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PubMed1997

Flow cytometric detection of Cryptosporidium oocysts in human stool samples.

Cryptosporidium parvum is an important pathogen that causes diarrhea in virtually all human populations. Improved diagnostic methods are needed to understand the risk factors, modes of transmission, and impact of cryptosporidiosis. In the present study, we fluorescently labeled and counted C. parvum oocysts by flow cytometry (FC) and developed a simple and efficient method of processing human stool samples for FC analysis. Formed stool (suspended in phosphate-buffered saline) from an asymptomatic, healthy individual was seeded with known concentrations of oocysts, and oocysts were labeled with a cell wall-specific monoclonal antibody and detected by FC. The method described herein resulted in a mean oocyst recovery rate of 45% +/- 16% (median, 42%), which consistently yielded a fourfold increase in sensitivity compared to direct fluorescent-antibody assay of seeded stool samples. However, in many instances, FC detected as few as 10(3) oocysts per ml. Thus, FC provides a reproducible and sensitive method for C. parvum oocyst detection.

Animals↗

PubMed1993

Impact of organochlorine pesticide residues and PCBs on otters (Lutra lutra) in eastern England.

A study of otter (Lutra lutra) populations and levels of organochlorine (OC) pesticide residues and PCBs in otter faeces (spraints) was made on rivers in East Anglia, England. Population and contaminant levels are compared against target values. It is thought that the native otter population was almost extirpated during the study period. The current population must derive largely, if not entirely, from captive-bred animals introduced since 1983. Its range has expanded little and population levels, measured as an index, were generally low. Concentrations of OC pesticides and PCBs were similar to those of lowland stretches of western rivers; however, samples had a smaller proportion of dieldrin and a greater proportion of PCBs. Overall 44% of samples had concentrations of contaminants exceeding the 'level of concern'. It is considered that contamination, especially by PCBs, remains a factor influencing otter populations, which may not be viable in East Anglia without repeated releases of captive-bred animals.

Animals↗

PubMed2006

Induced biliary excretion of Listeria monocytogenes.

Listeria monocytogenes is a ubiquitous gram-positive bacterium that can cause systemic and often life-threatening disease in immunocompromised hosts. This organism is largely an intracellular pathogen; however, we have determined that it can also grow extracellularly in animals, in the lumen of the gallbladder. The significance of growth in the gallbladder with respect to the pathogenesis and spread of listeriosis depends on the ability of the bacterium to leave this organ and be disseminated to other tissues and into the environment. Should this process be highly inefficient, growth in the gallbladder would have no impact on pathogenesis or spread, but if it occurs efficiently, bacterial growth in this organ may contribute to listeriosis and dissemination of this organism. Here, we use whole-body imaging to determine the efficacy and kinetics of food- and hormone-induced biliary excretion of L. monocytogenes from the murine gallbladder, demonstrating that transit through the bile duct into the intestine can occur within 5 min of induction of gallbladder contraction by food or cholecystokinin and that movement of bacteria through the intestinal lumen can occur very rapidly in the absence of fecal material. These studies demonstrate that L. monocytogenes bacteria replicating in the gallbladder can be expelled from the organ efficiently and that the released bacteria move into the intestinal tract, where they pass into the environment and may possibly reinfect the animal.

Animals↗

PubMed1976

Model ecosystem evaluation of the environmental impacts of the veterinary drugs phenothiazine, sulfamethazine, clopidol, and diethylstilbestrol.

Four veterinary drugs of dissimilar chemical structures were evaluated for environmental stability and penchant for bioaccumulation. The techniques used were (1) a model aquatic ecosystem (3 days) and (2) a model feedlot ecosystem (33 days) in which the drugs were introduced via the excreta of chicks or mice. The model feedlot ecosystem was supported by metabolism cage studies to determine the amount and the form of the drug excreted by the chicks or mice. Considerable quantities of all the drugs were excreted intact or as environmentally short-lived conjugates. Diethylstilbestrol (DES) and Clopidol were the most persistent molecules, but only DES bioaccumulated to any appreciable degree. Phenothiazine was very biodegradable; sulfamethazine was relatively biodegradable and only accumulated in the organisms to very low levels. Data from the aquatic model ecosystem demonstrated a good correlation between the partition coefficients of the drugs and their accumulation in the fish.

