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Presence of human Giardia in domestic, farm and wild animals, and environmental samples suggests a zoonotic potential for giardiasis.

Giardia lamblia which parasitize humans belong to either of two genotypes, A or B, based on specific signature sequences in the 5' end of the small subunit (16S) ribosomal RNA (rRNA) gene. These two genotypes also were found in cysts from fecal samples of animal origin such as dogs, cats, some farm animals and wild animals. In addition, trophozoites recovered from cysts obtained from environmental samples belonged to these two genotypes as well, suggesting that the G. lamblia genotypes A and B are widespread and possibly zoonotic. Trophozoites were recovered from rats and these isolates might belong to another genotype of G. lamblia. Deer mice and one dog appeared to be parasitized by genotypes of Giardia with close affinity to G. microti. This species, therefore, also consists of a genotype complex.

Animals↗

Effect of tigecycline on normal oropharyngeal and intestinal microflora.

Antibacterial agents disrupt the ecological balance of the normal human microflora. Tigecycline, a member of a new class of antibiotics (glycylcyclines), has been shown to have a potent broad-spectrum activity against most gram-positive and gram-negative aerobic and anaerobic bacteria. The aim of the study was to investigate the ecological effects of tigecycline on the normal oropharyngeal and intestinal microflora of healthy subjects. Thirteen healthy white subjects (six females and seven males) between 20 and 31 years of age received 100 mg of tigecycline in the morning on day 1 as a 30-min intravenous infusion followed by a 50-mg dose of tigecycline every 12 h as a 30-minute infusion for 10 days. One subject was withdrawn on day 2 because of an adverse event (urticaria). Serum, saliva, and fecal samples were collected before, during, and after administration for microbiological cultivation and for assays of tigecycline. All new colonizing bacteria were tested for susceptibility (resistance of > or =8 mg/liter) during the investigation period. The fecal concentrations on day 8 were from 3.0 to 14.1 mg/kg, with a mean value of 6.0 mg/kg and a median value of 5.6 mg/kg. The saliva concentrations were generally low (0 to 0.12 mg/liter). A minor effect on the oropharyngeal microflora was observed. The numbers of enterococci and Escherichia coli cells in the intestinal microflora were reduced at day 8 (P < 0.05), while those of other enterobacteria and yeasts increased. There was a marked reduction of lactobacilli and bifidobacteria (P < 0.05) but no impact on bacteroides. No Clostridium difficile strains were isolated. Two Klebsiella pneumoniae strains and five Enterobacter cloacae strains resistant to tigecycline were found on day 8.

Adult↗

Preliminary report on the application of the coagglutination test for rapid diagnosis of cholera in the upper Solimöes River area in the Brazilian Amazon region.

The conventional diagnosis of cholera depends on complex bacteriological procedures. Coagglutination is a simple, rapid, inexpensive and efficient technique for the presumptive diagnosis of cholera. Of 840 fecal samples from suspected cases of cholera examined at Tabatinga (State of Amazonas, Brazil) 31 (3.6%) were confirmed by culture and 29 of them were also positive by the coagglutination test performed directly on the fecal enrichment broth (alkaline peptone water). About 90% of the positive coagglutination results were obtained after 5-h incubation at 37 degrees C and the sensitivity, specificity and accuracy of the method were 93.5%, 99% and 98.8%, respectively. Relative to the culture results, coagglutination yielded two false-negative and eight false-positive results. The coagglutination test for cholera can provide a rapid and reliable tool for epidemiological studies and for the planning of more effective measures against cholera.

Agglutination Tests↗

An enrichment broth culture-duplex PCR combination assay for the rapid detection of enterotoxigenic Clostridium perfringens in fecal specimens.

