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Monitoring of fecal pollution in coastal waters by use of rapid enzymatic techniques.

Enzyme assays for 4-methylumbelliferyl-beta-D-galactopyranosidase and 4-methylumbelliferyl-beta-D-glucuronidase activities were used for rapid detection (25 min) of fecal water pollution and to determine the impact of sewage discharge in coastal waters. Two coastal areas were investigated: (i) an estuary characterized by a high degree of contamination downstream of a discharge from a sewage treatment plant and a low degree of water renewal and (ii) a fjord with a low degree of pollution and a high degree of water renewal. Statistical analysis showed that a global correlation curve could be used to estimate concentrations of culturable fecal coliform bacteria in the two coastal areas, although environmental factors important for cell physiology (e.g., salinity) varied at different sampling locations. The sensitivity limit for detection of 4-methylumbelliferyl-beta-D-glucuronidase activity corresponded to bacterial concentrations on the order of 10 to 100 CFU/100 ml. The 4-methylumbelliferyl-beta-D-galactopyranosidase assay was less sensitive because of a higher rate of substrate autohydrolysis. The detection limit corresponded to bacterial concentrations on the order of 100 to 1,000 fecal coliforms per 100 ml.

Environmental Monitoring↗

Morbidity from Schistosoma mansoni infections: an epidemiologic study based on quantitative analysis of egg excretion in two highland Ethiopian villages.

Although human infection with Schistosoma mansoni is known to be widespread in Highland Ethiopia, no information is available on its public health impact. A sample of the inhabitants of two typical villages in the endemic region north of Lake Tana were studied by clinical and quantitative parasitologic methods. One village had a moderately high prevalence (43.3%), while the second, chosen for comparison, had a very low prevalence (10.7%). No evidence of severe hepatosplenic disease was found in 343 persons examined. Symptons and mild hepatomegaly compatible with S. mansoni infection were common, and in the high prevalence village complaints of abdominal pain and blood in the stool were significantly related to S. mansoni egg counts. However, all signs and symptoms compatible with S. mansoni infection, including hepatomegaly, were of greater or similar frequency in the village with the low prevalence of infection. The findings of minimal morbidity are compatible with the level of intensity of infection found in the area and are consistent with the findings from morbidity studies based on quantitative egg counts in other areas of the world.

Adolescent↗

Influence of intestinal bacterial desulfation on the enterohepatic circulation of dehydroepiandrosterone sulfate.

Selective association of germ-free (GF) rats with dehydroepiandrosterone sulfate (DHEAS) desulfating bacteria allowed us to assess the exact impact of intestinal bacterial desulfation on the excretion and enterohepatic circulation of orally administered DHEAS. Germ-free rats selectively associated with the DHEAS-desulfating strain Peptococcus niger H4 (H4 rats) excreted 50% of the total label recovered within 17 h vs 21 h in GF rats and 13 h 23 min in conventional (CV) rats. Germ-free rats excreted 30% of the total label recovered via their urine. However, association of GF rats with the desulfating microorganism increased urinary excretion to 46%, comparable to the 45.5% found in CV rats. Fractionation of fecal label yielded 70% sulfoconjugated DHEAS and 2% unconjugated dehydroepiandrosterone in GF rats vs 5 and 77% in CV rats, and 55 and 14% in H4 rats, respectively. Our results demonstrate that the intestinal bacterial desulfation of DHEAS stimulated the enterohepatic circulation of DHEAS. This in turn increased the urinary excretion of label resulting in an accelerated elimination of labeled DHEAS from the body.

Animals↗

Development of a novel protocol for RT-multiplex PCR to detect diarrheal viruses among infants and children with acute gastroenteritis in Eastern Russia.

