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Lungworms (Dictyocaulus arnfieldi): prevalence in live equids in Kentucky.

Prevalence of natural infections of the lungworm, Dictyocaulus arnfieldi, was investigated in Kentucky in 5,437 live equids (horses, ponies, and asses) on 91 farms by examination of fecal samples over a 15-month-period from January 28, 1983 to April 19, 1984. For the 91 farms investigated, asses only were examined on 1 farm; for the other 90 farms, examinations were made of horses on all of them, of ponies on 4 farms, and of asses on 9 farms. Larvae of D arnfieldi were found in fecal samples of 112 (2%) of 5,379 horses on the 90 farms of which 38% had greater than or equal to 1 infected animal; none of 19 ponies examined was infected. For 39 asses (donkeys and mules) examined, 21 (54%) were infected; infected asses were present on 80% of the 10 farms on which they were examined. The equids, excluding asses, that were examined and the percentage (in parentheses) infected with D arnfieldi included 4,329 (2%) Thoroughbreds, 684 (2%) Standardbreds, 78 (0%) American Saddle Horses, and 307 (3%) miscellaneous (other breeds or mixed breeds of horses and ponies). Equids were separated into 3 categories relative to previous treatment with ivermectin: A (no treatment), B (treatment), and C (treatment unknown).(ABSTRACT TRUNCATED AT 250 WORDS)

Age Factors↗

Taenia saginata: differential diagnosis of human taeniasis by polymerase chain reaction-restriction fragment length polymorphism assay.

Speciation of Taenia in human stool is important because of their different clinical and epidemiological features. DNA analysis has recently become possible which overcomes the problems of differentiating human taeniid cestodes morphologically. In the present study, we evaluated PCR coupled to restriction fragment length polymorphism to differentiate Taenia solium from Taenia saginata eggs present in fecal samples from naturally infected patients. A different DraI-RFLP pattern: a two-band pattern (421 and 100 bp) for T. saginata and a three-band pattern (234, 188, and 99 bp) for T. solium was observed allowing the two species to be separated. The lower detection limit of the PCR-RFLP using a non-infected fecal sample prepared with a given number of T. saginata eggs was 34 eggs in 2 g stool sediment. The 521 bp mtDNA fragment was detected in 8 out of 12 Taenia sp. carriers (66.6%). Of these, three showed a T. solium pattern and five a T. saginata pattern.

Animals↗

Detection of Helicobacter species DNA by quantitative PCR in the gastrointestinal tract of healthy individuals and of patients with inflammatory bowel disease.

In many animal species different intestinal Helicobacter species have been described and a few species are associated with intestinal infection. In humans, the only member of the Helicobacter family which is well described in literature is Helicobacter pylori. No other Helicobacter-associated diseases have definitely been shown in humans. We developed a sensitive quantitative PCR to investigate whether Helicobacter species DNA can be detected in the human gastrointestinal tract. We tested gastric biopsies (including biopsies from H. pylori positive persons), intestinal mucosal biopsies and fecal samples from healthy persons, and intestinal mucosal biopsies from patients with inflammatory bowel disease (IBD) for the presence of Helicobacter species. All gastric biopsies, positive for H. pylori by culture, were also positive in our newly developed PCR. No Helicobacter species were found in the mucosal biopsies from patients with IBD (n = 50) nor from healthy controls (n = 25). All fecal samples were negative. Our study suggests that Helicobacter species, other than H. pylori, are not present in the normal human gastrointestinal flora and our results do not support a role of Helicobacter species in IBD.

Biopsy↗

Molecular characterizations of human and animal group a rotaviruses in the Netherlands.

