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Hydrogen sulphide production tests and the detection of groundwater faecal contamination by septic seepage.

The 'H2S test' is being advanced for microbiological water quality testing where conventional coliform based methods are impractical or too expensive. It involves ambient temperature incubation of water samples with nutrient formulated to generate hydrogen sulphide when 'faecal' bacteria are present. Recently a WHO review identified several concerns including the limited number of comparative studies, formulation variability, and false positives and negatives. In response we have compared the H2S test's ability to detect and quantify faecal contamination in an aquifer impacted by septic tank leachfields with measurements obtained concurrently using conventional bacterial indicators, coliphages, faecal sterol biomarkers, Cryptosporidium and Giardia. Like these other analytes, H2S testing detected a contamination gradient ranging from high (septic liquid) to moderate (exfiltration zones), to background (e.g. domestic bores), corresponding to indicator removal + dilution by factors > 10(6). Presence/absence tests could not distinguish between heavily and slightly contaminated waters, whereas multi-tube testing (e.g. 10 x 10 mL arrays) did. It was concluded that while the WHO review concerns are justified, the H2S test performance shows promise in sanitary survey work, can be improved by employing an mpn approach and has potential to aid in the protection of source water and identifying contaminated groundwater.

Environmental Monitoring↗

Fecal peritonitis: microbial adherence to serosal mesothelium and resistance to peritoneal lavage.

Fecal contamination of the peritoneal cavity is a serious and potentially life-threatening event. While numerous models have been developed to study the pathogenesis of intraabdominal infection, to date, most investigations have failed to focus on the adherence of the contaminants to the serosal mesothelium. In the present investigation, the cecal ligation and puncture technique (CLP) was performed in Sprague-Dawley rats to study the following: (a) the kinetics of microbial adherence to the serosal mesothelium, (b) the stability of the aerobic and anaerobic intraperitoneal/mesothelial populations, following extended saline lavage, and (c) the impact of antimicrobial lavage on the stability of the mesothelial microbial populations. The Enterobacteriaceae rapidly colonized the serosal mesothelium and were the predominant flora up to 4 hours post-CLP. After 8 hours, the Bacteroides fragilis group represented the predominant peritoneal wash and mesothelial-associated microorganisms. Extended saline lavage failed to significantly reduce the mesothelial microbial populations. While antimicrobial lavage produced an immediate decrease in mesothelial microbial recovery, the results were transitory and the microbial populations achieved or exceeded prelavage levels at 24 hours postlavage. Microbial colonization of the peritoneal mesothelial surface is a rapid and stable phenomena following penetrating injury to the distal bowel. The results further suggest that the mesothelial populations are resistant to intraperitoneal lavage.

Animals↗

Enterobacteriaceae suppression by three different oral doses of polymyxin E in human volunteers.

Polymyxin E is frequently used as an oral drug for flora suppression of the gastrointestinal canal. The suppression effect is dose dependent because polymyxin E is moderately inactivated by faecal and food compounds. Three oral polymyxin E doses (150, 300, 600 mg daily) were given to six volunteers for 6 days. The Enterobacteriaceae suppression effect was compared by means of the suppression index i.e. ratio of total number of faecal samples free of Enterobacteriaceae to the total number of faecal samples. The impact on the indigenous (mostly anaerobic) flora was measured in four ways: (i) beta-aspartylglycine content; (ii) volatile fatty acid pattern; (iii) yeast overgrowth and (iv) Streptococcus faecalis decrease. Enterobacteriaceae suppression was most successful during 600 mg oral polymyxin E (suppression indices during 150, 300 and 600 mg were 0.32, 0.55 and 0.89 respectively). None of the four markers of indigenous flora alterations were positive. However, using this dosage half of the volunteers suffered rather severe gastrointestinal side-effects. Oral polymyxin E in a dosage of minimum 600 mg daily seems to possess the ideal properties of a flora suppression agent, if the gastrointestinal side-effects could be mitigated.

Administration, Oral↗

[Antibiotic associated diarrhea and Clostridium difficile associated diarrhea in the elderly].

Antibiotic associated diarrhea (AAD) is a common complication when antibiotics are used and is frequent in the elderly. It has an impact on the length of hospital stay and increases the comorbidity. Together with the type of antibiotic that is given, the length of antibiotical treatment and the combination of antibiotics is more predictive for the evolution of diarrhea when compared to the total given dose. Mostly AAD is benign, but an infection with C. difficile should always be excluded. C. difficile-enterocolitis is frequent among residents in nursing homes and in hospitalised patients. The clinical presentation varies from asymptomatic colonisation tot severe debilitating disease. A rapid diagnosis can be performed by detection of C. difficile toxin by an enzyme-linked immunoassay. Oral metronidazole and oral vancomycine are equally effective in the therapy. In relapsing infection an extended tapering regimen is sometimes necessary.