Viral gastroenteritis is one of the most common illnesses in humans worldwide and it has a great impact on people. Recently, we reported three RT-multiplex PCR assays termed A, B and C that could detect three groups of diarrheal viruses; group A, B and C rotaviruses and adenovirus [Phan et al., J Med Virol 2004; 74:173-9]; norovirus GI and GII, sapovirus and astrovirus [Yan et al., J Virol Meth 2003; 114:37-44]; enteroviruses, hepatitis A and E viruses and influenza A virus [Phan et al., Arch Virol 2005; 1175-85], respectively. In the present study, we developed a novel protocol with a small volume of reaction mixture for RT and PCR products (only 8 microl and 11 microl, respectively) that can amplify genomes of target viruses simultaneously. A total of 100 fecal specimens from infants and children with acute gastroenteritis in Birobiclzhan city, Eastern Russia, were collected during November 2003 and March 2004 and tested for the presence of those viruses by the novel RT-multiplex PCR protocol. Group A rotavirus was the most prevalent (67%), followed by norovirus GII (4%), group C rotavirus (1%), and sapovirus (1%). Interestingly, one fecal specimen turned out to be positive for hepatitis A virus. The sensitivity and specificity of RT-multiplex PCR assays with a novel protocol demonstrated a strong validation against the previously published RT-multiplex PCR. The findings clearly indicated that this novel protocol is simple and cost-effective to investigate the molecular epidemiology of acute gastroenteritis caused by diarrheal viruses. This report is the first, to our knowledge, detecting hepatitis A virus in feces from diarrheal infants and children in Eastern Russia.

Child↗

Impact of organochlorine pesticide residues and PCBs on otters (Lutra lutra): a study from western Britain.

A study of otter (Lutra lutra) populations and levels of organochlorine (OC) pesticide residues and PCBs in otter droppings (spraints) was made on eight stretches of river in three catchments in Wales and West Midland England. Population and contaminant levels were compared against target values. The otter populations (as measured by an index) remained stable on one stretch over 11 years, while they increased rapidly to equilibrium on four other stretches. The three lower stretches of rivers showed more erratic trends and on only one stretch did the index exceed, occasionally, the target value. Marking intensity at spraint sites was also lower at the three lowland sites. Concentrations of dieldrin, DDE and PCBs were significantly greater in spraint samples from the three lowland stretches. Over 50% of samples from the three lowland stretches had OC concentrations above the maximum allowable concentration, whereas most samples from the upland stretches had concentrations below the 'no effects level'. It is suggested that the colonization, by otters, of their former lowland range is inhibited by OC contamination of their food chain but, if contaminant levels can be reduced, spread will occur rapidly.

Animals↗

The cutworm Peridroma saucia (Lepidoptera: Noctuidae) supports growth and transport of pBR322-bearing bacteria.

Variegated cutworms were exposed to bean plants in microcosms sprayed with pBR322-carrying strains of Enterobacter cloacae, Klebsiella planticola, and Erwinia herbicola. The three bacterial species exhibited differential survival on leaves, in soil, and in guts and fecal pellets (frass) of the insects. High numbers of Enterobacter cloacae(pBR322) were detected in all samples, while the other species were unable to establish residence in the insect. To assess the impact of this colonization on site-to-site transport of microorganisms, larvae were fed plants that had been sprayed with the bacteria and then were transferred to uninoculated plants. Cutworms were efficient carriers of Enterobacter cloacae(pBR322), as indicated by its rapid appearance on uninoculated leaves and continued persistence in the insects for 3 days after transfer. Few Erwinia herbicola(pBR322) and K. planticola(pBR322) were obtained from larvae after transfer, although up to 10(3) CFU/g were detected in soil and on plants. Differences in bacterial survival and growth were confirmed by incubating frass overnight and observing the change in population numbers. The proportion of total samples showing at least a 25-fold increase during incubation was 68% for Enterobacter cloacae(pBR322), 39% for K. planticola(pBR322), and 0% for Erwinia herbicola(pBR322). Our results emphasize the role that cutworms and possibly other insects have in persistence and growth of microorganisms in the environment.

Animals↗

Tetracycline susceptibility of the ingested Lactobacillus acidophilus LaCH-5 and Bifidobacterium animalis subsp. lactis Bb-12 strains during antibiotic/probiotic intervention.