An enrichment broth culture-duplex PCR combination assay was devised to identify Clostridium perfringens directly from fecal samples. The method consists of a combination of short enrichment of samples in selective media, DNA isolation, and performing duplex PCR using two pairs of primers which identify C. perfringens strains that harbor the virulence enterotoxin gene. Comparison of two selective enrichment media and two incubation temperatures showed that the reinforced clostridial medium with neomycin was better than the fluid thioglycollate medium with neomycin (p<0.001); and incubation at 37 degrees C vs 45 degrees C showed no statistically significant difference (p=0.238). The optimal short time for pre-enrichment culture was 4 hours. The developed assay was applied to detect phospholipase C (plc) and enterotoxin (cpe) genes for C. perfringens in feces inoculated artificially with enterotoxigenic C. perfringens. The method could detect both gene products in samples inoculated with a minimum of 10(4) CFU per ml. When the method was applied to detect enterotoxigenic C. perfringens in 198 diarrhea patients, C. perfringens was found in 121 samples; 7 out of 121 samples were positive for both plc and cpe (prevalence of 5.8%). These results indicate that the developed assay was a suitable method for the rapid and specific detection of enterotoxigenic C. perfringens directly in fecal specimens of diarrhea patients, which will assist epidemiological investigations of food poisoning outbreaks and quality control of food products.

Base Sequence↗

Effects of milbemycin oxime on adult hookworms in dogs with naturally acquired infections.

Previous work indicated that adult Ancylostoma caninum can be removed from experimentally infected dogs, using a formulation of milbemycin oxime at dosage of 0.5 mg/kg of body weight. To determine the efficacy of this treatment in dogs naturally infected with adult hookworms, 24 mixed-breed dogs with patent hookworm infections were purchased from an out-of-state vendor, and 6 male and 6 female dogs were assigned to either a control group or a group that would be treated. Dogs were treated 10 days after their arrival and were euthanatized 1 week after treatment. Beginning 3 days before treatment, fecal samples were collected daily from all dogs, and the number of Ancylostoma eggs per gram of dry weight of feces was determined from each sample. By 1 week after treatment, the mean number of eggs being passed by the treated dogs had dropped from 12,700 to 10 eggs/g of dried feces; there was no apparent change in fecal egg counts for dogs of the control group. At necropsy, the mean number of adult A caninum in dogs of the treated and control groups was 1.3 and 56, respectively; in these naturally infected dogs, efficacy of treatment was calculated to be 97.8%. The mean number of adult Trichuris vulpis recovered in dogs of the control and treated groups at necropsy was 24 and 0, respectively, which yielded treatment efficacy of 100%. Although Uncinaria stenocephala and Toxocara canis appeared also to be removed by use of this dosage, too few dogs were in the study to calculate meaningful efficacies.(ABSTRACT TRUNCATED AT 250 WORDS)

Ancylostomiasis↗

Why do African Americans get more colon cancer than Native Africans?

The incidence of colorectal cancer (CRC) is dramatically higher in African Americans (AAs) than in Native Africans (NAs) (60:100,000 vs. <1:100,000) and slightly higher than in Caucasian Americans (CAs). To explore whether the difference could be explained by interactions between diet and colonic bacterial flora, we compared randomly selected samples of healthy 50- to 65-y-old AAs (n = 17) with NAs (n = 18) and CAs (n = 17). Diet was measured by 3-d recall, and colonic metabolism by breath hydrogen and methane responses to oral lactulose. Fecal samples were cultured for 7-alpha dehydroxylating bacteria and Lactobacillus plantarum. Colonoscopic mucosal biopsies were taken to measure proliferation rates. In comparison with NAs, AAs consumed more (P < 0.01) protein (94 +/- 9.3 vs. 58 +/- 4.1 g/d) and fat (114 +/- 11.2 vs. 38 +/- 3.0 g/d), meat, saturated fat, and cholesterol. However, they also consumed more (P < 0.05) calcium, vitamin A, and vitamin C, and fiber intake was the same. Breath hydrogen was higher (P < 0.0001) and methane lower in AAs, and fecal colony counts of 7-alpha dehydroxylating bacteria were higher and of Lactobacilli were lower. Colonic crypt cell proliferation rates were dramatically higher in AAs (21.8 +/- 1.1% vs. 3.2 +/- 0.8% labeling, P < 0.0001). In conclusion, the higher CRC risk and mucosal proliferation rates in AAs than in NAs were associated with higher dietary intakes of animal products and higher colonic populations of potentially toxic hydrogen and secondary bile-salt-producing bacteria. This supports our hypothesis that CRC risk is determined by interactions between the external (dietary) and internal (bacterial) environments.

