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PubMed2006

Stomach lysozymes of the three-toed sloth (Bradypus variegatus), an arboreal folivore from the Neotropics.

Lysozymes are antimicrobial defences that act as digestive enzymes when expressed in the stomach of herbivores with pre-gastric fermentation. We studied this enzyme in the complex stomach of the three-toed sloth (Bradypus variegatus), a folivore with pre-gastric fermentation. Lysozymes were identified by SDS-PAGE and immunoblotting in all portions: diverticulum, pouch, glandular and muscular prepyloric area with 14.3 kDa of molecular mass. Purified lysozymes from all areas but the diverticulum were characterized by MALDI-TOF, optimal pH, optimal ionic strength, and specific activity. The differences observed suggested at least three isoforms. The optimal pHs were similar to the pH of the stomach portion where the enzymes were isolated. The lysozyme from the pouch (fermentation chamber) exhibited higher specific activity and concentration than the others. The specific activity of the enzyme from the acid muscular prepyloric portion was comparable to that reported in the cow abomasums; however, its concentration was lower than that observed in cow. This distinctive pattern of secretion/specific activity and overall low concentration suggests different roles for the lysozymes in this herbivore compared to Artiodactyla. We postulate that sloth stomach lysozymes may still be antimicrobial defences by protecting the microbial flora of the fermentation chamber against foreign bacteria.

Animals↗

PubMed2005

Inactivation kinetics of Yersinia enterocolitica by citric and lactic acid at different temperatures.

Inactivation of Yersinia enterocolitica by citric (1--20% w/v) and lactic (0.3--4.0% v/v) acids at different temperatures (4, 20, 40 degrees C) has been investigated. Inactivation effect of citric and lactic acids was dependent on time and temperature of exposure and acid concentration. Survival curves of Y. enterocolitica suspended in citric acid solutions at 4 and 20 degrees C displayed a shoulder followed by an exponential inactivation, but at 40 degrees C a shoulder was not observed. At all temperatures investigated, survival curves of Y. enterocolitica suspended in lactic acid solutions were linear or slightly concave upwards. A mathematical model based on the Weibull distribution accurately described the kinetics of inactivation of Y. enterocolitica by both acids. The influence of the citric acid concentration on Y. enterocolitica resistance was independent of the treatment temperature. However for lactic acid, the influence of the acid concentration on microbial inactivation depended on the temperature. At any temperature investigated, lactic acid was significantly more effective than citric acid.

Citric Acid↗

PubMed2003

Growth of microorganisms in propofol and methohexital mixtures.

PURPOSE: The purpose of this study was to evaluate the growth of 4 different microorganisms in propofol, methohexital, and 1:1 and 1:3 mixtures of propofol and methohexital. MATERIALS AND METHODS: The microbial growth of Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and Escherichia coli in 1:1 and 3:1 mixtures of methohexital and 2 types of propofol was measured for 48 hours after mixing the solutions. Two of the 4 test solutions were composed of 1% methohexital combined with Diprivan (AstraZeneca Pharmaceuticals, Wilmington, DE), which uses EDTA as a microbial growth inhibitor. The growth of the same 4 microorganisms was also evaluated in 1:1 and 3:1 mixtures of 1% methohexital and a recently introduced generic propofol marketed by Baxter Pharmaceuticals (Deerfield, IL) that uses sodium metabisulfite as a preservative. RESULTS: Combining either Diprivan or generic propofol with methohexital in a 1:1 or 1:3 mixture ratio resulted in a solution that, like methohexital alone, significantly resisted the growth of C albicans, E coli, S aureus, and P aeruginosa for 48 hours. CONCLUSION: The results of our study suggest that 1:1 and 1:3 mixtures of propofol (either Diprivan or generic propofol) and methohexital behave similarly to methohexital alone by resisting growth of the microorganisms studied over the 48-hour time period. If, as has been reported elsewhere, the mixture of these 2 agents results in a solution with desirable anesthetic properties, its ability to resist microbial growth becomes clinically significant by extending the shelf-life of propofol in these mixtures.

Analysis of Variance↗

PubMed1998

Influence of temperature, pH and water activity on "in vitro" inhibition of Penicillium glabrum (Wehmer) Westling by yeasts.