Animals↗

PubMed2004

Efficacy of immunologic assays for the detection of Johne's disease in dairy cows fed additional energy during the periparturient period.

This study was designed to evaluate if the immunosuppression typically observed during the immediate periparturient period (3 weeks before and after calving) in dairy cows influences the effectiveness of diagnostic tests for the detection of Johne's disease; and, if providing additional energy to the cows during this period would minimize any immunosuppressive effects. Twelve dairy cows naturally infected with Mycobacterium paratuberculosis were fitted with rumen cannulas in late gestation and assigned to treatment groups: control, n = 6; or stuffed, n = 6. Cows in the control group were allowed to consume feed ad libitum. Cows assigned to the stuffed treatment group were also fed ad libitum but received additional total mixed ration by manually stuffing their rumens with refused feed to maintain a dry matter intake of 2% body weight/day before calving and 2.5% body weight/day after calving. Parturition had a significant impact on immune function with significant reductions in M. paratuberculosis-specific antibodies detected in the serum and milk regardless of treatment group. Similarly, in vitro immunoglobulin production was decreased at calving for both treatment groups. In addition, stuffing cows modulated cell-mediated immune function by reducing antigen-specific lymphocyte proliferation and interferon-gamma production after calving. Shedding of M. paratuberculosis in the milk was apparent in 58% (7/12) of cows after parturition with no difference noted between control and stuffed animals. Parturition had no major effect on fecal shedding of cows regardless of treatment. These data suggest that parturition had a significant effect on immune function parameters including diagnostic tests for paratuberculosis. Furthermore, providing additional energy to cows with Johne's disease did not preclude immunosuppressive effects during the periparturient period.

Animals↗

PubMed1982

Impact of different betalactam antibiotics on the normal human flora, and colonization of the oral cavity, throat and colon.

Phenoxymethylpenicillin was given orally in doses of 800 mg twice daily for seven days to six patients, and bacampicillin was given in doses of 400 mg three times per day for seven days to another six patients. Saliva, throat and faecal specimens were taken for cultivation of aerobic and anaerobic bacteria. Only small changes in the normal saliva and throat flora were observed, and no changes in the faecal flora were noticed during the observation period. Cefoxitin was administered parenterally in doses of 2 g at 6 h intervals for 12 h to six other patients. Pronounced changes in the colon flora occurred. Of the aerobic bacteria, enterobacteria decreased and cefoxitin-resistant enterococci increased in number; of the anaerobic bacteria, gram-negative rods decreased in number. At the end of the administration period, all cefoxitin-resistant strains decreased, and suppressed enterobacteria and bacteroides increased in number.

Adult↗

PubMed2003

Impact of sanitation and health education on intestinal parasite infection among primary school aged children of Sherpur, Bangladesh.

This study was carried out in 1999-2000 in the northern part of Bangladesh to determine the impact of sanitary latrine use and of health education on intestinal parasites in school-aged children. The children were between 5 and 13 years of age and stool samples revealed that more than half (53%) of the study sample was still infected with one or more intestinal parasites even after 4 years of intervention. Ascariasis was found to have the highest prevalence rate (36.2%) and hookworm the lowest (10.7%). Intestinal parasite infection was significantly lower (P < 0.05) among those who used a sanitary latrine and received health education. This result is consistent with observations that the effect of sanitation and health education is slow to develop. Concerted primary healthcare activities with community development efforts should be undertaken to improve the overall living condition of the people of this area to control this problem.

Adolescent↗

PubMed2003

Effects of a polymer-coated urea product on nitrogen metabolism in lactating Holstein dairy cattle.

