Uncertainty of quantitative microbiological methods of pharmaceutical analysis.

PubMed

Gunar, O V; Sakhno, N G

2015-12-30

The total uncertainty of quantitative microbiological methods, used in pharmaceutical analysis, consists of several components. The analysis of the most important sources of the quantitative microbiological methods variability demonstrated no effect of culture media and plate-count techniques in the estimation of microbial count while the highly significant effect of other factors (type of microorganism, pharmaceutical product and individual reading and interpreting errors) was established. The most appropriate method of statistical analysis of such data was ANOVA which enabled not only the effect of individual factors to be estimated but also their interactions. Considering all the elements of uncertainty and combining them mathematically the combined relative uncertainty of the test results was estimated both for method of quantitative examination of non-sterile pharmaceuticals and microbial count technique without any product. These data did not exceed 35%, appropriated for a traditional plate count methods. Copyright © 2015 Elsevier B.V. All rights reserved.

A generalized plate method for estimating total aerobic microbial count.

PubMed

Ho, Kai Fai

2004-01-01

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

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

PubMed Central

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

1974-01-01

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

Rapid enumeration of viable bacteria by image analysis

NASA Technical Reports Server (NTRS)

Singh, A.; Pyle, B. H.; McFeters, G. A.

1989-01-01

A direct viable counting method for enumerating viable bacteria was modified and made compatible with image analysis. A comparison was made between viable cell counts determined by the spread plate method and direct viable counts obtained using epifluorescence microscopy either manually or by automatic image analysis. Cultures of Escherichia coli, Salmonella typhimurium, Vibrio cholerae, Yersinia enterocolitica and Pseudomonas aeruginosa were incubated at 35 degrees C in a dilute nutrient medium containing nalidixic acid. Filtered samples were stained for epifluorescence microscopy and analysed manually as well as by image analysis. Cells enlarged after incubation were considered viable. The viable cell counts determined using image analysis were higher than those obtained by either the direct manual count of viable cells or spread plate methods. The volume of sample filtered or the number of cells in the original sample did not influence the efficiency of the method. However, the optimal concentration of nalidixic acid (2.5-20 micrograms ml-1) and length of incubation (4-8 h) varied with the culture tested. The results of this study showed that under optimal conditions, the modification of the direct viable count method in combination with image analysis microscopy provided an efficient and quantitative technique for counting viable bacteria in a short time.

Semiquantitative determination of mesophilic, aerobic microorganisms in cocoa products using the Soleris NF-TVC method.

PubMed

Montei, Carolyn; McDougal, Susan; Mozola, Mark; Rice, Jennifer

2014-01-01

The Soleris Non-fermenting Total Viable Count method was previously validated for a wide variety of food products, including cocoa powder. A matrix extension study was conducted to validate the method for use with cocoa butter and cocoa liquor. Test samples included naturally contaminated cocoa liquor and cocoa butter inoculated with natural microbial flora derived from cocoa liquor. A probability of detection statistical model was used to compare Soleris results at multiple test thresholds (dilutions) with aerobic plate counts determined using the AOAC Official Method 966.23 dilution plating method. Results of the two methods were not statistically different at any dilution level in any of the three trials conducted. The Soleris method offers the advantage of results within 24 h, compared to the 48 h required by standard dilution plating methods.

SURVIVAL OF SALMONELLA SPECIES IN RIVER WATER.

EPA Science Inventory

The survival of four Salmonella strains in river water microcosms was monitored using culturing techniques, direct counts, whole cell hybridization, scanning electron microscopy, and resuscitation techniques via the direct viable count method and flow cytrometry. Plate counts of...

SURVIVAL OF SALMONELLA SPECIES IN RIVER WATER

EPA Science Inventory

The survival of four Salmonella strains in river water microcosms was monitored by culturing techniques, direct counts, whole-cell hybridization, scanning electron microscopy, and resuscitation techniques via the direct viable count method and flow cytometry. Plate counts of bact...

Total and Viable Legionella pneumophila Cells in Hot and Natural Waters as Measured by Immunofluorescence-Based Assays and Solid-Phase Cytometry ▿†

PubMed Central

Parthuisot, N.; Binet, M.; Touron-Bodilis, A.; Pougnard, C.; Lebaron, P.; Baudart, J.

2011-01-01

A new method was developed for the rapid and sensitive detection of viable Legionella pneumophila. The method combines specific immunofluorescence (IF) staining using monoclonal antibodies with a bacterial viability marker (ChemChrome V6 cellular esterase activity marker) by means of solid-phase cytometry (SPC). IF methods were applied to the detection and enumeration of both the total and viable L. pneumophila cells in water samples. The sensitivity of the IF methods coupled to SPC was 34 cells liter−1, and the reproducibility was good, with the coefficient of variation generally falling below 30%. IF methods were applied to the enumeration of total and viable L. pneumophila cells in 46 domestic hot water samples as well as in cooling tower water and natural water samples, such as thermal spring water and freshwater samples. Comparison with standard plate counts showed that (i) the total direct counts were always higher than the plate counts and (ii) the viable counts were higher than or close to the plate counts. With domestic hot waters, when the IF assay was combined with the viability test, SPC detected up to 3.4 × 103 viable but nonculturable L. pneumophila cells per liter. These direct IF methods could be a powerful tool for high-frequency monitoring of domestic hot waters or for investigating the occurrence of viable L. pneumophila in both man-made water systems and environmental water samples. PMID:21742913

Evaluation of propidium monoazide real-time PCR for enumeration of probiotic lactobacilli microencapsulated in calcium alginate beads.

PubMed

Oketič, K; Matijašić, B Bogovič; Obermajer, T; Radulović, Z; Lević, S; Mirković, N; Nedović, V

2015-01-01

The aim of the study was to evaluate real-time PCR coupled with propidium monoazide (PMA) treatment for enumeration of microencapsulated probiotic lactobacilli microencapsulated in calcium alginate beads. Lactobacillus gasseri K7 (CCM 7710) and Lactobacillus delbrueckii subsp. bulgaricus (CCM 7712) were analysed by plate counting and PMA real-time PCR during storage at 4 °C for 90 days. PMA was effective in preventing PCR amplification of the target sequences of DNA released from heat-compromised bacteria. The values obtained by real-time PCR of non-treated samples were in general higher than those obtained by real-time PCR of PMA-treated samples or by plate counting, indicating the presence of sub-lethally injured cells. This study shows that plate count could not be completely replaced by culture independent method PMA real-time PCR for enumeration of probiotics, but may rather complement the well-established plate counting, providing useful information about the ratio of compromised bacteria in the samples.

Rapid surface colony counts determination with three new miniaturised techniques.

PubMed

Malik, K A

1977-01-01

Three different miniaturised methods for the rapid surface viable counting are described. The methods were tried in parallel to seven different existing methods (Table 1) for viable counts and were found to be easier, quicker and insome cases more accurate. The techniques require about 10% of the material and time needed for conventional spread-plates method and the results were in no way inferior to that (Table 1 and 2). Mini agar discs were cut aseptically with an especially designed stainless steel agar disc cutter (25 mm internal and 28 mm external diameter, Fig. 1b) or with a test tube of similar diameter. The area of the resulted mini-agar-disc of 25 mm diameter was kept such (about 1/10th of the normal plate) that the ratio of the colony-bearing area to the inoculm remained the same as on big plates in spread-plate-method (Table 2). In normal Petri dishes (about 90 mm diameter) up to seven mini agar discs were possible to cut. Each small agar disc was seperated from the other mini-disc by a distance of at least 6 mm (Fig. 1a). The empty place around the disc was still enlarged during over drying of the plates and during incubation. This created complete isolation from the neighbouring disc. For micro-determination of surface viable counts 10 micronl from each dilution was delivered on a well-dired mini-disc with a piston micropipette. The inoculm was immediately spread on the whole mini-disc with a specially designed flame sterilizable platinum-Mini-spreader (Fig. 2a). No spinning of the plate was needed. Alternatively the dropping pipette and spreader was replaced by a calibrated platinum wire Loop-spreader (Fig. 2b). A loop of 3 mm internal diameter made from a platinum-iridium wire of 0.75 mm thickness proved most useful and carried a drop of 10 micronl. Differences especially in surface tension of various diluting fluids did not influence to drop of this size and no recalibration was needed for water and nutrient broth. The loop was further shaped to Loop-spreader form. From each bacterial suspension 10 micronl were carried and spread on each mini-disc. The method is useful for pathogenic organisms as the loop can readily be flame sterilized. For routine purposes where only approximate numbers of bacteria need to be known a still rapid semiquantitative method was deviced making use of a calibrated stainless steel Stamping-disc (Fig. 2c). A disc of 25mm diameter and 1 mm thickness delivered approximateyl 10 microlitres of supensions and was found to be most useful to stamp seven dilutions on a single plate. In collections and bacteriology laboratories where by conventional methods large number of plates are to be plated and counted the presented techniques could prove most convenient, rapid and economical.

Evaluation of Petrifilm Lactic Acid Bacteria Plates for Counting Lactic Acid Bacteria in Food.

PubMed

Kanagawa, Satomi; Ohshima, Chihiro; Takahashi, Hajime; Burenqiqige; Kikuchi, Misato; Sato, Fumina; Nakamura, Ayaka; Mohamed, Shimaa M; Kuda, Takashi; Kimura, Bon

2018-06-01

Although lactic acid bacteria (LAB) are used widely as starter cultures in the production of fermented foods, they are also responsible for food decay and deterioration. The undesirable growth of LAB in food causes spoilage, discoloration, and slime formation. Because of these adverse effects, food companies test for the presence of LAB in production areas and processed foods and consistently monitor the behavior of these bacteria. The 3M Petrifilm LAB Count Plates have recently been launched as a time-saving and simple-to-use plate designed for detecting and quantifying LAB. This study compares the abilities of Petrifilm LAB Count Plates and the de Man Rogosa Sharpe (MRS) agar medium to determine the LAB count in a variety of foods and swab samples collected from a food production area. Bacterial strains isolated from Petrifilm LAB Count Plates were identified by 16S rDNA sequence analysis to confirm the specificity of these plates for LAB. The results showed no significant difference in bacterial counts measured by using Petrifilm LAB Count Plates and MRS medium. Furthermore, all colonies growing on Petrifilm LAB Count Plates were confirmed to be LAB, while yeast colonies also formed in MRS medium. Petrifilm LAB Count Plates eliminated the plate preparation and plate inoculation steps, and the cultures could be started as soon as a diluted food sample was available. Food companies are required to establish quality controls and perform tests to check the quality of food products; the use of Petrifilm LAB Count Plates can simplify this testing process for food companies.

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

PubMed Central

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

2014-01-01

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

Development of a rapid optic bacteria detecting system based on ATP bioluminescence

NASA Astrophysics Data System (ADS)

Liu, Jun Tao; Luo, JinPing; Liu, XiaoHong; Cai, XinXia

2014-12-01

A rapid optic bacteria detecting system based on the principle of Adenosine triphosphate(ATP) bioluminescence was presented in this paper. This system consisted of bioluminescence-based biosensor and the high-sensitivity optic meter. A photon counting photomultiplier tube (PMT) module was used to improve the detection sensitivity, and a NIOS II/f processor based on a Field Programmable Gate Array(FPGA) was used to control the system. In this work, Micrococcus luteus were chosen as the test sample. Several Micrococcus luteus suspension with different concentration was tested by both T2011 and plate counting method. By comparing the two group results, an calibration curve was obtained from the bioluminescence intensity for Micrococcus luteus in the range of 2.3×102 ~ 2.3×106 CFU/mL with a good correlation coefficient of 0.960. An impacting Air microorganism sampler was used to capture Airborne Bacteria, and 8 samples were collected in different place. The TBC results of 8 samples by T2011 were between 10 ~ 2×103 cfu/mL, consistent with that of plate counting method, which indicated that 8 samples were between 10 ~ 3×103 cfu/mL. For total airborne bacteria count was small, correlation coefficient was poor. Also no significant difference was found between T2011 and plate counting method by statistical analyses.

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

PubMed

Stachelska, Milena A

2017-12-04

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

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

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

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

1994-05-01

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

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

PubMed

Fu, Xin; Huang, Kelong; Liu, Suqin

2010-02-01

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

A Hidden Pitfall in the Preparation of Agar Media Undermines Microorganism Cultivability

PubMed Central

Tanaka, Tomohiro; Kawasaki, Kosei; Daimon, Serina; Kitagawa, Wataru; Yamamoto, Kyosuke; Tamaki, Hideyuki; Tanaka, Michiko; Nakatsu, Cindy H.

2014-01-01

Microbiologists have been using agar growth medium for over 120 years. It revolutionized microbiology in the 1890s when microbiologists were seeking effective methods to isolate microorganisms, which led to the successful cultivation of microorganisms as single clones. But there has been a disparity between total cell counts and cultivable cell counts on plates, often referred to as the “great plate count anomaly,” that has long been a phenomenon that still remains unsolved. Here, we report that a common practice microbiologists have employed to prepare agar medium has a hidden pitfall: when phosphate was autoclaved together with agar to prepare solid growth media (PT medium), total colony counts were remarkably lower than those grown on agar plates in which phosphate and agar were separately autoclaved and mixed right before solidification (PS medium). We used a pure culture of Gemmatimonas aurantiaca T-27T and three representative sources of environmental samples, soil, sediment, and water, as inocula and compared colony counts between PT and PS agar plates. There were higher numbers of CFU on PS medium than on PT medium using G. aurantiaca or any of the environmental samples. Chemical analysis of PT agar plates suggested that hydrogen peroxide was contributing to growth inhibition. Comparison of 454 pyrosequences of the environmental samples to the isolates revealed that taxa grown on PS medium were more reflective of the original community structure than those grown on PT medium. Moreover, more hitherto-uncultivated microbes grew on PS than on PT medium. PMID:25281372

Evaluation of heterotrophic plate and chromogenic agar colony counting in water quality laboratories.

PubMed

Hallas, Gary; Monis, Paul

2015-01-01

The enumeration of bacteria using plate-based counts is a core technique used by food and water microbiology testing laboratories. However, manual counting of bacterial colonies is both time and labour intensive, can vary between operators and also requires manual entry of results into laboratory information management systems, which can be a source of data entry error. An alternative is to use automated digital colony counters, but there is a lack of peer-reviewed validation data to allow incorporation into standards. We compared the performance of digital counting technology (ProtoCOL3) against manual counting using criteria defined in internationally recognized standard methods. Digital colony counting provided a robust, standardized system suitable for adoption in a commercial testing environment. The digital technology has several advantages:•Improved measurement of uncertainty by using a standard and consistent counting methodology with less operator error.•Efficiency for labour and time (reduced cost).•Elimination of manual entry of data onto LIMS.•Faster result reporting to customers.

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

PubMed

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

2018-01-01

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

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

PubMed

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.

HETEROTROPHIC PLATE COUNT BACTERIA - WHAT IS THEIR SIGNIFICANCE IN DRINKING WATER?

EPA Science Inventory

The possible health significance of heterotrophic plate count (HPC) bacteria, also know in earlier terminology as standard plate count (SPC) bacteria, in drinking water has been debated for decades. While the literature documents the universal occurrence of HPC bacteria in soil, ...

A comparison of spatial analysis methods for the construction of topographic maps of retinal cell density.

PubMed

Garza-Gisholt, Eduardo; Hemmi, Jan M; Hart, Nathan S; Collin, Shaun P

2014-01-01

Topographic maps that illustrate variations in the density of different neuronal sub-types across the retina are valuable tools for understanding the adaptive significance of retinal specialisations in different species of vertebrates. To date, such maps have been created from raw count data that have been subjected to only limited analysis (linear interpolation) and, in many cases, have been presented as iso-density contour maps with contour lines that have been smoothed 'by eye'. With the use of stereological approach to count neuronal distribution, a more rigorous approach to analysing the count data is warranted and potentially provides a more accurate representation of the neuron distribution pattern. Moreover, a formal spatial analysis of retinal topography permits a more robust comparison of topographic maps within and between species. In this paper, we present a new R-script for analysing the topography of retinal neurons and compare methods of interpolating and smoothing count data for the construction of topographic maps. We compare four methods for spatial analysis of cell count data: Akima interpolation, thin plate spline interpolation, thin plate spline smoothing and Gaussian kernel smoothing. The use of interpolation 'respects' the observed data and simply calculates the intermediate values required to create iso-density contour maps. Interpolation preserves more of the data but, consequently includes outliers, sampling errors and/or other experimental artefacts. In contrast, smoothing the data reduces the 'noise' caused by artefacts and permits a clearer representation of the dominant, 'real' distribution. This is particularly useful where cell density gradients are shallow and small variations in local density may dramatically influence the perceived spatial pattern of neuronal topography. The thin plate spline and the Gaussian kernel methods both produce similar retinal topography maps but the smoothing parameters used may affect the outcome.

High spatial resolution detection of low-energy electrons using an event-counting method, application to point projection microscopy

NASA Astrophysics Data System (ADS)

Salançon, Evelyne; Degiovanni, Alain; Lapena, Laurent; Morin, Roger

2018-04-01

An event-counting method using a two-microchannel plate stack in a low-energy electron point projection microscope is implemented. 15 μm detector spatial resolution, i.e., the distance between first-neighbor microchannels, is demonstrated. This leads to a 7 times better microscope resolution. Compared to previous work with neutrons [Tremsin et al., Nucl. Instrum. Methods Phys. Res., Sect. A 592, 374 (2008)], the large number of detection events achieved with electrons shows that the local response of the detector is mainly governed by the angle between the hexagonal structures of the two microchannel plates. Using this method in point projection microscopy offers the prospect of working with a greater source-object distance (350 nm instead of 50 nm), advancing toward atomic resolution.

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

PubMed

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

2015-01-01

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

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

PubMed

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

2017-08-08

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

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

PubMed

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

2018-01-01

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

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

PubMed Central

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

2018-01-01

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

Comparison of the Petrifilm dry rehydratable film and conventional culture methods for enumeration of yeasts and molds in foods: collaborative study.

PubMed

Knight, M T; Newman, M C; Benzinger, M J; Neufang, K L; Agin, J R; McAllister, J S; Ramos, M

1997-01-01

A collaborative study was performed involving 18 laboratories and 6 food types to compare 3M Petrifilm yeast and mold count plates with the method described in the U.S. Food and Drug Administration's Bacteriological Analytical Manual. Four species of mold and 2 species of yeast were used to inoculate the following foods: hot dogs, corn meal, ketchup, orange juice, yogurt, and cake mix. Each collaborator received 15 samples of each food type: 5 low-level inoculations, 5 high-level inoculations, and 5 uninoculated samples. There was no significant difference between the means of the 2 methods for any product or inoculation level. The Petrifilm yeast and mold count plate method for enumeration of yeasts and molds in foods has been adopted first action by AOAC INTERNATIONAL.

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

PubMed

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

1972-04-01

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

Evaluation of the methods for enumerating coliform bacteria from water samples using precise reference standards.

PubMed

Wohlsen, T; Bates, J; Vesey, G; Robinson, W A; Katouli, M

2006-04-01

To use BioBall cultures as a precise reference standard to evaluate methods for enumeration of Escherichia coli and other coliform bacteria in water samples. Eight methods were evaluated including membrane filtration, standard plate count (pour and spread plate methods), defined substrate technology methods (Colilert and Colisure), the most probable number method and the Petrifilm disposable plate method. Escherichia coli and Enterobacter aerogenes BioBall cultures containing 30 organisms each were used. All tests were performed using 10 replicates. The mean recovery of both bacteria varied with the different methods employed. The best and most consistent results were obtained with Petrifilm and the pour plate method. Other methods either yielded a low recovery or showed significantly high variability between replicates. The BioBall is a very suitable quality control tool for evaluating the efficiency of methods for bacterial enumeration in water samples.

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

PubMed

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

2015-01-01

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

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

PubMed

Mosupye, F M; von Holy, A

2000-11-01

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

Absolute Enumeration of Probiotic Strains Lactobacillus acidophilus NCFM® and Bifidobacterium animalis subsp. lactis Bl-04 ® via Chip-Based Digital PCR.

PubMed

Hansen, Sarah J Z; Morovic, Wesley; DeMeules, Martha; Stahl, Buffy; Sindelar, Connie W

2018-01-01

The current standard for enumeration of probiotics to obtain colony forming units by plate counts has several drawbacks: long time to results, high variability and the inability to discern between bacterial strains. Accurate probiotic cell counts are important to confirm the delivery of a clinically documented dose for its associated health benefits. A method is described using chip-based digital PCR (cdPCR) to enumerate Bifidobacterium animalis subsp. lactis Bl-04 and Lactobacillus acidophilus NCFM both as single strains and in combination. Primers and probes were designed to differentiate the target strains against other strains of the same species using known single copy, genetic differences. The assay was optimized to include propidium monoazide pre-treatment to prevent amplification of DNA associated with dead probiotic cells as well as liberation of DNA from cells with intact membranes using bead beating. The resulting assay was able to successfully enumerate each strain whether alone or in multiplex. The cdPCR method had a 4 and 5% relative standard deviation (RSD) for Bl-04 and NCFM, respectively, making it more precise than plate counts with an industry accepted RSD of 15%. cdPCR has the potential to replace traditional plate counts because of its precision, strain specificity and the ability to obtain results in a matter of hours.

Experimental Study for Automatic Colony Counting System Based Onimage Processing

NASA Astrophysics Data System (ADS)

Fang, Junlong; Li, Wenzhe; Wang, Guoxin

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

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

PubMed Central

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

2015-01-01

A Japanese round-robin study revealed that analysts who used a dark-medium (DM) objective lens reported higher fiber counts from American Industrial Hygiene Association (AIHA) Proficiency Analytical Testing (PAT) chrysotile samples than those using a standard objective lens, but the cause of this difference was not investigated at that time. The purpose of this study is to determine any major source of this difference by performing two sets of round-robin studies. For the first round-robin study, 15 AIHA PAT samples (five each of chrysotile and amosite generated by water-suspended method, and five chrysotile generated by aerosolization method) were prepared with relocatable cover slips and examined by nine laboratories. A second round-robin study was then performed with six chrysotile field sample slides by six out of nine laboratories who participated in the first round-robin study. In addition, two phase-shift test slides to check analysts’ visibility and an eight-form diatom test plate to compare resolution between the two objectives were examined. For the AIHA PAT chrysotile reference slides, use of the DM objective resulted in consistently higher fiber counts (1.45 times for all data) than the standard objective (P-value < 0.05), regardless of the filter generation (water-suspension or aerosol) method. For the AIHA PAT amosite reference and chrysotile field sample slides, the fiber counts between the two objectives were not significantly different. No statistically significant differences were observed in the visibility of blocks of the test slides between the two objectives. Also, the DM and standard objectives showed no pattern of differences in viewing the fine lines and/or dots of each species images on the eight-form diatom test plate. Among various potential factors that might affect the analysts’ performance of fiber counts, this study supports the greater contrast caused by the different phase plate absorptions as the main cause of high counts for the AIHA PAT chrysotile slides using the DM objective. The comparison of fiber count ratios (DM/standard) between the AIHA PAT chrysotile samples and chrysotile field samples indicates that there is a fraction of fibers in the PAT samples approaching the theoretical limit of visibility of the phase-contrast microscope with 3-degree phase-shift. These fibers become more clearly visible through the greater contrast from the phase plate absorption of the DM objective. However, as such fibers are not present in field samples, no difference in counts between the two objectives was observed in this study. The DM objective, therefore, could be allowed for routine fiber counting as it will maintain continuity with risk assessments based on earlier phase-contrast microscopy fiber counts from field samples. Published standard methods would need to be modified to allow a higher aperture specification for the objective. PMID:25737333

Physiological responses of bacteria in biofilms to disinfection.

PubMed Central

Yu, F P; McFeters, G A

1994-01-01

In situ enumeration methods using fluorescent probes and a radioisotope labelling technique were applied to evaluate physiological changes of Klebsiella pneumoniae within biofilms after disinfection treatment. Chlorine (0.25 mg of free chlorine per liter [pH 7.2]) and monochloramine (1 mg/liter [pH 9.0]) were employed as disinfectants in the study. Two fluorgenic compounds, 5-cyano-2,3-ditolyl tetrazolium chloride and rhodamine 123, and tritiated uridine incorporation were chosen for assessment of physiological activities. Results obtained by these methods were compared with those from the plate count and direct viable count methods. 5-Cyano-2,3-ditolyl tetrazolium chloride is an indicator of bacterial respiratory activity, rhodamine 123 is incorporated into bacteria in response to transmembrane potential, and the incorporation of uridine represents the global RNA turnover rate. The results acquired by these methods following disinfection exposure showed a range of responses and suggested different physiological reactions in biofilms exposed to chlorine and monochloramine. The direct viable count response and respiratory activity were affected more by disinfection than were the transmembrane potential and RNA turnover rate on the basis of comparable efficiency as evaluated by plate count enumeration. Information revealed by these approaches can provide different physiological insights that may be used in evaluating the efficacy of biofilm disinfection. PMID:8074525

Population Screening Using Sewage Reveals Pan-Resistant Bacteria in Hospital and Community Samples.

PubMed

Meir-Gruber, Lital; Manor, Yossi; Gefen-Halevi, Shiraz; Hindiyeh, Musa Y; Mileguir, Fernando; Azar, Roberto; Smollan, Gill; Belausov, Natasha; Rahav, Galia; Shamiss, Ari; Mendelson, Ella; Keller, Nathan

2016-01-01

The presence of pan-resistant bacteria worldwide possesses a threat to global health. It is difficult to evaluate the extent of carriage of resistant bacteria in the population. Sewage sampling is a possible way to monitor populations. We evaluated the presence of pan-resistant bacteria in Israeli sewage collected from all over Israel, by modifying the pour plate method for heterotrophic plate count technique using commercial selective agar plates. This method enables convenient and fast sewage sampling and detection. We found that sewage in Israel contains multiple pan-resistant bacteria including carbapenemase resistant Enterobacteriacae carrying blaKPC and blaNDM-1, MRSA and VRE. blaKPC carrying Klebsiella pneumonia and Enterobacter cloacae were the most common Enterobacteriacae drug resistant bacteria found in the sewage locations we sampled. Klebsiella pneumonia, Enterobacter spp., Escherichia coli and Citrobacter spp. were the 4 main CRE isolated from Israeli sewage and also from clinical samples in our clinical microbiology laboratory. Hospitals and Community sewage had similar percentage of positive samplings for blaKPC and blaNDM-1. VRE was found to be more abundant in sewage in Israel than MRSA but there were more locations positive for MRSA and VRE bacteria in Hospital sewage than in the Community. Therefore, our upgrade of the pour plate method for heterotrophic plate count technique using commercial selective agar plates can be a useful tool for routine screening and monitoring of the population for pan-resistant bacteria using sewage.

Population Screening Using Sewage Reveals Pan-Resistant Bacteria in Hospital and Community Samples

PubMed Central

Mileguir, Fernando; Azar, Roberto; Smollan, Gill; Belausov, Natasha; Rahav, Galia; Shamiss, Ari; Mendelson, Ella; Keller, Nathan

2016-01-01

The presence of pan-resistant bacteria worldwide possesses a threat to global health. It is difficult to evaluate the extent of carriage of resistant bacteria in the population. Sewage sampling is a possible way to monitor populations. We evaluated the presence of pan-resistant bacteria in Israeli sewage collected from all over Israel, by modifying the pour plate method for heterotrophic plate count technique using commercial selective agar plates. This method enables convenient and fast sewage sampling and detection. We found that sewage in Israel contains multiple pan-resistant bacteria including carbapenemase resistant Enterobacteriacae carrying blaKPC and blaNDM-1, MRSA and VRE. blaKPC carrying Klebsiella pneumonia and Enterobacter cloacae were the most common Enterobacteriacae drug resistant bacteria found in the sewage locations we sampled. Klebsiella pneumonia, Enterobacter spp., Escherichia coli and Citrobacter spp. were the 4 main CRE isolated from Israeli sewage and also from clinical samples in our clinical microbiology laboratory. Hospitals and Community sewage had similar percentage of positive samplings for blaKPC and blaNDM-1. VRE was found to be more abundant in sewage in Israel than MRSA but there were more locations positive for MRSA and VRE bacteria in Hospital sewage than in the Community. Therefore, our upgrade of the pour plate method for heterotrophic plate count technique using commercial selective agar plates can be a useful tool for routine screening and monitoring of the population for pan-resistant bacteria using sewage. PMID:27780222

Comparison of viable plate count, turbidity measurement and real-time PCR for quantification of Porphyromonas gingivalis.

PubMed

Clais, S; Boulet, G; Van Kerckhoven, M; Lanckacker, E; Delputte, P; Maes, L; Cos, P

2015-01-01

The viable plate count (VPC) is considered as the reference method for bacterial enumeration in periodontal microbiology but shows some important limitations for anaerobic bacteria. As anaerobes such as Porphyromonas gingivalis are difficult to culture, VPC becomes time-consuming and less sensitive. Hence, efficient normalization of experimental data to bacterial cell count requires alternative rapid and reliable quantification methods. This study compared the performance of VPC with that of turbidity measurement and real-time PCR (qPCR) in an experimental context using highly concentrated bacterial suspensions. Our TaqMan-based qPCR assay for P. gingivalis 16S rRNA proved to be sensitive and specific. Turbidity measurements offer a fast method to assess P. gingivalis growth, but suffer from high variability and a limited dynamic range. VPC was very time-consuming and less repeatable than qPCR. Our study concludes that qPCR provides the most rapid and precise approach for P. gingivalis quantification. Although our data were gathered in a specific research context, we believe that our conclusions on the inferior performance of VPC and turbidity measurements in comparison to qPCR can be extended to other research and clinical settings and even to other difficult-to-culture micro-organisms. Various clinical and research settings require fast and reliable quantification of bacterial suspensions. The viable plate count method (VPC) is generally seen as 'the gold standard' for bacterial enumeration. However, VPC-based quantification of anaerobes such as Porphyromonas gingivalis is time-consuming due to their stringent growth requirements and shows poor repeatability. Comparison of VPC, turbidity measurement and TaqMan-based qPCR demonstrated that qPCR possesses important advantages regarding speed, accuracy and repeatability. © 2014 The Society for Applied Microbiology.

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

PubMed Central

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

1972-01-01

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

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

PubMed

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

2009-04-01

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

A Comparison of Spatial Analysis Methods for the Construction of Topographic Maps of Retinal Cell Density

PubMed Central

Garza-Gisholt, Eduardo; Hemmi, Jan M.; Hart, Nathan S.; Collin, Shaun P.

2014-01-01

Topographic maps that illustrate variations in the density of different neuronal sub-types across the retina are valuable tools for understanding the adaptive significance of retinal specialisations in different species of vertebrates. To date, such maps have been created from raw count data that have been subjected to only limited analysis (linear interpolation) and, in many cases, have been presented as iso-density contour maps with contour lines that have been smoothed ‘by eye’. With the use of stereological approach to count neuronal distribution, a more rigorous approach to analysing the count data is warranted and potentially provides a more accurate representation of the neuron distribution pattern. Moreover, a formal spatial analysis of retinal topography permits a more robust comparison of topographic maps within and between species. In this paper, we present a new R-script for analysing the topography of retinal neurons and compare methods of interpolating and smoothing count data for the construction of topographic maps. We compare four methods for spatial analysis of cell count data: Akima interpolation, thin plate spline interpolation, thin plate spline smoothing and Gaussian kernel smoothing. The use of interpolation ‘respects’ the observed data and simply calculates the intermediate values required to create iso-density contour maps. Interpolation preserves more of the data but, consequently includes outliers, sampling errors and/or other experimental artefacts. In contrast, smoothing the data reduces the ‘noise’ caused by artefacts and permits a clearer representation of the dominant, ‘real’ distribution. This is particularly useful where cell density gradients are shallow and small variations in local density may dramatically influence the perceived spatial pattern of neuronal topography. The thin plate spline and the Gaussian kernel methods both produce similar retinal topography maps but the smoothing parameters used may affect the outcome. PMID:24747568

Improved methods for the enumeration of heterotrophic bacteria in bottled mineral waters.

PubMed

Ramalho, R; Cunha, J; Teixeira, P; Gibbs, P A

2001-03-01

At this time the European Union regulations require that the heterotrophic plate counts (HPC) of mineral waters be assessed at two recovery temperatures: 22 degrees C for 72 h and 37 degrees C for 24 h. This procedure is time consuming and expensive. Development of new rapid methods for microbiological assessment of the microbial flora in the bottled water is an industry-driven need. The objectives of this work were to develop a method for the HPC that utilises only one recovery temperature and one incubation period and evaluate the use of, the LIVE/DEAD(R) BacLight Bacterial Viability Kit, 5-cyano-2,3-ditotyl tetrazolium chloride (CTC) and impedance methods to enumerate viable bacteria in bottled mineral water. Results showed that incubation at 30 degrees C could be used instead of incubation at 22 degrees C and 37 degrees C. Good correlation exists between counts at 30 degrees C and counts at 22 degrees C (r>0.90) and all the pathogens important in mineral water analyses grow similarly at 30 degrees C and 37 degrees C during 24 h. It was demonstrated that impedance methods might be useful to the mineral water industry as a rapid indicator of microbiological quality of the water. Results obtained with BacLight and CTC were similar to those obtained with plate counts.

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

NASA Astrophysics Data System (ADS)

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

2017-05-01

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

A miniaturized counting technique for anaerobic bacteria.

PubMed

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

1976-12-01

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

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

PubMed Central

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

2018-01-01

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

Optimization of Uranium Molecular Deposition for Alpha-Counting Sources

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

Monzo, Ellen; Parsons-Moss, Tashi; Genetti, Victoria

2016-12-12

Method development for molecular deposition of uranium onto aluminum 1100 plates was conducted with custom plating cells at Lawrence Livermore National Laboratory. The method development focused primarily on variation of electrode type, which was expected to directly influence plated sample homogeneity. Solid disc platinum and mesh platinum anodes were compared and data revealed that solid disc platinum anodes produced more homogenous uranium oxide films. However, the activity distribution also depended on the orientation of the platinum electrode relative to the aluminum cathode, starting current, and material composition of the plating cell. Experiments demonstrated these variables were difficult to control undermore » the conditions available. Variation of plating parameters among a series of ten deposited plates yielded variations up to 30% in deposition efficiency. Teflon particles were observed on samples plated in Teflon cells, which poses a problem for alpha activity measurements of the plates. Preliminary electropolishing and chemical polishing studies were also conducted on the aluminum 1100 cathode plates.« less

Selective cultivation and rapid detection of Staphylococcus aureus by computer vision.

PubMed

Wang, Yong; Yin, Yongguang; Zhang, Chaonan

2014-03-01

In this paper, we developed a selective growth medium and a more rapid detection method based on computer vision for selective isolation and identification of Staphylococcus aureus from foods. The selective medium consisted of tryptic soy broth basal medium, 3 inhibitors (NaCl, K2 TeO3 , and phenethyl alcohol), and 2 accelerators (sodium pyruvate and glycine). After 4 h of selective cultivation, bacterial detection was accomplished using computer vision. The total analysis time was 5 h. Compared to the Baird-Parker plate count method, which requires 4 to 5 d, this new detection method offers great time savings. Moreover, our novel method had a correlation coefficient of greater than 0.998 when compared with the Baird-Parker plate count method. The detection range for S. aureus was 10 to 10(7) CFU/mL. Our new, rapid detection method for microorganisms in foods has great potential for routine food safety control and microbiological detection applications. © 2014 Institute of Food Technologists®

Impedance technology reduces the enumeration time of Brettanomyces yeast during beer fermentation.

PubMed

van Wyk, Sanelle; Silva, Filipa V M

2016-12-01

Brettanomyces yeasts are increasingly being used to produce lambic style beers and craft beers with unique flavors. Currently, the industry monitors Brettanomyces bruxellensis using time consuming plate counting. B. bruxellensis is a fastidious slow growing organism, requiring five days of incubation at 30°C for visible growth on agar plates. Thus, a need exists to develop a quicker, feasible method to enumerate this yeast. The aim of this study was therefore to determine the feasibility of using the 'direct' and 'indirect' impedance methods for the enumeration of B. bruxellensis in beer and to monitor the growth of the yeast during fermentation. The impedance methods were able to decrease the incubation time of beer samples containing Brettanomyces from 120 h down to 2 and 84 h for samples containing 10 7 and 10 3 cfu/mL, respectively. The 'indirect' method was more successful than the 'direct' method, presenting a smaller error and wider detection range. Overall, the 'indirect' impedance method is a viable alternative to plate counting for the enumeration of yeasts in the brewing industry because it decreases preparation and incubation times, thereby increasing throughput and decreasing the chance of contamination. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

PubMed

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

2014-08-01

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

Comparison of methods for isolation and enumeration of thermophilic actinomycetes from dust.

PubMed Central

Treuhaft, M W; Arden Jones, M P

1982-01-01

Thermophilic actinomycetes are the primary sensitizing agents in farmer's lung disease. We compared dilution pour-plate and spread-plate methods for their usefulness in enumerating thermophilic actinomycetes in moldy silage dust and evaluated the ability of a nonquantitative gravity settling technique to recover thermophilic actinomycetes from moldy silage. Spread plates and pour plates yielded similar estimates of total thermophiles. Higher counts were observed on spread plates (P less than 0.05) for Thermoactinomyces candidus, Micropolyspora faeni, and Saccharomonospora viridis. M. faeni and S. viridis were less efficient than T. candidus in breaking through the agar of pour plates to form colonies which could be identified. Coefficients of variability were less than 10% for the two methods. The relative proportion of organisms recovered by the settling method correlated well with that recovered on spread plates for M. faeni (r = 0.79), S. viridis (r = 0.88), and Thermomonospora spp. (r = 0.79), but not well for T. candidus (r = 0.28). When sophisticated air-sampling equipment is not available, dilution spread plates of dust washings provide a reproducible method for enumerating a broad range of thermophilic actinomycetes of interest. The gravity settling method is a simple, rapid alternative when isolation is all that is required. PMID:6761363

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

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

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

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

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

USDA-ARS?s Scientific Manuscript database

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

A high-throughput AO/PI-based cell concentration and viability detection method using the Celigo image cytometry.

PubMed

Chan, Leo Li-Ying; Smith, Tim; Kumph, Kendra A; Kuksin, Dmitry; Kessel, Sarah; Déry, Olivier; Cribbes, Scott; Lai, Ning; Qiu, Jean

2016-10-01

To ensure cell-based assays are performed properly, both cell concentration and viability have to be determined so that the data can be normalized to generate meaningful and comparable results. Cell-based assays performed in immuno-oncology, toxicology, or bioprocessing research often require measuring of multiple samples and conditions, thus the current automated cell counter that uses single disposable counting slides is not practical for high-throughput screening assays. In the recent years, a plate-based image cytometry system has been developed for high-throughput biomolecular screening assays. In this work, we demonstrate a high-throughput AO/PI-based cell concentration and viability method using the Celigo image cytometer. First, we validate the method by comparing directly to Cellometer automated cell counter. Next, cell concentration dynamic range, viability dynamic range, and consistency are determined. The high-throughput AO/PI method described here allows for 96-well to 384-well plate samples to be analyzed in less than 7 min, which greatly reduces the time required for the single sample-based automated cell counter. In addition, this method can improve the efficiency for high-throughput screening assays, where multiple cell counts and viability measurements are needed prior to performing assays such as flow cytometry, ELISA, or simply plating cells for cell culture.

Quick counting method for estimating the number of viable microbes on food and food processing equipment.

PubMed

Winter, F H; York, G K; el-Nakhal, H

1971-07-01

A rapid method for estimating the extent of microbial contamination on food and on food processing equipment is described. Microbial cells are rinsed from food or swab samples with sterile diluent and concentrated on the surface of membrane filters. The filters are incubated on a suitable bacteriological medium for 4 hr at 30 C, heated at 105 C for 5 min, and stained. The membranes are then dried at 60 C for 15 min, rendered transparent with immersion oil, and examined microscopically. Data obtained by the rapid method were compared with counts of the same samples determined by the standard plate count method. Over 60 comparisons resulted in a correlation coefficient of 0.906. Because the rapid technique can provide reliable microbiological count information in extremely short times, it can be a most useful tool in the routine evaluation of microbial contamination of food processing facilities and for some foods.

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

PubMed

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

2007-04-01

Attachment of Shiga toxigenic Escherichia coli (STEC) to surfaces and the formation of biofilms may enhance persistence in a food processing environment and present a risk of contaminating products. Seven strains of STEC and three non-STEC strains were selected to compare two biofilm quantification methods; epifluorescence microscopy on stainless steel (SS) and a microtitre plate assay. The influence of prior growth in planktonic (nutrient broth) and sessile (nutrient agar) culture on biofilm production, as well as expression of surface structures and the possession of antigen 43 (encoded by agn43) on biofilm formation were also investigated. Biofilms were produced in diluted nutrient broth at 25 degrees C for 24 and 48 h. Curli expression was determined using congo red indicator agar, while the presence of agn43 was determined using polymerase chain reaction. No correlation was found between counts for epifluorescence microscopy on SS and the absorbance values obtained with the microtitre plate method for planktonic and sessile grown cultures. Different abilities of individual STEC strains to attach to SS and microtitre plates were found with some strains attaching better to each surface following growth in either planktonic or sessile culture. All O157 STEC strains had low biofilm counts on SS for planktonic and sessile grown cultures; however, one STEC O157:H- strain (EC516) had significantly greater (p<0.05) biofilm production on microtitre plates compared to the other O157 STEC strains. EC516 and other STEC (O174:H21 and O91:H21) strains expressing curli fimbriae were found to produce significantly greater (p<0.05) biofilms on microtitre plates compared to the non-curli expressing strains. No relationship was found between the production of type-I fimbriae, motility, agn43 and bacterial physicochemical properties (previously determined) and biofilm formation on SS or microtitre plates. Variations between the two biofilm determination methods may suggest that the biofilm production on microtitre plates may not be appropriate to represent other surfaces such as SS and that caution should be taken when selecting a method to quantify biofilm production on a surface.

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

NASA Astrophysics Data System (ADS)

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

2017-03-01

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

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

PubMed

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

2013-08-01

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

Rapid High-Throughput Assessment of Aerobic Bacteria in Complex Samples by Fluorescence-Based Oxygen Respirometry

PubMed Central

O'Mahony, Fiach C.; Papkovsky, Dmitri B.

2006-01-01

A simple method has been developed for the analysis of aerobic bacteria in complex samples such as broth and food homogenates. It employs commercial phosphorescent oxygen-sensitive probes to monitor oxygen consumption of samples containing bacteria using standard microtiter plates and fluorescence plate readers. As bacteria grow in aqueous medium, at certain points they begin to deplete dissolved oxygen, which is seen as an increase in probe fluorescence above baseline signal. The time required to reach threshold signal is used to either enumerate bacteria based on a predetermined calibration or to assess the effects of various effectors on the growth of test bacteria by comparison with an untreated control. This method allows for the sensitive (down to a single cell), rapid (0.5 to 12 h) enumeration of aerobic bacteria without the need to conduct lengthy (48 to 72 h) and tedious colony counts on agar plates. It also allows for screening a wide range of chemical and environmental samples for their toxicity. These assays have been validated with different bacteria, including Escherichia coli, Micrococcus luteus, and Pseudomonas fluorescens, with the enumeration of total viable counts in broth and industrial food samples (packaged ham, chicken, and mince meat), and comparison with established agar plating and optical-density-at-600-nm assays has been given. PMID:16461677

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

PubMed

Garcés-Vega, Francisco; Marks, Bradley P

2014-08-01

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

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

NASA Astrophysics Data System (ADS)

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

2018-01-01

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

Comparison of solid-phase cytometry and the plate count method for the evaluation of the survival of bacteria in pharmaceutical oils.

PubMed

De Prijck, K; Peeters, E; Nelis, H J

2008-12-01

To compare the survival of four bacterial strains (Escherichia coli, Proteus mirabilis, Staphylococcus aureus, Pseudomonas aeruginosa) in pharmaceutical oils, including jojoba oil/tea tree oil, carbol oil, jojoba oil and sesame oil. Oils were spiked with the test bacteria in a concentration of 10(4) CFU ml(-1). Bacteria were extracted from oils with phosphate-buffered saline containing 0.5% Tween 20. Aliquots of the pooled water layers were analysed by solid-phase cytometry and plate counting. Plate counts dropped to zero for all test strains exposed for 24 h to three of the four oils. In contrast, significant numbers of viable cells were still detected by SPC, except in the jojoba oil/tea tree oil mixture and partly in sesame oil. Exposure of bacteria for 24 h to the two oils containing an antimicrobial led to a loss of their culturability but not necessarily of their viability. The antibacterial activity of the jojoba oil/tea tree oil mixture supersedes that of carbol oil. These in vitro data suggest that the jojoba oil/tea tree oil mixture more than carbol oil inhibits bacterial proliferation when used for intermittent self-catherization.

Death of the Escherichia coli K-12 strain W3110 in soil and water.

PubMed Central

Bogosian, G; Sammons, L E; Morris, P J; O'Neil, J P; Heitkamp, M A; Weber, D B

1996-01-01

Whether Escherichia coli K-12 strain W3110 can enter the "viable but nonculturable" state was studied with sterile and nonsterile water and soil at various temperatures. In nonsterile river water, the plate counts of added E. coli cells dropped to less than 10 CFU/ml in less than 10 days. Acridine orange direct counts, direct viable counts, most-probable-number estimates, and PCR analyses indicated that the added E. coli cells were disappearing from the water in parallel with the number of CFU. Similar results were obtained with nonsterile soil, although the decline of the added E. coli was slower. In sterile water or soil, the added E. coli persisted for much longer, often without any decline in the plate counts even after 50 days. In sterile river water at 37 degrees C and sterile artificial seawater at 20 and 37 degrees C, the plate counts declined by 3 to 5 orders of magnitude, while the acridine orange direct counts remained unchanged. However, direct viable counts and various resuscitation studies all indicated that the nonculturable cells were nonviable. Thus, in either sterile or nonsterile water and soil, the decline in plate counts of E. coli K-12 strain W3110 is not due to the cells entering the viable but nonculturable state, but is simply due to their death. PMID:8900002

Protocol for evaluating the efficacy of cetylpyridinium chloride as a beef hide intervention.

PubMed

Bosilevac, Joseph M; Wheeler, Tommy L; Rivera-Betancourt, Mildred; Nou, Xiangwu; Arthur, Terrance M; Shackelford, Steven D; Kent, Matthew P; Jaroni, Divya; Osborn, Matthew S; Rossman, Michelle; Reagan, James O; Koohmaraie, Mohammad

2004-02-01

The objective of this study was to establish the necessary protocols and assess the efficacy of cetylpyridinium chloride (CPC) as an antimicrobial intervention on beef cattle hides. Experiments using CPC were conducted to determine (i) the methods of neutralization needed to obtain valid efficacy measurements, (ii) the effect of concentration and dwell time after treatment, (iii) the effect of CPC on hide and carcass microbial populations when cattle were treated at a feedlot and then transported to a processing facility for harvest, and (iv) the effectiveness of spray pressure and two-spray combinations of CPC and water to reduce hide microbial populations. Residual CPC in hide sponge samples prevented bacterial growth. Dey-Engley neutralization media at 7.8% and a centrifugation step were necessary to overcome this problem. All dwell times, ranging from 30 s to 4 h, after 1% CPC application to cattle hides resulted in aerobic plate counts and Enterobacteriaceae counts 1.5 log CFU/100 cm2 lower than controls. The most effective dose of CPC was 1%, which reduced aerobic plate counts and Enterobacteriaceae counts 2 and 1 log CFU/100 cm2, respectively. Low-pressure application of 1% CPC at the feedlot, transport to the processing facility, and harvest within 5 h of application resulted in no effect on Escherichia coli O157 prevalence on hides or preevisceration carcasses. Two high-pressure CPC washes lowered aerobic plate counts and Enterobacteriaceae counts by 4 log CFU/100 cm2, and two medium-pressure CPC washes were only slightly less effective. These results indicate that under the proper conditions, CPC may still be effective for reducing microbial populations on cattle hides. Further study is warranted to determine if this effect will result in reduction of hide-to-carcass contamination during processing.

Microbiological and molecular characterization of commercially available probiotics containing Bacillus clausii from India and Pakistan.

PubMed

Patrone, Vania; Molinari, Paola; Morelli, Lorenzo

2016-11-21

Probiotics are actively used for treatment of diarrhoea, respiratory infections, and prevention of infectious gastrointestinal diseases. The efficacy of probiotics is due to strain-specific features and the number of viable cells; however, several reports of deviations from the label in the actual content of strains in probiotic products are a matter of concern. Most of the available data on quality focuses on probiotic products containing lactobacilli and/or bifidobacteria, while very few data are available on spore-forming probiotics. The present study evaluates the label claims for spore count and species identification in five commercial probiotic products marketed in India and Pakistan that claim to contain Bacillus clausii: Tufpro, Ecogro, Enterogermina, Entromax, and Ospor. Bacterial enumeration from three batches was done by microbiological plating methods by two independent operators. Species identification was done using PCR amplification and sequence analysis of the 16S rRNA gene, and determination of the total amount of species present in the products was done using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analysis followed by DNA sequencing of the excised bands. Plate count methods demonstrated poor correlations between quantitative label indications and bacteria recovered from plates for Tufpro, Ecogro, and Ospor. The 16S rRNA analysis performed on bacteria isolated from plate counts showed that only Enterogermina and Ospor contained homogenous B. clausii. PCR-DGGE analysis revealed that only Enterogermina had a homogenous B. clausii population while other products had mixed bacterial populations. In conclusion, the current analysis clearly demonstrates that of the five analysed commercial probiotics, only Enterogermina followed the label claims. Copyright © 2016 Elsevier B.V. All rights reserved.

AUTOMATIC COUNTER

DOEpatents

Robinson, H.P.

1960-06-01

An automatic counter of alpha particle tracks recorded by a sensitive emulsion of a photographic plate is described. The counter includes a source of mcdulated dark-field illumination for developing light flashes from the recorded particle tracks as the photographic plate is automatically scanned in narrow strips. Photoelectric means convert the light flashes to proportional current pulses for application to an electronic counting circuit. Photoelectric means are further provided for developing a phase reference signal from the photographic plate in such a manner that signals arising from particle tracks not parallel to the edge of the plate are out of phase with the reference signal. The counting circuit includes provision for rejecting the out-of-phase signals resulting from unoriented tracks as well as signals resulting from spurious marks on the plate such as scratches, dust or grain clumpings, etc. The output of the circuit is hence indicative only of the tracks that would be counted by a human operator.

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

PubMed

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 < 0.05) decrease in 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 < 0.05) reduction in the bacteriological cell count. This analysis evidenced a significant time effect in the average log bacteriological cell count. Although 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.

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

PubMed

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

2010-01-01

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

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

PubMed

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

2010-02-01

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

Rapid enumeration of low numbers of moulds in tea based drinks using an automated system.

PubMed

Tanaka, Kouichi; Yamaguchi, Nobuyasu; Baba, Takashi; Amano, Norihide; Nasu, Masao

2011-01-31

Aseptically prepared cold drinks based on tea have become popular worldwide. Contamination of these drinks with harmful microbes is a potential health problem because such drinks are kept free from preservatives to maximize aroma and flavour. Heat-tolerant conidia and ascospores of fungi can survive pasteurization, and need to be detected as quickly as possible. We were able to rapidly and accurately detect low numbers of conidia and ascospores in tea-based drinks using fluorescent staining followed by an automated counting system. Conidia or ascospores were inoculated into green tea and oolong tea, and samples were immediately filtered through nitrocellulose membranes (pore size: 0.8 μm) to concentrate fungal propagules. These were transferred onto potato dextrose agar and incubated for 23 h at 28 °C. Fungi germinating on the membranes were fluorescently stained for 30 min. The stained mycelia were counted selectively within 90s using an automated counting system (MGS-10LD; Chuo Electric Works, Osaka, Japan). Very low numbers (1 CFU/100ml) of conidia or ascospores could be rapidly counted, in contrast to traditional labour intensive techniques. All tested mould strains were detected within 24h while conventional plate counting required 72 h for colony enumeration. Counts of slow-growing fungi (Cladosporium cladosporioides) obtained by automated counting and by conventional plate counting were close (r(2) = 0.986). Our combination of methods enables counting of both fast- and slow-growing fungi, and should be useful for microbiological quality control of tea-based and also other drinks. Copyright © 2011 Elsevier B.V. All rights reserved.

Quality control of fifteen probiotic products containing Saccharomyces boulardii.

PubMed

Vanhee, L M E; Goemé, F; Nelis, H J; Coenye, T

2010-11-01

The yeast Saccharomyces boulardii is used as a probiotic for the prevention and treatment of diarrhoea. In this study, the quality of 15 probiotic products containing S. boulardii was verified. Using microsatellite typing, the identity of all Saccharomyces strains in the products was confirmed as S. boulardii. Additionally, solid-phase cytometry (SPC) and a plate method were used to enumerate S. boulardii cells. SPC was not only able to produce results more rapidly than plating (4h compared to 48h) but the cell counts obtained with SPC were significantly higher than the plate counts. Finally, we found that <1% of the S. boulardii cells survived 120min in gastric conditions and storage for 3months at 40°C with 75% relative humidity. We developed a SPC method for the quantification of viable S. boulardii cells in probiotics. Additionally, we demonstrated that gastric conditions and storage have a marked effect on the viability of the yeast cells.   To our knowledge, this is the first time SPC is used for the quality control of probiotics with S. boulardii. Additionally, we demonstrated the need for gastric protection and accurate storage. © 2010 The Authors. Journal of Applied Microbiology © 2010 The Society for Applied Microbiology.

Regulation of Glucose Utilization by Estradiol in Breast Cancer

DTIC Science & Technology

2014-10-01

counts/min) weremeasured using 350 l of lysate. Protein concentration was determined using the BCA assay according to the manufacturer’s...instructions and measured on a Powerwave XS plate reader (Biotek). Counts were normalized to protein concentration. Glycolysis Assay—MCF-7 cells growing in 6...determined using the BCA assay according to the manufacturer’s instructions and mea- sured on a Powerwave XS plate reader. Counts were normal- ized to

Detection of coliform bacteria and Escherichia coli by multiplex polymerase chain reaction: comparison with defined substrate and plating methods for water quality monitoring.

PubMed Central

Bej, A K; McCarty, S C; Atlas, R M

1991-01-01

Multiplex polymerase chain reaction (PCR) and gene probe detection of target lacZ and uidA genes were used to detect total coliform bacteria and Escherichia coli, respectively, for determining water quality. In tests of environmental water samples, the lacZ PCR method gave results statistically equivalent to those of the plate count and defined substrate methods accepted by the U.S. Environmental Protection Agency for water quality monitoring and the uidA PCR method was more sensitive than 4-methylumbelliferyl-beta-D-glucuronide-based defined substrate tests for specific detection of E. coli. Images PMID:1768116

Evaluation of potassium-clavulanate-supplemented modified charcoal-cefoperazone-deoxycholate agar for enumeration of Campylobacter in chicken carcass rinse.

PubMed

Chon, Jung-Whan; Kim, Hong-Seok; Kim, Hyunsook; Oh, Deog-Hwan; Seo, Kun-Ho

2014-05-01

Potassium-clavulanate-supplemented modified charcoal-cefoperazone-deoxycholate agar (C-mCCDA) that was described in our previous study was compared with original mCCDA for the enumeration of Campylobacter in pure culture and chicken carcass rinse. The quantitative detection of viable Campylobacter cells from a pure culture, plated on C-mCCDA, is statistically similar (P > 0.05) to mCCDA. In total, 120 chickens were rinsed using 400 mL buffered peptone water. The rinses were inoculated onto C-mCCDA and mCCDA followed by incubation at 42 °C for 48 h. There was no statistical difference between C-mCCDA (45 of 120 plates; mean count, 145.5 CFU/mL) and normal mCCDA (46 of 120 plates; mean count, 160.8 CFU/mL) in the isolation rate and recovery of Campylobacter (P > 0.05) from chicken carcass rinse. The Pearson correlation coefficient value for the number of Campylobacter cells recovered in the 2 media was 0.942. However, the selectivity was much better on C-mCCDA than on mCCDA plates (P < 0.05). Significantly fewer C-mCCDA plates (33 out of 120 plates; mean count, 1.9 CFU/mL) were contaminated with non-Campylobacter cells than the normal mCCDA plates (67 out of 120 plates; mean count, 27.1 CFU/mL). The C-mCCDA may provide improved results for enumeration of Campylobacter in chicken meat alternative to mCCDA with its increased selectivity the modified agar possess. © 2014 Institute of Food Technologists®

Evaluation of antimicrobial activity of selected plant extracts by rapid XTT colorimetry and bacterial enumeration.

PubMed

Al-Bakri, Amal G; Afifi, Fatma U

2007-01-01

The aim of this study was to screen and evaluate the antimicrobial activity of indigenous Jordanian plant extracts, dissolved in dimethylsulfoxide, using the rapid XTT assay and viable count methods. XTT rapid assay was used for the initial screening of antimicrobial activity for the plant extracts. Antimicrobial activity of potentially active plant extracts was further assessed using the "viable plate count" method. Four degrees of antimicrobial activity (high, moderate, weak and inactive) against Bacillus subtilis, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa, respectively, were recorded. The plant extracts of Hypericum triquetrifolium, Ballota undulata, Ruta chalepensis, Ononis natrix, Paronychia argentea and Marrubium vulgare had shown promising antimicrobial activity. This study showed that while both XTT and viable count methods are comparable when estimating the overall antimicrobial activity of experimental substances, there is no strong linear correlation between the two methods.

Direct quantification and distribution of tetracycline-resistant genes in meat samples by real-time polymerase chain reaction.

PubMed

Guarddon, Mónica; Miranda, Jose M; Vázquez, Beatriz I; Cepeda, Alberto; Franco, Carlos M

2012-07-01

The evolution of antimicrobial-resistant bacteria has become a threat to food safety and methods to control them are necessary. Counts of tetracycline-resistant (TR) bacteria by microbiological methods were compared with those obtained by quantitative PCR (qPCR) in 80 meat samples. TR Enterobacteriaceae counts were similar between the count plate method and qPCR (P= 0.24), whereas TR aerobic mesophilic bacteria counts were significantly higher by the microbiological method (P < 0.001). The distribution of tetA and tetB genes was investigated in different types of meat. tetA was detected in chicken meat (40%), turkey meat (100%), pork (20%), and beef (40%) samples, whereas tetB was detected in chicken meat (45%), turkey meat (70%), pork (30%), and beef (35%) samples. The presence of tetracycline residues was also investigated by a receptor assay. This study offers an alternative and rapid method for monitoring the presence of TR bacteria in meat and furthers the understanding of the distribution of tetA and tetB genes. © 2012 Institute of Food Technologists®

Enumeration procedure for monitoring test microbe populations on inoculated carriers in AOAC use-dilution methods.

