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Effects of mode of delivery and necrotising enterocolitis on the intestinal microflora in preterm infants.

To investigate the effects of mode of delivery and of necrotising enterocolitis on the faecal microflora, 140 infants born before 33 weeks of gestation were followed up for symptoms of necrotising enterocolitis. Stool samples for gas-liquid chromatography and culture were collected twice weekly, and, when necrotising enterocolitis was suspected, for 2 months. For each infant with necrotising enterocolitis (n=21), two control infants matched for birth weight and gestational age were selected from the remaining study population. In gas-liquid chromatography analysis, the faecal bacterial microflora of infants born via caesarean section differed significantly from the gut microflora of those born via the vaginal route. The intestinal microflora showed a significant alteration in the necrotising enterocolitis group at time of diagnosis. At the onset of necrotising enterocolitis, faecal colonisation with Enterococcus species and Candida albicans was significantly more frequent in symptomatic infants than in controls. In infants with positive blood cultures and positive intestinal biopsy cultures, concomitant stool samples revealed the same microbial pathogens. In conclusion, the intestinal microbial colonisation in preterm infants born by caesarean section differs from that in preterm infants born via the vaginal route. A significant change in faecal microbial colonisation seems to occur at the onset of necrotising enterocolitis. Pathogens detected in the stools at that time might have a causative role in the development of the disease.

Analysis of Variance↗

Inactivation of Salmonella Senftenberg 775W by ultrasonic waves under pressure at different water activities.

The inactivation of Salmonella Senftenberg 775W by ultrasonic waves (20 kHz, 117 microm) under pressure (175 kPa) treatments at sublethal (manosonication; MS) and lethal temperatures (manothermosonication; MTS) in media of different water activities has been investigated. Heat decimal reduction time values increased up to eighteen fold when the water activity was decreased from >0.99 to 0.93 at 65 degrees C, but hardly increased the MS resistance. In reduced water activity media (a(w) of 0.96 and 0.93) a synergistic lethal effect was observed between heat and ultrasound under pressure, being the inactivation rate of Salmonella Senftenberg 775W three times faster than the expected additive rate considering an effect of both bacterial lethal processes. An empirical mathematical equation enabled to predict the D(MS) and D(MTS) values obtained at different temperatures and a(w) in the ranges investigated of Salmonella serovars and also the microbial level of inactivation due to the synergistic lethal effect of MTS treatments in media of reduced a(w). This work could be useful for improving sanitation and preservation treatments of foods, especially those in which components protect microorganisms to heat.

Colony Count, Microbial↗

Accelerated transformation and deactivation of erythromycin in superheated water. 2. Transformation reactions and bioassays.

The work presented in this second part of a two-part series on the accelerated transformation of erythromycin in superheated water focuses on the chemical nature and resultant antimicrobial implications of the overall reaction observed. Analyses of reactor effluents employing infrared spectroscopy and liquid chromatography/mass spectrometry indicated that the initial step in the decomposition pathway is one of dehydration. Subsequent hydrolysis in the presence of an acetate buffer appeared to result in the loss of the sugar cladinose. Chemical transformation under superheated conditions was tied to the loss of antibiotic function by an agar dilution test. The sensitivity of this test was verified by determination of the minimum inhibitory concentration (MIC) of erythromycin corresponding to each of two test microorganisms. MIC values for the selected strains of Escherichia coli and Bacillus subtilis were 35 and 0.5 mg/L, respectively. To relate the loss of antimicrobial activity to increased reaction temperature and thus to increased extent of parent compound transformation to microbially benign intermediates, bioassays using E. coli as test microorganism were performed on flow reactor effluents resulting from reaction of initial erythromycin concentrations of 75-150 mg/L.

Amino Sugars↗

Colonization of personal digital assistants carried by healthcare professionals.

This study evaluated personal digital assistant (PDA) microbial colonization before and after cleaning with alcohol. Samples from 75 PDAs were processed. Before cleaning, 96% of the samples were culture positive. After cleaning, 75% became culture negative. PDAs cleaned with an alcohol swab demonstrated significant reduction in colonization.

Bacteria↗

Protozoan bacterivory and Escherichia coli survival in drinking water distribution systems.