The purpose of this study was to evaluate the impact of polymer-coated urea on nitrogen retention, rumen microbial growth, and milk production and composition. Coated urea (CU) that is more slowly hydrolyzed to ammonia than unprotected urea could potentially be used more efficiently by rumen microorganisms. Eight cows were offered each of three diets in a randomized crossover design. Each treatment period consisted of a 14-d adjustment period and a 5-d collection period. Diets were formulated to maintain milk production while reducing plasma urea nitrogen concentrations and urinary nitrogen excretion. Diets consisted of corn silage, mixed grass/legume haylage, chopped alfalfa hay, corn meal, protein, vitamin and mineral supplements, in a total mixed ration and fed ad libitum. The diets contained 17.9%, 18.1%, and 16.4% CP and 0, 0.77%, and 0.77% CU (dry matter basis) and are denoted as CP18-CU, CP18+CU, and CP16+CU, respectively. Individual feed intakes were measured, and total fecal, and urine collections were conducted. Cows were milked twice daily at 0500 and 1700 h, and the milk sampled for composition and milk urea N analysis. Dry matter intake averaged 23.5 +/- 0.2 kg/d and was not altered by diet. Also, milk fat and true protein were not altered by diet and averaged 3.72 and 3.07%, respectively. Milk yield was highest for diets CP18-CU and CP18+CU. Significant differences were observed in N intake and excretion in urine, feces, and milk between dietary treatments. Cows fed CP16+CU consumed 11% less N than in CP18-CU. Cows fed CP18+CU showed the highest excretion of N in urine, and together with CP16+CU, the lowest N excretion in feces. Nitrogen excretion in milk was lower for cows fed CP16+CU. Calculated N balance was not significantly different between diets nor was it significantly different from zero. Efficiency of N capture in milk protein as a function of N intake was higher for animals on CP16+CU. Urinary excretion of purine derivatives was not different between diets, and estimated microbial CP was also similar. Coated urea was not effective at reducing nitrogen excretion by dairy cattle.

Animals↗

PubMed2004

Impact of consumption of oligosaccharide-containing biscuits on the fecal microbiota of humans.

Human subjects consumed biscuits containing either galacto-oligosaccharides or fructo-oligosaccharides in a double-blinded, crossover study. The impact of supplementing the diet with three biscuits per day on the fecal microbiota was evaluated by selective culture of particular bacterial groups, measurement of beta-galactosidase activity, and nucleic acid-based analytical methods (PCR-denaturing gradient gel electrophoresis [PCR-DGGE] and fluorescent in situ hybridization). The composition of the bifidobacterial populations was monitored at the level of species (PCR-DGGE) and strains (pulsed-field gel electrophoresis of DNA digests), and representative cultures were tested quantitatively for their ability to use galacto-oligosaccharides. Technical improvements to DGGE analysis of the microbiota were made by the use of an internal standard that allowed valid comparisons of fragment staining intensities to be made between profiles, the use of S1 nuclease digestion to remove single-stranded DNA to facilitate cloning of DNA sequences cut from gels, and the extraction of RNA to be used as the template in reverse transcription-PCR-DGGE. RNA-DGGE profiles were markedly different (Dice's similarity coefficient, 58.5%) from those generated by DNA-DGGE. Neither the sizes of the bacterial populations nor the DNA-DGGE profiles of the microbiota were altered by the consumption of the biscuits, but the RNA-DGGE profiles were altered by the detection or increased staining intensity of 16S rRNA gene sequences originating from Bifidobacterium adolescentis and/or Colinsella aerofaciens in the feces of 11 of 15 subjects. beta-Galactosidase activity was elevated in the feces of some subjects as a result of biscuit consumption. Subjects differed in the ability of the bifidobacterial strains harbored in their feces to use galacto-oligosaccharides. Our observations suggest that a phylogenetic approach to analysis of the gut ecosystem may not always be optimal and that a more physiological (biochemical) method might be more informative.

Base Sequence↗

PubMed1991

Repeated community-based chemotherapy for the control of Schistosoma mansoni: effect of screening and selective treatment on prevalences and intensities of infection.

