lactobacillus delbrueckii subsp.bulgaricus and Streptococcus thermophilus.

Science.gov (United States)

García-Hernández, J; Moreno, Y; Amorocho, C M; Hernández, M

2012-03-01

We have developed a direct viable count (DVC)-FISH procedure for quickly and easily discriminating between viable and nonviable cells of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains, the traditional yogurt bacteria. direct viable count method has been modified and adapted for Lact. delbrueckii subsp. bulgaricus and Strep. thermophilus analysis by testing different times of incubation and concentrations of DNA-gyrase inhibitors. DVC procedure has been combined with fluorescent in situ hybridization (FISH) for the specific detection of viable cells of both bacteria with specific rRNA oligonucleotide probes (DVC-FISH). Of the four antibiotics tested (novobiocin, nalidixic acid, pipemidic acid and ciprofloxacin), novobiocin was the most effective for DVC method and the optimum incubation time was 7 h for both bacteria. The number of viable cells was obtained by the enumeration of specific hybridized cells that were elongated at least twice their original length for Lactobacillus and twice their original size for Streptococcus. This technique was successfully applied to detect viable cells in inoculated faeces. Results showed that this DVC-FISH procedure is a quick and culture-independent useful method to specifically detect viable Lact. delbrueckii subsp. bulgaricus and Strep. thermophilus in different samples, being applied for the first time to lactic acid bacteria. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

Biodiversity of polycyclic aromatic hydrocarbon-degrading bacteria from deep sea sediments of the Middle Atlantic Ridge.

Science.gov (United States)

Cui, Zhisong; Lai, Qiliang; Dong, Chunming; Shao, Zongze

2008-08-01

The bacteria involved in the biodegradation of polycyclic aromatic hydrocarbons (PAHs) in deep sea subsurface environments are largely unknown. In order to reveal their biodiversity, sediments from 2.2 m under the bottom surface at a water depth of 3542 m were sampled on the Middle Atlantic Ridge with a gravity column sampler. The sediments were promptly enriched with either crude oil or a mixture of PAHs (naphthalene, phenanthrene and pyrene) as the sole carbon source, and further enriched with the PAH mixture mentioned above in the lab. The resulting consortia were named C2CO and C2PPN respectively. Their bacterial composition was analysed with plate cultivation, PCR-DGGE and 16S rDNA library analysis. On plates, isolates belonging to Pseudoalteromonas, Halomonas, Marinobacter, Thalassospira and Tistrella dominated the culturable populations. With PCR-DGGE, five major bands closely related to Cycloclasticus, Alteromonas, Thalassospira, Alcanivorax and Rhodospirillaceae were detected in consortium C2CO, while only one major band of Cycloclasticus was detected in consortium C2PPN. In addition, the dynamics of community structure in response to aromatic substrate alterations were examined. As a result, three ribotypes of Cycloclasticus were detected by 16S rDNA library analysis, one which played a key role in phenanthrene degradation; two Alteromonas bacteria dominated the naphthalene reselected consortium. Although bacteria of the two genera grew as the main members of the communities, none of them were isolated, probably owing to their poor cultivability. These results confirm that bacteria of Cycloclasticus are important obligate PAH degraders in marine environments, and coexist with other degrading bacteria that inhabit the deep subsurface sediment of the Atlantic. This supports the view that PAH accumulation and bioattenuation occur in remote areas consistently and continuously.

Development of a nematode offspring counting assay for rapid and simple soil toxicity assessment.

Science.gov (United States)

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.

Evaluation of biogenic amines and microbial counts throughout the ripening of goat cheeses from pasteurized and raw milk.

Science.gov (United States)

Novella-Rodríguez, Sonia; Veciana-Nogués, M Teresa; Roig-Sagués, Artur X; Trujillo-Mesa, Antonio J; Vidal-Carou, M Carmen

2004-05-01

The effect of the hygienic quality of milk on changes in microbial counts and biogenic amine content was evaluated during ripening of goat cheeses manufactured from pasteurized and raw milks at 1, 14, 30, 60 and 90 d. The original milk, rennet, curd and whey were also included in the study. The pH, salt content and extent of proteolysis in the cheese were also evaluated. Spermidine and spermine were the main amines in raw milk, while they were minor amines in cheeses. Other amines increased markedly during ripening, tyramine being the main amine in cheese made from raw milk and cadaverine and putrescine in those produced from pasteurized milk. Enterobacteriaceae counts decreased during ripening whereas those of lactic acid bacteria increased, especially lactobacilli and enterococci. Cheese made from raw milk showed higher microbial counts during ripening than those made from pasteurized milk, especially for Enterobacteriaceae and enterococci, counts being 2 or 3 log units higher. Raw milk cheese showed remarkably higher biogenic amines compared with pasteurized milk cheeses. Therefore, pasteurization of milk causes a decrease in final biogenic amine content of cheese as a result of the reduction of its microbial counts.

Whey protein isolate/cellulose nanofibre/TiO2 nanoparticle/rosemary essential oil nanocomposite film: Its effect on microbial and sensory quality of lamb meat and growth of common foodborne pathogenic bacteria during refrigeration.