Africa↗

Comparative prevalence of the mercury resistance gene merA in human feces, food, and environmental water from Japan, Vietnam, and Ghana.

In this study, we investigated the prevalence and abundance of the mercury resistance gene merA in human feces, retail chicken meat, and environmental water samples collected from Japan, Vietnam, and Ghana. A real-time PCR assay developed in this study demonstrated high specificity toward merA sequences from more than 12 bacterial species. Using this assay, merA was detected in 6.8% of human fecal samples in Japan (n&#x2009;=&#x2009;29), in contrast to significantly higher rates observed in Vietnam (70.2%, n&#x2009;=&#x2009;47) and Ghana (97.4%, n&#x2009;=&#x2009;39). Similar geographic trends were evident in the chicken meat samples: 18.5% in Japan (n&#x2009;=&#x2009;27), 66% in Vietnam (n&#x2009;=&#x2009;91), and 90% in Ghana (n&#x2009;=&#x2009;10). Environmental water samples showed a consistently high merA detection rate across all countries (75-100%, n&#x2009;=&#x2009;21), with substantially higher gene copy numbers in Vietnam and Ghana than in Japan. merA was detected in some water samples, even when total mercury concentrations were below the detection limit, indicating that molecular detection may offer greater sensitivity than traditional physicochemical methods. Mercury-resistant bacteria were successfully isolated and cultured, and Citrobacter freundii was identified as the representative strain. Genomic analysis revealed that merA was located on an IncFIB plasmid, flanked by insertion sequences, suggesting its potential for horizontal gene transfer. These findings highlight merA as a promising biomarker for environmental mercury exposure and support the utility of fecal merA analysis as a proxy for assessing mercury-related public health risks.

Humans↗

Individual intake and antiparasitic efficacy of free choice mineral and fenbendazole in range calves.

The current study was conducted to assess the feasibility of fenbendazole (FB) administration to steers in a free choice mineral supplement. Provision of free choice FB reduces the need for handling of animals as well as decreases the level of animal parasitism. Two separate trials were conducted using 400 +/- 19 kg Holstein steers (n=14 and 17) during the months of July and August. Each steer was tagged with a unique electronic identification (EID) ear tag and randomly allocated into one of two groups. The tags worked in conjunction with a mineral feeder equipped with a load cell by registering the steer's EID number every time the animal entered the electromagnetic field. Individual daily mineral intake and feeding times were determined over two 8-day periods of non-medicated mineral (no FB), separated by a 14-day period of medicated mineral (0.55% FB). Fecal samples were collected at the beginning and end of each trial period and were analyzed for gastrointestinal nematode eggs and Giardia cyst. There was a consistently high level of attendance for the entire experimental period, with the exception of the first six days of the adaptation period. There were three preferential times for visiting the mineral feeder, approximately 07:00, 12:00 and 18:00 h. Individual daily mineral and FB intake was 229 +/- 27.21 g/day and 2 +/- 0.14 mg/kg BW/day, respectively, for the 14-day drug delivery period. The levels of fecal nematode eggs and Giardia cysts decreased significantly (<0.01) between pre- and post-sampling, with reductions of 92% for nematode eggs and 85% for Giardia cysts. Free choice medication for the control of gastrointestinal parasites is potentially effective, provided that the appropriate drug concentration, adaptation period, intake level and duration of treatment is utilized.

Administration, Oral↗

Intestinal parasites and bacteria of mountain gorillas (Gorilla beringei beringei) in Bwindi Impenetrable National Park, Uganda.

