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Survival of potentially pathogenic human-associated bacteria in the rhizosphere of hydroponically grown wheat.

Plants may serve as reservoirs for human-associated bacteria (H-AB) in long-term space missions containing bioregenerative life support systems. The current study examined the abilities of five human-associated potential pathogens, Pseudomonas aeruginosa, Pseudomonas cepacia, Staphylococcus aureus, Streptococcus pyogenes and Escherichia coli, to colonize and grow in the rhizosphere of hydroponically grown wheat, a candidate crop for life support. All of these bacteria have been recovered from past NASA missions and present potential problems for future missions. The abilities of these organisms to adhere to the roots of axenic five-day-old wheat (Triticum aestivum L. cv. Yecora rojo) were evaluated by enumeration of the attached organisms after a one hour incubation of roots in a suspension (approximately 10(8) cfu ml-1) of the H-AB. Results showed that a greater percentage of P. aeruginosa cells adhered to the wheat roots than the other four H-AB. Similarly incubated seedlings were also grown under attempted axenic conditions for seven days to examine the potential of each organism to proliferate in the rhizosphere (root colonization capacity). P. cepacia and P. aerogiunosa showed considerable growth, E. coli and S. aureus showed no significant growth, and S. pyogenes died off in the wheat rhizosphere. Studies examining the effects of competition on the survival of these microorganisms indicated that P. aeruginosa was the only organism that survived in the rhizosphere of hydroponically grown wheat in the presence of different levels of microbial competition.

Antibiosis↗

Effect of hydrogen peroxide treatment on microbial quality and appearance of whole and fresh-cut melons contaminated with Salmonella spp.

The efficacy of hydrogen peroxide treatment on the inactivation of Salmonella spp. inoculated on the external surface of cantaloupe and honeydew melon was investigated. Salmonella was inoculated onto whole cantaloupe and honeydew melon to a final concentration of 4.65 log(10) CFU/cm(2) and 3.13 log(10) CFU/g, respectively. Inoculated whole melons stored at 5 degrees C for up to 7 days were washed with water, 2.5% and 5% hydrogen peroxide at day 0 and 5. Hydrogen peroxide (2.5% and 5%) treatments of whole melon for 5 min caused a 3 log(10) CFU/cm(2) reduction of the indigenous surface microflora and a 3.0 log(10) CFU/cm(2) reduction in Salmonella spp. on all melon surfaces. The efficacy of the hydrogen peroxide treatments was less when the interval between inoculation and treatment of cantaloupe exceeded 24 h. Unlike cantaloupe fresh-cut pieces, Salmonella was not recovered from fresh-cut pieces prepared from treated whole honeydew melon. Growth of Salmonella occurred in cantaloupe fresh-cut pieces stored at 10 or 20 degrees C, and by 2 weeks, levels reached approximately 1 log CFU/g. A rapid decline in appearance and overall acceptability was observed in fresh-cut pieces prepared from untreated whole cantaloupe. While Salmonella was recovered from fresh-cut pieces from and whole treated cantaloupe, sanitizing the surface of contaminated whole melons with hydrogen peroxide before and after cutting and storage of the fresh-cut pieces at 5 degrees C can enhance the microbial safety and acceptability rating for about 2 weeks after processing.

Anti-Infective Agents, Local↗

Identification of probiotic microorganisms in South African products using PCR-based DGGE analysis.

Probiotic microorganisms in commercial yoghurts and other food products are currently identified by traditional methods such as growth on selective media, morphological and biochemical characteristics. In this study, PCR-based DGGE analysis was used for the rapid and accurate identification of probiotic microorganisms from South African yoghurts and lyophilized preparations in capsule and tablet form. To identify the microorganisms present in these products, the DGGE profiles obtained were compared to two reference markers (A and B) composed of five lactobacilli and seven Bifidobacterium species, respectively. The results obtained were confirmed by species-specific PCR, as well as sequence analyses of unknown bands not present in the reference markers. It was found that only 54.5% of the probiotic yoghurts contained the microorganisms stated on the label compared to only a third (33.3%) of the lyophilized probiotic products. Some Bifidobacterium species were incorrectly identified and various microorganisms were detected that were not listed on the label. Sequence analyses confirmed the presence of Streptococcus spp. other than the yoghurt starter, Streptococcus thermophilus, in some of these products and in some instances label information was vague and non-scientific. PCR-based DGGE analyses proved to be a valuable culture-independent approach for the rapid and specific identification of the microbial species present in South African probiotic products.