Science.gov (United States)

Alizadeh Sani, Mahmood; Ehsani, Ali; Hashemi, Mohammad

2017-06-19

The use of biodegradable nanocomposite films in active packaging is of great importance since they can have a controlled release of antimicrobial compounds. This study was conducted to evaluate the efficacy of whey protein isolate (WPI)/cellulose nanofibre (CNF) nanocomposite films containing 1.0% (w/w) titanium dioxide (TiO 2 ) and 2.0% (w/v) rosemary essential oil (REO) in preserving the microbial and sensory quality of lamb meat during the storage at 4±1°C. Initially, the best concentration of each compound to be added to the film was determined by micro-dilution and disc diffusion methods. The microbial and sensory properties of lamb meat were controlled in two groups (control and treatment) over 15days of storage. Then, the samples were analysed for total viable count (TVC), Pseudomonas spp. count, Enterobacteriaceae count, Lactic acid bacteria (LAB) count, inoculated Staphylococcus aureus count, Listeria monocytogenes count, and Escherichia coli O 157 :H 7 count. Microbial analysis and nine-point hedonic scale was applied for the sensory analysis. Results indicated that the use of nanocomposite films significantly reduced the bacterial counts of treatment group. Higher inhibition effect was observed on Gram-positive bacteria than on Gram-negative bacteria (Psensory evaluations also showed that the use of nanocomposite films significantly increased the shelf life of treated meat (15days) compared to the control meat (6days). Based on the results of this study, the edible nanocomposite films were effective in preserving the microbial and sensory qualities of lamb meat; therefore, this application is recommended in meat especially red meat. Copyright © 2017 Elsevier B.V. All rights reserved.

Impact of feathers and feather follicles on broiler carcass bacteria.

Science.gov (United States)

Cason, J A; Hinton, A; Buhr, R J

2004-08-01

Genetically featherless and feathered broiler siblings were used to test the contribution of feathers and feather follicles to the numbers of aerobic bacteria, Escherichia coli, and Campylobacter in whole-carcass rinse samples taken immediately after carcasses were defeathered for 30 or 60 s. Numbers of spoilage bacteria were counted after the same fully processed carcasses were stored for 1 wk at 2 degrees C. In each of 3 replications, twenty-eight 11-wk-old, mixed-sex, genetically featherless or feathered broilers were processed in a laboratory processing facility. Immediately after individual defeathering in a mechanical picker, carcasses were sampled using a carcass rinse technique. Carcasses were eviscerated, immersion chilled at 2 degrees C for 30 min, individually bagged, and stored for 1 wk at 2 degrees C, after which all carcasses were rinsed again, and spoilage bacteria in the rinsate were enumerated. There were no significant differences (P defeathering and no differences between carcasses picked for 30 or 60 s. There were no differences in numbers of spoilage bacteria after 1 wk of refrigeration for any of the feather presence-picking length combinations. Although the defeathering step in poultry processing has been identified as an opportunity for bacterial contamination from the intestinal tract and cross-contamination between carcasses, the presence of feathers and feather follicles does not make a significant difference in carcass bacterial contamination immediately after defeathering or in spoilage bacteria after 1 wk of refrigeration.

Microbiological quality of drinking water from dispensers in roadside restaurants of Bangladesh.

Science.gov (United States)

Moniruzzaman, M; Akter, S; Islam, M A; Mia, Z

2011-01-15

The microbiological status of water from dispensers in different roadside restaurants of Dhaka city and Savar area was analyzed in this study. Seven samples from Dhaka and 8 samples of Savar were checked. The heterotrophic plate count was in a range of 1.0 x 10(3) CFU mL(-1) to 2.0 x 10(4) CFU mL(-1) (from new bottles), 1.0 x 10(3) to 1.5 x 10(4) CFU mL(-1) (after dispensation), and 1.5 x 10(3) CFU mL(-1) to 1.0 x l0(5) CFU mL(-1) (from serving glass). In several of the samples, the heterotrophic plate count was higher than the count in water from new bottle or after dispensation, suggesting added contamination from the serving glass. 80% of the samples were contaminated with total and fecal coliform bacteria, which render these waters unacceptable for human consumption. The samples were found to contain gram negative bacteria like E coli, Shigella sp., Klebsiella sp., Enterobacter sp., Pseudomonas sp., and Salmonella sp., which are potential pathogens and thus pose a serious threat to public health. This study elucidates the importance of monitoring the bottling companies and the restaurants and put them under strict regulations to prevent future outbreak of any water borne diseases caused by consumption of dispensed water.

[Comparison of techniques for coliform bacteria extraction from sediment of Xochimilco Lake, Mexico].

Science.gov (United States)

Fernández-Rendón, Carlos L; Barrera-Escorcia, Guadalupe

2013-01-01

The need to separate bacteria from sediment in order to appropriately count them has led to test the efficacy of different techniques. In this research, traditional techniques such as manual shaking, homogenization, ultrasonication, and surfactant are compared. Moreover, the possibility of using a set of enzymes (pancreatine) and an antibiotic (ampicillin) for sediment coliform extraction is proposed. Samples were obtained from Xochimilco Lake in Mexico City. The most probable number of coliform bacteria was determined after applying the appropriate separation procedure. Most of the techniques tested led to numbers similar to those of the control (manual shaking). Only with the use of ampicillin, a greater total coliform concentration was observed (Mann-Whitney, z = 2.09; p = 0.03). It is possible to propose the use of ampicillin as a technique for total coliform extraction; however, it is necessary to consider sensitivity of bacteria to the antibiotic.