The impact of repeated selective chemotherapy on prevalences and intensities of infection with Schistosoma mansoni was evaluated in Gihungwe Transversals 1 and 2 (initial prevalence 60%) and Buhandagaza/Kizina (initial prevalence 35%), two village clusters in Burundi. Surveys were carried out at months -6, -3, 0, 3, 6, 9, 12, 24, and 36, with reference to the first intervention; treatment with praziquantel (40 mg/kg) was given at months 0, 12, 24, and 36 to subjects showing parasite eggs on a single 28-mg Kato slide. A second slide was examined for monitoring purposes only. Over the pre-intervention period, the overall prevalences and intensities remained relatively stable, but important increases were observed in specific groups. The cure rate three months after the first treatment in those treated was 73% (Gihungwe) and 83% (Buhandagaza/Kizina), but the prevalence at the community level was reduced only by 50% and 46%, respectively. Fifty-six percent and 79%, respectively, of the remaining positive cases had not been treated, largely because they were (falsely) negative at the screening. Reinfection occurred mainly in Gihungwe and in younger age groups. One year after the second treatment, prevalences and intensities were further reduced in Gihungwe only; one year after the third treatment prevalences were not reduced further in either village group. The final prevalence of infection was approximately 25%, with infections with an intensity of over 100 eggs per gram of feces approximately 5%, in all four villages. Over 80% of the remaining cases had not been treated at the previous intervention; the sensitivity of the screening method appears to be a major determinant of the outcome of repeated selective treatment.

Adolescent↗

PubMed2004

Waterfowl and the bacteriological quality of amenity ponds.

This study investigated the impact of waterfowl on the bacteriological quality of village ponds in East Yorkshire, north-east England. Water and sediment samples were collected from ponds with and without resident ducks and geese; faecal indicator and potentially pathogenic bacteria were assayed by membrane filtration and by selective enrichment. Escherichia coli, faecal streptococci and, to a degree, Clostridium perfringens were more abundant in ponds with waterfowl; Salmonella was isolated in June-August from the sediment of a pond with waterfowl. The results suggested that the bacteriological quality of village ponds might be adversely affected by waterfowl. All water samples from ponds with waterfowl had faecal indicators at higher concentrations than EU requirements for bathing waters. Although these ponds are not bathing waters we suggest skin contact and accidental ingestion of water should be avoided.

Animals↗

PubMed1986

Impact of cefbuperazone on the colonic microflora in patients undergoing colorectal surgery.

Cefbuperazone was given intravenously in a dose of 1 g at induction of anaesthesia, followed by a subsequent dose of 1 g 12 h after the first dose, to 10 patients undergoing colorectal surgery. A series of serum and faecal specimens were taken for analysis of cefbuperazone concentrations. Tissue samples from the gut wall were taken at surgery. The maximum serum concentrations (mean value 98.6 +/- 9.1 mg/l) during surgery were reached 30 min after cefbuperazone administration. The cefbuperazone concentration in the tissue samples varied between 7.3 and 40.5 mg/kg and the concentration in the faecal samples between 0 and 27.0 mg/kg. Faecal samples were also collected during the investigation period for cultivation of aerobic and anaerobic bacteria. The aerobic bacteria--streptococci, enterococci and enterobacteria--were suppressed significantly during the prophylaxis period. Among the anaerobic bacteria, cocci, bifidobacteria, eubacteria, lactobacilli, clostridia, fusobacteria and bacteroides decreased markedly during the same period. After four weeks the microflora was normalized in all patients. No postoperative infections occurred.

Adult↗

PubMed2006

Altered gut flora and environment in patients with severe SIRS.