To gain more insight into interspecies transmission of rotavirus group A, human and animal fecal samples were collected between 1997 and 2001 in The Netherlands. A total of 110 human stool samples were successfully P and G genotyped by reverse transcriptase PCR. All strains belonged to the main human rotavirus genotypes G1 to G4, G9, [P4], [P6], [P8], and [P9]. [P8]G1 was predominant, and 5.5% belonged to the G9 genotype. Eleven percent of all P[8] genotypes could be genotyped only by a recently published modified primer. Rotavirus-positive fecal samples from 28 calf herds were genotyped by DNA sequencing. Genotypes G6 and G10 predominated; G6 and G10 were detected in 22 (78.6%) and 16 (57.1%) of the rotavirus-positive calf herds, respectively. In 12 (42.9%) calf herds, we found mixed infections. Genotype G8 was not found. Genotype G6 bovine rotaviruses were divided into three clusters: UK-like, VMRI-29-like, and Hun4-like. DNA sequencing of a part of the VP7 gene was shown to be useful as a quick determination of uncommon or novel strains of which the genotyping cannot be done by genotyping PCR. Of equine strains, both VP4 and VP7 genes could be used for genotyping: two [P12]G3 and four [P12]G14 equine rotaviruses were determined. We did not find indications for rotavirus interspecies transmissions, although the recently published human G6-Hun4 is genetically related to our G6 bovine isolates. All bovine, porcine, and equine rotaviruses were within genotypes previously reported for these animal species.

Animals↗

Presence and persistence of intestinal parasites in canine fecal material collected from the environment in the Province of Chubut, Argentine Patagonia.

We investigated the presence of intestinal parasites in canine feces collected from public squares in Comodoro Rivadavia, Chubut, Argentina (45 degrees S, 68 degrees W) and determined the persistence of Echinococcus granulosus eggs in those droppings under natural environmental conditions in that region. In the first experiment, we analyzed 163 fecal samples collected from urban squares during 8 months time and found parasitic elements in 46.6%. The presence of parasites was independent of the condition of the feces (fresh or dried; P>0.05). Parasites potentially pathogenic in man were present, such as Toxocara species (spp.), Taenia spp./Echinococcus spp., Uncinarias spp., and Entamoeba spp. In the second experiment, we analyzed two canine fecal samples contaminated with E. granulosus eggs, deposited for 41 months within the natural environment. These parasitic elements persisted during the entire study as attested by light microscopy and the ELISA coproantigen test. We propose the study of the presence of intestinal parasites in canine feces within the environment as a general strategy for identifying and monitoring areas of risk for canine-related zoonoses since we were able to demonstrate the persistence of E. granulosus eggs in deposited canine feces for over 3 years within the area studied.

Animals↗

Selection of Trypanosoma cruzi clonal genotypes (clonet 20 and 39) isolated from Bolivian triatomines following subculture in liquid medium.

Previous studies showed that two groups of Trypanosoma cruzi clonal genotypes named clonet 20 and clonet 39 were predominant in Triatoma infestans, the unique vector of Chagas disease in Bolivia. These groups of clones correspond to distinct genetic clusters. These clonets were detected in T. infestans and Rhodnius pictipes fecal samples before isolation and after culture by kDNA PCR (polymerase chain reaction) and hybridization of the amplified products with clonet specific kDNA probes named 20 and 39 as previously reported. Forty eight T. infestans and three R. pictipes infected insects captured at random in different Bolivian departments were proceeded. As previously reported the direct identification of the two major clonets in fecal samples allowed the detection of abundant mixed infections: 41% in the original sample, however after culture, only 6% of mixed infections were detected. Among the 21 parasite stocks isolated from digestive tracts where mixed infections were initially detected (clonet 20 + 39) clonet 20 alone was detected in 81% of them. This result clearly showed that the culture step selected clonet 20 parasites over those belonging to clonet 39. The taxonomic status of the isolated stocks was also confirmed by isoenzyme typing, and correlation was observed between clustering topology and hybridization patterns with the probes 20 and 39.

Animals↗

Prevalence of enteric pathogens in dogs of north-central Colorado.

