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Effects of form of the diet on anatomical, microbial, and fermentative development of the rumen of neonatal calves.

Eight neonatal, Holstein bull calves were paired by birth date and birth weight and randomly assigned to either a finely ground or unground control diet (chopped hay and rolled grain) to study the effects of the physical form of the diet on anatomical, microbial, and fermentative development of the rumen. The diets varied in particle size but were identical in composition (25% alfalfa hay and 75% grain mix). Calves were fed milk at 8% of birth weight daily until weaning. Feed intake was equalized for each pair of calves. Ruminal fluid samples were collected from ruminal cannulas to determine pH, fermentation products, and buffering capacity and to enumerate bacteria. Calves were slaughtered at 10 wk of age, and weights of the full and empty reticulorumen, abomasum, and omasum were recorded. Ruminal tissue samples were taken to assess papillary development by morphometric measurements. Calves had similar body weights at wk 10. Ruminal pH was affected by age and was lower for calves fed the ground diet. Total anaerobic bacterial counts were not affected by the physical form of the diet; however, calves fed the ground diet had lower numbers of cellulolytic bacteria and higher numbers of amylolytic bacteria than did calves fed the unground diet. Physical form of the diet did not affect the weights of the reticulorumen whether full or empty. However, calves fed the ground diet had heavier omasum weights, both full and empty. Physical form of the diet affected papillary size and shape but did not influence the muscle thickness of rumen. Results indicated that the physical form of the diet had a significant influence on the anatomical and microbial development of the forestomac and, therefore, might influence future performance.

Animal Feed↗

Effects of carbon dioxide on bacterial growth parameters in milk as measured by conductivity.

Inhibition of bacterial growth by dissolved carbon dioxide (CO2) has been well established in many foods including dairy foods. However, the effects of dissolved CO2 on specific growth parameters such as length of lag phase, time to maximum growth rate, and numbers of organisms at the stationary phase have not been quantified for organisms of concern in milk. The effect of dissolved CO2 concentrations of 0.6 to 61.4 mM on specific bacterial growth parameters in raw or single organism inoculated sterile milk was determined at 15 degrees C by conductance. Commingled raw or sterile milks were amended to a final concentration of 0.5 mg/ml each of urea and arginine HCl. Sterile milks were inoculated singly with one of six different microorganisms to a final concentration of approximately 10(2) to 10(3) cfu/ml; raw milk was adjusted to a final indigenous bacterial population of approximately 10(3) cfu/ml. Conductivity of the milk was recorded every 60 s over 4 to 5 d in a circulating apparatus at 15 degrees C. Conductivity values were fit to Gompertz equations and growth parameters calculated. Conductance correlated with plate counts and was satisfactory for monitoring microbial growth. Data fit the Gompertz equation with high correlation (R2 = 0.96 to 1.00). In all cases, dissolved CO2 significantly inhibited growth of raw milk bacteria, influencing lag, exponential, and stationary growth phases as well as all tested monocultures.

Animals↗

Application of laser scanning for the rapid and automated detection of bacteria in water samples.

It is widely accepted that the heterotrophic plate count method may not support the growth of all viable bacteria which may be present within a water sample and that alternative procedures using 'viability markers' may yield additional information. In this study, ChemChrome B (CB), which is converted to a fluorescent product by esterase activity, was used to stain viable bacteria (captured by membrane filtration) from potable water samples. The labelled bacteria from each sample were subsequently enumerated using a novel laser scanning instrument (ChemScan). Analysis of 107 potable water samples using this procedure demonstrated the presence of a significantly greater number of bacteria than were detected by culture (z-test, P < 0.05). The mean number of bacteria isolated by culture on R2A agar incubated at 22 degrees C for 7 d was only 25.2% of the total number of viable bacteria detected using the CB/ChemScan viability assay. Further analysis of 81 water samples using a 5-cyano-2,3,4-tolyl-tetrazolium chloride (CTC) viability assay also demonstrated the presence of many viable bacteria which were not capable of growth under the culture conditions employed in this study. However, the results indicate that ChemChrome B has the ability to stain a significantly greater number of heterotrophs than CTC (z-test, P < 0.05). In contrast, six potable waters were identified in which the CTC viability assay resulted in counts greater than those obtained using CB. The ChemScan instrument was successfully used for rapid and accurate enumeration of labelled micro-organisms, allowing information on the total viable microbial load of a water sample to be determined within 1 h. Furthermore, the ChemScan system has the potential for use in detecting specific organisms labelled with fluorescently-labelled antibodies or nucleic acid probes.

Bacteria↗

Biological potential of extraterrestrial materials. 2. Microbial and plant responses to nutrients in the Murchison carbonaceous meteorite.