Bifidobacterium↗

The effect of reuterin on the lag time of single cells of Listeria innocua grown on a solid agar surface at different pH and NaCl concentrations.

The lag time of single cells of Listeria innocua grown on the surface of Brain Heart Infusion Agar was studied by microscopy and image analysis. An experimental set-up that enabled relocation of the cells on the agar surface was developed and used to collect data from 50 to 100 individual cells at a time. Reuterin was added at different concentrations (0-10 AU/ml) and it was observed that it increased both the lag time of the cells and its variance. Furthermore, for a large proportion of cells, reuterin completely prevented the cell division within the time of observation. Reuterin in combination with low pH inhibited the cell division even more efficiently. A similar effect was observed for the combination of reuterin and sodium chloride. Our experimental set-up provides a good model system for generating data on the lag time of single cells on solid surfaces, which can improve the predictions of microbial growth on solid food matrices.

Agar↗

Dynamic computer simulation of Clostridium perfringens growth in cooked ground beef.

The objective of this study was to develop a computer simulation algorithm to dynamically estimate and predict the growth of Clostridium perfringens in cooked ground beef. The computational algorithm was based on the implicit form of the Gompertz model, the growth kinetics of C. perfringens in cooked ground beef, and the fourth-order Runge-Kutta numerical method. This algorithm was validated using a cocktail of three strains of C. perfringens spores grown under isothermal, square-waved, linear cooling, and exponential cooling temperature profiles. In general, the results of computer simulation matched closely with the experimental data with the absolute errors less than 0.5 log(10) CFU/g. This method may be a useful tool for the food industry, regulatory agencies, distributors, and retailers to predict the effect of temperature abuse on the microbial safety of C. perfringens and other foodborne pathogens in processed meat products.

Animals↗

A novel modelling approach for predicting microbial growth in a raw cured meat product stored at 3 degrees C and at 12 degrees C in air.

To predict microbial growth during chill storage of a traditional Greek raw sausage, a numerical model was developed and validated. In our novel approach, the specific growth rate of each microbial population was calculated on the basis of the main microbial populations grown in the sausage. In addition, the specific destructive effect of the sausage ecosystem was introduced to evaluate microbial growth. The model was integrated by the Runge-Kutta method and the parameter values were optimised by the least squares method. Fitting of the model to the experimental data derived from four sausage batches stored aerobically at 3 and 12 degrees C successfully described the microbial growth kinetics in the sausage niche. Finally, the parameter values estimated by the fitting of the model on the data set from each batch were used to predict microbial growth in the other batches at both storage temperatures.

Bacteria↗

Ecology of mixed biofilms subjected daily to a chlorinated alkaline solution: spatial distribution of bacterial species suggests a protective effect of one species to another.

Three bacterial strains (Kocuria sp. C714.1, Brevibacterium linens B337.1 and Staphylococcus sciuri CCL101) were grown together on stainless steel and were subjected daily to a commercial alkaline chlorine solution (22 mg l-1 of free chlorine, pH 11) over a period of 4 weeks. After the daily chemical shock, culture madia [1:20 dilution of tryptic soy broth (TSB-YE/20) or diluted whey] was deposited on the biofilms. The chemical shocks led first to a drop in the culturable population, followed by an increase and finally stabilization at around 106-107 CFU cm-2 by day 11 of the experiment. These changes in the microbial population can be attributed to a decreasing susceptibility to the antimicrobial agent with biofilm age, and to the consumption of free chlorine by biofilm exoproteins. The microbial composition appeared to be linked to the free chlorine concentration that depended on exoprotein production. At the end of the experiment, exoprotein production was greater for biofilms grown in TSBYE/20 than in whey. As a consequence, biofilms grown in whey did not neutralize the chlorine and the dominant strain was the one having the highest resistance to chlorine: K. varians. When biofilm were grown in TSBYE/20, chlorine was neutralized and the dominant strain was the one having the highest growth rate: S. sciuri. The presence of chlorine may also explain the distribution of S. sciuri cells as a ring around Kocuria sp. microcolonies. When chlorine was totally consumed by the biofilm during the chemical shock, S. sciuri was no longer grouped around Kocuria sp. microcolonies but was evenly scattered over the substratum as single cells or in small clusters, as it was before any chemical treatment. These findings strongly suggest protection of S. sciuri by Kocuria sp. microcolonies against the chlorinated solution. This phenomenon, added to the low susceptibility phenotype of the biofilm cells, could at least partly explain the survival of microbial cells in an adverse environment.