We investigated the effects of oral therapy with doxycycline, a tetracycline group antibiotic, on the gastrointestinal (GI) survival and tetracycline susceptibility of probiotic strains Lactobacillus acidophilus LaCH-5 and Bifidobacterium animalis subsp. lactis Bb-12. In addition, the influence of doxycycline therapy on the diversity of the predominant faecal microbiota was evaluated by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Faecal samples from the antibiotic group (receiving antibiotics and probiotics) and the control group (receiving probiotics only) were analysed for anaerobically and aerobically growing bacteria, bifidobacteria and lactic acid bacteria as well as for the dominant microbiota. Although doxycycline consumption did not have a large impact on GI survival of the probiotics, it had a detrimental effect on the bifidobacteria and on the diversity of the dominant faecal microbiota. A higher proportion of tetracycline-resistant anaerobically growing bacteria and bifidobacteria was detected in the antibiotic group than in the control group. Several antibiotic group subjects had faecal B. animalis subsp. lactis Bb-12-like isolates with reduced tetracycline susceptibility. This was unlikely to be due to the acquisition of novel tetracycline resistance determinants, since only tet(W), which is also present in the ingested B. animalis subsp. lactis Bb-12, was found in the resistant isolates. Thus, concomitant ingestion of probiotic L. acidophilus LaCH-5 and B. animalis subsp. lactis Bb-12 with the antibiotic did not generate a safety risk regarding the possible GI transfer of tetracycline resistance genes to the ingested strains.

Adult↗

Helminthic control on grazing ruminants and environmental risks in South America.

The control of ruminant gastrointestinal nematodes and ectoparasites, one of the major production health problems, are heavily reliant on the systematic and sometimes abusive use of anthelmintic drugs. Endectocides are the most frequently used drugs, having high potency against arthropods and nematodes. Their dung pat residues enhance the risk of adverse effects on non-target invertebrates and on the general grazing cattle ecosystem. In this scenario, our objective is to review current knowledge of the agro-environmental and biodiversity impact and risks of endectocides in South America. The effects of faecal drug residues of ivermectin and doramectin on dung colonising invertebrates and dung degradation have been reported in studies carried out in the temperate centre of Argentina and in the subtropical region of Brazil. The results from Argentina showed a depression in invertebrate colonisation and degradation of dung recovered from endectocide treated cattle during the autumn. Comparisons have shown that ivermectin and doramectin have similar adverse effects. A decrease of Coleoptera larvae, Diptera larvae, Staphylinidae, Collembola, Acari and dung specific nematodes was noted in pats from both endectocides. The results from Brazil showed that dipterous larvae, Polyphaga coleopteran larvae and adults and mites were significantly reduced in the ivermectin treated group. The disturbances that macrocyclic lactones can produce on non-targeted invertebrates and on their associated participation in dung degradation and soil element recycling, are unpredictable and can negatively influence biodiversity and the agricultural ecosystem sustainability.

Animal Husbandry↗

Environmental impact of ivermectin excreted by cattle treated in autumn on dung fauna and degradation of faeces on pasture.

The effect of ivermectin excreted in faeces of treated cattle on dung fauna and dung degradation on pasture during autumn was evaluated. Two groups of calves were used. One group was treated subcutaneously with ivermectin while the other remained as untreated control. Faeces deposited on 1, 3, 7, 14 and 21 days post-treatment (dpt) were removed on 1, 3, 7, 14, 21, 30 and 60 days post-deposition (dpd) and were used to determine the concentration of ivermectin and the percentage of organic matter and for the collection of colonising organisms. Samples from 1 and 3 dpt contained the highest drug concentration and percentage of organic matter compared to the control group (p<0.05). Faeces from the treated group showed lesser abundance and diversity of arthropods (p<0.05) than the control group. A reduction in numbers and diversity of dung fauna in faecal samples from treated animals was most remarkable at 1, 3 and 7 dpt, coinciding with the highest concentration of ivermectin and organic matter percentage.

Animals↗

The establishment, composition and severity of infection of gastro-intestinal parasites and their impact on productivity of Tswana kids in southern Botswana.