Aged↗

Suspended particle effects on ClO2/ultraviolet light combined disinfection of effluent.

The concentration of suspended solids of effluent often varies in a wide range, therefore the dose of ultraviolet light (UV) in disinfection process needs to be adjusted to meet the disinfection criterion at a high frequency, and the desired disinfection effect is difficult to be ensured. The particles size and particle-associated fecal coliform (F.C.) contribution, and their influence on UV disinfection were investigated when ClO2 and UV combined disinfection process was used. The results showed that suspended solids content had a major impact on UV disinfection efficiency, especially the large particle size fraction. Particles (D>10 microm) associated F. C. were difficult to be disinfected and were the main part of the tailings of F.C. inactivation curve. Pre-ClO2 oxidation could reduce the number of particles in effluent, and make large particles decrease to small ones. Therefore, the influence of particles on UV disinfection could be reduced after pre-ClO2 oxidation, and the resistance ability to particle loadings of combined process was enhanced. Moreover, the combined process has a lot of advantages, such as low toxicity, low operational/maintenance costs; it is also convenient to be established in the existing wastewater plant or the new planned one.

Disinfection↗

Seasonal changes of gastrointestinal nematode burden in sheep under an intensive grazing system.

A study was conducted of the monthly fluctuations of worm burdens and arrested development of gastrointestinal (GI) nematodes in sheep from a commercial farm of the Ebro valley (Spain). Twenty-four previously helminth-free female lambs (permanent lambs) grazed together with a flock of 500 resident ewes for 12 months following a three lambing/2-year reproductive management system. Two helminth-free lambs (tracer lambs) were added to the flock each month and allowed to graze for 4 weeks. Measurements were made of the population of infective larvae on the pasture, and of levels of serum pepsinogen and faecal worm eggs in ewes and lambs. Additionally, post-mortem worm counts from two tracer and two permanent lambs were used to establish the level of infection at monthly intervals. Three generations of parasites were identified in the experiment. The generation derived from eggs deposited the previous autumn gave rise to the first infection of the animals in January and May. This infection had low consequences for the animals, but it signified the initiation of infection and the resumption of the annual cycle of pasture contamination. The second generation, appearing between June and July, was the year's most important source of infection and gave rise to an outbreak of parasitism in permanent lambs. This was indicated by an increase in levels of serum pepsinogen, the appearance of diarrhoea, and an important decrease in lamb growth rates. The third wave of infection, occurring in October and November, there was less impact than the previous one with respect to the level of infection, but it was the origin of the future over-wintering population. Teladorsagia circumcincta was the predominant species, followed by Haemonchus contortus and Trichostrongylus colubriformis. The highest numbers of worm recovered from both the tracer and permanent lambs were observed in July, with average numbers of 7900 and 19,200, respectively. The inhibition phenomenon was more evident in permanent than in tracer lambs, and mainly affected H. contortus as evidenced by populations exhibiting arrested larvae values of over 70% in January and February. Results obtained in this study confirm the epidemiological importance of the over-wintering L(3) generation as the initial source of the animal's infection and the origin of the annual resumption of pasture contamination cycles.

Animal Husbandry↗

[Stool behavior following the administration of antacids].

The direct correlation between the dosage of magnesium in antacids and the appearance of partly massive diarrheas in ulcus-patients known from literature could be confirmed. The clinically relevant dosage of antacids of 420 mval NK/day, divided in six individual dosages, resulted in diarrheas from the second to fourth day in 32 percent of the test persons when antacid A with a high content of magnesium was administered. There was no hint for a constipating effect of Solugastril gel at these high dosages (= 12 individual doses). Indeed the application of an Al-Ca-antacid may result in a slight fecal impaction, however, at the same time also to an increase of stool frequency, which is similar to that obtained with antacid B having a lower content of magnesium. The antacid can be regarded as neutral concerning stool frequency and consistency in the majority of test persons. From all four tested antacids the triple-combination (Al, Mg, Ca), showed the least important deviations from the preceding norms concerning stool frequency and consistency. The triple-combination shows an absolutely neutral stool behaviour. Serum electrolytes remained in the normal range with the Al-Ca- and the Al-Mg-Ca-antacid; whereas after application of antacid. A with high magnesium content, important increases of magnesium could be observed after seven days already, which were also dealt with in the literature.