To evaluate the prevalence of enteric pathogens in dogs of north-central Colorado, fecal samples were obtained from client-owned dogs presented to the Veterinary Teaching Hospital at Colorado State University for evaluation of acute small-bowel, large-bowel, or mixed-bowel diarrhea (n=71) and from age-matched, client-owned, healthy dogs (n=59). Infectious agents potentially associated with gastrointestinal disease were detected in 34 of 130 (26.1%) fecal samples. Agents with zoonotic potential were detected in feces from 21 (16.2%) of 130 dogs and included Giardia spp. (5.4%), Cryptosporidium parvum (3.8%), Toxocara canis (3.1%), Salmonella spp. (2.3%), Ancylostoma caninum (0.8%), and Campylobacter jejuni (0.8%). Positive test results occurred in dogs with or without gastrointestinal signs of disease. Dogs, particularly those in homes of immunocompromised humans, should be evaluated for enteric zoonotic agents.

Ancylostoma↗

[Incidence of zoonoses in petting zoos and evaluation of hygiene measures to prevent the transmission to humans].

In summer 2003, a study was performed in thirty Swiss petting zoos with the objective to determine the prevalence of zoonotic agents, and to describe hygiene measures implemented to reduce the risk of human infection. Fecal samples from different animal species were collected from the floor of pens to determine the prevalence of Salmonella spp., Campylobacter spp., verocytotoxin producing E. coli/ VTEC and Francisella tularensis. A questionnaire on hygiene measures, number of animals per species, housing system, care procedures and feeding was administered to every petting zoo to estimate exposure of visitors to zoonotic microorganisms. In total, 423 fecal samples were examined. Of these samples, 41 were positive for Campylobacter spp., which were mainly isolates from pigs and poultry (35% positive samples from each species). In pigs, 50% of the positive samples (6 samples) were typed as C. jejuni. The others were typed as C. coli (3) and C lan' (3), respectively. Five poultry isolates were typed as C. jejuni, and two as C. coli. Two samples were positive for Salmonella spp. Salmonella typhimurium was isolated from a goat, the other isolate could not be identified by serotyping. Neither Francisella tularensis nor verocytotoxin producing E. coli/ VTEC were found. The low prevalence of zoonotic microorganisms in Swiss petting zoos could be attributed to the cleanness of enclosures and animals, low stocking rates and good animal care. However, there is room for improvement concerning visitors' information on hygiene and hand washing. Furthermore, a strict separation between picnic - areas and animals should be enforced.

Animal Husbandry↗

[Surveillance finding on rotavirus in Changchun children's hospital during July 1998-June 2001].

OBJECTIVE: To establish baseline patterns of rotavirus diarrhea and to describe its epidemiologic features in Changchun city, prior to rotavirus vaccine immunization. METHODS: Hospital-based surveillance was conducted among children under 5 years old with acute diarrhea in Changchun Children's Hospital. Fecal samples were determined to identify rotavirus by PAGE and/or ELISA. G serotypes of rotavirus were identified by ELISA and/or nested RT-PCR. P genotyping were carried out by RT-PCR. All data were computerized and analysed by "Generic Manual on Rotavirus Surveillance" set by CDC in the USA. RESULTS: In total, 2 343 diarrhea cases were screened and 1 211 fecal samples were collected. Rotavirus was detected in 31.0% among outpatients and 52.9% in inpatients. During the peak of the season (November through March), 58.6% of diarrhea was caused by rotavirus among inpatients. 95.0% of rotavirus diarrhea cases occurred among children aged < 2 years. The predominant strain was serotype G1 (82.4%), followed by G2 (5.0%), G3 (3.3%), G4 (0.9%). P genotyping showed that P[8] and P[4] were the most common ones. Nine different P-G combinations were identified, four strains (P[8]G1, P[4]G2, P[8]G3, and P[8]G4) commonly seen worldwide accounted for 75.6% of the total. Taken together with uncommon strains, including the novel types P[4]G4 and P[8]G2, it highlights the extraordinary diversity of rotaviruses circulating in China. CONCLUSION: Rotavirus is the major cause of severe child diarrhea in Changchun. Developing a rotavirus vaccine for prevention of severe disease and reduction of treatment costs seemed to be necessary.