The presence of gastro-intestinal parasites in Tswana kids (n = 7) aged 1-3 weeks was studied for a period of 6 months at the Botswana College of Agriculture. The aims of this study were to find the time when they first contracted internal parasite infections, as well as to determine the severity of the infections and also its relation to production indicators (body mass and packed cell volume) of the kids as they grew older. The results indicate that they contracted coccidial and roundworm infections at approximately one month of age or immediately thereafter. The most prevalent internal parasite was coccidia, which occurred throughout the study period followed by roundworms and the least was the tapeworm, Moniezia expansa. Generally, the infection levels of all internal parasites were lower than the critical mean log (faecal oocyst/egg count + 1) of 3.3 inferred to cause reduced production in small stock. The correlation coefficients were all positive; 0.4-0.9 for individual internal parasites and production indicators, indicating that these internal parasites did not have any adverse effects on production. It was concluded that there was no need to treat kids of this age group for internal parasites.

Animals↗

Double blind, placebo controlled trial of two probiotic strains in interleukin 10 knockout mice and mechanistic link with cytokine balance.

BACKGROUND: Prophylactic efficacy against colitis following lactobacillus consumption in interleukin 10 (IL-10) knockout (KO) mice has been reported. Whether this applies equally to other probiotic strains is unknown, and the mechanism is unclear. AIMS: (1) To compare the effect of feeding Lactobacillus salivarius subspecies salivarius 433118 and Bifidobacterium infantis 35624 against placebo on enterocolitis, the intestinal microflora, and (2) to compare the systemic immunological response to in vitro microbial challenge in probiotic fed and control IL-10 KO mice. METHODS: Three groups of 10 IL-10 KO mice were fed fermented milk products containing Lb salivarius 433118 at 10(9) CFU/ml, B infantis 35624 at 10(8) CFU/ml, and unmodified milk, respectively, for 19 weeks. Faecal samples were taken at regular intervals to confirm gut transit, recovery of fed probiotics, and to assess the impact on the microflora. At sacrifice, the bowels were histologically scored. Cytokine production from Peyers' patches and splenocytes was measured in vitro by ELISA. RESULTS: Faecal recovery of probiotics was confirmed in all probiotic fed mice but not in controls. Colonic and caecal inflammatory scores were significantly decreased in both groups of probiotic fed mice (p<0.05). Proinflammatory cytokine production by Peyers' patches and splenocytes was significantly reduced in probiotic fed animals whereas transforming growth factor beta (TGF-beta) levels were maintained. CONCLUSION: Both Lactobacillus salivarius 433118 and Bifidobacterium infantis 35624 significantly attenuate colitis in this murine model. Attenuation of colitis is associated with a reduced ability to produce Th1-type cytokines systemically and mucosally, while levels of TGF-beta are maintained.

Animals↗

Dietary fibre: consensus and controversy.

Technological advances have reduced and refined man's plant food intake and consequently brought about an unprecedented decline in his consumption of dietary fibre (DF). The emergence of certain diseases selectively in regions which have been affected the most by this dietary change has led to an enhanced awareness of the functions of DF. DF is a heterogeneous group of substances which resist digestion by the endogenous enzymes of the human gut, although they are fermented to a substantial extent by the bacterial flora of the large intestine. Chemically, DF essentially consists of nonstarch polysaccharides and lignin, and its major constituents are cellulose, hemicelluose, lignin and pectin. The physiological effects of DF are attributable largely to its physicochemical properties. DF primarily affects gastrointestinal (GI) function; its effects are observable at all stages from ingestion through defaecation. It restricts caloric intake, shows gastric and small intestinal transit, and affects the activity of digestive enzymes and release of GI hormones. Its overall impact is to reduce apparent digestibility of nutrients marginally but consistently. In the large intestine, DF accelerates transit, supports bacterial growth and serves to hold water. As a result, the faecal weight and water content increase, and the transit time generally becomes shorter. Secondary to its GI effects, DF attenuates postprandial glycaemia and has long term effects on glucose tolerance and lipoprotein metabolism. These effects have important implications in the aetiopathogenesis of constipation and its sequelae including diverticulosis, cholesterol gallstones, colorectal cancer, obesity, diabetes mellitus and atherosclerosis. DF has traditionally been used therapeutically for constipation; now its use in diabetes is also well established. Our appreciation of the role of DF in human nutrition has undergone a major change in the last two decades. From a redundant constituent of plant foods, it has now moved to the position of an essential nutrient, the deficiency of which seems to have serious consequences.