A survey in 1994 examined intestinal helminths and bacterial flora of mountain gorillas (Gorilla beringei beringei) in Bwindi Impenetrable National Park, Uganda. Parasites and bacteria were identified to genus in the feces of two groups of tourist-habituated and one group of non-tourist-habituated mountain gorillas. Eggs were identified as those of an anoplocephalid cestode, and nematode eggs representative of the genera: Trichuris, Ascaris, Oesophagostomum, Strongyloides, and Trichostrongylus. This is the first report of Ascaris lumbricoides-like eggs in mountain gorillas. Fecal samples (n=76) from all groups contained helminth eggs, with strongyle eggs and anoplocephalid eggs being the most common. Salmonella and Campylobacter were found in both gorilla groups. Regular long-term non-invasive fecal monitoring of the populations of mountain gorillas is essential for the prevention and identification of potential health threats by intestinal parasites and bacteria in this highly endangered subspecies.

Animals↗

Modulation of the fecal microbiota by the intake of a Lactobacillus johnsonii La1-containing product in human volunteers.

Lactobacillus johnsonii La1 (La1) is a probiotic strain capable of stimulating the immune system of the host and interfering with gastrointestinal pathogens. This study evaluates how the ingestion of different amounts of La1 influences the main bacterial populations of the fecal microbiota. Eight asymptomatic volunteers participated in the study. After a basal period, they ingested daily 100 mL of a product containing 10(8) CFU mL(-1) of La1 during the first week, 200 mL during the second week and 500 mL during the third week. Fecal samples were obtained at the end of each period and subsequently during 7 weeks. Lactobacilli were determined by culture on MRS agar and La1 colonies were confirmed by ERIC-PCR. The main populations of fecal bacteria were identified by FISH and flow cytometry. At baseline, 37.7% of the total fluorescent bacteria were Eubacterium rectale, 18.3% Fusobacterium prausnitzii, 13.2% Bacteroides, 8.6% Atopobium, 2.30%, Clostridium histolyticum, 2.05% Bifidobacterium and 0.95% Lactobacillus. Fecal excretion of La1 increased during the intake period and decreased during the post-ingestion period, so that no La1 was observed in the stools of the volunteers seven weeks after the intake product has been finished. La1 intake increased the populations of C. histolyticum (p=0.049), Lactobacillus (p=0.056) and Bifidobacterium (p=0.067), and decreased those of F. prausnitzii (p=0.005) while it did not affect Bacteroides, E. rectale and Atopobium populations. These bacterial populations returned to their baseline levels during the post-ingestion period. The regular intake of a La1-containing product beneficially affects the homeostasis of the human fecal microbiota, probably contributing to the health-promoting effects of this probiotic.

Administration, Oral↗

Monitoring and survival of Lactobacillus paracasei LTH 2579 in food and the human intestinal tract.

A PCR-based detection system specific for Lactobacillus paracasei LTH 2579 was developed and applied to follow the fate of the strain in complex ecosystems. This strain was isolated from fruit mash and was characterised as being highly resistant to low pH and bile at concentrations as they occur in the human digestive tract. The application of the subtraction hybridisation technique permitted to identify a 235 bp chromosomal DNA fragment of strain LTH 2579. Based on this target sequence a specific PCR system was developed and combined with the species-specific PCR system for L. paracasei. This combination of PCR based detection systems was successfully applied to monitor L. paracasei LTH 2579 in fermented sausages which were inoculated with this strain (2.0 x 10(7) CFU/g) together with the strongly competitive L. sakei LTH 681 (1.0 x 10(6) CFU/g). At the time of consumption of the sausages the respective counts were 1.8 x 10(7) and 1.4 x 10(8) CFU/g. After consumption of the sausages by three volunteers L. paracasei LTH 2579 was recovered from fecal samples. The counts determined for the strain ranged between 1.2 x 10(7) and 1.5 x 10(8) CFU/g of feces. The fortuitous lactobacilli constituted a share of 5-12% of the lactobacilli in the fecal flora.

DNA Primers↗

Metabolism of the food-associated carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine by human intestinal microbiota.