Child, Preschool↗

Comparative formation of lithocholic acid from chenodeoxycholic and ursodeoxycholic acids in the colon.

The comparative rate of formation of lithocholic acid from chenodeoxycholic acid and its 7 beta epimer, ursodeoxycholic acid, was studied in human subjects and in a rhesus monkey. [24-14C]Chenodeoxycholic acid and [24-14C]ursodeoxycholic acid were incubated in vitro, under anaerobic conditions, in fecal samples from 7 control and 7 asymptomatic gallstone subjects. The incubations were carried out for 0, 0.5, 1, 4, and 12 h. In addition, the labeled precursors were instilled into the colon of 4 asymptomatic gallstone patients and a rhesus monkey in which a bile duct fistula had been created. Radioactive metabolites were analyzed by thin-layer chromatography in the in vitro fecal incubates and in the in vivo colonic aspirates, stool, and bile. The biotransformation of the unlabeled material was analyzed by gas-liquid chromatography in the in vitro incubates and in the in vivo fecal samples of the rhesus monkey. There was no statistical difference between chenodeoxycholic and ursodeoxycholic acids in their rate of biotransformation to lithocholic acid, when the total group of 14 subjects was compared. However, among these 14, a subgroup of 4 subjects (2 controls and 2 with gallstones) was identified in whom the rate of degradation to lithocholic acid was significantly faster for chenodeoxycholic than for ursodeoxycholic acid. Increases in the concentrations of the precursors led to a decrease in the rate, but not a change in the comparative pattern of lithocholic acid formation. At the lower concentrations, the conversion of both bile acids to lithocholic acid was almost complete after 12 h. In the in vivo studies, the formation of lithocholic acid from chenodeoxycholic and ursodeoxycholic acids was comparable both in the 4 human subjects and in the rhesus monkey. The results of this study indicate that, in most cases, the risk of liver damage from lithocholic acid formation should be similar for both epimers. However, there appears to be a small population in which this risk could be higher during chenodeoxycholic acid than during ursodeoxycholic acid treatment due to a more rapid formation of lithocholic acid.

Adult↗

Levels of 234U, 238U and 230Th in excreta of uranium mill crushermen.

Pacific Northwest Laboratory conducted a research program to measure uranium and thorium levels in excreta of uranium mill crushermen who are routinely exposed to airborne uranium ore dust. The purpose of this work was to determine whether 230Th was preferentially retained in the body over either 234U or 238U. Urine and fecal samples were obtained from 14 active crushermen with long histories of exposure to uranium ore dust, plus four retired crushermen and three control individuals for comparison. Radiochemical procedures were used to separate out the uranium and thorium fractions which were then electroplated on stainless steel discs and assayed by alpha spectrometry. Significantly greater activity levels of 234U and 238U were measured in both urine and fecal samples obtained from uranium mill crushermen, indicating that uranium in the inhaled ore dust was cleared from the body with a shorter biological half-time than the daughter product 230Th. The measurements also indicated that uranium and thorium separate in vivo and have distinctly different metabolic pathways and transfer rates in the body.

Alpha Particles↗

[Acute intestinal infections of virus etiology in adults].