The development of bacterial communities in drinking water distribution systems leads to a food chain which supports the growth of macroorganisms incompatible with water quality requirements and esthetics. Nevertheless, very few studies have examined the microbial communities in drinking water distribution systems and their trophic relationships. This study was done to quantify the microbial communities (especially bacteria and protozoa) and obtain direct and indirect proof of protozoan feeding on bacteria in two distribution networks, one of GAC water (i.e., water filtered on granular activated carbon) and the other of nanofiltered water. The nanofiltered water-supplied network contained no organisms larger than bacteria, either in the water phase (on average, 5 x 10(7) bacterial cells liter-1) or in the biofilm (on average, 7 x 10(6) bacterial cells cm-2). No protozoa were detected in the whole nanofiltered water-supplied network (water plus biofilm). In contrast, the GAC water-supplied network contained bacteria (on average, 3 x 10(8) cells liter-1 in water and 4 x 10(7) cells cm-2 in biofilm) and protozoa (on average, 10(5) cells liter-1 in water and 10(3) cells cm-2 in biofilm). The water contained mostly flagellates (93%), ciliates (1.8%), thecamoebae (1.6%), and naked amoebae (1.1%). The biofilm had only ciliates (52%) and thecamoebae (48%). Only the ciliates at the solid-liquid interface of the GAC water-supplied network had a measurable grazing activity in laboratory test (estimated at 2 bacteria per ciliate per h). Protozoan ingestion of bacteria was indirectly shown by adding Escherichia coli to the experimental distribution systems. Unexpectedly, E. coli was lost from the GAC water-supplied network more rapidly than from the nanofiltered water-supplied network, perhaps because of the grazing activity of protozoa in GAC water but not in nanofiltered water. Thus, the GAC water-supplied network contained a functional ecosystem with well-established and structured microbial communities, while the nanofiltered water-supplied system did not. The presence of protozoa in drinking water distribution systems must not be neglected because these populations may regulate the autochthonous and allochthonous bacterial populations.

Animals↗

Efficacy of acidic electrolyzed water for microbial decontamination of cucumbers and strawberries.

An examination was made of the efficacy of acidic electrolyzed water (AcEW, 30 ppm free available chlorine), ozonated water (5 ppm ozone), and a sodium hypochlorite solution (NaOCl, 150 ppm free available chlorine) for use as potential sanitizers of cucumbers and strawberries. AcEW and NaOCl reduced the aerobic mesophiles naturally present on cucumbers within 10 min by 1.4 and 1.2 log CFU per cucumber, respectively. The reduction by ozonated water (0.7 log CFU per cucumber) was significantly less than that of AcEW or NaOCl (P < or = 0.05). Cucumbers washed in alkaline electrolyzed water for 5 min and then treated with AcEW for 5 min showed a reduction in aerobic mesophiles that was at least 2 log CFU per cucumber greater than that of other treatments (P < or = 0.05). This treatment was also effective in reducing levels of coliform bacteria and fungi associated with cucumbers. All treatments offered greater microbial reduction on the cucumber surface than in the cucumber homogenate. Aerobic mesophiles associated with strawberries were reduced by less than 1 log CFU per strawberry after each treatment. Coliform bacteria and fungi associated with strawberries were reduced by 1.0 to 1.5 log CFU per strawberry after each treatment. Microbial reduction was approximately 0.5 log CFU per strawberry greater on the strawberry surface than in the strawberry homogenate. However, neither treatment was able to completely inactivate or remove the microorganisms from the surface of the cucumber or strawberry.

Bacteria↗

Microbial exposure, symptoms and inflammatory mediators in nasal lavage fluid of kitchen and clerical personnel in schools.

OBJECTIVES: The aim of this study was to investigate how the microbial conditions of kitchen facilities differ from those in other school facilities. The health status of the personnel was also studied. MATERIALS AND METHODS: The microbial investigations were conducted in six moisture-damaged schools and two reference schools. The symptoms of the kitchen personnel were surveyed with questionnaires and inflammatory responses in nasal lavage (NAL) fluid were measured. RESULTS: The total concentrations of airborne microbes were lower in kitchens than in other facilities of the schools. However, the occurrence of moisture damage increased the airborne microbial concentrations both in kitchens, and in other facilities. Bacterial concentrations were high on surfaces in the damaged kitchens. Gram-negative bacteria predominated, but also thermophilic bacteria and mycobacteria were detected. Respiratory and general symptoms were prevalent both among kitchen workers and clerical personnel in the moisture-damaged environments. Reported allergies and repeated respiratory infections were connected with high IL-4 concentrations in NAL fluid. Median concentrations of studied inflammatory mediators (NO, IL-4, IL-6 and TNF-alpha) were slightly higher in NAL samples of kitchen workers than among the clerical personnel. CONCLUSIONS: Kitchen facilites differ from other facilities of the school building for their moisture conditions and microbial contamination. Thus, they represent a specific type of environment that may affect the health status of the personnel.