Four different yeast species (Metschnikowia pulcherrima, Saccharomycopsis vini, Kluyveromyces marxianus, Cryptococcus albidus), isolated from surface of grapes, were evaluated for biocontrol potential against Penicillium glabrum. In order to investigate the influence of temperature, pH, water activity and yeast cell concentration on Penicillium glabrum inhibition, the individual effects and the interaction of these factors were analyzed by means of a Central Composite Design (CCD). All yeast species tested showed antagonistic effects which were more pronounced at high cell concentrations. The other variables affected the antagonistic effect differentially depending on the yeast species. Results of the experimental design showed that the selective success of a competitive microflora is under environmental control; moreover, when microbial cells are subjected to multiple factors, the effects and the reciprocal interactions of the individual variables cannot be independently evaluated.

Agar↗

PubMed2001

Studies on composition and stability of a large membered bacterial consortium degrading phenol.

A ten member microbial consortium (AS) consisting of eight phenol-degrading and two non-phenol-degrading strains of bacteria was developed and maintained in a fed-batch reactor by feeding 500 mg l(-1) phenol for four years at 28 +/- 3 degrees C. The consortium could degrade 99% of 500 mg l(-1) phenol after 24 hours incubation with a biomass increase of 2.6 x 10(7) to 4 x 10(12) CFU ml(-1). Characterization of the members revealed that it consisted of 4 principal genera, Bacillus, Pseudomonas, Rhodococcus, Streptomyces and an unidentified bacterium. Phenol degradation by the mixed culture and Bacillus subtilis, an isolate from the consortium was compared using a range of phenol concentrations (400 to 700 mg l(-1)) and by mixing with either 160 mg l(-1) glucose or 50 mg l(-1) of 2,4-dichlorophenol in the medium. Simultaneous utilization of unrelated mixed substrates (glucose/2,4-dichlorophenol) by the consortium and Bacillus subtilis, indicated the diauxic growth pattern of the organisms. A unique characteristic of the members of the consortia was their ability to oxidize chloro aromatic compounds via meta pathway and methyl aromatic compounds via ortho cleavage pathway. The ability of a large membered microbial consortia to maintain its stability with respect to its composition and effectiveness in phenol degradation indicated its suitability for bioremediation applications.

Aerobiosis↗

PubMed1994

In vitro antimicrobial activity of Shiva-11 against ocular pathogens.

Cecropins are antimicrobial peptides (30-35 amino acids) isolated from the hemolymph of the cecropia moth. Previous studies have demonstrated their antimicrobial efficacy against a variety of pathogens, including both gram-positive and -negative bacteria, fungi, protozoa, and enveloped viruses. To assess their therapeutic potential against ocular pathogens, we analyzed the in vitro antimicrobial activity of a synthetic cecropin analog, Shiva-11, against virulent microbial strains (Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumoniae, and Candida albicans). Bacterial isolates were obtained from human cases of severe ulcerative keratitis. Shiva-11 was tested at varying concentrations in bacterial suspensions containing 1-2 x 10(6) CFU/ml at 37 degrees C. Samples were plated on nutrient agar and colonies counted after 24-48 h of incubation. Shiva-11 yielded > 3 log killing of all isolates after 60 min of exposure to this compound. The results of this study indicate that Shiva-11 possesses broad-spectrum in vitro antimicrobial activity against human clinical ocular pathogens.

Animals↗

PubMed2006

Interference in adhesion of bacteria and yeasts isolated from explanted voice prostheses to silicone rubber by rhamnolipid biosurfactants.