AIM: To describe clinical symptoms of sporadic cases of noro- and astroviral infection in adults; to compare a clinical picture and a course of rotaviral infection (R VI), noroviral infection (NVI) and astroviral infection (A VI). MATERIAL AND METHODS: Reverse transcription and polymerase chain reaction (RT-PCR) method was used to examine fecal samples from 1032 patients with acute intestinal infections (All) at the age of 15 to 90 years admitted to the infectious department of the clinic of Chelyabinsk State Medical Academy in 2004-2005. Detectability and severity of some symptoms in R VI, NVI and A VI patients were analysed and clinical manifestations of viral intestinal monoinfections were compared. RESULTS: Monoinfection was diagnosed in 230 (22.3 %)fecal samples: RVI (n = 101), NVI (n = 100), A VI (n = 29). Sporadic morbidity was the highest from March to May, March to September, October to December for R VI, NVI and A VI, respectively. Gastroenteritis syndrome was prevalent in all the infections studied. Diarrhea with marked dehydration was more typical for RVI, dyspepsia and abdominal pain--for NVI. Intoxication occurred with the same frequency in all the infections. Severe RVI ran with normal body temperature in many patients over 60 years of age. CONCLUSION: Viral diarrheal diseases are frequent in adults of all ages and run with similar clinical picture. The most serious course of the infection with most severe syndromes of gastroenteritis and dehydration was typical for RVI. Etiological nature of viral intestinal infections can be established only with laboratory diagnostic methods.

Adolescent↗

[Enzyme-linked immunosorbent assay (ELISA) in rotavirus detection].

Reagents were obtained by means of which the method of the enzyme-linked immunosorbent assay (ELISA) was worked out, based on the double sandwich principle, for the demonstration of rotavirus antigen presence in the fecal samples. The reaction makes use of polyvinylchloride plaques that are employed in tablet packing, and enables the rapid quantitative diagnosis of rotaviral enteritis in both animals and humans. The specificity of ELISA has been demonstrated through the comparative investigation of various viruses and bacterial antigens as well as by means of the periodic study of experimentally infected calves. The investigation of a total of 207 fecal samples from diseased calves showed that 28.5 per cent reacted positively for rotaviruses by the immunodiffusion test, and 42 per cent--by ELISA. Rotavirus enterites showed an expressed age-associated curve from the first to the fifteenth day of age with a peak between the fourth and the seventh day.

Animals↗

Control of Isospora suis-induced coccidiosis on a swine farm.

Results of a program designed to control neonatal porcine coccidiosis on a total confinement, farrow-to-finish swine farm are reported. The control program consisted of washing, phenol disinfection, and steam cleaning of farrowing houses and treatment of sows with amprolium HCl before and after farrowing. Before initiation of the control program, 88.9% of the sows examined in the farrowing house were negative for coccidian oocysts, 9.9% were positive for Eimeria spp, and 1.2% were positive for Isospora suis. Most pigs nursing on sows before initiation of the control program had diarrhea at 5 to 10 days of age, which led to dehydration and weight loss. Morbidity was high, and mortality was moderate. Composite fecal samples from these litters were all positive (100%) for I suis. After initiation of the control program, 99.6% of the sows examined in the farrowing house were negative for coccidian oocysts and 0.4% were positive for Eimeria spp. Clinical signs of coccidiosis were rarely present in nursing pigs examined after the control program was initiated; however, I suis was still present in 19.8% of the composite fecal samples from pigs examined. An association between oocyst production in sows and I suis infections in pigs was not found in the present study. Oocysts of Eimeria spp were not found in the feces from the pigs.

Amprolium↗

Optimization of terminal restriction fragment polymorphism (TRFLP) analysis of human gut microbiota.