Standardization of 241Am by digital coincidence counting, liquid scintillation counting and defined solid angle counting

International Nuclear Information System (INIS)

Balpardo, C.; Capoulat, M.E.; Rodrigues, D.; Arenillas, P.

2010-01-01

The nuclide 241 Am decays by alpha emission to 237 Np. Most of the decays (84.6%) populate the excited level of 237 Np with energy of 59.54 keV. Digital coincidence counting was applied to standardize a solution of 241 Am by alpha-gamma coincidence counting with efficiency extrapolation. Electronic discrimination was implemented with a pressurized proportional counter and the results were compared with two other independent techniques: Liquid scintillation counting using the logical sum of double coincidences in a TDCR array and defined solid angle counting taking into account activity inhomogeneity in the active deposit. The results show consistency between the three methods within a limit of a 0.3%. An ampoule of this solution will be sent to the International Reference System (SIR) during 2009. Uncertainties were analysed and compared in detail for the three applied methods.

Shelf life of pasteurized microfiltered milk containing 2% fat.

Science.gov (United States)

Caplan, Z; Barbano, D M

2013-01-01

The goal of this research was to produce homogenized milk containing 2% fat with a refrigerated shelf life of 60 to 90 d using minimum high temperature, short time (HTST) pasteurization in combination with other nonthermal processes. Raw skim milk was microfiltered (MF) using a Tetra Alcross MFS-7 pilot plant (Tetra Pak International SA, Pully, Switzerland) equipped with Membralox ceramic membranes (1.4 μm and surface area of 2.31 m(2); Pall Corp., East Hills, NY). The unpasteurized MF skim permeate and each of 3 different cream sources were blended together to achieve three 2% fat milks. Each milk was homogenized (first stage: 17 MPa, second stage: 3 MPa) and HTST pasteurized (73.8°C for 15s). The pasteurized MF skim permeate and the 3 pasteurized homogenized 2% fat milks (made from different fat sources) were stored at 1.7 and 5.7°C and the standard plate count for each milk was determined weekly over 90 d. When the standard plate count was >20,000 cfu/mL, it was considered the end of shelf life for the purpose of this study. Across 4 replicates, a 4.13 log reduction in bacteria was achieved by MF, and a further 0.53 log reduction was achieved by the combination of MF with HTST pasteurization (73.8°C for 15s), resulting in a 4.66 log reduction in bacteria for the combined process. No containers of MF skim milk that was pasteurized after MF exceeded 20,000 cfu/mL bacteria count during 90 d of storage at 5.7°C. The 3 different approaches used to reduce the initial bacteria and spore count of each cream source used to make the 2% fat milks did not produce any shelf-life advantage over using cold separated raw cream when starting with excellent quality raw whole milk (i.e., low bacteria count). The combination of MF with HTST pasteurization (73.8°C for 15s), combined with filling and packaging that was protected from microbial contamination, achieved a refrigerated shelf life of 60 to 90 d at both 1.7 and 5.7°C for 2% fat milks. Copyright © 2013 American

Benjamin Thompson, Count Rumford Count Rumford on the nature of heat

CERN Document Server

Brown, Sanborn C

1967-01-01

Men of Physics: Benjamin Thompson - Count Rumford: Count Rumford on the Nature of Heat covers the significant contributions of Count Rumford in the fields of physics. Count Rumford was born with the name Benjamin Thompson on March 23, 1753, in Woburn, Massachusetts. This book is composed of two parts encompassing 11 chapters, and begins with a presentation of Benjamin Thompson's biography and his interest in physics, particularly as an advocate of an """"anti-caloric"""" theory of heat. The subsequent chapters are devoted to his many discoveries that profoundly affected the physical thought

Paper microzone plates.

Science.gov (United States)

Carrilho, Emanuel; Phillips, Scott T; Vella, Sarah J; Martinez, Andres W; Whitesides, George M

2009-08-01

This paper describes 96- and 384-microzone plates fabricated in paper as alternatives to conventional multiwell plates fabricated in molded polymers. Paper-based plates are functionally related to plastic well plates, but they offer new capabilities. For example, paper-microzone plates are thin (approximately 180 microm), require small volumes of sample (5 microL per zone), and can be manufactured from inexpensive materials ($0.05 per plate). The paper-based plates are fabricated by patterning sheets of paper, using photolithography, into hydrophilic zones surrounded by hydrophobic polymeric barriers. This photolithography used an inexpensive formulation photoresist that allows rapid (approximately 15 min) prototyping of paper-based plates. These plates are compatible with conventional microplate readers for quantitative absorbance and fluorescence measurements. The limit of detection per zone loaded for fluorescence was 125 fmol for fluorescein isothiocyanate-labeled bovine serum albumin, and this level corresponds to 0.02 the quantity of analyte per well used to achieve comparable signal-to-noise in a 96-well plastic plate (using a solution of 25 nM labeled protein). The limits of detection for absorbance on paper was approximately 50 pmol per zone for both Coomassie Brilliant Blue and Amaranth dyes; these values were 0.4 that required for the plastic plate. Demonstration of quantitative colorimetric correlations using a scanner or camera to image the zones and to measure the intensity of color, makes it possible to conduct assays without a microplate reader.