Animals↗

Impact of a fluoroquinolone formulary change on acquisition of quinolone-resistant gram-negative bacilli.

Rising rates of fluoroquinolone resistance in bacteria have been associated with increased fluoroquinolone use. In vitro data show differences in potency among fluoroquinolone antibiotics against gram-negative bacteria and have led to the hypothesis that rates of selection of resistant microorganisms may be affected by the choice of the specific fluoroquinolone. Because clinical data to prove this hypothesis are lacking, the aim of the present study was to determine rates of acquisition of quinolone-resistant gram-negative bacilli (QRGNB) in the fecal flora of medical intensive care unit patients before and after a formulary change from ciprofloxacin to levofloxacin. Unadjusted rate ratios for acquisition of QRGNB were 1.09 (95%CI, 1.02-1.16) for each day of ciprofloxacin use and 1.01 (95%CI, 0.87-1.17) for each day of levofloxacin use. Following adjustment for other antibiotic use, enteral feeding, APACHE II score, and nursing home admission, neither ciprofloxacin nor levofloxacin use was associated with acquisition of QRGNB. In conclusion, a formulary change from ciprofloxacin to levofloxacin was not significantly associated with an increased risk of acquisition of QRGNB.

Anti-Bacterial Agents↗

Chimpanzee reservoirs of pandemic and nonpandemic HIV-1.

Human immunodeficiency virus type 1 (HIV-1), the cause of human acquired immunodeficiency syndrome (AIDS), is a zoonotic infection of staggering proportions and social impact. Yet uncertainty persists regarding its natural reservoir. The virus most closely related to HIV-1 is a simian immunodeficiency virus (SIV) thus far identified only in captive members of the chimpanzee subspecies Pan troglodytes troglodytes. Here we report the detection of SIVcpz antibodies and nucleic acids in fecal samples from wild-living P. t. troglodytes apes in southern Cameroon, where prevalence rates in some communities reached 29 to 35%. By sequence analysis of endemic SIVcpz strains, we could trace the origins of pandemic (group M) and nonpandemic (group N) HIV-1 to distinct, geographically isolated chimpanzee communities. These findings establish P. t. troglodytes as a natural reservoir of HIV-1.

Animals↗

Enhanced retrieval of DNA from human fecal samples results in improved performance of colorectal cancer screening test.

Colorectal cancer accounts for more than 10% of all cancer deaths but is curable, if detected early. We reported previously on a stool-based screening test in which DNA from stool samples is subjected to genome analysis; sensitivity of the test has been limited in part by inefficiency of retrieving DNA from stool. Our aim was to test the impact of a new purification method that would increase the yield of human DNA from stool. DNA from 86 cancer and 100 non-cancer subjects (diagnosed by colonoscopy) were purified from stool with a new method for DNA recovery based on sequence-specific capture with acrylamide gel immobilized capture probes as well as with a previously developed magnetic bead-capture procedure. The new purification method gives an average 5.4-fold increase in the quantity of human DNA that can routinely be retrieved from fecal samples. The increased recovery of DNA corresponds with an increase in assay sensitivity from 53% (CI: 42 to 64%) to 70% (CI: 59 to 79%); P = 0.0005 (by McNemar's test), with no change in specificity. The newly developed sample preparation method mitigates a major problem in detecting rare cancer-associated genetic changes in heterogeneous clinical samples such as stool.

Acrylamide↗

Transport of chemical and microbial compounds from known wastewater discharges: potential for use as indicators of human fecal contamination.