Air Pollutants, Occupational↗

A randomized clinical trial of the effect of yoghurt on the human salivary microflora.

Yoghurt is active against some human pathogens, so this experiment tested whether it is also active against the salivary microflora. A clinical trial was designed, with volunteers aged between 23 and 37 years. Initially, for 8 weeks, they consumed neither yoghurt nor casein-free soybean ice cream (phase 1). They were then split randomly into test (yoghurt) and control (ice cream) groups and required to consume 125 g of these foods twice daily for 8 weeks (phase 2) and then to avoid them for the following 2 weeks (phase 3). Many potential sources of microbial fluctuation were standardized. Salivary samples were then collected at regular intervals of 2 weeks. For each group and phase, the mean logarithms of the salivary counts for total viable flora, oral streptococci, mutans streptococci, lactobacilli and Candida were calculated. The prevalence of Streptococcus thermophilus and Lactobacillus bulgaricus was also assessed. Twenty test and 22 control participants completed the study. At phase 2, the mean for mutans streptococci was lower in the test than in the control group (3.6 vs. 4.0 log colony-forming units/ml; P=0.02). Moreover, the mean had decreased in all test participants with high phase 1 means for lactobacilli as compared to only 36.4% of those with low phase 1 means (P=0.01). L. bulgaricus was transiently detected in three test participants during phase 2. These results suggest that yoghurt does have some activity against the salivary microflora, but this does not seem to be due to the installation of yoghurt microorganisms in the mouth.

Adult↗

The effect of rings on microbial load of health care workers' hands.

BACKGROUND: The hands of health care workers (HCWs) serve as a major route for transmission of nosocomial infection. Although handwashing is known to reduce cross-transmission of infection, the influence of rings on the efficacy of handwashing and the carriage of bacteria on the hands has not been well established. METHODS: In this study, 50 HCWs with rings were paired by unit with 50 HCWs without rings. Cultures were obtained by use of a timed-friction rinse before and after a timed handwashing. Standard laboratory procedures were followed for identification of the bacteria. RESULT: When colony counts before handwashing are taken into consideration, a significant difference is seen after handwashing between the two groups (R2 = 0.56). The regression model showed that the slope was significantly steeper (p < 0.0014) for the group with rings. This effect is more apparent when the colony count on hands is greater than 1000 colony forming units before handwashing. CONCLUSIONS: A standardized, timed handwashing procedure was effective in decreasing the bioload of HCWs' hands. The effect of rings on the bioload was significant in this study.

Case-Control Studies↗

Some remarks on the bias of the MPN method.

The estimation of cell number by using the MPN method was studied mathematically for three parallel inoculations from three decimal dilutions. As a result of computation it can be established that these estimations at a range of microbial numbers over 10 are biased. Confidential estimation could be obtained at cell numbers between 0 and 10 accepting only a few combinations of positive and negative test tubes. These are 1 0 0, 2 0 0, 3 0 0, 3 1 0 and 3 2 0.

Bias↗

Microbiological composition of whole saliva and caries experience in minority populations.

Microbiological factors in whole saliva, previously implicated in dental caries formation, were studied in four caries-positive adult ethnic groups: Asians, blacks, whites, and Hispanics. Whole saliva was collected from each patient and used for the enumeration of total cultivable microflora (T), streptococci, mutans streptococci (MS), and lactobacilli. In the female subgroup, statistically significant differences were found among the ethnic groups with regard to T in saliva. In the male subgroup, statistically significant differences were found among the ethnic groups with regard to MS in saliva. Statistically significant differences were also found among the ethnic groups in relation to caries score (DMFS), with the Hispanic group having the highest mean DMFS score; and among socioeconomic groups with regard to MS in saliva. Moreover, microbial composition (T and MS) of whole saliva and DMFS scores differed significantly (P < 0.05) among the ethnic groups studied.

Adolescent↗

A rapid solid-phase fluorimetric assay for measuring bacterial adherence, using DNA-binding stains.