Aluminum Hydroxide↗

Veterans Administration Cooperative Study on Bowel Preparation for Elective Colorectal Operations: impact of oral antibiotic regimen on colonic flora, wound irrigation cultures and bacteriology of septic complications.

A ten hospital cooperative study comparing prophylactic oral neomycin and erythromycin base versus placebo demonstrated clinical efficacy of the antibiotics in preventing septic complications following elective colon operations. The present report concerns microbiological studies accomplished during this trial. Cultures of colon contents during surgery showed the antibiotic prep reduced concentrations of both aerobes and anaerobes by approximately 10(5) bacteria/ml. Virtually all major bacterial components of the normal flora were affected. Wound irrigation specimens at the time of closure failed to predict subsequent wound infection, but significantly fewer antibiotic recipients had positive irrigation cultures. Postoperative stool specimens showed that the oral antibiotics did not cause an emergence in resistant forms. Bacteriological studies of postoperative infections indicated that most postoperative infections involved a mixed aerobic-anaerobic flora, and that Bacteroides fragilis accounted for six of eight episodes of bacteremia.

Bacteria↗

Three-dimensional modeling of fecal coliform in the Tidal Basin and Washington Channel, Washington, DC.

Fecal coliform are widely used as bacterial indicator in the United States and around the world. Fecal coliform impaired water is highly possible to be polluted by pathogenic bacteria. The Tidal Basin and Washington Channel in Washington, DC are on the Total Maximum Daily Load (TMDL) list due to the high fecal coliform level. To support TMDL development, a three-dimensional numerical model of fecal coliform was developed using the EFDC framework. The model calculates the transport of fecal coliform under the influences of flap gate operations and tidal elevation. The original EFDC code was modified to calculate the die-off of fecal coliform under the impact of temperature and solar radiation intensity. The watershed contribution is expressed as storm water inflow and the load carried by the runoff. Model results show that fecal coliform vary strongly in space in both the Tidal Basin and Washington Channel. The storm water only impacts a small area around the storm water outfall in the Tidal Basin and the impacts are negligible in the Washington Channel due to dilution. The water from the Potomac River may affect the fecal coliform level in the area close to the flap gate in the Tidal Basin. The fecal coliform level in the Washington Channel is mainly controlled by the fecal coliform level in the Anacostia River, which is located at the open boundary of the Washington Channel. The potential sediment layer storage of fecal coliform was analyzed and it was found that the sediment layer fecal coliform level could be much higher than the water column fecal coliform level and becomes a secondary source under high bottom shear stress condition. The developed model built solid connection of fecal coliform source and concentration in the water column and has been used to develop TMDL.

District of Columbia↗

[Functional dyspepsia. The physician and the patient].

Functional dyspepsia is defined by the presence of pain/discomfort in the upper abdomen without evident of organic disease which explain it; it must not be relief by defecation and its onset must not be related to changes in freqfrecuentlyuency or consistency feces. Diagnosis also requires pain or discomfort to be present for 12 weeks, not necessarily consecutives, in the last year. It is classified in three subgroups: ulcer-like, dysmotility-like and unspecified). Functional dyspepsia represents not only a diagnostic challenge but also a therapeutic problem, since no specific drug is available. An appropriate management of functional dyspepsia should consider patient's personality, diagnostic work-up, therapeutic alternatives and patient-doctor relationship. Many patients "live" their disease as the center of their life, sometimes creating great problems to physicians. Doctors should dedicate enough time to the patient, show interest in patient's problems, make a rigorous physical examination, and perform an appropriate work-up individualized for each patient. Regarding therapeutic decisions, it is important to reassure patients about the absence of organic disease. However it should be avoid to tell them that no disease exist at all; instead, the functional nature of the disease must be emphasized, explaining what and how upper GI tract is malfunctioning; Patients should know that doctor understand their symptoms are true symptoms; also, they must be informed bout the excellent outcome, without changes in life expectancy and a natural trend to improve with time; doctor should help patients to recognize that emotional situation have a great impact in disease course; and a therapeutic plan should be discussed and agreed with patients.

Dyspepsia↗

Incidence and clinical impact of fluoroquinolone-resistant Escherichia coli in the faecal flora of cancer patients treated with high dose chemotherapy and ciprofloxacin prophylaxis.