PubMed

Tomasino, Stephen F; Fiumara, Rebecca M; Cottrill, Michele P

2006-01-01

The AOAC Use-Dilution methods do not provide procedures to enumerate the test microbe on stainless steel carriers (penicylinders) or guidance on the expected target populations of the test microbe (i.e., a performance standard). This report describes the procedures used by the U.S. Environmental Protection Agency to enumerate the test microbe (carrier counts) associated with conducting the Use-Dilution method with Staphylococcus aureus (Method 955.15) and Pseudomonas aeruginosa (Method 964.02) and the examination of historical data. The carrier count procedure involves the random selection of carriers, shearing bacterial cells from the carrier surface through sonication, and plating of serially diluted inoculum on trypticase soy agar. For each Use-Dilution test conducted, the official AOAC method was strictly followed for carrier preparation, culture initiation, test culture preparation, and carrier inoculation steps. Carrier count data from 78 Use-Dilution tests conducted over a 6-year period were compiled and analyzed. A mean carrier count of 6.6 logs (approximately 4.0 x 10(6) colony-forming units/carrier) was calculated for both S. aureus and P. aeruginosa. Of the mean values, 95% fell within +/- 2 repeatability standard deviations. The enumeration procedure and target carrier counts are desirable for standardizing the Use-Dilution methods, increasing their reproducibility, and ensuring the quality of the data.

Blood cell counting and classification by nonflowing laser light scattering method

NASA Astrophysics Data System (ADS)

Yang, Ye; Zhang, Zhenxi; Yang, Xinhui; Jiang, Dazong; Yeo, Joon Hock

1999-11-01

A new non-flowing laser light scattering method for counting and classifying blood cells is presented. A linear charge- coupled device with 1024 elements is used to detect the scattered light intensity distribution of the blood cells. A pinhole plate is combined with the CCD to compete the focusing of the measurement system. An isotropic sphere is used to simulate the blood cell. Mie theory is used to describe the scattering of blood cells. In order to inverse the size distribution of blood cells from their scattered light intensity distribution, Powell method combined with precision punishment method is used as a dependent model method for measurement red blood cells and blood plates. Non-negative constraint least square method combined with Powell method and precision punishment method is used as an independent model for measuring white blood cells. The size distributions of white blood cells and red blood cells, and the mean diameter of red blood cells are measured by this method. White blood cells can be divided into three classes: lymphocytes, middle-sized cells and neutrocytes according to their sizes. And the number of blood cells in unit volume can also be measured by the linear dependence of blood cells concentration on scattered light intensity.

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

Andre, M.; Hugot, G.

A procedure is described for the detection of alpha -emitting contaminants of the respiratory tract. The method is useful when contamination from inhaled Pu/sup 239/ is suspected. Sputum, collected early in the morning, is dissolved in mineral acids and then evaporated on a stainiess steel plate. Radioautograms are prepared by placing the plate in contact with a nuclear emulsion. The number of tracks appearing after 24 and 48 hrs and 7 days is counted. The results are useful in estimating internal contamination and in following removal from the respiratory tract. (C.H.)

Advantageous Direct Quantification of Viable Closely Related Probiotics in Petit-Suisse Cheeses under In Vitro Gastrointestinal Conditions by Propidium Monoazide - qPCR

PubMed Central

Villarreal, Martha Lissete Morales; Padilha, Marina; Vieira, Antonio Diogo Silva; Franco, Bernadette Dora Gombossy de Melo; Martinez, Rafael Chacon Ruiz; Saad, Susana Marta Isay

2013-01-01

Species-specific Quantitative Real Time PCR (qPCR) alone and combined with the use of propidium monoazide (PMA) were used along with the plate count method to evaluate the survival of the probiotic strains Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis Bb-12, and the bacteriocinogenic and potentially probiotic strain Lactobacillus sakei subsp. sakei 2a in synbiotic (F1) and probiotic (F2) petit-suisse cheeses exposed throughout shelf-life to in vitro simulated gastrointestinal tract conditions. The three strains studied showed a reduction in their viability after the 6 h assay. Bb-12 displayed the highest survival capacity, above 72.6 and 74.6% of the initial populations, respectively, by plate count and PMA-qPCR, maintaining population levels in the range or above 6 log CFU/g. The prebiotic mix of inulin and FOS did not offer any additional protection for the strains against the simulated gastrointestinal environment. The microorganisms' populations were comparable among the three methods at the initial time of the assay, confirming the presence of mainly viable and culturable cells. However, with the intensification of the stress induced throughout the various stages of the in vitro test, the differences among the methods increased. The qPCR was not a reliable enumeration method for the quantification of intact bacterial populations, mixed with large numbers of injured and dead bacteria, as confirmed by the scanning electron microscopy results. Furthermore, bacteria plate counts were much lower (P<0.05) than with the PMA-qPCR method, suggesting the accumulation of stressed or dead microorganisms unable to form colonies. The use of PMA overcame the qPCR inability to differentiate between dead and alive cells. The combination of PMA and species-specific qPCR in this study allowed a quick and unequivocal way of enumeration of viable closely related species incorporated into probiotic and synbiotic petit-suisse cheeses and under stress conditions. PMID:24358142

Flocked nylon swabs versus RODAC plates for detection of multidrug-resistant organisms on environmental surfaces in intensive care units.

PubMed

Okamoto, K; Rhee, Y; Schoeny, M; Lolans, K; Cheng, J; Reddy, S; Weinstein, R A; Hayden, M K; Popovich, K J

2018-01-01

To compare two culture methods [nylon fiber flocked swabs with broth enrichment versus RODAC ('replicate organism detection and counting') plates] for recovery of multidrug-resistant organisms, 780 environmental surfaces in 63 rooms of patients on contact precautions in four intensive care units at one hospital were examined. Among sites that had at least one positive culture, swab culture with broth enrichment detected the target organisms more frequently than RODAC plates (37.5% vs 26.0%, P = 0.06). There was moderate agreement between the two methods (κ = 0.44) with agreement better for small or flat surfaces compared to large or irregular surfaces. Copyright © 2017 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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

1997-08-01

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

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

PubMed

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

2016-11-01

New technologies are emerging that may help individuals engage in healthier eating behaviors. One paradigm to test the efficacy of a technology is to determine its effect relative to environment cues that are known to cause individuals to overeat. The purpose of this work was to independently investigate two questions: How does the presence of a technology that provides bite count feedback alter eating behavior? and, How does the presence of a technology that provides bite count feedback paired with a goal alter eating behavior? Two studies investigated these research questions. The first study tested the effects of a large and small plate crossed with the presence or absence of a device that provided bite count feedback on intake. The second study tested the effects of a bite count goal with bite count feedback, again crossed with plate size, on intake. Both studies used a 2×2 between-subjects design. In the first study, 94 subjects (62 women aged 19.0±1.6 years with body mass index [BMI] 23.04±3.6) consumed lunch in a laboratory. The second study examined 99 subjects (56 women aged 18.5±1.5 years with BMI 22.73±2.70) under the same conditions. In both studies subjects consumed a single-course meal, using either a small or large plate. In the first study participants either wore or did not wear an automated bite counting device. In the second study all participants wore the bite counting device and were given either a low bite count goal (12 bites) or a high bite count goal (22 bites). Effect of plate size, feedback, and goal on consumption (grams) and number of bites taken were assessed using 2×2 analyses of variance. As adjunct measures, the effects of serving size, bite size (grams per bite), postmeal satiety, and satiety change were also assessed. In the first study there was a main effect of plate size on grams consumed and number of bites taken such that eating from a large plate led to greater consumption (P=0.001) and a greater number of bites (P=0.001). There was also a main effect of feedback on consumption and number of bites taken such that those who received feedback consumed less (P=0.011) and took fewer bites (P<0.001). In the second study there was a main effect of plate size on consumption such that those eating from a large plate consumed more (P=0.003) but did not take more bites. Further analysis revealed a main effect of goal on number of bites taken such that those who received the low goal took fewer bites (P<0.001) but did not consume less. Providing feedback on the number of bites taken from a wearable intake monitor can reduce overall intake during a single meal. Regarding the first research question, providing feedback significantly reduced intake in both plate size groups and reduced the overall number of bites taken. Regarding the second research question, participants were successful in eating to their goals. However, individuals in the low goal condition appeared to compensate for the restricted goal by taking larger bites, leading to comparable levels of consumption between the low and high goal groups. Hence, the interaction of technology with goals should be considered when introducing a health intervention. Copyright © 2016 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

[Endotoxin Contamination and Correlation with Other Water Quality Parameters of Groundwater from Self-Contained Wells in Beijing].

PubMed

Zhang, Can; Liu, Wen-jun; Ao, Lu; Shi, Yun; An, Dai-zhi; Liu, Zhi-ping

2015-12-01

A survey of endotoxin activity in groundwater from 14 self-contained wells in PLA units stationed in Beijing was conducted by the kinetic-turbid assay of Tachypleus Amebocyte Lysate (TAL). Bacteriological parameters, including total cell counts detected by flow cytometry, heterotrophic plate counts (HPC), standard plate counts and total coliforms were analyzed. Additionally, suspended particles, turbidity, dissolved organic carbon (DOC), and UV₂₅₄ were investigated. Total endotoxin activities ranged from 0. 15 to 13.20 EU · mL⁻¹, free endotoxin activities ranged from 0.10 to 5.29 EU · mL⁻¹ and bound endotoxin activities ranged from 0.01 to 8.60 EU · mL⁻¹. Most of the endotoxins in heavily contaminated groundwater existed as bound endotoxins. As for total endotoxins, the sequence of correlation coefficients with other parameters was total cell counts (r = 0.88 ) > HPC (r = 0.79) > DOC (r = 0.77) > UV₂₅₄ (r = 0.57) > total coliforms (r = 0.50) > standard plate counts (r = 0.49) = turbidity (r = 0. 49) > total particles (r = 0.41). The sequence of correlations of the bound endotoxins with other parameters was total cell counts (r = 0.81) > HPC (r = 0.66) > total coliforms (r = 0.65) > turbidity (r = 0.62) > total particles (r = 0.58) > standard plate counts (r = 0.22). Free endotoxins were correlated with DOC and UV₂₅₄, r = 0.58 and 0.26, respectively. Result showed free endotoxins had a higher correlation with DOC, and a lower correlation with UV₂₅₄.

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

PubMed

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

2011-07-01

Legionella species are the causative agents of human legionellosis, and bathing facilities have been identified as the sources of infection in several outbreaks in Japan. Researchers in Japan have recently reported evidence of significant associations between bacterial counts and the occurrence of Legionella in bathing facilities and in a hot tub model. A convenient and quantitative bacterial enumeration method is therefore required as an indicator of Legionella contamination or disinfection to replace existing methods such as time-consuming Legionella culture and expensive Legionella-DNA amplification. In this study, we developed a rapid detection method (RDM) to monitor the risk of Legionella using an automated microbial analyzing device based on flow cytometry techniques to measure the total number of bacteria in water samples within two minutes, by detecting typical patterns of scattered light and fluorescence. We first compared the results of our RDM with plate counting results for five filtered hot spring water samples spiked with three species of bacteria, including Legionella. Inactivation of these samples by chlorine was also assessed by the RDM, a live/dead bacterial fluorescence assay and plate counting. Using the RDM, the lower limit of quantitative bacterial counts in the spiked samples was determined as 3.0×10(3)(3.48log)counts mL(-1). We then used a laboratory model of a hot tub and found that the RDM could monitor the growth curve of naturally occurring heterotrophic bacteria with 1 and 2 days' delayed growth of amoeba and Legionella, respectively, and could also determine the killing curve of these bacteria by chlorination. Finally, samples with ≥3.48 or <3.48log total bacterial counts mL(-1) were tested using the RDM from 149 different hot tubs, and were found to be significantly associated with the positive or negative detection of Legionella with 95% sensitivity and 84% specificity. These findings indicated that the RDM can be used for Legionella control at bathing facilities, especially those where the effectiveness of chlorine is reduced by the presence of Fe(2+), Mn(2+), NH(4)(+), skin debris, and/or biofilms in the water. Copyright © 2011 Elsevier B.V. All rights reserved.

Tidal radii of the globular clusters M 5, M 12, M 13, M 15, M 53, NGC 5053 and NGC 5466 from automated star counts.

NASA Astrophysics Data System (ADS)

Lehmann, I.; Scholz, R.-D.

1997-04-01

We present new tidal radii for seven Galactic globular clusters using the method of automated star counts on Schmidt plates of the Tautenburg, Palomar and UK telescopes. The plates were fully scanned with the APM system in Cambridge (UK). Special account was given to a reliable background subtraction and the correction of crowding effects in the central cluster region. For the latter we used a new kind of crowding correction based on a statistical approach to the distribution of stellar images and the luminosity function of the cluster stars in the uncrowded area. The star counts were correlated with surface brightness profiles of different authors to obtain complete projected density profiles of the globular clusters. Fitting an empirical density law (King 1962) we derived the following structural parameters: tidal radius r_t_, core radius r_c_ and concentration parameter c. In the cases of NGC 5466, M 5, M 12, M 13 and M 15 we found an indication for a tidal tail around these objects (cf. Grillmair et al. 1995).

The Role of Flushing Dental Waterlines for the Removal of Microbial Contaminants - MCEARD

EPA Science Inventory

Objectives. This study was designed to determine the role of flushing dental water lines for the removal of heterotrophic plate count bacteria, Legionella spp., and free-living protozoa. Methods. Forty dental offices were surveyed in the study. An initial sample and a sample tak...

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

EPA Science Inventory

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

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

PubMed

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

2009-05-01

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

A survey of the bacteriological quality of preroasted peanut, almond, cashew, hazelnut, and brazil nut kernels received into three Australian nut-processing facilities over a period of 3 years.

PubMed

Eglezos, Sofroni; Huang, Bixing; Stuttard, Ed

2008-02-01

There is little information about bacteriological quality of preroasted kernels available in the public domain. An investigation of the bacteriological quality of preroasted peanut, almond, cashew, hazelnut, and Brazil nut kernels received into three Australian nut-processing facilities was performed over a period of 3 years. A total of 836 samples were analyzed for aerobic plate count, and 921 samples for Salmonella and Escherichia coli. The 921 samples included 653 peanut, 100 cashew, 60 almond, 60 Brazil nut, and 48 hazelnut kernels. There was no E. coli detected in any sample. Salmonella subsp. II (Fremantle) was detected in one raw almond sample. The aerobic plate count percentages of positive samples with counts above the detection level of the plating method used (100 CFU/g) for peanuts, almonds, cashews, hazelnuts, and Brazil nuts were 84, 78, 74, 50, and 45%, respectively. Of the samples containing more than this detection limit, the means were 4.5, 4.4, 3.1, 2.5, and 3.8 log CFU/g respectively. Although roasted kernel quality was not within the scope of this survey, raw microbial bioload would be expected to reduce on roasting. The bacteriological quality of preroasted peanut, almond, cashew, hazelnut, and Brazil nut kernels received into nut-processing facilities in Australia does not appear to suggest a public health concern.

Microbiological survey of a South African poultry processing plant.

PubMed

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

1995-01-01

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

Assessing the growth and recovery of Salmonella Enteritidis SE86 after sodium dichloroisocyanurate exposure

PubMed Central

Ferreira, Fernanda Stoduto; Horvath, Mariana Bandeira; Tondo, Eduardo Cesar

2013-01-01

The objective of the present study was to assess the growth and the recovery of Salmonella (S.) Enteritidis SE86 in different diluents, culture media and using different plating methods after the exposure to 200 mg/kg sodium dichloroisocyanurate (NaDCC). Before and after NaDCC exposure, SE86 was cultured at 30 °C and 7 °C in the following diluents: Peptone water (P), Saline solution (SaS), Peptone water+Saline solution (P+SaS), Peptone water+Tween 80+Lecithin+Sodium thiosulfate (P+N) and Saline solution+Tween 80+Lecithin+Sodium thiosulfate (SaS+N). The SaS diluent was chosen because it was able to maintain cells viable without growth and was further used for plating SE86 on non selective medium (Tryptic Soy Agar-TSA) and on selective media (Mannitol Lysine Crystal Violet Brilliant Green Agar-MLCB; Brilliant Green Agar-BGA; Salmonella Shigella Agar-SS and Xylose Lysine Dextrose–XLD). The Thin Agar Layer method (TAL) i.e., selective media overlayed with non selective TSA was also evaluated. Results indicated that SE86 not exposed to NaDCC was able to grow in P, P+N, SaS+N and P+SaS, but not in SaS, that was able to maintain cells viable. SE86 exposed to NaDCC demonstrated similar counts after dilution in SaS and the plating on non selective TSA, selective media MLCB, BGA, SS and XLD and on TAL media. SE86, S. Typhimurium and S. Bredeney, exposed or not exposed to NaDCC, showed no significant differences in counts on TSA, XLD and XLD overlayed with TSA, suggesting that all those media may be used to quantify NaDCC-exposed Salmonella by plating method. PMID:24516446

Fluorometric determination of the DNA concentration in municipal drinking water.

PubMed Central

McCoy, W F; Olson, B H

1985-01-01

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

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

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

Fliermans, C.B.; Dougherty, J.M.; Franck, M.M.

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.more » 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.« less

The Degree of Bacterial Contamination While Performing Spine Surgery

PubMed Central

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

2013-01-01

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

Quantitative Urine Culture Method Using a Plastic „Paddle” Containing Dual Media

PubMed Central

Craig, William A.; Kunin, Calvin M.

1972-01-01

A new dip-inoculum method for quantitative urine culture is described which utilizes a dual-chambered plastic „paddle” housing both a general purpose and differential medium. Comparative bacterial counts of 1,000 clinical specimens using the pour plate and this device were identical in 82.9% and within a factor of five in 95.6%. The „paddle” detected all but 19 of 258 specimens (92.6%) with 100,000 or greater colonies per ml. This simple, convenient method should allow more extensive use of quantitative urine culture in the diagnosis and follow-up of patients with urinary tract infections in office practice. It should not be considered as a substitute for the more definitive pour plate method or for standard methods for characterization of bacteriological species when more exact information is required. PMID:4555636

Rapid detection of total and viable Legionella pneumophila in tap water by immunomagnetic separation, double fluorescent staining and flow cytometry.

PubMed

Keserue, Hans-Anton; Baumgartner, Andreas; Felleisen, Richard; Egli, Thomas

2012-11-01

We developed a rapid detection method for Legionella pneumophila (Lp) by filtration, immunomagnetic separation, double fluorescent staining, and flow cytometry (IMS-FCM method). The method requires 120 min and can discriminate 'viable' and 'membrane-damaged' cells. The recovery is over 85% of spiked Lp SG 1 cells in 1 l of tap water and detection limits are around 50 and 15 cells per litre for total and viable Lp, respectively. The method was compared using water samples from house installations in a blind study with three environmental laboratories performing the ISO 11731 plating method. In 53% of the water samples from different taps and showers significantly higher concentrations of Lp were detected by flow cytometry. No correlation to the plate culture method was found. Since also 'viable but not culturable' (VNBC) cells are detected by our method, this result was expected. The IMS-FCM method is limited by the specificity of the used antibodies; in the presented case they target Lp serogroups 1-12. This and the fact that no Lp-containing amoebae are detected may explain why in 21% of all samples higher counts were observed using the plate culture method. Though the IMS-FCM method is not yet fit to completely displace the established plating method (ISO 11731) for routine Lp monitoring, it has major advantages to plating and can quickly provide important insights into the ecology of this pathogen in water distribution systems. © 2012 The Authors. Microbial Biotechnology © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

Application of the microbiological method DEFT/APC to detect minimally processed vegetables treated with gamma radiation

NASA Astrophysics Data System (ADS)

Araújo, M. M.; Duarte, R. C.; Silva, P. V.; Marchioni, E.; Villavicencio, A. L. C. H.

2009-07-01

Marketing of minimally processed vegetables (MPV) are gaining impetus due to its convenience, freshness and apparent health effect. However, minimal processing does not reduce pathogenic microorganisms to safe levels. Food irradiation is used to extend the shelf life and to inactivate food-borne pathogens. In combination with minimal processing it could improve safety and quality of MPV. A microbiological screening method based on the use of direct epifluorescent filter technique (DEFT) and aerobic plate count (APC) has been established for the detection of irradiated foodstuffs. The aim of this study was to evaluate the applicability of this technique in detecting MPV irradiation. Samples from retail markets were irradiated with 0.5 and 1.0 kGy using a 60Co facility. In general, with a dose increment, DEFT counts remained similar independent of the irradiation while APC counts decreased gradually. The difference of the two counts gradually increased with dose increment in all samples. It could be suggested that a DEFT/APC difference over 2.0 log would be a criteria to judge if a MPV was treated by irradiation. The DEFT/APC method could be used satisfactorily as a screening method for indicating irradiation processing.

Use of Dehydrated Agar to Estimate Microbial Water Quality for Horticulture Irrigation.

PubMed

Meador, Dustin P; Fisher, Paul R; Guy, Charles L; Harmon, Philip F; Peres, Natalia A; Teplitski, Max

2016-07-01

Petrifilms are dehydrated agar culture plates that have been used to quantify colony forming units (CFU) mL of either aerobic bacteria (Petrifilm-AC) or fungus (Petrifilm-YM), depending on substrate composition. Microbes in irrigation systems can indicate biofilm risk and potential clogging of irrigation emitters. The research objective was to compare counts on Petrifilms versus traditional, hydrated-agar plates using samples collected from recirculated irrigation waters and cultures of isolated known species. The estimated count (in CFU mL) from a recirculated irrigation sample after 7 d of incubation on Petrifilm-YM was only 5.5% of the count quantified using sabouraud dextrose agar (SDA) with chloramphenicol after 14 d. In a separate experiment with a known species, Petrifilm-YM did not successfully culture zoospores of . Isolates of viable zoospores were cultured successfully on potato-dextrose agar (PDA), with comparable counts with a vegetable juice medium supplemented with the antibiotics pimaricin, ampicillin, rifamycin, pentochloronitrobenzene and hymexazol (PARP-H). The quantification of pv. Begoniaceae on Petrifilm-AC was not significantly different ( < 0.05) than on PDA, but was lower than on Reasoner and Goldrich agar (R2A) or with a hemocytometer. The current formulation of Petrifilm-YM is unlikely to be a useful monitoring method for plant pathogens in irrigation water because of the inability to successfully culture oomycetes. However, Petrifilm-AC was an effective method to quantify bacteria and can provide an easy-to-use on-farm tool to monitor biofilm risk and microbial density. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

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

PubMed

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

2002-02-01

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

Inactivation of Escherichia coli and coliform bacteria in traditional brass and earthernware water storage vessels.

PubMed

Tandon, Puja; Chhibber, Sanjay; Reed, Robert H

2005-07-01

The detection and enumeration of indicator bacteria such as Escherichia coli is used to assess the extent of faecal contamination of drinking water. On the basis of this approach, the effectiveness of storing water contaminated with faecal indicator bacteria in brass or earthern vessels (mutkas) of the type used in rural India have been investigated. Suspensions of bacteria in sterile distilled water were maintained for up to 48 h in each vessel and enumerated by surface plate counts on nutrient agar (non-selective) and several selective coliform media at 37 degrees C either under standard aerobic conditions, or under conditions designed to neutralise reactive oxygen species (ROS), e.g. using an anaerobic cabinet to prepare plates of pre-reduced growth medium or by inclusion of sodium pyruvate in the growth medium, with incubation of aerobically-prepared plates in an anaerobic jar. The counts obtained for E. coli decreased on short-term storage in a brass mutka; counts for selective media were lower than for equivalent counts for non-selective medium, with ROS-neutralised conditions giving consistently higher counts than aerobic incubation. However, after 48 h, no bacteria were cultivable under any conditions. Similar results were obtained using water from environmental sources in the Panjab, and from rural households where brass and earthern mutkas are used for storage of drinking water, with enumeration on selective coliform media (presumptive total coliforms). In all cases results indicated that, while storage of water in a brass mutka can inactivate E. coli and coliforms over a 48 h period, standard aerobic plate counting using selective media may not be fully effective in enumerating sub-lethally damaged bacteria.

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

PubMed

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

2015-12-01

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

Project environmental microbiology as related to planetary quarantine

NASA Technical Reports Server (NTRS)

Pflug, I. J.

1974-01-01

Microbiological analyses of soil particles allow for the following conclusions: (1) there is a considerable range in the values of aerobic, mesophilic microbial counts associated with different size soil fractions; (2) as soil particle size increases, there is an increase in the mean microbial concentration per particle; (3) plate counts of aerobic, mesophilic organisms in unheated soils yielded a mean concentration of about six organisms per particle for the smallest soil fraction; (4) aerobic, mesophilic counts for sonicated particles heated at 80 C for 20 minutes yielded mean values of about two organisms per particle for the smallest particles; (5) some actinomycetes associated with the soil fractions could survive dry heat treatment at 110 C for one hour; and (6) soil particles stored under ambient laboratory conditions for 2.5 years aerobic, mesophilic plate counts which were comparable or slightly greater than the counts for more recently collected soil.

Rapid Membrane Filtration-Epifluorescent Microscopy Technique for Direct Enumeration of Bacteria in Raw Milk

PubMed Central

Pettipher, Graham L.; Mansell, Roderick; McKinnon, Charles H.; Cousins, Christina M.

1980-01-01

Membrane filtration and epifluorescent microscopy were used for the direct enumeration of bacteria in raw milk. Somatic cells were lysed by treatment with trypsin and Triton X-100 so that 2 ml of milk containing up to 5 × 106 somatic cells/ml could be filtered. The majority of the bacteria (ca. 80%) remained intact and were concentrated on the membrane. After being stained with acridine organe, the bacteria fluoresced under ultraviolet light and could easily be counted. The clump count of orange fluorescing cells on the membrane correlated well (r = 0.91) with the corresponding plate count for farm, tanker, and silo milks. Differences between counts obtained by different operators and between the membrane clump count and plate count were not significant. The technique is rapid, taking less than 25 min, inexpensive, costing less than 50 cents per sample, and is suitable for milks containing 5 × 103 to 5 × 108 bacteria per ml. Images PMID:16345515

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

NASA Astrophysics Data System (ADS)

Sams, Bruce J., III

1991-03-01

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

A quantitative PCR approach for quantification of functional genes involved in the degradation of polycyclic aromatic hydrocarbons in contaminated soils.

PubMed

Shahsavari, Esmaeil; Aburto-Medina, Arturo; Taha, Mohamed; Ball, Andrew S

2016-01-01

Polycyclic aromatic hydrocarbons (PAHs) are major pollutants globally and due to their carcinogenic and mutagenic properties their clean-up is paramount. Bioremediation or using PAH degrading microorganisms (mainly bacteria) to degrade the pollutants represents cheap, effective methods. These PAH degraders harbor functional genes which help microorganisms use PAHs as source of food and energy. Most probable number (MPN) and plate counting methods are widely used for counting PAHs degraders; however, as culture based methods only count a small fraction (<1%) of microorganisms capable of carrying out PAH degradation, the use of culture-independent methodologies is desirable.•This protocol presents a robust, rapid and sensitive qPCR method for the quantification of the functional genes involved in the degradation of PAHs in soil samples.•This protocol enables us to screen a vast number of PAH contaminated soil samples in few hours.•This protocol provides valuable information about the natural attenuation potential of contaminated soil and can be used to monitor the bioremediation process.

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

PubMed

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.

Microbial contamination in intraoral phosphor storage plates: the dilemma.

PubMed

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

2017-01-01

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

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

PubMed Central

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

2011-01-01

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

Flow cytometric bacterial cell counts challenge conventional heterotrophic plate counts for routine microbiological drinking water monitoring.

PubMed

Van Nevel, S; Koetzsch, S; Proctor, C R; Besmer, M D; Prest, E I; Vrouwenvelder, J S; Knezev, A; Boon, N; Hammes, F

2017-04-15

Drinking water utilities and researchers continue to rely on the century-old heterotrophic plate counts (HPC) method for routine assessment of general microbiological water quality. Bacterial cell counting with flow cytometry (FCM) is one of a number of alternative methods that challenge this status quo and provide an opportunity for improved water quality monitoring. After more than a decade of application in drinking water research, FCM methodology is optimised and established for routine application, supported by a considerable amount of data from multiple full-scale studies. Bacterial cell concentrations obtained by FCM enable quantification of the entire bacterial community instead of the minute fraction of cultivable bacteria detected with HPC (typically < 1% of all bacteria). FCM measurements are reproducible with relative standard deviations below 3% and can be available within 15 min of samples arriving in the laboratory. High throughput sample processing and complete automation are feasible and FCM analysis is arguably less expensive than HPC when measuring more than 15 water samples per day, depending on the laboratory and selected staining procedure(s). Moreover, many studies have shown FCM total (TCC) and intact (ICC) cell concentrations to be reliable and robust process variables, responsive to changes in the bacterial abundance and relevant for characterising and monitoring drinking water treatment and distribution systems. The purpose of this critical review is to initiate a constructive discussion on whether FCM could replace HPC in routine water quality monitoring. We argue that FCM provides a faster, more descriptive and more representative quantification of bacterial abundance in drinking water. Copyright © 2017 Elsevier Ltd. All rights reserved.

Protocols and programs for high-throughput growth and aging phenotyping in yeast.

PubMed

Jung, Paul P; Christian, Nils; Kay, Daniel P; Skupin, Alexander; Linster, Carole L

2015-01-01

In microorganisms, and more particularly in yeasts, a standard phenotyping approach consists in the analysis of fitness by growth rate determination in different conditions. One growth assay that combines high throughput with high resolution involves the generation of growth curves from 96-well plate microcultivations in thermostated and shaking plate readers. To push the throughput of this method to the next level, we have adapted it in this study to the use of 384-well plates. The values of the extracted growth parameters (lag time, doubling time and yield of biomass) correlated well between experiments carried out in 384-well plates as compared to 96-well plates or batch cultures, validating the higher-throughput approach for phenotypic screens. The method is not restricted to the use of the budding yeast Saccharomyces cerevisiae, as shown by consistent results for other species selected from the Hemiascomycete class. Furthermore, we used the 384-well plate microcultivations to develop and validate a higher-throughput assay for yeast Chronological Life Span (CLS), a parameter that is still commonly determined by a cumbersome method based on counting "Colony Forming Units". To accelerate analysis of the large datasets generated by the described growth and aging assays, we developed the freely available software tools GATHODE and CATHODE. These tools allow for semi-automatic determination of growth parameters and CLS behavior from typical plate reader output files. The described protocols and programs will increase the time- and cost-efficiency of a number of yeast-based systems genetics experiments as well as various types of screens.

Ecology and thermal inactivation of microbes in and on interplanetary space vehicle components

NASA Technical Reports Server (NTRS)

Campbell, J. E.; Reyes, A. L.; Wehby, A. J.; Crawford, R. G.; Wimsatt, J. C.; Peeler, J. T.

1974-01-01

Dry heat sterilization of spacecraft was investigated by studying the production of spore crops, and thermal inactivation of the spores, and bacillus subtillus. Spore assays were made by conventional plate count methods, and survival curves for the spores are presented. The results indicate that the inherent resistance of spores from a parent cell can be maintained.

Further investigation of relationships between membrane fluidity and ethanol tolerance in Saccharomyces cerevisiae.

PubMed

Ishmayana, Safri; Kennedy, Ursula J; Learmonth, Robert P

2017-11-27

Membrane lipid unsaturation index and membrane fluidity have been related to yeast ethanol stress tolerance in published studies, however findings have been inconsistent. In this study, viability reduction on exposure to 18% (v/v) ethanol was compared to membrane fluidity determined by laurdan generalized polarization. Furthermore, in the determination of viability reduction, we examined the effectiveness of two methods, namely total plate count and methylene violet staining. We found a strong negative correlation between ethanol tolerance and membrane fluidity, indicated by negative Pearson correlation coefficients of - 0.79, - 0.65 and - 0.69 for Saccharomyces cerevisiae strains A12, PDM and K7, respectively. We found that lower membrane fluidity leads to higher ethanol tolerance, as indicated by decreased viability reduction and higher laurdan generalized polarization in respiratory phase compared to respiro-fermentative phase cells. Total plate count better differentiated ethanol tolerance of yeast cells in different growth phases, while methylene violet staining was better to differentiate ethanol tolerance of the different yeast strains at a particular culture phase. Hence, both viability assessment methods have their own advantages and limitations, which should be considered when comparing stress tolerance in different situations.

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

PubMed Central

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

2010-01-01

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

[A comparison between 2 different methods for calculating the percentage of anaerobic microorganisms in the subgingival microbial flora].

PubMed

Petti, S; Renzini, G

1994-03-01

The percentage of anaerobic micro-organisms in the subgingival microflora represents a simple microbiological index which not only refers to the state but also the risks of periodontal health. The present study aimed to compare two different methods of calculating this index. The study was performed in 45 subjects with moderate gingivitis provoked by the previous application of dental fixtures anchored to both arches. A sample of subgingival microflora was collected from each patient at the level of the vestibular gingival sulcus of the first upper right molar. This was then vortexed, diluted and inoculated in three series of plates. It was chosen to use Walker's culture medium. The total bacterial count was evaluated by incubating the first series of plates in anaerobiosis; the anaerobic bacterial was calculated by subtracting from the total the of facultative aerobic-anaerobic micro-organisms, which in turn was obtained using two methods: the first (method AE) consisted of incubating another series of plates in aerobiosis; the second (method M) involved incubating the last series of plates in anaerobiosis, and adding metronidazole to the culture medium in a solution of 2.5 mg/l. The plates were then kept at 37 degrees C for seven days. The mean percentage of anaerobic microorganisms, given by the percentage ratio between anaerobic and total, relating to the 45 cases studied, was as follows: using method AE: 57.8 +/- 26.3%, and using method M: 40.2 +/- 27.2%. Both figures come close to that proposed and calculated using a much more sophisticated method by Slots, namely 41.5 +/- 19.2% in the event of gingivitis.(ABSTRACT TRUNCATED AT 250 WORDS)

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

PubMed

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

2016-06-07

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

Epifluorescence and atomic force microscopy: Two innovative applications for studying phage-host interactions in Lactobacillus helveticus.

PubMed

Zago, Miriam; Scaltriti, Erika; Fornasari, Maria Emanuela; Rivetti, Claudio; Grolli, Stefano; Giraffa, Giorgio; Ramoni, Roberto; Carminati, Domenico

2012-01-01

Bacteriophages attacking lactic acid bacteria (LAB) still represent a crucial problem in industrial dairy fermentations. The consequences of a phage infection against LAB can lead to fermentation delay, alteration of the product quality and, in most severe cases, the product loss. Phage particles enumeration and phage-host interactions are normally evaluated by conventional plaque count assays, but, in many cases, these methods can be unsuccessful. Bacteriophages of Lactobacillus helveticus, a LAB species widely used as dairy starter or probiotic cultures, are often unable to form lysis plaques, thus impairing their enumeration by plate assay. In this study, we used epifluorescence microscopy to enumerate L. helveticus phage particles from phage-infected cultures and Atomic Force Microscopy (AFM) to visualize both phages and bacteria during the different stages of the lytic cycle. Preliminary, we tested the sensitivity of phage counting by epifluorescence microscopy. To this end, phage particles of ΦAQ113, a lytic phage of L. helveticus isolated from a whey starter culture, were stained by SYBR Green I and enumerated by epifluorescence microscopy. Values obtained by the microscopic method were 10 times higher than plate counts, with a lowest sensitivity limit of ≥6log phage/ml. The interaction of phage ΦAQ113 with its host cell L. helveticus Lh1405 was imaged by AFM after 0, 2 and 5h from phage-host adsorption. The lytic cycle was followed by epifluorescence microscopy counting and the concomitant cell wall changes were visualized by AFM imaging. Our results showed that these two methods can be combined for a reliable phage enumeration and for studying phage and host morphology during infection processes, thus giving a complete overview of phage-host interactions in L. helveticus strains involved in dairy productions. Copyright © 2011 Elsevier B.V. All rights reserved.

A new application of a sodium deoxycholate-propidium monoazide-quantitative PCR assay for rapid and sensitive detection of viable Cronobacter sakazakii in powdered infant formula.

PubMed

Zhou, Baoqing; Chen, Bolu; Wu, Xin; Li, Fan; Yu, Pei; Aguilar, Zoraida P; Wei, Hua; Xu, Hengyi

2016-12-01

A rapid, reliable, and sensitive method for the detection of Cronobacter sakazakii, a common foodborne pathogen that may cause serious neonatal disease, has been developed. In this study, a rapid real-time quantitative PCR (qPCR) assay combined with sodium deoxycholate (SD) and propidium monoazide (PMA) was developed to detect C. sakazakii contamination in powdered infant formula (PIF). This method could eliminate the interference from dead or injured bacteria. Optimization studies indicated that SD and PMA at 0.08% (wt/vol) and 5µg/mL, respectively, were the most appropriate. In addition, qPCR, PMA-qPCR, SD-PMA-qPCR, and plate count assays were used to account for the number of viable bacteria in cell suspensions that were exposed to a 55°C water bath at different length of time. As a result, the viable number by PMA-qPCR showed significantly higher than of the number from SD-PMA-qPCR or plate counts. The number of viable bacteria was consistent between SD-PMA-qPCR and traditional plate counts, which indicated that SD treatment could eliminate the interference from dead or injured cells. Using the optimized parameters, the limit of detection with the SD-PMA-qPCR assay was 3.3×10 2 cfu/mL and 4.4×10 2 cfu/g in pure culture and in spiked PIF, respectively. A similar detection limit of 5.6×10 2 cfu/g was obtained in the presence of the Staphylococcus aureus (10 7 cfu/mL). The combined SD-PMA-qPCR assay holds promise for the rapid detection of viable C. sakazakii in PIF. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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

2018-06-01

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

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

PubMed

Kim, Shin Woong; Moon, Jongmin; Jeong, Seung-Woo; An, Youn-Joo

2018-05-01

Since the introduction of standardized nematode toxicity assays by the American Society for Testing and Materials (ASTM) and International Organization for Standardization (ISO), many studies have reported their use. Given that the currently used standardized nematode toxicity assays have certain limitations, in this study, we examined the use of a novel nematode offspring counting assay for evaluating soil ecotoxicity based on a previous soil-agar isolation method used to recover live adult nematodes. In this new assay, adult Caenorhabditis elegans were exposed to soil using a standardized toxicity assay procedure, and the resulting offspring in test soils attracted by a microbial food source in agar plates were counted. This method differs from previously used assays in terms of its endpoint, namely, the number of nematode offspring. The applicability of the bioassay was demonstrated using metal-spiked soils, which revealed metal concentration-dependent responses, and with 36 field soil samples characterized by different physicochemical properties and containing various metals. Principal component analysis revealed that texture fraction (clay, sand, and silt) and electrical conductivity values were the main factors influencing the nematode offspring counting assay, and these findings warrant further investigation. The nematode offspring counting assay is a rapid and simple process that can provide multi-directional toxicity assessment when used in conjunction with other standard methods. Copyright © 2018 Elsevier Ltd. All rights reserved.

Decontamination of a drinking water pipeline system contaminated with adenovirus and Escherichia coli utilizing peracetic acid and chlorine.

PubMed

Kauppinen, Ari; Ikonen, Jenni; Pursiainen, Anna; Pitkänen, Tarja; Miettinen, Ilkka T

2012-09-01

A contaminated drinking water distribution network can be responsible for major outbreaks of infections. In this study, two chemical decontaminants, peracetic acid (PAA) and chlorine, were used to test how a laboratory-scale pipeline system can be cleaned after simultaneous contamination with human adenovirus 40 (AdV40) and Escherichia coli. In addition, the effect of the decontaminants on biofilms was followed as heterotrophic plate counts (HPC) and total cell counts (TCC). Real-time quantitative polymerase chain reaction (qPCR) was used to determine AdV40 and plate counting was used to enumerate E. coli. PAA and chlorine proved to be effective decontaminants since they decreased the levels of AdV40 and E. coli to below method detection limits in both water and biofilms. However, without decontamination, AdV40 remained present in the pipelines for up to 4 days. In contrast, the concentration of cultivable E. coli decreased rapidly in the control pipelines, implying that E. coli may be an inadequate indicator for the presence of viral pathogens. Biofilms responded to the decontaminants by decreased HPCs while TCC remained stable. This indicates that the mechanism of pipeline decontamination by chlorine and PAA is inactivation rather than physical removal of microbes.

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

PubMed

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

1993-01-01

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

A novel quantitative reverse-transcription PCR (qRT-PCR) for the enumeration of total bacteria, using meat micro-flora as a model.

PubMed

Dolan, Anthony; Burgess, Catherine M; Barry, Thomas B; Fanning, Seamus; Duffy, Geraldine

2009-04-01

A sensitive quantitative reverse-transcription PCR (qRT-PCR) method was developed for enumeration of total bacteria. Using two sets of primers separately to target the ribonuclease-P (RNase P) RNA transcripts of gram positive and gram negative bacteria. Standard curves were generated using SYBR Green I kits for the LightCycler 2.0 instrument (Roche Diagnostics) to allow quantification of mixed microflora in liquid media. RNA standards were used and extracted from known cell equivalents and subsequently converted to cDNA for the construction of standard curves. The number of mixed bacteria in culture was determined by qRT-PCR, and the results correlated (r(2)=0.88, rsd=0.466) with the total viable count over the range from approx. Log(10) 3 to approx. Log(10) 7 CFU ml(-1). The rapid nature of this assay (8 h) and its potential as an alternative method to the standard plate count method to predict total viable counts and shelf life are discussed.

Automatic counting and classification of bacterial colonies using hyperspectral imaging

USDA-ARS?s Scientific Manuscript database

Detection and counting of bacterial colonies on agar plates is a routine microbiology practice to get a rough estimate of the number of viable cells in a sample. There have been a variety of different automatic colony counting systems and software algorithms mainly based on color or gray-scale pictu...

Efficient method for the detection of microbially-produced antibacterial substances from food systems.

PubMed

Morgan, S M; Hickey, R; Ross, R P; Hill, C

2000-07-01

A novel method for the isolation of microbially-derived inhibitory substances from food sources was developed. The method involves an enrichment step coupled to a killing assay which is initially carried out in multiwell plates. The technique has advantages in that large numbers of samples can be tested in parallel. The assay can be completed in less than 60 h and is more sensitive than direct plating due to the enrichment step. This novel screening approach was compared with the standard direct plating approach in an effort to identify the antimicrobial potential of a number of Kefir grains. Kefir grains were incubated in 10% reconstituted skim milk for 20 h at 32 degrees C to enable production of any potential biopreservatives. Following overnight incubation, fermentates were aliquoted into multi-well plates and a known number of indicator cells was added to each well. The fermentates were incubated for a further 20 h and counts were carried out to determine whether a reduction in indicator cell numbers had occurred. A reduction in cell-forming units indicated the presence of an inhibitory substance and these inhibitory fermentates were selected for further investigation. Using the protocol outlined, Kefir fermentates capable of inhibiting Listeria innocua DPC1770 and Escherichia coli O157:H45 were identified.

Optimization, validation, and application of a real-time PCR protocol for quantification of viable bacterial cells in municipal sewage sludge and biosolids using reporter genes and Escherichia coli.

PubMed

van Frankenhuyzen, Jessica K; Trevors, Jack T; Flemming, Cecily A; Lee, Hung; Habash, Marc B

2013-11-01

Biosolids result from treatment of sewage sludge to meet jurisdictional standards, including pathogen reduction. Once government regulations are met, materials can be applied to agricultural lands. Culture-based methods are used to enumerate pathogen indicator microorganisms but may underestimate cell densities, which is partly due to bacteria existing in a viable but non-culturable physiological state. Viable indicators can also be quantified by realtime polymerase chain reaction (qPCR) used with propidium monoazide (PMA), a dye that inhibits amplification of DNA found extracellularly or in dead cells. The objectives of this study were to test an optimized PMA-qPCR method for viable pathogen detection in wastewater solids and to validate it by comparing results to data obtained by conventional plating. Reporter genes from genetically marked Pseudomonas sp. UG14Lr and Agrobacterium tumefaciens 542 cells were spiked into samples of primary sludge, and anaerobically digested and Lystek-treated biosolids as cell-free DNA, dead cells, viable cells, and mixtures of live and dead cells, followed by DNA extraction with and without PMA, and qPCR. The protocol was then used for Escherichia coli quantification in the three matrices, and results compared to plate counts. PMA-qPCR selectively detected viable cells, while inhibiting signals from cell-free DNA and DNA found in membrane-compromised cells. PMA-qPCR detected 0.5-1 log unit more viable E. coli cells in both primary solids and dewatered biosolids than plate counts. No viable E. coli was found in Lystek-treated biosolids. These data suggest PMA-qPCR may more accurately estimate pathogen cell numbers than traditional culture methods.

Antimicrobial susceptibility testing of Mycobacterium tuberculosis complex for first and second line drugs by broth dilution in a microtiter plate format.

PubMed

Hall, Leslie; Jude, Kurt P; Clark, Shirley L; Wengenack, Nancy L

2011-06-24

The rapid detection of antimicrobial resistance is important in the effort to control the increase in resistant Mycobacterium tuberculosis (Mtb). Antimicrobial susceptibility testing (AST) of Mtb has traditionally been performed by the agar method of proportion or by macrobroth testing on an instrument such as the BACTEC (Becton Dickinson, Sparks, MD), VersaTREK (TREK Diagnostics, Cleveland, OH) or BacT/ALERT (bioMérieux, Hazelwood, MO). The agar proportion method, while considered the "gold" standard of AST, is labor intensive and requires calculation of resistance by performing colony counts on drug-containing agar as compared to drug-free agar. If there is ≥1% growth on the drug-containing medium as compared to drug-free medium, the organism is considered resistant to that drug. The macrobroth methods require instrumentation and test break point ("critical") drug concentrations for the first line drugs (isoniazid, ethambutol, rifampin, and pyrazinamide). The method described here is commercially available in a 96 well microtiter plate format [MYCOTB (TREK Diagnostics)] and contains increasing concentrations of 12 antimicrobials used for treatment of tuberculosis including both first (isoniazid, rifampin, ethambutol) and second line drugs (amikacin, cycloserine, ethionamide, kanamycin, moxifloxacin, ofloxacin, para-aminosalicylic acid, rifabutin, and streptomycin). Pyrazinamide, a first line drug, is not included in the microtiter plate due to its need for acidic test conditions. Advantages of the microtiter system include both ease of set up and faster turn around time (14 days) compared with traditional agar proportion (21 days). In addition, the plate can be set up from inoculum prepared using either broth or solid medium. Since the microtiter plate format is new and since Mtb presents unique safety challenges in the laboratory, this protocol will describe how to safely setup, incubate and read the microtiter plate.

Low noise and conductively cooled microchannel plates

NASA Technical Reports Server (NTRS)

Feller, W. B.

1990-01-01

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

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

NASA Technical Reports Server (NTRS)

Timothy, J. G.

1975-01-01

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

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

NASA Technical Reports Server (NTRS)

Timothy, J. G.

1976-01-01

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

Smartphone-based rapid quantification of viable bacteria by single-cell microdroplet turbidity imaging.

PubMed

Cui, Xiaonan; Ren, Lihui; Shan, Yufei; Wang, Xixian; Yang, Zhenlong; Li, Chunyu; Xu, Jian; Ma, Bo

2018-05-18

Standard plate count (SPC) has been recognized as the golden standard for the quantification of viable bacteria. However, SPC usually takes one to several days to grow individual cells into a visible colony, which greatly hampers its application in rapid bacteria enumeration. Here we present a microdroplet turbidity imaging based digital standard plate count (dSPC) method to overcome this hurdle. Instead of cultivating on agar plates, bacteria are encapsulated in monodisperse microdroplets for single-cell cultivation. Proliferation of the encapsulated bacterial cell produced a detectable change in microdroplet turbidity, which allowed, after just a few bacterial doubling cycles (i.e., a few hours), enumeration of viable bacteria by visible-light imaging. Furthermore, a dSPC platform integrating a power-free droplet generator with smartphone-based turbidity imaging was established. As proof-of-concept demonstrations, a series of Gram-negative bacteria (Escherichia coli) and Gram-positive bacteria (Bacillus subtilis) samples were quantified via the smartphone dSPC accurately within 6 hours, representing a detection sensitivity of 100 CFU ml-1 and at least 3 times faster. In addition, Enterobacter sakazakii (E. sakazakii) in infant milk powder as a real sample was enumerated within 6 hours, in contrast to the 24 hours needed in traditional SPC. Results with high accuracy and reproducibility were achieved, with no difference in counts found between dSPC and SPC. By enabling label-free, rapid, portable and low-cost enumeration and cultivation of viable bacteria onsite, smartphone dSPC forms the basis for a temporally and geographically trackable network for surveying live microbes globally where every citizen with a cellphone can contribute anytime and anywhere.

A simple and highly repeatable viral plaque assay for enterovirus 71.

PubMed

Yin, Yingxian; Xu, Yi; Ou, Zhiying; Su, Ling; Xia, Huimin

2015-04-01

The classic plaque assay is a method for counting infectious viral particles, however its complexity limits its use in a variety of virological experiments. To simplify the operation and to improve the repeatability, we employed an improved plaque assay procedure based on Avicel to make the whole experiment easier and optimize the results on a model of Vero cells infection with Enterovirus 71(EV71). Clear plaques visible to the naked eyes can be formed on a 24-well plate or a 96-well plate without immunostaining. Following further improvement, this plaque assay procedure could be applied to other viruses, being both simple and repeatable. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

VizieR Online Data Catalog: Tidal radii of 7 globular clusters (Lehmann+ 1997)

NASA Astrophysics Data System (ADS)

Lehmann, I.; Scholz, R.-D.

1998-02-01

We present new tidal radii for seven Galactic globular clusters using the method of automated star counts on Schmidt plates of the Tautenburg, Palomar and UK telescopes. The plates were fully scanned with the APM system in Cambridge (UK). Special account was given to a reliable background subtraction and the correction of crowding effects in the central cluster region. For the latter we used a new kind of crowding correction based on a statistical approach to the distribution of stellar images and the luminosity function of the cluster stars in the uncrowded area. The star counts were correlated with surface brightness profiles of different authors to obtain complete projected density profiles of the globular clusters. Fitting an empirical density law (King 1962AJ.....67..471K) we derived the following structural parameters: tidal radius rt, core radius rc and concentration parameter c. In the cases of NGC 5466, M 5, M 12, M 13 and M 15 we found an indication for a tidal tail around these objects (cf. Grillmair et al., 1995AJ....109.2553G). (1 data file).

Tolerance of Chemoorganotrophic Bioleaching Microorganisms to Heavy Metal and Alkaline Stresses

PubMed Central

Monballiu, Annick; Cardon, Nele; Tri Nguyen, Minh; Cornelly, Christel; Meesschaert, Boudewijn; Chiang, Yi Wai

2015-01-01

The bioleaching potential of the bacterium Bacillus mucilaginosus and the fungus Aspergillus niger towards industrial residues was investigated by assessing their response towards various heavy metals (including arsenic, cadmium, cobalt, chromium, nickel, lead, and zinc) and elevated pH. The plate diffusion method was performed for each metal to determine the toxicity effect. Liquid batch cultures were set up for more quantitative evaluation as well as for studying the influence of basicity. Growth curves were prepared using bacterial/fungal growth counting techniques such as plate counting, optical density measurement, and dry biomass determination. Cadmium, nickel, and arsenite had a negative influence on the growth of B. mucilaginosus, whereas A. niger was sensitive to cadmium and arsenate. However, it was shown that growth recovered when microorganisms cultured in the presence of these metals were inoculated onto metal-free medium. Based on the findings of the bacteriostatic/fungistatic effect of the metals and the adaptability of the microorganisms to fairly elevated pH values, it is concluded that both strains have potential applicability for further research concerning bioleaching of alkaline waste materials. PMID:26236176

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

PubMed

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

2014-03-01

Clinical mastitis is one of the most common and expensive diseases of dairy cattle. To make an informed treatment decision, it is important to know the causative pathogen. However, no detection of bacterial growth can be made in approximately 30% of all clinical cases of mastitis. Before selecting the treatment regimen, it is important to know whether the mastitis-causing pathogen (MCP) is Gram-positive or Gram-negative. The aim of this field study was to investigate whether using two 3M Petrifilm™ products on-farm (which conveys a higher degree of sample freshness but also bears a higher risk for contamination than working in a lab) as 24-h rapid diagnostic of clinical mastitis achieved results that were comparable to the conventional microbiological diagnostic method. AerobicCount (AC)-Petrifilm™ and ColiformCount (CC)-Petrifilm™ were used to identify the total bacterial counts and Gram-negative bacteria in samples from clinical mastitis cases, respectively. Missing growth on both plates was classified as no bacterial detection. Growth only on the AC-Petrifilm™ was assessed as Gram-positive, and growth on both Petrifilm™ plates was assessed as Gram-negative bacterial growth. Additionally, milk samples were analysed by conventional microbiological diagnostic method on aesculin blood agar as a reference method. Overall, 616 samples from clinical mastitis cases were analysed. Using the reference method, Gram-positive and Gram-negative bacteria, mixed bacterial growth, contaminated samples and yeast were determined in 32.6%, 20.0%, 2.5%, 14.1% and 1.1% of the samples, respectively. In 29.7% of the samples, microbiological growth could not be identified. Using the Petrifilm™ concept, bacterial growth was detected in 59% of the culture-negative samples. The sensitivity of the Petrifilm™ for Gram-positive and Gram-negative MCP was 85.2% and 89.9%, respectively. The specificity was 75.4% for Gram-positive and 88.4% for Gram-negative MCP. For the culture-negative samples, sensitivity was 41.0% and specificity was 91.0%. The results indicate that the Petrifilm™ concept is suitable for therapeutic decision-making at the farm level or in veterinary practice. As this concept does not allow any statement about the genus or species of microorganisms, relevant MCP should be assessed periodically at the herd level with conventional microbiological diagnostics. Copyright © 2014 Elsevier B.V. All rights reserved.

Shelf life prediction of canned fried-rice using accelerated shelf life testing (ASLT) arrhenius method

NASA Astrophysics Data System (ADS)

Kurniadi, M.; Bintang, R.; Kusumaningrum, A.; Nursiwi, A.; Nurhikmat, A.; Susanto, A.; Angwar, M.; Triwiyono; Frediansyah, A.

2017-12-01

Research on shelf-life prediction of canned fried rice using Accelerated Shelf-life Test (ASLT) of Arrhenius model has been conducted. The aim of this research to predict shelf life of canned-fried rice products. Lethality value of 121°C for 15 and 20 minutes and Total Plate count methods are used to determine time and temperatures of sterilization process.Various storage temperatures of ASLT Arrhenius method were 35, 45 and 55°C during 35days. Rancidity is one of the derivation quality of canned fried rice. In this research, sample of canned fried rice is tested using rancidity value (TBA). TBA value was used as parameter which be measured once a week periodically. The use of can for fried rice without any chemical preservative is one of the advantage of the product, additionaly the use of physicalproperties such as temperature and pressure during its process can extend the shelf life and reduce the microbial contamination. The same research has never done before for fried rice as ready to eat meal. The result showed that the optimum conditions of sterilization process were 121°C,15 minutes with total plate count number of 9,3 × 101 CFU/ml. Lethality value of canned fried rice at 121°C,15 minutes was 3.63 minutes. The calculated Shelf-life of canned fried rice using Accelerated Shelf-life Test (ASLT) of Arrhenius method was 10.3 months.

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

PubMed

Quintana, Gabriel; Gerbino, Esteban; Gómez-Zavaglia, Andrea

2017-01-01

Okara is a nutritionally valuable by-product produced in large quantities as result of soymilk elaboration. This work proposes its use as both culture and dehydration medium during freeze-drying, spray-drying, and storage of Lactobacillus plantarum CIDCA 83114. Whole and defatted okara were employed as culture media for L. plantarum CIDCA 83114. The growth kinetics were followed by plate counting and compared with those of bacteria grown in MRS broth (control). No significant differences in plate counting were observed in the three media. The fatty acid composition of bacteria grown in whole and defatted okara showed a noticeable increase in the unsaturated/saturated (U/S) fatty acid ratio, with regard to bacteria grown in MRS. This change was mainly due to the increase in polyunsaturated fatty acids, namely C18:2. For dehydration assays, cultures in the stationary phase were neutralized and freeze-dried (with or without the addition of 250 mM sucrose) or spray-dried. Bacteria were plate counted immediately after freeze-drying or spray-drying and during storage at 4°C for 90 days. Freeze-drying in whole okara conducted to the highest bacterial recovery. Regarding storage, spray-dried bacteria previously grown in whole and defatted okara showed higher plate counts than those grown in MRS. On the contrary, freeze-dried bacteria previously grown in all the three culture media were those with the lowest plate counts. The addition of sucrose to the dehydration media improved their recovery. The higher recovery of microorganisms grown in okara after freeze-drying and spray-drying processes and during storage was ascribed to both the presence of fiber and proteins in the dehydration media, and the increase in U/S fatty acids ratio in bacterial membranes. The obtained results support for the first time the use of okara as an innovative matrix to deliver L. plantarum . Considering that okara is an agro-waste obtained in large quantities, these results represent an innovative strategy to add it value, providing a symbiotic ingredient with promising industrial applications in the development of novel functional foods and feeds.

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

PubMed Central

Quintana, Gabriel; Gerbino, Esteban; Gómez-Zavaglia, Andrea

2017-01-01

Okara is a nutritionally valuable by-product produced in large quantities as result of soymilk elaboration. This work proposes its use as both culture and dehydration medium during freeze-drying, spray-drying, and storage of Lactobacillus plantarum CIDCA 83114. Whole and defatted okara were employed as culture media for L. plantarum CIDCA 83114. The growth kinetics were followed by plate counting and compared with those of bacteria grown in MRS broth (control). No significant differences in plate counting were observed in the three media. The fatty acid composition of bacteria grown in whole and defatted okara showed a noticeable increase in the unsaturated/saturated (U/S) fatty acid ratio, with regard to bacteria grown in MRS. This change was mainly due to the increase in polyunsaturated fatty acids, namely C18:2. For dehydration assays, cultures in the stationary phase were neutralized and freeze-dried (with or without the addition of 250 mM sucrose) or spray-dried. Bacteria were plate counted immediately after freeze-drying or spray-drying and during storage at 4°C for 90 days. Freeze-drying in whole okara conducted to the highest bacterial recovery. Regarding storage, spray-dried bacteria previously grown in whole and defatted okara showed higher plate counts than those grown in MRS. On the contrary, freeze-dried bacteria previously grown in all the three culture media were those with the lowest plate counts. The addition of sucrose to the dehydration media improved their recovery. The higher recovery of microorganisms grown in okara after freeze-drying and spray-drying processes and during storage was ascribed to both the presence of fiber and proteins in the dehydration media, and the increase in U/S fatty acids ratio in bacterial membranes. The obtained results support for the first time the use of okara as an innovative matrix to deliver L. plantarum. Considering that okara is an agro-waste obtained in large quantities, these results represent an innovative strategy to add it value, providing a symbiotic ingredient with promising industrial applications in the development of novel functional foods and feeds. PMID:28446905

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

NASA Astrophysics Data System (ADS)

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

2015-07-01

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

APPARENT BIAS IN RIVER WATER INOCULUM FOLLOWING CENTRIFUGATION

EPA Science Inventory

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

Predicting the growth situation of Pseudomonas aeruginosa on agar plates and meat stuffs using gas sensors

PubMed Central

Gu, Xinzhe; Sun, Ye; Tu, Kang; Dong, Qingli; Pan, Leiqing

2016-01-01

A rapid method of predicting the growing situation of Pseudomonas aeruginosa is presented. Gas sensors were used to acquire volatile compounds generated by P. aeruginosa on agar plates and meat stuffs. Then, optimal sensors were selected to simulate P. aeruginosa growth using modified Logistic and Gompertz equations by odor changes. The results showed that the responses of S8 or S10 yielded high coefficients of determination (R2) of 0.89–0.99 and low root mean square errors (RMSE) of 0.06–0.17 for P. aeruginosa growth, fitting the models on the agar plate. The responses of S9, S4 and the first principal component of 10 sensors fit well with the growth of P. aeruginosa inoculated in meat stored at 4 °C and 20 °C, with R2 of 0.73–0.96 and RMSE of 0.25–1.38. The correlation coefficients between the fitting models, as measured by electronic nose responses, and the colony counts of P. aeruginosa were high, ranging from 0.882 to 0.996 for both plate and meat samples. Also, gas chromatography–mass spectrometry results indicated the presence of specific volatiles of P. aeruginosa on agar plates. This work demonstrated an acceptable feasibility of using gas sensors—a rapid, easy and nondestructive method for predicting P. aeruginosa growth. PMID:27941841

Hybrid Ion-Detector/Data-Acquisition System for a TOF-MS

NASA Technical Reports Server (NTRS)

Burton, William D., Jr.; Schultz, J. Albert; Vaughn, Valentine; McCully, Michael; Ulrich, Steven; Egan, Thomas F.