Isolation, Biochemical and Molecular Identification, and In-Vitro Antimicrobial Resistance Patterns of Bacteria Isolated from Bubaline Subclinical Mastitis in South India.

Directory of Open Access Journals (Sweden)

P L Preethirani

Full Text Available Buffaloes are the second largest source of milk. Mastitis is a major impediment for milk production, but not much information is available about bubaline mastitis, especially subclinical mastitis. The aim of this study was to (a investigate the application of various tests for the diagnosis of bubaline subclinical mastitis, (b identify the major bacteria associated with it, and (c evaluate the antibiotic resistance pattern of the bacteria. To this end, 190 quarter milk samples were collected from 57 domesticated dairy buffaloes from organized (64 samples and unorganized (126 samples sectors. Of these, 48.4%, 40.0%, 45.8%, 61.1%, and 61.6% were positive for subclinical mastitis by somatic cell count, electrical conductivity, California mastitis test, bromothymol blue test, and N-acetyl glucosaminidase test, respectively. As compared to the gold standard of somatic cell count, California mastitis test performed the best. However, a combination of the two methods was found to be the best option. Microbiological evaluation, both by biochemical methods as well as by monoplex and multiplex polymerase chain reaction, revealed that coagulase-negative staphylococci were the most predominant (64.8% bacteria, followed by streptococci (18.1%, Escherichia coli (9.8% and Staphylococcus aureus (7.3%. Most of the pathogens were resistant to multiple antibiotics, especially to β-lactam antibiotics. We propose that California mastitis test be combined with somatic cell count for diagnosis of subclinical mastitis in domestic dairy buffaloes. Further, our results reveal high resistance of the associated bacteria to the β-lactam class of antibiotics, and a possible major role of coagulase-negative staphylococci in causing the disease in India.

Isolation, Biochemical and Molecular Identification, and In-Vitro Antimicrobial Resistance Patterns of Bacteria Isolated from Bubaline Subclinical Mastitis in South India.

Science.gov (United States)

Preethirani, P L; Isloor, Shrikrishna; Sundareshan, S; Nuthanalakshmi, V; Deepthikiran, K; Sinha, Akhauri Y; Rathnamma, D; Nithin Prabhu, K; Sharada, R; Mukkur, Trilochan K; Hegde, Nagendra R

2015-01-01

Buffaloes are the second largest source of milk. Mastitis is a major impediment for milk production, but not much information is available about bubaline mastitis, especially subclinical mastitis. The aim of this study was to (a) investigate the application of various tests for the diagnosis of bubaline subclinical mastitis, (b) identify the major bacteria associated with it, and (c) evaluate the antibiotic resistance pattern of the bacteria. To this end, 190 quarter milk samples were collected from 57 domesticated dairy buffaloes from organized (64 samples) and unorganized (126 samples) sectors. Of these, 48.4%, 40.0%, 45.8%, 61.1%, and 61.6% were positive for subclinical mastitis by somatic cell count, electrical conductivity, California mastitis test, bromothymol blue test, and N-acetyl glucosaminidase test, respectively. As compared to the gold standard of somatic cell count, California mastitis test performed the best. However, a combination of the two methods was found to be the best option. Microbiological evaluation, both by biochemical methods as well as by monoplex and multiplex polymerase chain reaction, revealed that coagulase-negative staphylococci were the most predominant (64.8%) bacteria, followed by streptococci (18.1%), Escherichia coli (9.8%) and Staphylococcus aureus (7.3%). Most of the pathogens were resistant to multiple antibiotics, especially to β-lactam antibiotics. We propose that California mastitis test be combined with somatic cell count for diagnosis of subclinical mastitis in domestic dairy buffaloes. Further, our results reveal high resistance of the associated bacteria to the β-lactam class of antibiotics, and a possible major role of coagulase-negative staphylococci in causing the disease in India.

Growth and persistence of pathogens on granular activated carbon filters.

Science.gov (United States)

Camper, A K; LeChevallier, M W; Broadaway, S C; McFeters, G A

1985-01-01

Three enteric pathogens Yersinia enterocolitica O:8, Salmonella typhimurium, and enterotoxigenic Escherichia coli, were examined for their ability to colonize granular activated carbon (GAC) in pure cultures and in the presence of autochthonous river water organisms. All three organisms readily colonized sterile GAC and maintained populations of ca. 10(5) to 10(7) CFU g-1 for 14 days when suspended in sterile river water. Exposure of pathogen biofilms on GAC to unsterile river water resulted in a gradual decline in pathogens on the carbon (0.08 to 0.14 log day-1). When pathogens were introduced to sterile GAC in the presence of heterotrophic plate count organisms, they attached at levels similar to those in the pure cultures and then decreased (0.10 to 0.22 log day-1). When added with heterotrophic plate count bacteria to GAC supporting a mature biofilm of native river water bacteria, they attached at a lower level (1.0 X 10(4) to 4.6 X 10(4) CFU g-1) and decreased at a more rapid rate (0.11 to 0.70 log day-1). PMID:3911903

Molecular identification and technological characterization of lactic acid bacteria isolated from fermented kidney beans flours (Phaseolus vulgaris L. and P. coccineus) in northwestern Argentina.