AIMS: The effects and extent of adhesion of four different bacterial and two yeast strains isolated from explanted voice prostheses to silicone rubber with and without an adsorbed rhamnolipid biosurfactant layer obtained from Pseudomonasaeruginosa DS10-129 was studied. METHODS AND RESULTS: The ability of rhamnolipid biosurfactant to inhibit adhesion of micro-organisms to silicone rubber was investigated in a parallel-plate flow chamber. The anti-adhesive activity of the biosurfactant at different concentrations was significant against all the strains and depended on the micro-organism tested. The results showed an effective reduction in the initial deposition rates, and the number of bacterial cells adhering after 4 h, for all micro-organisms tested at the 4 g l(-1) undiluted rhamnolipid solution. Maximum initial reduction of adhesion rate (an average of 66%) occurred for Streptococcus salivarius GB 24/9 and Candida tropicalis GB 9/9. The number of cells adhering after 4 h on silicone rubber conditioned with biosurfactant was reduced to 48% for Staphylococcus epidermidis GB 9/6, Strep. salivarius GB 24/9, Staphylococcus aureus GB 2/1 and C. tropicalis GB 9/9 in comparison to controls. Perfusing the flow chamber with biosurfactant containing solution followed by the passage of a liquid-air interface, to investigate detachment of micro-organisms adhering to silicone rubber, produced high detachment (96%) of adhered cells for all micro-organisms studied, except for Staph. aureus GB 2/1 (67%). SIGNIFICANCE AND IMPACT OF THE STUDY: It is concluded that biosurfactant represent suitable compounds that should be considered in developing future strategies to prevent the microbial colonization of silicone rubber voice prostheses.

Actinomycetaceae↗

PubMed2002

Development and evaluation of a 16S ribosomal DNA array-based approach for describing complex microbial communities in ready-to-eat vegetable salads packed in a modified atmosphere.

There is a clear need for new approaches in the field of microbial community analyses, since the methods used can be severely biased. We have developed a DNA array-based method that targets 16S ribosomal DNA (rDNA), enabling the direct detection and quantification of microorganisms from complex communities without cultivation. The approach is based on the construction of specific probes from the 16S rDNA sequence data retrieved directly from the communities. The specificity of the assay is obtained through a combination of DNA array hybridization and enzymatic labeling of the constructed probes. Cultivation-dependent assays (enrichment and plating) and cultivation-independent assays (direct fluorescence microscopy and scanning electron microscopy) were used as reference methods in the development and evaluation of the method. The description of microbial communities in ready-to-eat vegetable salads in a modified atmosphere was used as the experimental model. Comparisons were made with respect to the effect of storage at different temperatures for up to 12 days and with respect to the geographic origin of the crisphead lettuce (Spanish or Norwegian), the main salad component. The conclusion drawn from the method comparison was that the DNA array-based method gave an accurate description of the microbial communities. Pseudomonas spp. dominated both of the salad batches, containing either Norwegian or Spanish lettuce, before storage and after storage at 4 degrees C. The Pseudomonas population also dominated the batch containing Norwegian lettuce after storage at 10 degrees C. On the contrary, Enterobacteriaceae and lactic acid bacteria dominated the microbial community of the batch containing Spanish lettuce after storage at 10 degrees C. In that batch, the Enterobacteriaceae also were abundant after storage at 4 degrees C as well as before storage. The practical implications of these results are that microbial communities in ready-to-eat vegetable salads can be diverse and that microbial composition is dependent both on the origin of the raw material and on the storage conditions.

Bacteria↗

PubMed1996

Microbiological response to mechanical treatment in combination with adjunctive therapy. A review of the literature.

The recognition of the microbial origin and the specificity of periodontal infections has resulted in the development of several adjunctive therapies (antibiotics and/or antiseptics) to scaling and root planing in the treatment of chronic adult periodontitis. This article aims to review the "additional" effect of a subgingival irrigation with chlorhexidine, or a local or systemic application of tetracycline or metronidazole, performed in combination with a single course of scaling and root planing in patients with chronic adult periodontitis. All treatment modalities are compared with scaling and root planing, based on their impact on: the probing depth (PD); total number of colony forming units per ml (CFU/ml); the proportions and/or the detection-frequency of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia; and/or on the percentages of cocci, spirochetes, motile, and other micro-organisms on dark field microscopy examination. All treatment modalities, including scaling and root planing without additional chemical therapy, resulted in significant reductions in the probing depth and the proportions of periodontopathogens, at least during the first 8 weeks post-therapy. However in comparison to a single course of scaling and root planing, the supplementary effect of adjunctive therapies seems to be limited. In general, only the irrigation with chlorhexidine 2%, the local application of minocycline, and the systemic use of metronidazole (in case of large proportions of spirochetes) or doxycycline (in case of large proportions of A. actinomycetemcomitans) seem to result in a prolonged supplementary effect when compared to scaling and root planing. Therefore, the use of antibiotics on a routine basis, especially in a systemic way, in the treatment of chronic adult periodontitis, can no longer be advocated, considering the increasing danger for the development of microbial resistance.