Meteoritic materials are investigated as potential early planetary nutrients. Aqueous extracts of the Murchison C2 carbonaceous meteorite are utilized as a sole carbon source by microorganisms, as demonstrated by the genetically modified Pseudomonas fluorescence equipped with the lux gene. Nutrient effects are observed also with the soil microorganisms Nocardia asteroides and Arthrobacter pascens that reach populations up to 5 x 10(7) CFU/ml in meteorite extracts, similar to populations in terrestrial soil extracts. Plant tissue cultures of Asparagus officinalis and Solanum tuberosum (potato) exhibit enhanced pigmentation and some enhanced growth when meteorite extracts are added to partial nutrient media, but inhibited growth when added to full nutrient solution. The meteorite extracts lead to large increases in S, Ca, Mg, and Fe plant tissue contents as shown by X-ray fluorescence, while P, K, and Cl contents show mixed effects. In both microbiological and plant tissue experiments, the nutrient and inhibitory effects appear to be best balanced for growth at about 1:20 (extracted solid : H2O) ratios. The results suggest that solutions in cavities in meteorites can provide efficient concentrated biogenic and early nutrient environments, including high phosphate levels, which may be the limiting nutrient. The results also suggest that carbonaceous asteroid resources can sustain soil microbial activity and provide essential macronutrients for future space-based ecosystems.

Arthrobacter↗

The Freter model: a simple model of biofilm formation.

A simple, conceptual model of biofilm formation, due to R. Freter et al. (1983), is studied analytically and numerically in both CSTR and PFR. Two steady state regimes are identified, namely, the complete washout of the microbes from the reactor and the successful colonization of both the wall and bulk fluid. One of these is stable for any particular set of parameter values and sharp and explicit conditions are given for the stability of each. The effects of adding an anti-microbial agent to the CSTR are examined.

Algorithms↗

Luminometric measurement of population activity of genetically modified Pseudomonas fluorescens in the soil.

Genetically modified cells of Pseudomonas fluorescens, chromosomally marked with genes for bioluminescence, were inoculated into sterile soil microcosms. During incubation for 90 days, viable cell concentration did not change significantly but light output, measured by luminometry, decreased, indicating reduced metabolic activity due to lack of substrates. Amendment with nutrients resulted in parallel increases in both luminescence and dehydrogenase activity. Luminometry therefore enables rapid monitoring of the activity of populations of luminescence-marked microbial inocula in the soil, with greater sensitivity and selectivity than traditional techniques.

Bacteriological Techniques↗

Characterisation of the dominant bacterial population in modified atmosphere packaged farmed halibut (Hippoglossus hippoglossus) based on 16S rDNA-DGGE.

It is not well understood why Atlantic halibut (Hippoglossus hippoglossus) has longer shelf-life than most other white fish species. Our approach was to examine the microbiological diversity of the spoilage microbiota during modified atmosphere (MA) packaging of farmed Atlantic halibut. Portions were packaged with gas mixtures of CO(2):N(2) and CO(2):O(2) (50%:50%) and with air as a reference. The packages were stored at 4 degrees C and samples were taken 6 times during the 23 days of storage. Analyses with molecular techniques (PCR-DGGE) determined profiles of the bacterial populations in the various samples and sequencing detected the bacterial species present. In addition, samples were analysed for microbial, chemical and sensory parameters. The shelf-life was 10-13 days when stored in air and between 13 and 20 days for MA packages, with oxygen-enriched packages suggested as the better gas mixture, based on microbial growth and sensory scores. From sequence analyses of the bacterial population Photobacterium phosphoreum and Pseudomonas spp. were found to dominate in the halibut. Brochothrix thermosphacta was found in most samples at the end of the storage period. Shewanella putrefaciens was found sporadically and in low concentrations based on microbial methods, but not detected by PCR-DGGE.

Animals↗

Pesticides as a source of microbial contamination of salad vegetables.

Ten commercially available pesticides (insecticides, herbicides and fungicides), used during the production of vegetable produce, were examined as potential sources of microbial contaminants. As purchased, none of the pesticides showed the presence of viable microorganisms (< 5 CFU/ml). Using an agar plate diffusion assay, they did not inhibit a range of bacteria of spoilage and public health significance on vegetable produce. After reconstitution in sterile water to their recommended concentration, two of the pesticides supported the survival and growth of inoculated species of Pseudomonas, Salmonella and Escherichia coli. Listeria monocytogenes did not survive after inoculation into any of the pesticides. Pesticides were reconstituted in different sources of agricultural water (bore, dam and river) and examined for survival and growth of microorganisms naturally present in these waters. On storage at 30 degrees C for 48 h, nine of the pesticides supported the growth of bacterial species present in these waters. Predominant species in the pesticide solutions, before and after storage, varied according to the source, but species of Pseudomonas, Acinetobacter and Aeromonas and various coliforms exhibited significant growth. Unless managed properly (reconstituted in potable water, and used without lengthy storage), pesticides could contribute to the microbial load of vegetable produce, thereby affecting their shelf-life and public health safety.