Some compounds originating from the human gut microbial metabolism of exogenous and endogenous substrates may have properties that profoundly affect the host's physiological processes. The influence of these metabolites on differences in disease risk among individuals could be mediated by metabolism specific to the gut microbial community composition. In this study, we evaluated the effectiveness of terminal restriction fragment polymorphism (TRFLP) as a biomarker of the fecal microbial community (as a surrogate of gut microbiota) for application in human population-based studies. We tested the effects of experimental conditions on DNA quality, DNA quantity, and TRFLP patterns derived from gut bacterial communities. Genomic DNA was extracted from fecal slurries and the bacterial 16S rDNA genes were amplified and analyzed by TRFLP. We found that the composition of the TRFLP fingerprints varied by different extraction procedure. The best quality and quantity of community DNA extracted from fecal material was obtained by using the QIAamp DNA stool minikit (Qiagen, Valencia, CA) with 95 degrees C incubation and moderate bead beating treatment during the cell-lysis step. Homogenization of fecal samples reduced variation among replicates. Once the TRFLP procedure was optimized, we assessed the methodological and inter-individual variation in gut microbial community fingerprints. The methodological variation ranged from 4.5-8.1% and inter-individual variation was 50.3% for common peaks. In conclusion, standardized TRFLP is a robust, reproducible, and high-throughput method that will provide a useful biomarker for characterizing gut microbiota in human fecal samples.

Adult↗

Possible antibiotic-associated colitis in a dog.

A Poodle referred for renal disease developed severe colonic disease characterized by total mucosal collapse and necrosis. The onset of colonic disease was temporally related to administration of antibiotics. On 3 occasions, bacterial culture of fecal samples yielded only Streptococcus pyogenes, Pseudomonas aeruginosa, and Candida albicans, suggesting that the colonic bacterial flora was severely disrupted. Findings, although not conclusive, were suggestive of antibiotic-associated colitis that ultimately proved fatal. Colonoscopy should be considered for dogs with unduly severe large-bowel diarrhea associated with antibiotic treatment and can be done with minimal restraint and bowel preparation, if necessary. Nonselective bacterial culture of fecal samples should be considered for dogs with unduly severe large-bowel diarrhea associated with antibiotic treatment.

Ampicillin↗

Campylobacter fetus diarrhea in a Hutterite colony: epidemiological observations and typing of the causative organism.

Following a case of Campylobacter fetus sepsis and meningitis in a 4-month-old female member of a Hutterite colony, an epidemiological investigation revealed at least 18 cases of diarrhea in other members of the colony. C. fetus was isolated from 7 of 15 fecal samples submitted from affected persons. A case control study suggested that persons who worked in the abattoir were 2.03 times more likely to have had diarrhea, but none of the risk factors studied were significant. The epicurve of the outbreak was inconclusive as to the likely mode of spread of C. fetus. All of the C. fetus strains isolated from the blood of the infant and from the fecal samples were the same by biochemical and antibiotic susceptibility tests. Pulsed-field gel electrophoresis showed that all isolates produced identical restriction endonuclease patterns and differed from other nonepidemiologically related strains of C. fetus.

Adult↗

Survival of Clostridium difficile and its toxins in equine feces: implications for diagnostic test selection and interpretation.

Although Clostridium difficile is recognized as a cause of enterocolitis in horses and humans, there has been little work published regarding the lability of C. difficile and its toxins in feces. A significant decrease in recovery of C. difficile from inoculated equine fecal samples occurred during storage. Recovery after storage in air at 4 degrees C decreased from 76% (37/49) after 24 hours to 67% (33/49) at 48 hours and 29% (14/ 49) after 72 hours. In contrast to aerobic storage, 25 of 26 samples stored anaerobically at 4 degrees C yielded growth of C. difficile for 30 days, whereas the organism was only detected for 2.5 +/- 2.52 days (x +/- SD) in paired samples stored aerobically. The use of an anaerobic transport medium was effective in maintaining viability of C. difficile. These findings indicate that poor aerotolerance is the reason for the rapid decrease in culture yield. In contrast to C. difficile organisms stored aerobically at 4 degrees C, C. difficile toxins were considerably more stable and could be detected by enzyme-linked immunosorbent assay in both broth and inoculated fecal samples for at least 30 days. The poor survival of C. difficile but the stability of its toxins when feces are stored aerobically must be considered when submitting samples for diagnosis of C. difficile-associated enterocolitis in horses and when interpreting laboratory results.

Animals↗