Science.gov (United States)

Sáez, Gabriel D; Hébert, Elvira M; Saavedra, Lucila; Zárate, Gabriela

2017-12-01

Legumes are an important protein source in developing countries and their flours represent an attractive alternative for the manufacture of gluten free products. In the present study, 4 kidney bean varieties (Alubia, Pallar, Black and Red beans) commonly cultivated in northwestern Argentina, were milled and spontaneously fermented in order to isolate and select autochthonous lactic acid bacteria (LAB) with relevant technological and functional properties for usage as starter cultures. Twelve doughs were fermented with daily back-slopping at 37°C for 6days and evolution of total mesophiles, lactic acid bacteria, and yeasts and molds populations were followed by plate counting. A combination of phenotypic and genotypic methods including (GTG) 5 -based PCR fingerprinting and 16S rRNA gene sequencing were used to differentiate and identify the isolated LAB to species level. LAB counts ranged from around 0.89±0.81 to 8.74±0.03logcfu/g with a pH decline from 6.4 to 3.9 throughout fermentation. Four genera and nine species of LAB: Enterococcus durans, E. faecium, E. mundtii, E. casseliflavus; Lactobacillus rhamnosus, Lactococcus garvieae, Weissella cibaria and W. paramesenteroides were found on kidney beans. Twenty five LAB strains were assessed for their abilities to grow on kidney bean extracts, acidifying capacities (pH and acidification rates), amylolytic, proteolytic, tannase and gallate decarboxylase activities as well as pathogens inhibition by antimicrobials. Based on these properties E. durans CRL 2178 and W. paramesenteroides CRL 2182 were inoculated singly and combined in Alubia kidney bean flour and fermented for 24h at 37°C. LAB strains were beneficial for removing trypsin inhibitors and tannins from sourdoughs and for improving amino acids and phenolics contents, increasing the antioxidant activities of kidney bean matrices. Selected strains have potential as starter cultures for obtaining fermented bean products with high nutritional and functional

Symbiotic interaction of endophytic bacteria with arbuscular mycorrhizal fungi and its antagonistic effect on Ganoderma boninense.

Science.gov (United States)

Sundram, Shamala; Meon, Sariah; Seman, Idris Abu; Othman, Radziah

2011-08-01

Endophytic bacteria (Pseudomonas aeruginosa UPMP3 and Burkholderia cepacia UMPB3), isolated from within roots of oil palm (Elaeis guineensis Jacq.) were tested for their presymbiotic effects on two arbuscular mcorrhizal fungi, Glomus intraradices UT126 and Glomus clarum BR152B). These endophytic bacteria were also tested for antagonistic effects on Ganoderma boninense PER 71, a white wood rot fungal pathogen that causes a serious disease in oil palm. Spore germination and hyphal length of each arbuscular mycorrhizal fungal (AMF) pairing with endophytic bacteria was found to be significantly higher than spores plated in the absence of bacteria. Scanning electron microscopy (SEM) showed that the endophytic bacteria were scattered, resting or embedded on the surface hyaline layer or on the degraded walls of AMF spores, possibly feeding on the outer hyaline spore wall. The antagonistic effect of the endophytic bacteria was expressed as severe morphological abnormalities in the hyphal structures of G. boninense PER 71. The effects of the endophytic bacteria on G. boninense PER 71 hyphal structures were observed clearly under SEM. Severe inter-twisting, distortion, lysis and shriveling of the hyphal structures were observed. This study found that the effect of endophytic bacteria on G. intraradices UT126 and G. clarum BR152B resembled that of a mycorrhiza helper bacteria (MHB) association because the association significantly promoted AMF spore germination and hyphal length. However, the endophytic bacteria were extremely damaging to G. boninense PER 71.

Epifluorescent direct counts of bacteria and viruses from topsoil of various desert dust storm regions

Science.gov (United States)

Gonzalez-Martin, Cristina; Teigell-Perez, Nuria; Lyles, Mark; Valladares, Basilio; Griffin, Dale W.

2013-01-01

Topsoil from arid regions is the main source of dust clouds that move through the earth's atmosphere, and microbial communities within these soils can survive long-range dispersion. Microbial abundance and chemical composition were analyzed in topsoil from various desert regions. Statistical analyses showed that microbial direct counts were strongly positively correlated with calcium concentrations and negatively correlated with silicon concentrations. While variance between deserts was expected, it was interesting to note differences between sample sites within a given desert region, illustrating the 'patchy' nature of microbial communities in desert environments.