Administration, Oral↗

PubMed2001

Evaluation of combination treatment processes for the microbial decontamination of pork trim.

Combination treatment processes for the microbial decontamination of pork trim were developed and evaluated. Lean pork trim tissue (LPT) and fat-covered pork trim tissue (FPT) inoculated with swine feces were treated with intervention processes as follows: (i) control (untreated), (ii) water (15 degrees C, 120 s), (iii) water followed by lactic acid wash (15 degrees C, 75 s), (iv) combination 1 (water plus hot water [65.5 degrees C, 15 s] plus hot air [510 degrees C, 60 s] plus lactic acid), (v) combination 2 (water plus hot water [82.2 degrees C, 15 s] plus hot air [510 degrees C, 75 s] plus lactic acid), and (vi) combination 3 (water plus hot water [82.2 degrees C, 45 s] plus hot air [510 degrees C, 90 s] plus lactic acid). Populations of aerobic bacteria, psychrotrophic bacteria, coliforms, Escherichia coli, and lactic acid bacteria were determined before and after treatment and at days 2 and 7 of 4 degrees C storage. Regardless of the intervention treatment, lower microbial populations were observed on FPT than on LPT immediately after treatment and during the 7-day storage period. Both LPT and FPT treated with water plus lactic acid, combination 1, combination 2, and combination 3 had lower remaining populations of all microbial groups immediately after treatment than did water-treated samples. Populations of aerobic bacteria, coliforms, E. coli, and lactic acid bacteria on either LPT or FPT did not statistically increase during the 7-day storage period. On LPT, populations of psychrotrophic bacteria grew during 4 degrees C storage but remained lower at day 7 on LPT treated by combinations 2 and 3 (2.29 and 1.89 log10 CFU/cm2, respectively) than on LPT treated with water (4.07 log10 CFU/cm2) or water plus lactic acid (3.52 log10 CFU/cm2). Populations of psychrotrophic bacteria remained below detectable levels throughout the 7-day storage on FPT treated with water plus lactic acid or any of the three combination treatments. Treatment of pork trim with any of the combination treatments significantly (P < 0.05) affected the color and emulsion stability of the ground pork. Water and water plus lactic acid were the most favorable treatments in reducing microbial populations on pork trim without affecting the quality attributes of the ground pork.

Animals↗

PubMed2003

Implementation and analysis of an on-farm food safety program for the production of greenhouse vegetables.

Fresh fruits and vegetables are increasingly being recognized as vectors for foodborne illness. In 1998, an on-farm food safety program was developed for the Ontario Greenhouse Vegetable Growers (OGVG) in Ontario, Canada. This hazard analysis critical control point-based system was designed to reduce the potential of microbial contamination along the entire production and distribution process. A previous report (D. A. Powell, M. Bobadilla-Ruiz, A. Whitfield, M. W. Griffiths, and A. Luedtke, J. Food Prot. 65:918-923) outlined the first 2 years of the program; the present report documents the third year of the program's implementation. Through individual on-site grower visits and the microbiological testing of produce and water, the program's OGVG member requirements were determined, and the program's success was assessed. Growers' knowledge, understanding, and awareness of microbial risks associated with fresh produce have increased, and improvements in practices used in greenhouses and packing sheds were noted.

Agriculture↗

PubMedDate not supplied

[Stability of microbial association in the process of industrial biofiltration cleaning of gaseous discharges].

Results of industrial exploitation of a biofiltration plant tailored for purifying gaseous discharges of hazardous organic components such as toluene, cyclohexane, and xylene, are examined. Both numerical and compositional variations were monitored for a long-term (more than 1.5 years) utilization process in an association of microorganisms decomposing organic pollutants. A population of microbial association composed by one yeast and two bacterial strains in the biofilm on the surface of filtering sheets was abundant (10(8)-10(9) yeast cells/cm2 and 10(10)-10(11) bacterial cells/cm2) and stable during the whole period of monitoring. A microbial association in the culture medium averaging 10(6) yeast cells/l and 10(8) bacterial cells/l is more susceptible to technogenic impacts and seasonal fluctuations. Overall, the biofilter as an open and autonomic system maintained its microbial association, thereby providing a high-degree (93-98%) purification of industrial gaseous discharges from organic pollutants.