2006-01-01

A modified ion-detector/data-acquisition system has been devised to increase the dynamic range of a time-of-flight mass spectrometer (TOF-MS) that, previously, included a microchannel-plate detector and a data-acquisition system based on counting pulses and time-tagging them by use of a time-to-digital converter (TDC). The dynamic range of the TOF-MS was limited by saturation of the microchannel plate detector, which can handle no more than a few million counts per second. The modified system includes (1) a combined microchannel plate/discrete ion multiplier and (2) a hybrid data-acquisition system that simultaneously performs analog current or voltage measurements and multianode single-ion-pulse-counting time-of-flight measurements to extend the dynamic range of a TDC into the regime in which a mass peak comprises multiple ions arriving simultaneously at the detector. The multianode data are used to determine, in real time, whether the detector is saturated. When saturation is detected, the data-acquisition system selectively enables circuitry that simultaneously determines the ion-peak intensity by measuring the time profile of the analog current or voltage detector-output signal.

Clinical aspects of Candida species carriage in saliva of xerotomic subjects.

PubMed

Torres, S R; Peixoto, C B; Caldas, D M; Silva, E B; Magalhães, F A C; Uzeda, M; Nucci, M

2003-10-01

In order to investigate the clinical factors that might influence the diversity and the degree of Candida species carriage in saliva, we conducted a cross-sectional study with 133 patients with complaints of xerostomia. Anamnesis, oral examination and collection of chewing-stimulated whole saliva were performed. The samples of saliva were kept refrigerated until they were plated onto CHROMagar Candida; cfu were counted and Candida species were identified by standard methods. There was a high prevalence of mixed Candida colonization. No relationship was found between total Candida cfu counts and variables like gender, age, place of origin, underlying diseases, exposure to medications (except antibiotics), daily habits and salivary flow rates. Oral candidiasis, antibiotic exposure and dental prosthesis wearing were associated with relatively high Candida counts in saliva. Low salivary flow rates predisposed to intense colonization by C. albicans and C. parapsilosis.

Microbial and Chemical Shelf-Life of Vacuum Steam-Pasteurized Whole Flaxseed and Milled Flaxseed.

PubMed

Shah, Manoj; Eklund, Bridget; Conde Lima, Luiz Gustavo; Bergholz, Teresa; Hall, Clifford

2018-02-01

Flaxseed is an oilseed with many health benefits. Flaxseed may be consumed raw or in processed form. In the raw form, there is a potential for microbial contamination. Several pasteurization methods have been used to reduce microbial contamination. However, such treatments may affect chemical properties of foods. In this study, vacuum steam-pasteurization was conducted on whole flaxseed and milled flaxseed using 4 different conditions (3 min at 75 °C, 3 min at 90 °C, 9 min at 90 °C, and 3 min at 105 °C). Microbial and chemical shelf-life was monitored for 28 wk (36 wk for aerobic plate counts). Significant reduction (P < 0.05) in microbial counts (total aerobic plate counts, and yeast and mold counts) occurred after pasteurization and during storage of both whole flaxseed and milled flaxseed. Although both the moisture content and a w increased after pasteurization, they were similar to the unpasteurized samples during storage. Peroxide value, free fatty acid, headspace volatiles, fatty acid profiles, oil content, and secoisolariciresinol diglucoside (SDG) content were chemical indices measured. Only small changes were observed in the chemical indices after vacuum steam-pasteurization for both pasteurized whole flaxseed and milled flaxseed as compared to the unpasteurized flaxseed at most instances. Vacuum steam-pasteurization can be used as a safe alternative for the microbial reduction of low-moisture products, such as flaxseed, without significantly affecting chemical stability. Vacuum steam-pasteurization can be effectively used for the treatment of whole flaxseed and milled flaxseed to reduce spoilage microorganisms, such as total aerobes and yeasts and molds. In addition, this pasteurization method had minimal effects on several chemical shelf-life parameters with positive impact on SDG of the processed flaxseed. © 2018 Institute of Food Technologists®.

Sensitive detection of Escherichia coli O157:H7 in food and water by immunomagnetic separation and solid-phase laser cytometry

NASA Technical Reports Server (NTRS)

Pyle, B. H.; Broadaway, S. C.; McFeters, G. A.

1999-01-01

Rapid, direct methods are needed to assess active bacterial populations in water and foods. Our objective was to determine the efficiency of bacterial detection by immunomagnetic separation (IMS) and the compatibility of IMS with cyanoditolyl tetrazolium chloride (CTC) incubation to determine respiratory activity, using the pathogen Escherichia coli O157:H7. Counterstaining with a specific fluorescein-conjugated anti-O157 antibody (FAb) following CTC incubation was used to allow confirmation and visualization of bacteria by epifluorescence microscopy. Broth-grown E. coli O157:H7 was used to inoculate fresh ground beef (<17% fat), sterile 0.1% peptone, or water. Inoculated meat was diluted and homogenized in a stomacher and then incubated with paramagnetic beads coated with anti-O157 specific antibody. After IMS, cells with magnetic beads attached were stained with CTC and then an anti-O157 antibody-fluorescein isothiocyanate conjugate and filtered for microscopic enumeration or solid-phase laser cytometry. Enumeration by laser scanning permitted detection of ca. 10 CFU/g of ground beef or <10 CFU/ml of liquid sample. With inoculated meat, the regression results for log-transformed respiring FAb-positive counts of cells recovered on beads versus sorbitol-negative plate counts in the inoculum were as follows: intercept = 1.06, slope = 0.89, and r2 = 0. 95 (n = 13). The corresponding results for inoculated peptone were as follows: intercept = 0.67, slope = 0.88, and r2 = 0.98 (n = 24). Recovery of target bacteria on beads by the IMS-CTC-FAb method, compared with recovery by sorbitol MacConkey agar plating, yielded greater numbers (beef, 6.0 times; peptone, 3.0 times; water, 2.4 times). Thus, within 5 to 7 h, the IMS-CTC-FAb method detected greater numbers of E. coli O157 cells than were detected by plating. The results show that the IMS-CTC-FAb technique with enumeration by either fluorescence microscopy or solid-phase laser scanning cytometry gave results that compared favorably with plating following IMS.

Comparison of Standard Culture-Based Method to Culture-Independent Method for Evaluation of Hygiene Effects on the Hand Microbiome

PubMed Central

Leff, J.; Henley, J.; Tittl, J.; De Nardo, E.; Butler, M.; Griggs, R.; Fierer, N.

2017-01-01

ABSTRACT Hands play a critical role in the transmission of microbiota on one’s own body, between individuals, and on environmental surfaces. Effectively measuring the composition of the hand microbiome is important to hand hygiene science, which has implications for human health. Hand hygiene products are evaluated using standard culture-based methods, but standard test methods for culture-independent microbiome characterization are lacking. We sampled the hands of 50 participants using swab-based and glove-based methods prior to and following four hand hygiene treatments (using a nonantimicrobial hand wash, alcohol-based hand sanitizer [ABHS], a 70% ethanol solution, or tap water). We compared results among culture plate counts, 16S rRNA gene sequencing of DNA extracted directly from hands, and sequencing of DNA extracted from culture plates. Glove-based sampling yielded higher numbers of unique operational taxonomic units (OTUs) but had less diversity in bacterial community composition than swab-based sampling. We detected treatment-induced changes in diversity only by using swab-based samples (P < 0.001); we were unable to detect changes with glove-based samples. Bacterial cell counts significantly decreased with use of the ABHS (P < 0.05) and ethanol control (P < 0.05). Skin hydration at baseline correlated with bacterial abundances, bacterial community composition, pH, and redness across subjects. The importance of the method choice was substantial. These findings are important to ensure improvement of hand hygiene industry methods and for future hand microbiome studies. On the basis of our results and previously published studies, we propose recommendations for best practices in hand microbiome research. PMID:28351915

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

PubMed

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

2017-10-01

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

Comparing Diagnostic Accuracy of Kato-Katz, Koga Agar Plate, Ether-Concentration, and FLOTAC for Schistosoma mansoni and Soil-Transmitted Helminths

PubMed Central

Glinz, Dominik; Silué, Kigbafori D.; Knopp, Stefanie; Lohourignon, Laurent K.; Yao, Kouassi P.; Steinmann, Peter; Rinaldi, Laura; Cringoli, Giuseppe; N'Goran, Eliézer K.; Utzinger, Jürg

2010-01-01

Background Infections with schistosomes and soil-transmitted helminths exert a considerable yet underappreciated economic and public health burden on afflicted populations. Accurate diagnosis is crucial for patient management, drug efficacy evaluations, and monitoring of large-scale community-based control programs. Methods/Principal Findings The diagnostic accuracy of four copromicroscopic techniques (i.e., Kato-Katz, Koga agar plate, ether-concentration, and FLOTAC) for the detection of Schistosoma mansoni and soil-transmitted helminth eggs was compared using stool samples from 112 school children in Côte d'Ivoire. Combined results of all four methods served as a diagnostic ‘gold’ standard and revealed prevalences of S. mansoni, hookworm, Trichuris trichiura, Strongyloides stercoralis and Ascaris lumbricoides of 83.0%, 55.4%, 40.2%, 33.9% and 28.6%, respectively. A single FLOTAC from stool samples preserved in sodium acetate-acetic acid-formalin for 30 or 83 days showed a higher sensitivity for S. mansoni diagnosis (91.4%) than the ether-concentration method on stool samples preserved for 40 days (85.0%) or triplicate Kato-Katz using fresh stool samples (77.4%). Moreover, a single FLOTAC detected hookworm, A. lumbricoides and T. trichiura infections with a higher sensitivity than any of the other methods used, but resulted in lower egg counts. The Koga agar plate method was the most accurate diagnostic assay for S. stercoralis. Conclusion/Significance We have shown that the FLOTAC method holds promise for the diagnosis of S. mansoni. Moreover, our study confirms that FLOTAC is a sensitive technique for detection of common soil-transmitted helminths. For the diagnosis of S. stercoralis, the Koga agar plate method remains the method of choice. PMID:20651931

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

PubMed

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

2007-12-01

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

Attachment of Asaia bogorensis Originating in Fruit-Flavored Water to Packaging Materials

PubMed Central

Otlewska, Anna; Antolak, Hubert

2014-01-01

The objective of this study was to investigate the adhesion of isolated spoilage bacteria to packaging materials used in the food industry. Microorganisms were isolated from commercial fruit-flavored mineral water in plastic bottles with flocks as a visual defect. The Gram-negative rods were identified using the molecular method through the amplification of a partial region of the 16S rRNA gene. Based on the sequence identity (99.6%) between the spoilage organism and a reference strain deposited in GenBank, the spoilage isolate was identified as Asaia bgorensis. Experiments on bacterial adhesion were conducted using plates made of glass and polystyrene (packaging materials commonly used in the beverage industry). Cell adhesion ability was determined using luminometry, plate count, and the microscopic method. The strain of A. bogorensis was characterized by strong adhesion properties which were dependent on the surface type, with the highest cell adhesion detected on polystyrene. PMID:25295262

Quality and shelf life evaluation of fermented sausages of buffalo meat with different levels of heart and fat.

PubMed

Ahmad, S; Srivastava, P K

2007-04-01

Investigations were carried to study the effect of heart incorporation (0%, 15% and 20%) and increasing levels of fat (20% and 25%) on physicochemical (pH, moisture content and thiobarbituric acid, TBA number) and microbiological (total plate count and yeast and mold count) quality and shelf life of semi dry sausages of buffalo meat during refrigerated storage (4°C). Different levels of fat significantly (p<0.05) increased the pH of the sausage samples. However different levels of heart incorporation did not significantly (p<0.05) affect pH, moisture content and TBA number of sausage samples. Fresh samples had pH, moisture content and TBA number in the range of 5.15-5.28, 42.4-47.4% and 0.073-0.134 respectively. Refrigerated storage significantly (p<0.05) increased TBA number of control samples while storage did not significantly (p<0.05) increase the TBA number of sodium ascorbate (SA) treated samples. Total plate counts of twelve sausage samples were f under the TFTC (too few to count) limit at the initial stage. Incorporation of different levels of heart and also increasing levels of fat did not significantly (p<0.05) increase the log TPC/g values. Yeast and molds were not detected in twelve samples of semi dry fermented sausages in their fresh condition. Storage revealed that there was a consistent decrease in pH, and moisture content. Refrigerated storage significantly (p<0.05) reduced both pH and moisture contents. TBA number and total plate counts and yeast and mold counts of controls were found to increase significantly (p<0.05) during refrigerated storage. However, in SA treated sausage, only TPC and yeast and mold count significantly (p<0.05) increased during refrigerated storage. Shelf life of the sausages was found to be 60 days under refrigerated storage (4°C).

Intercondylar humerus fracture- parallel plating and its results.

PubMed

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

2015-01-01

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

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

PubMed

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

2009-08-01

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

The supercontinuum laser as a flexible source for quasi-steady state and time resolved fluorescence studies

NASA Astrophysics Data System (ADS)

Fenske, Roger; Näther, Dirk U.; Dennis, Richard B.; Smith, S. Desmond

2010-02-01

Commercial Fluorescence Lifetime Spectrometers have long suffered from the lack of a simple, compact and relatively inexpensive broad spectral band light source that can be flexibly employed for both quasi-steady state and time resolved measurements (using Time Correlated Single Photon Counting [TCSPC]). This paper reports the integration of an optically pumped photonic crystal fibre, supercontinuum source1 (Fianium model SC400PP) as a light source in Fluorescence Lifetime Spectrometers (Edinburgh Instruments FLS920 and Lifespec II), with single photon counting detectors (micro-channel plate photomultiplier and a near-infrared photomultiplier) covering the UV to NIR range. An innovative method of spectral selection of the supercontinuum source involving wedge interference filters is also discussed.

Bacteriology of Dehydrated Space Foods 1

PubMed Central

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

1971-01-01

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

[BIOMECHANICAL COMPARATIVE STUDY ON FOUR INTERNAL FIXATIONS FOR ACETABULAR FRACTURES IN QUADRILATERAL AREA].

PubMed

Wang, Lei; Wu, Xiaobo; Qi, Wei; Wang, Yongbin; He, Quanjie; Xu, Fengsong; Liu, Hongyang

2015-10-01

To compare the biomechanical difference of 4 kinds of internal fixations for acetabular fracture in quadrilateral area. The transverse fracture models were created in 16 hemipelves specimens from 8 adult males, and were randomly divided into 4 groups according to different internal fixation methods (n = 4): infrapectineal buttress reconstruction plate (group A), infrapectineal buttress locking reconstruction plate (group B), reconstruction plate combined with trans-plate quadrilateral screws (group C), and anterior reconstruction plate-lag screw (group D). Then the horizontal displacement, longitudinal displacement of fractures, and axial stiffness were measured and counted to compare the stability after continuous vertical loading. Under the same loading, the horizontal and longitudinal displacements of groups A, B, C, and D were decreased gradually; when the loading reached 1 800 N, the longitudinal displacement of group A was more than 3.00 mm, indicating the failure criterion, while the axial stiffness increased gradually. Under 200 N loading, there was no significant difference (P > 0.05) in horizontal displacement, longitudinal displacement, and axial stiffness among 4 groups. When the loading reached 600-1 800 N, significant differences were found in horizontal displacement, longitudinal displacement, and axial stiffness among 4 groups (P < 0.05) except the horizontal displacement between groups C and D (P > 0.05). For acetabular fracture in the quadrilateral area, anterior reconstruction plate-lag screw for internal fixation has highest stability, followed by reconstruction plate combined with trans-plate quadrilateral screws, and they are better than infrapectineal buttress reconstruction plate and infrapectineal buttress locking reconstruction plate.

Formation and resuscitation of viable but nonculturable Salmonella typhi.

PubMed

Zeng, Bin; Zhao, Guozhong; Cao, Xiaohong; Yang, Zhen; Wang, Chunling; Hou, Lihua

2013-01-01

Salmonella typhi is a pathogen that causes the human disease of typhoid fever. The aim of this study was to investigate the viable but nonculturable (VBNC) state of S. typhi. Some samples were stimulated at 4°C or -20°C, while others were induced by different concentrations of CuSO4. Total cell counts remained constant throughout several days by acridine orange direct counting; however, plate counts declined to undetectable levels within 48 hours by plate counting at -20°C. The direct viable counts remained fairly constant at this level by direct viable counting. Carbon and nitrogen materials slowly decreased which indicated that a large population of cells existed in the VBNC state and entered the VBNC state in response to exposure to 0.01 or 0.015 mmol/L CuSO4 for more than 14 or 12 days, respectively. Adding 3% Tween 20 or 1% catalase enabled cells to become culturable again, with resuscitation times of 48 h and 24 h, respectively. The atomic force microscope results showed that cells gradually changed in shape from short rods to coccoids, and decreased in size when they entered the VBNC state. Further animal experiments suggested that resuscitated cells might regain pathogenicity.

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

PubMed Central

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

1978-01-01

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

Cultural, Transcriptomic, and Proteomic Analyses of Water-Stressed Cells of Actinobacterial Strains Isolated from Compost: Ecological Implications in the Fed-Batch Composting Process.

PubMed

Narihiro, Takashi; Kanosue, Yuji; Hiraishi, Akira

2016-06-25

This study was undertaken to examine the effects of water activity (aw) on the viability of actinobacterial isolates from a fed-batch composting (FBC) process by comparing culturability and stainability with 5-cyano-2,3-ditoryl tetrazolium chloride (CTC). The FBC reactor as the source of these bacteria was operated with the daily loading of household biowaste for 70 d. During this period of composting, aw in the reactor decreased linearly with time and reached approximately 0.95 at the end of operation. The plate counts of aerobic chemoorganotrophic bacteria were 3.2-fold higher than CTC-positive (CTC+) counts on average at the fully acclimated stage (after 7 weeks of operation), in which Actinobacteria predominated, as shown by lipoquinone profiling and cultivation methods. When the actinobacterial isolates from the FBC process were grown under aw stress, no significant differences were observed in culturability among the cultures, whereas CTC stainability decreased with reductions in aw levels. A cDNA microarray-based transcriptomic analysis of a representative isolate showed that many of the genes involved in cellular metabolism and genetic information processing were down-regulated by aw stress. This result was fully supported by a proteomic analysis. The results of the present study suggest that, in low aw mature compost, the metabolic activity of the community with Actinobacteria predominating is temporarily reduced to a level that hardly reacts with CTC; however, these bacteria are easily recoverable by exposure to a high aw culture medium. This may be a plausible reason why acclimated FBC reactors in which Actinobacteria predominate yields higher plate counts than CTC+ counts.

MCP performance improvement using alumina thin film

NASA Astrophysics Data System (ADS)

Yang, Yuzhen; Yan, Baojun; Liu, Shulin; Zhao, Tianchi; Yu, Yang; Wen, Kaile; Li, Yumei; Qi, Ming

2017-10-01

The performance improvement using alumina thin film on a dual microchannel plate (MCP) detector for single electron counting was investigated. The alumina thin film was coated on all surfaces of the MCPs by atomic layer deposition method. It was found that the gain, the single electron resolution and the peak-to-valley ratio of the dual MCP detector were significantly enhanced by coating the alumina thin film. The optimum operating conditions of the new dual MCP detector have been studied.

Comparative determination of two probiotics by QCM and OWLS-based immunosensors.

PubMed

Szalontai, Helga; Adányi, Nóra; Kiss, Attila

2014-09-25

The regular consumption of foods containing probiotic bacteria has beneficial physiological effects on the health and the digestion system. There is a need for novel analytical approaches for the determination of these bacteria that are faster than the classical plate counting method. For this purpose, two label-free biosensors were investigated and presented in this paper: Quartz Crystal Microbalance (QCM) and Optical Waveguide Lightmode Spectroscopy (OWLS) based direct immunosensors were developed for real-time direct detection of probiotic bacteria in fermented dairy products. Bifidobacterium bifidum O1356 and Lactobacillus acidophilus O1132 were detected by polyclonal anti-B. bifidum IgG and anti-L. acidophilus IgG immobilized on the sensors' surface. Sulfo-LC-SPDP cross linking agent was used to bind antibodies to the gold surface of the QCM's AT-cut quartz wafer. Concerning OWLS, antibodies were covalently bound to the amino groups of the silanized surface of the waveguide by glutaraldehyde. The dynamic measuring range was found between 1.0E+3 and 5.0E+5CFUmL(-1) in 100 fold diluted fermented milk products by QCM and with OWLS. Considering the current legislation of the probiotic content in probiotic products, the two developed immunosensors can be applied for rapid quantification of L. acidophilus and B. bifidum in fermented milk. These examinations offer effective alternatives to the microbiological plate counting method. Copyright © 2014 Elsevier B.V. All rights reserved.

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

PubMed

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 (<0.45); the majority of R(2) values were <0.25, indicating small relationship between the results from the raw milk 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.

Evaluation of 3 dental unit waterline contamination testing methods

PubMed Central

Porteous, Nuala; Sun, Yuyu; Schoolfield, John

2015-01-01

Previous studies have found inconsistent results from testing methods used to measure heterotrophic plate count (HPC) bacteria in dental unit waterline (DUWL) samples. This study used 63 samples to compare the results obtained from an in-office chairside method and 2 currently used commercial laboratory HPC methods (Standard Methods 9215C and 9215E). The results suggest that the Standard Method 9215E is not suitable for application to DUWL quality monitoring, due to the detection of limited numbers of heterotrophic organisms at the required 35°C incubation temperature. The results also confirm that while the in-office chairside method is useful for DUWL quality monitoring, the Standard Method 9215C provided the most accurate results. PMID:25574718

Simultaneous construction of PCR-DGGE-based predictive models of Listeria monocytogenes and Vibrio parahaemolyticus on cooked shrimps.

PubMed

Liao, C; Peng, Z Y; Li, J B; Cui, X W; Zhang, Z H; Malakar, P K; Zhang, W J; Pan, Y J; Zhao, Y

2015-03-01

The aim of this study was to simultaneously construct PCR-DGGE-based predictive models of Listeria monocytogenes and Vibrio parahaemolyticus on cooked shrimps at 4 and 10°C. Calibration curves were established to correlate peak density of DGGE bands with microbial counts. Microbial counts derived from PCR-DGGE and plate methods were fitted by Baranyi model to obtain molecular and traditional predictive models. For L. monocytogenes, growing at 4 and 10°C, molecular predictive models were constructed. It showed good evaluations of correlation coefficients (R(2) > 0.92), bias factors (Bf ) and accuracy factors (Af ) (1.0 ≤ Bf ≤ Af ≤ 1.1). Moreover, no significant difference was found between molecular and traditional predictive models when analysed on lag phase (λ), maximum growth rate (μmax ) and growth data (P > 0.05). But for V. parahaemolyticus, inactivated at 4 and 10°C, molecular models show significant difference when compared with traditional models. Taken together, these results suggest that PCR-DGGE based on DNA can be used to construct growth models, but it is inappropriate for inactivation models yet. This is the first report of developing PCR-DGGE to simultaneously construct multiple molecular models. It has been known for a long time that microbial predictive models based on traditional plate methods are time-consuming and labour-intensive. Denaturing gradient gel electrophoresis (DGGE) has been widely used as a semiquantitative method to describe complex microbial community. In our study, we developed DGGE to quantify bacterial counts and simultaneously established two molecular predictive models to describe the growth and survival of two bacteria (Listeria monocytogenes and Vibrio parahaemolyticus) at 4 and 10°C. We demonstrated that PCR-DGGE could be used to construct growth models. This work provides a new approach to construct molecular predictive models and thereby facilitates predictive microbiology and QMRA (Quantitative Microbial Risk Assessment). © 2014 The Society for Applied Microbiology.

Comparison of the compact dry TC method with the standard method ISO 21149:2006 for determining aerobic colony counts in cosmetic emulsion.

PubMed

De Vaugelade, S; Aime, M; Farcette, N; Maurel, E; Lacour, T; Thomas, C; Bouchonnet, S; Pirnay, S

2017-02-01

Compact Dry TC, a rapid method kit for determining aerobic colony counts, has been developed by Nissui Pharmaceutical Co. for food application. These plates are pre-sterilized and contain culture medium, a cold-soluble gelling agent and a colour redox indicator for rapid enumeration. In this study, the alternative method is compared with the standard method ISO 21149:2006 - Cosmetic - Microbiology - Enumeration and detection of aerobic mesophilic bacteria, for cosmetic emulsions application. An oil-in-water (o/w) cosmetic emulsion was contaminated with a pool of bacterial strains (Escherichia coli ATCC 8739, Staphylococcus aureus ATCC 6538 and Pseudomonas aeruginosa ATCC 9027). One millilitre of samples was spread on agar as described in ISO 21149. The colonies were enumerated after 3 days of incubation. At the same time, 1.2 mL samples were spread on Compact Dry TC kits. The kit was incubated at 35°C ± 1°C for 48 h, and the colonies were enumerated. Accuracy determination was carried out using six replicates at four levels of concentrations (10, 50, 100 and 250 CFU mL -1 ). The repeatability study was carried out using 12 replicates at four levels of concentrations (10, 50, 100 and 250 CFU mL -1 ). Variations relative to the analyst and to the batch of emulsion have been investigated. The linear correlation coefficients of Compact Dry TC Kit enumeration with standard method ISO 21149:2006 was 0.9999. In comparison study, no apparent differences were noted between the Compact Dry TC kit and the reference method ISO 21149, for the detection level of aerobic microorganisms. Relative accuracy, repeatability and intermediate precision studies were acceptable. In the repeatability study, the Shapiro-Wilk test has confirmed the normally distribution of the twelve assays. No significant variations in Compact Dry TC count results were observed with different analysts and different batches of emulsion. The results showed that the two compared methods 'Compact Dry TC' vs. 'conventional pour plate' performed equally well. Demonstration was achieved that the Compact Dry TC method may constitute a useful alternative tool for rapid enumeration of aerobic mesophilic bacteria in cosmetic emulsions. © 2016 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

Analysis Total Plate Counte (TPC) On Fresh Steak Tuna Applications Edible Coating Caulerpa sp During Stored at Chilling Temperature

NASA Astrophysics Data System (ADS)

Nelce Mailoa, Meigy; Marthina Tapotubun, Alfonsina; Matrutty, Theodora E. A. A.

2017-10-01

A study has been conducted to determine the use of Caulerpa sp. Edible coatings on fresh steaks tuna against the presence of microbes during storage at chilling temperatures. In this research, two applied method of edible coating is used, that is dipping and immersion method, with chilling temperature (5°C), storage time (0, 3, 6, 9 days). Each treatment is compared to control (without an edible coating). The results showed that the application of edible coating Caulerpa sp in fresh steaks tuna with soaking and dipping method showed total bacteria increase during storage, but still fulfill the microbiological quality of fresh fish that is maximum total plate 5.0 × 105 cfu/g. Total microbes in fresh steaks tuna were soaked with immersion methods and stored up to 9 days: 2.3 × 105 cfu/g while total microbial on fresh steaks tuna were dipped by dipping method and stored up to 9 days ie 3.4 × 105 cfu/g. So it can be concluded that application method of edible coating Caulerpa sp on fresh steaks tuna soaked better than the method of dipping.

Airborne Salmonella and Listeria associated with Irish commercial beef, sheep and pig plants.

PubMed

Okraszewska-Lasica, Wioletta; Bolton, D J; Sheridan, J J; McDowell, D A

2014-06-01

Air samples from lairage, hide/fleece pulling or dehairing/scraping, evisceration and chilling areas in commercial beef, sheep and pig plants were examined for Salmonella spp. and Listeria monocytogenes, by impaction or sedimentation onto selective (Brilliant Green Agar, BSA; Listeria Selective Agar, LSA) and non-selective (Plate Count Agar, PCA) media. Both pathogens were frequently detected in all three plants. Improved recoveries were achieved by combining sedimentation, and broth based resuscitation, suggesting cell injury. Salmonella were recovered from all three plants, with the highest counts on BGA in the pig plant. The most common serotypes were S. Typhimurium in the beef/sheep plants and S. Derby in the pig plant. Very low counts of L. monocytogenes (e.g. 2.6CFUm(2)) were detected at hide removal on LSA sedimentation plates in the beef plant. These included serogroup 1/2a-3a and 1/2b-3b-7. Pathogen counts in the three plants were generally very low, suggesting that air is unlikely to be a significant source of carcass or plant surface contamination. Copyright © 2014 Elsevier Ltd. All rights reserved.

Science hub spore data

EPA Pesticide Factsheets

Data set includes UV dose, and Bacillus pumilus spore plate counts in colony forming unitsThis dataset is associated with the following publication:Boczek , L., E. Rhodes , J. Cashdollar, J. Ryu, J. Popovici , J. Hoelle , M. Sivaganesan , S. Hayes , M. Rodgers , and H. Ryu. Applicability of UV resistant Bacillus pumilus endospores as a human adenovirus surrogate for evaluating the effectiveness of virus inactivation in low-pressure UV treatment systems. JOURNAL OF MICROBIOLOGICAL METHODS. Elsevier Science Ltd, New York, NY, USA, 122: 43-49, (2016).

Biofilms associated with poultry processing equipment.

PubMed

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

1996-01-01

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

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

PubMed

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

2014-12-01

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

Counting particles emitted by stratospheric aircraft and measuring size of particles emitted by stratospheric aircraft

NASA Technical Reports Server (NTRS)

Wilson, James Charles

1994-01-01

The ER-2 condensation nuclei counter (CNC) has been modified to reduce the diffusive losses of particles within the instrument. These changes have been successful in improving the counting efficiency of small particles at low pressures. Two techniques for measuring the size distributions of particles with diameters less than 0.17 micrometers have been evaluated. Both of these methods, the differential mobility analyzer (DMA) and the diffusion battery, have fundamental problems that limit their usefulness for stratospheric applications. We cannot recommend either for this application. Newly developed, alternative methods for measuring small particles include inertial separation with a low-loss critical orifice and thin-plate impactor device. This technique is now used to collect particles in the multisample aerosol collector housed in the ER-2 CNC-2, and shows some promise for particle size measurements when coupled with a CNC as a counting device. The modified focused-cavity aerosol spectrometer (FCAS) can determine the size distribution of particles with ambient diameters as small as about 0.07 micrometers. Data from this instrument indicates the presence of a nuclei mode when CNC-2 indicates high concentrations of particles, but cannot resolve important parameters of the distribution.

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

NASA Technical Reports Server (NTRS)

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

1975-01-01

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

Microchannel plate life testing for UV spectroscopy instruments

NASA Astrophysics Data System (ADS)

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

2017-08-01

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

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

PubMed

Berry, Elaine D; Wells, James E

2008-11-01

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

Track counts as indices to abundances of arboreal rodents.

Treesearch

A.B. Carey; J.W. Witt

1991-01-01

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

Detection of E. coli O157:H7 from ground beef using Fourier transform infrared (FT-IR) spectroscopy and chemometrics.

PubMed

Davis, Reeta; Irudayaraj, Joseph; Reuhs, Bradley L; Mauer, Lisa J

2010-08-01

FT-IR spectroscopy methods for detection, differentiation, and quantification of E. coli O157:H7 strains separated from ground beef were developed. Filtration and immunomagnetic separation (IMS) were used to extract live and dead E. coli O157:H7 cells from contaminated ground beef prior to spectral acquisition. Spectra were analyzed using chemometric techniques in OPUS, TQ Analyst, and WinDAS software programs. Standard plate counts were used for development and validation of spectral analyses. The detection limit based on a selectivity value using the OPUS ident test was 10(5) CFU/g for both Filtration-FT-IR and IMS-FT-IR methods. Experiments using ground beef inoculated with fewer cells (10(1) to 10(2) CFU/g) reached the detection limit at 6 h incubation. Partial least squares (PLS) models with cross validation were used to establish relationships between plate counts and FT-IR spectra. Better PLS predictions were obtained for quantifying live E. coli O157:H7 strains (R(2)> or = 0.9955, RMSEE < or = 0.17, RPD > or = 14) and different ratios of live and dead E. coli O157:H7 cells (R(2)= 0.9945, RMSEE = 2.75, RPD = 13.43) from ground beef using Filtration-FT-IR than IMS-FT-IR methods. Discriminant analysis and canonical variate analysis (CVA) of the spectra differentiated various strains of E. coli O157:H7 from an apathogenic control strain. CVA also separated spectra of 100% dead cells separated from ground beef from spectra of 0.5% live cells in the presence of 99.5% dead cells of E. coli O157:H7. These combined separation and FT-IR methods could be useful for rapid detection and differentiation of pathogens in complex foods.

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

DTIC Science & Technology

2014-12-11

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

Analysis of bending wave transmission using beam tracing with advanced statistical energy analysis for periodic box-like structures affected by spatial filtering

NASA Astrophysics Data System (ADS)

Wilson, D.; Hopkins, C.

2015-04-01

For bending wave transmission across periodic box-like arrangements of plates, the effects of spatial filtering can be significant and this needs to be considered in the choice of prediction model. This paper investigates the errors that can occur with Statistical Energy Analysis (SEA) and the potential of using Advanced SEA (ASEA) to improve predictions. The focus is on the low- and mid-frequency range where plates only support local modes with low mode counts and the in situ modal overlap is relatively high. To increase the computational efficiency when using ASEA on large systems, a beam tracing method is introduced which groups together all rays with the same heading into a single beam. Based on a diffuse field on the source plate, numerical experiments are used to determine the angular distribution of incident power on receiver plate edges on linear and cuboid box-like structures. These show that on receiver plates which do not share a boundary with the source plate, the angular distribution on the receiver plate boundaries differs significantly from a diffuse field. SEA and ASEA predictions are assessed through comparison with finite element models. With rain-on-the-roof excitation on the source plate, the results show that compared to SEA, ASEA provides significantly better estimates of the receiver plate energy, but only where there are at least one or two bending modes in each one-third octave band. Whilst ASEA provides better accuracy than SEA, discrepancies still exist which become more apparent when the direct propagation path crosses more than three nominally identical structural junctions.

The influence of dialyzer geometry on blood coagulation and biocompatibility.

PubMed

Lins, L E; Boberg, U; Jacobson, S H; Kjellstrand, C; Ljungberg, B; Skröder, R

1993-11-01

The influence of dialyzer geometry on blood coagulation, heparin requirement and complement activation was studied in fourteen chronic hemodialysis patients. Each patient was dialyzed with two different cuprophan dialyzers, hollow fiber GF 120M and parallel plate Lundia IC5N. Both dialyzers had a wall thickness of 11 microns, surface area of 1.2 m2 and both were sterilized with ethylene oxide. Heparin doses were individually titrated. The mean heparin dose was 6089 +/- 988 U. Platelet count decreased from 218 x 10(9)/l to 193 x 10(9)/l and from 235 x 10(9)/l to 197 x 10(9)/l respectively (hollow fiber/plate dialyzer, ns). The number of leucocytes decreased at 15 min after start of dialysis by 56% and 61% (hollow fiber/plate dialyzer, ns). The heparin requirement, measured as prolongation of whole blood activated coagulation time after identical doses of heparin, were the same in hollow fiber and plate dialysis sessions. The arterial fibrinopeptide A concentrations increased during dialysis from 5.4 to 7.1 nmol/l and 8.5 to 9.6 nmol/l respectively (hollow fiber/plate dialyzer, ns). The residual blood volume in the hollow fiber dialyzers was 1.3 +/- 1.1 ml and in the plate dialyzers 1.5 +/- 0.9 ml (ns). C3a activation, indicated by a marked arterio-venous difference, was observed at 15 min after start of dialysis with hollow fiber as well as plate dialyzers. The arterio-venous difference was less pronounced at the end of dialysis. There were no differences in C3a activation between hollow fiber and plate dialyzers at any timepoint. It is concluded that dialyzer geometry does not significantly influence platelet count, blood coagulation, heparin requirement or complement activation.

Microchannel plate cross-talk mitigation for spatial autocorrelation measurements

NASA Astrophysics Data System (ADS)

Lipka, Michał; Parniak, Michał; Wasilewski, Wojciech

2018-05-01

Microchannel plates (MCP) are the basis for many spatially resolved single-particle detectors such as ICCD or I-sCMOS cameras employing image intensifiers (II), MCPs with delay-line anodes for the detection of cold gas particles or Cherenkov radiation detectors. However, the spatial characterization provided by an MCP is severely limited by cross-talk between its microchannels, rendering MCP and II ill-suited for autocorrelation measurements. Here, we present a cross-talk subtraction method experimentally exemplified for an I-sCMOS based measurement of pseudo-thermal light second-order intensity autocorrelation function at the single-photon level. The method merely requires a dark counts measurement for calibration. A reference cross-correlation measurement certifies the cross-talk subtraction. While remaining universal for MCP applications, the presented cross-talk subtraction, in particular, simplifies quantum optical setups. With the possibility of autocorrelation measurements, the signal needs no longer to be divided into two camera regions for a cross-correlation measurement, reducing the experimental setup complexity and increasing at least twofold the simultaneously employable camera sensor region.

A statistical treatment of bioassay pour fractions

NASA Astrophysics Data System (ADS)

Barengoltz, Jack; Hughes, David

A bioassay is a method for estimating the number of bacterial spores on a spacecraft surface for the purpose of demonstrating compliance with planetary protection (PP) requirements (Ref. 1). The details of the process may be seen in the appropriate PP document (e.g., for NASA, Ref. 2). In general, the surface is mechanically sampled with a damp sterile swab or wipe. The completion of the process is colony formation in a growth medium in a plate (Petri dish); the colonies are counted. Consider a set of samples from randomly selected, known areas of one spacecraft surface, for simplicity. One may calculate the mean and standard deviation of the bioburden density, which is the ratio of counts to area sampled. The standard deviation represents an estimate of the variation from place to place of the true bioburden density commingled with the precision of the individual sample counts. The accuracy of individual sample results depends on the equipment used, the collection method, and the culturing method. One aspect that greatly influences the result is the pour fraction, which is the quantity of fluid added to the plates divided by the total fluid used in extracting spores from the sampling equipment. In an analysis of a single sample’s counts due to the pour fraction, one seeks to answer the question: What is the probability that if a certain number of spores are counted with a known pour fraction, that there are an additional number of spores in the part of the rinse not poured. This is given for specific values by the binomial distribution density, where detection (of culturable spores) is success and the probability of success is the pour fraction. A special summation over the binomial distribution, equivalent to adding for all possible values of the true total number of spores, is performed. This distribution when normalized will almost yield the desired quantity. It is the probability that the additional number of spores does not exceed a certain value. Of course, for a desired value of uncertainty, one must invert the calculation. However, this probability of finding exactly the number of spores in the poured part is correct only in the case where all values of the true number of spores greater than or equal to the adjusted count are equally probable. This is not realistic, of course, but the result can only overestimate the uncertainty. So it is useful. In probability speak, one has the conditional probability given any true total number of spores. Therefore one must multiply it by the probability of each possible true count, before the summation. If the counts for a sample set (of which this is one sample) are available, one may use the calculated variance and the normal probability distribution. In this approach, one assumes a normal distribution and neglects the contribution from spatial variation. The former is a common assumption. The latter can only add to the conservatism (over estimate the number of spores at some level of confidence). A more straightforward approach is to assume a Poisson probability distribution for the measured total sample set counts, and use the product of the number of samples and the mean number of counts per sample as the mean of the Poisson distribution. It is necessary to set the total count to 1 in the Poisson distribution when actual total count is zero. Finally, even when the planetary protection requirements for spore burden refer only to the mean values, they require an adjustment for pour fraction and method efficiency (a PP specification based on independent data). The adjusted mean values are a 50/50 proposition (e.g., the probability of the true total counts in the sample set exceeding the estimate is 0.50). However, this is highly unconservative when the total counts are zero. No adjustment to the mean values occurs for either pour fraction or efficiency. The recommended approach is once again to set the total counts to 1, but now applied to the mean values. Then one may apply the corrections to the revised counts. It can be shown by the methods developed in this work that this change is usually conservative enough to increase the level of confidence in the estimate to 0.5. 1. NASA. (2005) Planetary protection provisions for robotic extraterrestrial missions. NPR 8020.12C, April 2005, National Aeronautics and Space Administration, Washington, DC. 2. NASA. (2010) Handbook for the Microbiological Examination of Space Hardware, NASA-HDBK-6022, National Aeronautics and Space Administration, Washington, DC.

Antimicrobial and anti-adherence activity of various combinations of coffee-chicory solutions on Streptococcus mutans: An in-vitro study

PubMed Central

Sharma, Rama; Reddy, Vamsi Krishna L; Prashant, GM; Ojha, Vivek; Kumar, Naveen PG

2014-01-01

Context: Several studies have demonstrated the activity of natural plants on the dental biofilm and caries development. But few studies on the antimicrobial activity of coffee-based solutions were found in the literature. Further there was no study available to check the antimicrobial effect of coffee solutions with different percentages of chicory in it. Aims: To evaluate the antimicrobial activity of different combinations of coffee-chicory solutions and their anti-adherence effect on Streptococcus mutans to glass surface. Materials and Methods: Test solutions were prepared. For antimicrobial activity testing, tubes containing test solution and culture medium were inoculated with a suspension of S. mutans followed by plating on Brain Heart Infusion (BHI) agar. S. mutans adherence to glass in presence of the different test solutions was also tested. The number of adhered bacteria (CFU/mL) was determined by plating method. Statistical Analysis: Statistical significance was measured using one way ANOVA followed by Tukey's post hoc test. P value < 0.05 was considered statistically significant. Results: Pure chicory had shown significantly less bacterial count compared to all other groups. Groups IV and V had shown significant reduction in bacterial counts over the period of 4 hrs. Regarding anti-adherence effect, group I-IV had shown significantly less adherence of bacteria to glass surface. Conclusions: Chicory exerted antibacterial effect against S. mutans while coffee reduced significantly the adherence of S. mutans to the glass surface. PMID:25328299

Moxa-stick suffumigation for disinfecting air in hematology and hematopoietic stem cell transplantation wards with class 100 laminar flow.

PubMed

He, Jing-song; Yang, Qing; Huang, Wei-jia; Hu, Xiao-rong

2014-04-01

To evaluate the effect of moxa-stick suffumigation in the hematology and hematopoietic stem cell transplantation (HSCT) wards with luminar flow. The plate exposure method was used to measure the effect of air-disinfection of moxa-stick suffumigation in hematology and HSCT wards. The yearly average qualified rates of air sampling in HSCT wards were evaluated from 2007 to 2010. To further investigate the disinfecting effect of moxa-stick suffumigation, the colony counts of common pathogens (including Staphylcoccus aureus and Pseudomonas aeruginosa) before and after moxa-stick suffumigation were compared. The mean air quality rates of the HSCT wards with class 100 laminar flow were all above 90.0% (91.2%-96.2%) from 2007 to 2010. Moxa-stick suffumigation effectively decreased the presence of bacteria in the hematology ward's air (P<0.01). The most notable effect was the drastic reduction in the colony counts of Staphylococcus aureus and Pseudomonas aeruginosa on the blood plates exposed to air treated with moxa-stick suffumigation (77.1±52.9 cfu/m(2) vs 196.1±87.5 cfu/m(2), P<0.01; and 100.2±35.3 cfu/m(2) vs 371.5±35.3 cfu/m(2), P<0.01). Moxa-stick suffumigation proved to be a reliable and effective airdisinfection method for hematology and HSCT wards, and hence, it should be employed extensively.

High throughput screening of active pharmaceutical ingredients by UPLC.

PubMed

Al-Sayah, Mohammad A; Rizos, Panagiota; Antonucci, Vincent; Wu, Naijun

2008-07-01

Ultra performance LC (UPLC) was evaluated as an efficient screening approach to facilitate method development for drug candidates. Three stationary phases were screened: C-18, phenyl, and Shield RP 18 with column dimensions of 150 mm x 2.1 mm, 1.7 microm, which should theoretically generate 35,000 plates or 175% of the typical column plate count of a conventional 250 mm x 4.6 mm, 5 microm particle column. Thirteen different active pharmaceutical ingredients (APIs) were screened using this column set with a standardized mobile-phase gradient. The UPLC method selectivity results were compared to those obtained for these compounds via methods developed through laborious trial and error screening experiments using numerous conventional HPLC mobile and stationary phases. Peak capacity was compared for columns packed with 5 microm particles and columns packed with 1.7 microm particles. The impurities screened by UPLC were confirmed by LC/MS. The results demonstrate that simple, high efficiency UPLC gradients are a feasible and productive alternative to more conventional multiparametric chromatographic screening approaches for many compounds in the early stages of drug development.

Rapid in situ assessment of physiological activities in bacterial biofilms using fluorescent probes

NASA Technical Reports Server (NTRS)

Yu, F. P.; McFeters, G. A.

1994-01-01

Two rapid in situ enumeration methods using fluorescent probes were used to assess the physiological activities of Klebsiella pneumoniae biofilms on stainless steel. Fluorescent dyes, 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and rhodamine 123 (Rh 123), were chosen to perform this study. CTC is a soluble redox indicator which can be reduced by respiring bacteria to fluorescent CTC-formazan crystals. Rh 123 is incorporated into bacteria with respect to cellular proton motive force. The intracellular accumulation of these fluorescent dyes can be determined using epifluorescence microscopy. The results obtained with these two fluorescent probes in situ were compared to the plate count (PC) and in situ direct viable count (DVC) methods. Viable cell densities within biofilms determined by the three in situ methods were comparable and always showed approximately 2-fold higher values than those obtained with the PC method. As an additional advantage, the results were observed after 2 h, which was shorter than the 4 h incubation time required for the DVC method and 24 h for colony formation. The results indicate that staining with CTC and Rh 123 provides rapid information regarding cell numbers and physiological activities of bacteria within biofilms.

Embedded and conventional ultrasonic sensors for monitoring acoustic emission during thermal fatigue

NASA Astrophysics Data System (ADS)

Trujillo, Blaine; Zagrai, Andrei

2016-04-01

Acoustic emission is widely used for monitoring pressure vessels, pipes, critical infrastructure, as well as land, sea and air vehicles. It is one of dominant approaches to explore material degradation under fatigue and events leading to material fracture. Addressing a recent interest in structural health monitoring of space vehicles, a need has emerged to evaluate material deterioration due to thermal fatigue during spacecraft atmospheric reentry. Thermal fatigue experiments were conducted, in which aluminum plates were subjected to localized heating and acoustic emission was monitoring by embedded and conventional acoustic emission sensors positioned at various distances from a heat source. At the same time, surface temperature of aluminum plates was monitored using an IR camera. Acoustic emission counts collected by embedded sensors were compared to counts measured with conventional acoustic emission sensors. Both types of sensors show noticeable increase of acoustic emission activity as localized heating source was applied to aluminum plates. Experimental data demonstrate correlation between temperature increase on the surface of the plates and increase in measured acoustic emission activity. It is concluded that under particular conditions, embedded piezoelectric wafer active sensors can be used for acoustic emission monitoring of thermally-induced structural degradation.

Virological and bacteriological quality of drinking water in Ethiopia

NASA Astrophysics Data System (ADS)

Bedada, Tesfaye Legesse; Mezemir, Walelign Dessie; Dera, Firehiwot Abera; Sima, Waktole Gobena; Gebre, Samson Girma; Edicho, Redwan Muzeyin; Biegna, Almaz Gonfa; Teklu, Dejenie Shiferaw; Tullu, Kassu Desta

2018-05-01

Since unsafe water is responsible for many illness, deaths, and economic failure, water quality monitoring is essential. A cross-sectional study was conducted on 218 drinking waters samples collected between February and June 2016 to assess water quality using phages by the help of CB390 E. coli host, plaque assay; multiple tube fermentation for coliforms and pour plate for heterotrophic bacteria at Ethiopian Public Health Institute. Heterotrophic plate count greater than 100 cfu/ml was noted in 41 samples and detections of total and thermotolerant coliforms and E. coli in 38, 24, and 10 samples, respectively, and no phages detection in chlorinated waters. While heterotrophic plate count greater than 100 cfu/ml was observed in 100 samples and detections of total and thermotolerant coliforms, E. coli, and phages in 75, 60, 42, and 5 samples, respectively, for untreated waters. The majority of the waters contained indicators above standard limits. This indicates that the sources are contaminated and they are potential threats for health. Hence, regular water monitoring should be a priority agenda.

A direct viable count method for the enumeration of attached bacteria and assessment of biofilm disinfection

NASA Technical Reports Server (NTRS)

Yu, F. P.; Pyle, B. H.; McFeters, G. A.

1993-01-01

This report describes the adaptation of an in situ direct viable count (in situ DVC) method in biofilm disinfection studies. The results obtained with this technique were compared to two other enumeration methods, the plate count (PC) and conventional direct viable count (c-DVC). An environmental isolate (Klebsiella pneumoniae Kp1) was used to form biofilms on stainless steel coupons in a stirred batch reactor. The in situ DVC method was applied to directly assess the viability of bacteria in biofilms without disturbing the integrity of the interfacial community. As additional advantages, the results were observed after 4 h instead of the 24 h incubation time required for colony formation and total cell numbers that remained on the substratum were enumerated. Chlorine and monochloramine were used to determine the susceptibilities of attached and planktonic bacteria to disinfection treatment using this novel analytical approach. The planktonic cells in the reactor showed no significant change in susceptibility to disinfectants during the period of biofilm formation. In addition, the attached cells did not reveal any more resistance to disinfection than planktonic cells. The disinfection studies of young biofilms indicated that 0.25 mg/l free chlorine (at pH 7.2) and 1 mg/l monochloramine (at pH 9.0) have comparable disinfection efficiencies at 25 degrees C. Although being a weaker disinfectant, monochloramine was more effective in removing attached bacteria from the substratum than free chlorine. The in situ DVC method always showed at least one log higher viable cell densities than the PC method, suggesting that the in situ DVC method is more efficient in the enumeration of biofilm bacteria. The results also indicated that the in situ DVC method can provide more accurate information regarding the cell numbers and viability of bacteria within biofilms following disinfection.

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

NASA Technical Reports Server (NTRS)

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

1999-01-01

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

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

PubMed

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

2015-04-01

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

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

PubMed

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.

Limitations in the use of ozone to disinfect maple sap.

PubMed

Labbe, R G; Kinsley, M; Wu, J

2001-01-01

The sap of the maple sugar tree (Acer saccharum) contains 2 to 3% sucrose and is traditionally collected early in the year and concentrated by boiling to produce maple syrup. High levels of microorganisms in the sap occur during holding, leading to a darker syrup with lower economic value. We investigated the use of dissolved ozone as a method to reduce the microbial population in sap. After 40 min of ozone treatment, concentrations of up to 0.30 mg/liter were achieved but were ineffective in reducing the aerobic plate count. Three predominant colonies on nutrient agar were selected for isolation and identification from sap. These included one mucoid and one nonmucoid yeast, both identified as Candida, and Pseudomonas fluorescens. When suspended in buffer, each was readily inactivated by ozone. Addition of 3% sucrose to the buffer markedly reduced the effectiveness of ozone. With the use of an ozone generator with a larger ozone output, saturating ozone concentrations (1 mg/liter) were achieved within 5 min but were accompanied by only a 1-log reduction in aerobic plate count of maple sap. After 40 min of ozone treatment, a less than 3-log reduction occurred. The results indicate that, because of the presence of sucrose, ozone may be of limited use in reducing the microbial population in sap.

Bacteriological quality of the wastewater used for irrigation at the vegetable farms in Korle-bu Teaching Hospital, Accra Metropolis, Ghana.

PubMed

Pesewu, George A; Bentum, Daniel; Olu-Taiwo, Michael A; Glover, Kathreen K; Yirenya-Tawiah, Dzidzo R

2017-01-01

Many developing countries, including Ghana, are water stressed. As such, farmers, particularly those in urban areas, have adopted the use of wastewater for irrigation. This study evaluated the bacteriological water quality of the wastewater used for irrigation in the vegetable farms at Korle-Bu Teaching Hospital (KBTH), Accra Metropolis, Ghana. In all, 40 wastewater samples were collected and analysed bacteriologically using the total aerobic plate count method. The isolated bacteria were identified biochemically using Bergey's manual for determinative bacteriology. Mean total bacterial colony count values in the range of 2.75-4.44 × 10 5 CFU/100 mL were isolated which far exceeds values of 1 × 10 3 /100 mL recommended by the World Health Organization (WHO) for unrestricted irrigation of crops likely to be eaten raw. Enterobacter cloacae (51.4%), Klebsiella sp. (24.1%), Pseudomonas aeruginosa (11.3%), Salmonella typhi (10.6%), Escherichia coli (2.2%) and Proteus sp. (0.4%) were the predominant bacteria isolated. Growers should use treated wastewater for farming while processors and consumers should minimize contamination risks of produce from the vegetable farms/garden to the plate. © The Author(s) 2016.

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

NASA Technical Reports Server (NTRS)

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

2003-01-01

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

A method for detecting fungal contaminants in wall cavities.

PubMed

Spurgeon, Joe C

2003-01-01

This article describes a practical method for detecting the presence of both fungal spores and culturable fungi in wall cavities. Culturable fungi were collected in 25 mm cassettes containing 0.8 microm mixed cellulose ester filters using aggressive sampling conditions. Both culturable fungi and fungal spores were collected in modified slotted-disk cassettes. The sample volume was 4 L. The filters were examined microscopically and dilution plated onto multiple culture media. Collecting airborne samples in filter cassettes was an effective method for assessing wall cavities for fungal contaminants, especially because this method allowed the sample to be analyzed by both microscopy and culture media. Assessment criteria were developed that allowed the sample results to be used to classify wall cavities as either uncontaminated or contaminated. As a criterion, wall cavities with concentrations of culturable fungi below the limit of detection (LOD) were classified as uncontaminated, whereas those cavities with detectable concentrations of culturable fungi were classified as contaminated. A total of 150 wall cavities was sampled as part of a field project. The concentrations of culturable fungi were below the LOD in 34% of the samples, whereas Aspergillus and/or Penicillium were the only fungal genera detected in 69% of the samples in which culturable fungi were detected. Spore counting resulted in the detection of Stachybotrys-like spores in 25% of the samples that were analyzed, whereas Stachybotrys chartarum colonies were only detected on 2% of malt extract agar plates and on 6% of corn meal agar plates.

Principle component analysis (PCA) for investigation of relationship between population dynamics of microbial pathogenesis, chemical and sensory characteristics in beef slices containing Tarragon essential oil.

PubMed

Alizadeh Behbahani, Behrooz; Tabatabaei Yazdi, Farideh; Shahidi, Fakhri; Mortazavi, Seyed Ali; Mohebbi, Mohebbat

2017-04-01

Principle component analysis (PCA) was employed to examine the effect of the exerted treatments on the beef shelf life as well as discovering the correlations between the studied responses. Considering the variability of the dimensions of the responses, correlation coefficients were applied to form the matrix and extract the eigenvalue. Antimicrobial effect was evaluated on 10 pathogenic microorganisms through the methods of hole-plate diffusion method, disk diffusion method, pour plate method, minimum inhibitory concentration and minimum bactericidal/fungicidal concentration. Antioxidant potential and total phenolic content were examined through the method of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Folin-Ciocalteu method, respectively. The components were identified through gas chromatography and gas chromatography/mass spectrometry. Barhang seed mucilage (BSM) based edible coating containing 0, 0.5, 1 and 1.5% (w/w) Tarragon (T) essential oil mix were applied on beef slices to control the growth of pathogenic microorganisms. Microbiological (total viable count, psychrotrophic count, Escherichia coli, Staphylococcus aureus and fungi), chemical (thiobarbituric acid, peroxide value and pH) and sensory characteristics (odor, color and overall acceptability) analysis measurements were made during the storage periodically. PCA was employed to examine the effect of the exerted treatments on the beef shelf life as well as discovering the correlations between the studied responses. Considering the variability of the dimensions of the responses, correlation coefficients were applied to form the matrix and extract the eigenvalue. The PCA showed that the properties of the uncoated meat samples on the 9th, 12th, 15th and 18th days of storage are continuously changing independent of the exerted treatments on the other samples. This reveals the effect of the exerted treatments on the samples. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

PubMed

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

2014-12-01

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

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

NASA Astrophysics Data System (ADS)

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 comparative evaluation of Oratest with the microbiological method of assessing caries activity in children

PubMed Central

Sundaram, Meenakshi; Nayak, Ullal Anand; Ramalingam, Krishnakumar; Reddy, Venugopal; Rao, Arun Prasad; Mathian, Mahesh

2013-01-01

Aims: The aim of this study is to find out whether Oratest can be used as a diagnostic tool in assessing the caries activity by evaluating its relationship to the existing caries status and the salivary streptococcus mutans level. Materials and Methods: The study sample consists of 90 students divided into two groups. Group I (test group) and Group II (control group) consisting of 30 children for control group and 60 children for test group. The sampling of unstimulated saliva for the estimation of streptococcus mutans was done as per the method suggested by Kohler and Bratthall. The plates were then incubated. Rough surface colonies were identified as streptococcus mutans on a pre-determined area of the tip (approximately 1.5 cm2) were counted for each side of spatula pressed against mitis salivarius bacitracin agar using digital colony counter. The results were expressed in colony forming units (CFU). Oratest was carried out in the same patients after the collection of salivary sample for the microbiological method to evaluate the relationship between the two tests. Statistical Analysis Used: The tests used were ANOVA, Pearson Chi-square test, Pearson′s correlation analysis, Mann-Whitney U test and Student′s independent t-test. Results: In the control group and test group, when the streptococcus mutans count (CFU) and Oratest time (minutes) were correlated using Pearson′s correlation analysis, the streptococcus mutans counts was found to be in a statistically significant negative linear relationship with the Oratest time. When the caries status of the children, participated in the test group were correlated with mutans count (CFU) and Oratest time, caries status were found to be in a statistically significant positive linear relationship with streptococcus mutans count and in a significant negative linear relationship with Oratest time. Conclusions: The test proved to be a simple, inexpensive and rapid technique for assessing caries activity since a significant relationship exists clinically with caries status and microbiologically with the streptococcus mutans count of the individual. PMID:23946577

Is there an association between airborne and surface microbes in the critical care environment?