BACKGROUND: The gut is considered an important target organ of injury after severe insult such as sepsis, trauma, and shock. The impact of bacterial translocation or mesenteric lymph on systemic inflammatory response and multiple organ damage has been investigated in animals, but dynamic changes in the gut flora and environment have not been fully clarified in critically ill patients. In the present study, we quantitatively evaluated changes in the gut microflora and environment in patients with severe systemic inflammatory response syndrome (SIRS). METHODS: Twenty-five patients with severe SIRS, who fulfilled the criteria for SIRS, had a serum CRP level >10 mg/dL, and were treated in the intensive care unit for more than 2 days, were included in our study. SIRS was a result of sepsis in 18 patients, trauma in 6, and burn in 1. A fecal sample was used for quantitative evaluation of microflora (bacterial counts of 10 key groups including Bifidobacterium and Lactobacillus) by plate or tube technique and of the gut environment (pH and 9 organic acids by high speed liquid chromatography). Data obtained from patients were compared with corresponding data from healthy volunteers. RESULTS: Analysis of fecal flora confirmed that patients with severe SIRS had significantly lower total anaerobic bacterial counts (especially 2-4 log fewer "beneficial" Bifidobacterium and Lactobacillus) and 2 log higher "pathogenic" Staphylococcus and Pseudomonas group counts than those of healthy volunteers. Concentrations of total organic acids (especially "beneficial" short-chain fatty acids such as acetic acid, propionic acid, and butyric acid) in the feces were significantly decreased in the patients, whereas pH was markedly increased. CONCLUSIONS: The gut flora and environment are significantly altered in patients with severe SIRS. Abnormal gut flora and environment may affect systemic inflammatory response after severe insult.

Adult↗

PubMed1999

Neonatal tetanus associated with topical umbilical ghee: covert role of cow dung.

BACKGROUND: Previous studies in Pakistan have shown that ghee (clarified butter) is commonly applied to umbilical wounds of neonates, and have documented that such applications are a risk factor for neonatal tetanus (NNT). In-use contamination of ghee with Clostridium tetani has been demonstrated, but mechanisms underlying the risk of ghee have been incompletely evaluated epidemiologically. METHODS: Detailed information on ghee usage, including fuels used to heat it, was obtained from cases of NNT (n = 229) and their matched controls (n = 687) from a population-based study of NNT in Punjab Province, Pakistan. Design variables were created to evaluate the impact of different fuel sources on risk of ghee applications. RESULTS: Nearly one-third of all infants had ghee applied, and it was nearly always heated before application to umbilical wounds of newborns. After controlling for all factors found to be significantly associated with NNT in conditional logistic regression, only ghee that had always been heated with dried cow dung fuel was significantly associated with NNT. Topical antimicrobials and ghee were never applied together. CONCLUSIONS: Ghee applications to umbilical wounds, when heated with 'clean' fuels, appear to pose no increased risk of NNT, although handling practices undoubtedly result in hazardous microbial contamination. In contrast, ghee heated with dung fuel was significantly associated with NNT. The effective promotion of topical antimicrobials might help reduce ghee use, since the intended purpose of each is to enhance healing.

Administration, Topical↗

PubMed1986

Natural history of Giardia infection of infants and children in rural Guatemala and its impact on physical growth.

Longitudinal data on Giardia excretion, diarrheal disease, and physical growth during the first 3 yr of life collected more than 20 yr ago in 45 Guatemalan children were analyzed. This report describes the natural history of this infection and estimates its effects on growth. All children had at least one Giardia infection, prevalence and incidence rates reaching 20.2% and 5.3%, respectively by the end of the third year. The mean number of Giardia infections per child increased from 0.7 in the first to 3.6 in the third year. More than 40% of these infections lasted 2-6 wk or more and were commonly associated with diarrhea. Weight velocity was significantly lower in the second year of life in Giardia-infected than in Giardia-negative children (p = 0.03). The duration of Giardia episodes and their association with diarrhea appeared to be the most important factors associated with growth disturbance. Although simultaneous infection with other enteropathogens occurred in many children, our findings suggest that Giardia infection may have independent deleterious effects on children's growth.

Body Height↗

PubMed2003

Evaluation of gastro-intestinal nematode parasite control strategies for first-season grazing cattle in Sweden.