2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine is a putative human carcinogenic heterocyclic aromatic amine formed from meat and fish during cooking. Although the formation of hazardous PhIP metabolites by mammalian enzymes is well-documented, nothing is known about the PhIP transformation potency of human intestinal bacteria. In this study, the in vitro metabolism of PhIP by human fecal samples was investigated. Following anaerobic incubation of PhIP with stools freshly collected from six healthy volunteers, we found that PhIP was extensively transformed by the human intestinal bacteria. HPLC analysis showed that the six human fecal microbiota transformed PhIP with efficiencies from 47 to 95% after 72 h incubation, resulting in one major derivative. ESI-MS/MS, HRMS, 1D (1H, 13C, DEPT) and 2D (gCOSY, gTOCSY, gHMBC, gHSQC) NMR, and IC analysis elucidated the complete chemical identity of the microbial PhIP metabolite as 7-hydroxy-5-methyl-3-phenyl-6,7,8,9-tetrahydropyrido[3',2':4,5]imidazo[1,2-a]pyrimidin-5-ium chloride. At present, no information is available about the biological activity of this newly discovered bacterial PhIP metabolite. Our findings however suggest that bacteria derived from the human intestine play a key role in the activation or detoxification of PhIP, a digestive fate ignored so far in risk assessments. Moreover, the variation in transformation efficiency between the human microbiota indicates interindividual differences in the ability to convert PhIP. This may predict individual susceptibility to carcinogenic risk from this suspected dietary carcinogen.

Adult↗

Escherichia coli K5 antigen in relation to various infections and in healthy individuals.

A phage specific for Escherichia coli K5 antigen was used to determine the frequency of K5 in strains of E. coli isolated from cases of sepsis, meningitis, and urinary tract infection (pyelonephritis, cystitis, and asymptomatic bacteriuria), as well as from fecal samples of healthy subjects. Although entirely absent from meningitic strains, K5 was found to be one of the most common E. coli capsular antigens, especially in strains causing sepsis. K5 was less common in the fecal strains of healthy subjects than in strains causing various urinary tract infections, between which there was no significant difference in its frequency. Thus, even if K5 is a less important antigen than K1 in the etiology of acute pyelonephritis or acute meningitis, as the fifth most commonly occurring K antigen in E. coli strains causing urinary tract infection it should be included as a component in any K antigen-based vaccine against acute pyelonephritis.

Antigens↗

Prevalence of Giardia intestinalis infection in dogs of breeding kennels in Japan.

Giardia intestinalis antigen in fecal samples was examined in 361 dogs of 14 breeding kennels located at various areas in Japan, using a commercial enzyme-linked immunosorbent assay (ELISA) kit. G. intestinalis antigen was detected in 37.4% of the fecal specimens. All of the 14 breeding kennels were positive for G. intestinalis antigen with the range from 6.7 to 59.3%. The prevalence in puppies (54.5%) was significantly (p < 0.01) higher than that in adults (30.9%). There was no difference in prevalence between males and females, and between the puppies from the mother dogs positive and negative for Giardia antigen. In conclusion, G. intestinalis widely invaded the breeding kennels in Japan.

Age Factors↗

Intestinal parasites of baboons (Papio cynocephalus anubis) and chimpanzees (Pan troglodytes) in Gombe National Park.

A survey of gastrointestinal parasites of nonhuman primates in Gombe National Park, Tanzania, was conducted during July and August 1989. Fresh fecal samples collected from 35 baboons (Papio cynocephalus anubis) and 20 chimpanzees (Pan troglodytes) were analyzed for intestinal parasites. Parasite identifications were based upon the size and appearance of eggs and larvae on both direct and fecal flotation examinations. Seven species of helminths and three species of protozoa were noted in the chimpanzees, and seven species of helminths, two species of trematodes, and one species of protozoa were noted in baboons. An unidentified parasite, with a thick egg shell similar to that of ascarids, was noted in both the chimpanzee and baboon population. Unfortunately a definitive identification of this parasite could not be made based upon egg size and morphology alone and insufficient numbers of larva were present to aid identification. One previously unreported parasite, Schistosoma mansoni, was identified in the baboon population. This is the first report of a schistosome in Gombe's nonhuman primate population.

Animals↗

[Effects of loperamide on the fecal flora in children in severe diarrheas].