PubMed

Smith, J; Adams, C E; King, M F; Noakes, C J; Robertson, C; Dancer, S J

2018-04-09

There are few data and no accepted standards for air quality in the intensive care unit (ICU). Any relationship between airborne pathogens and hospital-acquired infection (HAI) risk in the ICU remains unknown. First, to correlate environmental contamination of air and surfaces in the ICU; second, to examine any association between environmental contamination and ICU-acquired staphylococcal infection. Patients, air, and surfaces were screened on 10 sampling days in a mechanically ventilated 10-bed ICU for a 10-month period. Near-patient hand-touch sites (N = 500) and air (N = 80) were screened for total colony count and Staphylococcus aureus. Air counts were compared with surface counts according to proposed standards for air and surface bioburden. Patients were monitored for ICU-acquired staphylococcal infection throughout. Overall, 235 of 500 (47%) surfaces failed the standard for aerobic counts (≤2.5 cfu/cm 2 ). Half of passive air samples (20/40: 50%) failed the 'index of microbial air' contamination (2 cfu/9 cm plate/h), and 15/40 (37.5%) active air samples failed the clean air standard (<10 cfu/m 3 ). Settle plate data were closer to the pass/fail proportion from surfaces and provided the best agreement between air parameters and surfaces when evaluating surface benchmark values of 0-20 cfu/cm 2 . The surface standard most likely to reflect hygiene pass/fail results compared with air was 5 cfu/cm 2 . Rates of ICU-acquired staphylococcal infection were associated with surface counts per bed during 72h encompassing sampling days (P = 0.012). Passive air sampling provides quantitative data analogous to that obtained from surfaces. Settle plates could serve as a proxy for routine environmental screening to determine the infection risk in ICU. Copyright © 2018 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

Differential enumeration of subpopulations in concentrated frozen and lyophilized cultures of Lactobacillus delbrueckii ssp. bulgaricus.

PubMed

Shao, Yuyu; Wang, Zhaoxia; Bao, Qiuhua; Zhang, Heping

2017-11-01

Differential enumeration of subpopulations in concentrated frozen and lyophilized cultures of Lactobacillus delbrueckii ssp. bulgaricus ND02 derived from 2 propagation procedures was determined. The subpopulations consisted of 3 categories (physiological states): viable cells capable of forming colonies on agar plates (VC+), viable cells incapable of forming colonies on agar plates (VC-), widely referred to as viable but nonculturable (VBNC) cells, and nonviable or dead cells (NVC). Counts of VC+ were recorded using a conventional plate count procedure. A fluorescent vital staining procedure that discriminates between viable (VC+ and VC-) and NVC cells was used to determine the number of viable and nonviable cells. Both propagation procedures had 2 variables: in procedure (P)1, the propagation medium was rich in yeast extract (4.0%) and the pH was maintained at 5.7; in P2, the medium was devoid of yeast extract and the pH was maintained at 5.1. The results showed that post-propagation operations-concentration of cells by centrifugation and subsequent freezing or lyophilization of cell concentrate-induced different degrees of transience from VC+ to VC- states in cells derived from P1 and P2. Compared with cells derived from P2, cells from P1 were more labile to stress associated with centrifugation, freezing, and lyophilization, as revealed by differential counting. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

PubMed

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

2013-12-01

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

In vitro study on the effect of doxycycline on the microbial activity of soil determined by redox-potential measuring system.

PubMed

Szakmár, Katalin; Reichart, Olivér; Szatmári, István; Erdősi, Orsolya; Szili, Zsuzsanna; László, Noémi; Székely Körmöczy, Péter; Laczay, Péter

2014-09-01

The potential effect of doxycycline on the microbial activity was investigated in three types of soil. Soil samples were spiked with doxycycline, incubated at 25°C and tested at 0, 2, 4 and 6 days after treatment. The microbiological activity of the soil was characterized by the viable count determined by plate pouring and by the time necessary to reach a defined rate of the redox-potential decrease termed as time to detection (TTD).The viable count of the samples was not changed during the storage. The TTD values, however exhibited a significant increase in the 0.2-1.6 mg/kg doxycycline concentration range compared to the untreated samples indicating concentration-dependent inhibitory effect on microbial activity. The potency of the effect was different in the 3 soil types. To describe the combined effect of the doxycycline concentration and time on the biological activity of one type of soil a mathematical model was constructed and applied.The change of microbial metabolic rate could be measured also without (detectable) change of microbial count when the traditional microbiological methods are not applicable. The applied new redox potential measurement-based method is a simple and useful procedure for the examination of microbial activity of soil and its potential inhibition by antibiotics.

Distribution and prevalence of airborne microorganisms in three commercial poultry processing plants.

PubMed

Whyte, P; Collins, J D; McGill, K; Monahan, C; O'Mahony, H

2001-03-01

Airborne microbial contaminants and indicator organisms were monitored within three poultry processing plants (plants A, B, and C). In total, 15 cubic feet (c.f.) of air was sampled per location during 15 visits to each plant and quantitatively analyzed for total mesophilic and psychrophilic aerobic counts, thermophilic campylobacters, Escherichia coli, and Enterobacteriaceae. The prevalence of Salmonella spp. in air samples was also evaluated. Significant reductions in total aerobic counts were observed between defeathering and evisceration areas of the three plants (P < 0.05). Mesophilic plate counts were highest in the defeathering areas of all plants compared to equivalent psychrophilic plate counts. Enterobacteriaceae counts were highest in the defeathering areas of all three plants with counts of log10 1.63, 1.53, and 1.18 CFU/15 c.f. recovered in plants A, B, and C, respectively. E. coli enumerated from air samples in the defeathering areas exhibited a similar trend to those obtained for Enterobacteriaceae with log10 1.67, 1.58, and 1.18 CFU for plants A, B, and C, respectively. Thermophilic campylobacters were most frequently isolated from samples in the defeathering areas followed by the evisceration areas. The highest mean counts of the organism were observed in plant A at 21 CFU/15 c.f. sample with plants B and C at 9 and 8 CFU/sample, respectively. With the exception of low levels of Enterobacteriaceae recovered from samples in the on-line air chill in plant A, E. coli, Enterobacteriaceae, or Campylobacter spp. were not isolated from samples in postevisceration sites in any of the plants examined. Salmonella spp. were not recovered from any samples during the course of the investigation.

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

PubMed

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

2016-06-01

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

The degree of bacterial contamination while performing spine surgery.

PubMed

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

2013-03-01

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

Impact of oral Lactobacillus acidophilus gavage on rooster seminal and cloacal Lactobacilli concentrations.

PubMed

Kiess, A S; Hirai, J H; Triplett, M D; Parker, H M; McDaniel, C D

2016-08-01

The use of antibiotics in poultry is being heavily scrutinized, therefore alternatives such as probiotics are being investigated. Lactobacilli spp. are a commonly used bacteria in formulating probiotics, and the addition of Lactobacilli to broiler diets has demonstrated increased growth rates, stimulated immune systems, and reduced pathogen loads in the gastro-intestinal tract ( GI: ) tract. However, previous research has shown that when rooster semen is directly exposed to Lactobacillus acidophilus (L. acidophilus) sperm quality is reduced. Therefore, the objective of the current study was to determine if oral administration of L. acidophilus increases the concentration of Lactobacilli in semen as well as the cloaca. A total of 30 roosters were used: 15 roosters were gavaged with 1X PBS (Control) and 15 roosters were gavaged with 10(7) cfu/mL of L. acidophilus (Treated). All roosters were gavaged for 14 consecutive days. Semen was collected on a 3 d interval, and cloacal swabs were collected on a 2 d interval, beginning on the first day prior to oral administration. Semen and cloacal swabs were serial diluted, and 100 μL of each dilution was then plated on Man, Rogosa, Sharpe ( MRS: ) agar plates. All plates were incubated for 48 h at 37°C under anaerobic conditions and counted. All Lactobacilli counts were first log transformed, then log transformed (day 0) pre-counts were subtracted from the log transformed day counts providing log differences for the analysis. Seminal Lactobacilli counts were not altered by treatments. However, the main effect of treatment (P = 0.026) for cloacal counts indicated that roosters gavaged with Lactobacilli yielded higher counts than the controls. Additionally, cloaca samples also demonstrated a treatment by day interaction trend (P = 0.082), where Lactobacilli was higher in the L. acidophilus gavaged roosters than the controls only on days 3, 5, 13, and 15. In conclusion, the addition of L. acidophilus to the male breeder diet over extended periods may increase concentrations of Lactobacilli in the cloaca even higher than the concentrations observed in this study. If Lactobacilli reaches high enough concentrations in the cloaca, then sperm quality may be impacted which could lead to poor fertility within the breeder flock. © 2016 Poultry Science Association Inc.

Assessment of the environmental microbiological cross contamination following hand drying with paper hand towels or an air blade dryer.

PubMed

Margas, E; Maguire, E; Berland, C R; Welander, F; Holah, J T

2013-08-01

This study compared the potential for cross contamination of the surrounding environment resulting from two different hand-drying methods: paper towels and the use of an air blade dryer. One hundred volunteers for each method washed their hands and dried them using one of the two methods. Bacterial contamination of the surrounding environment was measured using settle plates placed on the floor in a grid pattern, air sampling and surface swabs. Both drying methods produced ballistic droplets in the immediate vicinity of the hand-drying process. The air blade dryer produced a larger number of droplets which were dispersed over a larger area. Settle plates showed increased microbial contamination in the grid squares which were affected by ballistic droplets. Using the settle plates counts, it was estimated that approx. 1.7 × 10(5) cfu more micro-organisms were left on the laboratory floor (total area approx. 17.15 m(2)) after 100 volunteers used an air blade dryer compared to when paper towels were used. The two drying methods led to different patterns of ballistic droplets and levels of microbial contamination under heavy use conditions. Whilst the increase in microbial levels in the environment is not significant if only nonpathogenic micro-organisms are spread, it may increase the risk of pathogen contamination of the environment when pathogens are occasionally present on people's hands. The study suggests that the risk of cross contamination from the washroom users to the environment and subsequent users should be considered when choosing a hand-drying method. The data could potentially give guidance following the selection of drying methods on implementing measures to minimise the risk of cross contamination. © 2013 The Society for Applied Microbiology.

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

PubMed

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.

Reborn quadrant anode image sensor

NASA Astrophysics Data System (ADS)

Prokazov, Yury; Turbin, Evgeny; Vitali, Marco; Herzog, Andreas; Michaelis, Bernd; Zuschratter, Werner; Kemnitz, Klaus

2009-06-01

We describe a position sensitive photon counting microchannel plate based detector with an improved quadrant anode (QA) readout system. The technique relies on a combination of the four planar elements pattern and an additional fifth electrode. The charge cloud induced by single particle detection is split between the electrodes. The measured charge values uniquely define the position of the initial event. QA has been first published in 1976 by Lampton and Malina. This anode configuration was undeservedly forgotten and its potential has been hardly underestimated. The presented approach extends the operating spatial range to the whole sensitive area of the microchannel plate surface and demonstrates good linearity over the field of view. Therefore, the novel image sensor results in spatial resolution better then 50 μm and count rates up to one million events per second.

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

PubMed

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

2015-04-01

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

Influence of Temperature and Predation on Survival of Salmonella enterica Serovar Typhimurium and Expression of invA in Soil and Manure-Amended Soil▿

PubMed Central

García, R.; Bælum, J.; Fredslund, L.; Santorum, P.; Jacobsen, C. S.

2010-01-01

The effects of three temperatures (5, 15, and 25°C) on the survival of Salmonella enterica serovar Typhimurium in topsoil were investigated in small microcosms by three different techniques: plate counting, invA gene quantification, and invA mRNA quantification. Differences in survival were related to the effect of protozoan predation. Tetracycline-resistant Salmonella serovar Typhimurium was inoculated into soil and manure-amended soil at 1.5 × 108 cells g soil−1. Population densities were determined by plate counting and by molecular methods and monitored for 42 days. Simultaneous extraction of RNA and DNA, followed by quantitative PCR, was used to investigate invA gene levels and expression. Analysis by these three techniques showed that Salmonella serovar Typhimurium survived better at 5°C. Comparing DNA and CFU levels, significantly higher values were determined by DNA-based techniques. invA mRNA levels showed a fast decrease in activity, with no detectable mRNA after an incubation period of less than 4 days in any of the soil scenarios. A negative correlation was found between Salmonella serovar Typhimurium CFU levels and protozoan most probable numbers, and we propose the role of the predator-prey interaction as a factor to explain the die-off of the introduced strain by both culture- and DNA quantification-based methods. The results indicate that temperature, manure, and protozoan predation are important factors influencing the survival of Salmonella serovar Typhimurium in soil. PMID:20562283

Single electron counting using a dual MCP assembly

NASA Astrophysics Data System (ADS)

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

2016-09-01

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

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

USGS Publications Warehouse

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

2003-01-01

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

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

PubMed

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.

Use of mixed cultures of biocontrol agents to control sheep nematodes.

PubMed

Baloyi, M A; Laing, M D; Yobo, K S

2012-03-23

Biological control is a promising non-chemical approach for the control of gastrointestinal nematodes of sheep. Use of combinations of biocontrol agents have been reported to be an effective method to increase the efficacy of biological control effects. In this study, combinations of either two Bacillus thuringiensis (Bt) or Clonostachys rosea (C. rosea) isolates and Bt+C. rosea isolates were evaluated in vitro in microtitre plates for their biocontrol activity on sheep nematodes. The Baermann technique was used to extract the surviving L3 larval stages of intestinal nematodes and counted under a dissecting microscope to determine the larval counts. Results indicate that there was a significant reduction of nematode counts due to combination of biocontrol agents (P<0.001). Combinations of Bt isolates reduced nematodes counts by 72.8%, 64% and 29.8%. The results revealed a control level of 57% when C. rosea isolates P3+P8 were combined. Combination of Bt and C. rosea isolates B10+P8 caused the greatest mortality of 76.7%. Most combinations were antagonistic, with only a few combinations showing an additive effect. None were synergistic. The isolate combinations were more effective than when isolates were used alone. Copyright © 2011 Elsevier B.V. All rights reserved.

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

PubMed

Ingham, S C; Hu, Y; Ané, C

2011-08-01

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

Bactericidal effects of negative air ions on airborne and surface Salmonella enteritidis from an artificially generated aerosol.

PubMed

Seo, K H; Mitchell, B W; Holt, P S; Gast, R K

2001-01-01

The bactericidal effect of high levels of negative ions was studied using a custom-built electrostatic space charge device. To investigate whether the ion-enriched air exerted a bactericidal effect, an aerosol containing Salmonella Enteritidis (SE) was pumped into a sealed plastic chamber. Plates of XLT4 agar were attached to the walls, top, and bottom of the chamber and exposed to the aerosol for 3 h with and without the ionizer treatment. The plates were then removed from the chamber, incubated at 37 degrees C for 24 h, and colonies were counted. An average of greater than 10(3) CFU/plate were observed on plates exposed to the aerosol without the ionizer treatment (control) compared with an average of less than 53 CFU/plate on the ionizer-treated plates. In another series of experiments, the SE aerosol was pumped for 3 h into an empty chamber containing only the ionizer and allowed to collect on the internal surfaces. The inside surfaces of the chamber were then rinsed with 100 ml phosphate-buffered saline that was then plated onto XLT4 plates. While the rinse from the control chamber contained colony counts greater than 400 CFU/ml of wash, no colonies were found in the rinse from the ionizer-treatment chamber. These results indicate that high levels of negative air ions can have a significant impact on the airborne microbial load, and that most of this effect is through direct killing of the organisms. This technology, which also causes significant reduction in airborne dust, has already been successfully applied for poultry hatching cabinets and caged layer rooms. Other potential applications include any enclosed space such as food processing areas, medical institutions, the workplace, and the home, where reduction of airborne and surface pathogens is desired.

The effect of different gas permeability of packaging on physicochemical and microbiological parameters of pork loin storage under high O2 modified atmosphere packaging conditions.

PubMed

Marcinkowska-Lesiak, Monika; Poławska, Ewa; Wierzbicka, Agnieszka

2017-03-01

The aim of this study was to determine the influence of different packaging materials on meat quality during cold storage. Therefore pork loins (m. longissimus thoracis et lumborum) obtained from crossbred pigs (Polish Landrance x Duroc, n = 6) were stored at 2 ℃ in modified atmosphere packs (80% O 2 , 20% CO 2 ) in four types of trays, which differ in gas permeability. Physicochemical (headspace gas composition, pH, colour, drip loss, cooking loss, shear force, the basic composition and fatty acid profile) and microbiological ( Salmonella spp., Escherichia coli, Enterobacteriaceae, total aerobic plates count, total psychrotrophic bacteria count, the number of lactic acid bacteria, Pseudomonas spp., the general amount of yeast and mold) parameters were monitored for up to 12 days. At the end of the storage period no differences in most physicochemical properties of pork loin due to type of packaging were found, however trays with high gas permeability had the greatest impact on total aerobic plates count and Pseudomonas spp. growth.

High-Dose Neutron Detector Development Using 10B Coated Cells

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

Menlove, Howard Olsen; Henzlova, Daniela

2016-11-08

During FY16 the boron-lined parallel-plate technology was optimized to fully benefit from its fast timing characteristics in order to enhance its high count rate capability. To facilitate high count rate capability, a novel fast amplifier with timing and operating properties matched to the detector characteristics was developed and implemented in the 8” boron plate detector that was purchased from PDT. Each of the 6 sealed-cells was connected to a fast amplifier with corresponding List mode readout from each amplifier. The FY16 work focused on improvements in the boron-10 coating materials and procedures at PDT to significantly improve the neutron detectionmore » efficiency. An improvement in the efficiency of a factor of 1.5 was achieved without increasing the metal backing area for the boron coating. This improvement has allowed us to operate the detector in gamma-ray backgrounds that are four orders of magnitude higher than was previously possible while maintaining a relatively high counting efficiency for neutrons. This improvement in the gamma-ray rejection is a key factor in the development of the high dose neutron detector.« less

Microbiological quality and potential public health risks of export meat from springbok (Antidorcas marsupialis) in Namibia.

PubMed

Magwedere, K; Shilangale, R; Mbulu, R S; Hemberger, Y; Hoffman, L C; Dziva, F

2013-01-01

To assess the microbiological quality and safety of export game meat; i) a total of 80 pooled meat samples for aerobic plate count (APC) and Enterobacteriaceae ii) water used in harvesting and processing for microbiological quality and iii) meat and rectal contents for Salmonella spp. and Shiga toxin Escherichia coli (STEC) were evaluated in 2009 and 2010. No differences (p>0.05) in the APCs were observed between the years, but the mean Enterobacteriaceae count for 2009 was 1.33 ± 0.69 log(10)cfu/cm(2) compared to 2.93 ± 1.50 log(10)cfu/cm(2) for 2010. Insignificant Heterotrophic Plate Count (HPC) levels were detected in 9/23 field water samples, while fecal bacterial (coliforms, Clostridium perfringens and enterococci) were absent in all samples. No Salmonella spp. was isolated and all E. coli isolates from meat were negative for STEC virulence genes (stx1, stx2, eae and hlyA), suggesting a negligible role by springbok in the epidemiology of STEC and Salmonella. Copyright © 2012 Elsevier Ltd. All rights reserved.

High precision refractometry based on Fresnel diffraction from phase plates.

PubMed

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

2012-05-01

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

Evaluation of variability and quality control procedures for a receptor-binding assay for paralytic shellfish poisoning toxins.

PubMed

Ruberu, S R; Langlois, G W; Masuda, M; Perera, S Kusum

2012-01-01

The receptor-binding assay (RBA) method for determining saxatoxin (STX) and its numerous analogues, which cause paralytic shellfish poisoning (PSP) in humans, was evaluated in a single laboratory study. Each step of the assay preparation procedure including the performance of the multi-detector TopCount® instrument was evaluated for its contribution to method variability. The overall inherent RBA variability was determined to be 17%. Variability within the 12 detectors was observed; however, there was no reproducible pattern in detector performance. This observed variability among detectors could be attributed to other factors, such as pipetting errors. In an attempt to reduce the number of plates rejected due to excessive variability in the method's quality control parameters, a statistical approach was evaluated using either Grubbs' test or the Student's t-test for rejecting outliers in the measurement of triplicate wells. This approach improved the ratio of accepted versus rejected plates, saving cost and time for rerunning the assay. However, the potential reduction in accuracy and the lack of improvement in precision suggests caution when using this approach. The current study has recommended an alternate quality control procedure for accepting or rejecting plates in place of the criteria currently used in the published assay, or the alternative of outlier testing. The recommended procedure involves the development of control charts to monitor the critical parameters identified in the published method (QC sample, EC₅₀, slope of calibration curve), with the addition of a fourth critical parameter which is the top value (100% binding) of the calibration curve.

New procedure to reduce the time and cost of broncho-pulmonary specimen management using the Previ Isola® automated inoculation system.

PubMed

Nebbad-Lechani, Biba; Emirian, Aurélie; Maillebuau, Fabienne; Mahjoub, Nadia; Fihman, Vincent; Legrand, Patrick; Decousser, Jean-Winoc

2013-12-01

The microbiological diagnosis of respiratory tract infections requires serial manual dilutions of the clinical specimen before agar plate inoculation, disrupting the workflow in bacteriology clinical laboratories. Automated plating instrument systems have been designed to increase the speed, reproducibility and safety of this inoculating step; nevertheless, data concerning respiratory specimens are lacking. We tested a specific procedure that uses the Previ Isola® (bioMérieux, Craponne, France) to inoculate with broncho-pulmonary specimens (BPS). A total of 350 BPS from a university-affiliated hospital were managed in parallel using the manual reference and the automated methods (expectoration: 75; broncho-alveolar lavage: 68; tracheal aspiration: 17; protected distal sample: 190). A specific enumeration reading grid, a pre-liquefaction step and a fluidity test, performed before the inoculation, were designed for the automated method. The qualitative (i.e., the number of specimens yielding a bacterial count greater than the clinical threshold) and quantitative (i.e., the discrepancy within a 0.5 log value) concordances were 100% and 98.2%, respectively. The slimmest subgroup of expectorations could not be managed by the automated method (8%, 6/75). The technical time and cost savings (i.e., number of consumed plates) reached 50%. Additional studies are required for specific populations, such as cystic fibrosis specimens and associated bacterial variants. An automated decapper should be implemented to increase the biosafety of the process. The PREVI Isola® adapted procedure is a time- and cost-saving method for broncho-pulmonary specimen processing. © 2013.

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

NASA Astrophysics Data System (ADS)

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.

UV inactivation of pathogenic and indicator microorganisms

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

Chang, J.C.; Ossoff, S.F.; Lobe, D.C.

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 4more » 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.« less

Assessment of the bacteriological activity associated with granular activated carbon treatment of drinking water.

PubMed Central

Stewart, M H; Wolfe, R L; Means, E G

1990-01-01

Bacteriological analyses were performed on the effluent from a conventional water treatment pilot plant in which granular activated carbon (GAC) had been used as the final process to assess the impact of GAC on the microbial quality of the water produced. Samples were collected twice weekly for 160 days from the effluents of six GAC columns, each of which used one of four different empty-bed contact times (7.5, 15, 30, and 60 min). The samples were analyzed for heterotrophic plate counts and total coliforms. Effluent samples were also exposed to chloramines and free chlorine for 60 min (pH 8.2, 23 degrees C). Bacterial identifications were performed on the disinfected and nondisinfected effluents. Additional studies were conducted to assess the bacteriological activity associated with released GAC particles. The results indicated that heterotrophic plate counts in the effluents from all columns increased to 10(5) CFU/ml within 5 days and subsequently stabilized at 10(4) CFU/ml. The heterotrophic plate counts did not differ at different empty-bed contact times. Coliforms (identified as Enterobacter spp.) were recovered from the nondisinfected effluent on only two occasions. The disinfection results indicated that 1.5 mg of chloramines per liter inactivated approximately 50% more bacteria than did 1.0 mg of free chlorine per liter after 1 h of contact time. Chloramines and chlorine selected for the development of different bacterial species--Pseudomonas spp. and Flavobacterium spp., respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2082828

Microorganisms associated with production lots of the nucleopolyhedrosis virus of the gypsy moth Lymantria dispar (Lep.: Lymantriidae)

Treesearch

J.D. Podgwaite; R.B. Bruen; M. Shapiro

1983-01-01

Samples of a gypsy moth nucleopolyhedrosis virus product, Gypchek®, were taken each day during a 100-day production run and monitored for the presence of pathogenic bacteria and fungi. The standard plate count/g of product was 5.97 ± 1.51 x 108 over the 100-day period, while the sporulating bacteria count was 3.81 ± 1.21 x 106...

Transforming GSC-II Magnitudes into JWST/FGS Count Rates

NASA Astrophysics Data System (ADS)

Holfeltz, Sherie T.; Chayer, P.; Nelan, E. P.

2010-01-01

The JWST Fine Guidance Sensor (FGS) will provide the positions of guide stars to the spacecraft attitude control system to facilitate the fine pointing of the Observatory. The FGS is an infrared camera operating in an unfiltered passband from 0.6 to 5.3 microns. The ground system will select guide stars from the Guide Star Catalog II (GSC-II), which is an all-sky catalog with three optical passbands (BJ, RF, IN) derived from photographic plates, and from 2MASS. We present a method for predicting a guide star's FGS photon count rate, which is needed to operate the FGS. The method consists of first deriving equations for transforming the GSC-II optical passbands into J, H, and K for stars that are below the 2MASS faint limiting magnitude, based upon fitting the distribution of brighter stars in color-color diagrams using GSC-II and 2MASS photometry. Next, we convolve the BJ, RF, IN and predicted J, H, and K magnitudes (or 2MASS magnitudes if available) for a given star with the wavelength dependent throughput and sensitivity of the telescope and FGS. To estimate the accuracy of this method for stars that are too faint for 2MASS, we compare the predicted J, H, and K magnitudes for a large sample of stars to data from the United Kingdom Infrared Telescope (UKIRT) Deep Sky Survey (UKIDSS) Large Area Survey (LAS). Using synthetic magnitudes computed from Kurucz models for stars of different spectral types, we show that the method should provide reliable FGS count rates.

GreenLight Model 960.

PubMed

Fernandes, Richard; Carey, Conn; Hynes, James; Papkovsky, Dmitri

2013-01-01

The importance of food safety has resulted in a demand for a more rapid, high-throughput method for total viable count (TVC). The industry standard for TVC determination (ISO 4833:2003) is widely used but presents users with some drawbacks. The method is materials- and labor-intensive, requiring multiple agar plates per sample. More importantly, the method is slow, with 72 h typically required for a definitive result. Luxcel Biosciences has developed the GreenLight Model 960, a microtiter plate-based assay providing a rapid high-throughput method of aerobic bacterial load assessment through analysis of microbial oxygen consumption. Results are generated in 1-12 h, depending on microbial load. The mix and measure procedure allows rapid detection of microbial oxygen consumption and equates oxygen consumption to microbial load (CFU/g), providing a simple, sensitive means of assessing the microbial contamination levels in foods (1). As bacteria in the test sample grow and respire, they deplete O2, which is detected as an increase in the GreenLight probe signal above the baseline level (2). The time required to reach this increase in signal can be used to calculate the CFU/g of the original sample, based on a predetermined calibration. The higher the initial microbial load, the earlier this threshold is reached (1).

Efficacy of hypobromous acid as a hide-on carcass antimicrobial intervention.

PubMed

Schmidt, John W; Wang, Rong; Kalchayanand, Norasak; Wheeler, Tommy L; Koohmaraie, Mohammad

2012-05-01

Escherichia coli O157:H7 and Salmonella on cattle hides at slaughter are the main source of beef carcass contamination by these foodborne pathogens during processing. Hypobromous acid (HOBr) has been approved for various applications in meat processing, but the efficacy of HOBr as a hide antimicrobial has not been determined. In this study, the antimicrobial properties of HOBr were determined by spraying cattle hides at either of two concentrations, 220 or 500 ppm. Treatment of hides with 220 ppm of HOBr reduced the prevalence of E. coli O157:H7 on hides from 25.3 to 10.1% (P < 0.05) and reduced the prevalence of Salmonella from 28.3 to 7.1% (P < 0.05). Treatment of hides with 500 ppm of HOBr reduced (P < 0.05) the prevalence of E. coli O157:H7 on hides from 21.2 to 10.1% and the prevalence of Salmonella from 33.3 to 8.1%. The application of 220 ppm of HOBr reduced (P < 0.05) aerobic plate counts, total coliform counts, and E. coli counts on hides by 2.2 log CFU/ 100 cm(2). The use of 500 ppm of HOBr resulted in reductions (P < 0.05) of aerobic plate counts, total coliform counts, and E. coli counts by 3.3, 3.7, and 3.8 log CFU/100 cm(2), respectively, demonstrating that the use of higher concentrations of HOBr on hides resulted in additional antimicrobial activity. These results indicate that the adoption of a HOBr hide wash will reduce hide concentrations of spoilage bacteria and pathogen prevalence, resulting in a lower risk of carcass contamination.

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

PubMed

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

2012-03-01

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

Bedload transport rates in a gravel bedded-river derived from high-resolution monitoring using seismic impact plates

NASA Astrophysics Data System (ADS)

Downs, Peter; Soar, Philip

2015-04-01

Accurate characterisation of bedload transport rates is critical for a better understanding of geomorphological process dynamics, aquatic habitats, sediment budgets and strategies for catchment-scale initiatives in sediment management under conditions of climate change. However, rate estimation is challenging in practice: direct measurements are costly and logistically difficult to achieve with acceptable accuracy over geomorphologically-relevant time periods, and the uncertainty in transport rates predicted from empirical formulae and numerical simulation is rarely below 50 per cent. Partly reflecting these issues, passive technologies for continuous bedload monitoring are becoming increasingly popular. Sensors such as seismic impact plates offer the opportunity to characterise bedload activity at exceptionally high resolution - monitoring from the River Avon, (Devon, UK) indicated that despite significant intra-event and between-plate differences in apparent bedload transport aggregated over 5-minute periods, the magnitude-frequency product of discharge and impact frequency result in a highly plausible effective discharge, supporting the potential value of impact plates as indicators of relative sediment transport loads over annual timescales. Whereas the focus in bedload rate estimation to date has been on developing satisfactory sediment rating curves from detection signals, we instead develop a method for directly estimating bedload transport rates from impact plate data as a function of intensity of transport (count, n, per second), bed material mass (kg) and cross-stream transport variability. Bulk sediment samples are converted to a mass in transit for each instantaneous discharge according to the intensity of transport and a Monte Carlo simulation of the load in transit determined at random from the bed material particle size distribution. The lower detection threshold is determined using experimental calibration and the upper size limit is determined from incipient motion estimates thereby establishing the fraction of transported material sensed by the plates. The lateral variability in transport rates across the cross-section is estimated empirically using multiple plates or by interpolation. This procedure provides a potentially affordable and robust method of achieving uncertainty-bound indicative measures of bedload transport with the potential for wide-ranging practical applications.

Calorie count - sodas and energy drinks

MedlinePlus

... Accessed May 11, 2016. United States Department of Agriculture website. ChooseMyPlate.gov. Make better beverage choices. www. ... Accessed May 11, 2016. United States Department of Agriculture. National nutrient database for standard reference. Release 28. ...

7 CFR 58.653 - Microbiological requirements for sherbet.

Code of Federal Regulations, 2011 CFR

2011-01-01

... requirements for sherbet. The finished product shall contain not more than 50,000 bacteria per gram as determined by the standard plate count and shall contain not more than 10 coliform organisms per gram in...

7 CFR 58.653 - Microbiological requirements for sherbet.

Code of Federal Regulations, 2010 CFR

2010-01-01

... requirements for sherbet. The finished product shall contain not more than 50,000 bacteria per gram as determined by the standard plate count and shall contain not more than 10 coliform organisms per gram in...

7 CFR 58.653 - Microbiological requirements for sherbet.

Code of Federal Regulations, 2013 CFR

2013-01-01

... requirements for sherbet. The finished product shall contain not more than 50,000 bacteria per gram as determined by the standard plate count and shall contain not more than 10 coliform organisms per gram in...

Comprehensive System-Based Architecture for an Integrated High Energy Laser Test Bed

DTIC Science & Technology

2015-03-01

76 4. Comparison of Sensors ................................................................76 B. TRANSMISSION...81 b. Photometers .......................................................................84 4. Comparison of Sensors ...88 a. Flat Plate Target Boards, Ablatives, and Acrylite ...........88 b. Photon-Counting Sensors

Rapid Quantitative Detection of Lactobacillus sakei in Meat and Fermented Sausages by Real-Time PCR

PubMed Central

Martín, Belén; Jofré, Anna; Garriga, Margarita; Pla, Maria; Aymerich, Teresa

2006-01-01

A quick and simple method for quantitative detection of Lactobacillus sakei in fermented sausages was successfully developed. It is based on Chelex-100-based DNA purification and real-time PCR enumeration using a TaqMan fluorescence probe. Primers and probes were designed in the L. sakei 16S-23S rRNA intergenic transcribed spacer region, and the assay was evaluated using L. sakei genomic DNA and an artificially inoculated sausage model. The detection limit of this technique was approximately 3 cells per reaction mixture using both purified DNA and the inoculated sausage model. The quantification limit was established at 30 cells per reaction mixture in both models. The assay was then applied to enumerate L. sakei in real samples, and the results were compared to the MRS agar count method followed by confirmation of the percentage of L. sakei colonies. The results obtained by real-time PCR were not statistically significantly different than those obtained by plate count on MRS agar (P > 0.05), showing a satisfactory agreement between both methods. Therefore, the real-time PCR assay developed can be considered a promising rapid alternative method for the quantification of L. sakei and evaluation of the implantation of starter strains of L. sakei in fermented sausages. PMID:16957227

The Rebirth of Culture in Microbiology through the Example of Culturomics To Study Human Gut Microbiota

PubMed Central

Lagier, Jean-Christophe; Hugon, Perrine; Khelaifia, Saber; Fournier, Pierre-Edouard; La Scola, Bernard

2015-01-01

SUMMARY Bacterial culture was the first method used to describe the human microbiota, but this method is considered outdated by many researchers. Metagenomics studies have since been applied to clinical microbiology; however, a “dark matter” of prokaryotes, which corresponds to a hole in our knowledge and includes minority bacterial populations, is not elucidated by these studies. By replicating the natural environment, environmental microbiologists were the first to reduce the “great plate count anomaly,” which corresponds to the difference between microscopic and culture counts. The revolution in bacterial identification also allowed rapid progress. 16S rRNA bacterial identification allowed the accurate identification of new species. Mass spectrometry allowed the high-throughput identification of rare species and the detection of new species. By using these methods and by increasing the number of culture conditions, culturomics allowed the extension of the known human gut repertoire to levels equivalent to those of pyrosequencing. Finally, taxonogenomics strategies became an emerging method for describing new species, associating the genome sequence of the bacteria systematically. We provide a comprehensive review on these topics, demonstrating that both empirical and hypothesis-driven approaches will enable a rapid increase in the identification of the human prokaryote repertoire. PMID:25567229

Rapid quantitative detection of Lactobacillus sakei in meat and fermented sausages by real-time PCR.

PubMed

Martín, Belén; Jofré, Anna; Garriga, Margarita; Pla, Maria; Aymerich, Teresa

2006-09-01

A quick and simple method for quantitative detection of Lactobacillus sakei in fermented sausages was successfully developed. It is based on Chelex-100-based DNA purification and real-time PCR enumeration using a TaqMan fluorescence probe. Primers and probes were designed in the L. sakei 16S-23S rRNA intergenic transcribed spacer region, and the assay was evaluated using L. sakei genomic DNA and an artificially inoculated sausage model. The detection limit of this technique was approximately 3 cells per reaction mixture using both purified DNA and the inoculated sausage model. The quantification limit was established at 30 cells per reaction mixture in both models. The assay was then applied to enumerate L. sakei in real samples, and the results were compared to the MRS agar count method followed by confirmation of the percentage of L. sakei colonies. The results obtained by real-time PCR were not statistically significantly different than those obtained by plate count on MRS agar (P > 0.05), showing a satisfactory agreement between both methods. Therefore, the real-time PCR assay developed can be considered a promising rapid alternative method for the quantification of L. sakei and evaluation of the implantation of starter strains of L. sakei in fermented sausages.

Microchannel plate EUV detectors for the Extreme Ultraviolet Explorer

NASA Technical Reports Server (NTRS)

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

1984-01-01

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

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

NASA Astrophysics Data System (ADS)

Akindinov, A.; Dreyer, J.; Fan, X.; Kämpfer, B.; Kiselev, S.; Kotte, R.; Garcia, A. Laso; Malkevich, D.; Naumann, L.; Nedosekin, A.; Plotnikov, V.; Stach, D.; Sultanov, R.; Voloshin, K.

2017-02-01

Resistive Plate Chambers with ceramic electrodes are the main candidates for a use in precise multi-channel timing systems operating in high-radiation conditions. We report the latest R&D results on these detectors aimed to meet the requirements of the forward T0 counter at the CBM experiment. RPC design, gas mixture, limits on the bulk resistivity of ceramic electrodes, efficiency, time resolution, counting rate capabilities and ageing test results are presented.

A homogeneous method to measure aminoacyl-tRNA synthetase aminoacylation activity using scintillation proximity assay technology.

PubMed

Macarrón, R; Mensah, L; Cid, C; Carranza, C; Benson, N; Pope, A J; Díez, E

2000-09-10

A new method to measure the aminoacylation of tRNA based upon the use of the scintillation proximity assay (SPA) technology has been developed. The assay detects incorporation of radiolabeled amino acids into cognate tRNA, catalyzed by a specific aminoacyl-tRNA synthetase (aaRS). Under acidic conditions, uncoated yttrium silicate SPA beads were found to bind tRNA aggregates, while the radiolabeled amino acid substrate remains in solution, resulting in good signal discrimination of these two species in the absence of any separation steps. The usefulness of this approach was demonstrated by measurement of steady-state kinetic constants and inhibitor binding constants for a range of aaRS enzymes in comparison with data from standard, trichloroacetic acid-precipitation-based assays. In all cases, the data were quantitatively comparable. Although the radioisotopic counting efficiency of the SPA method was less than that of standard liquid scintillation counting, the statistical performance (i.e., signal to background, variability, stability) of the SPA assays was at least equivalent to the separation-based methods. The assay was also shown to work well in miniaturized 384-well microtiter plate formats, resulting in considerable reagent savings. In summary, a new method to characterize aaRS activity is described that is faster and more amenable to high-throughput screening than traditional methods. Copyright 2000 Academic Press.

Protective isolation in single-bed rooms: studies in a modified hospital ward

PubMed Central

Ayliffe, G. A. J.; Collins, B. J.; Lowbury, E. J. L.; Wall, Mary

1971-01-01

Studies were made in a modified hospital ward containing 19 beds, 14 of them in the open ward, one in a window-ventilated side-room, two in rooms with partial-recirculation ventilators giving 7-10 air changes per hour, and two in self-contained isolation suites with plenum ventilation (20 air changes per hour), ultra-violet (UV) barriers at doorways and airlocks. Preliminary tests with aerosols of tracer bacteria showed that few bacteria entered the plenum or recirculation-ventilated rooms. Bacteria released inside mechanically ventilated cubicles escaped into the corridor, but this transfer was reduced by the presence of an airlock. UV barriers at the entrance to the airlock and the cubicle reduced the transfer of bacteria from cubicle to corridor. During a period of 4 years while the ward was in use for surgical and gynaecological patients, the incidence of post-operative sepsis and colonization of wounds by multiple-resistant Staphylococcus aureus was lower (though not significantly lower) in the plenum-ventilated rooms than in the open ward, the recirculator-ventilated cubicles and the window-ventilated cubicles. Nasal acquisition of multiple-resistant Staph. aureus was significantly less common in the plenum-ventilated than in the recirculator-ventilated cubicles and in the other areas. Mean counts of bacteria on settle-plates were significantly lower in the plenum-ventilated cubicles than in the other areas; mean settle-plate counts in the recirculator-ventilated cubicles were significantly lower than in the open ward and in the window-ventilated side-room; similar results were shown by slit-sampling of air. Mean settle-plate counts were significantly lower in all areas when the ward was occupied by female patients. Staph. aureus was rarely carried by air from plenum-ventilated or other cubicles to the open ward, or from the open ward to the cubicles; though staphylococci were transferred from one floor area to another, they did not appear to be redispersed into the air in sufficient numbers to infect the patients. Ultra-violet irradiation caused a significant reduction in the total and staphylococcal counts from the floors of airlocks, and a significant reduction of total counts in the air. PMID:5289715

Noise-free accurate count of microbial colonies by time-lapse shadow image analysis.

PubMed

Ogawa, Hiroyuki; Nasu, Senshi; Takeshige, Motomu; Funabashi, Hisakage; Saito, Mikako; Matsuoka, Hideaki

2012-12-01

Microbial colonies in food matrices could be counted accurately by a novel noise-free method based on time-lapse shadow image analysis. An agar plate containing many clusters of microbial colonies and/or meat fragments was trans-illuminated to project their 2-dimensional (2D) shadow images on a color CCD camera. The 2D shadow images of every cluster distributed within a 3-mm thick agar layer were captured in focus simultaneously by means of a multiple focusing system, and were then converted to 3-dimensional (3D) shadow images. By time-lapse analysis of the 3D shadow images, it was determined whether each cluster comprised single or multiple colonies or a meat fragment. The analytical precision was high enough to be able to distinguish a microbial colony from a meat fragment, to recognize an oval image as two colonies contacting each other, and to detect microbial colonies hidden under a food fragment. The detection of hidden colonies is its outstanding performance in comparison with other systems. The present system attained accuracy for counting fewer than 5 colonies and is therefore of practical importance. Copyright © 2012 Elsevier B.V. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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

2014-07-01

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

Improved soft-agar colony assay in a fluid processing apparatus.

PubMed

Forsman, A D; Herpich, A R; Chapes, S K

1999-01-01

The standard method for quantitating bone marrow precursor cells has been to count the number of colony-forming units that form in semisolid (0.3%) agar. Recently we adapted this assay for use in hardware, the Fluid Processing Apparatus, that is flown in standard payload lockers of the space shuttle. When mouse or rat macrophage colony-forming units were measured with this hardware in ground-based assays, we found significantly more colony growth than that seen in standard plate assays. The improved growth correlates with increased agar thickness but also appears to be due to properties inherent to the Fluid Processing Apparatus. This paper describes an improved method for determining bone marrow macrophage precursor numbers in semisolid agar.

Effect of extrusion processing on the microstructure, mechanical properties, biocorrosion properties and antibacterial properties of Ti-Cu sintered alloys.

PubMed

Zhang, Erlin; Li, Shengyi; Ren, Jing; Zhang, Lan; Han, Yong

2016-12-01

Ti-Cu sintered alloys, Ti-Cu(S) alloy, have exhibited good anticorrosion resistance and strong antibacterial properties, but low ductility in previous study. In this paper, Ti-Cu(S) alloys were subjected to extrusion processing in order to improve the comprehensive property. The phase constitute, microstructure, mechanical property, biocorrosion property and antibacterial activity of the extruded alloys, Ti-Cu(E), were investigated in comparison with Ti-Cu(S) by X-ray diffraction (XRD), optical microscopy (OM), scanning electronic microscopy (SEM) with energy disperse spectroscopy (EDS), mechanical testing, electrochemical testing and plate-count method in order to reveal the effect of the extrusion process. XRD, OM and SEM results showed that the extrusion process did not change the phase constitute but refined the grain size and Ti2Cu particle significantly. Ti-Cu(E) alloys exhibited higher hardness and compressive yield strength than Ti-Cu(S) alloys due to the fine grain and Ti2Cu particles. With the consideration of the total compressive strain, it was suggested that the extrusion process could improve the ductility of Ti-Cu alloy(S) alloys. Electrochemical results have indicated that the extrusion process improved the corrosion resistance of Ti-Cu(S) alloys. Plate-count method displayed that both Ti-Cu(S) and Ti-Cu(E) exhibited strong antibacterial activity (>99%) against S. aureus. All these results demonstrated that hot forming processing, such as the extrusion in this study, refined the microstructure and densified the alloy, in turn improved the ductility and strength as well as anticorrosion properties without reduction in antibacterial properties. Copyright © 2016 Elsevier B.V. All rights reserved.

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

PubMed

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

2005-10-01

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

Lung counting: comparison of detector performance with a four detector array that has either metal or carbon fibre end caps, and the effect on mda calculation.

PubMed

Ahmed, Asm Sabbir; Hauck, Barry; Kramer, Gary H

2012-08-01

This study described the performance of an array of high-purity Germanium detectors, designed with two different end cap materials-steel and carbon fibre. The advantages and disadvantages of using this detector type in the estimation of the minimum detectable activity (MDA) for different energy peaks of isotope (152)Eu were illustrated. A Monte Carlo model was developed to study the detection efficiency for the detector array. A voxelised Lawrence Livermore torso phantom, equipped with lung, chest plates and overlay plates, was used to mimic a typical lung counting protocol with the array of detectors. The lung of the phantom simulated the volumetric source organ. A significantly low MDA was estimated for energy peaks at 40 keV and at a chest wall thickness of 6.64 cm.

In vivo evaluation of the effect of essential oil-containing oral strips on salivary bacteria using the checkerboard method.

PubMed

da Silva, Carina Maciel; Colombo, Andréa Vieira; do Souto, Renata Martins; Colombo, Ana Paula

2005-01-01

The aim of this study was to evaluate the antimicrobial effect of essential oil-containing oral strips on different species of the oral microbiota. Saliva samples were collected from 20 subjects with good oral health, diluted and plated onto blood agar medium. The subjects were asked to place the strip (Listerine PocketPaks) on the tongue allowing it to dissolve. After 30 minutes, new saliva samples were collected again and the plates with the samples were incubated under anaerobic conditions at 37 degrees C for seven days. Colony counts (CFU/mL) were determined for each sample. The colonies on the plates were washed with 1 mL of TE buffer, and the bacterial suspensions were processed for the identification of 24 species by DNA probes and the Checkerboard DNA-DNA hybridization method. Differences in total counts, prevalence, and levels of the species evaluated before and after placement of the strips were determined by Wilcoxon sign rank and Chi-square tests. A modest increase in the total bacterial number in saliva from 1.4 x 10(8) to 1.7 x 10(8) bacterial cells was observed 30 minutes after the strip placement, although this change was not significant (p = 0.632). Most of the species reduced in frequency and/or levels, including the pathogens A. actinomycetemcomitans, C. rectus, E. corrodens, Fusobacterium spp., P. intermedia, and S. noxia, as well as the beneficial species A. meyeri, A. georgia, A. gerencseriae, A. odontolyticus, and P. acnes after strip placement. In contrast, A. viscosus, P. melaninogenica, P. gingivalis, P. micros, Streptococcus spp., T. forsythensis, and V. parvula presented an increase in prevalence and/or levels. These changes were not statistically significant after adjusting for multiple comparisons (p > 0.0022). The use of the essential oil-containing oral strips resulted in a short-term small increase in the total number of salivary microorganisms. In addition, a not significant decrease of certain periodontopathogens, and an increase in species compatible with oral health were observed.

Spatial distribution of Legionella pneumophila MLVA-genotypes in a drinking water system.

PubMed

Rodríguez-Martínez, Sarah; Sharaby, Yehonatan; Pecellín, Marina; Brettar, Ingrid; Höfle, Manfred; Halpern, Malka

2015-06-15

Bacteria of the genus Legionella cause water-based infections, resulting in severe pneumonia. To improve our knowledge about Legionella spp. ecology, its prevalence and its relationships with environmental factors were studied. Seasonal samples were taken from both water and biofilm at seven sampling points of a small drinking water distribution system in Israel. Representative isolates were obtained from each sample and identified to the species level. Legionella pneumophila was further determined to the serotype and genotype level. High resolution genotyping of L. pneumophila isolates was achieved by Multiple-Locus Variable number of tandem repeat Analysis (MLVA). Within the studied water system, Legionella plate counts were higher in summer and highly variable even between adjacent sampling points. Legionella was present in six out of the seven selected sampling points, with counts ranging from 1.0 × 10(1) to 5.8 × 10(3) cfu/l. Water counts were significantly higher in points where Legionella was present in biofilms. The main fraction of the isolated Legionella was L. pneumophila serogroup 1. Serogroup 3 and Legionella sainthelensis were also isolated. Legionella counts were positively correlated with heterotrophic plate counts at 37 °C and negatively correlated with chlorine. Five MLVA-genotypes of L. pneumophila were identified at different buildings of the sampled area. The presence of a specific genotype, "MLVA-genotype 4", consistently co-occurred with high Legionella counts and seemed to "trigger" high Legionella counts in cold water. Our hypothesis is that both the presence of L. pneumophila in biofilm and the presence of specific genotypes, may indicate and/or even lead to high Legionella concentration in water. This observation deserves further studies in a broad range of drinking water systems to assess its potential for general use in drinking water monitoring and management. Copyright © 2015 Elsevier Ltd. All rights reserved.

A Novel 96well-formatted Micro-gap Plate Enabling Drug Response Profiling on Primary Tumour Samples

NASA Astrophysics Data System (ADS)

Ma, Wei-Yuan; Hsiung, Lo-Chang; Wang, Chen-Ho; Chiang, Chi-Ling; Lin, Ching-Hung; Huang, Chiun-Sheng; Wo, Andrew M.

2015-04-01

Drug-based treatments are the most widely used interventions for cancer management. Personalized drug response profiling remains inherently challenging with low cell count harvested from tumour sample. We present a 96well-formatted microfluidic plate with built-in micro-gap that preserves up to 99.2% of cells during multiple assay/wash operation and only 9,000 cells needed for a single reagent test (i.e. 1,000 cells per test spot x 3 selected concentration x triplication), enabling drug screening and compatibility with conventional automated workstations. Results with MCF7 and MDA-MB-231 cell lines showed that no statistical significance was found in dose-response between the device and conventional 96-well plate control. Primary tumour samples from breast cancer patients tested in the device also showed good IC50 prediction. With drug screening of primary cancer cells must consider a wide range of scenarios, e.g. suspended/attached cell types and rare/abundant cell availability, the device enables high throughput screening even for suspended cells with low cell count since the signature microfluidic cell-trapping feature ensures cell preservation in a multiple solution exchange protocol.

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

PubMed

Holah, J T; Hall, K E

2006-05-01

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

Microbial contamination of mobile phones in a health care setting in Alexandria, Egypt

PubMed Central

Selim, Heba Sayed; Abaza, Amani Farouk

2015-01-01

Aim: This study aimed at investigating the microbial contamination of mobile phones in a hospital setting. Methods: Swab samples were collected from 40 mobile phones of patients and health care workers at the Alexandria University Students’ Hospital. They were tested for their bacterial contamination at the microbiology laboratory of the High Institute of Public Health. Quantification of bacteria was performed using both surface spread and pour plate methods. Isolated bacterial agents were identified using standard microbiological methods. Methicillin-resistant Staphylococcus aureus was identified by disk diffusion method described by Bauer and Kirby. Isolated Gram-negative bacilli were tested for being extended spectrum beta lactamase producers using the double disk diffusion method according to the Clinical and Laboratory Standards Institute recommendations. Results: All of the tested mobile phones (100%) were contaminated with either single or mixed bacterial agents. The most prevalent bacterial contaminants were methicillin-resistant S. aureus and coagulase-negative staphylococci representing 53% and 50%, respectively. The mean bacterial count was 357 CFU/ml, while the median was 13 CFU/ml using the pour plate method. The corresponding figures were 2,192 and 1,720 organisms/phone using the surface spread method. Conclusions: Mobile phones usage in hospital settings poses a risk of transmission of a variety of bacterial agents including multidrug-resistant pathogens as methicillin-resistant S. aureus. The surface spread method is an easy and useful tool for detection and estimation of bacterial contamination of mobile phones. PMID:25699226

The Sydney University PAPA camera

NASA Astrophysics Data System (ADS)

Lawson, Peter R.

1994-04-01

The Precision Analog Photon Address (PAPA) camera is a photon-counting array detector that uses optical encoding to locate photon events on the output of a microchannel plate image intensifier. The Sydney University camera is a 256x256 pixel detector which can operate at speeds greater than 1 million photons per second and produce individual photon coordinates with a deadtime of only 300 ns. It uses a new Gray coded mask-plate which permits a simplified optical alignment and successfully guards against vignetting artifacts.

Comparison of Standard Culture-Based Method to Culture-Independent Method for Evaluation of Hygiene Effects on the Hand Microbiome.

PubMed

Zapka, C; Leff, J; Henley, J; Tittl, J; De Nardo, E; Butler, M; Griggs, R; Fierer, N; Edmonds-Wilson, S

2017-03-28

Hands play a critical role in the transmission of microbiota on one's own body, between individuals, and on environmental surfaces. Effectively measuring the composition of the hand microbiome is important to hand hygiene science, which has implications for human health. Hand hygiene products are evaluated using standard culture-based methods, but standard test methods for culture-independent microbiome characterization are lacking. We sampled the hands of 50 participants using swab-based and glove-based methods prior to and following four hand hygiene treatments (using a nonantimicrobial hand wash, alcohol-based hand sanitizer [ABHS], a 70% ethanol solution, or tap water). We compared results among culture plate counts, 16S rRNA gene sequencing of DNA extracted directly from hands, and sequencing of DNA extracted from culture plates. Glove-based sampling yielded higher numbers of unique operational taxonomic units (OTUs) but had less diversity in bacterial community composition than swab-based sampling. We detected treatment-induced changes in diversity only by using swab-based samples ( P < 0.001); we were unable to detect changes with glove-based samples. Bacterial cell counts significantly decreased with use of the ABHS ( P < 0.05) and ethanol control ( P < 0.05). Skin hydration at baseline correlated with bacterial abundances, bacterial community composition, pH, and redness across subjects. The importance of the method choice was substantial. These findings are important to ensure improvement of hand hygiene industry methods and for future hand microbiome studies. On the basis of our results and previously published studies, we propose recommendations for best practices in hand microbiome research. IMPORTANCE The hand microbiome is a critical area of research for diverse fields, such as public health and forensics. The suitability of culture-independent methods for assessing effects of hygiene products on microbiota has not been demonstrated. This is the first controlled laboratory clinical hand study to have compared traditional hand hygiene test methods with newer culture-independent characterization methods typically used by skin microbiologists. This study resulted in recommendations for hand hygiene product testing, development of methods, and future hand skin microbiome research. It also demonstrated the importance of inclusion of skin physiological metadata in skin microbiome research, which is atypical for skin microbiome studies. Copyright © 2017 Zapka et al.

A Microbial Community in Sediments Beneath the Western Antarctic Ice Sheet, Ice Stream C (Kamb)

NASA Astrophysics Data System (ADS)

Skidmore, M.; Han, S.; Foo, W.; Bui, D.; Lanoil, B.

2004-12-01

In 2000, an ice-drilling project focusing on the "sticky spot" of Ice Stream C recovered cores of sub-glacial sediments from beneath the Western Antarctic Ice Sheet. We have characterized several chemical and microbiological parameters of the sole intact sediment core. Pore waters extracted from these sediments were brackish and some were supersaturated with respect to calcite. Ion chromatography demonstrated the presence of several organic acids at low, but detectable, levels in the pore water. DAPI direct cell counts were approximately 107 cells g-1. Aerobic viable plate counts were much lower than direct cell counts; however, they were two orders of magnitude higher on plates incubated at low temperature (4 ° C; 3.63 x 105 CFU ml-1) than at higher temperatures (ca. 22° C; 1.5 x 103 CFU ml-1); no colonies were detected on plates incubated anaerobically at either temperature. 16S rDNA clone library analysis indicates extremely limited bacterial diversity in these samples: six phylogenetic clades were detected. The three dominant bacterial phylogenetic clades in the clone libraries (252 clones total) were most closely related to Thiobacillus thioparus (180 clones), Polaromonas vacuolata (34 clones), and Gallionella ferruginea (35 clones) and their relatives; one clone each represented the other three phylogenetic clades (most closely related to Ralstonia pickettii, Lysobacter antibioticus, and Xylella fastidiosa, respectively). These sequences match closely with sequences previously obtained from other subglacial environments in Alaska, Ellesmere Island, Canada and New Zealand. Implications of this microbial community to subglacial chemistry and microbial biogeography will be discussed.

Evaluation of the repeatability of a crude adult indirect Ostertagia ostertagi ELISA and methods of expressing test results.

PubMed

Sanchez, J; Dohoo, I R; Markham, F; Leslie, K; Conboy, G

2002-10-16

An indirect enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against Ostertagia ostertagi using a crude adult worm antigen was evaluated using serum and milk samples from adult cows, as well as from bulk tank milk. Within and between plate repeatabilities were determined. In addition, the effects of factors such as antigen batch, freezing, preserving of the samples and somatic cell counts (SCCs) of the samples were evaluated. Raw optical densities (ODs) and normalized values were compared using the concordance correlation coefficient (CCC), the coefficient of variation (CV), Bland-Altman plots (BA). Based on raw OD values, there was a high repeatability within a plate (CCC approximately 0.96 and CV<10%). Repeatability between plates was evaluated following normalization of OD values by four methods. Computing normalized values as (OD-Nt)/(Pst-Nt), gave the most repeatable results, with the CCC being approximately 0.95 and the CV approximately 11%. When the OD values were higher than 1.2 and 0.3 for the positive and the negative controls, respectively, none of the normalization methods evaluated provided highly repeatable results and it was necessary to repeat the test. Two batches of the crude antigen preparation were evaluated for repeatability, and no difference was found (CCC=0.96). The use of preservative (bronopol) did not affect test results, nor did freezing the samples for up to 8 months. A significant positive relationship between ELISA OD for milk samples and SCC score was found. Therefore, the use of composite milk samples, which have less variable SCC than samples taken from each quarter, would be more suitable when the udder health status is unknown. The analytical methods used to evaluate repeatability provided a practical way to select among normalization procedures.

Bacteriological quality of drinking water from dispensers (coolers) and possible control measures.