In this report, we describe the validation of a rapid, single-step, microtiter plate method for quantifying bacterial adherence, based on fluorescent labeling of microorganisms with cell-permeable fluorescent DNA-binding probes. We have tested the binding to saliva-coated microtiter plates of bacteria, including Helicobacter pylori and viridans streptococci (S. mitis, S. gordonii, S. sanguis), known to interact with salivary components. Furthermore, we tested the short-term and longer-term temporal stability of a saliva-mediated adherence of these bacteria in a healthy population (N=30). The assay exhibited excellent reliability statistics, yielding within-assay variability coefficients ranging from 4.9% to 11%. A range of approximately 5 x 10(4)-1 x 10(7) cells could be detected. This method may be generally applicable to study surface binding of virtually any microbial species, while obviating the need of radioactive materials or specific antibodies for quantification, thus providing a procedure that is useful to both basic and clinical research.

Bacterial Adhesion↗

Phylogenetic and functional biomakers as indicators of bacterial community responses to mixed-waste contamination.

Few studies have demonstrated changes in community structure along a contaminant plume in terms of phylogenetic, functional, and geochemical changes, and such studies are essential to understand how a microbial ecosystem responds to perturbations. Clonal libraries of multiple genes (SSU rDNA, nirK, nirS, amoA, pmoA, and dsrAB) were analyzed from groundwater samples (n = 6) that varied in contaminant levels, and 107 geochemical parameters were measured. Principal components analyses (PCA) were used to compare the relationships among the sites with respect to the biomarker (n = 785 for all sequences) distributions and the geochemical variables. A major portion of the geochemical variance measured among the samples could be accounted for by tetrachloroethene, 99Tc, No3, SO4, Al, and Th. The PCA based on the distribution of unique biomarkers resulted in different groupings compared to the geochemical analysis, but when the SSU rRNA gene libraries were directly compared (deltaC(xy) values) the sites were clustered in a similar fashion compared to geochemical measures. The PCA based upon functional gene distributions each predicted different relationships among the sites, and comparisons of Euclidean distances based upon diversity indices for all functional genes (n = 432) grouped the sites by extreme or intermediate contaminant levels. The data suggested that the sites with low and high perturbations were functionally more similar than sites with intermediate conditions, and perhaps captured the overall community structure better than a single phylogenetic biomarker. Moreover, even though the background site was phylogenetically and geochemically distinct from the acidic sites, the extreme conditions of the acidic samples might be more analogous to the limiting nutrient conditions of the background site. An understanding of microbial community-level responses within an ecological framework would provide better insight for restoration strategies at contaminated field sites.

Bacteria↗

Mutans streptococci in subgingival plaque of treated and untreated patients with periodontitis.

BACKGROUND: The etiology of root caries is thought to be identical to coronal caries, though root caries seem to be more complicated because of the higher susceptibility of exposed roots (dentin) by periodontal therapy to demineralization than intact enamel. This implies that mutans streptococci are the most likely pathogens in the development of root caries. Although it is known that both the numbers of mutans streptococci and the frequency of isolation in root caries lesions are negatively correlated with the distance from the gingival margin, the subgingival sulcus has not been considered a possible habitat for mutans streptococci. However, subgingival occurrence of mutans streptococci in both untreated and treated periodontal patients has not been documented well in the literature. OBJECTIVE: To investigate the presence and levels of mutans streptococci in the subgingival plaque of patients (n=154) in different stages of periodontal therapy. The subgingival sulcus may be a possible habitat for mutans streptococci. This localisation of mutans streptococci may be of importance in the development of root caries after periodontal surgery. MATERIALS AND METHODS: In this cross-sectional study, subgingival plaque samples from 154 consecutive adult periodontitis patients were tested for presence and levels of mutans streptococci and putative periodontal pathogens by anaerobic cultures. These patients were divided into 4 groups based on their stage of periodontal treatment: (1) untreated patients; (2) patients after initial periodontal therapy only; (3) patients in the maintenance phase who not underwent periodontal surgery; (4) patients after periodontal surgery. RESULTS: The prevalence of mutans streptococci in the 4 study groups varied from 82% in untreated patients to 94% in maintenance patients. The mean proportion of mutans streptococci was 6.65% in maintenance patients versus 1.86% in untreated patients (p=0.005) and 2.51% in patients after scaling and root planing (p=0.041). CONCLUSIONS: The subgingival area is a microbial habitat for mutans streptococci that may be of importance in the development of root caries in periodontitis patients.

Adult↗

Control of microbial contamination during surgical harvest of pig renal xenografts.