Loperamide has been recently indicated in the management of infants with severe protracted diarrhea. The purpose of this study was to determine the effect of loperamide on fecal flora in severe protracted diarrheas. 19 children aged 1 to 36 months, with severe protracted diarrhea were studied: 14 received loperamide (0.5 mg/kg/d) and 5 were without loperamide treatment. Criteria analysed were: clinical tolerance (vomiting and abdominal distention); efficacy (number of stools, transit time and Na+/K+ in stools) and effect on fecal flora, with differential qualitative and quantitative analysis method (complete identification and counts of aerobic and strict anaerobic bacteria in fresh fecal samples before and 4 to 8 days after the beginning of loperamide). Parenteral and/or oral alimentation remain constant during the study. Results show a rapid (24 h) efficacy in 9/14. Although important changes in specific fecal flora counts was noticed for streptococcus D and Proteus as compared to five controls, no bacterial overgrowth appeared or was increased during loperamide treatment.

Child, Preschool↗

Prevalence and risk factors for protozoan and nematode infections among children in an Ecuadorian highland community.

In order to create a parasite prevention program for Santa Ana, Ecuador, we worked with local physicians to identify the prevalence and risk factors for intestinal parasitic infection among children. Two hundred households were randomly selected and asked to submit three stool samples from a selected child. Information from direct observation and questionnaires was used to determine environmental and behavioral risk factors for parasitic infection. Prevalence of any parasitic infection was 65.6% of the 189 children who submitted fecal samples. Parasitic infections included Entamoeba histolytica/E. dispar (46.6%), Giardia lamblia (18.5%) and Ascaris lumbricoides (20.1%). In a multivariate logistic model, risk factors (P<0.05) for E. histolytica/E. dispar included >1 month since antiparasitic treatment, not adding chlorine to drinking water, children not washing their hands before eating, and having a fewer number of electrical appliances in the home. The risk factor identified for G. lamblia was children not washing their hands before eating. Risk factors for A. lumbricoides included >6 months since antiparasitic treatment and having a fewer number of electrical appliances in the home. This study may have implications for other communities throughout the developing world.

Age Distribution↗

Fructooligosaccharides and fiber partially prevent the alterations in fecal microbiota and short-chain fatty acid concentrations caused by standard enteral formula in healthy humans.

The intestinal microbiota are important during enteral tube feeding because they exert colonization resistance and produce SCFAs. However, the effect of the enteral formula composition on major bacterial groups of the microbiota has not been clearly defined. The aim of this study was to investigate the effect of enteral formulas with and without prebiotic fructooligosaccharides (FOS) and fiber on the fecal microbiota and SCFAs. Healthy subjects (n = 10; 4 men, 6 women) consumed both a standard enteral formula and one containing FOS (5.1 g/L) and fiber (8.9 g/L) as a sole source of nutrition for 14 d in a randomized, double-blind, crossover trial with a 6-wk washout phase. Fecal samples were collected at the start and end of each formula phase, and were analyzed for major bacterial groups and SCFA concentrations using fluorescent in situ hybridization and GLC, respectively. Although there were reductions in total fecal bacteria due to both formula treatments, concentrations were higher after the FOS/fiber formula period compared with the standard formula period (11.2 +/- 0.2 vs. 11.0 +/- 0.2 log(10) cells/g, P = 0.005). The FOS/fiber formula increased bifidobacteria (P = 0.004) and reduced clostridia (P = 0.006). Compared with the standard formula, the FOS/fiber formula resulted in higher concentrations of total SCFA (332.4 +/- 133.8 vs. 220.1 +/- 124.5 micromol/g, P = 0.022), acetate (219.6 +/- 96.3 vs. 136.8 +/- 74.5 micromol/g, P = 0.034) and propionate (58.4 +/- 37.4 vs. 35.6 +/- 25.5 micromol/g, P = 0.02). This study demonstrates that standard enteral formula leads to adverse alterations to the fecal microbiota and SCFA concentrations in healthy subjects, and these alterations are partially prevented by fortification of the formula with FOS and fiber.

Bacteria↗