Bacteria↗

Use of hydrogen peroxide in combination with nisin, sodium lactate and citric acid for reducing transfer of bacterial pathogens from whole melon surfaces to fresh-cut pieces.

Hydrogen peroxide (2.5%) alone or hydrogen peroxide (1%) in combination with nisin (25 microg/ml), sodium lactate (1%), and citric acid (0.5%) (HPLNC) were investigated as potential sanitizers for reducing Escherichia coli O157:H7 or Listeria monocytogenes populations on whole cantaloupe and honeydew melons. Whole cantaloupes inoculated with E. coli O157:H7 and L. monocytogenes at 5.27 and 4.07 log10 CFU/cm2, respectively, and whole honeydew melons inoculated with E. coli O157:H7 and L. monocytogenes at 3.45 and 3.05 log10 CFU/cm2, respectively, were stored at 5 degrees C for 7 days. Antimicrobial washing treatments were applied to inoculated whole melons on days 0 or 7 of storage and surviving bacterial populations and the numbers transferred to fresh-cut pieces were determined. At days 0 and 7 treatment with HPLNC significantly (p<0.05) reduced the numbers of both pathogens, by 3 to 4 log CFU/cm2 on both types of whole melon. Treatment with HPLNC was significantly (p<0.05) more effective than treatment with 2.5% hydrogen peroxide. While fresh-cut pieces prepared from stored whole melons were negative for the pathogens by both direct plating and by enrichment, fresh-cut pieces from cantaloupe melons treated with 2.5% hydrogen peroxide were positive for both pathogens and pieces from honeydew melons were positive for E. coli 0157:H7. The native microflora on fresh-cut melons were also substantially reduced by HPLNC treatment of whole melons. The results suggest that HPLNC could be used to decontaminate whole melon surfaces and so improve the microbial safety and quality of fresh-cut melons.

Anti-Bacterial Agents↗

Effect of gamma irradiation on viability and DNA of Staphylococcus epidermidis and Escherichia coli.

Gamma irradiation is widely used for sterilization; however, its effect on elimination of amplifiable DNA, an issue of relevance to molecular diagnostic approaches, has not been well studied. The effect of gamma irradiation on the viability of Staphylococcus epidermidis and Escherichia coli (using quantitative cultures) and on their DNA (using quantitative 16S rRNA gene PCR) was evaluated. Viability was abrogated at 2.8 and 3.6 kGy for S. epidermidis and E. coli, respectively. The radiation dose required to reduce viable bacteria by one log10 (D10 value) was 0.31 and 0.35 kGy for S. epidermidis and E. coli, respectively. D10 values for amplifiable DNA extracted from bacteria were 2.58 and 3.09 kGy for S. epidermidis and E. coli, respectively, whereas D10 values for amplifiable DNA were significantly higher for DNA extracted from irradiated viable bacterial cells (22.9 and 52.6 kGy for S. epidermidis and E. coli, respectively; P<0.001). This study showed that gamma irradiation of DNA in viable bacterial cells has little effect on amplifiable DNA, was not able to eliminate amplifiable 16S rRNA genes at a dose of up to 12 kGy and cannot therefore be used for elimination of DNA contamination of PCR reaction components or laboratory equipment when this DNA is present in microbial cells. This finding has practical implications for those using molecular diagnostic techniques in microbiology.

Colony Count, Microbial↗

Microbial colonization of the Groningen speaking valve and its relationship to valve failure.

Speaking valve failure may be related to biofilm development. An inter-observer validated method of assessing microbial overgrowth in defective prostheses was developed. Two independent observers recorded colonisation of the valves on a 100 mm linear analogue scale in several defined areas. Inter-observer agreement was high in all areas. In vitro opening pressure and derived forward resistances were measured. The relationship between colonization of the valves and the in vitro measurements was investigated. Significant correlations for a two-tailed Spearman's test were recorded between the resistance and microbial colonization of the hinge and oesophageal areas (rs 0.22: P < 0.05, rs 0.25: P < 0.05, n = 80). There was no significant association for resistance and colonization of the tracheal surface, and no significant association between the opening pressure and the colonization. This work suggests that microbial colonization of the prosthesis in the oesophageal and hinge areas is associated with valve failure.