PubMed

Baumgartner, Andreas; Grand, Marius

2006-12-01

Three water dispensers (coolers) were bacteriologically monitored over a period of 3 months to evaluate their hygienic status. For this purpose, 174 samples of chilled and unchilled water were analyzed for levels of mesophilic aerobic bacteria and the presence of Escherichia coli and enterococci in 100-ml samples, and the presence of Pseudomonas aeruginosa in 10- and 100-ml samples. Additionally, 12 samples from 20-liter plastic bottles of spring water used to supply the coolers and 36 samples of 12 different brands of noncarbonated bottled mineral water were similarly analyzed. Water from the coolers yielded aerobic plate counts of 3 to 5 log CFU/ml with a geometric mean of 3.86 log CFU/ml, whereas water from the 20-liter bottles had a mean aerobic plate count of 3.3 log CFU/ml. Aerobic plate counts for noncarbonated mineral waters were generally lower (13 samples, < 10 CFU/ml; 6 samples, 10 to 10(2) CFU/ml; 13 samples, 10(2) to 10(3) CFU/ml; 3 samples, 10(3) to 10(4) CFU/ ml; 1 sample, 2 x 10(4) CFU/ml). Although occasional professional cleaning of the coolers did not affect the aerobic plate count, P. aeruginosa was successfully eliminated 2 weeks after cleaning, with only one cooler becoming recolonized. Neither E. coli nor enterococci was found in any of the water samples tested. However, P. aeruginosa was identified in three (25%) of twelve 100-ml samples from 20-liter bottles of spring water; a similar frequency of 24.1% was seen for water samples from coolers. Overall, 35 (21.6%) of 162 water samples (10 ml) from coolers also yielded P. aeruginosa, suggesting potential growth of P. aeruginosa in the dispensers. Pulsed-field gel electrophoresis typing and antibiotic susceptibility testing found 19 P. aeruginosa isolates from the coolers and bottles to be identical, indicating that a single strain originated from the bottled water rather than the surroundings of the coolers. Because P. aeruginosa can cause serious nosocomial infections, its spread should be strictly controlled in institutions caring for vulnerable people such as hospitals and nursing homes. Consequently, in keeping with legal requirement for bottled spring and mineral water in Switzerland, it is also advisable that P. aeruginosa be absent in 100-ml samples of cooler water.

The physical and microbiological quality of chicken meat in the different type of enterprise poultry slaughterhouse: a case study in Karanganyar District

NASA Astrophysics Data System (ADS)

Hertanto, B. S.; Nurmalasari, C. D. A.; Nuhriawangsa, A. M. P.; Cahyadi, M.; Kartikasari, L. R.

2018-01-01

The aim of this study was to determine the physical and microbiological quality of chicken meat produced by the different type of enterprise slaughterhouse in Karanganyar District. The number of 20 poultry slaughterhouses was determined by convenience sampling method. The samples of chicken meat were randomly collected from medium enterprise poultry slaughterhouses (n=12) and small enterprise poultry slaughterhouses (n=8). A survey was carried out among poultry slaughterhouses in Karanganyar District. All the samples were subjected to physical quality consisted of pH test, texture, and color, while microbiological quality consisted of total plate count, microbial detection of Escherichia coli and Salmonella. The data were analyzed using descriptive quantitative analysis. The study showed that chicken meat in 6 small enterprise slaughterhouses and 11 medium enterprise slaughterhouses had normal pH of 5.81 - 6.3. Color and texture of chicken meats had relatively normal in both small and medium enterprise slaughterhouses. The total plate count of chicken meat showed in both small and medium enterprise slaughterhouses was <1x106 CFU/gr. The test of bacterial contamination showed that 3 of small and medium enterprise slaughterhouses were positively contaminated by Escherichia coli of >1x101 CFU/gr, and Salmonella was detected in 1 medium enterprise slaughterhouse. The overall results of the study suggest that the potential risk of chicken meat contamination depends on the processing of chicken meat in poultry slaughterhouses.

Microbial contamination of mobile phones in a health care setting in Alexandria, Egypt.

PubMed

Selim, Heba Sayed; Abaza, Amani Farouk

2015-01-01

This study aimed at investigating the microbial contamination of mobile phones in a hospital setting. Swab samples were collected from 40 mobile phones of patients and health care workers at the Alexandria University Students' Hospital. They were tested for their bacterial contamination at the microbiology laboratory of the High Institute of Public Health. Quantification of bacteria was performed using both surface spread and pour plate methods. Isolated bacterial agents were identified using standard microbiological methods. Methicillin-resistant Staphylococcus aureus was identified by disk diffusion method described by Bauer and Kirby. Isolated Gram-negative bacilli were tested for being extended spectrum beta lactamase producers using the double disk diffusion method according to the Clinical and Laboratory Standards Institute recommendations. All of the tested mobile phones (100%) were contaminated with either single or mixed bacterial agents. The most prevalent bacterial contaminants were methicillin-resistant S. aureus and coagulase-negative staphylococci representing 53% and 50%, respectively. The mean bacterial count was 357 CFU/ml, while the median was 13 CFU/ml using the pour plate method. The corresponding figures were 2,192 and 1,720 organisms/phone using the surface spread method. Mobile phones usage in hospital settings poses a risk of transmission of a variety of bacterial agents including multidrug-resistant pathogens as methicillin-resistant S. aureus. The surface spread method is an easy and useful tool for detection and estimation of bacterial contamination of mobile phones.

Assessment of Plant-Probiotic Performance of Novel Endophytic Bacillus sp. in Talc-Based Formulation.

PubMed

Basheer, Jasim; Ravi, Aswani; Mathew, Jyothis; Krishnankutty, Radhakrishnan Edayileveettil

2018-01-25

Endophytic bacteria are considered to have a plethora of plant growth promoting and anti-phytopathogenic traits to live within the plants. Hence, they have immense promises for plant probiotic development. In the current study, plant probiotic endophytic Bacillus sp. CaB5 which has been previously isolated from Capsicum annuum was investigated for its performance in talc-based formulation. For this, CaB5 was made into formulation with sterile talc, calcium carbonate, and carboxymethyl cellulose. The viability analysis of the formulation by standard plate count and fluorescence methods has confirmed the stable microbial count up to 45 days. Plant probiotic performance of the prepared formulation was analyzed on cowpea (Vigna unguiculata) and lady's finger (Abelmoschus esculentus). The results showed the formulation treatment to have enhancement effect on seed germination as well as plant growth in both selected plants. The results highlight the potential of CaB5-based formulation for field application to enhance growth of economically important plants.

Applications of chemiluminescence to bacterial analysis

NASA Technical Reports Server (NTRS)

Searle, N. D.

1975-01-01

Luminol chemiluminescence method for detecting bacteria was based on microbial activation of the oxidation of the luminol monoanion by hydrogen peroxide. Elimination of the prior lysing step, previously used in the chemiluminescence technique, was shown to improve considerably the reproducibility and accuracy of the method in addition to simplifying it. An inexpensive, portable photomultiplier detector was used to measure the maximum light intensity produced when the sample is added to the reagent. Studies of cooling tower water show that the luminol chemiluminescence technique can be used to monitor changes in viable cell population both under normal conditions and during chlorine treatment. Good correlation between chemiluminescence and plate counts was also obtained in the analysis of process water used in paper mills. This method showed good potential for monitoring the viable bacteria populations in activated sludge used in waste treatment plants to digest organic matter.

Suppression of Listeria monocytogenes by the Native Micro-Flora in Teewurst Sausage.

PubMed

Austin-Watson, Clytrice; Grant, Ar'Quette; Brice, Michline

2013-10-21

Modern consumers are interested in the use of non-chemical methods to control pathogens when heat sterilization is not an option. Such is the case with teewurst sausage, a raw spreadable sausage and a popular German commodity. Although Listeria was not found in teewurst, the optimal microbial growing conditions of teewurst coupled with the ubiquity of L. monocytogenes in nature, makes the possibility of contamination of products very possible. This pilot study was conducted to examine teewurst's native micro-flora's ability to suppress the outgrowth of L. monocytogenes at 10 °C using standard plate counts and PCR-DGGE. Traditional plating methods showed L. monocytogenes growth significantly decreased when in competition with the teewurst's native micro-flora ( p < 0.05). The native micro-flora of the teewurst suppressed the overall growth of L. monocytogenes by an average of two logs, under these conditions. Denaturing Gradient Gel Electrophoresis (DGGE) amplicons with unique banding patterns were extracted from DGGE gel for identification. Brochothrix thermosphacta and Lactobacillus curvatus were identified as a part of the teewurst's native micro-flora. Although the native micro-flora did not decrease L. monocytogenes to below limits of detection, it was enough of a decrease to warrant further investigation.

High throughput RNAi assay optimization using adherent cell cytometry

PubMed Central

2011-01-01

Background siRNA technology is a promising tool for gene therapy of vascular disease. Due to the multitude of reagents and cell types, RNAi experiment optimization can be time-consuming. In this study adherent cell cytometry was used to rapidly optimize siRNA transfection in human aortic vascular smooth muscle cells (AoSMC). Methods AoSMC were seeded at a density of 3000-8000 cells/well of a 96well plate. 24 hours later AoSMC were transfected with either non-targeting unlabeled siRNA (50 nM), or non-targeting labeled siRNA, siGLO Red (5 or 50 nM) using no transfection reagent, HiPerfect or Lipofectamine RNAiMax. For counting cells, Hoechst nuclei stain or Cell Tracker green were used. For data analysis an adherent cell cytometer, Celigo® was used. Data was normalized to the transfection reagent alone group and expressed as red pixel count/cell. Results After 24 hours, none of the transfection conditions led to cell loss. Red fluorescence counts were normalized to the AoSMC count. RNAiMax was more potent compared to HiPerfect or no transfection reagent at 5 nM siGLO Red (4.12 +/-1.04 vs. 0.70 +/-0.26 vs. 0.15 +/-0.13 red pixel/cell) and 50 nM siGLO Red (6.49 +/-1.81 vs. 2.52 +/-0.67 vs. 0.34 +/-0.19). Fluorescence expression results supported gene knockdown achieved by using MARCKS targeting siRNA in AoSMCs. Conclusion This study underscores that RNAi delivery depends heavily on the choice of delivery method. Adherent cell cytometry can be used as a high throughput-screening tool for the optimization of RNAi assays. This technology can accelerate in vitro cell assays and thus save costs. PMID:21518450

Mathematic modeling the relationship of bacteria number in a dairy product and the color difference measured by a CCD image sensor

NASA Astrophysics Data System (ADS)

Zhou, Zhen; Zhao, Zhigang; Chen, Dongkui; Liu, Yuping

2005-01-01

Although many methods, such as bacteria plate count, flow cytometry and impedance method have been broadly used in the dairy industry to quantitate bacteria numbers around the world, none of them is a quick, low cost and easy one. In this study, we proposed to apply the color difference theory in this field to establish a mathematic model to quantitate bacteria number in fresh milk. Preliminary testing results not only indicate that the application of the color difference theory to the new system is practical, but also confirm the theoretical relationship between the numbers of bacteria, incubation time and color difference. The proof of the principal study in this article further suggests that the novel method has the potential to replace the traditional methods to determine bacteria numbers for the food industry.

Assimilable organic carbon (AOC) determination using GFP-tagged Pseudomonas fluorescens P-17 in water by flow cytometry.

PubMed

Tang, Peng; Wu, Jie; Liu, Hou; Liu, Youcai; Zhou, Xingding

2018-01-01

One of the newly developed methods for Assimilable organic carbon (AOC) determination is leveraged on the cell enumeration by flow cytometry (FC) which could provide a rapid and automated solution for AOC measurement. However, cell samples staining with fluorescence dye is indispensable to reduce background and machine noise. This step would bring additional cost and time consuming for this method. In this study, a green fluorescence protein (GFP) tagged strain derived of AOC testing strain Pseudomonas fluorescens P-17 (GFP-P17) was generated using Tn5 transposon mutagenesis. Continuous culture of this mutant GFP-P17 showed stable expression of eGFP signal detected by flow cytometry without staining step. In addition, this GFP-P17 strain displayed faster growth rate and had a wider range of carbon substrate utilization patterns as compared with P17 wild-type. With this strain, the capability of a new FC method with no dye staining was explored in standard acetate solution, which suggests linear correlation of counts with acetate carbon concentration. Furthermore, this FC method with GFP-P17 strain is applicable in monitoring GAC/BAC efficiency and condition as similar trends of AOC level in water treatment process were measured by both FC method and conventional spread plating count method. Therefore, this fast and easily applicable GFP-P17 based FC method could serve as a tool for routine microbiological drinking water monitoring.

An investigation of the antibacterial ability and cytotoxicity of a novel cu-bearing 317L stainless steel

NASA Astrophysics Data System (ADS)

Sun, Da; Xu, Dake; Yang, Chunguang; Shahzad, M. Babar; Sun, Ziqing; Xia, Jin; Zhao, Jinlong; Gu, Tingyue; Yang, Ke; Wang, Guixue

2016-07-01

In order to solve the challenging problem of microbial infections caused by microorganisms on medical implants, it is imperative to develop novel antimicrobial biomaterials. This work demonstrated that 317L-Cu stainless steel (SS), created by adding copper through a solution and aging heat treatment process, exhibited good antibacterial properties against staphylococcus aureus, achieving 2 log reduction of planktonic cells after 5 days of incubation. In this study, the antibacterial test was performed using the plate count method, the fluorescence cell staining method and the quantitative polymerase chain reaction (qPCR) method. It is well known that a high concentration of copper ion can lead to cytotoxicity. This work explored the cytotoxicity of 317L-Cu SS through real-time cell analysis (RTCA). Experimental results demonstrated that the 317L-Cu SS possessed a satisfactory antibacterial ability against S. aureus, and the antibacterial rate based on the reduction of sessile cell count reached 98.3% after 24-hour treatment. The bacterial adhesion and the biofilm thickness were considerably reduced by the 317L-Cu SS. The results of RTCA suggested that 317L-Cu SS did not introduce cytotoxicity to mouse cells, indicating its suitability as a medical implant material.

Development of PMA real-time PCR method to quantify viable cells of Pantoea agglomerans CPA-2, an antagonist to control the major postharvest diseases on oranges.

PubMed

Soto-Muñoz, Lourdes; Teixidó, Neus; Usall, Josep; Viñas, Inmaculada; Crespo-Sempere, Ana; Torres, Rosario

2014-06-16

Dilution plating is the quantification method commonly used to estimate the population level of postharvest biocontrol agents, but this method does not permit a distinction among introduced and indigenous strains. Recently, molecular techniques based on DNA amplification such as quantitative real-time PCR (qPCR) have been successfully applied for their high strain-specific detection level. However, the ability of qPCR to distinguish viable and nonviable cells is limited. A promising strategy to avoid this issue relies on the use of nucleic acid intercalating dyes, such as propidium monoazide (PMA), as a sample pretreatment prior to the qPCR. The objective of this study was to optimize a protocol based on PMA pre-treatment samples combined with qPCR to distinguish and quantify viable cells of the biocontrol agent P. agglomerans CPA-2 applied as a postharvest treatment on orange. The efficiency of PMA-qPCR method under the established conditions (30μM PMA for 20min of incubation followed by 30min of LED light exposure) was evaluated on an orange matrix. Results showed no difference in CFU or cells counts of viable cells between PMA-qPCR and dilution plating. Samples of orange matrix inoculated with a mixture of viable/dead cells showed 5.59log10 CFU/ml by dilution plating, 8.25log10 cells/ml by qPCR, and 5.93log10 cells/ml by PMA-qPCR. Furthermore, samples inoculated with heat-killed cells were not detected by dilution plating and PMA-qPCR, while by qPCR was of 8.16log10 cells/ml. The difference in quantification cycles (Cq) among qPCR and PMA-qPCR was approximately 16cycles, which means a reduction of 65,536 fold of the dead cells detected. In conclusion, PMA-qPCR method is a suitable tool for quantify viable CPA-2 cells, which could be useful to estimate the ability of this antagonist to colonize the orange surface. Copyright © 2014 Elsevier B.V. All rights reserved.

MEASUREMENT OF THE INTENSITY OF THE PROTON BEAM OF THE HARVARD UNIVERSITY SYNCHROCYCLOTRON FOR ENERGY-SPECTRAL MEASUREMENTS OF NUCLEAR SECONDARIES

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

Santoro, R.T.; Peelle, R.W.

1964-03-01

Two thin helium-filled parallel-plate ionization chambers were designed for use in continuously monitoring the 160-Mev proton beam of the Harvard University Synchrocyclotron over an intensity range from 10/sup 5/ to 10/sup 10/ protons/ sec. The ionlzation chambers were calibrated by two independert methods. In four calibrations the charge collected in the ionization chambers was compared with that deposited in a Faraday cup which followed the ionization chambers in the proton beam. In a second method, a calibration was made by individually counting beam protons with a pnir of thin scintillation detectors. The ionization chamber response was found to be flatmore » within 2% for a five-decade range of beam intensity. Comparison of the Faraday-cup calibrations with that from proton counting shows agreement to within 5%, which is considered satisfactory. The experimental results were also in agreement, within estimated errors, with the ionization chamber response calculated using an accepted value of the average energy loss per ion pair for helium. A slow shift in the calibrations with time is ascribed to a gradual contamination of the helium of the chambers by air leakage. (auth)« less

Spatially explicit models for inference about density in unmarked or partially marked populations

USGS Publications Warehouse

Chandler, Richard B.; Royle, J. Andrew

2013-01-01

Recently developed spatial capture–recapture (SCR) models represent a major advance over traditional capture–recapture (CR) models because they yield explicit estimates of animal density instead of population size within an unknown area. Furthermore, unlike nonspatial CR methods, SCR models account for heterogeneity in capture probability arising from the juxtaposition of animal activity centers and sample locations. Although the utility of SCR methods is gaining recognition, the requirement that all individuals can be uniquely identified excludes their use in many contexts. In this paper, we develop models for situations in which individual recognition is not possible, thereby allowing SCR concepts to be applied in studies of unmarked or partially marked populations. The data required for our model are spatially referenced counts made on one or more sample occasions at a collection of closely spaced sample units such that individuals can be encountered at multiple locations. Our approach includes a spatial point process for the animal activity centers and uses the spatial correlation in counts as information about the number and location of the activity centers. Camera-traps, hair snares, track plates, sound recordings, and even point counts can yield spatially correlated count data, and thus our model is widely applicable. A simulation study demonstrated that while the posterior mean exhibits frequentist bias on the order of 5–10% in small samples, the posterior mode is an accurate point estimator as long as adequate spatial correlation is present. Marking a subset of the population substantially increases posterior precision and is recommended whenever possible. We applied our model to avian point count data collected on an unmarked population of the northern parula (Parula americana) and obtained a density estimate (posterior mode) of 0.38 (95% CI: 0.19–1.64) birds/ha. Our paper challenges sampling and analytical conventions in ecology by demonstrating that neither spatial independence nor individual recognition is needed to estimate population density—rather, spatial dependence can be informative about individual distribution and density.

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

PubMed Central

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

2013-01-01

The purpose of the present study was to determine if the monoculture antifouling effect of several pigmented pseudoalteromonads was retained in in vitro mesocosm systems using natural coastal seawater and when the bacteria were embedded in paint used on surfaces submerged in coastal waters. Pseudoalteromonas piscicida survived on a steel surface and retained antifouling activity for at least 53 days in sterile seawater, whereas P. tunicata survived and had antifouling activity for only 1 week. However, during the first week, all Pseudoalteromonas strains facilitated rather than prevented bacterial attachment when used to coat stainless steel surfaces and submerged in mesocosms with natural seawater. The bacterial density on surfaces coated with sterile growth medium was 105 cells/cm2 after 7 days, whereas counts on surfaces precoated with Pseudoalteromonas were significantly higher, at 106 to 108 cells/cm2. However, after 53 days, seven of eight Pseudoalteromonas strains had reduced total bacterial adhesion compared to the control. P. piscicida, P. antarctica, and P. ulvae remained on the surface, at levels similar to those in the initial coating, whereas P. tunicata could not be detected. Larger fouling organisms were observed on all plates precoated with Pseudoalteromonas; however, plates coated only with sterile growth medium were dominated by a bacterial biofilm. Suspensions of a P. piscicida strain and a P. tunicata strain were incorporated into ship paints (Hempasil x3 87500 and Hempasil 77500) used on plates that were placed at the Hempel A/S test site in Jyllinge Harbor. For the first 4 months, no differences were observed between control plates and treated plates, but after 5 to 6 months, the control plates were more fouled than the plates with pseudoalteromonad-based paint. Our study demonstrates that no single laboratory assay can predict antifouling effects and that a combination of laboratory and real-life methods must be used to determine the potential antifouling capability of new agents or organisms. PMID:23995925

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

PubMed

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

2006-01-01

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

A spore counting method and cell culture model for chlorine disinfection studies of Encephalitozoon syn. Septata intestinalis.

PubMed

Wolk, D M; Johnson, C H; Rice, E W; Marshall, M M; Grahn, K F; Plummer, C B; Sterling, C R

2000-04-01

The microsporidia have recently been recognized as a group of pathogens that have potential for waterborne transmission; however, little is known about the effects of routine disinfection on microsporidian spore viability. In this study, in vitro growth of Encephalitozoon syn. Septata intestinalis, a microsporidium found in the human gut, was used as a model to assess the effect of chlorine on the infectivity and viability of microsporidian spores. Spore inoculum concentrations were determined by using spectrophotometric measurements (percent transmittance at 625 nm) and by traditional hemacytometer counting. To determine quantitative dose-response data for spore infectivity, we optimized a rabbit kidney cell culture system in 24-well plates, which facilitated calculation of a 50% tissue culture infective dose (TCID(50)) and a minimal infective dose (MID) for E. intestinalis. The TCID(50) is a quantitative measure of infectivity and growth and is the number of organisms that must be present to infect 50% of the cell culture wells tested. The MID is as a measure of a system's permissiveness to infection and a measure of spore infectivity. A standardized MID and a standardized TCID(50) have not been reported previously for any microsporidian species. Both types of doses are reported in this paper, and the values were used to evaluate the effects of chlorine disinfection on the in vitro growth of microsporidia. Spores were treated with chlorine at concentrations of 0, 1, 2, 5, and 10 mg/liter. The exposure times ranged from 0 to 80 min at 25 degrees C and pH 7. MID data for E. intestinalis were compared before and after chlorine disinfection. A 3-log reduction (99.9% inhibition) in the E. intestinalis MID was observed at a chlorine concentration of 2 mg/liter after a minimum exposure time of 16 min. The log(10) reduction results based on percent transmittance-derived spore counts were equivalent to the results based on hemacytometer-derived spore counts. Our data suggest that chlorine treatment may be an effective water treatment for E. intestinalis and that spectrophotometric methods may be substituted for labor-intensive hemacytometer methods when spores are counted in laboratory-based chlorine disinfection studies.

Inactivation of Salmonella typhimurium and Escherichia coli O157:H7 in apple juice by a combination of nisin and cinnamon.

PubMed

Yuste, J; Fung, D Y C

2004-02-01

Pasteurized apple juice with nisin (0, 25, 50, 100, and 200 ppm, wt/vol) and cinnamon (0 and 0.3%, wt/vol) was inoculated with Salmonella Typhimurium and Escherichia coli O157:H7 at 10(4) CFU/ml and stored at 5 and 20 degrees C. Counts on tryptic soy agar (TSA), selective medium (xylose Lysine desoxycholate agar for Salmonella Typhimurium, and MacConkey sorbitol agar for E. coli O157:H7), and thin agar layer (TAL) were determined at 1 h and 1, 3, 7, and 14 days. The TAL method (selective medium overlaid with TSA) was used for recovery of sublethally injured cells. The pathogens were gradually inactivated by the acidic pH of apple juice. Nisin and cinnamon greatly contributed to the inactivation. The killing effect was more marked at 20 degrees C, with counts in all treated samples being undetectable by direct plating in 3 days for Salmonella Typhimurium and 7 days for E. coli O157:H7. Thus, several factors influenced the decrease in counts: low pH, addition of nisin and cinnamon, and storage temperature. The TAL method was as effective as TSA in recovering injured cells of the pathogens. The combination of nisin and cinnamon accelerates death of Salmonella Typhimurium and E. coli O157:H7 in apple juice and so enhances the safety of the product.

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

PubMed

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 < 0.05), whereas the other FABs contained , 25 CFU/25 ml microorganisms. Unexpectedly, neither microbial diversity nor microbial count correlated with the alcohol content (4.7 to 14.1%) or pH (3.4 to 4.2) of the product. Despite the harsh conditions, coliforms (detected in 23.8% of microbrewed 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.

Chlorophyll mediated photodynamic inactivation of blue laser on Streptococcus mutans

NASA Astrophysics Data System (ADS)

Astuti, Suryani Dyah; Zaidan, A.; Setiawati, Ernie Maduratna; Suhariningsih

2016-03-01

Photodynamic inactivation is an inactivation method in microbial pathogens that utilize light and photosensitizer. This study was conducted to investigate photodynamic inactivation effects of low intensity laser exposure with various dose energy on Streptococcus mutans bacteria. The photodynamic inactivation was achieved with the addition of chlorophyll as photosensitizers. To determine the survival percentage of Streptococcus mutans bacteria after laser exposure, the total plate count method was used. For this study, the wavelength of the laser is 405 nm and variables of energy doses are 1.44, 2.87, 4.31, 5.74, 7.18, and 8.61 in J/cm2. The results show that exposure to laser with energy dose of 7.18 J/cm2 has the best photodynamic inactivation with a decrease of 78% in Streptococcus

Integral Airframe Structures (IAS): Validated Feasibility Study of Integrally Stiffened Metallic Fuselage Panels for Reducing Manufacturing Costs

NASA Technical Reports Server (NTRS)

Munroe, J.; Wilkins, K.; Gruber, M.; Domack, Marcia S. (Technical Monitor)

2000-01-01

The Integral Airframe Structures (IAS) program investigated the feasibility of using "integrally stiffened" construction for commercial transport fuselage structure. The objective of the program was to demonstrate structural performance and weight equal to current "built-up" structure with lower manufacturing cost. Testing evaluated mechanical properties, structural details, joint performance, repair, static compression, and two-bay crack residual strength panels. Alloys evaluated included 7050-T7451 plate, 7050-T74511 extrusion, 6013-T6511x extrusion, and 7475-T7351 plate. Structural performance was evaluated with a large 7475-T7351 pressure test that included the arrest of a two-bay longitudinal crack, and a measure of residual strength for a two-bay crack centered on a broken frame. Analysis predictions for the two-bay longitudinal crack panel correlated well with the test results. Analysis activity conducted by the IAS team strongly indicates that current analysis tools predict integral structural behavior as accurately as built-up structure. The cost study results indicated that, compared to built-up fabrication methods, high-speed machining structure from aluminum plate would yield a recurring cost savings of 61%. Part count dropped from 78 individual parts on a baseline panel to just 7 parts for machined IAS structure.

40 CFR 141.72 - Disinfection.

Code of Federal Regulations, 2011 CFR

2011-07-01

... serves water to the public. Water in the distribution system with a heterotrophic bacteria concentration... heterotrophic bacteria plate count (HPC) is measured; c=number of instances where the residual disinfectant... system with a heterotrophic bacteria concentration less than or equal to 500/ml, measured as...

40 CFR 141.72 - Disinfection.

Code of Federal Regulations, 2013 CFR

2013-07-01

... serves water to the public. Water in the distribution system with a heterotrophic bacteria concentration... heterotrophic bacteria plate count (HPC) is measured; c=number of instances where the residual disinfectant... system with a heterotrophic bacteria concentration less than or equal to 500/ml, measured as...

High Speed Large Format Photon Counting Microchannel Plate Imaging Sensors

NASA Astrophysics Data System (ADS)

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

The development of a new class of microchannel plate technology, using atomic layer deposition (ALD) techniques applied to a borosilicate microcapillary array is enabling the implementation of larger, more stable detectors for Astronomy and remote sensing. Sealed tubes with MCPs with SuperGenII, bialkali, GaAs and GaN photocathodes have been developed to cover a wide range of optical/UV sensing applications. Formats of 18mm and 25mm circular, and 50mm (Planacon) and 20cm square have been constructed for uses from night time remote reconnaissance and biological single-molecule fluorescence lifetime imaging microscopy, to large area focal plane imagers for Astronomy, neutron detection and ring imaging Cherenkov detection. The large focal plane areas were previously unattainable, but the new developments in construction of ALD microchannel plates allow implementation of formats of 20cm or more. Continuing developments in ALD microchannel plates offer improved overall sealed tube lifetime and gain stability, and furthermore show reduced levels of radiation induced background. High time resolution astronomical and remote sensing applications can be addressed with microchannel plate based imaging, photon time tagging detector sealed tube schemes. Photon counting imaging readouts for these devices vary from cross strip (XS), cross delay line (XDL), to stripline anodes, and pad arrays depending on the intended application. The XS and XDL readouts have been implemented in formats from 22mm, and 50mm to 20cm. Both use MCP charge signals detected on two orthogonal layers of conductive fingers to encode event X-Y positions. XDL readout uses signal propagation delay to encode positions while XS readout uses charge cloud centroiding. Spatial resolution readout of XS detectors can be better than 20 microns FWHM, with good image linearity while using low gain (<10^6), allowing high local counting rates and longer overall tube lifetime. XS tubes with electronics can encode event rates of >5 MHz and event timing accuracy of ~100ps. We will discuss how we are applying these detector system developments for devices in formats of 18mm and 25mm circular, and 50mm and 20cm square. The performance characteristics will be demonstrated along with lifetest data taken over the last year. Implications for ground based instruments to study transient and variable astronomical objects, as well as implementation in satellite instruments for earth atmospheric, planetary and solar observations will be discussed.

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

PubMed

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

2017-07-01

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

Liming Poultry Manures to Kill Pathogens and Decrease Soluble Phosphorus

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

Maguire,R.; Hesterberg, D.; Gernat, A.

2006-01-01

Received for publication September 9, 2005. Stabilizing phosphorus (P) in poultry waste to reduce P losses from manured soils is important to protect surface waters, while pathogens in manures are an emerging issue. This study was conducted to evaluate CaO and Ca(OH){sub 2} for killing manure bacterial populations (pathogens) and stabilizing P in poultry wastes and to investigate the influence on soils following amendment with the treated wastes. Layer manure and broiler litter varying in moisture content were treated with CaO and Ca(OH){sub 2} at rates of 2.5, 5, 10, and 15% by weight. All treated wastes were analyzed formore » microbial plate counts, pH, and water-soluble phosphorus (WSP), while a few selected layer manures were analyzed by phosphorus X-ray absorption near edge structure (XANES). A loamy sand and a silt loam were amended with broiler litter and layer manure treated with CaO at rates of 0, 2.5, 5, 10, and 15% and soil WSP and pH were measured at times 1, 8, and 29 d. Liming reduced bacterial populations, with greater rates of lime leading to greater reductions; for example 10% CaO applied to 20% solids broiler litter reduced the plate counts from 793 000 to 6500 mL{sup -1}. Liming also reduced the WSP in the manures by over 90% in all cases where at least 10% CaO was added. Liming the manures also reduced WSP in soils immediately following application and raised soil pH. The liming process used successfully reduced plate counts and concerns about P losses in runoff following land application of these limed products due to decreased WSP.« less

Microbial quality and associated health risks of raw milk marketed in the Tanga region of Tanzania.

PubMed

Swai, E S; Schoonman, L

2011-06-01

To evaluate microbial quality and associated health risks of raw milk marketed in the Tanga region of Tanzania. A microbial quality assessment of marketed raw milk was undertaken by evaluating 59 samples of milk from selling points (collecting centres =15), bicycle boys (12) and kiosks/restaurants (32) in Tanga city during April-May 2005. Quality and milk-borne hazards were assessed using a combination of tests in order to quantify the occurrence of Brucellosis (milk ring test), Escherichia coli (E. coli) O157:H7 (culture), the coliform bacteria as well as standard plate count (SPC). Specific gravity (SG) determination was used as an indicator of adulteration. The mean coliform plate count (c.f.u/mL) of milk handled by bicycle boys (4.2×10(6)) was significantly higher than that handled by collecting centres (3.0×10(6)) and kiosk/ restaurants (1.4×10(6)), respectively (P < 0.05). Of the 59 milk samples collected, 33 (56%) were Brucella milk ring test (MRT)-positive and 78% and 17% of the samples graded satisfactorily based on SG and coliform plate counts as prescribed by East African Community standards for raw milk. There was no verocytotoxigenic E. coli (VTEC) O157: H7 in any of the milk samples collected and analysed during the present study. It can be concluded that raw market milk in the study area is of poor bacteriological quality and hazardous for human consumption. This highlights the need to implement good hygiene practices and effective monitoring from production through the delivery chain to the consumer. Further studies are needed for detection of toxins that are produced by E. coli, other pathogenic spore forming bacteria (Bacillus spp. and Clostridium spp.) and other harmful microorganisms.

Liming poultry manures to decrease soluble phosphorus and suppress the bacteria population.

PubMed

Maguire, R O; Hesterberg, D; Gernat, A; Anderson, K; Wineland, M; Grimes, J

2006-01-01

Stabilizing phosphorus (P) in poultry waste to reduce P losses from manured soils is important to protect surface waters, while pathogens in manures are an emerging issue. This study was conducted to evaluate CaO and Ca(OH)2 for killing manure bacterial populations (pathogens) and stabilizing P in poultry wastes and to investigate the influence on soils following amendment with the treated wastes. Layer manure and broiler litter varying in moisture content were treated with CaO and Ca(OH)2 at rates of 2.5, 5, 10, and 15% by weight. All treated wastes were analyzed for microbial plate counts, pH, and water-soluble phosphorus (WSP), while a few selected layer manures were analyzed by phosphorus X-ray absorption near edge structure (XANES). A loamy sand and a silt loam were amended with broiler litter and layer manure treated with CaO at rates of 0, 2.5, 5, 10, and 15% and soil WSP and pH were measured at times 1, 8, and 29 d. Liming reduced bacterial populations, with greater rates of lime leading to greater reductions; for example 10% CaO applied to 20% solids broiler litter reduced the plate counts from 793,000 to 6500 mL-1. Liming also reduced the WSP in the manures by over 90% in all cases where at least 10% CaO was added. Liming the manures also reduced WSP in soils immediately following application and raised soil pH. The liming process used successfully reduced plate counts and concerns about P losses in runoff following land application of these limed products due to decreased WSP.

Effects of 1-MeV gamma radiation on a multi-anode microchannel array detector tube

NASA Technical Reports Server (NTRS)

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

1979-01-01

A multianode microchannel array (MAMA) detector tube without a photocathode was exposed to a total dose of 1,000,000 rads of 1-MeV gamma radiation from a Co-60 source. The high-voltage characteristic of the microchannel array plate, average dark count, gain, and resolution of pulse height distribution characteristics showed no degradation after this total dose. In fact, the degassing of the microchannels induced by the high radiation flux had the effect of cleaning up the array plate and improving its characteristics.

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

PubMed

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

2012-08-01

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

Further studies on Egyptian soil fungi: succession of sugar and osmophilic fungi in soil amended with five organic substrates.

PubMed

Shaban, G M

1996-01-01

The sugar and osmophilic fungal composition of soils amended with five organic substrates (newspaper, orange peel, bromegrass leaves, wheat straw and wood sawdust) was estimated after 2, 4, 6, 8 and 10 weeks using the dilution plate method on glucose and 50% sucrose Czapek's agar media. Wheat straw was the best substrate for total counts of both sugar and osmophilic fungi followed by newspaper, bromegrass leaves, wood sawdust and orange peel. Wood sawdust supported the highest average counts of total sugar fungi, Fusarium, Mucor, Scopulariopsis, Trichoderma and Trimmatostroma spp.; Newspaper, of Aspergillus (8 spp.), Penicillium (4 spp.) and Chaetomium sp.; bromegrass leaves of Cladosporium sp., Humicola sp. and Sporotrichum sp.; orange peel, of Alternaria sp., Circinella sp. and Stachybotrys sp.; and wheat straw, of Botryotrichum sp. and Myrothecium sp. Bromegrass leaves and orange peel supported the highest average counts of total osmophilic fungi, Aspergillus (10 spp.), Cladosporium sp. Paecillomyces sp. and Rhizopus sp.; and of Stemphylium sp., Trichoderma sp., Humicola sp. and Circinella sp. respectively; wheat straw, of Epicoccum sp., Scopulariopsis sp. and Trichothecium sp.; newspaper, of Penicillium (4 spp.) and Alternaria sp.; and wood sawdust of Curvularia sp. and Fusarium (3 spp.). The best colonizers throughout the experimental periods were Aspergilus and Penicillium spp.

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

PubMed

Yasuda, Naruomi; Suzuki, Hitoshi; Katafuchi, Tetsuro

2016-01-01

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

Microbiological baseline study of poultry slaughtered in provincially inspected abattoirs in Alberta, Canada

PubMed Central

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

2009-01-01

Studies to determine baseline levels of microbial contaminants and foodborne bacterial pathogens are needed to evaluate the effectiveness of Hazard Analysis Critical Control Point (HACCP) programs, Good Manufacturing/Production Practices, and various interventions. In 2004 and 2005 poultry carcass rinses from provincially inspected abattoirs in Alberta, Canada, were tested to determine the levels of aerobic plate count bacteria, coliform bacteria, and generic Escherichia coli, the prevalence and levels of Campylobacter spp., and the prevalence of Salmonella spp. and Shiga toxin-producing E. coli (STEC). Samples were collected from 3 high volume and 62 low volume abbatoirs. All samples (1296) were positive for aerobic plate count bacteria, with 98.8% of samples having counts of 100 000 or less colony forming units (CFU)/cm2. Coliform bacteria were isolated from 99.7% of the 1296 carcasses and were recovered at levels of ≤ 1000 CFU/cm2 for 98.3% of the samples. Generic E. coli were recovered from 99.1% of the 1296 carcasses at levels of ≤ 1000 CFU/cm2 for 98.6% of the samples. Seventy five percent of 1234 samples that were tested for Campylobacter were positive; 37.5% of 1295 samples that were tested for Salmonella were positive; and only 2 of 1296 samples tested for STEC were positive (0.15%). PMID:19412397

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

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

Menlove, Howard Olsen; Henzlova, Daniela

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

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

PubMed

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

2012-06-06

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

Microbiological quality of frozen cauliflower, corn, and peas obtained at retail markets.

PubMed Central

Barnard, R J; Duran, A P; Swartzentruber, A; Schwab, A H; Wentz, B A; Read, R B

1982-01-01

The microbiological quality of blanched frozen cauliflower, cut corn, and peas at the retail level was determined. At 35 degrees C, mean aerobic plate count (APC) values for cauliflower, corn, and peas, respectively, were 30,000, 6,100, and 4,700 per g; at 30 degrees C, the mean APC values were 45,000, 8,500, and 6,800 per g, respectively. Geometric means for coliform, Escherichia coli, and Staphylococcus aureus counts for all three vegetables were less than 10 per g. PMID:6751226

Microbiological survey of five poultry processing plants in the UK.

PubMed

Mead, G C; Hudson, W R; Hinton, M H

1993-07-01

1. Neck skin samples were taken from chickens and turkeys at all the main stages of processing to monitor changes in total viable count (TVC) and counts of coliforms and pseudomonads. 2. Processing reduced TVC by up to 100-fold. Geometric mean counts after packaging were log10 4.4 to 5.3 CFU/g whilst corresponding counts of coliforms were 2.7 to 3.8 CFU/g. 3. Increases in mean TVC or coliforms as a result of either defeathering or evisceration did not exceed 0.6 log. 4. Pseudomonads represented only a minor fraction of the initial microflora of the bird and were often reduced by scalding to a figure which could not be detected by direct plating of samples; however, subsequent contamination resulted in means between log10 2.9 and 4.0 CFU/g for packaged carcases. 5. Although Staphylococcus aureus was readily isolated from defeathering equipment, mean counts from defeathered carcases were always below log10 3.0 CFU/g.

Microbial quality of catfish nuggets

USDA-ARS?s Scientific Manuscript database

The microbiological quality of catfish nuggets is not known. Nuggets, purchased from local retailers in the northeast United States (NJ, NY, PA, and DE), were tested for aerobic plate count (APC) at 22 and 37 deg C, Enterobacteriacea, and Escherichia coli/coliform using Petrifilms**™. The BAX**™ ...

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

PubMed

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.

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

PubMed

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

1980-06-01

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

A study of psychrophilic organisms isolated from the manufacture and assembly areas of spacecraft to be used in the Viking mission

NASA Technical Reports Server (NTRS)

Foster, T. L.

1974-01-01

The effect of storage of dry heat treated Teflon ribbons under nitrogen gas followed by high vacuum on the recovery of hardy organisms from the ribbons was studied. A similar experiment was performed on spore crops of hardy organisms recovered previously from Cape Canaveral. Hardy organisms have been inoculated onto slides and subjected to an artificial Martian environment in an attempt to demonstrate their growth in this environment. Additional experiments using the artificial Martian environment include response of soil samples from the VAB with both constant temperature and freeze-thaw cycles. These experiments were performed with dried soil and soil containing added water. Other investigations included the effect of heatshock on soil samples, psychrophilic counts of new soil samples from the manufacture area of the Viking spacecraft, effect of pour plate versus spread plate on psychrophilic counts, and preparation of spore crops of hardy organisms from Cape Canaveral.

Electrochemical biosensors for Salmonella: State of the art and challenges in food safety assessment.

PubMed

Silva, Nádia F D; Magalhães, Júlia M C S; Freire, Cristina; Delerue-Matos, Cristina

2018-01-15

According to the recent statistics, Salmonella is still an important public health issue in the whole world. Legislated reference methods, based on counting plate methods, are sensitive enough but are inadequate as an effective emergency response tool, and are far from a rapid device, simple to use out of lab. An overview of the commercially available rapid methods for Salmonella detection is provided along with a critical discussion of their limitations, benefits and potential use in a real context. The distinguished potentialities of electrochemical biosensors for the development of rapid devices are highlighted. The state-of-art and the newest technologic approaches in electrochemical biosensors for Salmonella detection are presented and a critical analysis of the literature is made in an attempt to identify the current challenges towards a complete solution for Salmonella detection in microbial food control based on electrochemical biosensors. Copyright © 2017 Elsevier B.V. All rights reserved.

Integration of Pseudomonas aeruginosa and Legionella pneumophila in drinking water biofilms grown on domestic plumbing materials.

PubMed

Moritz, Miriam M; Flemming, Hans-Curt; Wingender, Jost

2010-06-01

Drinking water biofilms were grown on coupons of plumbing materials, including ethylene-propylene-diene-monomer (EPDM) rubber, silane cross-linked polyethylene (PE-X b), electron-ray cross-linked PE (PE-X c) and copper under constant flow-through of cold tap water. After 14 days, the biofilms were spiked with Pseudomonas aeruginosa, Legionella pneumophila and Enterobacter nimipressuralis (10(6) cells/mL each). The test bacteria were environmental isolates from contamination events in drinking water systems. After static incubation for 24 h, water flow was resumed and continued for 4 weeks. Total cell count and heterotrophic plate count (HPC) of biofilms were monitored, and P. aeruginosa, L. pneumophila and E. nimipressuralis were quantified, using standard culture-based methods or culture-independent fluorescence in situ hybridization (FISH). After 14 days total cell counts and HPC values were highest on EPDM followed by the plastic materials and copper. P. aeruginosa and L. pneumophila became incorporated into drinking water biofilms and were capable to persist in biofilms on EPDM and PE-X materials for several weeks, while copper biofilms were colonized only by L. pneumophila in low culturable numbers. E. nimipressuralis was not detected in any of the biofilms. Application of the FISH method often yielded orders of magnitude higher levels of P. aeruginosa and L. pneumophila than culture methods. These observations indicate that drinking water biofilms grown under cold water conditions on domestic plumbing materials, especially EPDM and PE-X in the present study, can be a reservoir for P. aeruginosa and L. pneumophila that persist in these habitats mostly in a viable but non-culturable state.

Persistence and Viability of Lecanicillium lecanii in Chinese Agricultural Soil

PubMed Central

Peng, De-Liang; Zhou, Jie; Zhang, Xiao-Lin; Zhang, Zhao-Rong; Zhao, Jin-Jin; Wu, Yu-Huan

2015-01-01

The entomopathogenic fungus L. lecanii has been developed as biopesticides and used widely for biological control of several insects in agricultural practice. Due to the lack of isolation/count methods for L. lecanii in soil, the persistence of this fungus in soil appears to have attracted no attention. A selective medium and count method for L. lecanii in soil based on cetyl trimethyl ammonium bromide (CTAB) was developed, and then the persistence and viability of this fungus in soil were investigated under field conditions between 2012 and 2014. The results showed that the rate of recovery for L. lecanii in soil on the selective CTAB medium was satisfactory. The minimum CFUs for L. lecanii on the selective medium (0.5 g/L CTAB) was about 102 conidia/g soil. The L. lecanii density in soil declined quickly in the first month after inoculation with fungal conidia, kept stable for 6 to 10 months, and then decreased gradually until undetectable. L. lecanii could persist for at least 14 months in the agricultural soil of northern China. The colony growth, conidia yield and germination rate on plates, as well as the median lethal concentration or times (LC50 or LT50) to aphids, mycelium growth in aphids and sporulation on aphids of L. lecanii did not change significantly during the persistence in soil. In general, the count method developed here was a very useful tool for monitoring the dynamics of natural or introduced L. lecanii populations in soil, and the data on the persistence of L. lecanii in soil reported here were helpful for biological control and environmental risk assessment. PMID:26375030

Exploring the potential environmental functions of viable but non-culturable bacteria.

PubMed

Su, Xiaomei; Chen, Xi; Hu, Jinxing; Shen, Chaofeng; Ding, Linxian

2013-12-01

A conventional plate count is the most commonly employed method to estimate the number of living bacteria in environmental samples. In fact, judging the level of viable culture by plate count is limited, because it is often several orders of magnitude less than the number of living bacteria actually present. Most of the bacteria are in "viable but non-culturable" (VBNC) state, whose cells are intact and alive and can resuscitate when surrounding conditions are more favorable. The most exciting recent development in resuscitating VBNC bacteria is a bacterial cytokine, namely, the resuscitation-promoting factor (Rpf), secreted by Micrococcus luteus, which promotes the resuscitation and growth of high G+C Gram-positive organisms, including some species of the genus Mycobacterium. However, most of studies deal with VBNC bacteria only from the point of view of medicine and epidemiology. It is therefore of great significance to research whether these VBNC state bacteria also possess some useful environmental capabilities, such as degradation, flocculation, etc. Further studies are needed to elucidate the possible environmental role of the VBNC bacteria, rather than only considering their role as potential pathogens from the point view of epidemiology and public health. We have studied the resuscitation of these VBNC bacteria in polluted environments by adding culture supernatant containing Rpf from M. luteus, and it was found that, as a huge microbial resource, VBNC bacteria could provide important answers to dealing with existing problems of environmental pollution. This mini-review will provide new insight for considering the potentially environmental functions of VBNC bacteria.

Spatial arrangement and size distribution of normal faults, Buckskin detachment upper plate, Western Arizona

NASA Astrophysics Data System (ADS)

Laubach, S. E.; Hundley, T. H.; Hooker, J. N.; Marrett, R. A.

2018-03-01

Fault arrays typically include a wide range of fault sizes and those faults may be randomly located, clustered together, or regularly or periodically located in a rock volume. Here, we investigate size distribution and spatial arrangement of normal faults using rigorous size-scaling methods and normalized correlation count (NCC). Outcrop data from Miocene sedimentary rocks in the immediate upper plate of the regional Buckskin detachment-low angle normal-fault, have differing patterns of spatial arrangement as a function of displacement (offset). Using lower size-thresholds of 1, 0.1, 0.01, and 0.001 m, displacements range over 5 orders of magnitude and have power-law frequency distributions spanning ∼ four orders of magnitude from less than 0.001 m to more than 100 m, with exponents of -0.6 and -0.9. The largest faults with >1 m displacement have a shallower size-distribution slope and regular spacing of about 20 m. In contrast, smaller faults have steep size-distribution slopes and irregular spacing, with NCC plateau patterns indicating imposed clustering. Cluster widths are 15 m for the 0.1-m threshold, 14 m for 0.01-m, and 1 m for 0.001-m displacement threshold faults. Results demonstrate normalized correlation count effectively characterizes the spatial arrangement patterns of these faults. Our example from a high-strain fault pattern above a detachment is compatible with size and spatial organization that was influenced primarily by boundary conditions such as fault shape, mechanical unit thickness and internal stratigraphy on a range of scales rather than purely by interaction among faults during their propagation.

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

PubMed

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

2014-07-01

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

PATHOGENICITY OF DRINKING WATER ISOLATES OF HETEROTROPHIC BACTERIA WITH PUTATIVE VIRULENCE FACTORS

EPA Science Inventory

Although the heterotrophic plate count (HPC) bacteria normally found in potable water are not a threat to the healthy population, some of them may be opportunistic pathogens that could cause adverse health effects in individuals with impaired immune systems. Earlier studies of t...

The development of methods for the detection of Salmonella in chickens by a combination of immunomagnetic separation and PCRs.

PubMed

Dai, Fengying; Zhang, Miao; Xu, Dixin; Yang, Yin; Wang, Jiaxiao; Li, Mingzhen; Du, Meihong

2017-11-01

Micro- and nanoimmunomagnetic beads (MIMBs and NIMBs) used for immunomagnetic separation (IMS) with PCR were studied for the rapid detection of Salmonella. The capture efficiency of the two different IMBs was evaluated by a conventional plate counting method, and the binding pattern was studied using scanning electron microscopy. The specificity of the IMBs was tested with Salmonella, Shigella flexneri, enterohemorrhagic Escherichia coli O157:H7, and Listeria monocytogenes. By comparing the pre-enrichment IMS and the IMS enrichment steps with a 5.5-H enrichment time, this study developed a rapid and sensitive method for the detection of Salmonella in chicken. The method was implemented by IMS enrichment and PCR with MIMBs and NIMBs, with a total analysis time of 8 H. We showed that the method was sensitive based on NIMBs with a detection limit of 10° CFU for Salmonella in 25 g of chicken. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

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

NASA Technical Reports Server (NTRS)

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

2002-01-01

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

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

PubMed

Borneman, Darand L; Ingham, Steve

2014-05-01

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

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

PubMed

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.

Effect of Inoculation Techniques and Relative Humidity on the Growth of Molds on the Surfaces of Yellow Layer Cakes

PubMed Central

Fustier, Patrick; Lafond, Alain; Champagne, Claude P.; Lamarche, François

1998-01-01

Four inoculation techniques were compared for initiation of growth on cake surfaces: spot, air cabinet, spray (atomizer), and talc addition methods. Molds were isolated from commercial cakes and were identified as Aspergillus sydowii, Aspergillus ochraceus, Penicillium funiculosum, and Eurotium herbariorum. Cake surfaces were inoculated with mold spores and incubated under three equilibrium relative humidity (ERH) levels: 97, 85, and 75%. Random contamination by spores in a ventilated air cabinet was the simplest method of inoculation, but standard deviations in the inoculation rates (20% on a relative scale) were almost twice those observed with the other methods. The spot method was the most reproducible. Cake samples inoculated in the air cabinet had colony counts 10 times lower than those obtained for potato dextrose agar plates at 97% ERH, which was not the case with the spray and talc methods. Growth of molds was much slower in the samples incubated in 75% relative humidity, with all methods. Colony counts were generally similar in systems adjusted at 85 to 97% ERH but were lower for samples incubated at 75% ERH. In comparisons of the shelf life estimates obtained by the various inoculation methods, a correlation coefficient (r2) of 0.70 was obtained between the spot method and the other methods of inoculation, while talc, air cabinet, and spray shelf life data were correlated better (r2 ≈ 0.97). The spot method appeared to be the method of choice in consideration of ease of use, precision, and the ability to enable the study of the effects of the environment on mold-free shelf life as well as on the rate of growth of molds on cakes. PMID:16349479

Experimental Design for a Macrofoam Swab Study Relating the Recovery Efficiency and False Negative Rate to Low Concentrations of Two Bacillus anthracis Surrogates on Four Surface Materials

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

Piepel, Gregory F.; Hutchison, Janine R.

2014-04-16

This report describes the experimental design for a laboratory study to quantify the recovery efficiencies and false negative rates of a validated, macrofoam swab sampling method for low concentrations of Bacillus anthracis Sterne (BAS) and Bacillus atrophaeus (BG) spores on four surface materials (stainless steel, glass, vinyl tile, plastic light cover panel). Two analytical methods (plating/counting and polymerase chain reaction) will be used. Only one previous study has investigated false negative as a function of affecting test factors. The surrogates BAS and BG have not been tested together in the same study previously. Hence, this study will provide for completingmore » gaps in the available information on the performance of macrofoam swab sampling at low concentrations.« less

Direct quantification of test bacteria in synthetic water-polluted samples by square wave voltammetry and chemometric methods.

PubMed

Carpani, Irene; Conti, Paolo; Lanteri, Silvia; Legnani, Pier Paolo; Leoni, Erica; Tonelli, Domenica

2008-02-28

A home-made microelectrode array, based on reticulated vitreous carbon, was used as working electrode in square wave voltammetry experiments to quantify the bacterial load of Escherichia coli ATCC 13706 and Pseudomonas aeruginosa ATCC 27853, chosen as test microorganisms, in synthetic samples similar to drinking water (phosphate buffer). Raw electrochemical signals were analysed with partial least squares regression coupled to variable selection in order to correlate these values with the bacterial load estimated by aerobic plate counting. The results demonstrated the ability of the method to detect even low loads of microorganisms in synthetic water samples. In particular, the model detects the bacterial load in the range 3-2,020 CFU ml(-1) for E. coli and in the range 76-155,556 CFU ml(-1) for P. aeruginosa.

Experimental Study and Fractal Analysis on the Anisotropic Performance of Explosively Welded Interfaces of 304 Stainless Steel/245 Carbon Steel

NASA Astrophysics Data System (ADS)

Fu, Yanshu; Qiu, Yaohui; Li, Yulong

2018-03-01

The mechanical anisotropy of an explosive welding composite plate made of 304 stainless steel/245 steel was studied through shear experiments performed on explosively welded wavy interfaces along several orientation angles. The results indicated that the strength and the fracture energy of samples significantly varied with the orientation angles. The fracture surfaces of all samples were observed using a scanning electron microscope and through three-dimensional structure microscopy. The periodic features of all the fracture surfaces were clearly shown in different fracture modes. The fractal dimension of the fracture surfaces was calculated based on the fractal geometry by the box-counting method in MATLAB. The cohesive element model was used to analyze the fracture energy according to the physical dependence of the fractal dimension on thermodynamic entropy and interface separation energy. The fracture energy was an exponential function of the fractal dimension value, which was in good agreement with the experimental results. All results were validated for effective use in the application of anisotropy analysis to the welded interface and structural optimization of explosively welded composite plates.

Experimental Study and Fractal Analysis on the Anisotropic Performance of Explosively Welded Interfaces of 304 Stainless Steel/245 Carbon Steel

NASA Astrophysics Data System (ADS)

Fu, Yanshu; Qiu, Yaohui; Li, Yulong

2018-05-01

The mechanical anisotropy of an explosive welding composite plate made of 304 stainless steel/245 steel was studied through shear experiments performed on explosively welded wavy interfaces along several orientation angles. The results indicated that the strength and the fracture energy of samples significantly varied with the orientation angles. The fracture surfaces of all samples were observed using a scanning electron microscope and through three-dimensional structure microscopy. The periodic features of all the fracture surfaces were clearly shown in different fracture modes. The fractal dimension of the fracture surfaces was calculated based on the fractal geometry by the box-counting method in MATLAB. The cohesive element model was used to analyze the fracture energy according to the physical dependence of the fractal dimension on thermodynamic entropy and interface separation energy. The fracture energy was an exponential function of the fractal dimension value, which was in good agreement with the experimental results. All results were validated for effective use in the application of anisotropy analysis to the welded interface and structural optimization of explosively welded composite plates.

Sensitive far uv spectrograph with a multispectral element microchannel plate detector for rocket-borne astronomy.

PubMed

Weiser, H; Vitz, R C; Moos, H W; Weinstein, A

1976-12-01

An evacuated high transmission prism spectrograph using a microchannel plate detection system with resistive strip readout was flown behind a precision pointing telescope on a sounding rocket. The construction, preparation, flight performance, and calibration stability of the system are discussed. Despite the adverse environmental conditions associated with sounding rocket flights, the microchannel detector system performed well. Far uv spectra (1160-1750 A) of stellar and planetary objects were obtained; spectral features with fluxes as low as 0.06 photons cm(-2) sec(-1) were detectable. This was achieved by operating the plates at lower than normal gains, using sensitive pulse counting electronics with both upper and lower limit discriminators, and maintaining the spectrograph and detector at a pressure of ~10(-6) Torr until reaching altitude.

COMPARISON OF MICROBIAL TRANSFORMATION RATE COEFFICIENTS OF XENOBIOTIC CHEMICALS BETWEEN FIELD-COLLECTED AND LABORATORY MICROCOSM MICROBIOTA

EPA Science Inventory

Two second-order transformation rate coefficients--kb, based on total plate counts, and kA, based on periphyton-colonized surface areas--were used to compare xenobiotic chemical transformation by laboratory-developed (microcosm) and by field-collected microbiota. Similarity of tr...

Bacterial Cleanability of Various Types of Eating Surfaces.

ERIC Educational Resources Information Center

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

1953-01-01

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

[STUDY OF LIPIDS SEED'S OIL OF VITEX AGNUS CASTUS GROWING IN GEORGIA].

PubMed

Kikalishvili, B; Zurabashvili, D; Sulakvelidze, Ts; Malania, M; Turabelidze, D

2016-07-01

There was established the lipid composition of the seeds of Vitex agnus castus L. by the qualitative and quantitative methods of analyses. There were received neutral lipids from the seeds by extraction with hexane in the yield 10%, counted on dry material. For the divide of neutral lipids there was used silica gel plates LS 5/40 in the systems of solvents: 1. petroleum ether-diethylether-acidum aceticum (85:14:1), 2. hexane-diethylether (1:1). After obtaining neutral lipids from the residual plant shrot pollar lipids was extracted with the mixture of chloroform-methanol (2:1) and was divided on silica gel plates LS 5/40, mobile phase: 1. chloroform-methanol-25% ammonium hydrate 2. chloroform-methanol icy acetic acid-water (170:25:25:6). In the sum of polar lipids qualitatively were established phospholipids: lisophosphatidylcholine, phosphatidylinosit, phospatidylethanolamine and N-acylphosphatidylethanolamine, in neutral lipids, hydrocarbons, triglycerids, free fatty acids and sterines. By the method of high performance liquid chromatography analyses there were identified following free fatty acids: lauric, myristic, palmitic, stearic, linolic, linolenic, arachidic and begenic, unsaturated oleic and polyunsaturated linolic and linolenic acids. obtained oil with unique composition from the seeds of Vitex agnus-castus indicates to its high biological activity and importance for usage in medicine.