Colony Count, Microbial↗

PubMedDate not supplied

[Antagonism of microbial population of protective flora and its relation to microbiocenosis and nutritional factors].

Total antagonistic activity (TAA) of the populations of bifidobacteria, lactobacilli, enterobacteria was considered in relation to the method of its evaluation which has been modified, criteria of TAA assessment permitting its measurements in per cents. The technique proposed bases on determination of the metabolites capable of inhibiting pathogenic and opportunistic bacteria and provides information on the action of alimentary factors under study on the function of intestinal microbiocenosis. The factors may produce a multidirectional effect on the structure and biological activity of the microbial populations. The method can work to characterize microbiocenosis both in normal and pathological conditions, to evaluate efficacy of specially devised food products.

Child↗

PubMed2006

Microbial diversity in sediments of saline Qinghai Lake, China: linking geochemical controls to microbial ecology.

Saline lakes at high altitudes represent an important and extreme microbial ecosystem, yet little is known about microbial diversity in such environments. The objective of this study was to examine the change of microbial diversity from the bottom of the lake to sediments of 40 cm in depth in a core from Qinghai Lake. The lake is saline (12.5 g/L salinity) and alkaline (pH 9.4) and is located on the Qinghai-Tibetan Plateau at an altitude of 3196 m above sea level. Pore water chemistry of the core revealed low concentrations of sulfate and iron (<1 mM), but high concentrations of acetate (40-70 mM) and dissolved organic carbon (1596-5443 mg/L). Total organic carbon and total nitrogen contents in the sediments were approximately 2 and <0.5%, respectively. Acridine orange direct count data indicated that cell numbers decreased from 4 x 10(9) cells/g at the water-sediment interface to 6 x 10(7) cells/g wet sediment at the 40-cm depth. This change in biomass was positively correlated with acetate concentration in pore water. Phospholipid fatty acid (PLFA) community structure analyses determined decrease in the proportion of the Proteobacteria and increase in the Firmicutes with increased depth. Characterization of small subunit (SSU) rRNA genes amplified from the sediments indicated a shift in the bacterial community with depth. Whereas the alpha-, beta-, and gamma-Proteobacteria and the Cytophaga/Flavobacterium/Bacteroides (CFB) were dominant at the water-sediment interface, low G + C gram-positive bacteria (a subgroup of Firmicutes) became the predominant group in the anoxic sediments. Both PLFA and the sequence data showed similar trend. The Proteobacteria, CFB, and gram-positive bacteria are present in other saline lakes, but the presence of Actinobacteria and Acidobacteria/Holophaga in significant proportions in the Qinghai Lake sediments appears to be unique. The archaeal diversity was much lower, and clone sequences could be grouped in the Euryarchaeota and Crenarchaeota domains. The archaeal clones were not related to any known cultures but to sequences previously found in methane-rich sediments. Acetate-utilizing methanogens were isolated from sediment incubations, and alpha- and gamma-proteobacterial isolates were obtained from a water sample from the lake-bottom (23 m). Our data collectively showed that the observed diversity and shift in the community structure with depth was correlated with geochemical parameters (the redox state and availability of electron acceptor and donor). Heterotrophic methanogenesis is possibly adominant metabolic process in the Qinghai Lake sediments. These results reinforce the importance of geochemical controls on microbial ecology in saline and alkaline lake environments.

Acridine Orange↗

PubMedDate not supplied

Microbiological screening for cariogenic bacteria in children 9 to 36 months of age.