Counting cormorants

DEFF Research Database (Denmark)

Bregnballe, Thomas; Carss, David N; Lorentsen, Svein-Håkon

2013-01-01

This chapter focuses on Cormorant population counts for both summer (i.e. breeding) and winter (i.e. migration, winter roosts) seasons. It also explains differences in the data collected from undertaking ‘day’ versus ‘roost’ counts, gives some definitions of the term ‘numbers’, and presents two...

Interactions between stream fungi and bacteria associated with decomposing leaf litter at different levels of nutrient availability

Science.gov (United States)

Vladislav Gulis; Keller Suberkropp

2003-01-01

We examined the potential for interactions between aquatic hyphomycetes and bacteria isolated from leaves decaying in a headwater stream. In agar plate assays, culture filtrates of each of 28 aquatic hyphomycete isolates tested (5 species) inhibited bacterial growth (16 Gram-negative bacterial isolates belonging to 6 colony morphotypes were tested). Inhibition of...

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

Science.gov (United States)

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 (pbiofilm 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 (pbiofilms 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 the biofilm

Isolation and life cycle characterization of lytic viruses infecting heterotrophic bacteria and cyanobacteria

DEFF Research Database (Denmark)

Middelboe, Mathias; Chan, Amy; Bertelsen, Sif Koldborg

2010-01-01

Basic knowledge on viruses infecting heterotrophic bacteria and cyanobacteria is key to future progress in understanding the role of viruses in aquatic systems and the influence of virus–host interactions on microbial mortality, biogeochemical cycles, and genetic exchange. Such studies require......, and discusses the applications and limitations of different isolation procedures. Most work on phage isolation has been carried out with aerobic heterotrophic bacteria and cyanobacteria, culturable both on agar plates and in enriched liquid cultures. The procedures presented here are limited to lytic viruses...... infecting such hosts. In addition to the isolation procedures, methods for life cycle characterization (one-step growth experiments) of bacteriophages and cyanophages are described. Finally, limitations and drawbacks of the proposed methods are assessed and discussed...

Selection of the Chrome Reduction Bacteria in the Waste of Tanning Leather Industries by Ozonization Method

International Nuclear Information System (INIS)

Yazid, M.; Aris Bastianudin; Widdi Usada

2007-01-01

Selection of the chrome reduction bacteria in the waste of tanning leather industries by ozonization method has been done. The objectives of this research was to obtain isolate bacteria from the waste with chrome contain, so that expected can be used for chrome bioremediation agent for arrange to improved the waste treatment for tanning leather industries. Selection of bacteria in the waste was carried out by ozonization method with time variation 0 to 210 minutes by time interval 15 minutes. Isolation bacteria was carried out was grown on the BHI media for 24 hours at 37°C temperature. So be inoculated by streak plate method on the TBX, MC, EA, CTM and BP media. Characterization of bacteria was done by saw the colonies morphology, sel morphology and biochemical characterization. So, identification of isolate bacteria by matching profile method. The result of this research can be obtained 5 isolate bacteria BCR1, BCR2, BCR3, BCR4 and BCR5 with the different phenotypic character. From the five isolate can be selected resistance ozon isolate until 180 minutes time ozonization were BCR 2, were identified belong to the genus of Bacillus. The examination results showed that the isolate bacteria be able to reduction of the chrome concentration in the waste of tanning leather industries by 71.03 %. Efficiency. (author)

Distinctive colonization of Bacillus sp. bacteria and the influence of the bacterial biofilm on electrochemical behaviors of aluminum coatings.

Science.gov (United States)

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.

Comparative evaluation of apical extrusion of bacteria using hand and rotary systems : An in vitro study

Science.gov (United States)

Ghivari, Sheetal B; Kubasad, Girish C; Deshpande, Preethi

2012-01-01

Aim: To evaluate the bacteria extruded apically during root canal preparation using two hand and rotary instrumentation techniques. Materials and Methods: Eighty freshly extracted mandibular premolars were mounted in bacteria collection apparatus. Root canals were contaminated with the pure culture of Enterococcus fecalis (ATCC 29212) and dried at 37°C for 24 h. Bacteria extruded were collected, incubated in brain heart infusion agar for 24 h at 36°C and the colony forming units (CFU) were counted. Statistical Analysis: The mean number of colony forming units were calculated by One-way ANOVA and comparison between the groups made by multiple comparison (Dunnet D) test. Results: The step-back technique extruded highest number of bacteria in comparison to other hand and rotary Ni–Ti systems. Conclusion: Under the limitation of this study all hand and rotary instrumentation techniques extruded bacteria. Among all the instrumentation techniques step-back technique extruded more number of bacteria and K-3 system the least. Further in vivo research in this direction could provide more insight into the biologic factors associated and focus on bacterial species that essentially play a major role in post instrumentation flare-ups. PMID:22368332

Locking screw-plate interface stability in carbon-fibre reinforced polyetheretherketone proximal humerus plates.