Anti-Bacterial Agents↗

Use of microcosms to determine persistence of Escherichia coli in recreational coastal water and sediment and validation with in situ measurements.

AIMS: To determine the persistence of the faecal indicator organism Escherichia coli in recreational coastal water and sediment using laboratory-based microcosms and validation with in situ measurements. METHODS AND RESULTS: Intact sediment cores were taken from three distinct coastal sites. Overlying estuarine water was inoculated with known concentrations of E. coli and decay rates from both overlying water and sediment were determined following enumeration by the membrane filtration method at fixed time intervals over a 28-day period. It was demonstrated that E. coli may persist in coastal sediment for >28 days when incubated at 10 degrees C. Escherichia coli survival was found to have an inverse relationship with temperature in both water and sediment. In general the decay rate for E. coli was greater in water than in sediment. Small particle size and high organic carbon content were found to enhance E. coli survival in coastal sediments in the microcosms. CONCLUSIONS: Results of this microcosm study demonstrated the more prolonged survival of E. coli in coastal sediments compared with overlying water, which may imply an increased risk of exposure because of the possible resuspension of pathogenic micro-organisms during natural turbulence or human recreational activity. SIGNIFICANCE AND IMPACT OF THE STUDY: A more accurate estimate of exposure risk has been described which may subsequently be used in a quantitative microbial risk assessment for recreational coastal waters.

Bacteriological Techniques↗

Quantification of mcrA by quantitative fluorescent PCR in sediments from methane seep of the Nankai Trough.

A quantitative fluorogenic PCR method for group-specific methyl coenzyme M reductase subunit A genes (mcrA) from methanotrophic archaea was established and applied to the characterization of microbial communities in anoxic methane seep sediments at the accretionary prism of the Nankai Trough. All of the previously identified subgroups of anaerobic methanotroph (ANME) mcrA genes were detected in the cores up to 25 cm below the seafloor, but distributional patterns of mcrA genes were found to differ according to depth. These findings suggest a distinct distribution of phylogenetically and physiologically diverse methanotrophic archaea that mediate methane oxidation in the anoxic sediments. This quantification method will contribute to future investigations of methanotrophic microbial ecosystems in anoxic marine sediments.

Anaerobiosis↗

Bacterial community dynamics and polycyclic aromatic hydrocarbon degradation during bioremediation of heavily creosote-contaminated soil.

Bacterial community dynamics and biodegradation processes were examined in a highly creosote-contaminated soil undergoing a range of laboratory-based bioremediation treatments. The dynamics of the eubacterial community, the number of heterotrophs and polycyclic aromatic hydrocarbon (PAH) degraders, and the total petroleum hydrocarbon (TPH) and PAH concentrations were monitored during the bioremediation process. TPH and PAHs were significantly degraded in all treatments (72 to 79% and 83 to 87%, respectively), and the biodegradation values were higher when nutrients were not added, especially for benzo(a)anthracene and chrysene. The moisture content and aeration were determined to be the key factors associated with PAH bioremediation. Neither biosurfactant addition, bioaugmentation, nor ferric octate addition led to differences in PAH or TPH biodegradation compared to biodegradation with nutrient treatment. All treatments resulted in a high first-order degradation rate during the first 45 days, which was markedly reduced after 90 days. A sharp increase in the size of the heterotrophic and PAH-degrading microbial populations was observed, which coincided with the highest rates of TPH and PAH biodegradation. At the end of the incubation period, PAH degraders were more prevalent in samples to which nutrients had not been added. Denaturing gradient gel electrophoresis analysis and principal-component analysis confirmed that there was a remarkable shift in the composition of the bacterial community due to both the biodegradation process and the addition of nutrients. At early stages of biodegradation, the alpha-Proteobacteria group (genera Sphingomonas and Azospirillum) was the dominant group in all treatments. At later stages, the gamma-Proteobacteria group (genus Xanthomonas), the alpha-Proteobacteria group (genus Sphingomonas), and the Cytophaga-Flexibacter-Bacteroides group (Bacteroidetes) were the dominant groups in the nonnutrient treatment, while the gamma-Proteobacteria group (genus Xathomonas), the beta-Proteobacteria group (genera Alcaligenes and Achromobacter), and the alpha-Proteobacteria group (genus Sphingomonas) were the dominant groups in the nutrient treatment. This study shows that specific bacterial phylotypes are associated both with different phases of PAH degradation and with nutrient addition in a preadapted PAH-contaminated soil. Our findings also suggest that there are complex interactions between bacterial species and medium conditions that influence the biodegradation capacity of the microbial communities involved in bioremediation processes.