Suppression of Listeria monocytogenes by the Native Micro-Flora in Teewurst Sausage

PubMed Central

Austin-Watson, Clytrice; Grant, Ar’Quette; Brice, Michline

2013-01-01

Modern consumers are interested in the use of non-chemical methods to control pathogens when heat sterilization is not an option. Such is the case with teewurst sausage, a raw spreadable sausage and a popular German commodity. Although Listeria was not found in teewurst, the optimal microbial growing conditions of teewurst coupled with the ubiquity of L. monocytogenes in nature, makes the possibility of contamination of products very possible. This pilot study was conducted to examine teewurst’s native micro-flora’s ability to suppress the outgrowth of L. monocytogenes at 10 °C using standard plate counts and PCR-DGGE. Traditional plating methods showed L. monocytogenes growth significantly decreased when in competition with the teewurst’s native micro-flora (p < 0.05). The native micro-flora of the teewurst suppressed the overall growth of L. monocytogenes by an average of two logs, under these conditions. Denaturing Gradient Gel Electrophoresis (DGGE) amplicons with unique banding patterns were extracted from DGGE gel for identification. Brochothrix thermosphacta and Lactobacillus curvatus were identified as a part of the teewurst’s native micro-flora. Although the native micro-flora did not decrease L. monocytogenes to below limits of detection, it was enough of a decrease to warrant further investigation. PMID:28239131

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

PubMed

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

2006-09-01

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

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

PubMed

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 old when tested. This may be one of the major factors contributing to the larger goat milk BsnFC equivalence. Correlations and interactions between various test results were also discussed to further understand differences between the 2 methods for goat and cow milks. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Impact of ultraviolet radiation treatments on the quality of freshly prepared tomato (Solanum lycopersicum) juice.

PubMed

Bhat, Rajeev

2016-12-15

Impact of ultraviolet (UV-C) radiation treatments (0, 15, 30 and 60min) on freshly extracted tomato juice quality (physicochemical properties, antioxidant activity and microbial load) was evaluated. On exposure to UV-C, level of water activity, total soluble solids, and titratable acidity exhibited non-significant increase up to 30min of exposure time. Regarding colour analysis, L∗ value significantly increased with subsequent decrease in a∗ and b∗ values post UV-C treatments. Clarity, DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity and total phenolics content significantly increased, whereas ascorbic acid level significantly reduced at 60min of UV-C exposure time. So also, lycopene content exhibited a non-significant decrease after UV-C treatment. Microbial studies showed reduction in total plate count and total mould counts post UV-C treatment. Overall, UV-C treatment being a physical, non-thermal method of food preservation holds the ability to improve or preserve vital quality parameters in freshly prepared tomato juices, and henceforth possesses high scope for commercial exploration. Copyright © 2016 Elsevier Ltd. All rights reserved.

Comparative study on the microbiological features of angular cheilitis in HIV seropositive and HIV seronegative patients from South India

PubMed Central

Krishnan, P Anitha; Kannan, Ranganathan

2013-01-01

Objective: This study was designed to compare the microbiological features of angular cheilitis (AC) in human immunodeficiency virus (HIV) seropositive and HIV seronegative individuals, in a group of south Indians. Materials and Methods: Swabs from oral commissures of 46 patients were obtained and inoculated on to Sabouraud's dextrose agar (SDA) supplemented with chloramphenicol, blood agar (BA) and MacConkey's agar (MCA) plates and cultured. α-hemolytic Streptococci, Staphylococcus albus, Staphylococcus aureus, Candida species, Klebsiella species and Pseudomonas species were cultured. Candidal colonies were further speciated by the conventional biotyping technique. Results: In AC of HIV seropositive patients Candida albicans and Staphylococcus aureus were more prevalent than that in HIV seronegative patients. Incidentally in patients with CD4 cell count less than 200 there was an increase in the incidence of Candidal and Staphylococcus aureus colonization when compared to patients with CD4 cell count higher than 200. Conclusion: The present study suggests a definite difference in the microbial flora of AC in HIV seropositive patients than that of HIV seronegative population. PMID:24574650

[Chromosome analysis of bulls in relation to disorders of sexual activity].

PubMed

Staník, J; Izariková, A

1984-05-01

Chromosomal analysis was used for the examination of 16 bulls of different breeds from the Milhostov breeding station. The examined bulls exhibited disorders of sexual activity (disorders of spermiogenesis, aspermia, bad quality of semen, hypoplasia of testes, etc.). The examination was performed by the method after Moorhead et al. (1960) modified by Lojda et al. (1974): metaphase plates were evaluated microscopically (100 X 12) and from photos. The chromosomes were counted by means of the counting documator (from film negatives) and from photos. A card was prepared for each animal. Hyposomy (11 sires--68.75%) and hyperploidy (10 sires--62.5%) were found to be the most frequent numerical aberrations, followed by polysomy (4 sires--25.0%) and other aneuploidies (one case--6.2%). As to structural defects, breaks occurred in 14 sires (87.5%), bichromatid breaks in five sires (31.25%) and breaks on sexual chromosomes in three sires (18.75%). Centric fusion was observed in one case (6.25%), association in two cases (12.5%) and mixed aberrations in four cases (25.00%).

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

PubMed

MacDonald, David S; Waterfield, J Douglas

2011-01-01

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

Energy resolution of the CdTe-XPAD detector: calibration and potential for Laue diffraction measurements on protein crystals.

PubMed

Medjoubi, Kadda; Thompson, Andrew; Bérar, Jean-François; Clemens, Jean-Claude; Delpierre, Pierre; Da Silva, Paulo; Dinkespiler, Bernard; Fourme, Roger; Gourhant, Patrick; Guimaraes, Beatriz; Hustache, Stéphanie; Idir, Mourad; Itié, Jean-Paul; Legrand, Pierre; Menneglier, Claude; Mercere, Pascal; Picca, Frederic; Samama, Jean-Pierre

2012-05-01

The XPAD3S-CdTe, a CdTe photon-counting pixel array detector, has been used to measure the energy and the intensity of the white-beam diffraction from a lysozyme crystal. A method was developed to calibrate the detector in terms of energy, allowing incident photon energy measurement to high resolution (approximately 140 eV), opening up new possibilities in energy-resolved X-ray diffraction. In order to demonstrate this, Laue diffraction experiments were performed on the bending-magnet beamline METROLOGIE at Synchrotron SOLEIL. The X-ray energy spectra of diffracted spots were deduced from the indexed Laue patterns collected with an imaging-plate detector and then measured with both the XPAD3S-CdTe and the XPAD3S-Si, a silicon photon-counting pixel array detector. The predicted and measured energy of selected diffraction spots are in good agreement, demonstrating the reliability of the calibration method. These results open up the way to direct unit-cell parameter determination and the measurement of high-quality Laue data even at low resolution. Based on the success of these measurements, potential applications in X-ray diffraction opened up by this type of technology are discussed.

PCACE- PERSONAL COMPUTER AIDED CABLING ENGINEERING

NASA Technical Reports Server (NTRS)

Billitti, J. W.

1994-01-01

A computerized interactive harness engineering program has been developed to provide an inexpensive, interactive system which is designed for learning and using an engineering approach to interconnection systems. PCACE is basically a database system that stores information as files of individual connectors and handles wiring information in circuit groups stored as records. This directly emulates the typical manual engineering methods of data handling, thus making the user interface to the program very natural. Data files can be created, viewed, manipulated, or printed in real time. The printed ouput is in a form ready for use by fabrication and engineering personnel. PCACE also contains a wide variety of error-checking routines including connector contact checks during hardcopy generation. The user may edit existing harness data files or create new files. In creating a new file, the user is given the opportunity to insert all the connector and harness boiler plate data which would be part of a normal connector wiring diagram. This data includes the following: 1) connector reference designator, 2) connector part number, 3) backshell part number, 4) cable reference designator, 5) cable part number, 6) drawing revision, 7) relevant notes, 8) standard wire gauge, and 9) maximum circuit count. Any item except the maximum circuit count may be left blank, and any item may be changed at a later time. Once a file is created and organized, the user is directed to the main menu and has access to the file boiler plate, the circuit wiring records, and the wiring records index list. The organization of a file is such that record zero contains the connector/cable boiler plate, and all other records contain circuit wiring data. Each wiring record will handle a circuit with as many as nine wires in the interface. The record stores the circuit name and wire count and the following data for each wire: 1) wire identifier, 2) contact, 3) splice, 4) wire gauge if different from standard, 5) wire/group type, 6) wire destination, and 7) note number. The PCACE record structure allows for a wide variety of wiring forms using splices and shields, yet retains sufficient structure to maintain ease of use. PCACE is written in TURBO Pascal 3.0 and has been implemented on IBM PC, XT, and AT systems under DOS 3.1 with a memory of 512K of 8 bit bytes, two floppy disk drives, an RGB monitor, and a printer with ASCII control characters. PCACE was originally developed in 1983, and the IBM version was released in 1986.

Photon-counting image sensors for the ultraviolet

NASA Technical Reports Server (NTRS)

Jenkins, E. B.

1985-01-01

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

Microbial evaluation of Alaska salmon caviar.

PubMed

Himelbloom, B H; Crapo, C A

1998-05-01

Microbial quality of pink salmon caviar (ikura) processed at one plant in Alaska during a 30-day season was examined. Ikura (aw = 0.98; pH 6.1) averaged 49% water, 32% protein, 11% fat, 7% ash, and 3% salt. Aerobic plate counts (APCs) ranged from < 10(2)/g to 4.5 x 10(7)/g with increasing APC toward season's end. Coliform counts ranged from < 3/g to 2.4 x 10(3)/g. Escherichia coli, Staphylococcus aureus, yeasts, and molds were not detected. High-APC (10(7)/g) thawed caviar exhibited predominantly lactic acid bacteria; low-APC (10(3)/g) thawed caviar exhibited predominantly gram-negative bacteria. Freezing had little effect on the microbial counts, and shelf life of thawed caviar was 3 to 5 days at 2 degrees C.

Monitoring process hygiene in Serbian retail establishments

NASA Astrophysics Data System (ADS)

Vesković Moračanin, S.; Baltić, T.; Milojević, L.

2017-09-01

The present study was conducted to estimate the effectiveness of sanitary procedures on food contact surfaces and food handlers’ hands in Serbian retail establishments. For that purpose, a total of 970 samples from food contact surfaces and 525 samples from workers’ hands were microbiologically analyzed. Results of total aerobic plate count and total Enterobacteriaceae count showed that the implemented washing and disinfection procedures, as a part of HACCP plans, were not effective enough in most retail facilities. Constant and intensive education of employees on proper implementation of sanitation procedures are needed in order to ensure food safety in the retail market.

Photon-Counting H33D Detector for Biological Fluorescence Imaging

PubMed Central

Michalet, X.; Siegmund, O.H.W.; Vallerga, J.V.; Jelinsky, P.; Millaud, J.E.; Weiss, S.

2010-01-01

We have developed a photon-counting High-temporal and High-spatial resolution, High-throughput 3-Dimensional detector (H33D) for biological imaging of fluorescent samples. The design is based on a 25 mm diameter S20 photocathode followed by a 3-microchannel plate stack, and a cross delay line anode. We describe the bench performance of the H33D detector, as well as preliminary imaging results obtained with fluorescent beads, quantum dots and live cells and discuss applications of future generation detectors for single-molecule imaging and high-throughput study of biomolecular interactions. PMID:20151021

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

PubMed

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

2013-01-01

The purpose of this study was to assess the association of bulk tank milk standard plate counts, bulk tank coliform counts (CC), and the presence of Staphylococcus aureus in bulk tank milk with various management and farm characteristics on organic and conventional dairy farms throughout New York, Wisconsin, and Oregon. Data from size-matched organic farms (n=192), conventional nongrazing farms (n=64), and conventional grazing farms (n=36) were collected at a single visit for each farm. Of the 292 farms visited, 290 bulk tank milk samples were collected. Statistical models were created using data from all herds in the study, as well as exclusively for the organic subset of herds. Because of incomplete data, 267 of 290 herds were analyzed for total herd modeling, and 173 of 190 organic herds were analyzed for the organic herd modeling. Overall, more bulk tanks from organic farms had Staph. aureus cultured from them (62% of organic herds, 42% conventional nongrazing herds, and 43% of conventional grazing herds), whereas fewer organic herds had a high CC, defined as ≥50 cfu/mL, than conventional farms in the study. A high standard plate count (×1,000 cfu/mL) was associated with decreased body condition score of adult cows and decreased milk production in both models. Several variables were significant only in the model created using all herds or only in organic herds. The presence of Staph. aureus in the bulk tank milk was associated with fewer people treating mastitis, increased age of housing, and a higher percentage of cows with 3 or fewer teats in both the organic and total herd models. The Staph. aureus total herd model also showed a relationship with fewer first-lactation animals, higher hock scores, and less use of automatic takeoffs at milking. High bulk tank CC was related to feeding a total mixed ration and using natural service in nonlactating heifers in both models. Overall, attentive management and use of outside resources were useful with regard to CC on organic farms. In all models except the organic CC model, we observed an association with the average reported somatic cell count from 3 mo before the herd visit, indicating that many of the regularly tested milk quality parameters are interconnected. In conclusion, we found that conventional and organic farms are similar in regard to overall herd management, but each grazing system faces unique challenges when managing milk quality. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

PubMed

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

2008-10-31

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

Survival of B. Horneckiae Spores Under Ground-simulated Space Conditions

NASA Technical Reports Server (NTRS)

Schanche, Bradley

2012-01-01

To prevent forward contamination and maintain the scientific integrity of future life detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated habitats, spore-forming microbes are highly resistant to various physical and chemical conditions, which include ionizing and UV radiation, desiccation and oxidative stress, and the harsh environment of outer space or planetary surfaces. Recently a radiation resistant, spore forming bacterial isolate, Bacillus horneckiae, was isolated from a clean room of the Kennedy Space Center where the Phoenix spacecraft was assembled. The exceptionally high tolerance of extreme conditions demonstrated by sporeforming bacteria highlighted the need to assess the viability of these microbes in situ (in real) space. The proposed BOSS (Biofilm Organisms Surfing Space) project aims to understand the mechanisms by which biofilm forming organisms, such as B. horneckiae, will potentially be able to withstand harsh space conditions. As previously stated, the spore producing ability of these species gives them increased survivability to harsh conditions. Some of the spores will have the protective exosporium layer artificially removed before the test to determine if the existence of this layer significantly changes the survivability during the mission. In preparation for that experiment, we analyzed spores which were exposed during a ground simulation, the EXPOSE R2 Biofilm Organisms Surfing Space (BOSS). Previous to exposure, spores were deposited onto spacecraft grade aluminum coupons in a spore suspension calculated to contain between 10(exp 7) and 10(exp 8) spores. This precursor series will be used to establish a baseline survivability function for comparison with the future flight tests during EXPOSE-R. For each coupon, a 10% polyvinyl alcohol (PVA) film was applied and peeled from the coupon to recover the spores. One hundred µl of sterile 10% PVA was applied to the surface of the coupon and allowed to dry for 1 hour at 37 C. The films were then removed using sterile scalpel and forceps and placed into a glass test tube containing 2 milliliters of sterile deionized water. The PVA film process was then repeated on each coupon one additional time to ensure recovery of the majority of spores. The second PVA film was added in the same glass tube as in the previous round. If the spores remained 100% viable, the test tubes should now contain between 5 X 10(exp 6) and 5 X 10(exp 7) spores per millimeter; however, it is expected that some loss of viability has occurred. In order to assess this loss, the number of colony forming, viable spores was counted. To count the colony forming units (CFUs), the spore containing solution was diluted in a process of 10-fold serial dilution by mixing successive solutions in a 100 microliter spore suspension to 900 microliter deionized H2O ratio. A sample dilution series revealed that 10(exp -3) and 10(exp -4) concentrations would be necessary for an accurate CFU count to be taken. For those two concentrations, a spread on a TSA plate was prepared and incubated at 32 C. For the samples exposed to UV radiation, the cell survivability was too low to establish a count from 100 microliter spread plating. Instead, no dilutions were performed and the entire 2 milliliter spore suspension was plated and incubated at 32 C. The plate's CFU counts were taken at 24 hours and 48 hours from the time of plating. At the end of the CFU counting the total surviving spores in each sample were calculated based on the number of CFUs that were observed per 100 microliters, or per 2 milliliters for the UV irradiated samples. The results of these calculations are shown in Figures 1 and 2.

New semi-pilot-scale reactor to study the photocatalytic inactivation of phages contained in aerosol.

PubMed

Briggiler Marcó, Mariángeles; Negro, Antonio Carlos; Alfano, Orlando Mario; Quiberoni, Andrea Del Luján

2017-04-12

The aims of this work were to design and build a photocatalytic reactor (UV-A/TiO 2 ) to study the inactivation of phages contained in bioaerosols, which constitute the main dissemination via phages in industrial environments. The reactor is a close system with recirculation that consists of a stainless steel camera (cubic form, side of 60 cm) in which air containing the phage particles circulates and an acrylic compartment with six borosilicate plates covered with TiO 2 . The reactor is externally illuminated by 20 UV-A lamps. Both compartments are connected by a fan to facilitate the sample circulation. Samples are injected into the camera using two piston nebulizers working in series whereas several methodologies for sampling (impinger/syringe, sampling on photocatalytic plates, and impact of air on slide) were assayed. The reactor setup was carried out using phage B1 (Lactobacillus plantarum), and assays demonstrated a decrease of phage counts of 2.7 log orders after 1 h of photocatalytic treatment. Photonic efficiencies of inactivation were assessed by phage sampling on the photocatalytic plates or by impact of air on a glass slide at the photocatalytic reactor exit. Efficiencies of the same order of magnitude were observed using both sampling methods. This study demonstrated that the designed photocatalytic reactor is effective to inactivate phage B1 (Lb. plantarum) contained in bioaerosols.

Effects of a hybrid micro/nanorod topography-modified titanium implant on adhesion and osteogenic differentiation in rat bone marrow mesenchymal stem cells.

PubMed

Zhang, Wenjie; Li, Zihui; Huang, Qingfeng; Xu, Ling; Li, Jinhua; Jin, Yuqin; Wang, Guifang; Liu, Xuanyong; Jiang, Xinquan

2013-01-01

Various methods have been used to modify titanium implant surfaces with the aim of achieving better osseointegration. In this study, we fabricated a clustered nanorod structure on an acid-etched, microstructured titanium plate surface using hydrogen peroxide. We also evaluated biofunctionalization of the hybrid micro/nanorod topography on rat bone marrow mesenchymal stem cells. Scanning electron microscopy and x-ray diffraction were used to investigate the surface topography and phase composition of the modified titanium plate. Rat bone marrow mesenchymal stem cells were cultured and seeded on the plate. The adhesion ability of the cells was then assayed by cell counting at one, 4, and 24 hours after cell seeding, and expression of adhesion-related protein integrin β1 was detected by immunofluorescence. In addition, a polymerase chain reaction assay, alkaline phosphatase and Alizarin Red S staining assays, and osteopontin and osteocalcin immunofluorescence analyses were used to evaluate the osteogenic differentiation behavior of the cells. The hybrid micro/nanoscale texture formed on the titanium surface enhanced the initial adhesion activity of the rat bone marrow mesenchymal stem cells. Importantly, the hierarchical structure promoted osteogenic differentiation of these cells. This study suggests that a hybrid micro/nanorod topography on a titanium surface fabricated by treatment with hydrogen peroxide followed by acid etching might facilitate osseointegration of a titanium implant in vivo.

Osteosynthesis using plates and screws after removing a limited area of the periosteum in order to reduce misclassified during radiological assessment metacarpal shaft fractures

PubMed Central

Neagu, TP; Popescu, SA; Cobilinschi, C; Tincu, R; Tiglis, M; Lascar, I

2016-01-01

Hand fractures are one of the most common causes for presenting to the emergency room. Metacarpal fractures count about 18 to 44% of all hand fractures, and are most often standalone closed injuries, without misplacement, not needing operative treatment. We present a case in which osteosynthesis with plates and screws was used to reduce two metacarpal fractures in order to allow an early motion recovery, despite the fact that a small portion of the periosteum needed to be removed. The type of fractures were misclassified according to the radiological findings, therefore the correct diagnosis was established during surgery. The results according to the radiological aspects and to the DASH score were excellent with 95% function recovery at twelve months. In this case, the use of osteosynthesis with plates and screws led to a good fracture healing without any major complications. However, there are a series of complications related to this method that should be taken into consideration. Being misled by the radiological aspects of the fractures, the most certain way to classify a metacarpal shaft fracture is through exploratory surgery, even if in most of the cases the three radiological views are enough to establish the diagnosis. Abbreviations: DASH score = Disability of Arm, Shoulder and Hand score, TAM = Total Active Motion, MCP = metacarpal phalangeal joint, PIP = proximal inter phalangeal joint PMID:27974942

Inter-operator variation in ELISPOT analysis of measles virus-specific IFN-gamma-secreting T cells.

PubMed

Ryan, J E; Ovsyannikova, I G; Dhiman, N; Pinsky, N A; Vierkant, R A; Jacobson, R M; Poland, G A

2005-01-01

The ELISPOT assay is a highly sensitive technique used for the detection of individual cytokine releasing cells. We have developed an IFN-gamma ELISPOT assay utilizing unfractionated frozen peripheral blood mononuclear cells (PBMC) to quantify the frequency of measles virus (MV)-specific IFN-gamma-secreting T cells in 117 healthy children who had been previously immunized with two doses of the measles-mumps-rubella vaccine. We have also estimated the variability associated with the quantification of ELISPOT plates and compared the number of MV-specific IFN-gamma-secreting T cells for each subject as determined by two different operators of an ELISPOT reader. The median frequency of MV-specific IFN-gamma-producing memory T cells detected by this assay was 0.005 % and 0.01 % as determined by an in-house and commercial operator, respectively. Although we found a significant correlation (r = 0.83, p<0.0001) between the number of spots counted by the commercial and in-house operators of an ELISPOT reader, the median number of spots counted by the commercial operator was twice the number of spots counted by an in-house operator (p<0.001). This demonstrates the importance of using a common ELISPOT reader and operator, among other parameters, to quantify the number of spots when a large volume of plates are being scanned and analyzed.

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

PubMed

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.

Stress-opioid interactions: a comparison of morphine and methadone.

PubMed

Taracha, Ewa; Mierzejewski, Paweł; Lehner, Małgorzata; Chrapusta, Stanisław J; Kała, Maria; Lechowicz, Wojciech; Hamed, Adam; Skórzewska, Anna; Kostowski, Wojciech; Płaźnik, Adam

2009-01-01

The utility of methadone and morphine for analgesia and of methadone for substitution therapy for heroin addiction is a consequence of these drugs acting as opioid receptor agonists.We compared the cataleptogenic and antinociceptive effects of single subcutaneous doses of methadone hydrochloride (1-4 mg/kg) and morphine sulfate (2.5-10 mg/kg) using catalepsy and hot-plate tests, and examined the effects of the highest doses of the drugs on Fos protein expression in selected brain regions in male Sprague-Dawley rats. Methadone had greater cataleptogenic and analgesic potency than morphine. Fos immunohistochemistry revealed substantial effects on the Fos response of both the stress induced by the experimental procedures and of the drug exposure itself. There were three response patterns identified: 1) drug exposure, but not stress, significantly elevated Fos-positive cell counts in the caudate-putamen; 2) stress alone and stress combined with drug exposure similarly elevated Fos-positive cell counts in the nucleus accumbens and cingulate cortex; and 3) methadone and morphine (to a lesser extent) counteracted the stimulatory effect of nonpharmacological stressors on Fos protein expression in the somatosensory cortex barrel field, and Fos-positive cell counts in this region correlated negatively with both the duration of catalepsy and the latency time in the hot-plate test. The overlap between brain regions reacting to nonpharmacological stressors and those responding to exogenous opioids suggests that stress contributes to opioid-induced neuronal activation.

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

PubMed

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.

High throughput and quantitative approaches for measuring circadian rhythms in cyanobacteria using bioluminescence

PubMed Central

Shultzaberger, Ryan K.; Paddock, Mark L.; Katsuki, Takeo; Greenspan, Ralph J.; Golden, Susan S.

2016-01-01

The temporal measurement of a bioluminescent reporter has proven to be one of the most powerful tools for characterizing circadian rhythms in the cyanobacterium Synechococcus elongatus. Primarily, two approaches have been used to automate this process: (1) detection of cell culture bioluminescence in 96-well plates by a photomultiplier tube-based plate-cycling luminometer (TopCount Microplate Scintillation and Luminescence Counter, Perkin Elmer) and (2) detection of individual colony bioluminescence by iteratively rotating a Petri dish under a cooled CCD camera using a computer-controlled turntable. Each approach has distinct advantages. The TopCount provides a more quantitative measurement of bioluminescence, enabling the direct comparison of clock output levels among strains. The computer-controlled turntable approach has a shorter set-up time and greater throughput, making it a more powerful phenotypic screening tool. While the latter approach is extremely useful, only a few labs have been able to build such an apparatus because of technical hurdles involved in coordinating and controlling both the camera and the turntable, and in processing the resulting images. This protocol provides instructions on how to construct, use, and process data from a computer-controlled turntable to measure the temporal changes in bioluminescence of individual cyanobacterial colonies. Furthermore, we describe how to prepare samples for use with the TopCount to minimize experimental noise, and generate meaningful quantitative measurements of clock output levels for advanced analysis. PMID:25662451

SkICAT: A cataloging and analysis tool for wide field imaging surveys

NASA Technical Reports Server (NTRS)

Weir, N.; Fayyad, U. M.; Djorgovski, S. G.; Roden, J.

1992-01-01

We describe an integrated system, SkICAT (Sky Image Cataloging and Analysis Tool), for the automated reduction and analysis of the Palomar Observatory-ST ScI Digitized Sky Survey. The Survey will consist of the complete digitization of the photographic Second Palomar Observatory Sky Survey (POSS-II) in three bands, comprising nearly three Terabytes of pixel data. SkICAT applies a combination of existing packages, including FOCAS for basic image detection and measurement and SAS for database management, as well as custom software, to the task of managing this wealth of data. One of the most novel aspects of the system is its method of object classification. Using state-of-theart machine learning classification techniques (GID3* and O-BTree), we have developed a powerful method for automatically distinguishing point sources from non-point sources and artifacts, achieving comparably accurate discrimination a full magnitude fainter than in previous Schmidt plate surveys. The learning algorithms produce decision trees for classification by examining instances of objects classified by eye on both plate and higher quality CCD data. The same techniques will be applied to perform higher-level object classification (e.g., of galaxy morphology) in the near future. Another key feature of the system is the facility to integrate the catalogs from multiple plates (and portions thereof) to construct a single catalog of uniform calibration and quality down to the faintest limits of the survey. SkICAT also provides a variety of data analysis and exploration tools for the scientific utilization of the resulting catalogs. We include initial results of applying this system to measure the counts and distribution of galaxies in two bands down to Bj is approximately 21 mag over an approximate 70 square degree multi-plate field from POSS-II. SkICAT is constructed in a modular and general fashion and should be readily adaptable to other large-scale imaging surveys.

Effect of salt concentrations and drying methods on the quality and formation of histamine in dried milkfish (Chanos chanos).

PubMed

Hwang, Chiu-Chu; Lin, Chia-Min; Kung, Hsien-Feng; Huang, Ya-Ling; Hwang, Deng-Fwu; Su, Yi-Cheng; Tsai, Yung-Hsiang

2012-11-15

The effects of salt concentrations (0-15.0%) and drying methods on the quality of dried milkfish were studied. The results showed that the levels of aerobic plate counts, total coliform, water activity, moisture contents, total volatile basic nitrogen (TVBN) and thiobarbituric acid (TBA) of the dried milkfish samples prepared with the same drying method decreased with increased salt concentrations. The samples prepared with the cold-air drying method had better quality in term of lower TVBN and TBA values than those of samples prepared with other drying methods. The histamine contents in all samples, except two, prepared with various salt concentrations by different drying methods were less than 1.9 mg/100 g. Two unsalted samples prepared with hot-air drying at 35 °C and sun drying methods were found to contain histamine at levels of 249.7 and 67.4 mg/100 g, respectively, which were higher than the potential hazard level of 50 mg/100 g. Copyright © 2012 Elsevier Ltd. All rights reserved.

Survival of cold-stressed Campylobacter jejuni on ground chicken and chicken skin during frozen storage.

PubMed

Bhaduri, Saumya; Cottrell, Bryan

2004-12-01

Campylobacter jejuni is prevalent in poultry, but the effect of combined refrigerated and frozen storage on its survival, conditions relevant to poultry processing and storage, has not been evaluated. Therefore, the effects of refrigeration at 4 degrees C, freezing at -20 degrees C, and a combination of refrigeration and freezing on the survival of C. jejuni in ground chicken and on chicken skin were examined. Samples were enumerated using tryptic soy agar containing sheep's blood and modified cefoperazone charcoal deoxycholate agar. Refrigerated storage alone for 3 to 7 days produced a reduction in cell counts of 0.34 to 0.81 log10 CFU/g in ground chicken and a reduction in cell counts of 0.31 to 0.63 log10 CFU/g on chicken skin. Declines were comparable for each sample type using either plating medium. Frozen storage, alone and with prerefrigeration, produced a reduction in cell counts of 0.56 to 1.57 log10 CFU/g in ground chicken and a reduction in cell counts of 1.38 to 3.39 log10 CFU/g on chicken skin over a 2-week period. The recovery of C. jejuni following freezing was similar on both plating media. The survival following frozen storage was greater in ground chicken than on chicken skin with or without prerefrigeration. Cell counts after freezing were lower on chicken skin samples that had been prerefrigerated for 7 days than in those that had been prerefrigerated for 0, 1, or 3 days. This was not observed for ground chicken samples, possibly due to their composition. C. jejuni survived storage at 4 and -20 degrees C with either sample type. This study indicates that, individually or in combination, refrigeration and freezing are not a substitute for safe handling and proper cooking of poultry.

A Spore Counting Method and Cell Culture Model for Chlorine Disinfection Studies of Encephalitozoon syn. Septata intestinalis

PubMed Central

Wolk, D. M.; Johnson, C. H.; Rice, E. W.; Marshall, M. M.; Grahn, K. F.; Plummer, C. B.; Sterling, C. R.

2000-01-01

The microsporidia have recently been recognized as a group of pathogens that have potential for waterborne transmission; however, little is known about the effects of routine disinfection on microsporidian spore viability. In this study, in vitro growth of Encephalitozoon syn. Septata intestinalis, a microsporidium found in the human gut, was used as a model to assess the effect of chlorine on the infectivity and viability of microsporidian spores. Spore inoculum concentrations were determined by using spectrophotometric measurements (percent transmittance at 625 nm) and by traditional hemacytometer counting. To determine quantitative dose-response data for spore infectivity, we optimized a rabbit kidney cell culture system in 24-well plates, which facilitated calculation of a 50% tissue culture infective dose (TCID50) and a minimal infective dose (MID) for E. intestinalis. The TCID50 is a quantitative measure of infectivity and growth and is the number of organisms that must be present to infect 50% of the cell culture wells tested. The MID is as a measure of a system's permissiveness to infection and a measure of spore infectivity. A standardized MID and a standardized TCID50 have not been reported previously for any microsporidian species. Both types of doses are reported in this paper, and the values were used to evaluate the effects of chlorine disinfection on the in vitro growth of microsporidia. Spores were treated with chlorine at concentrations of 0, 1, 2, 5, and 10 mg/liter. The exposure times ranged from 0 to 80 min at 25°C and pH 7. MID data for E. intestinalis were compared before and after chlorine disinfection. A 3-log reduction (99.9% inhibition) in the E. intestinalis MID was observed at a chlorine concentration of 2 mg/liter after a minimum exposure time of 16 min. The log10 reduction results based on percent transmittance-derived spore counts were equivalent to the results based on hemacytometer-derived spore counts. Our data suggest that chlorine treatment may be an effective water treatment for E. intestinalis and that spectrophotometric methods may be substituted for labor-intensive hemacytometer methods when spores are counted in laboratory-based chlorine disinfection studies. PMID:10742198

The ultraviolet detection component based on Te-Cs image intensifier

NASA Astrophysics Data System (ADS)

Qian, Yunsheng; Zhou, Xiaoyu; Wu, Yujing; Wang, Yan; Xu, Hua

2017-05-01

Ultraviolet detection technology has been widely focused and adopted in the fields of ultraviolet warning and corona detection for its significant value and practical meaning. The component structure of ultraviolet ICMOS, imaging driving and the photon counting algorithm are studied in this paper. Firstly, the one-inch and wide dynamic range CMOS chip with the coupling optical fiber panel is coupled to the ultraviolet image intensifier. The photocathode material in ultraviolet image intensifier is Te-Cs, which contributes to the solar blind characteristic, and the dual micro-channel plates (MCP) structure ensures the sufficient gain to achieve the single photon counting. Then, in consideration of the ultraviolet detection demand, the drive circuit of the CMOS chip is designed and the corresponding program based on Verilog language is written. According to the characteristics of ultraviolet imaging, the histogram equalization method is applied to enhance the ultraviolet image and the connected components labeling way is utilized for the ultraviolet single photon counting. Moreover, one visible light video channel is reserved in the ultraviolet ICOMS camera, which can be used for the fusion of ultraviolet and visible images. Based upon the module, the ultraviolet optical lens and the deep cut-off solar blind filter are adopted to construct the ultraviolet detector. At last, the detection experiment of the single photon signal is carried out, and the test results are given and analyzed.

Microbiota during fermentation of chum salmon (Oncorhynchus keta) sauce mash inoculated with halotolerant microbial starters: analyses using the plate count method and PCR-denaturing gradient gel electrophoresis (DGGE).

PubMed

Yoshikawa, Shuji; Yasokawa, Daisuke; Nagashima, Koji; Yamazaki, Koji; Kurihara, Hideyuki; Ohta, Tomoki; Kawai, Yuji

2010-06-01

Nine different combinations of mugi koji (barley steamed and molded with Aspergillus oryzae) and halotolerant microorganisms (HTMs), Zygosaccharomyces rouxii, Candida versatilis, and Tetragenococcus halophilus, were inoculated into chum salmon sauce mash under a non-aseptic condition used in industrial fish sauce production and fermented at 35 +/- 2.5 degrees C for 84 days to elucidate the microbial dynamics (i.e., microbial count and microbiota) during fermentation. The viable count of halotolerant yeast (HTY) in fermented chum salmon sauce (FCSS) mash showed various time courses dependent on the combination of the starter microorganisms. Halotolerant lactic acid bacteria (HTL) were detected morphologically and physiologically only from FCSS mash inoculated with T. halophilus alone or with T. halophilus and C. versatilis during the first 28 days of fermentation. Only four fungal species, Z. rouxii, C. versatilis, Pichia guilliermondii, and A. oryzae, were detected throughout the fermentation by PCR-denaturing gradient gel electrophoresis (PCR-DGGE). In FCSS mash, dominant HTMs, especially eumycetes, were nonexistent. However, under the non-aseptic conditions, undesirable wild yeast such as P. guilliermondii grew fortuitously. Therefore, HTY inoculation into FCSS mash at the beginning of fermentation is effective in preventing the growth of wild yeast and the resultant unfavorable flavor. 2009 Elsevier Ltd. All rights reserved.

Use of non-linear mixed-effects modelling and regression analysis to predict the number of somatic coliphages by plaque enumeration after 3 hours of incubation.

PubMed

Mendez, Javier; Monleon-Getino, Antonio; Jofre, Juan; Lucena, Francisco

2017-10-01

The present study aimed to establish the kinetics of the appearance of coliphage plaques using the double agar layer titration technique to evaluate the feasibility of using traditional coliphage plaque forming unit (PFU) enumeration as a rapid quantification method. Repeated measurements of the appearance of plaques of coliphages titrated according to ISO 10705-2 at different times were analysed using non-linear mixed-effects regression to determine the most suitable model of their appearance kinetics. Although this model is adequate, to simplify its applicability two linear models were developed to predict the numbers of coliphages reliably, using the PFU counts as determined by the ISO after only 3 hours of incubation. One linear model, when the number of plaques detected was between 4 and 26 PFU after 3 hours, had a linear fit of: (1.48 × Counts 3 h + 1.97); and the other, values >26 PFU, had a fit of (1.18 × Counts 3 h + 2.95). If the number of plaques detected was <4 PFU after 3 hours, we recommend incubation for (18 ± 3) hours. The study indicates that the traditional coliphage plating technique has a reasonable potential to provide results in a single working day without the need to invest in additional laboratory equipment.

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

PubMed

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

2018-03-01

The objective of this study was to measure the effect of different milk cooling rates, before entering the bulk tank, on the microbiological load and composition of the milk, as well as on energy usage. Three milk precooling treatments were applied before milk entered 3 identical bulk milk tanks: no plate cooler (NP), single-stage plate cooler (SP), and double-stage plate cooler (DP). These precooling treatments cooled the milk to 32.0 ± 1.4°C, 17.0 ± 2.8°C, and 6.0 ± 1.1°C, respectively. Milk was added to the bulk tank twice daily for 72 h, and the tank refrigeration temperature was set at 3°C. The blend temperature within each bulk tank was reduced after each milking event as the volume of milk at 3°C increased simultaneously. The bacterial counts of the milk volumes precooled at different rates did not differ significantly at 0 h of storage or at 24-h intervals thereafter. After 72 h of storage, the total bacterial count of the NP milk was 3.90 ± 0.09 log 10 cfu/mL, whereas that of the precooled milk volumes were 3.77 ± 0.09 (SP) and 3.71 ± 0.09 (DP) log 10 cfu/mL. The constant storage temperature (3°C) over 72 h helped to reduce bacterial growth rates in milk; consequently, milk composition was not affected and minimal, if any, proteolysis occurred. The DP treatment had the highest energy consumption (17.6 ± 0.5 Wh/L), followed by the NP (16.8 ± 2.7 Wh/L) and SP (10.6 ± 1.3 Wh/L) treatments. This study suggests that bacterial count and composition of milk are minimally affected when milk is stored at 3°C for 72 h, regardless of whether the milk is precooled; however, milk entering the tank should have good initial microbiological quality. Considering the numerical differences between bacterial counts, however, the use of the SP or DP precooling systems is recommended to maintain low levels of bacterial counts and reduce energy consumption. The Authors. Published by FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

Multi-anode microchannel arrays

NASA Technical Reports Server (NTRS)

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

1977-01-01

A development program is currently being undertaken to produce photon-counting detector arrays which are suitable for use in both ground-based and space-borne instruments and which utilize the full sensitivity, dynamic range and photometric stability of the microchannel array plate (MCP). The construction of the detector arrays and the status of the development program are described.

Paper Plate Fractions: The Counting Connection

ERIC Educational Resources Information Center

McCoy, Ann; Barnett, Joann; Stine, Tammy

2016-01-01

Without a doubt, fractions prove to be a stumbling block for many children. Researchers have suggested a variety of explanations for why this is the case. The introduction of symbolization and operations before the development of conceptual understanding of fractions, a lack of understanding of the role of the numerator and denominator, and an…

Bacteria associated with granular activated carbon particles in drinking water.

PubMed Central

Camper, A K; LeChevallier, M W; Broadaway, S C; McFeters, G A

1986-01-01

A sampling protocol was developed to examine particles released from granular activated carbon filter beds. A gauze filter/Swinnex procedure was used to collect carbon fines from 201 granular activated carbon-treated drinking water samples over 12 months. Application of a homogenization procedure (developed previously) indicated that 41.4% of the water samples had heterotrophic plate count bacteria attached to carbon particles. With the enumeration procedures described, heterotrophic plate count bacteria were recovered at an average rate of 8.6 times higher than by conventional analyses. Over 17% of the samples contained carbon particles colonized with coliform bacteria as enumerated with modified most-probable-number and membrane filter techniques. In some instances coliform recoveries were 122 to 1,194 times higher than by standard procedures. Nearly 28% of the coliforms attached to these particles in drinking water exhibited the fecal biotype. Scanning electron micrographs of carbon fines from treated drinking water showed microcolonies of bacteria on particle surfaces. These data indicate that bacteria attached to carbon fines may be an important mechanism by which microorganisms penetrate treatment barriers and enter potable water supplies. PMID:3767356

Growth and persistence of pathogens on granular activated carbon filters.

PubMed Central

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

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

PubMed

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

2011-12-01

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

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

PubMed

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

2015-01-01

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

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

NASA Technical Reports Server (NTRS)

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

2001-01-01

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

Relative Numbers of Certain Microbial Groups Present in Compost Used for Mushroom (Agaricus bisporus) Propagation

PubMed Central

Fordyce, C.

1970-01-01

The relative numbers of microorganisms associated with compost during mushroom production were studied by the dilution plate method. Thermophilic actinomycetes and fungi were isolated with a very high frequency early in the growing season. Although numbers of thermophilic bacteria diminished slowly during the season, the thermophilic fungi and actinomycetes diminished rapidly with the latter disappearing after 6 weeks. Mesophilic fungi other than Agaricus or Trichoderma remained relatively stable throughout the growing period. Agaricus could be isolated between the first and third break. Trichoderma became dominant after the fourth break. The mesophilic bacterial counts diminished during the most productive portion of the mushroom cropping season and then increased to much higher numbers toward the end of the season. PMID:5529631

Removing Escherichia coli from water using zinc oxide-coated zeolite.

PubMed

Wang, Lingling; Wu, Wenlin; Xie, Xiaolan; Chen, Hongbin; Lin, Jianming; Dionysiou, Dionysios D

2018-05-11

The removal of Escherichia coli (E. coli) from water by zinc oxide-coated zeolite (ZOCZ) and ZOCZ's antibacterial properties were examined in laboratory experiments using plate counting method and tests of cell apoptosis. Batch experiments showed that ZOCZ has a maximum removal capacity for E. coli of about 4.34 × 10 6  CFU g -1  at 25 °C. Element mappings confirm that zinc ions accumulate in the E. coli cells causing cell death. Pseudo-second-order kinetics and Freundlich isotherms were found to best describe the removal of E. coli, suggesting that a multilayer of E. coli cells forms on the surface of ZOCZ particles. Copyright © 2018 Elsevier Ltd. All rights reserved.

The effect of ice-cream-scoop water on the hygiene of ice cream.

PubMed Central

Wilson, I. G.; Heaney, J. C.; Weatherup, S. T.

1997-01-01

A survey of unopened ice cream, ice cream in use, and ice-cream-scoop water (n = 91) was conducted to determine the effect of scoop water hygiene on the microbiological quality of ice cream. An aerobic plate count around 10(6) c.f.u. ml-1 was the modal value for scoop waters. Unopened ice creams generally had counts around 10(3)-10(4) c.f.u. ml-1 and this increased by one order of magnitude when in use. Many scoop waters had low coliform counts, but almost half contained > 100 c.f.u. ml-1. E. coli was isolated in 18% of ice creams in use, and in 10% of unopened ice creams. S. aureus was not detected in any sample. Statistical analysis showed strong associations between indicator organisms and increased counts in ice cream in use. EC guidelines for indicator organisms in ice cream were exceeded by up to 56% of samples. PMID:9287941

The effect of ice-cream-scoop water on the hygiene of ice cream.

PubMed

Wilson, I G; Heaney, J C; Weatherup, S T

1997-08-01

A survey of unopened ice cream, ice cream in use, and ice-cream-scoop water (n = 91) was conducted to determine the effect of scoop water hygiene on the microbiological quality of ice cream. An aerobic plate count around 10(6) c.f.u. ml-1 was the modal value for scoop waters. Unopened ice creams generally had counts around 10(3)-10(4) c.f.u. ml-1 and this increased by one order of magnitude when in use. Many scoop waters had low coliform counts, but almost half contained > 100 c.f.u. ml-1. E. coli was isolated in 18% of ice creams in use, and in 10% of unopened ice creams. S. aureus was not detected in any sample. Statistical analysis showed strong associations between indicator organisms and increased counts in ice cream in use. EC guidelines for indicator organisms in ice cream were exceeded by up to 56% of samples.

The Application of Active Paper Incorporated with Oleoresin of Cinnamon Leaf (Cinnamomum burmanii) Distillation Residues on Maintaining Dragon Fruits (Hylocereus costaricensis) Quality during Storage

NASA Astrophysics Data System (ADS)

Aziz, M. S. H.; Manuhara, G. J.; Utami, R.; Khasanah, L. U.

2018-03-01

The purpose of this study was to determine the effect of active paper placement methods on super red dragon fruits quality during storage at ambient temperature. The active papers were incorporated with oleoresin of cinnamon leaf distillation residues. Various active paper placement methods were applied such as wrapping, placed on the cardboard wall, placed cardboard pad, and scrap of paper on the sidelines. Weight loss, peel color, surface and flesh hardness, total titratable acid, soluble solid total, pH flesh fruit, and total plate count (TPC) of super red dragon fruits samples were investigated during 9 days storage. The result shows that active paper placement methods significantly affected the weight loss, surface firmness and color peel change of super red dragon fruits samples. However, active paper placement methods insignificantly affected the titrable acid total, soluble solid total, pH, flesh firmness and microbial spoilage of super red dragon fruits samples. The best method to maintain the super red dragon fruits quality was wrapping method.

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

PubMed

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

2016-08-01

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

MRP8/14 Enhances Corneal Susceptibility to Pseudomonas aeruginosa Infection by Amplifying Inflammatory Responses

PubMed Central

Deng, Qiuchan; Sun, Mingxia; Yang, Kun; Zhu, Min; Chen, Kang; Yuan, Jin; Wu, Minhao; Huang, Xi

2013-01-01

Purpose. We explored the role of myeloid-related protein 8 and 14 (MRP8/14) in Pseudomonas aeruginosa (PA) keratitis. Methods. MRP8/14 mRNA levels in human corneal scrapes and mouse corneas infected by PA were tested using real-time PCR. MRP8/14 protein expression in C57BL/6 (B6) corneas was confirmed using Western blot assay and immunohistochemistry. B6 mice were injected subconjunctivally with siRNA for MRP8/14, and then infected with PA. Bacterial plate counts and myeloperoxidase assays were used to determine the bacterial load and polymorphonuclear neutrophil (PMN) infiltration in infected B6 corneas. Pro-inflammatory cytokine levels in vivo and in vitro were examined with PCR and ELISA. In murine macrophage-like RAW264.7 cells, phagocytosis and bacterial killing were assessed using plate count assays, and reactive oxygen species (ROS) and nitric oxide (NO) levels were tested with flow cytometry and Griess assay, respectively. Results. MRP8/14 expression levels were increased significantly in human corneal scrapes and B6 corneas after PA infection. Silencing of MRP8/14 in B6 corneas significantly reduced the severity of corneal disease, bacterial clearance, PMN infiltration, and pro-inflammatory cytokine expression after PA infection. In vitro studies demonstrated further that silencing of MRP8/14 suppressed pro-inflammatory cytokine production, bacterial killing, and ROS production, but not phagocytosis or NO production. Conclusions. Our study demonstrated a dual role for MRP8/14 in bacterial keratitis. Although MRP8/14 promotes bacterial clearance by enhancing ROS production, it functions more importantly as an inflammatory amplifier at the ocular surface by enhancing pro-inflammatory cytokine expression, thus contributing to the corneal susceptibility. PMID:23299480

Comparison of a Four-Section Spindle and Stomacher for Efficacy of Detaching Microorganisms from Fresh Vegetables.

PubMed

Kim, Do-Kyun; Kim, Soo-Ji; Kang, Dong-Hyun

2015-07-01

This study was undertaken to compare the effect of the spindle and stomacher for detaching microorganisms from fresh vegetables. The spindle is an apparatus for detaching microorganisms from food surfaces, which was developed in our laboratory. When processed with the spindle, food samples were barely disrupted, the original shape was maintained, and the diluent was clear, facilitating further detection analysis more easily than with stomacher treatment. The four-section spindle consists of four sample bag containers (A, B, C, and D) to economize time and effort by simultaneously processing four samples. The aerobic plate counts (APC) of 50 fresh vegetable samples were measured following spindle and stomacher treatment. Correlations between the two methods for each section of the spindle and stomacher were very high (R(2) = 0.9828 [spindle compartment A; Sp A], 0.9855 [Sp B], 0.9848 [Sp C], and 0.9851 [Sp D]). One-tenth milliliter of foodborne pathogens suspensions was inoculated onto surfaces of food samples, and ratios of spindle-to-stomacher enumerations were close to 1.00 log CFU/g between every section of the spindle and stomacher. One of the greatest features of the spindle is that it can treat large-sized samples that exceed 200 g. Uncut whole apples, green peppers, potatoes, and tomatoes were processed by the spindle and by hand massaging by 2 min. Large-sized samples were also assayed for aerobic plate count and recovery of the three foodborne pathogens, and the difference between each section of the spindle and hand massaging was not significant (P > 0.05). This study demonstrated that the spindle apparatus can be an alternative device for detaching microorganisms from all fresh vegetable samples for microbiological analysis by the food processing industry.

Development of multiplex PCR for the detection of total coliform bacteria for Escherichia coli and Clostridium perfringens in drinking water.

PubMed

Tantawiwat, Suwalee; Tansuphasiri, Unchalee; Wongwit, Waranya; Wongchotigul, Varee; Kitayaporn, Dwip

2005-01-01

Multiplex PCR amplification of lacZ, uidA and plc genes was developed for the simultaneous detection of total coliform bacteria for Escherichia coli and Clostridium perfringens, in drinking water. Detection by agarose gel electrophoresis yielded a band of 876 bp for the lacZ gene of all coliform bacteria; a band of 147 bp for the uidA gene and a band of 876 bp for the lacZ gene of all strains of E. coli; a band of 280 bp for the p/c gene for all strains of C. perfringens; and a negative result for all three genes when tested with other bacteria. The detection limit was 100 pg for E. coli and C. perfringens, and 1 ng for coliform bacteria when measured with purified DNA. This assay was applied to the detection of these bacteria in spiked water samples. Spiked water samples with 0-1,000 CFU/ml of coliform bacteria and/or E. coli and/or C. perfringens were detected by this multiplex PCR after a pre-enrichment step to increase the sensitivity and to ensure that the detection was based on the presence of cultivable bacteria. The result of bacterial detection from the multiplex PCR was comparable with that of a standard plate count on selective medium (p=0.62). When using standard plate counts as a gold standard, the sensitivity for this test was 99.1% (95% CI 95.33, 99.98) and the specificity was 90.9 % (95% CI 75.67, 98.08). Multiplex PCR amplification with a pre-enrichment step was shown to be an effective, sensitive and rapid method for the simultaneous detection of these three microbiological parameters in drinking water.

Bacterial toxicity/compatibility of platinum nanospheres, nanocuboids and nanoflowers

PubMed Central

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

2013-01-01

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

[Difference of three standard curves of real-time reverse-transcriptase PCR in viable Vibrio parahaemolyticus quantification].

PubMed

Jin, Mengtong; Sun, Wenshuo; Li, Qin; Sun, Xiaohong; Pan, Yingjie; Zhao, Yong

2014-04-04

We evaluated the difference of three standard curves in quantifying viable Vibrio parahaemolyticus in samples by real-time reverse-transcriptase PCR (Real-time RT-PCR). The standard curve A was established by 10-fold diluted cDNA. The cDNA was reverse transcripted after RNA synthesized in vitro. The standard curve B and C were established by 10-fold diluted cDNA. The cDNA was synthesized after RNA isolated from Vibrio parahaemolyticus in pure cultures (10(8) CFU/mL) and shrimp samples (10(6) CFU/g) (Standard curve A and C were proposed for the first time). Three standard curves were performed to quantitatively detect V. parahaemolyticus in six samples, respectively (Two pure cultured V. parahaemolyticus samples, two artificially contaminated cooked Litopenaeus vannamei samples and two artificially contaminated Litopenaeus vannamei samples). Then we evaluated the quantitative results of standard curve and the plate counting results and then analysed the differences. The three standard curves all show a strong linear relationship between the fractional cycle number and V. parahaemolyticus concentration (R2 > 0.99); The quantitative results of Real-time PCR were significantly (p < 0.05) lower than the results of plate counting. The relative errors compared with the results of plate counting ranked standard curve A (30.0%) > standard curve C (18.8%) > standard curve B (6.9%); The average differences between standard curve A and standard curve B and C were - 2.25 Lg CFU/mL and - 0.75 Lg CFU/mL, respectively, and the mean relative errors were 48.2% and 15.9%, respectively; The average difference between standard curve B and C was among (1.47 -1.53) Lg CFU/mL and the average relative errors were among 19.0% - 23.8%. Standard curve B could be applied to Real-time RT-PCR when quantify the number of viable microorganisms in samples.

Free Surface Effects on the Wake of a Flat Plate.

DTIC Science & Technology

1984-11-08

D-i46 98 FREE SURFCE’EFFECTS ON THE MAKE OF A FLAT PLTE(U) i/l 9(8 NAVAL RESEARCH LAB WASHINGTON DC T F SWEAN ET AL. 08 NOV 84 NRL-MR...5426UNCLASSIFIED F/ 20/4 NL 11111 ~ L.0 2 4 11111L .563 I -A 16 CEO -- . . IV NRL Memorandum Rpot52 Free Surface iEffwcs on the Wake of Al lit Plate T . F. SWEAlJ...13b. TIME COVERED 14. DATE OF REPORT (YeasrUonitDay) S.PAGE COUNT .0 - Interim IFROM _ TO T 1984 November 8 FS23 16 SUPPLEMENTARY NOTATION 17 COSATI

Quadrant anode image sensor

NASA Technical Reports Server (NTRS)

Lampton, M.; Malina, R. F.

1976-01-01

A position-sensitive event-counting electronic readout system for microchannel plates (MCPs) is described that offers the advantages of high spatial resolution and fast time resolution. The technique relies upon a four-quadrant electron-collecting anode located behind the output face of the microchannel plate, so that the electron cloud from each detected event is partly intercepted by each of the four quadrants. The relative amounts of charge collected by each quadrant depend on event position, permitting each event to be localized with two ratio circuits. A prototype quadrant anode system for ion, electron, and extreme ultraviolet imaging is described. The spatial resolution achieved, about 10 microns, allows individual MCP channels to be distinguished.

Effects of Gas Composition in the Modified Atmosphere Packaging on the Shelf-life of Longissimus dorsi of Korean Native Black Pigs-Duroc Crossbred during Refrigerated Storage

PubMed Central

Muhlisin; Panjono; Kim, Dong Soo; Song, Yeong Rae; Lee, Sung-Jin; Lee, Jeong Koo; Lee, Sung Ki

2014-01-01

This study was conducted to observe the effects of gas composition in modified atmosphere packaging (MAP) on the shelf-life of Longissimus dorsi of Korean Native Black Pigs-Duroc Crossbred (KNP×D) during refrigerated storage. Muscle sample was obtained from the left side of carcass of seven months old of KNP×D barrow. The sample was sliced into 1 cm in thickness, placed on trays (two slices/tray) and filled with different gas composition, i.e. 0:20:80/O2:CO2:N2 (MAP1), 30:20:50/O2:CO2:N2 (MAP2) and 70:20:10/O2:CO2:N2 (MAP3). Other slices of sample were vacuum packed (VP) as a control. All packs were stored at 5±1°C. At 12 d of storage, pH value of MAP2 and MAP3 were higher (p<0.05) than that of MAP1 and pH value of MAP1 was higher (p<0.05) than that of VP. At 6 d of storage, redness (a*) value of MAP2 and MAP3 were higher (p<0.05) than that of VP and MAP1 and, at 9 and 12 d of storage, redness value of MAP3 was higher (p<0.05) than that of VP, MAP1, and MAP2. At 3, 6, 9, and 12 d of storage, the 2-thiobarbituric acid reactive substances (TBARS) value of MAP3 was higher than that of MAP2 and TBARS value of MAP2 was higher than that of VP and MAP1. At 3, 6, 9, and 12 d of storage, volatile basic nitrogen values of MAP2 and MAP3 were higher (p<0.05) than those of VP and MAP1. At 3 d of storage, total aerobic plate counts of MAP2 and MAP3 were higher (p<0.05) than those of VP and MAP1 and, at 6 d of storage, total aerobic plate counts of MAP3 was higher (p<0.05) than that of MAP1 and MAP2. However, there was no significant different total aerobic plate count among MAP1, MAP2, and MAP3 at 9 and 12 d of storage. There was no significant different total anaerobic plate count among MAP1, MAP2, and MAP3 during storage. It is concluded that the MAP containing 30:20:50/O2:CO2:N2 gas composition (MAP2) might be ideal for better meat quality for KNP×D meat. PMID:25083110

New Method for Accurate Calibration of Micro-Channel Plate based Detection Systems and its use in the Fast Plasma Investigation of NASA's Magnetospheric MultiScale Mission

NASA Astrophysics Data System (ADS)

Gliese, U.; Avanov, L. A.; Barrie, A.; Kujawski, J. T.; Mariano, A. J.; Tucker, C. J.; Chornay, D. J.; Cao, N. T.; Zeuch, M.; Pollock, C. J.; Jacques, A. D.

2013-12-01

The Fast Plasma Investigation (FPI) of the NASA Magnetospheric MultiScale (MMS) mission employs 16 Dual Electron Spectrometers (DESs) and 16 Dual Ion Spectrometers (DISs) with 4 of each type on each of 4 spacecraft to enable fast (30ms for electrons; 150ms for ions) and spatially differentiated measurements of full the 3D particle velocity distributions. This approach presents a new and challenging aspect to the calibration and operation of these instruments on ground and in flight. The response uniformity and reliability of their calibration and the approach to handling any temporal evolution of these calibrated characteristics all assume enhanced importance in this application, where we attempt to understand the meaning of particle distributions within the ion and electron diffusion regions. Traditionally, the micro-channel plate (MCP) based detection systems for electrostatic particle spectrometers have been calibrated by setting a fixed detection threshold and, subsequently, measuring a detection system count rate plateau curve to determine the MCP voltage that ensures the count rate has reached a constant value independent of further variation in the MCP voltage. This is achieved when most of the MCP pulse height distribution (PHD) is located at higher values (larger pulses) than the detection amplifier threshold. This method is adequate in single-channel detection systems and in multi-channel detection systems with very low crosstalk between channels. However, in dense multi-channel systems, it can be inadequate. Furthermore, it fails to fully and individually characterize each of the fundamental parameters of the detection system. We present a new detection system calibration method that enables accurate and repeatable measurement and calibration of MCP gain, MCP efficiency, signal loss due to variation in gain and efficiency, crosstalk from effects both above and below the MCP, noise margin, and stability margin in one single measurement. The fundamental concepts of this method, named threshold scan, will be presented. It will be shown how to derive all the individual detection system parameters. This new method has been successfully applied to achieve a highly accurate calibration of the 16 Dual Electron Spectrometers and 16 Dual Ion Spectrometers of the MMS mission. The practical application of the method will be presented together with the achieved calibration results and their significance. Finally, it will be shown how this method will be applied to ensure the best possible in flight calibration during the mission.

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

NASA Astrophysics Data System (ADS)

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.

Development of Speckle Interferometry Algorithm and System

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

Shamsir, A. A. M.; Jafri, M. Z. M.; Lim, H. S.