A three-year grazing experiment (1998-2000) was conducted with first-season grazing cattle (FSGC) on improved pastures in central-eastern Sweden. Comparison was made between five groups with 10 calves in each group where four of these were set stocked and either (1) untreated, (2) ivermectin bolus treated, (3) subjected to biological control with the nematophagous fungus Duddingtonia flagrans, or (4) treated with a copper wire particle bolus. The fifth treatment was an evasive grazing strategy, whereby untreated calves were turned out onto pasture used by older cattle the previous year and then these calves were moved to silage aftermath in mid-July. To introduce low-levels of parasite infection to the experiment, each animal received a 'priming dose' of approximately 5,000 Ostertagia ostertagi and 5,000 Cooperia oncophora infective third stage larvae immediately prior to the start of the first grazing year of the trial. Results showed that efficient and sustainable parasite control of FSGC was possible to achieve without the use of anthelmintics by using turnout pastures that the previous year had been grazed by older cattle, in combination with a mid-July move to aftermath leys. Biological control also proved beneficial but the efficacy was impaired if high faecal egg counts coincided with rapid dung pat degradation due to heavy rainfall. No indication of parasite control was observed with the copper wire particle bolus. It was also demonstrated that the impact of gastrointestinal (GI) parasitism varied between years and that the level of overwintering contamination is important but likewise, is unpredictable. Although faecal egg counts in 1999 were low, due both to a delayed turnout and drought for the major part of the grazing season, deposited eggs successfully developed to infective larvae and overwintered in large numbers. The population of overwintered infective larvae at the time of turnout in early May played an important role in the course of infection in 2000 and resulted in an average 65 kg advantage of the ivermectin treated calves compared with the untreated calves.Thus, this three-year grazing experiment has emphasised the importance of subclinical gastrointestinal nematode infections in FSGC in Sweden. In addition, the study has shown that adequate parasite control may be achievable without the use of anthelmintics.

Animal Husbandry↗

PubMed2003

Methoxylation of desazadesferrithiocin analogues: enhanced iron clearing efficiency.

The impact of altering the octanol-water partition properties (log P) of analogues of desazadesferrithiocin, (S)-4,5-dihydro-2-(2-hydroxyphenyl)-4-methyl-4-thiazolecarboxylic acid, on the ligands' iron clearing properties is described. Increasing chelator lipophilicity can both substantially augment iron clearing efficiency in Cebus apella primates as well as alter the mode of iron excretion, favoring fecal over urinary output. The complications of iron overload are often associated with the metal's interaction with hydrogen peroxide, generating hydroxyl radicals (Fenton chemistry) and, ultimately, other related deleterious species. In fact, some iron chelators actually promote this chemistry. All of the compounds synthesized and tested in the current study are shown to be both inhibitors of the iron-mediated oxidation of ascorbate, thus removing the metal from the Fenton cycle, and effective radical scavengers.

1-Octanol↗

PubMed2001

Clinical severity of Norwalk virus and Sapporo virus gastroenteritis in children in Hokkaido, Japan.

OBJECTIVE: To clarify the clinical significance and etiologic impact of Norwalk virus (NV) and Sapporo virus (SV) in viral gastroenteritis in Japanese children. STUDY DESIGN: Two outbreaks each of NV gastroenteritis and SV gastroenteritis occurring in an infant home in Sapporo, Japan, as well as 95 hospitalized children with acute gastroenteritis were retrospectively evaluated using a 0- to 20-point clinical severity scoring system. RESULTS: The mean severity scores for NV and SV gastroenteritis outbreaks were 7.9 and 5.2, respectively, as compared with 8.4 for rotavirus A gastroenteritis that occurred in the same infant home. Among 95 hospitalized children with acute gastroenteritis, rotavirus A was detected in 47% followed by NV in 18%. SV was not found. CONCLUSION: Our data indicate that NV can cause severe gastroenteritis and is an important etiologic agent in hospitalized cases, whereas SV causes mild gastroenteritis in Japanese children.

Acute Disease↗