The pig has been identified as the most likely source of xenograft material for clinical use and studies are ongoing to overcome the immunological hurdles of pig-to-human transplantation. Attention is now being focussed on identifying and reducing the potential microbiological hazards associated with this technique. Studies have primarily addressed issues surrounding the production and health monitoring of xenograft source pigs and none have so far specifically evaluated the possible risks of microbial contamination during xenograft harvest. In this report, we evaluate the possible routes for contamination of a pig kidney xenograft during organ harvest and describe approaches to the control of these hazards, including the novel use of a custom designed airtight surgical canopy. A standard procedure for microbiological monitoring during xenograft harvest was devised and evaluated. This allowed the rapid identification and anti-microbial sensitivity testing of any isolated organisms. This would enable an early and appropriate pre-emptive treatment of infection because of transmission of pig micro-organisms.

Animals↗

Bacterial colonization of barrier material and periodontal regeneration.

The objective of this study was to evaluate the relationship between the presence of bacteria on the tooth-facing surface of ePTFE barriers and the clinical outcome of membrane supported reconstructive periodontal surgery. 20 systemically healthy subjects affected by chronic periodontitis were enrolled. One tooth site per patient, associated with an angular bony defect and a probing attachment loss of > 4 mm, was selected to be treated by means of a guided tissue regeneration procedure using an ePTFE barrier membrane. Antibiotics (Augmentin 1 g/day) for 2 weeks were prescribed. In addition to the use of chlorhexidine for post-surgical plaque control, all patients were recalled once a week for professional tooth cleaning. The barrier material was harvested for SEM analysis after 4-6 weeks. Professional tooth cleaning and reinforcement of sel-performed oral hygiene measures were given at 1 mouth intervals after membrane removal. For each treated site, the difference in probing attachment loss between baseline examination and a follow-up examination after 6 months of healing was calculated. The results of the SEM-analysis revealed that bacterial colonization was evident in the collar area of all the retrieved membranes. In the mid part of the membranes 30 out of 60 microscopic fields (50%) demonstrated microbial colonization, and in the most apical part 9 out of 60 fields (15%). Regression analysis indicated that gain in probing attachment level was positively correlated to initial attachement loss and negatively correlated to microbial colonization of the mid part of the membranes. It was concluded that bacterial colonization in the mid part of the ePTFE membrane reduced the potential gain in probing attachment following GTR-therapy with almost 50%.

Adult↗

A direct role for dual oxidase in Drosophila gut immunity.

Because the mucosal epithelia are in constant contact with large numbers of microorganisms, these surfaces must be armed with efficient microbial control systems. Here, we show that the Drosophila nicotinamide adenine dinucleotide phosphate (NADPH) oxidase enzyme, dual oxidase (dDuox), is indispensable for gut antimicrobial activities. Adult flies in which dDuox expression is silenced showed a marked increase in mortality rate even after a minor infection through ingestion of microbe-contaminated food. This could be restored by the specific reintroduction of dDuox, demonstrating that this oxidase generates a unique epithelial oxidative burst that limits microbial proliferation in the gut. Thus, oxidant-mediated antimicrobial responses are not restricted to the phagocytes, but rather are used more broadly, including in mucosal barrier epithelia.

Animals↗

Effects of an abrupt diet change from hay to concentrate on microbial numbers and physical environment in the cecum of the pony.

Microbial numbers, pH, fluid volume, and turnover rate in the pony cecum were measured during an abrupt change from an all-forage to an all-concentrate diet, both fed at maintenance energy levels. Concentrate feeding resulted in increased (P less than 0.01) numbers of total viable anaerobic bacteria. The numbers of organisms growing on selective starch medium increased (P less than 0.01) when concentrate was fed, while numbers on xylan and pectin media decreased (P less than 0.025). Seven days after the diet change to concentrate, the number of bacteria growing on lactate medium increased (P less than 0.01), followed by a gradual decline. Cellulolytic bacteria occurred in low numbers, ranging from 1.1 x 10(4) to 4.4 x 10(4) per g of cecal contents. Feeding all concentrate decreased both the number of genera (P less than 0.01) and total protozoan numbers (P less than 0.01) in the cecum. Minimum cecal pH values of 6.4 and 5.8 were obtained when forage and concentrate, respectively, were fed, with the minimum pH occurring 6 h postfeeding. Dry-matter percentage of cecal contents followed a diurnal pattern which was the inverse of the pH curve. During forage feeding, the cecum contained an average of 2.2 liters (1.6 to 3.4 liters), which turned over 3.9 times per day. When concentrate was fed, cecal volume averaged 3.9 liters (0.6 to 8.6 liters), with a mean liquid turnover of 4.2 times per day. Microbial numbers and pH changes in the pony cecum associated with an abrupt change in diet from hay to concentrate resembled those which occur in the rumen under similar feeding conditions.

Animal Feed↗