Colony Count, Microbial↗

Occurrence of necrotizing enterocolitis may be dependent on patterns of bacterial adherence and intestinal colonization: studies in Caco-2 tissue culture and weanling rabbit models.

Necrotizing enterocolitis (NEC) is one of the leading causes of death in neonatal intensive care units. The underlying pathophysiology of NEC is poorly defined, although there is a suggestion that bacterial agents play an important role in the process. In this study, we evaluated bacterial isolates from 17 NEC cases and matched asymptomatic control infants. Isolates from NEC patients were no more likely than control isolates to be adherent to enterocytes, as assessed by a Caco-2 cell tissue culture model. Adherent Escherichia coli isolates, from both NEC cases and controls, were able to cause pathologic changes typical of NEC in a weanling rabbit ileal loop model. Adherence of E. coli strains to Caco-2 cells, and subsequent production of disease in weanling rabbits, could be blocked by coinfection with Gram-positive isolates from control children. In contrast, in three of four instances, adherent E. coli from NEC cases retained their adherence and caused illness in rabbits when coinfected with Gram-positive isolates from the homologous child. Our data suggest that patterns of intestinal adherence, as influenced by the underlying intestinal microbial ecology, play a role in the pathophysiology of NEC.

Animals↗

Characterization of microbial populations and volatile fatty acid concentrations in the jejunum, ileum, and cecum of pigs weaned at 17 vs 24 days of age.

In a series of five 17-d replicate trials, a total of 54 cannulated and 12 noncannulated pigs were used to determine the effects of weaning age (17 d or 24 d) on pH, dry matter percentage, aerobic and anaerobic microflora, lactate, and volatile fatty acid (VFA) concentrations in the jejunum, ileum, and cecum of weanling pigs. At -14 d of age, cannulated pigs were surgically fitted with T-cannulas in the jejunum (n = 20), ileum (n = 18), or cecum (n = 16). Upon weaning, cannulated pigs were individually caged in an environmentally controlled room with ad libitum access to a phase starter diet and water. Noncannulated pigs were killed at weaning and samples were collected from the jejunum, ileum, and cecum. Digesta and fecal swabs from cannulated pigs were collected twice weekly. The pH of cecal contents was lower (P < 0.05) and dry matter percentage was greater (P < 0.05) than those ofjejunal or ileal contents. Pigs weaned at 24 d of age had increased (P < 0.05) E. coli populations 3 d postweaning compared to preweaning populations, regardless of site of collection, whereas this increase was not observed in pigs weaned at 17 d of age. Unweaned pigs maintained higher (P < 0.05) lactobacilli populations compared to weaned pigs; however, populations declined (P < 0.05) in both groups by 3 d postweaning, with pigs weaned at 24 d of age having lactobacilli populations greater than pigs weaned at 17 d of age. Fecal populations of E. coli and lactobacilli declined (P < 0.05), whereas fecal bifidobacteria populations increased (P < 0.05) postweaning, regardless of weaning age. Concentrations of total fecal anaerobes declined (P < 0.05) in pigs weaned at 17 d of age but were maintained in pigs weaned at 24 d of age. Volatile fatty acid concentrations were greater (P < 0.05) in the cecum than in the jejunum or ileum, and acetic acid concentrations decreased (P < 0.05) postweaning regardless of weaning age. A tendency for L+ lactate concentrations to be greater (P < 0.07) in the ileum and jejunum vs the cecum was observed. Results indicate that weaning and weaning age have significant effects on microbial populations and VFA concentrations.

Age Factors↗

Application of predictive models to estimate Listeria monocytogenes growth on frankfurters treated with organic acid salts.

Organic acid salts including sodium lactate, sodium diacetate, potassium benzoate, potassium sorbate, and their combinations were assessed as potential inhibitors of Listeria monocytogenes growth on frankfurters. Predictive models for L. monocytogenes growth on frankfurters treated with these salts were compared to select a proper L. monocytogenes growth curve model under these conditions. Sigmoidal equations, including logistic and Gompertz equations, are widely used to describe bacterial growth. In this study, the reparameterized Gompertz model provided a better fit to the L. monocytogenes growth data compared with the other models that were included in this study. Rather than a fixed value for the maximum number of organisms, the reparameterized Gompertz model allows this quantity to be estimated from the data to determine the effect, if any, of the treatments on maximum population density. This information is expected to improve practical methodology for hazard characterization of microbial pathogens on ready-to-eat meat products.

Animals↗

Impact of commercial processing on the microbiology of shell eggs.