Bacteria↗

A correlation study of inflammatory cell mobilization in response to subgingival microbial colonization.

This study evaluated site-by-site the relations between subgingival microbial colonization and gingival tissue reactions. Experimental, deep periodontal defects were established at buccal surfaces of mandibular and maxillary canine teeth in 5 beagle dogs. The root surfaces were instrumented by a flame-shaped, fine-grained, rotating diamond point, or by a sharp curet. Following a 10-day postsurgical healing period, the dogs were fed a plaque-inducing diet for 70 days. The animals were then sacrificed and tissue blocks of the experimental sites including teeth and periodontal tissues were secured. The buccal gingiva was removed and processed for histomorphometric analysis while the teeth were prepared for scanning electron microscopic evaluation of the extent of subgingival microbial colonization. The results revealed that inflammatory cell density in the junctional epithelium and in the connective tissue were positively correlated to subgingival microbial colonization (P < 0.01). Furthermore, the degree of significance decreased with increasing distance from the plaque. The present study demonstrates that a close relation may exist between the extent of subgingival microbial colonization and inflammatory gingival tissue reactions.

Animals↗

Response of Salmonella and Escherichia coli O157:H7 to UV energy.

To determine the efficacy of a UV light treatment at 253.7 nm (UVC light) on microbial growth, plates containing tryptic soy agar plus 50 ppm of nalidixic acid (TSAN) were inoculated with known concentrations of five-strain cocktails of Salmonella and Escherichia coli O157:H7 and subjected to different UVC treatments. The concentration of the cocktail inoculum was determined with TSAN prior to inoculation. Serial dilutions were carried out, and inoculation levels of 10(0) to 10(8) CFU/ ml were tested for each pathogen. Multiple replications of doses of UV light ranging from 1.5 to 30 mW/cm2 were applied to different cocktail concentrations, and doses of > 8.4 mW/cm2 resulted in a 5-log reduction of Escherichia coli O157:H7, while a 5-log reduction of Salmonella was observed with doses of > 14.5 mW/cm2. Results for both organisms yielded sigmoidal inactivation curves. UVC light is effective in reducing microbial populations of pathogens on agar surfaces.

Colony Count, Microbial↗

Comparison of microbial isolates and susceptibility patterns from the external ear canal of dogs with otitis externa.

Otic exudate was obtained from 33 dogs with otitis externa for cytopathology and culture. Two samples were taken from the same location in the external ear canal, for a total of 100 samples. Thirty-six (36%) samples isolated only a single organism, of which 21 (21%) were Malassezia spp. Two organisms were present in 23 (23%) of the samples. Cultures of the two samples agreed in 40 (80%) of the 50 pairs. Cytopathology agreed with culture results only 68% of the time. Cytopathology and culture may not be as definitive as previously assumed because of apparent variability of the microbial population within the external canal.

Animals↗

[The frequency of staphylococcal colonization of the intestines in children with the manifestations of dysbacteriosis].