Science.gov (United States)

Hak, David J; Fader, Ryan; Baldini, Todd; Chadayammuri, Vivek B S

2017-09-01

Carbon-fibre reinforced polyetheretherketone (CFR-PEEK) plates have recently been introduced for proximal humerus fracture treatment. The purpose of this study was to compare the locking screw-plate interface stability in CFR-PEEK versus stainless steel (SS) proximal humerus plates. Locking screw mechanical stability was evaluated independently in proximal and shaft plate holes. Stiffness and load to failure were tested for three conditions: (1) on-axis locking screw insertion in CFR-PEEK versus SS plates, (2) on-axis locking screw insertion, removal, and reinsertion in CFR-PEEK plates, and (3) 10-degree off-axis locking screw insertion in CFR-PEEK plates. Cantilever bending at a rate of 1 mm/minute was produced by an Instron machine and load-displacement data recorded. Shaft locking screw load to failure was significantly greater in CFR-PEEK plates compared to SS plates (746.4 ± 89.7 N versus 596.5 ± 32.6 N, p PEEK plates (p PEEK plates. The mechanical stability of locking screws in CFR-PEEK plates is comparable or superior to locking screws in SS plates.

An experimental study of the attachment of bacteria to submerged surfaces in marine environment

International Nuclear Information System (INIS)

Fera, Ph.

1985-09-01

The seasonal variations of the bacterial settling of three materials (stainless steel, aluminium, polycarbonate filters) have been studied inside an open system of circulating seawater (0.7 m.s -1 ). The fixed bacteria counting have been carried out by scanning electron microscopy and epi-fluorescence microscopy. From the results of the first part of this work, it appears that the growth kinetics of the microbial bio-film, and the densities of the bacteria fixed after 15 days of immersion are higher during summer. Qualitatively, the composition of the number of fixed bacteria evolve with immersion time and with the season. The continuous injection of 0.1 ppm of chlorine in the seawater feeding the experimental system, seems not to be sufficient to prevent, for a long time, the settling of a great number of bacteria. The second part of this work deals with the experimental study of the settling of an aluminium surface by a pseudomonas, isolated of the seawater and submitted or not to conditions of preliminary fast. (O.M.)

Petroleum residues degradation in laboratory-scale by rhizosphere bacteria isolated from the mangrove ecosystem

Science.gov (United States)

Rinanti, A.; Nainggolan, I. J.

2018-01-01

This research is about petroleum bioremediation experiment to obtain bacterial isolate from mangrove ecosystem which potentially degrade petroleum. It was conducted in an Erlenmeyer batch system filled with growth medium of Stone Mineral Salt Solution (SMSS) plus petroleum residue, placed in an incubator shaker with a rotation speed of 120 rpm, temperature 3000C, for 14 research days. Indigenous bacteria that have been isolated and identified from the roots of mangrove plants are Ochrobactrum anthropi and Bacillus sp., Ralstonia pickettii and Bacillus circulans. Those bacteriain both monoculture and consortium form (mixed culture) are incorporated into erlenmeyer as remediator agents. All bacteria can utilize hydrocarbon compounds, but Ralstonia pickettii and Bacillus circulans reached exponential phase faster with more cell count than other bacteria. Compared to single cultures, petroleum degradation by a bacterial consortium provides a higher TPH reduction efficiency, i.e. at 5%, 10%, and 15% of initial TPH of 94.4%, 72%, and 80.3%, respectively. This study proved that all bacteria could optimize hydrocarbon compounds up to 15% TPH load.

Voltage-current characteristics of a pin-plate system with different plate configurations

International Nuclear Information System (INIS)

Feng, Zhuangbo; Long, Zhengwei

2013-01-01

In this paper, the voltage-current (V-I) characteristics of a pin-plate system with four types of collection plate configurations are studied experimentally. The collection plates consider a single metal plate, a metal plate with a fly ash cake layer, a metal plate with a clean filter media and a metal plate with a dirty filter media. The results show that the clean filter media has no obvious effect on the V-I characteristics. But the dirty filter media reduces the current density because of its high resistance. The thick fly ash cake layer increase current density because of the anti-corona effect but the increment is not very obvious.

Kinetics of killing Listeria monocytogenes by macrophages: correlation of 3H-DNA release from labeled bacteria and changes in numbers of viable organisms by mathematical model

International Nuclear Information System (INIS)

Davies, W.A.

1982-01-01

Conventional methods of assessing antibacterial activities of macrophages by viable counting are limited by the precision of the statistics and are difficult to interpret quantitatively because of unrestrained extracellular growth of bacteria. An alternative technique based on the release of radioactive DNA from labeled bacteria has been offered as overcoming these drawbacks. To assess it for use with macrophages I have made a correlation with the conventional viable counting method using a mathematical model. Opsonized Listeria monocytogenes labeled with 3 H-thymidine were exposed to rat macrophages for periods up to 4 hr. Numbers of viable bacteria determined after sonication increased exponentially in the absence of live cells and this growth rate was progressively inhibited by increasing numbers of macrophages. After a lag period of 30-60 min soluble 3 H appeared in the supernatant, the amount increasing with time and numbers of macrophages. To correlate these data I developed a mathematical model that considered that changes in numbers of viable organisms were due to the difference between rates of 1) growth of extracellular bacteria and 2) killing within the macrophage. On the basis of this model curves of best fit to the viable counts data were used to predict the release of radioactivity, assuming that death of a bacterium led to the total release of its label. These predictions and the experimental data agreed well, the lag period of 30-60 min between death of the bacterium and release of radioactivity being consistent with intracellular digestion. Release of soluble radioactivity appears to be an accurate reflection of the number of bacteria killed within the macrophage

Radiation induced damage to the lipid contents of bacteria and cultured mammalian cells

International Nuclear Information System (INIS)

Gholipour Khalili, K.