Shell egg microbiology has been studied extensively, but little information is available on how modern U.S. processing conditions impact microbial populations. As regulations are being drafted for the industry, such information can be important for determining processing steps critical to product safety. Five different shell egg surface microbial populations (aerobic bacteria, yeasts and molds, Enterobacteriaceae, Escherichia coli, and Salmonella) were monitored at 12 points along the processing line (accumulator, prewash rinse, washer 1, washer 2, sanitizer, dryer, oiler, scales, two packer head lanes, rewash entrance, and rewash exit). Three commercial facilities were each visited three times, a total of 990 eggs were sampled, and 5,220 microbiological samples were subsequently analyzed. Although variations existed in concentrations of microorganisms recovered from each plant, the patterns of fluctuation for each population were similar at each plant. On average, aerobes, yeasts and molds, Enterobacteriaceae, and E. coli prevalence were reduced by 30, 20, 50, and 30%, respectively, by the end of processing. The microbial concentrations (log CFU per milliliter) in the egg rinse collected from packer head lanes were decreased by 3.3, 1.3, 1.3, and 0.5, respectively, when compared with those of rinses collected from eggs at the accumulator. Salmonella was recovered from 0 to 48% of pooled samples in the three repetitions. Higher concentrations of Salmonella were recovered from preprocessed than from in-process or ready-to-pack eggs. These data indicate that current commercial practices decrease microbial contamination of egg shell surfaces.

Animals↗

[Microbiocenosis of parietal mucin in the gastrointestinal tract of rats].

The qualitative and quantitative composition of the microbial community in parietal mucin at different areas of the gastrointestinal tract (GIT) of rats was revealed. The pronounced variability in the quantitative and qualitative characteristics of microbiocenosis in parietal mucin of rats at different sections was revealed. The differences were most pronounced in the passage from upper to lower GIT sections, the large intestine found to be the richest biocenosis. The microbial composition of rat feces was faintly associated with the GIT parietal microbiocenosis. The individual areas of GIT mucosa were unique of their microbial characteristics and organization. This makes it possible to regard them as relatively independent biotopes and indicates that it is impossible to evaluate the microbial community by one of the colonic mucosal sifes.

Animals↗

Determinations of microbial loads associated with microscopic-size particles of Kennedy Space Center soil.

Plate counts for six fractions, of micrometer-size, of Kennedy Space Center soil provided estimates of aerobic, mesophilic, heterotrophic, microbial loads on single soil particles. Analyses included unheated particles, particles subjected to wet heat at 80 degrees C for 20 min, and particles subjected to dry heat at 110 degrees C for 1 hr. Unheated particles yielded mean counts ranging from 6 colonies per particle for the smallest (44-53 micrometers) soil fraction to approximately 55 colonies per particle for the largest size (105-125 micrometers) soil fraction tested. Mean counts for heat-resistant forms ranged from 2 colonies per particle for the smaller particles to 12-15 colonies for the largest particles analyzed.

Bacteria↗

The efficacy of a filtered handheld far-ultraviolet disinfection device for decontamination of high-touch surfaces in healthcare settings: a genomic bacterial analysis.

BACKGROUND AND OBJECTIVES: Enhanced environmental disinfection is linked to reduced hospital-acquired infection rates. In this study, we aimed to evaluate the efficacy of an emerging disinfection technology, a filtered far-UV-C handheld (FFUHH) device, for reducing bacterial loads on high-touch surfaces in shared clinical workrooms, and to isolate, identify and characterize clinically significant environmental pathogens. METHODS: We compared samples from high-touch items (dictation device, mouse, armchair, desk, and keyboard) before and after FFUHH treatment. Samples were collected weekly: contact plates for colony counts and swabs before and after intervention on standardized adjacent areas for each surface, respectively. The swabs were enriched and cultured on selective media to isolate pathogens. Environmental samples, as well as clinical samples collected from patients during the study period, were validated using MALDI-TOF and whole genome sequencing. RESULTS: Among the 440 collected plates (220 before and 220 after treatment), the highest mean colony count pre-treatment was detected from armchairs, and the lowest from keyboards. The mean reduction of colony-forming units ranged 53% and 83% and was statistically significant (P < 0.05) across all surfaces except for the keyboard. We characterized multidrug-resistant Staphylococcus epidermidis ST5 and ST16 strains, a carbapenem-resistant Acinetobacter baumannii, and a Klebsiella pneumoniae genetically related to a clinical isolate with a rare sequence type not previously detected in our institution. CONCLUSION: The FFUHH effectively reduced the microbial burden on high-touch surfaces. It can offer an advantage for surface disinfection and an alternative to routinely used biocides.

Humans↗