This study evaluated the susceptibility of Escherichia coli to quinolones in 72 stool samples collected from 31 patients with solid tumours who had undergone high dose chemotherapy (HDC) and peripheral blood stem-cell (PBSC) rescue with ciprofloxacin prophylaxis. Samples were obtained at admission, after completing prophylaxis and three months later. All E. coli strains isolated from baseline samples were susceptible to quinolones. Fluoroquinolone-resistant E. coli strains were isolated in 10 (32%) patients in the second sample. In eight of these patients isolates were susceptible 3 months later. No patient developed infection due to fluorquinolone-resistant E. coli. No differences were observed in outcome between patients with susceptible and resistant flora.

Adult↗

Use of selenite enrichment broth for the detection of Salmonella from stool: a report of one year experience at a provincial public health laboratory.

Several references recommend that selenite enrichment be used only in stool cultures from suspect carriers, during outbreaks, and in other special circumstances. To determine the impact of such an approach, we examined results from 3977 specimens cultured by our laboratory. Epidemiological information was collected from physicians and the public health department. Salmonella spp. were identified in 74 specimens from 54 patients. Four Shigella spp. were recovered from four patients. Forty-seven of the 74 Salmonella spp. were recovered on both the primary xylose-lysine-deoxycholate (XLD) and after enrichment. No Salmonella or Shigella grew on the primary XLD only. Twenty-six Salmonella spp. were recovered only after selenite enrichment. Of these, 17 were from newly identified patients. The elimination of selenite enrichment would have significantly reduced our yield, whereas the elimination of the primary XLD would not have resulted in any fewer isolates and would have resulted in savings of approximately $4000 yearly.

Carrier State↗

Prevalence of virulence genes and clonality in Escherichia coli strains that cause bacteremia in cancer patients.

Phenotypic analysis of Escherichia coli strains causing bacteremia in cancer patients suggests that they possess specific virulence properties. To investigate this hypothesis, we compared the frequency of the virulence-related genes cnf1, cnf2, papC, hlyC, and iut in 155 E. coli strains isolated from hospitalized cancer patients with epidemiologically unrelated cases of bacteremia to their frequency in 70 E. coli strains isolated from the feces of healthy unrelated volunteers. Of the blood isolates, 24, 37, and 26% were positive for cnf1, papC, and hlyC, respectively, versus only 6, 17, and 6% of the fecal isolates (P < 0.05 in all instances). By contrast, 47% of both isolates carried the iut gene. The patients' clinical characteristics did not significantly influence these frequencies. The presence on various pathogenicity islands (PAIs) of a combination of the cnf1, papC, and hlyC genes on the chromosome was strongly suggested by Southern blotting of pulsed-field gel electrophoresis (PFGE) patterns with specific DNA probes. The phylogenetic relatedness among 60 strains carrying three, two, one, or no virulence genes and 6 ECOR strains included as references was determined by neighbor joining, the unweighted pair-group method with arithmetic mean, and Wagner analysis of the randomly amplified polymorphic DNA (RAPD) patterns generated by 11 primers. Identification of a major cluster including 96.4% of the strains carrying the cnf1, papC, and hlyC genes and ECOR subgroup B2 strains suggested that the virulent E. coli strains causing bacteremia in cancer patients are closely related to ECOR B2 strains. The presence in the E. coli population surveyed of a strong linkage disequilibrium, and especially of a highly significant correlation between PFGE and RAPD genetic distances, confirms that clonal propagation has a major impact on the E. coli population structure. Nevertheless, low bootstrap values in the phylogenetic tree suggested that frequent genetic exchange inhibits the individualization of discrete genetic lineages, which are stable on an evolutionary scale.

Animals↗

The relative stability of chronic Isospora sylvianthina (Protozoa: Apicomplexa) infection in blackcaps (Sylvia atricapilla): evaluation of a simplified method of estimating isosporan infection intensity in passerine birds.

Estimation of intensity of coccidia infection in wild birds is essential for the studies of the parasite impact on birds' natural populations. In this paper I propose a method for quantification of coccidia oocyst output from passerine birds applicable in field investigations because it is based on one faeces droplet per bird only. This method was checked under controlled laboratory conditions on blackcaps, chronically infected with Isospora sylvianthina, and was proved to be sensitive enough for the detection of between-individual differences in parasite load. I also show that individual variation in the intensity of isosporan infection in captive blackcaps is relatively stable in time, both in the scale of a few consecutive hours and in that of consecutive days. Hence, the study shows that, in passerine birds, single measurements of oocyst production, when collected in the proper time of day, can be used to characterise some more permanent aspects of an individual's coccidia infection status.