The quality of drinking and recreational water is currently (2005) determined using indicator bacteria. However, the culture tests used to analyze forthese bacteria require a long time to complete and do not discriminate between human and animal fecal material sources. One complementary approach is to use chemicals found in human wastewater, which would have the advantages of (1) potentially shorter analysis times than the bacterial culture tests and (2) being selected for human-source specificity. At 10 locations, water samples were collected upstream and at two successive points downstream from a wastewaster treatment plant (WWTP); a treated effluent sample was also collected at each WWTP. This sampling plan was used to determine the persistence of a chemically diverse suite of emerging contaminants in streams. Samples were also collected at two reference locations assumed to have minimal human impacts. Of the 110 chemical analytes investigated in this project, 78 were detected at least once. The number of compounds in a given sample ranged from 3 at a reference location to 50 in a WWTP effluent sample. The total analyte load at each location varied from 0.018 microg/L at the reference location to 97.7 microg/L in a separate WWTP effluent sample. Although most of the compound concentrations were in the range of 0.01-1.0 microg/L, in some samples, individual concentrations were in the range of 5-38 microg/L. The concentrations of the majority of the chemicals present in the samples generally followed the expected trend: they were either nonexistent or at trace levels in the upstream samples, had their maximum concentrations in the WWTP effluent samples, and then declined in the two downstream samples. This research suggests that selected chemicals are useful as tracers of human wastewater discharge.

Bacteria↗

The epidemiology of a recent focus of mixed Schistosoma haematobium and Schistosoma mansoni infections around the 'Lac de Guiers' in the Senegal River Basin, Senegal.

A village with mixed Schistosoma mansoni and S. haematobium infections (probably in a early endemic phase) was identified around the Lac de Guiers in the Senegal River Basin. In documenting the epidemiology of both schistosomes, we focused on prevalence and intensity of infection, transmission patterns and the impact of treatment. S. mansoni prevalences (near 100%) and egg counts (overall geometric mean eggs per gram of faeces (epg) of 589 were high in all age groups, with 35% of individuals excreting > 1000 epg, and showing a slow decline in egg output only after the age of 30 years. The overall prevalence (28%) and egg counts (2% > 50 eggs/10 ml) of S. haematobium were low, with mean counts of 6.3 eggs/10 ml. Maximal mean S. mansoni egg counts were found in 5-9 year-old boys and in 15-19 year-old girls; S. haematobium maximal counts in 1-4 year-old boys and in girls aged 5-9. Extremely high Biomphalaria pfeifferi infection ratios were recorded over the whole year. Following a single treatment, re-infection was rapid with prevalences and mean egg counts of both Schistosoma species reaching pretreatment levels within 7 months.

Adolescent↗

Infection management during antileukemia treatment-induced granulocytopenia: the role for oral norfloxacin prophylaxis against infections arising from the gastrointestinal tract.

Aerobic gram negative bacterial infections occur commonly in patients with acute leukemia undergoing intensive chemotherapy-induced bone marrow aplasia, and often arise from the gastrointestinal (GI) tract. The value of oral prophylactic norfloxacin (400 mg every 12 hours) in preventing bacterial infections was determined for 68 adults with acute leukemia by a prospective, randomized, double-blind, placebo-controlled trial. The efficacy of norfloxacin administered prophylactically throughout the course of intensive therapy and deep (less than 100 mm/3), prolonged (median 32 day) granulocytopenia against infections arising from the GI tract was demonstrated by its impact on the clinical manifestations of infection, microbiologically documented gram negative bacterial infections, overall antibiotic management, and GI colonization. Although there was no difference with respect to survival, norfloxacin decreased overall morbidity associated with gram-negative infections, was well tolerated, did not impinge on systemic antibacterial or antitumor therapy, and did not predispose to the development of bacteria resistant either to itself or to multiple other antibiotics. The advantages of norfloxacin relative to other oral agents used for GI prophylaxis and the potential beneficial effects of norfloxacin or overall medical cost are discussed. Oral norfloxacin achieves the goals of GI prophylaxis by effectively suppressing early infection arising from the GI tract and inhibiting GI colonization by multiply resistant pathogens that could give rise to late-onset infections during profound chemotherapy-induced bone marrow aplasia.

Acute Disease↗