2011-05-25

Electronic speckle pattern interferometry (ESPI) method is a wholefield, non destructive measurement method widely used in the industries such as detection of defects on metal bodies, detection of defects in intergrated circuits in digital electronics components and in the preservation of priceless artwork. In this research field, this method is widely used to develop algorithms and to develop a new laboratory setup for implementing the speckle pattern interferometry. In speckle interferometry, an optically rough test surface is illuminated with an expanded laser beam creating a laser speckle pattern in the space surrounding the illuminated region. The speckle pattern is opticallymore » mixed with a second coherent light field that is either another speckle pattern or a smooth light field. This produces an interferometric speckle pattern that will be detected by sensor to count the change of the speckle pattern due to force given. In this project, an experimental setup of ESPI is proposed to analyze a stainless steel plate using 632.8 nm (red) wavelength of lights.« less

Comparison of Candida Albicans Adherence to Conventional Acrylic Denture Base Materials and Injection Molding Acrylic Materials

PubMed Central

Aslanimehr, Masoomeh; Rezvani, Shirin; Mahmoudi, Ali; Moosavi, Najmeh

2017-01-01

Statement of the Problem: Candida species are believed to play an important role in initiation and progression of denture stomatitis. The type of the denture material also influences the adhesion of candida and development of stomatitis. Purpose: The aim of this study was comparing the adherence of candida albicans to the conventional and injection molding acrylic denture base materials. Materials and Method: Twenty injection molding and 20 conventional pressure pack acrylic discs (10×10×2 mm) were prepared according to their manufacturer’s instructions. Immediately before the study, samples were placed in sterile water for 3 days to remove residual monomers. The samples were then sterilized using an ultraviolet light unit for 10 minutes. 1×108 Cfu/ml suspension of candida albicans ATCC-10231 was prepared from 48 h cultured organism on sabouraud dextrose agar plates incubated at 37oC. 100 μL of this suspension was placed on the surface of each disk. After being incubated at 37oC for 1 hour, the samples were washed with normal saline to remove non-adherent cells. Attached cells were counted using the colony count method after shaking at 3000 rmp for 20 seconds. Finally, each group was tested for 108 times and the data were statistically analyzed by t-test. Results: Quantitative analysis revealed that differences in colony count average of candida albicans adherence to conventional acrylic materials (8.3×103) comparing to injection molding acrylic resins (6×103) were statistically significant (p<0.001). Conclusion: Significant reduction of candida albicans adherence to the injection acrylic resin materials makes them valuable for patients with high risk of denture stomatitis. PMID:28280761

Development of a flow cytometric method to analyze subpopulations of bacteria in probiotic products and dairy starters.

PubMed

Bunthof, Christine J; Abee, Tjakko

2002-06-01

Flow cytometry (FCM) is a rapid and sensitive technique that can determine cell numbers and measure various physiological characteristics of individual cells by using appropriate fluorescent probes. Previously, we developed an FCM assay with the viability probes carboxyfluorescein diacetate (cFDA) and TOTO-1 [1'-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)-bis-4-[3-methyl-2,3dihydro(benzo-1,3-oxazole)-2-methylidene]-1-(3'-trimethylammoniumpropyl)-pyridinium tetraiodide] for (stressed) lactic acid bacteria (C. J. Bunthof, K. Bloemen, P. Breeuwer, F. M. Rombouts, and T. Abee, Appl. Environ. Microbiol. 67:2326-2335, 2001). cFDA stains intact cells with enzymatic activity, and TOTO-1 stains membrane-permeabilized cells. Here we used this assay to study the viability of bacterial suspensions in milk, dairy fermentation starters, and probiotic products. To facilitate FCM analysis of bacteria in milk, a commercially available milk-clearing solution was used. The procedure was optimized to increase the signal-to-noise ratio. FCM enumerations were accurate down to a concentration of 10(5) cells ml(-1). The level of retrieval of Lactobacillus plantarum WCFS 1 suspended in milk was high, and viability was not affected by the procedure. The plate counts for cleared samples of untreated cell suspensions were nearly as high as the total FCM counts, and the correlation was strong (r > 0.99). In dairy fermentation starters and in probiotic products the FCM total cell counts were substantially higher than the numbers of CFU. Three functional populations could be distinguished: culturable cells, cells that are intact and metabolically active but not culturable, and permeabilized cells. The proportions of the populations differed in the products tested. This FCM method provides tools to assess the functionality of different populations in fermentation starters and probiotic products.

Development of a Flow Cytometric Method To Analyze Subpopulations of Bacteria in Probiotic Products and Dairy Starters

PubMed Central

Bunthof, Christine J.; Abee, Tjakko

2002-01-01

Flow cytometry (FCM) is a rapid and sensitive technique that can determine cell numbers and measure various physiological characteristics of individual cells by using appropriate fluorescent probes. Previously, we developed an FCM assay with the viability probes carboxyfluorescein diacetate (cFDA) and TOTO-1 {1′-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)-bis-4-[3-methyl-2,3dihydro(benzo-1,3-oxazole)-2-methylidene]-1-(3′-trimethylammoniumpropyl)-pyridinium tetraiodide} for (stressed) lactic acid bacteria (C. J. Bunthof, K. Bloemen, P. Breeuwer, F. M. Rombouts, and T. Abee, Appl. Environ. Microbiol. 67:2326-2335, 2001). cFDA stains intact cells with enzymatic activity, and TOTO-1 stains membrane-permeabilized cells. Here we used this assay to study the viability of bacterial suspensions in milk, dairy fermentation starters, and probiotic products. To facilitate FCM analysis of bacteria in milk, a commercially available milk-clearing solution was used. The procedure was optimized to increase the signal-to-noise ratio. FCM enumerations were accurate down to a concentration of 105 cells ml−1. The level of retrieval of Lactobacillus plantarum WCFS 1 suspended in milk was high, and viability was not affected by the procedure. The plate counts for cleared samples of untreated cell suspensions were nearly as high as the total FCM counts, and the correlation was strong (r > 0.99). In dairy fermentation starters and in probiotic products the FCM total cell counts were substantially higher than the numbers of CFU. Three functional populations could be distinguished: culturable cells, cells that are intact and metabolically active but not culturable, and permeabilized cells. The proportions of the populations differed in the products tested. This FCM method provides tools to assess the functionality of different populations in fermentation starters and probiotic products. PMID:12039752

Microplate Fluorescence Assay for Measurement of the Ability of Strains of Listeria monocytogenes from Meat and Meat-Processing Plants To Adhere to Abiotic Surfaces▿

PubMed Central

Gamble, Rachel; Muriana, Peter M.

2007-01-01

Listeria monocytogenes is a significant food-borne pathogen that is capable of adhering to and producing biofilms on processing equipment, making it difficult to eliminate from meat-processing environments and allowing potential contamination of ready-to-eat (RTE) products. We devised a fluorescence-based microplate method for screening isolates of L. monocytogenes for the ability to adhere to abiotic surfaces. Strains of L. monocytogenes were incubated for 2 days at 30°C in 96-well microplates, and the plates were washed in a plate washer. The retained cells were incubated for 15 min at 25°C with 5,6-carboxyfluorescein diacetate and washed again, and then the fluorescence was read with a plate reader. Several enzymatic treatments (protease, lipase, and cellulase) were effective in releasing adherent cells from the microplates, and this process was used for quantitation on microbiological media. Strongly adherent strains of L. monocytogenes were identified that had 15,000-fold-higher levels of fluorescence and 100,000-fold-higher plate counts in attachment assays than weakly adherent strains. Strongly adherent strains of L. monocytogenes adhered equally well to four different substrates (glass, plastic, rubber, and stainless steel); showed high-level attachment on microplates at 10, 20, 30, and 40°C; and showed significant differences from weakly adherent strains when examined by scanning electron microscopy. A greater incidence of strong adherence was observed for strains isolated from RTE meats than for those isolated from environmental surfaces. Analysis of surface adherence among Listeria isolates from processing environments may provide a better understanding of the molecular mechanisms involved in attachment and suggest solutions to eliminate them from food-processing environments. PMID:17586676

Effects of dark storage and retail display on beef chuck and round muscles enhanced with ammonium hydroxide, salt, and carbon monoxide.

PubMed

Hamling, A E; Jenschke, B E; Calkins, C R

2008-04-01

The objective of this study was to determine the retail shelf stability of beef chuck and round muscles enhanced with ammonium hydroxide, salt, and carbon monoxide. A split plot design was used for each of 3 muscles [triceps brachii (TB), biceps femoris (BF), and rectus femoris (RF)] with 2 treatments (0 and 20% pump), 3 dark storage periods (1, 2, and 3 wk), and 3 replications in the whole plot and retail display period as the split plot. There were a total of 12 subprimals per treatment per dark storage period (n = 72 each). Individual steaks were cut to a thickness of 2.54 cm and packaged in a modified-atmosphere package (MAP). The TB was packaged in a high-oxygen MAP (80% oxygen, 20% carbon dioxide). The BF and RF were packaged in a low-oxygen MAP (100% carbon dioxide). At the completion of each dark storage period, steaks were subjected to 7 d of simulated retail display. Steaks were used for objective and subjective color measurements, total plate counts, and determination of retail purge and oxidation. For all muscles, total plate counts were always numerically greater in injected steaks. Triceps brachii steaks held in dark storage for 3 wk and displayed at retail for 4 or more days all exceeded 10(7) log of cfu/cm(2) for aerobic plate count. Biceps femoris and RF steaks packaged in a low-oxygen MAP had much lower bacterial counts, with levels below 4.2 log of cfu/cm(2), even after 7 d of retail display. Oxidation values for the TB were extremely high (ranging from 12.3 to 26.6), whereas the BF and RF had values that were much lower (< or =1.0 mg of malonaldehyde/kg of muscle), likely due to the oxidation occurring in a high-oxygen MAP for the TB. Enhanced TB steaks proved to have greater color stability (less discoloration) than nonenhanced TB steaks. In addition, the BF and RF (low-oxygen MAP) steaks had better color stability (more stable redness values) than TB (high-oxygen MAP) steaks, although TB steaks initially exhibited a brighter red color. Retail display life was enhanced by packaging in 100% carbon dioxide, and enhanced steaks exhibited greater color stability in retail display than control steaks.

7 CFR 58.648 - Microbiological requirements for ice cream.

Code of Federal Regulations, 2012 CFR

2012-01-01

... requirements for ice cream. The finished product shall contain not more than 50,000 bacteria per gram as determined by the standard plate count, and shall contain not more than 10 coliform organisms per gram for plain and not more than 20 coliform per gram in chocolate, fruit, nut or other flavors in three out of...

7 CFR 58.648 - Microbiological requirements for ice cream.

Code of Federal Regulations, 2013 CFR

2013-01-01

... requirements for ice cream. The finished product shall contain not more than 50,000 bacteria per gram as determined by the standard plate count, and shall contain not more than 10 coliform organisms per gram for plain and not more than 20 coliform per gram in chocolate, fruit, nut or other flavors in three out of...

7 CFR 58.648 - Microbiological requirements for ice cream.

Code of Federal Regulations, 2011 CFR

2011-01-01

... requirements for ice cream. The finished product shall contain not more than 50,000 bacteria per gram as determined by the standard plate count, and shall contain not more than 10 coliform organisms per gram for plain and not more than 20 coliform per gram in chocolate, fruit, nut or other flavors in three out of...

7 CFR 58.648 - Microbiological requirements for ice cream.

Code of Federal Regulations, 2014 CFR

2014-01-01

... requirements for ice cream. The finished product shall contain not more than 50,000 bacteria per gram as determined by the standard plate count, and shall contain not more than 10 coliform organisms per gram for plain and not more than 20 coliform per gram in chocolate, fruit, nut or other flavors in three out of...

7 CFR 58.648 - Microbiological requirements for ice cream.

Code of Federal Regulations, 2010 CFR

2010-01-01

... requirements for ice cream. The finished product shall contain not more than 50,000 bacteria per gram as determined by the standard plate count, and shall contain not more than 10 coliform organisms per gram for plain and not more than 20 coliform per gram in chocolate, fruit, nut or other flavors in three out of...

Is hand hygiene before putting on nonsterile gloves in the intensive care unit a waste of health care worker time?--a randomized controlled trial.

PubMed

Rock, Clare; Harris, Anthony D; Reich, Nicholas G; Johnson, J Kristie; Thom, Kerri A

2013-11-01

Hand hygiene (HH) is recognized as a basic effective measure in prevention of nosocomial infections. However, the importance of HH before donning nonsterile gloves is unknown, and few published studies address this issue. Despite the lack of evidence, the World Health Organization and other leading bodies recommend this practice. The aim of this study was to assess the utility of HH before donning nonsterile gloves prior to patient contact. A prospective, randomized, controlled trial of health care workers entering Contact Isolation rooms in intensive care units was performed. Baseline finger and palm prints were made from dominant hands onto agar plates. Health care workers were then randomized to directly don nonsterile gloves or perform HH and then don nonsterile gloves. Postgloving finger and palm prints were then made from the gloved hands. Plates were incubated and colony-forming units (CFU) of bacteria were counted. Total bacterial colony counts of gloved hands did not differ between the 2 groups (6.9 vs 8.1 CFU, respectively, P = .52). Staphylococcus aureus was identified from gloves (once in "hand hygiene prior to gloving" group, twice in "direct gloving" group). All other organisms were expected commensal flora. HH before donning nonsterile gloves does not decrease already low bacterial counts on gloves. The utility of HH before donning nonsterile gloves may be unnecessary. Copyright © 2013 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

Microbiological quality and antibiotic residues in informally marketed raw cow milk within the coastal savannah zone of Ghana.

PubMed

Addo, K K; Mensah, G I; Aning, K G; Nartey, N; Nipah, G K; Bonsu, C; Akyeh, M L; Smits, H L

2011-02-01

To investigate the microbiological quality and the presence of antibiotic residues in raw cow milk and in some indigenous milk products produced and marketed by the informal sector in the coastal savannah zone of Ghana. Milk samples were aseptically collected from 224 kraals and samples of 26 indigenous milk products were purchased from processors and retailers. Total plate counts, total coliform counts and the presence of Escherichia coli and E. coli O157:H7 were determined in all 250 samples. Milk samples were also tested for antibiotic residues. Total plate counts exceeded 10⁵ CFU/ml in 45.2% of the samples while coliforms exceeded 10³ CFU/ml in 66.0% and E. coli was detected in 11.2%. E. coli was present in raw cow milk but not in the indigenous products and all E. coli isolates were negative for E. coli O157:H7. Antibiotic residues were detected in 3.1% of the raw cow milk samples. Bulk milk contains unacceptable levels of hygiene indicators and antibiotic residues and is a potential source of milk-borne infections. The detection of E. coli and antibiotic residues raises public health concerns about the safety of fresh unpasteurized cow milk in the coastal savannah zone of Ghana and calls for improved farm hygiene, the need for milk pasteurization and the sensible use of antibiotics in the milk industry. © 2010 Blackwell Publishing Ltd.

The use of detergents and sanitizers in dairy farm sanitation--an updated perspective.

PubMed

Gilbert, P H

1982-06-01

Raw milk quality in South Africa is poor and standard plate counts in the millions per ml are common. This is largely due to inefficient cleaning and sanitizing of dairy equipment. The basic constituents in milk are described and various soils are classified as soluble in water, alkali, acid, solvent or surfactant or as insoluble. The importance of water quality is highlighted and the influence of mineral salts on soil deposition described. Dairy detergents are broadly classified as alkaline or acid, the former being most effective against fatty and proteinaceous soils and the latter effective against mineral salts. Typical detergent ingredients and their properties are described. Chlorine is incorporated into alkaline detergents not as a sanitizing agent, but as a peptizing agent to aid in protein soil removal. At high pH values the antimicrobial activity of chlorine is greatly diminished. The use of a daily acidified rinse (pH 3,0-5,0) is preferred to the periodic acid wash, since the acid rinse prevents mineral deposition rather than removing accumulated milkstone. All cleaning programmes follow the same fundamental steps--Pre-rinse (40-50 degrees C), wash (60-70 degrees C), rinse (pH 3,0-5,0) and sanitize (25 ppm iodine and 100 ppm chlorine). Farms following such a programme are able to achieve Standard Plate Counts of less than 10,000/ml and coliform counts of less than 10/ml for raw milk.

Avian leucocyte counting using the hemocytometer

USGS Publications Warehouse

Dein, F.J.; Wilson, A.; Fischer, D.; Langenberg, P.

1994-01-01

Automated methods for counting leucocytes in avian blood are not available because of the presence of nucleated erythrocytes and thrombocytes. Therefore, total white blood cell counts are performed by hand using a hemocytometer. The Natt and Herrick and the Unopette methods are the most common stain and diluent preparations for this procedure. Replicate hemocytometer counts using these two methods were performed on blood from four birds of different species. Cells present in each square of the hemocytometer were counted. Counting cells in the corner, side, or center hemocytometer squares produced statistically equivalent results; counting four squares per chamber provided a result similar to that obtained by counting nine squares; and the Unopette method was more precise for hemocytometer counting than was the Natt and Herrick method. The Unopette method is easier to learn and perform but is an indirect process, utilizing the differential count from a stained smear. The Natt and Herrick method is a direct total count, but cell identification is more difficult.

Assessing the Accuracy of a Wrist Motion Tracking Method for Counting Bites across Demographic and Food Variables

PubMed Central

Shen, Yiru; Salley, James; Muth, Eric; Hoover, Adam

2017-01-01

This paper describes a study to test the accuracy of a method that tracks wrist motion during eating to detect and count bites. The purpose was to assess its accuracy across demographic (age, gender, ethnicity) and bite (utensil, container, hand used, food type) variables. Data were collected in a cafeteria under normal eating conditions. A total of 271 participants ate a single meal while wearing a watch-like device to track their wrist motion. Video was simultaneously recorded of each participant and subsequently reviewed to determine the ground truth times of bites. Bite times were operationally defined as the moment when food or beverage was placed into the mouth. Food and beverage choices were not scripted or restricted. Participants were seated in groups of 2–4 and were encouraged to eat naturally. A total of 24,088 bites of 374 different food and beverage items were consumed. Overall the method for automatically detecting bites had a sensitivity of 75% with a positive predictive value of 89%. A range of 62–86% sensitivity was found across demographic variables, with slower eating rates trending towards higher sensitivity. Variations in sensitivity due to food type showed a modest correlation with the total wrist motion during the bite, possibly due to an increase in head-towards-plate motion and decrease in hand-towards-mouth motion for some food types. Overall, the findings provide the largest evidence to date that the method produces a reliable automated measure of intake during unrestricted eating. PMID:28113994

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

PubMed

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.

Mode I Failure of Armor Ceramics: Experiments and Modeling

NASA Astrophysics Data System (ADS)

Meredith, Christopher; Leavy, Brian

2017-06-01

The pre-notched edge on impact (EOI) experiment is a technique for benchmarking the damage and fracture of ceramics subjected to projectile impact. A cylindrical projectile impacts the edge of a thin rectangular plate with a pre-notch on the opposite edge. Tension is generated at the notch tip resulting in the initiation and propagation of a mode I crack back toward the impact edge. The crack can be quantitatively measured using an optical method called Digital Gradient Sensing, which measures the crack-tip deformation by simultaneously quantifying two orthogonal surface slopes via measuring small deflections of light rays from a specularly reflective surface around the crack. The deflections in ceramics are small so the high speed camera needs to have a very high pixel count. This work reports on the results from pre-crack EOI experiments of SiC and B4 C plates. The experimental data are quantitatively compared to impact simulations using an advanced continuum damage model. The Kayenta ceramic model in Alegra will be used to compare fracture propagation speeds, bifurcations and inhomogeneous initiation of failure will be compared. This will provide insight into the driving mechanisms required for the macroscale failure modeling of ceramics.

A valid and reliable method to measure jump-specific training and competition load in elite volleyball players.

PubMed

Skazalski, C; Whiteley, R; Hansen, C; Bahr, R

2018-05-01

Use of a commercially available wearable device to monitor jump load with elite volleyball players has become common practice. The purpose of this study was to evaluate the validity and reliability of this device, the Vert, to count jumps and measure jump height with professional volleyball players. Jump count accuracy was determined by comparing jumps recorded by the device to jumps observed through systematic video analysis of three practice sessions and two league matches performed by a men's professional volleyball team. Jumps performed by 14 players were each coded for time and jump type and individually matched to device recorded jumps. Jump height validity of the device was examined against reference standards as participants performed countermovement jumps on a force plate and volleyball-specific jumps with a Vertec. The Vert device accurately counted 99.3% of the 3637 jumps performed during practice and match play. The device showed excellent jump height interdevice reliability for two devices placed in the same pouch during volleyball jumps (r = .99, 95% CI 0.98-0.99). The device had a minimum detectable change (MDC) of 9.7 cm and overestimated jump height by an average of 5.5 cm (95% CI 4.5-6.5) across all volleyball jumps. The Vert device demonstrates excellent accuracy counting volleyball-specific jumps during training and competition. While the device is not recommended to measure maximal jumping ability when precision is needed, it provides an acceptable measure of on-court jump height that can be used to monitor athlete jump load. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

High throughput RNAi assay optimization using adherent cell cytometry.

PubMed

Nabzdyk, Christoph S; Chun, Maggie; Pradhan, Leena; Logerfo, Frank W

2011-04-25

siRNA technology is a promising tool for gene therapy of vascular disease. Due to the multitude of reagents and cell types, RNAi experiment optimization can be time-consuming. In this study adherent cell cytometry was used to rapidly optimize siRNA transfection in human aortic vascular smooth muscle cells (AoSMC). AoSMC were seeded at a density of 3000-8000 cells/well of a 96 well plate. 24 hours later AoSMC were transfected with either non-targeting unlabeled siRNA (50 nM), or non-targeting labeled siRNA, siGLO Red (5 or 50 nM) using no transfection reagent, HiPerfect or Lipofectamine RNAiMax. For counting cells, Hoechst nuclei stain or Cell Tracker green were used. For data analysis an adherent cell cytometer, Celigo® was used. Data was normalized to the transfection reagent alone group and expressed as red pixel count/cell. After 24 hours, none of the transfection conditions led to cell loss. Red fluorescence counts were normalized to the AoSMC count. RNAiMax was more potent compared to HiPerfect or no transfection reagent at 5 nM siGLO Red (4.12 +/-1.04 vs. 0.70 +/-0.26 vs. 0.15 +/-0.13 red pixel/cell) and 50 nM siGLO Red (6.49 +/-1.81 vs. 2.52 +/-0.67 vs. 0.34 +/-0.19). Fluorescence expression results supported gene knockdown achieved by using MARCKS targeting siRNA in AoSMCs. This study underscores that RNAi delivery depends heavily on the choice of delivery method. Adherent cell cytometry can be used as a high throughput-screening tool for the optimization of RNAi assays. This technology can accelerate in vitro cell assays and thus save costs.

Addition of DNase Improves the In Vitro Activity of Antifungal Drugs against Candida albicans Biofilms

PubMed Central

Martins, Margarida; Henriques, Mariana; Lopez-Ribot, José L.; Oliveira, Rosário

2011-01-01

SUMMARY Background Cells within Candida albicans biofilms display decreased susceptibility to most clinically used antifungal agents. We recently demonstrated that extracellular DNA (eDNA) plays an important role in biofilm integrity, as a component of the biofilm matrix. Objective To gain insight into the contributions of eDNA to C. albicans biofilms antifungal susceptibility by the investigation of the impact of the combined use of deoxyribonuclease I (DNase) and antifungals to treat biofilms. Methods C. albicans biofilms were formed using a simple and reproducible 96-well plate-based method. The activity of the combined use of 0.13 mg l−1 DNase and antifungals was estimated by the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide (XTT) reduction assay, and total viable counts. Results and Conclusions Here we report the improved efficacy of amphotericin B when in combination with DNase against C. albicans biofilms, as assessed by XTT readings and viable counts. Furthermore, although DNase increased the efficacy of caspofungin in the reduction of mitochondrial activity, no changes were observed in terms of culturable cells. DNase did not affect biofilm cells susceptibility to fluconazole. This work suggests that agents that target processes affecting the biofilm structural integrity may have potential use as adjuvants of a catheter–lock therapy. PMID:21668524

Study on biogenic amines in various dry salted fish consumed in China

NASA Astrophysics Data System (ADS)

Wu, Yanyan; Chen, Yufeng; Li, Laihao; Yang, Xianqing; Yang, Shaoling; Lin, Wanling; Zhao, Yongqiang; Deng, Jianchao

2016-08-01

This study was carried out to investigate the biogenic amines (BAs), physicochemical property and microorganisms in dry salted fish, a traditional aquatic food consumed in China. Forty three samples of dry salted fish were gathered from retail and wholesale markets and manufacturers, which had been produced in various regions in China. Cadaverine (CAD) and putrescine (PUT) were quantitatively the most common biogenic amines. About 14% of the samples exceeded the histamine content standards established by the FDA and/or EU. The highest histamine content was found in Silver pomfret ( Pampus argenteus) (347.79 mg kg-1). Five of forty three samples exceeded the acceptable content of TYR (100 mg kg-1), and 23.26% of dried-salted fish contained high contents of biogenic amines (above 600 mg kg-1). In addition, species, regions, pickling processes and drying methods made the physicochemical property, microorganisms and biogenic amines in dry salted fish to be different to some extents. The total plate count (TPC) was much higher than that of total halophilic bacteria in all samples. The biogenic amines, physicochemical property and microbiological counts exhibited large variations among samples. Furthermore, no significant correlation between biogenic amines and physicochemical property and TPC was observed. This study indicated that dry salted fish may still present healthy risk for BAs, depending on the processing methods, storage conditions among others.

Effect of Iranian Ziziphus honey on growth of some foodborne pathogens

PubMed Central

Ekhtelat, Maryam; Ravaji, Karim; Parvari, Masood

2016-01-01

Background: Honey has previously been shown to have wound healing and antimicrobial properties, but this is dependent on the type of honey, geographical location, and flower from which the final product is derived. We tested the antimicrobial activity of a natural honey originating from the Ziziphus spina-christi tree, against selected strains of bacteria. Ziziphus honey among more than a 100 verities of honey is known to have the greatest value of energy and minerals in it. Materials and Methods: The aim of this study was to determine the antibacterial activity of Ziziphus honey in 10%, 20%, 30%, and 40% dilutions (v/v) against Listeria monocytogenes, Salmonella typhimurium, Escherichia coli, and Staphylococcus aureus. Viable count enumeration of the sample was investigated after 0, 24, 72, and 120 h postinoculation with any of the bacteria using pour-plate method. Results: The findings indicate that Ziziphus honey was effective against these pathogenic bacteria. In a comparative trial, antibacterial activity of Ziziphus honey was higher after 120 h incubation for each four bacteria in most dilutions. The microbial count showed 3-7.5 log reduction comparing with control after 120 h. Conclusions: Therefore, it is recommended using Ziziphus honey as a natural preservative and antibacterial agent. Also, it could potentially be used as therapeutic agents against bacterial infection particularly to the tested microorganisms. PMID:27003970

Treatment of Palm Oil Mill Effluent by a Microbial Consortium Developed from Compost Soils

PubMed Central

Nwuche, Charles O.; Ogbonna, James C.

2014-01-01

A method for the aerobic treatment of palm oil mill effluent (POME) was investigated in shake-flask experiments using a consortium developed from POME compost. POME was initially centrifuged at 4,000 g for 15 min and the supernatant was enriched with (NH4)2SO4 (0.5%) and yeast extract (0.25%) to boost its nitrogen content. At optimum pH (pH 4) and temperature (40°C) conditions, the chemical oxygen demand (COD) of the effluent decreased from 10,350 to 1,000 mg/L (90.3%) after 7 days. The total bacterial population determined by plate count enumeration was 2.4 × 106 CFU/mL, while the fungal count was 1.8 × 103 colonies/mL. Bacteria of the genera Pseudomonas, Flavobacterium, Micrococcus, and Bacillus were isolated, while the fungal genera included Aspergillus, Penicillium, Trichoderma, and Mucor. When the isolated species were each inoculated into separate batches of the raw effluent, both pH and COD were unchanged. However, at 75 and 50% POME dilutions, the COD dropped by 52 and 44%, respectively, while the pH increased from 4 to 7.53. POME treatment by aerobic method is sustainable and holds promising prospects for cushioning the environment from the problems associated with the use of anaerobic systems. PMID:27433536

Efficacy of High-volume Evacuator in Aerosol Reduction: Truth or Myth? A Clinical and Microbiological Study.

PubMed

Desarda, Hitesh; Gurav, Abhijit; Dharmadhikari, Chandrakant; Shete, Abhijeet; Gaikwad, Subodh

2014-01-01

Background and aims. Basic periodontal treatment aims at eliminating supra- and sub-gingival plaque and establishing conditions which will allow effective self-performed plaque control. This aim is primarily achieved with sonic and ultrasonic scalers. However, generation of bacterial aerosols during these procedures is of great concern to patients, the dentist and the dental assistant. The aim of this study was to compare the reduction in aerosol with and without high-volume evacuator through a microbiological study. Materials and methods. For this clinical study a fumigated closed operatory was selected. Maxillary incisors and canines were selected as an area for scaling. Piezoelectric ultrasonic scaling was performed in the absence and in the presence of a high-volume evacuator at 12 and 20 inches from the patient's oral cavity. In both groups scaling was carried out for 10 minutes. Nutrient agar plates were exposed for a total of 20 minutes. After this procedure, nutrient agar plates were incubated in an incubator at 37°C for 24 hours. The next day the nutrient agar plates were examined for colony forming units by a single microbiologist. Results. The results showed no statistically significant differences in colony forming units (CFU) with and without the use of a high-volume evacuator either at 12 or 20 inches from the patient's oral cavity. Conclusion. It was concluded that high-volume evacuator, when used as a separate unit without any modification, is not effective in reducing aerosol counts and environmental contamination.

The Effect of Estradiol on the Growth Plate Chondrocytes of Limb and Spine from Postnatal Mice in vitro: The Role of Estrogen-Receptor and Estradiol Concentration.

PubMed

Shi, Sheng; Zheng, Shuang; Li, Xin-Feng; Liu, Zu-De

2017-01-01

Objectives: Skeletal development is a complex process. Little is known about the different response of limb or spine growth plate chondrocytes (LGP or SGP) to the estrogen level and the role of estrogen receptor (ER) during postnatal stage. Methods: LGP and SGP chondrocytes were isolated from 50 one-week mice and treated with different concentrations of 17β-estradiol. Cell viability was measured by cell counting kit-8 (CCK-8). The expression of collagen II and X were evaluated by real-time PCR and Western blotting. Then, the response of LGP or SGP chondrocyte after with or without estradiol and specific ER antagonists to block the effect of ERs were also measured by Western blotting and immunofluorescence. Results: Estradiol promoted the chondrogensis of the chondrocytes in vitro and achieved the maximal expression of type II collagen at the dose of 10 -7 M. Additionally, the regulatory effect of estradiol on the chondrogenesis can be mainly relied on ERα. The LGP chondrocytes were more sensitive to the estradiol treatment than SGP in the expression of type II collagen. Conclusions: Estrogen at a pharmacological concentration (10 -7 M) could stimulate the maximal production of type II collagen in the growth plate chondrocytes in vitro, which exerts its activity mainly through ERα in the chondrogenesis. Furthermore, the LGP chondrocytes were more sensitive to the estradiol treatment than SGP in the chondrogenesis.

Study of the microbial ecology of wild and aquacultured Tunisian fresh fish.

PubMed

Boulares, Mouna; Mejri, Lobna; Hassouna, Mnasser

2011-10-01

Eighty samples of fresh fish were collected in Tunisia and analyzed for microbial load. Quality and hygienic safety of the meat and intestines of wild and aquacultured fresh fish were determined. The mesophilic aerobic plate count and populations of psychrotrophic lactic acid bacteria (LAB) and other psychrotrophic bacteria ranged from 5.67 to 7.29, 4.51 to 6, and 5.07 to 6.21 log CFU/g, respectively. For all microbiological determinations, bacterial counts were lower in meat than in the intestines of fresh fish. For all samples lower microbial populations were found in most of the wild fish than in the aquacultured fish. No isolates of the pathogenic genera Salmonella and Listeria were detected in any sample. Among the 160 strains of biopreservative psychrotrophic LAB and the 150 strains of spoilage psychrotrophic gram-negative bacteria identified by biochemical and molecular methods, Lactobacillus (six species) and Pseudomonas (six species) predominated. Lactococcus, Leuconostoc, Carnobacterium (C. piscicola and C. divergens), Aeromonas, and Photobacterium were the most common genera, and Lactococcus lactis, Lactobacillus plantarum, Pseudomonas fluorescens, and Aeromonas hydrophila were the most common species. These findings indicate that the microbiological quality of fresh fish in Tunisia can be preserved by controlling pathogenic and psychrotrophic bacteria.

Study on spoilage capability and VBNC state formation and recovery of Lactobacillus plantarum.

PubMed

Liu, Junyan; Li, Lin; Li, Bing; Peters, Brian M; Deng, Yang; Xu, Zhenbo; Shirtliff, Mark E

2017-09-01

The present study aimed at investigating the capability of L. plantarum strain BM-LP14723 to enter into and recover from the viable but nonculturable (VBNC) state and to cause beer spoilage. VBNC state was induced by incubating in beer with subculturing or low temperature treatment. Culturable, total, and viable cells numbers were assessed by MRS agar plate counting, acridine orange direct counting, and Live/Dead BacLight bacterial viability kit, respectively. Organic acids concentrations were measured by reversed-phase high performance liquid chromatography. VBNC L. plantarum cells were detected after 189 ± 1.9 days low temperature treatment or 29 ± 0.7 subcultures in beer. The VBNC L. plantarum retained spoilage capability. Addition of catalase is an effective method for the recovery of the VBNC L. plantarum cells. L. plantarum strain BM-LP14723 is capable of entering into and recovery from the VBNC state and maintained spoilage capability. The current study presented that beer-spoilage L. plantarum can hide both in breweries and during transporting and marketing process and thus lead to beer-spoilage incidents. Copyright © 2017 Elsevier Ltd. All rights reserved.

New methods for the detection of viruses: call for review of drinking water quality guidelines.

PubMed

Grabow, W O; Taylor, M B; de Villiers, J C

2001-01-01

Drinking water supplies which meet international recommendations for source, treatment and disinfection were analysed. Viruses recovered from 100 L-1,000 L volumes by in-line glass wool filters were inoculated in parallel into four cell culture systems. Cell culture inoculation was used to isolate cytopathogenic viruses, amplify the nucleic acid of non-cytopathogenic viruses and confirm viability of viruses. Over a period of two years, viruses were detected in 23% of 413 drinking water samples and 73% of 224 raw water samples. Cytopathogenic viruses were detected in 6% raw water samples but not in any treated drinking water supplies. Enteroviruses were detected in 17% drinking water samples, adenoviruses in 4% and hepatitis A virus in 3%. In addition to these viruses, astro- and rotaviruses were detected in raw water. All drinking water supplies had heterotrophic plate counts of < 100/mL, total and faecal coliform counts of 0/100 mL and negative results in qualitative presence-absence tests for somatic and F-RNA coliphages (500 mL samples). These results call for a revision of water quality guidelines based on indicator organisms and vague reference to the absence of viruses.

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

PubMed Central

Al-Kharousi, Zahra S.; 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 biochemical methods (VITEK 2). Enterobacteriaceae occurred in 60% of fruits and 91% of vegetables. Enterococcus was isolated from 20% of fruits and 42% of vegetables. E. coli and S. aureus were isolated from 22% and 7% of vegetables, respectively. Ninety-seven bacteria comprising 21 species were similarly identified by VITEK 2 and PCR to species level. E. coli, Klebsiella pneumoniae, Enterococcus casseliflavus, and Enterobacter cloacae were the most abundant species; many are known as opportunistic pathogens which may raise concern to improve the microbial quality of fresh produce. Phylogenetic trees showed no relationship between clustering of the isolates based on the 16S rRNA gene and the original countries of fresh produce. Intercountry passage of opportunistic pathogens in fresh produce cannot be ruled out, which requires better management. PMID:26989419

Antibacterial activity of antibacterial cutting boards in household kitchens.

PubMed

Kounosu, Masayuki; Kaneko, Seiichi

2007-12-01

We examined antibacterial cutting boards with antibacterial activity values of either "2" or "4" in compliance with the JIS Z 2801 standard, and compared their findings with those of cutting boards with no antibacterial activity. These cutting boards were used in ten different households, and we measured changes in the viable cell counts of several types of bacteria with the drop plate method. We also identified the detected bacterial flora and measured the minimum antimicrobial concentrations of several commonly used antibacterial agents against the kinds of bacteria identified to determine the expected antibacterial activity of the respective agents. Cutting boards with activity values of both "2" and "4" proved to be antibacterial in actual use, although no correlation between the viable cell counts and the antibacterial activity values was observed. In the kitchen environment, large quantities of Pseudomonas, Flavobacterium, Micrococcus, and Bacillus were detected, and it was confirmed that common antibacterial agents used in many antibacterial products are effective against these bacterial species. In addition, we measured the minimum antimicrobial concentrations of the agents against lactobacillus, a typical good bacterium, and discovered that this bacterium is less sensitive to these antibacterial agents compared to more common bacteria.

Analysis of Acoustic Emission Parameters from Corrosion of AST Bottom Plate in Field Testing

NASA Astrophysics Data System (ADS)

Jomdecha, C.; Jirarungsatian, C.; Suwansin, W.

Field testing of aboveground storage tank (AST) to monitor corrosion of the bottom plate is presented in this chapter. AE testing data of the ten AST with different sizes, materials, and products were employed to monitor the bottom plate condition. AE sensors of 30 and 150 kHz were used to monitor the corrosion activity of up to 24 channels including guard sensors. Acoustic emission (AE) parameters were analyzed to explore the AE parameter patterns of occurring corrosion compared to the laboratory results. Amplitude, count, duration, and energy were main parameters of analysis. Pattern recognition technique with statistical was implemented to eliminate the electrical and environmental noises. The results showed the specific AE patterns of corrosion activities related to the empirical results. In addition, plane algorithm was utilized to locate the significant AE events from corrosion. Both results of parameter patterns and AE event locations can be used to interpret and locate the corrosion activities. Finally, basic statistical grading technique was used to evaluate the bottom plate condition of the AST.

Preliminary investigation of a water-based method for fast integrating mobility spectrometry

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

Spielman, Steven R.; Hering, Susanne V.; Kuang, Chongai

A water-based condensational growth channel was developed for imaging mobility-separated particles within a parallel plate separation channel of the Fast Integrated Mobility Spectrometer (FIMS). Reported are initial tests of that system, in which the alcohol condenser of the FIMS was replaced by a water-based condensational growth channel. Tests with monodispersed sodium chloride aerosol verify that the water-condensational growth maintained the laminar flow, while providing sufficient growth for particle imaging. Particle positions mapped onto particle mobility, in accordance with theoretical expectations. Particles ranging in size from 12 nm to 100 nm were counted with the same efficiency as with a butanol-based ultrafine particlemore » counter, once inlet and line losses were taken into account.« less

Data on microbial and physiochemical characteristics of inlet and outlet water from household water treatment devices in Rasht, Iran.

PubMed

Naghipour, Dariush; Ashrafi, Seyed Davoud; Mojtahedi, Ali; Vatandoost, Masoud; Hosseinzadeh, Loghman; Roohbakhsh, Esmail

2018-02-01

In this research, we measured various parameters related to drinking water quality include turbidity, temperature, pH, EC, TDS, Alkalinity, fecal and total coliform, heterotrophic plate count (HPC), free chlorine, Mn, Ca, Mg, Fe, Na, Cl - , F - , HCO 3 , in the inlet and outlet of household water treatment devices according to the standard methods for the examination of water and wastewater (W.E. Federation and Association and A.P.H., 2005) [1]. Sixty four inlet and outlet water samples were taken from thirty two household water treatment devices from eight different residential blocks in Golsar town of Rasht, Iran. The data obtained from experiments were analyzed using the software Special Package for Social Sciences (SPSS 24) and MS-Excel.

Preliminary investigation of a water-based method for fast integrating mobility spectrometry

DOE PAGES

Spielman, Steven R.; Hering, Susanne V.; Kuang, Chongai; ...

2017-06-06

A water-based condensational growth channel was developed for imaging mobility-separated particles within a parallel plate separation channel of the Fast Integrated Mobility Spectrometer (FIMS). Reported are initial tests of that system, in which the alcohol condenser of the FIMS was replaced by a water-based condensational growth channel. Tests with monodispersed sodium chloride aerosol verify that the water-condensational growth maintained the laminar flow, while providing sufficient growth for particle imaging. Particle positions mapped onto particle mobility, in accordance with theoretical expectations. Particles ranging in size from 12 nm to 100 nm were counted with the same efficiency as with a butanol-based ultrafine particlemore » counter, once inlet and line losses were taken into account.« less

Microbial loads and antibiotic resistance patterns of Staphylococcus aureus in different types of raw poultry-based meat preparations.

PubMed

Buzón-Durán, Laura; Capita, Rosa; Alonso-Calleja, Carlos

2017-09-01

The hygiene status of raw chicken-meat preparations from retail outlets in North-Western Spain was investigated. Microbial counts (aerobic plate counts (APCs), psychrotrophs, Enterobacteriaceae, fecal coliforms, enterococci, pseudomonads, fluorescent pseudomonads, yeasts and molds, and Staphylococcus aureus) were determined for minced meat, hamburgers, nuggets, white sausages, red sausages, escalope, and roll-ups. S. aureus isolates were tested for susceptibility to twenty antimicrobials of veterinary and human clinical significance (disc diffusion method, CLSI). Average microbial loads (log10 cfu/g) ranged from 2.63 ± 0.80 (enterococci) to 6.66 ± 1.09 (psychrotrophs). Average APCs (6.44 ± 1.16 log10 cfu/g) were regarded as acceptable according to EU microbiological criteria. The type of product had an influence (P < 0.05) on microbial loads, samples of escalope showing the highest counts for most microbial groups. Two-thirds (66.7%) of the samples tested harbored S. aureus. All the S. aureus isolates were multi-resistant (to between three and fifteen antibiotics). The greatest prevalence of resistance was shown for ampicillin, oxacillin, penicillin G, ceftazidime, and nalidixic acid. The results of this study show that poultry-based meat preparations present high microbial loads and are a major reservoir of antibiotic-resistant S. aureus strains. This highlights the need for correct handling of such foodstuffs with a view to reducing risks to consumers. © 2017 Poultry Science Association Inc.

Preliminary Efficacy Testing of the Disinfectant MicrobeCare XLP for Potential Use in Military Operational Environments.

PubMed

Madden, Jonathan F; Henrichs, Lori; Ervin, Mark D; Lospinoso, Joshua; Beachkofsky, Thomas M; Hardin, Carolyn A

2018-04-04

A safe, easy-to-use, permanently bonded antiseptic that does not require post-exposure bioload reduction but maintains effectiveness over time would have far-reaching implications across multiple industries. Health care is one such arena, particularly in austere military settings where resources are at a premium. MicrobeCare XLP (MicrobeCare, Buffalo Grove, IL, USA) is a commercially available spray-on agent that is advertised to covalently bond to surfaces and provide a long-lasting antimicrobial coating inhospitable to >99.99% of surface microorganisms. A pilot study was devised to gather baseline data regarding product efficacy and laboratory parameters before consideration of extended investigations and military utilization. The product manufacturer recommends bioload reductions before product application, following product application, and after each pathogenic exposure. To investigate the product's efficacy in circumstances more closely simulating a military operational setting in which post-pathogenic exposure bioload reduction would not be possible, this step was deliberately excluded from the test sequences. Using autoclaved surgical forceps, growth of Staphylococcus aureus and Acinetobacter baumannii was evaluated in a controlled manner under multiple conditions. Test variations included duration of submersion in the MicrobeCare XLP solution and air-drying and a second autoclave sterilization. Control and treated forceps were exposed to a bacterial suspension and air-dried before being submerged in sterile saline and vortex mixed. The saline solution was serially diluted and plated on tryptic soy agar (TSA) II plates. Plates were incubated for 24 h and bacterial colony-forming units (CFU)/mL were counted. Statistical significance was defined according to the American Society for Testing and Materials (ASTM) International passing criteria of 3 Log10 or 99.9% reduction of microorganisms. Additionally, p-values were calculated using two-tailed unpaired two-sample t-tests with unequal variance with a threshold of 0.05. In the S. aureus tests, none of the reduction calculations met the ASTM International passing criteria. In addition, the difference between the means of the colony counts in the MicrobeCare XLP-treated forceps and untreated control forceps was not statistically significant (p-value 0.109). Conversely, in the A. baumannii tests, each of the percent reduction calculations met the ASTM International passing criteria; the difference between the means of the colony counts in the treatment and control groups was statistically significant (p-value 0.008). In these independent tests, MicrobeCare XLP effectively prevented growth of A. baumannii but had unpredictable results suppressing S. aureus. These results may relate to inherent properties of the bacteria or autoclave exposure, although the manufacturer asserts that the coating withstands such degradation. Additional testing could be performed using a broader range of microorganisms and exposure to varying conditions including other sterilization methods.

Viability and biomass of Micrococcus luteus DE2008 at different salinity concentrations determined by specific fluorochromes and CLSM-image analysis.

PubMed

Puyen, Zully M; Villagrasa, Eduard; Maldonado, Juan; Esteve, Isabel; Solé, Antonio

2012-01-01

In previous studies, our group developed a method based on Confocal Laser Scanning Microscopy and Image Analysis (CLSM-IA) to analyze the diversity and biomass of cyanobacteria in microbial mats. However, this method cannot be applied to heterotrophic microorganisms, as these do not have autofluorescence. In this article, we present a method that combines CLSM-IA and Hoechst 33342 and SYTOX Green fluorochromes (FLU-CLSM-IA) to determine the viability and biomass of Micrococcus luteus DE2008, isolated from a saline microbial mat (Ebro Delta, Tarragona, Spain). The method has been applied to assess the effect of salinity on this microorganism. A reduction in viability and biomass (live cells) was observed as the salt concentration increases. The largest effect was at 100‰ NaCl with a cell death of 27.25% and a decrease in total and individual biomass of 39.75 and 0.009 mgC/cm(3), respectively, both with respect to optimal growth (10 ‰ NaCl). On the other hand, another important contribution of this article was that combining the FLU-CLSM-IA results with those achieved by plate counts enabled us to determine, for first time, the viability and the total biomass of the "dormant cells" (66.75% of viability and 40.59 mgC/cm(3) of total biomass at 100‰ NaCl). FLU-CLSM-IA is an efficient, fast, and reliable method for making a total count of cells at pixel level, including the dormant cells, to evaluate the viability and the biomass of a hetetrophic microorganism, M. luteus DE2008.

Transfer of Pathogens from Cantaloupe Rind to Preparation Surfaces and Edible Tissue as a Function of Cutting Method.

PubMed

Shearer, Adrienne E H; LeStrange, Kyle; Castañeda Saldaña, Rafael; Kniel, Kalmia E

2016-05-01

Whole and cut cantaloupes have been implicated as vehicles in foodborne illness outbreaks of norovirus, salmonellosis, and listeriosis. Preparation methods that minimize pathogen transfer from external surfaces to the edible tissue are needed. Two preparation methods were compared for the transfer of Listeria monocytogenes, Salmonella enterica serovar Typhimurium LT2, murine norovirus, and Tulane virus from inoculated cantaloupe rinds to edible tissue and preparation surfaces. For the first method, cantaloupes were cut into eighths, and edible tissue was separated from the rind and cubed with the same knife used to open the cantaloupes. For the second method, cantaloupes were scored with a knife around the circumference sufficient to allow manual separation of the cantaloupes into halves. Edible tissue was scooped with a spoon and did not contact the preparation surface touched by the rind. Bacteria and virus were recovered from the rinds, preparation surfaces, and edible tissue and enumerated by culture methods and reverse transcription, quantitative PCR, respectively. Standard plate counts were determined throughout refrigerated storage of cantaloupe tissue. Cut method 2 yielded approximately 1 log lower recovery of L. monocytogenes and Salmonella Typhimurium from edible tissue, depending on the medium in which the bacteria were inoculated. A slight reduction was observed in murine norovirus recovered from edible tissue by cut method 2. The Tulane virus was detected in approximately half of the sampled cantaloupe tissue and only at very low levels. Aerobic mesophilic colony counts were lower through day 6 of storage for buffered peptone water-inoculated cantaloupes prepared by cut method 2. No differences were observed in environmental contamination as a function of cutting method. Although small reductions in contamination of edible tissue were observed for cut method 2, the extent of microbial transfer underscores the importance of preventing contamination of whole cantaloupes.

An evaluation of a partial-walled laminar-flow operating room

PubMed Central

Whyte, W.; Shaw, B. H.; Freeman, M. A. R.

1974-01-01

This paper contains an assessment of the physical performance of a permanently installed down-flow laminar-flow operating room at the London Hospital. This system employs partial walls extending 0·76 m (2·5 ft.) from the ceiling, from which the air is allowed to issue freely downwards at an initial velocity of about 0·4 m./sec. (80 ft./min.). The usefulness of the partial wall, as compared with a free issuing system, was demonstrated and a comparison made with a fully walled system. It was shown that a fully walled system would be more efficient than a partial-walled system as there was a loss in air velocity of about 20-25% with the partial wall due to the nonconstrained flow of air. This loss would be reflected in an increase in airborne bacterial count and would mean that an increase of 20-25% in the air volume would be required to obtain the same conditions as with the full-walled system. Entrainment of contaminated air was demonstrated but it was concluded that this would be of little consequence in the centre of the clean area, i.e. at the wound site. Sterile instruments, etc., however, on the outside of the clean area, would be more liable to airborne contamination. Bacterial and dust airborne counts taken during total hip operations gave a very low average figure (0·3 bacteria/ft.3 or 10·5/m.3) from which we conclude that the system was about 30 times cleaner in terms of airborne bacteria than a well ventilated conventional operating-room. We concluded that although the partial-walled system was slightly less efficacious than a normal full-walled system, the freedom of movement and of communication for the operating team could in some circumstances outweigh this disadvantage. Sound levels were such that normal conversation was possible with little or no awareness of background noise. ImagesFig. 2Fig. 3Plate 2Plate 2Plate 3Plate 3Plate 1 PMID:4529595

Microbial Populations in Two Swamp Soils of South Carolina

Treesearch

David S. Priester; William R. Harms

1971-01-01

Microbial populations were counted in agar-plated samples of two swamp soils collected in summer and winter. Number of aerobic and anaerobic microorganisms differed significantly among the soils and between seasons. Alluvial soil from the river swamp was high in organic matter, N, K, Ca, and pH and averaged 88 million microorganisms per gram over the growing season....

7 CFR 58.938 - Physical requirements and microbiological limits for sweetened condensed milk.

Code of Federal Regulations, 2013 CFR

2013-01-01

... retain a definite form. (e) Microbiological limits. (1) Coliforms, less than 10 per gram; (2) yeasts, less than 5 per gram; (3) molds, less than 5 per gram; (4) total plate count, less than 1,000 per gram... amount of sediment retained on a lintine disc after a sample composed of 225 grams of product dissolved...

Development of microchannel plates in advanced wind-tunnel instrumentation

NASA Technical Reports Server (NTRS)

Feller, W. Bruce

1990-01-01

Microchannel plate (MCP) electron multiplier dynamic range has been increased 3 to 4 orders of magnitude at ambient temperatures, through enhanced input count rate capability and reduced background or 'dark' noise. The previous upper limit of roughly 10(exp 7) - 10(exp 8) cm(exp -2)s(exp -1) at ambient has been extended to levels approach 10(exp 10) cm(exp -2)s(exp -1) under continuous dc operation. The lower limit, previously set by an irreducible background component (approximately 0.6 cm(exp -2)s(exp -1)), has been lowered to the cosmic ray limit of .01 cm(exp -2)s(exp -1). The high end improvement was achieved by conductively cooling a very low resistance MCP by bonding it to a heat sink, while maintaining pulse-counting operation with multianode readouts. The low-end improvement was achieved by removing all radioisotopes from the MCP matrix glass. The detectors will benefit optical and mass spectrometry, flow visualization, plasma diagnostics, magnetometry, and other high signal flux applications. Very low MCP background noise will benefit X-ray and UV astronomy, medical imaging, trace isotope mass spectrometry, and other applications where the signal flux is often extremely low.

Radium-226 body burden in U miners by measurement of Rn in exhaled breath.

PubMed

Srivastava, G K; Raghavayya, M; Kotrappa, P; Somasundaram, S

1986-02-01

Uranium miners were made to inhale Rn-free medical O2 and exhale through a 5.2-1 A1 chamber before reporting to work. The chamber was sealed and isolated from the sampling circuit. An electrostatic plate collected the freshly formed Rn-decay products. The subsequent programmed alpha counting of the plate yielded a Rn concentration in the exhaled breath. Assuming that the exhaled breath represents a certain fraction of the Rn produced inside the body, the body burden of 226Ra was calculated. Standardisation of this procedure and the data collected on 310 miners are discussed. The procedure is simple and applicable for routine measurements. The miner needs to be in the laboratory for only 10 min. The system is also portable for field application. For routine use, the minimum detectable concentration is 3.87 Bq X m-3 which corresponds to a body burden of 0.26 kBq in a typical miner, if one assumes the Rn release fraction from the body as 84%. The system offers a more convenient and sensitive alternative to whole-body counting of workers for 226Ra.

Biogenic amine survey and organoleptic changes in fresh, stored, and temperature-abused bluefish (Pomatomus saltatrix).

PubMed

Gingerich, T M; Lorca, T; Flick, G J; Pierson, M D; McNair, H M

1999-09-01

Changes in histamine, putrescine, and cadaverine concentrations in bluefish filets (Pomatomus saltatrix) stored at 5, 10, and 15 degrees C were determined using high-performance liquid chromatography. An organoleptic assessment was conducted simultaneously with the biogenic amine analyses. The histamine levels found in fresh bluefish obtained from wholesale seafood distributors ranged between <1 ppm and 99 with an average of 39 ppm. Putrescine and cadaverine were not found in fresh bluefish. Fish fillets stored at each of the three temperatures developed histamine. The greatest accumulation of histamine was observed in fish stored at 15 degrees C, which developed histamine levels as high as 2,200 ppm. Putrescine levels increased at each temperature during storage. Cadaverine was present only in uninoculated bluefish stored at 15 degrees C. Histamine achieved higher levels in bluefish pieces inoculated with Morganella morganii, which demonstrates that bluefish support bacterial histamine formation. Histamine levels at each temperature exceeded the 50-ppm advisory level established by the Food and Drug Administration before 100% sensory rejection. Standard plate counts increased during storage of fish at all temperatures, but the correlation between histamine levels and standard plate count was not significant.

Nurses’ uniforms: How many bacteria do they carry after one shift?

PubMed Central

Sanon, Marie-Anne; Watkins, Sally

2013-01-01

This pilot study investigated the pathogens that nurses are potentially bringing into the public and their home when they wear work uniforms outside of the work environment. To achieve this, sterilized uniforms were distributed to 10 nurses at a local hospital in Washington State at the beginning of their shift. Worn uniforms were collected at the end of the shifts and sent to a laboratory for analysis. Four tests were conducted: 1) a heterotrophic growth plate count, 2) methicillin-resistant Staphylococcus aureus (MRSA) growth, 3) vancomycin-resistant Enterococci (VRE), and 4) identification of the heterotrophic plate counts. Each participant completed a questionnaire and a survey. The results showed that the average bacteria colony growth per square inch was 1,246 and 5,795 for day and night shift, respectively. After 48 h, MRSA positives were present on 4 of the day shift and 3 of the night shift uniforms. Additional bacteria identified include: Bacillus sp., Micrococcus luteus, Staphylococcus aureus, Staphylococcus epidermidis, and Micrococcus roseus. The significant presence of bacteria on the uniforms 48 h after the shift ended necessitates further study, discussions and policy consideration regarding wearing health care uniforms outside of the work environment. PMID:25285235

Nurses' uniforms: How many bacteria do they carry after one shift?

PubMed

Sanon, Marie-Anne; Watkins, Sally

2012-12-01

This pilot study investigated the pathogens that nurses are potentially bringing into the public and their home when they wear work uniforms outside of the work environment. To achieve this, sterilized uniforms were distributed to 10 nurses at a local hospital in Washington State at the beginning of their shift. Worn uniforms were collected at the end of the shifts and sent to a laboratory for analysis. Four tests were conducted: 1) a heterotrophic growth plate count, 2) methicillin-resistant Staphylococcus aureus (MRSA) growth, 3) vancomycin-resistant Enterococci (VRE), and 4) identification of the heterotrophic plate counts. Each participant completed a questionnaire and a survey. The results showed that the average bacteria colony growth per square inch was 1,246 and 5,795 for day and night shift, respectively. After 48 h, MRSA positives were present on 4 of the day shift and 3 of the night shift uniforms. Additional bacteria identified include: Bacillus sp., Micrococcus luteus, Staphylococcus aureus, Staphylococcus epidermidis, and Micrococcus roseus. The significant presence of bacteria on the uniforms 48 h after the shift ended necessitates further study, discussions and policy consideration regarding wearing health care uniforms outside of the work environment.

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

PubMed

Horn, Suranie; Pieters, Rialet; Bezuidenhout, Carlos

2016-12-01

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

Microbiological Quality Assessment of Game Meats at Retail in Japan.

PubMed

Asakura, Hiroshi; Kawase, Jun; Ikeda, Tetsuya; Honda, Mioko; Sasaki, Yoshimasa; Uema, Masashi; Kabeya, Hidenori; Sugiyama, Hiromu; Igimi, Shizunobu; Takai, Shinji

2017-12-01

In this study, we examined the prevalence of Shiga toxin-producing Escherichia coli and Salmonella spp. and the distribution of indicator bacteria in 248 samples of game meats (120 venison and 128 wild boar) retailed between November 2015 and March 2016 in Japan. No Salmonella spp. were detected in any of the samples, whereas Shiga toxin-producing Escherichia coli serotype OUT:H25 (stx 2d + , eae - ) was isolated from one deer meat sample, suggesting a possible source for human infection. Plate count assays indicated greater prevalence of coliforms and E. coli in wild boar meat than in venison, whereas their prevalence in processing facilities showed greater variation than in animal species. The 16S rRNA ion semiconductor sequencing analysis of 24 representative samples revealed that the abundances of Acinetobacter and Arthrobacter spp. significantly correlated with the prevalence of E. coli, and quantitative PCR analyses in combination with selective plate count assay verified these correlations. To our knowledge, this is the first report to characterize the diversity of microorganisms of game meats at retail in Japan, together with identification of dominant microbiota. Our data suggest the necessity of bottom-up hygienic assessment in areas of slaughtering and processing facilities to improve microbiological safety.

Efficacy of Sodium Hypochlorite and Acidified Sodium Chlorite in Preventing Browning and Microbial Growth on Fresh-Cut Produce

PubMed Central

Sun, Shih Hui; Kim, Su Jin; Kwak, Soo Jin; Yoon, Ki Sun

2012-01-01

The use of suitable sanitizers can increase the quality of fresh-cut produce and reduce the risk of foodborne illnesses. The objective of this study was to compare the washing effects of 100 mg/L sodium hypochlorite (SH) and 500 mg/L acidified sodium chlorite (ASC) on the prevention of enzymatic browning and the growth of microbial populations, including aerobic plate counts, E. coli, and coliforms, throughout storage at 4°C and 10°C. Fresh-cut zucchini, cucumbers, green bell peppers, and root vegetables such as potatoes, sweet potatoes, carrots, and radishes were used. Compared to SH washing, ASC washing significantly (p<0.05) reduced microbial contamination on the fresh-cut produce and prevented browning of fresh-cut potatoes and sweet potatoes during storage. More effective inhibition of aerobic plate counts and coliforms growth was observed on fresh-cut produce treated with ASC during storage at 10°C. Polyphenol oxidase (PPO) activity of fresh-cut potatoes and sweet potatoes was more effectively inhibited after washing with ASC. The use of 500 mg/L ASC can provide effective antimicrobial and anti-browning treatments of fresh-cut produce, including processed root vegetables. PMID:24471086

Efficacy of sodium hypochlorite and acidified sodium chlorite in preventing browning and microbial growth on fresh-cut produce.