In 2100 children of different age groups the microbiocenosis of the large intestine was studied. The study revealed that the colonization of the mucous membrane of the large intestine with staphylococci developed in 30% of children with intestinal dysbacteriosis. Young children were mainly affected (91%). The prevailing species among isolated staphylococci was S. aureus (86%), capable of persistence in the intestine (30.9%). In children non typing S. aureus strains mainly circulated (70%), and among phage-typing strains isolates of phage group III prevailed (70.2%). The colonization of the intestine with coagulase-negative staphylococci was possible (14%). Microecological intestinal disturbances in children of different age groups were characterized by different degrees of changes in normal microflora with the prevalence of opportunistic microorganisms in the microbial picture.

Bacteriophage Typing↗

Inhibitory effects of spices and herbs towards Shigella sonnei and S. flexneri.

The use of spices and herbs, their essential oils or their active compounds as means of control of pathogens constitutes an alternative to chemical additives In the present study the antibacterial activities of cloves, thyme, oregano, rosemary and basil on Shigella have been established. Although in a model system in agar media addition of 1% basil could contribute to the 'hurdle' principle and delay outgrowth of Shigella sp. this was not confirmed in real food conditions: the presence of 1% basil did not affect growth of Shigella in potato puree at 22 degrees C or survival at 7 degrees C in spaghetti sauce. Thyme and essential oils and thymol and carvacrol showed inhibition of Shigella sp. in the agar well diffusion method (MIC 0.1-1.0%) and they have potential to be used as a desinfectant in the washing water e.g. in the process line of minimal processed vegetables. However, more studies combining sensoric properties with microbial analysis are needed to investigated the possible use of these compounds.

Colony Count, Microbial↗

[Microbiological diagnosis of lower respiratory tract infections].

Successful microbiological diagnosis of lower respiratory tract infections demands a close cooperation between clinician and clinical microbiologist. Because of the broad spectrum of possible respiratory pathogens precise requests are necessary for adequate laboratory procedures. The high rate of potential pathogens requires quantitative microbiological and cytological data in order to differentiate between colonisation and infection. Pathophysiological reactions on microbial colonisation of the bronchial tree may contribute to acute exacerbations of a chronic bronchitis. The precise role of microbial nocuous agents, however, remains to be clarified.

Bronchi↗

[Characteristics of the transphase and transovarial transmission of Rickettsia sibirica by ixodid and argasid ticks].

Both the specific vector Dermacentor silvarum and the nonspecific one Ornithodoros papillipes are shown to become Rickettsia sibirica infected while sucking blood on the infected guinea pigs. Meanwhile, the specific vector D. silvarum is more sensitive to R. sibirica than the nonspecific. O papillipes. The infectivity of D. silvarum and O. papillipes ticks by R. sibirica was found to be dependent on the intensity of guinea pigs' rickettsemia. Transovarial transmission of Rickettsia has been established for the D. silvarum and O. papillipes ticks, but the Rickettsia titer in the infested D. silvarum larvae exceeded that O. papillipes. Survival of D. silvarum ticks infected by various concentrations of the R. sibirica microbial bodies was found to exceed that of O. papillipes.

Animals↗

Drinking-water quality and issues associated with water vending machines in the city of Los Angeles.

The study reported in this paper sampled 40 water vending machines distributed throughout the city of Los Angeles, California, in the water servicing area of the Los Angeles Department of Water and Power. The authors examined water samples for temperature, turbidity, chlorine content, fungal growth, coliform bacteria, fecal coliform bacteria, Pseudomonas spp., Pseudomonas aeruginosa, and heterotrophic plate counts using plate count agar and R2A agar. Roughly 48 percent of the original 40 samples were sampled in a second or third round, and these subsequent samples were examined for coliform bacteria, fecal coliform bacteria, Pseudomonas spp., Pseudomonas aeruginosa, and both heterotrophic plate counts. Observations were made of vending-machine external conditions, identifying information, and permits. Statistically significant associations were found between operator accessibility and poor machine conditions, operator accessibility and permits, and operator accessibility and the servicing interval. Statistically significant associations also were found between operator accessibility and presence of fungi, Pseudomonas spp., and Pseudomonas aeruginosa, and between presence of fungi and the servicing interval. In addition, statistically significant associations indicate that the quality of machine maintenance is a factor in microbial levels. The microbial content of the water suggested a need for further research into potential health effects in susceptible populations.

Beverages↗