PURPOSE: The objective of this study was to evaluate sampling methods for recovery of mutans streptococci (MS) and lactobacilli (LB) in children 9 to 36 months of age. METHODS: Tongue and plaque specimens collected on cotton swabs and stimulated saliva were diluted and plated on selective and nonselective media. Tongue specimens on a swab and mouth mirror were inoculated directly on selective agar media (MS only). Sampling methods were compared by frequency of recovery of MS or LB, correlation of microbial counts with dmfs scores, and potential of specific microbial counts to predict caries presence or absence. RESULTS: The mean dmfs score of 87 subjects was 6.3; 48 subjects were caries free. Levels of MS and LB were consistently higher in plaque than in other sampling techniques (P<.001), and frequencies of recovery of MS were highest in plaque (P<.041) and tongue (P<.006). Frequency of LB recovery did not differ significantly between sampling methods. Counts of MS or LB in total subjects and subjects aged 9 to 24 months correlated positively with dmfs scores (P<.028). Threshold levels of MS which were predictive of presence of caries were: (1) plaque=>2x10(5); (2) tongue=>10(4); (3) saliva=>10(5); (4) mirror=>50; and (5) swab=>50. Comparable levels of LB were: plaque, >10(3); tongue, >10(2) and saliva, >10(3). Specificities associated with these predictions were higher than sensitivities for all sampling methods. CONCLUSIONS: (1) All sampling methods were adequate for microbial risk assessment tests in children under 3 years of age; (2) MS was a stronger indicator of caries status than LB.

Age Factors↗

PubMed1995

Is the sutureless cataract incision a valve for bacterial inoculation?

Many cataract surgeons perform sutureless surgery to decrease operating time, postoperative astigmatism, and healing time. Anecdotal case reports of postoperative endophthalmitis after sutureless surgery prompted our investigation of this type of wound closure and its possible relationship to an increased incidence of infection. This in vitro study addressed the question: Is sutureless more likely than sutured cataract surgery to provide a route for inoculation of microbial organisms into the eye? Twenty-eight human eyes obtained postmortem were randomly divided into 14 pairs and successively incubated for 90, 150, 210, and 270 minutes each in a suspension of Staphylococcus epidermidis in physiologic media. Cultured aqueous aspirates yielded no significant differences between sutured and unsutured eyes in colony counts at any time interval. This suggests that both sutured and unsutured wounds resist bacterial ingrowth equally and that a properly constructed unsutured wound is not a significant valve for bacterial inoculation in an eye pressurized to physiological conditions.

Adult↗

PubMed1996

Evaluation of Petrifilm methods for enumeration of aerobic flora and coliforms in a wide range of foods.

Petrifilm is a ready-to-use alternative to traditional microbial enumeration methods. The Petrifilm Aerobic Count Plate (ACP) and Coliform Count Plate (CCP) were compared with standard methods for the enumeration of the aerobic mesophilic flora and coliform bacteria in 91 foods covering a wide range of different food commodities. There was good correlation between the Petrifilm ACP and the standard aerobic colony count method (r = 0.989) and between the Petrifilm CCP and the standard Violet Red Bile Agar plating method (r = 0.872). In both cases, the Petrifilm methods had a better repeatability than the standard methods. The Petrifilm ACP and CCP were shown to be practical and accurate alternatives to standard enumeration methods in a wide range of foods, with benefits of saving time, labour and incubator space.

Bacteria, Aerobic↗

PubMed2002

Preservation of fresh meat with active and modified atmosphere packaging conditions.

The sensory, microbiological and physicochemical attributes of fresh meat stored at 5 and 15 degrees C were affected by the combined effect of volatile compounds of oregano essential oil and modified atmosphere packaging conditions (40% CO2/30% N2/30% O2, 100% CO2, 80% CO2/20% air, vacuum pack and air). It was found that the extension of shelf life of meat samples depended on the packaging conditions and augmented in the order: air < vacuum pack < 40% CO2/30% N2/30% O2 < 80% CO2/ 20% air < 100% CO2. Longer shelf life was observed in samples supplemented with the volatile compounds of oregano essential oil and stored under the same packaging conditions mentioned above. The extension of shelf life may be due to the synergistic effect of volatile compounds of oregano essential oil and the modified atmosphere packaging used on the microbiological and physicochemical characteristics of meat. Indeed, both these hurdles can prolong and delay microbial growth or suppress the final counts of the spoilage microorganisms in comparison with the 'control' samples. The effect of essential oil volatile compounds was even more pronounced on the physicochemical changes of meat samples caused by microbial association. Oregano essential oil delayed glucose and lactate consumption, both indicators of meat spoilage aerobically as well as under 40% CO2/30% N2/30% O2, and 100% CO2. Finally, changes in other metabolites such as formic acid were also observed.

Aerobiosis↗