PubMed

Sun, Shih Hui; Kim, Su Jin; Kwak, Soo Jin; Yoon, Ki Sun

2012-09-01

The use of suitable sanitizers can increase the quality of fresh-cut produce and reduce the risk of foodborne illnesses. The objective of this study was to compare the washing effects of 100 mg/L sodium hypochlorite (SH) and 500 mg/L acidified sodium chlorite (ASC) on the prevention of enzymatic browning and the growth of microbial populations, including aerobic plate counts, E. coli, and coliforms, throughout storage at 4°C and 10°C. Fresh-cut zucchini, cucumbers, green bell peppers, and root vegetables such as potatoes, sweet potatoes, carrots, and radishes were used. Compared to SH washing, ASC washing significantly (p<0.05) reduced microbial contamination on the fresh-cut produce and prevented browning of fresh-cut potatoes and sweet potatoes during storage. More effective inhibition of aerobic plate counts and coliforms growth was observed on fresh-cut produce treated with ASC during storage at 10°C. Polyphenol oxidase (PPO) activity of fresh-cut potatoes and sweet potatoes was more effectively inhibited after washing with ASC. The use of 500 mg/L ASC can provide effective antimicrobial and anti-browning treatments of fresh-cut produce, including processed root vegetables.

Effect of the implementation of HACCP on the microbiological quality of meals at a university restaurant.

PubMed

Cenci-Goga, B T; Ortenzi, R; Bartocci, E; Codega de Oliveira, A; Clementi, F; Vizzani, A

2005-01-01

A study was conducted to evaluate the microbiological quality, including total mesophilic counts and markers of bacteriological hygiene, as indicator of food safety of three categories of the most consumed meals in a university restaurant, before and after implementation of the HACCP system and personnel training. Cold gastronomy products, cooked warm-served products, and cooked cold-served products were tested for bacterial contamination. Throughout the experiment, 894 samples were examined for total counts of aerobic bacteria, counts of indicator organisms (coliform organisms and Escherichia coli) and pathogens (Staphylococcus aureus, Bacillus cereus, Salmonella spp., and Listeria monocytogenes). Implementation of the HACCP system, together with training in personnel hygiene, good manufacturing practices, and cleaning and sanitation procedures, resulted in lower aerobic plate counts and a lower incidence of S. aureus, coliform organisms, E. coli, and B. cereus, whereas Salmonella spp. and L. monocytogenes were not found in all samples studied. The microbial results of this study demonstrate that personnel training together with HACCP application contributed to improve the food safety of meals served in the restaurant studied.

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

PubMed

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

2015-10-01

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

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

NASA Astrophysics Data System (ADS)

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

2015-10-01

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

A System for Photon-Counting Spectrophotometry of Prompt Optical Emission from Gamma-Ray Bursts

NASA Astrophysics Data System (ADS)

Vestrand, W. T.; Albright, K.; Casperson, D.; Fenimore, E.; Ho, C.; Priedhorsky, W.; White, R.; Wren, J.

2003-04-01

With the launch of HETE-2 and the coming launch of the Swift satellite, there will be many new opportunities to study the physics of the prompt optical emission with robotic ground-based telescopes. Time-resolved spectrophotometry of the rapidly varying optical emission is likely to be a rich area for discovery. We describe a program to apply state-of-the-art photon-counting imaging technology to the study of prompt optical emission from gamma-ray bursts. The Remote Ultra-Low Light Imaging (RULLI) project at Los Alamos National Laboratory has developed an imaging sensor which employs stacked microchannel plates and a crossed delay line readout with 200 picosecond photon timing to measure the time of arrival and positions for individual optical photons. RULLI detectors, when coupled with a transmission grating having 300 grooves/mm, can make photon-counting spectroscopic observations with spectral resolution that is an order of magnitude greater and temporal resolution three orders of magnitude greater than the most capable photon-counting imaging detectors that have been used for optical astronomy.

Monitoring of biofilm-associated Legionella pneumophila on different substrata in model cooling tower system.

PubMed

Türetgen, Irfan; Cotuk, Aysin

2007-02-01

Cooling towers have the potential to develop infectious concentrations of Legionella pneumophila. Legionella counts increases where biofilm and warm water temperatures are present. In this study, biofilm associated L. pneumophila and heterotrophic bacteria were compared in terms of material dependence. Model cooling tower system was experimentally infected by L. pneumophila standard strain and monthly monitored. Different materials were tested for a period of 180 days. The lowest L. pneumophila and heterotrophic plate counts were measured on plastic polymers, whereas L. pneumophila and heterotrophic bacteria were accumulated rapidly on galvanized steel surfaces. It can be concluded that selection of plastic polymers, as a manufacturing material, are suitable for recirculating water systems.

High Throughput, High Yield Fabrication of High Quantum Efficiency Back-Illuminated Photon Counting, Far UV, UV, and Visible Detector Arrays

NASA Technical Reports Server (NTRS)

Nikzad, Shouleh; Hoenk, M. E.; Carver, A. G.; Jones, T. J.; Greer, F.; Hamden, E.; Goodsall, T.

2013-01-01

In this paper we discuss the high throughput end-to-end post fabrication processing of high performance delta-doped and superlattice-doped silicon imagers for UV, visible, and NIR applications. As an example, we present our results on far ultraviolet and ultraviolet quantum efficiency (QE) in a photon counting, detector array. We have improved the QE by nearly an order of magnitude over microchannel plates (MCPs) that are the state-of-the-art UV detectors for many NASA space missions as well as defense applications. These achievements are made possible by precision interface band engineering of Molecular Beam Epitaxy (MBE) and Atomic Layer Deposition (ALD).

Sub-Plate Overlap Code Documentation

NASA Technical Reports Server (NTRS)

Taff, L. G.; Bucciarelli, B.; Zarate, N.

1997-01-01

An expansion of the plate overlap method of astrometric data reduction to a single plate has been proposed and successfully tested. Each plate is (artificially) divided into sub-plates which can then be overlapped. This reduces the area of a 'plate' over which a plate model needs to accurately represent the relationship between measured coordinates and standard coordinates. Application is made to non-astrographic plates such as Schmidt plates and to wide-field astrographic plates. Indeed, the method is completely general and can be applied to any type of recording media.

A Study on the Saving Method of Plate Jigs in Hull Block Butt Welding

NASA Astrophysics Data System (ADS)

Ko, Dae-Eun

2017-11-01

A large amount of plate jigs is used for alignment of welding line and control of welding deformations in hull block assembly stage. Besides material cost, the huge working man-hours required for working process of plate jigs is one of the obstacles in productivity growth of shipyard. In this study, analysis method was proposed to simulate the welding deformations of block butt joint with plate jigs setting. Using the proposed analysis method, an example simulation was performed for actual panel block joint to investigate the saving method of plate jigs. Results show that it is possible to achieve two objectives of quality accuracy of the hull block and saving the plate jig usage at the same time by deploying the plate jigs at the right places. And the proposed analysis method can be used in establishing guidelines for the proper use of plate jigs in block assembly stage.

Elastostatic stress analysis of orthotropic rectangular center-cracked plates

NASA Technical Reports Server (NTRS)

Gyekenyesi, G. S.; Mendelson, A.

1972-01-01

A mapping-collocation method was developed for the elastostatic stress analysis of finite, anisotropic plates with centrally located traction-free cracks. The method essentially consists of mapping the crack into the unit circle and satisfying the crack boundary conditions exactly with the help of Muskhelishvili's function extension concept. The conditions on the outer boundary are satisfied approximately by applying the method of least-squares boundary collocation. A parametric study of finite-plate stress intensity factors, employing this mapping-collocation method, is presented. It shows the effects of varying material properties, orientation angle, and crack-length-to-plate-width and plate-height-to-plate-width ratios for rectangular orthotropic plates under constant tensile and shear loads.

Detection of License Plate using Sliding Window, Histogram of Oriented Gradient, and Support Vector Machines Method

NASA Astrophysics Data System (ADS)

Astawa, INGA; Gusti Ngurah Bagus Caturbawa, I.; Made Sajayasa, I.; Dwi Suta Atmaja, I. Made Ari

2018-01-01

The license plate recognition usually used as part of system such as parking system. License plate detection considered as the most important step in the license plate recognition system. We propose methods that can be used to detect the vehicle plate on mobile phone. In this paper, we used Sliding Window, Histogram of Oriented Gradient (HOG), and Support Vector Machines (SVM) method to license plate detection so it will increase the detection level even though the image is not in a good quality. The image proceed by Sliding Window method in order to find plate position. Feature extraction in every window movement had been done by HOG and SVM method. Good result had shown in this research, which is 96% of accuracy.

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.

PubMed

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.

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

NASA Astrophysics Data System (ADS)

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

2005-01-01

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

Large Area and High Efficiency Photon Counting Imaging Detectors with High Time and Spatial Resolution for Night Time Sensing and Astronomy

NASA Astrophysics Data System (ADS)

Siegmund, O.; Vallerga, J.; Tremsin, A.; McPhate, J.; Frisch, H.; Elam, J.; Mane, A.; Wagner, R.; Varner, G.

2012-09-01

The development of large area photon counting, imaging, timing detectors with high performance has significance for applications in astronomy (such as our sensor on the SAAO SALT 10m telescope), night time remote reconnaissance, airborne/space situational awareness, and high-speed adaptive optics. Sealed tube configurations for optical/IR sensing also have applications in detection of Cherenkov light (RICH), biological single-molecule fluorescence lifetime imaging microscopy and neutron imaging applications. In open faced configurations these devices are important for UV and particle detection in space astrophysics, mass spectroscopy and many time-of flight applications. Currently available devices are limited to sizes of about 5 cm and use either conventional microchannel plates, or dynode multipliers for amplification, coupled coarse pad array readouts. Extension of these schemes to devices as large as 20 cm with high spatial resolution presents significant problems and potentially considerable cost. A collaboration (Large Area Picosecond Photon Detector) of the U. Chicago, Argonne National Laboratory, U.C. Berkeley, U. Hawaii and a number of other institutions has developed novel technologies to realize 20 cm format detectors in open face or sealed tube configurations. One critical component of this development is novel microchannel plates employing borosilicate micro-capillary arrays. The microchannel plates are based on a novel concept where the substrate is constructed from a borosilicate micro-capillary array that is made to function as a microchannel plate by deposition of resistive and secondary emissive layers using atomic layer deposition. The process is relatively inexpensive compared with conventional microchannel plates and allows very large microchannel plates to be produced with pore sizes as small as 10 microns. These provide many performance characteristics typical of conventional microchannel plates, but have been made in sizes up to 20 cm, have low intrinsic background (<0.1 events/sq-cm/sec) and high stability with no observed gain degradation behavior over at least 5 Coul/sq-cm of charge extraction. Initial tests in a 20 cm detector with a cross strip electronic readout have achieved 4k x 4k pixel imaging with single photon sub-ns timing and MHz event rates. In concert with this effort we have made stable, uniform 20 cm bialkali photocathodes with >20% quantum efficiency on borosilicate windows compatible with a large sealed tube device. Other related efforts have also produced small sealed tubes with 30% quantum efficiency GaAs sealed tubes with high resolution imaging and timing that are immediately applicable to current applications, and opaque GaN UV photocathodes directly deposited onto these novel microchannel plates. We will discuss the details and implications of these novel microchannel plates with respect to the realization of novel detectors up to 20 cm format with reasonable cost and performance, robust construction, high flexibility of format and readout, reduction of fabrication effort, dramatically increased lifetime and stability, and their potential applications.

Comparison of dry sheet media and conventional agar media methods for enumerating yeasts and molds in food.

PubMed

Beuchat, L R; Mann, David A; Gurtler, Joshua B

2007-11-01

A study was done to compare Nissui Compact Dry Yeast and Mold plates (CDYM), 3M Petrifilm Yeast and Mold count plates (PYM), dichloran-rose bengal chloramphenicol (DRBC) agar, and dichloran 18% glycerol (DG18) agar for enumerating yeasts and molds naturally occurring in 97 foods (grains, legumes, raw fruits and vegetables, nuts, dairy products, meats, and miscellaneous processed foods and dry mixes). Correlation coefficients for plates incubated for 5 days were DG18 versus DRBC (0.93), PYM versus DRBC (0.81), CDYM versus DG18 (0.81), PYM versus DG18 (0.80), CDYM versus DRBC (0.79), and CDYM versus PYM (0.75). The number of yeasts and molds recovered from a group of foods (n = 32) analyzed on a weight basis (CFU per gram) was not significantly different (alpha = 0.05) when samples were plated on DRBC, DG18, PYM, or CDYM. However, the order of recovery from foods (n = 65) in a group analyzed on a unit or piece basis, or a composite of both groups (n = 97), was DRBC > DG18 = CDYM > PYM. Compared with PYM, CDYM recovered equivalent, significantly higher (alpha = 0.05) or significantly lower (alpha = 0.05) numbers of yeasts and molds in 51.5, 27.8, and 20.6%, respectively, of the 97 foods tested; respective values were 68.8, 15.6, and 15.6% in the small group (n = 32) and 43.1, 33.8, and 23.1% in the large group (n = 65) of foods. The two groups contained different types of foods, the latter consisting largely (73.8%) of raw fruits (n = 16) and vegetables (n = 32). Differences in efficacy of the four methods in recovering yeasts and molds from foods in the two groups are attributed in part to differences in genera and predominant mycoflora. While DG18 agar, CDYM, and PYM appear to be acceptable for enumerating yeasts and molds in the foods analyzed in this study, overall, DRBC agar recovered higher numbers from the 97 test foods, thereby supporting its recommended use as a general purpose medium for mycological analysis.

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

PubMed

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 (< 100 cfu.cm(-2)). S. aureus and Salmonella were not detected, but Listeria was detected in 23 % and E. coli in 1.3 % of the samples. Fifty percent (50 %) of the plants applied conventional cleaning methods for cleaning and sanitation and 50 % used the low-pressure foam (LPF) method. There was significant difference (P ≤ 0.05) between the mean TPC values of the conventional cleaning method (14 358.82) compared to that of LPF method (2 386.51) but no significant difference (P > 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-Sensitivity High-Speed X-ray Fluorescence Scanning Cadmium Telluride Detector for Deep-Portion Cancer Diagnosis Utilizing Tungsten-Kα-Excited Gadolinium Mapping

NASA Astrophysics Data System (ADS)

Yanbe, Yutaka; Sato, Eiichi; Chiba, Hiraku; Maeda, Tomoko; Matsushita, Ryo; Oda, Yasuyuki; Hagiwara, Osahiko; Matsukiyo, Hiroshi; Osawa, Akihiro; Enomoto, Toshiyuki; Watanabe, Manabu; Kusachi, Shinya; Sato, Shigehiro; Ogawa, Akira

2013-09-01

X-ray fluorescence (XRF) analysis is useful for mapping various atoms in objects. Bremsstrahlung X-rays with energies beyond tantalum (Ta) K-edge energy 67.4 keV are absorbed effectively using a 100-µm-thick Ta filter, and the filtered X-rays including tungsten (W) Kα rays are absorbed by gadolinium (Gd) atoms in objects. The Gd XRF is then produced from Gd atoms in the objects and is counted by a cadmium telluride (CdTe) detector. Gd Kα photons with a maximum count rate of 1 kilo counts per second are dispersed using a multichannel analyzer, and the number of photons is counted by a counter card. The distance between the CdTe detector and the object is minimized to 40 mm to increase the count rate. The object is scanned using an x-y stage with a velocity of 5.0 mm/s, and Gd mapping are shown on a computer monitor. The scan steps of the x- and y-axes were both 2.5 mm, and the photon-counting time per mapping point was 0.5 s. We obtained Gd XRF images at high contrast, and Gd Kα photons were easily detected from cancerous regions in a nude mouse placed behind a 20-mm-thick poly(methyl methacrylate) plate.

Comparison between two time-resolved approaches for prostate cancer diagnosis: high rate imager vs. photon counting system

NASA Astrophysics Data System (ADS)

Boutet, J.; Debourdeau, M.; Laidevant, A.; Hervé, L.; Dinten, J.-M.

2010-02-01

Finding a way to combine ultrasound and fluorescence optical imaging on an endorectal probe may improve early detection of prostate cancer. A trans-rectal probe adapted to fluorescence diffuse optical tomography measurements was developed by our team. This probe is based on a pulsed NIR laser source, an optical fiber network and a time-resolved detection system. A reconstruction algorithm was used to help locate and quantify fluorescent prostate tumors. In this study, two different kinds of time-resolved detectors are compared: High Rate Imaging system (HRI) and a photon counting system. The HRI is based on an intensified multichannel plate and a CCD Camera. The temporal resolution is obtained through a gating of the HRI. Despite a low temporal resolution (300ps), this system allows a simultaneous acquisition of the signal from a large number of detection fibers. In the photon counting setup, 4 photomultipliers are connected to a Time Correlated Single Photon Counting (TCSPC) board, providing a better temporal resolution (0.1 ps) at the expense of a limited number of detection fibers (4). At last, we show that the limited number of detection fibers of the photon counting setup is enough for a good localization and dramatically improves the overall acquisition time. The photon counting approach is then validated through the localization of fluorescent inclusions in a prostate-mimicking phantom.

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

PubMed

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.

Models of convection-driven tectonic plates - A comparison of methods and results

NASA Technical Reports Server (NTRS)

King, Scott D.; Gable, Carl W.; Weinstein, Stuart A.

1992-01-01

Recent numerical studies of convection in the earth's mantle have included various features of plate tectonics. This paper describes three methods of modeling plates: through material properties, through force balance, and through a thin power-law sheet approximation. The results obtained are compared using each method on a series of simple calculations. From these results, scaling relations between the different parameterizations are developed. While each method produces different degrees of deformation within the surface plate, the surface heat flux and average plate velocity agree to within a few percent. The main results are not dependent upon the plate modeling method and herefore are representative of the physical system modeled.

Effects of Jet Fuel Spills on the Microbial Community of Soil †

PubMed Central

Song, Hong-Gyu; Bartha, Richard

1990-01-01

Hydrocarbon residues, microbial numbers, and microbial activity were measured and correlated in loam soil contaminated by jet fuel spills resulting in 50 and 135 mg of hydrocarbon g of soil−1. Contaminated soil was incubated at 27°C either as well-aerated surface soil or as poorly aerated subsurface soil. In the former case, the effects of bioremediation treatment on residues, microbial numbers, and microbial activity were also assessed. Hydrocarbon residues were measured by quantitative gas chromatography. Enumerations included direct counts of metabolically active bacteria, measurement of mycelial length, plate counts of aerobic heterotrophs, and most probable numbers of hydrocarbon degraders. Activity was assessed by fluorescein diacetate (FDA) hydrolysis. Jet fuel disappeared much more rapidly from surface soil than it did from subsurface soil. In surface soil, microbial numbers and mycelial length were increased by 2 to 2.5 orders of magnitude as a result of jet fuel contamination alone and by 3 to 4 orders of magnitude as a result of the combination of jet fuel contamination and bioremediation. FDA hydrolysis was stimulated by jet fuel and bioremediation, but was inhibited by jet fuel alone. The latter was traced to an inhibition of the FDA assay by jet fuel biodegradation products. In subsurface soil, oxygen limitation strongly attenuated microbial responses to jet fuel. An increase in the most probable numbers of hydrocarbon degraders was accompanied by a decline in other aerobic heterotrophs, so that total plate counts changed little. The correlations between hydrocarbon residues, microbial numbers, and microbial activity help in elucidating microbial contributions to jet fuel elimination from soil. PMID:16348138

Computer vision-based carbohydrate estimation for type 1 patients with diabetes using smartphones.

PubMed

Anthimopoulos, Marios; Dehais, Joachim; Shevchik, Sergey; Ransford, Botwey H; Duke, David; Diem, Peter; Mougiakakou, Stavroula

2015-05-01

Individuals with type 1 diabetes (T1D) have to count the carbohydrates (CHOs) of their meal to estimate the prandial insulin dose needed to compensate for the meal's effect on blood glucose levels. CHO counting is very challenging but also crucial, since an error of 20 grams can substantially impair postprandial control. The GoCARB system is a smartphone application designed to support T1D patients with CHO counting of nonpacked foods. In a typical scenario, the user places a reference card next to the dish and acquires 2 images with his/her smartphone. From these images, the plate is detected and the different food items on the plate are automatically segmented and recognized, while their 3D shape is reconstructed. Finally, the food volumes are calculated and the CHO content is estimated by combining the previous results and using the USDA nutritional database. To evaluate the proposed system, a set of 24 multi-food dishes was used. For each dish, 3 pairs of images were taken and for each pair, the system was applied 4 times. The mean absolute percentage error in CHO estimation was 10 ± 12%, which led to a mean absolute error of 6 ± 8 CHO grams for normal-sized dishes. The laboratory experiments demonstrated the feasibility of the GoCARB prototype system since the error was below the initial goal of 20 grams. However, further improvements and evaluation are needed prior launching a system able to meet the inter- and intracultural eating habits. © 2015 Diabetes Technology Society.

Aseptic laboratory techniques: plating methods.

PubMed

Sanders, Erin R

2012-05-11

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

Evaluation of microbial survival post-incidence on fresh Mozzarella cheese.

PubMed

Ganesan, Balasubramanian; Irish, David A; Brothersen, Carl; McMahon, Donald J

2012-12-01

Commercial fresh Mozzarella cheese is made by direct acidification and is stored dry or in water without salt addition. The cheese has a shelf life of 6 wk, but usually develops an off-flavor and loses textural integrity by 4 wk, potentially due to the lack of salt and high moisture that allow the outgrowth of undesirable bacteria. To understand how microbial incidence affects cheese quality and how incident pathogen-related bacteria are limited by salt level during refrigerated storage, we made fresh Mozzarella cheese with high (2%) and low (0.5%) salt. The high-salt cheese was packaged and stored dry. The low-salt cheese was packaged and stored either dry or in 0.5% salt brine. One portion of cheeses was evaluated for surviving incident microbes by aerobic plate counts, coliform counts, and psychrophilic bacterial counts, of which coliforms and psychrophiles were not detected over 9 wk. Aerobic plate counts remained at 100 to 300 cfu/g up to 2 wk but increased by 1,000- to 10,000-fold between 4 and 6 wk at all salt levels and storage conditions. Other portions of cheeses were inoculated with either Escherichia coli or Enterococcus faecalis, both of which increased by 100-fold over 90 d of storage. Interestingly, E. coli added to the cheese brine first grew in the brine by 100-fold before attaching to the cheese, whereas Ent. faecalis attached to the cheese within 24h and grew only on the cheese. We conclude that incident bacteria, even from similar environments, may attach to cheese curd and survive differently in fresh Mozzarella cheese than in brine. Overall, 2% salt was insufficient to control bacterial growth, and slow-growing, cold- and salt-tolerant bacteria may survive and spoil fresh Mozzarella cheese. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Analysis of bacterial community shifts in the gastrointestinal tract of pigs fed diets supplemented with β-glucan from Laminaria digitata, Laminaria hyperborea and Saccharomyces cerevisiae.

PubMed

Murphy, P; Dal Bello, F; O'Doherty, J; Arendt, E K; Sweeney, T; Coffey, A

2013-07-01

This study was designed to evaluate the effects of algal and yeast β-glucans on the porcine gastrointestinal microbiota, specifically the community of Lactobacillus, Bifidobacterium and coliforms. A total of 48 pigs were fed four diets over a 28-day period to determine the effect that each had on these communities. The control diet consisted of wheat and soya bean meal. The remaining three diets contained wheat and soya bean meal supplemented with β-glucan at 250 g/tonne from Laminaria digitata, Laminaria hyperborea or Saccharomyces cerevisiae. Faecal samples were collected from animals before feeding each diet and after the feeding period. The animals were slaughtered the following day and samples were collected from the stomach, ileum, caecum, proximal colon and distal colon. Alterations in Lactobacillus in the gastrointestinal tract (GIT) were analysed using denaturing gradient gel electrophoresis (DGGE) profiles generated by group-specific 16S rRNA gene PCR amplicons. Plate count analysis was also performed to quantify total coliforms. DGGE profiles indicated that all β-glucan diets provoked the emergence of a richer community of Lactobacillus. The richest community of lactobacilli emerged after feeding L. digitata (LD β-glucan). Plate count analysis revealed that the L. hyperborea (LH β-glucan) diet had a statistically significant effect on the coliform counts in the proximal colon in comparison with the control diet. β-glucan from L. digitata and S. cerevisiae also generally reduced coliforms but to a lesser extent. Nevertheless, the β-glucan diets did not significantly reduce levels of Lactobacillus or Bifidobacterium. DGGE analysis of GIT samples indicated that the three β-glucan diets generally promoted the establishment of a more varied range of Lactobacillus species in the caecum, proximal and distal colon. The LH β-glucan had the most profound reducing effect on coliform counts when compared with the control diet and diets supplemented with L. digitata and S. cerevisiae β-glucans.

Colostrum and milk pasteurization improve health status and decrease mortality in neonatal calves receiving appropriate colostrum ingestion.

PubMed

Armengol, Ramon; Fraile, Lorenzo

2016-06-01

The objective of the study was to evaluate if on-farm heat treatment of colostrum and bulk tank milk can improve calf health status and morbidity and mortality rates during the first 21d of life in neonatal Holstein calves receiving appropriate colostrum ingestion. A total of 587 calves were randomly assigned to 2 groups of males and females over 18mo. The nonpasteurized group (n=287, 143 males and 144 females) was fed frozen (-20°C) colostrum (6-8L during the first 12h of life) that was previously reheated up to 40°C. They were also fed refrigerated (4°C) raw milk from the bulk tank that was also reheated up to 40°C (1.8L every 12h). The pasteurized group (n=300, 150 males and 150 females) was also fed colostrum and milk, but both were pasteurized before freezing. Blood samples were drawn from all calves to obtain serum at 2 to 5d of life. Serum total protein (g/dL) was determined using a commercially available refractometer. Colostrum and milk underwent routine bacteriological analysis to determine total plate counts (cfu/mL) and total coliform counts (cfu/mL). All the calves underwent clinical examination every 24h during the first 21d of life. Every day, calves were clinically diagnosed either as being healthy or suffering from respiratory disease, neonatal calf diarrhea, or suffering other diseases. On-farm heat treatment for colostrum and milk reduced total plate counts and total coliform counts between 1 and 2 log10. Pasteurization of colostrum and milk significantly decreased the morbidity and mortality (5.2 and 2.8%) in comparison with calves receiving nonpasteurized colostrum and milk (15.0 and 6.5%), respectively, during the first 21d of life, even in animals receiving appropriate colostrum ingestion. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Preliminary Evaluation of the Control of Microbial Fouling by Laboratory and Pilot-Scale Air-Stripping Columns

DTIC Science & Technology

1985-03-01

used to remove trichloroethylene (TCE) from contaminated well water. 7 MATERIALS Chemicals: Trichloroethylene (Aldrich chemical, Milwaukee, WI), sodium ...Cleveland, OH), sodium hydrcxide (J.T. Baker, Phillipsburg, NJ), potassium dichloroisocyanurate (Dorex Inc., Frankfort, IL), potassium iodide starch...NJ). Media and Reagents: Plate count agar (Difco Laboratories, Detroit, MI), lauryl tryptose broth (Difco Laboratories, Detroit, MI), motility medium

LENGTH-HETEROGENEITY POLYMERASE CHAIN REACTION (LH-PCR) AS AN INDICATOR OF STREAM SANITARY AND ECOLOGICAL CONDITION: OPTIMAL SAMPLE SIZE AND HOLDING CONDITIONS

EPA Science Inventory

The use of coliform plate count data to assess stream sanitary and ecological condition is limited by the need to store samples at 4oC and analyze them within a 24-hour period. We are testing LH-PCR as an alternative tool to assess the bacterial load of streams, offering a cost ...

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

NASA Astrophysics Data System (ADS)

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.

[Analysis of the microbiological quality and potential presence of Listeria monocytogenes in custard apple (Annona muricata), mango (Mangifera indica) and passion fruit (Passiflora edulis) pulps from Costa Rica].

PubMed

von Breymann, Juliana; Chaves, Carolina; Arias, María Laura

2013-03-01

The objective of this work was to determine some of the indicators associated to shelf life, hygiene, process and storage conditions for some of custard apple, mango and passion fruit pulps distributed by the main supermarket chains of the Metropolitan Area of San José, Costa Rica, as well as to examine the potential presence of Listeria monocytogenes in them. Sixty fruit pulp samples were analyzed. Tests included pH determination, total aerobic plate count, yeasts and mold count, lactic bacteria count, total and fecal most probable number and the presence/absence of Listeria monocytogenes in 25 g of the product. Fruit pulp's pH ranged between 3,1 and 3,9, and the microbiological counts obtained were relatively low except for one industry. None of the samples analyzed presented total or fecal coliforms. The presence of Listeria monocytogenes was confirmed in three samples, all of them coming from industry C. Low microbiological counts obtained may be due to the addition of preserving substances and to the pasteurization of some of the products; lack of these two elements may allow the presence of dangerous bacteria such as Listeria monocytogenes.

Colonisation of soilless growing media for tomato by Trichoderma harzianum.

PubMed

De Schutter, B; Aerts, R; Rombouts, L

2001-01-01

An experiment was conducted to evaluate the distribution of T. harzianum in soilless media used in greenhouse growing systems for tomatoes. Growing media based on rockwool and based on coconut fibre were included. The fungus was applied to the roots of the plant by means of a conidial suspension. The upper plant parts were removed from the coconut fibre an the rockwool slab after 10 and 15 weeks respectively. Both the coconut fibre and the rockwool slab were divided into fragments with a width of 10 cm. The coconut fibre medium fragments were again divided in an upper and a lower part. For every fragment the CFU/g was determined by two different methods. In the first method a known volume of growing medium was brought into suspension using a blender and diluted. Appropriate dilutions were plated on a selective medium. In the second method a known volume of growing medium was spread directly upon a selective medium in petri dishes. All dishes were incubated at room temperature and colonies were counted. Results showed that in the case of the coconut fibre growing medium T. harzianum was isolated in every fragment of the substrate. Highest densities were measured at the site of inoculation. In the lower part of the fragments less CFU/g were counted than in the upper part. In the case of the rockwool growing medium, there were several fragments in which no T. harzianum was isolated. The dilution technique demonstrated to be most useful in cases of high density. The direct spreading-method is best applied when low densities are expected. These observations demonstrate that the growing medium based upon coconut fibre is more appropriate for colonisation by T. harzianum. However, densities are higher at the site of inoculation in both tested growing media.

Possible overestimation of surface disinfection efficiency by assessment methods based on liquid sampling procedures as demonstrated by in situ quantification of spore viability.

PubMed

Grand, I; Bellon-Fontaine, M-N; Herry, J-M; Hilaire, D; Moriconi, F-X; Naïtali, M

2011-09-01

The standard test methods used to assess the efficiency of a disinfectant applied to surfaces are often based on counting the microbial survivors sampled in a liquid, but total cell removal from surfaces is seldom achieved. One might therefore wonder whether evaluations of microbial survivors in liquid-sampled cells are representative of the levels of survivors in whole populations. The present study was thus designed to determine the "damaged/undamaged" status induced by a peracetic acid disinfection for Bacillus atrophaeus spores deposited on glass coupons directly on this substrate and to compare it to the status of spores collected in liquid by a sampling procedure. The method utilized to assess the viability of both surface-associated and liquid-sampled spores included fluorescence labeling with a combination of Syto 61 and Chemchrome V6 dyes and quantifications by analyzing the images acquired by confocal laser scanning microscopy. The principal result of the study was that the viability of spores sampled in the liquid was found to be poorer than that of surface-associated spores. For example, after 2 min of peracetic acid disinfection, less than 17% ± 5% of viable cells were detected among liquid-sampled cells compared to 79% ± 5% or 47% ± 4%, respectively, when the viability was evaluated on the surface after or without the sampling procedure. Moreover, assessments of the survivors collected in the liquid phase, evaluated using the microscopic method and standard plate counts, were well correlated. Evaluations based on the determination of survivors among the liquid-sampled cells can thus overestimate the efficiency of surface disinfection procedures.

Possible Overestimation of Surface Disinfection Efficiency by Assessment Methods Based on Liquid Sampling Procedures as Demonstrated by In Situ Quantification of Spore Viability ▿

PubMed Central

Grand, I.; Bellon-Fontaine, M.-N.; Herry, J.-M.; Hilaire, D.; Moriconi, F.-X.; Naïtali, M.

2011-01-01

The standard test methods used to assess the efficiency of a disinfectant applied to surfaces are often based on counting the microbial survivors sampled in a liquid, but total cell removal from surfaces is seldom achieved. One might therefore wonder whether evaluations of microbial survivors in liquid-sampled cells are representative of the levels of survivors in whole populations. The present study was thus designed to determine the “damaged/undamaged” status induced by a peracetic acid disinfection for Bacillus atrophaeus spores deposited on glass coupons directly on this substrate and to compare it to the status of spores collected in liquid by a sampling procedure. The method utilized to assess the viability of both surface-associated and liquid-sampled spores included fluorescence labeling with a combination of Syto 61 and Chemchrome V6 dyes and quantifications by analyzing the images acquired by confocal laser scanning microscopy. The principal result of the study was that the viability of spores sampled in the liquid was found to be poorer than that of surface-associated spores. For example, after 2 min of peracetic acid disinfection, less than 17% ± 5% of viable cells were detected among liquid-sampled cells compared to 79% ± 5% or 47% ± 4%, respectively, when the viability was evaluated on the surface after or without the sampling procedure. Moreover, assessments of the survivors collected in the liquid phase, evaluated using the microscopic method and standard plate counts, were well correlated. Evaluations based on the determination of survivors among the liquid-sampled cells can thus overestimate the efficiency of surface disinfection procedures. PMID:21742922

Systems and Methods for the Electrodeposition of a Nickel-cobalt Alloy

NASA Technical Reports Server (NTRS)

Ogozalek, Nance Jo (Inventor); Wistrand, Richard E. (Inventor)

2013-01-01

Systems and methods for electrodepositing a nickel-cobalt alloy using a rotating cylinder electrode assembly with a plating surface and an electrical contact. The assembly is placed within a plating bath and rotated while running a plating cycle. Nickel-cobalt alloy deposition is selectively controlled by controlling current density distribution and/or cobalt content in the plating bath while running the plating cycle to deposit an alloy of a desired yield strength onto the plating surface in a single plating cycle. In various embodiments, the rotating cylinder may be used as an insitu monitoring method to assist in obtaining the properties desired.

Comment on: 'A Poisson resampling method for simulating reduced counts in nuclear medicine images'.

PubMed

de Nijs, Robin

2015-07-21

In order to be able to calculate half-count images from already acquired data, White and Lawson published their method based on Poisson resampling. They verified their method experimentally by measurements with a Co-57 flood source. In this comment their results are reproduced and confirmed by a direct numerical simulation in Matlab. Not only Poisson resampling, but also two direct redrawing methods were investigated. Redrawing methods were based on a Poisson and a Gaussian distribution. Mean, standard deviation, skewness and excess kurtosis half-count/full-count ratios were determined for all methods, and compared to the theoretical values for a Poisson distribution. Statistical parameters showed the same behavior as in the original note and showed the superiority of the Poisson resampling method. Rounding off before saving of the half count image had a severe impact on counting statistics for counts below 100. Only Poisson resampling was not affected by this, while Gaussian redrawing was less affected by it than Poisson redrawing. Poisson resampling is the method of choice, when simulating half-count (or less) images from full-count images. It simulates correctly the statistical properties, also in the case of rounding off of the images.

Advanced technology development multi-color holography

NASA Technical Reports Server (NTRS)

Vikram, Chandra S.

1993-01-01

This is the final report of the Multi-color Holography project. The comprehensive study considers some strategic aspects of multi-color holography. First, various methods of available techniques for accurate fringe counting are reviewed. These are heterodyne interferometry, quasi-heterodyne interferometry, and phase-shifting interferometry. Phase-shifting interferometry was found to be the most suitable for multi-color holography. Details of experimentation with a sugar solution are also reported where better than 1/200 of a fringe order measurement capability was established. Rotating plate glass phase shifter was used for the experimentation. The report then describes the possible role of using more than two wavelengths with special reference-to-object beam intensity ratio needs in multicolor holography. Some specific two- and three-color cases are also described in detail. Then some new analysis methods of the reconstructed wavefront are considered. These are deflectometry, speckle metrology, confocal optical signal processing, and phase shifting technique related applications. Finally, design aspects of an experimental breadboard are presented.

Puesta en marcha de un microdensitómetro automático basado en CCD

NASA Astrophysics Data System (ADS)

Calderón, J. H.; Bustos Fierro, I. H.

We present the commisioning of a CCD-based microdensitometer intended to perform astrometric measurements of photographic plates. The work done consisted in the installation of a CCD camera, the modification of the motion system, the construction of a new illumination device, the adaptation of the electronics, and the development of software. The instrument is intended to be used for the astrometric measurement mainly of plates of the Astrographic Catalog and Carte du Ciel collections from Córdoba Observatory. In this phase of the project we counted with the collaboration of the Instituto Provincial de Enseñanza Media No 59, 25 de Mayo, Cruz Alta (Province of Córdoba). The origin and importance of such collaboration is commented.

Aftershocks to Philippine quake found within nearby megathrust fault

NASA Astrophysics Data System (ADS)

Schultz, Colin

2013-02-01

On 31 August 2012 a magnitude 7.6 earthquake ruptured deep beneath the sea floor of the Philippine Trench, a powerful intraplate earthquake centered seaward of the plate boundary. In the wake of the main shock, sensors detected a flurry of aftershocks, counting 110 in total. Drawing on seismic wave observations and rupture mechanisms calculated for the aftershocks, Ye et al. found that many were located near the epicenter of the main intraplate quake but at shallower depth; all involved normal faulting. Some shallow thrusting aftershocks were located farther to the west, centered within the potentially dangerous megathrust fault formed by the subduction of the Philippine Sea plate beneath the Philippine microplate, the piece of crust housing the Philippine Islands.

Aerial ultrasound source with a circular vibrating plate attached to a rigid circumferential wall

NASA Astrophysics Data System (ADS)

Kuratomi, Ryo; Asami, Takuya; Miura, Hikaru

2018-07-01

We fabricate a transverse vibrating plate attached to a rigid wall integrated at the circumference of a circular vibrating plate that allows a strong sound wave field to be formed in the area encoded by the vibrating plate and rigid wall by installing a wall such as a reflective plate on the rigid wall. The design method for the circular vibrating plate attached to a rigid circumferential wall is investigated. A method of forming a strong standing wave field in an enclosed area constructed with a vibrating plate, cylindrical reflective plate, and parallel reflective plate is developed.

Cryopreservation on a cryo-plate of Arundina graminifolia protocorms, dehydrated with silica gel and drying beads.

PubMed

Cordova, L B; Thammasiri, K

2016-01-01

There are various methods for the cryopreservation of plant material, with each biological specimen potentially requiring protocol optimization to maximize success. The aim of this study is to compare droplet-vitrification, encapsulation-dehydration, and the cryo-plate method for cryopreservation of protocorms of the orchid Arundina graminifolia, using silica gel and drying beads as the desiccation materials. The cryo-plate method included preculture of protocorms, developed from seeds, placed on aluminium cryo-plates and embedded in alginate gel. Cryo-plates were surface dried using sterile filter paper, placed in Petri dishes containing 50 g silica gel or 30 g drying beads in a laminar air-flow cabinet. Specimens on cryo-plates were dehydrated to 25 % moisture content, placed into 2 mL cryotubes and plunged directly into liquid nitrogen for 1 d. For cryopreservation, the cryo-plate method, involving dehydration with 30 g drying beads gave the highest regrowth (77 %), followed by the encapsulation-dehydration method with 30 g drying beads (64 % regrowth) and the droplet-vitrification method, following exposure to PVS2 solution for 20 min (33 % regrowth). Regrowth of cryopreserved protocorms using the cryo-plate method was rapid with the highest survival and regrowth.

Ultra-compact imaging plate scanner module using a MEMS mirror and specially designed MPPC

NASA Astrophysics Data System (ADS)

Miyamoto, Yuichi; Sasaki, Kensuke; Takasaka, Masaomi; Fujimoto, Masatoshi; Yamamoto, Koei

2017-02-01

Computed radiography (CR), which is one of the most useful methods for dental imaging and nondestructive testing, uses a phosphor imaging plate (IP) because it is flexible, reusable, and inexpensive. Conventional IP scanners utilize a galvanometer or a polygon mirror as a scanning device and a photomultiplier as an optical sensor. Microelectromechanical systems (MEMS) technology currently provides silicon-based devices and has the potential to replace such discrete devices and sensors. Using these devices, we constructed an ultra-compact IP scanner. Our extremely compact plate scanner utilizes a module that is composed of a one-dimensional MEMS mirror and a long multi-pixel photon counter (MPPC) that is combined with a specially designed wavelength filter and a rod lens. The MEMS mirror, which is a non-resonant electromagnetic type, is 2.6 mm in diameter with a recommended optical scanning angle up to +/-15°. The CR's wide dynamic range is maintained using a newly developed MPPC. The MPPC is a sort of silicon photomultiplier and is a high-sensitivity photon-counting device. To achieve such a wide dynamic range, we developed a long MPPC that has over 10,000 pixels. For size reduction and high optical efficiency, we set the MPPC close to an IP across the rod lens. To prevent the MPPC from detecting excitation light, which is much more intense than photo-stimulated light, we produced a sharp-cut wavelength filter that has a wide angle (+/-60°) of tolerance. We evaluated our constructed scanner module through gray chart and resolution chart images.

Antibacterial properties of 2% lidocaine and reduced rate of endophthalmitis after intravitreal injection.

PubMed

Tustin, Aaron; Kim, Stephen J; Chomsky, Amy; Hubbard, G Baker; Sheng, Jinsong

2014-05-01

To determine whether the application of subconjunctival 2% lidocaine/0.1% methylparaben for anesthesia may reduce rates of endophthalmitis after intravitreal (IVT) injection. We performed in vitro experiments to determine the antibacterial properties of 2% lidocaine/0.1% methylparaben (lidocaine) against causative organisms of endophthalmitis. Isolates of Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus viridans from patients with endophthalmitis were incubated with or without lidocaine. Aliquots (100 µL) were plated on Mueller-Hinton (S. aureus and S. epidermidis) or blood agar plates (S. viridans) at 0, 10, 30, 120, and 240 minutes, and colonies were counted after 24 hours. A retrospective review of 15,042 IVT injections was performed from January 2004 to February 2011 to determine the rate of endophthalmitis with or without application of subconjunctival lidocaine for anesthesia. Lidocaine demonstrated rapid bactericidal effects against all 3 organisms. After 10 minutes of exposure, there was approximately a 90% (P < 0.01), 95% (P < 0.001), and 92% (P < 0.001) reduction in colony forming units when compared with time 0 for S. aureus, S. epidermidis, and S. viridans, respectively. Complete elimination of colony forming units occurred at subsequent time points for each organism in contrast to logarithmic increase for control plates. There were a total of 0 cases of endophthalmitis of 6,853 IVT injections performed with subconjunctival lidocaine and 8 cases of endophthalmitis of 8,189 (0.1%) IVT injections performed with other methods of anesthesia (P = 0.03). Application of subconjunctival 2% lidocaine/0.1% methylparaben for anesthesia may reduce the incidence of endophthalmitis after IVT injection.

Current Perspectives on Viable but Non-culturable State in Foodborne Pathogens

PubMed Central

Zhao, Xihong; Zhong, Junliang; Wei, Caijiao; Lin, Chii-Wann; Ding, Tian

2017-01-01

The viable but non-culturable (VBNC) state, a unique state in which a number of bacteria respond to adverse circumstances, was first discovered in 1982. Unfortunately, it has been reported that many foodborne pathogens can be induced to enter the VBNC state by the limiting environmental conditions during food processing and preservation, such as extreme temperatures, drying, irradiation, pulsed electric field, and high pressure stress, as well as the addition of preservatives and disinfectants. After entering the VBNC state, foodborne pathogens will introduce a serious crisis to food safety and public health because they cannot be detected using conventional plate counting techniques. This review provides an overview of the various features of the VBNC state, including the biological characteristics, induction and resuscitation factors, formation and resuscitation mechanisms, detection methods, and relationship to food safety. PMID:28421064

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

PubMed Central

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

1970-01-01

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

High altitude hypoxia as a factor that promotes tibial growth plate development in broiler chickens

PubMed Central

Huang, Shucheng; Zhang, Lihong; Rehman, Mujeeb Ur; Iqbal, Muhammad Kashif; Lan, Yanfang; Mehmood, Khalid; Zhang, Hui; Qiu, Gang; Nabi, Fazul; Yao, Wangyuan; Wang, Meng; Li, Jiakui

2017-01-01

Tibial dyschondroplasia (TD) is one of the most common problems in the poultry industry and leads to lameness by affecting the proximal growth plate of the tibia. However, due to the unique environmental and geographical conditions of Tibet, no case of TD has been reported in Tibetan chickens (TBCs). The present study was designed to investigate the effect of high altitude hypoxia on blood parameters and tibial growth plate development in chickens using the complete blood count, morphology, and histological examination. The results of this study showed an undesirable impact on the overall performance, body weight, and mortality of Arbor Acres chickens (AACs) exposed to a high altitude hypoxic environment. However, AACs raised under hypoxic conditions showed an elevated number of red blood cells (RBCs) and an increase in hemoglobin and hematocrit values on day 14 compared to the hypobaric normoxia group. Notably, the morphology and histology analyses showed that the size of tibial growth plates in AACs was enlarged and that the blood vessel density was also higher after exposure to the hypoxic environment for 14 days, while no such change was observed in TBCs. Altogether, our results revealed that the hypoxic environment has a potentially new role in increasing the blood vessel density of proximal tibial growth plates to strengthen and enhance the size of the growth plates, which may provide new insights for the therapeutic manipulation of hypoxia in poultry TD. PMID:28282429

Density and composition of microorganisms during long-term (418 day) growth of potato using biologically reclaimed nutrients from inedible plant biomass

NASA Technical Reports Server (NTRS)

Garland, J. L.; Cook, K. L.; Johnson, M.; Sumner, R.; Fields, N.; Sager, J. C. (Principal Investigator)

1997-01-01

A study evaluating alternative methods for long term operation of biomass production systems was recently completed at the Kennedy Space Center (KSC). The 418-day study evaluated repeated batch versus mixed-aged production of potato grown on either standard 1/2-strength Hoagland's nutrient solution or solutions including nutrients recycled from inedible plant material. The long term effects of closure and recycling on microbial dynamics were evaluated by monitoring the microbial communities associated with various habitats within the plant growth system (i.e., plant roots, nutrient solution, biofilms within the hydroponic systems, atmosphere, and atmospheric condensate). Plate count methods were used to enumerate and characterize microorganisms. Microscopic staining methods were used to estunate total cell densities. The primary finding was that the density and composition of microbial communities associated with controlled environmental plant growth systems are stable during long term operation. Continuous production resulted in slightly greater stability. Nutrient recycling, despite the addition of soluble organic material from the waste processing system, did not significantly increase microbial density in any of the habitats.

Density and composition of microorganisms during long-term (418 day) growth of potato using biologically reclaimed nutrients from inedible plant biomass

NASA Astrophysics Data System (ADS)

Garland, J. L.; Cook, K. L.; Johnson, M.; Sumner, R.; Fields, N.

1997-01-01

A study evaluating alternative methods for long term operation of biomass production systems was recently completed at the Kennedy Space Center (KSC). The 418-day study evaluated repeated batch versus mixed-aged production of potato grown on either standard 1/2-strength Hoagland's nutrient solution or solutions including nutrients recycled from inedible plant material. The long term effects of closure and recycling on microbial dynamics were evaluated by monitoring the microbial communities associated with various habitats within the plant growth system (i.e., plant roots, nutrient solution, biofilms within the hydroponic systems, atmosphere, and atmospheric condensate). Plate count methods were used to enumerate and characterize microorganisms. Microscopic staining methods were used to estimate total cell densities. The primary finding was that the density and composition of microbial communities associated with controlled environmental plant growth systems are stable during long term operation. Continuous production resulted in slightly greater stability. Nutrient recycling, despite the addition of soluble organic material from the waste processing system, did not significantly increase microbial density in any of the habitats.

Density and composition of microorganisms during long-term (418 day) growth of potato using biologically reclaimed nutrients from inedible plant biomass

NASA Astrophysics Data System (ADS)

1997-01-01

A study evaluating alternative methods for long term operation of biomass production systems was recently completed at the Kennedy Space Center (KSC). The 418-day study evaluated repeated batch versus mixed-aged production of potato grown on either standard 12-strength Hoagland's nutrient solution or solutions including nutrients recycled from inedible plant material. The long term effects of closure and recycling on microbial dynamics were evaluated by monitoring the microbial communities associated with various habitats within the plant growth system (i.e., plant roots, nutrient solution, biofilms within the hydroponic systems, atmosphere, and atmospheric condensate). Plate count methods were used to enumerate and characterize microorganisms. Microscopic staining methods were used to estimate total cell densities. The primary finding was that the density and composition of microbial communities associated with controlled environmental plant growth systems are stable during long term operation. Continuous production resulted in slightly greater stability. Nutrient recycling, despite the addition of soluble organic material from the waste processing system, did not significantly increase microbial density in any of the habitats.

Characterisation of structure-borne sound source using reception plate method.

PubMed

Putra, A; Saari, N F; Bakri, H; Ramlan, R; Dan, R M

2013-01-01

A laboratory-based experiment procedure of reception plate method for structure-borne sound source characterisation is reported in this paper. The method uses the assumption that the input power from the source installed on the plate is equal to the power dissipated by the plate. In this experiment, rectangular plates having high and low mobility relative to that of the source were used as the reception plates and a small electric fan motor was acting as the structure-borne source. The data representing the source characteristics, namely, the free velocity and the source mobility, were obtained and compared with those from direct measurement. Assumptions and constraints employing this method are discussed.

ATP as a biomarker of viable microorganisms in clean-room facilities

NASA Technical Reports Server (NTRS)

Venkateswaran, Kasthuri; Hattori, Noriaki; La Duc, Myron T.; Kern, Roger

2003-01-01

A new firefly luciferase bioluminescence assay method that differentiates free extracellular ATP (dead cells, etc.) from intracellular ATP (viable microbes) was used to determine the viable microbial cleanliness of various clean-room facilities. For comparison, samples were taken from both clean-rooms, where the air was filtered to remove particles >0.5 microm, and ordinary rooms with unfiltered air. The intracellular ATP was determined after enzymatically degrading the sample's free ATP. Also for comparison, cultivable microbial populations were counted on nutrient-rich trypticase soy agar (TSA) plates. Both the cultivable and ATP-based determinations indicate that the microbial burden was lower in clean-room facilities than in ordinary rooms. However, there was no direct correlation between the two sets of measurements because the two assays measured very different populations. A large fraction of the samples yielded no colony formers on TSA, but were positive for intracellular ATP. Subsequently, genomic DNA was isolated directly from selected samples and 16S rDNA fragments were cloned and sequenced, identifying nearest neighbors, many of which are known to be noncultivable in the media employed. It was concluded that viable microbial contamination can be reliably monitored by measurement of intracellular ATP, and that this method may be considered superior to cultivable colony counts due to its speed and its ability to report the presence of viable but noncultivable organisms. When the detection of nonviable microbes is of interest, the ATP assay can be supplemented with DNA analysis.

ATP as a biomarker of viable microorganisms in clean-room facilities.

PubMed

Venkateswaran, Kasthuri; Hattori, Noriaki; La Duc, Myron T; Kern, Roger

2003-03-01

A new firefly luciferase bioluminescence assay method that differentiates free extracellular ATP (dead cells, etc.) from intracellular ATP (viable microbes) was used to determine the viable microbial cleanliness of various clean-room facilities. For comparison, samples were taken from both clean-rooms, where the air was filtered to remove particles >0.5 microm, and ordinary rooms with unfiltered air. The intracellular ATP was determined after enzymatically degrading the sample's free ATP. Also for comparison, cultivable microbial populations were counted on nutrient-rich trypticase soy agar (TSA) plates. Both the cultivable and ATP-based determinations indicate that the microbial burden was lower in clean-room facilities than in ordinary rooms. However, there was no direct correlation between the two sets of measurements because the two assays measured very different populations. A large fraction of the samples yielded no colony formers on TSA, but were positive for intracellular ATP. Subsequently, genomic DNA was isolated directly from selected samples and 16S rDNA fragments were cloned and sequenced, identifying nearest neighbors, many of which are known to be noncultivable in the media employed. It was concluded that viable microbial contamination can be reliably monitored by measurement of intracellular ATP, and that this method may be considered superior to cultivable colony counts due to its speed and its ability to report the presence of viable but noncultivable organisms. When the detection of nonviable microbes is of interest, the ATP assay can be supplemented with DNA analysis.

Consecutive Plate Acoustic Suppressor Apparatus and Methods

NASA Technical Reports Server (NTRS)

Doychak, Joseph (Inventor); Parrott, Tony L. (Inventor)

1993-01-01

An apparatus and method for suppressing acoustic noise utilizes consecutive plates, closely spaced to each other so as to exploit dissipation associated with sound propagation in narrow channels to optimize the acoustic resistance at a liner surface. The closely spaced plates can be utilized as high temperature structural materials for jet engines by constructing the plates from composite materials. Geometries of the plates, such as plate depth, shape, thickness, inter-plate spacing, arrangement, etc., can be selected to achieve bulk material-like behavior.

Aseptic Laboratory Techniques: Plating Methods

PubMed Central

Sanders, Erin R.

2012-01-01

Microorganisms are present on all inanimate surfaces creating ubiquitous sources of possible contamination in the laboratory. Experimental success relies on the ability of a scientist to sterilize work surfaces and equipment as well as prevent contact of sterile instruments and solutions with non-sterile surfaces. Here we present the steps for several plating methods routinely used in the laboratory to isolate, propagate, or enumerate microorganisms such as bacteria and phage. All five methods incorporate aseptic technique, or procedures that maintain the sterility of experimental materials. Procedures described include (1) streak-plating bacterial cultures to isolate single colonies, (2) pour-plating and (3) spread-plating to enumerate viable bacterial colonies, (4) soft agar overlays to isolate phage and enumerate plaques, and (5) replica-plating to transfer cells from one plate to another in an identical spatial pattern. These procedures can be performed at the laboratory bench, provided they involve non-pathogenic strains of microorganisms (Biosafety Level 1, BSL-1). If working with BSL-2 organisms, then these manipulations must take place in a biosafety cabinet. Consult the most current edition of the Biosafety in Microbiological and Biomedical Laboratories (BMBL) as well as Material Safety Data Sheets (MSDS) for Infectious Substances to determine the biohazard classification as well as the safety precautions and containment facilities required for the microorganism in question. Bacterial strains and phage stocks can be obtained from research investigators, companies, and collections maintained by particular organizations such as the American Type Culture Collection (ATCC). It is recommended that non-pathogenic strains be used when learning the various plating methods. By following the procedures described in this protocol, students should be able to: ● Perform plating procedures without contaminating media. ● Isolate single bacterial colonies by the streak-plating method. ● Use pour-plating and spread-plating methods to determine the concentration of bacteria. ● Perform soft agar overlays when working with phage. ● Transfer bacterial cells from one plate to another using the replica-plating procedure. ● Given an experimental task, select the appropriate plating method. PMID:22617405

Enrichment of Acinetobacter spp. from food samples.

PubMed

Carvalheira, Ana; Ferreira, Vânia; Silva, Joana; Teixeira, Paula

2016-05-01

Relatively little is known about the role of foods in the chain of transmission of acinetobacters and the occurrence of different Acinetobacter spp. in foods. Currently, there is no standard procedure to recover acinetobacters from food in order to gain insight into the food-related ecology and epidemiology of acinetobacters. This study aimed to assess whether enrichment in Dijkshoorn enrichment medium followed by plating in CHROMagar™ Acinetobacter medium is a useful method for the isolation of Acinetobacter spp. from foods. Recovery of six Acinetobacter species from food spiked with these organisms was compared for two selective enrichment media (Baumann's enrichment and Dijkshoorn's enrichment). Significantly (p < 0.01) higher cell counts were obtained in Dijkshoorn's enrichment. Next, the Dijkshoorn's enrichment followed by direct plating on CHROMagar™ Acinetobacter was applied to detect Acinetobacter spp. in different foods. Fourteen different presumptive acinetobacters were recovered and assumed to represent nine different strains on the basis of REP-PCR typing. Eight of these strains were identified by rpoB gene analysis as belonging to the species Acinetobacter johnsonii, Acinetobacter calcoaceticus, Acinetobacter guillouiae and Acinetobacter gandensis. It was not possible to identify the species level of one strain which may suggests that it represents a distinct species. Copyright © 2015 Elsevier Ltd. All rights reserved.

Occurrence and characterization of Campylobacter in the Brazilian production and processing of broilers.

PubMed

Kuana, S L; Santos, L R; Rodrigues, L B; Borsoi, A; Moraes, H L S; Salle, C T P; Nascimento, V P

2008-12-01

Twenty-two commercial broiler flocks and their carcasses, totaling 546 samples (450 collected from a poultry farm and 96 from a slaughterhouse), were surveyed for the presence of Campylobacter. The positive results for Campylobacter among the analyzed samples were homogeneous, yielding 81.8% for cecal droppings, 80.9% for feces, and 80.4% for cloacal swabs. Pre-enrichment and direct plating showed that 77.85% and 81.8% of cloacal swabs, respectively, were positive for Campylobacter compared to 99.0% and 97.9% of carcasses testing positive with the pre-enrichment and direct plating methods. The Campylobacter count averaged 7.0 log10 colony-forming units (CFU)/g in cecal droppings, 5.15 log10 CFU/carcass after defeathering, and 4.24 log10 CFU/carcass after chilling. The samples were identified by the API Campy system as Campylobacter jejuni subsp. jejuni (68.8%), Campylobacter coli (8.3%), Campylobacter jejuni subsp. doylei (6.3%), Campylobacter upsaliensis (4.2%), and Campylobacter fetus subsp. fetus (2.1%). The analyzed broiler flocks were positive for Campylobacter in 81.8% of the cases, thus characterizing the occurrence of this pathogen in a broiler-producing region in southern Brazil. These results highlight the importance of programs targeted at the reduction of Campylobacter in poultry products, in order to minimize the risks for consumers.