1993-01-01

In this study, exponentially growing phase of E. Coli. K12-N167 and cultured mouse leukemic L5178Y were used to study the effect of gamma irradiation on phospholipid contents. Following irradiation, both bacteria and cultured cells were incubated with either 14 C or 32 P labelled precursors for periods of cell division time. Phospholipid composition and their contents were detected in both the bacteria and cultured cells by using liquid scintillation counting and autoradiography methods. In contrast, as radiation dose increased, the Phospholipid contents were decreased in the both bacteria and cultured cells. It was concluded that the changes of phospholipid contents may result to altered activities of phospholipid pathway enzymes damaged by a radiation dose. The results of this investigation would be helpful in control of induced radiation damages in cell killings in radiation workers and radiation treatment of human cancer in the clinics. (author). 35 refs, 3 figs, 4 tabs

Evaluation of bean and soy tempeh influence on intestinal bacteria and estimation of antibacterial properties of bean tempeh.

Science.gov (United States)

Kuligowski, Maciej; Jasińska-Kuligowska, Iwona; Nowak, Jacek

2013-01-01

In this study the effect of bean tempeh on the growth of Bacillus subtilis, Escherichia coli, Lactobacillus acidophilus and Lactobacillus paracasei bacteria was investigated. Antibacterial activity was observed only in relation to the bacteria Bacillus subtilis. The effect of tempeh products on human intestinal microflora was also assessed. Bean and soy tempeh were culinarily processed and next digested in conditions simulating the human digestive tract (one of the digestive tracts was equipped with a mechanism simulating absorption). Soy tempeh stimulated most the growth of bacteria of the genus Bifidobacterium, while bean tempeh that of Escherichia coli. Using simulation of absorption for the digestion of fried soy tempeh resulted in a higher rise in the bacteria count of the genus Lactobacillus, while after digestion of fried bean tempeh the highest increase was recorded for Bifidobacterium and E. coli.

Effect of Associated Bacteria on the Growth and Toxicity of Alexandrium catenella

Science.gov (United States)

Uribe, Paulina; Espejo, Romilio T.

2003-01-01

Saprophytic bacteria in cultures of the marine dinoflagellate Alexandrium catenella were removed to assess their effect on growth and paralytic shellfish poisoning toxin production of this dinoflagellate. The actual axenic status was demonstrated by the lack of observable bacteria both immediately after treatment and following extended incubation in the absence of antibiotics. Bacteria were measured by counting CFU and also by epifluorescence microscopy and PCR amplification of bacterial 16S-23S spacer ribosomal DNA to detect noncultivable bacteria. Removal of bacteria did not have any effect on the growth of the dinoflagellate except for the inhibition of A. catenella disintegration after reaching the stationary phase. Toxicity was determined in dinoflagellate cell extracts by different methods: high-performance liquid chromatography (HPLC); an electrophysiological test called the Electrotest, which measures the inhibition of saxitoxin-sensitive Na+ channels expressed in a cell line; and a mouse bioassay, which measures the toxic effect on the whole mammal neuromuscular system. A lower toxicity of the dinoflagellates in axenic culture was observed by these three methods, though the difference was significant only by the mouse bioassay and HPLC methods. Altogether the results indicate that axenic cultures of A. catenella are able to produce toxin, though the total toxicity is probably diminished to about one-fifth of that in nonaxenic cultures. PMID:12514056

Morphologic characterization and quantitative analysis on in vitro bacteria by nuclear techniques of measurement

International Nuclear Information System (INIS)

Lopes, Joana D'Arc Ramos

2001-10-01

The great difficulty to identify microorganisms (bacteria) from infectious processes is related to the necessary time to obtain a reliable result, about 72 hours. The purpose of this work is to establish a faster method to characterize bacterial morphologies through the use of neutron radiography, which can take about 5 hours. The samples containing the microorganisms, bacteria with different morphologies, after the appropriate microbiologic procedures were incubated with B 10 for 30 minutes and soon after deposited in a plate of a solid detector of nuclear tracks (SSNTD), denominated CR-39. To obtain the images relative to bacteria, the detector was submitted to the flow of thermal neutrons of the order of 2.2 x 10 5 n/cm 2 .s from the J-9 channel of the Reactor Argonauta (IEN/CNEN). To observe the images from bacteria in each sample under an optical microscope, the sheets were chemically developed. The analysis of the images revealed morphologic differences among the genera (Gram positive from Gram-negative and coccus from bacillus), in samples containing either isolated or mixed bacteria. We thus verified the viability of the technique to achieve morphological characterization of different microorganisms. A quantitative approach seemed also to be feasible with the technique. The whole process took about 2 hours. (author)