Animals↗

Reducing fluxes of faecal indicator compliance parameters to bathing waters from diffuse agricultural sources: the Brighouse Bay study, Scotland.

The European Water Framework Directive requires the integrated management of point and diffuse pollution to achieve 'good' water quality in 'protected areas'. These include bathing waters, which are regulated using faecal indicator organisms as compliance parameters. Thus, for the first time, European regulators are faced with the control of faecal indicator fluxes from agricultural sources where these impact on bathing water compliance locations. Concurrently, reforms to the European Union (EU) Common Agricultural Policy offer scope for supporting on-farm measures producing environmental benefits through the new 'single farm payments' and the concept of 'cross-compliance'. This paper reports the first UK study involving remedial measures, principally stream bank fencing, designed to reduce faecal indicator fluxes at the catchment scale. Considerable reduction in faecal indicator flux was observed, but this was insufficient to ensure bathing water compliance with either Directive 76/160/EEC standards or new health-evidence-based criteria proposed by WHO and the European Commission.

Agriculture↗

Commercial soybean starch blocker consumption: impact on weight gain and on copper, lead and zinc status of rats.

A commercial 'starch blocker' was used to study the digestion of starch (potato) in mature female rates for four weeks. Two levels of 'starch blocker' were used. The first level was calculated to inhibit starch digestion at 50%, the second was calculated to inhibit starch digestion at 100%. No significant effects on the body weights (271.10 +/- 29, 277.7 +/- 43, 259.1 +/- 25 g) were found among the groups of rats at 0%, 50% and 100% inhibition levels, respectively. Feed intakes were not affected. However, fecal copper and zinc excretions increased significantly (p less than 0.05) due to the inhibitors. Fecal copper excretions were 0.468 +/- 0.14, 0.578 +/- 0.09, 0.617 +/- 0.07 mg/rat/week, while fecal zinc values were 0.625 +/- 0.14, 0.859 +/- 0.32 and 0.778 +/- 0.26 mg/rat/week when no inhibitor was fed, at 50% inhibition and at 100% inhibition respectively. Thus, while use of 'starch blockers' did not promote weight loss in the mature female rats, utilization of copper and zinc were negatively affected.

Amylases↗

Large Quantities of Bacterial DNA and Protein in Common Dietary Protein Source Used in Microbiome Studies.

Diet has been shown to greatly impact the intestinal microbiota. To understand the role of individual dietary components, defined diets with purified components are frequently used in diet-microbiota studies. Defined diets frequently use purified casein as the protein source. Previous work indicated that casein contains microbial DNA potentially impacting results of microbiome studies. Other diet-based microbially derived molecules that may impact microbiome measurements, such as proteins detected by metaproteomics, have not been determined for casein. Additionally, other protein sources used in microbiome studies have not been characterized for their microbial content. We used metagenomics and metaproteomics to identify and quantify microbial DNA and protein in a casein-based defined diet to better understand potential impacts on metagenomic and metaproteomic microbiome studies. We further tested six additional defined diets with purified protein sources with an integrated metagenomic-metaproteomic approach and found that contaminating microbial protein is unique to casein within the tested set as microbial protein was not identified in diets with other protein sources. We also illustrate the contribution of diet-derived microbial protein in diet-microbiota studies by metaproteomic analysis of stool samples from germ-free mice (GF) and mice with a conventional microbiota (CV) following consumption of diets with casein and non-casein protein. This study highlights a potentially confounding factor in diet-microbiota studies that must be considered through evaluation of the diet itself within a given study.

Animals↗

Bioluminescent bacteria assay of veterinary drugs in excreta of food-producing animals.

The residues of pharmacological treatments on food-producing animals, present in the manure dispersed on agricultural land, can impact environmental and human health through toxic, genotoxic, and drug-resistance development effects. Biotoxicity assays can easily reveal the presence of noxious substances and those based on bioluminescent bacteria (BLB) are particularly simple and rapid. A BLB assay was developed as microplate format by using various strains of Vibrio sp. and was employed to evaluate their response to pure antibiotic solutions and to residues extracted from excreta of antibiotic treated pigs and turkeys. The residues were quantified by HPLC analysis. The BLB assay can be proposed as an easy-to-perform screening tool to assess the presence of residues due to undeclared current, or recently ended, pharmacological treatments, as well as to evaluate their permanence in manure.

Animals↗