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PubMed2005

[Prevalence of intestinal helminths in dogs from Quindío Province].

BACKGROUND: Intestinal helminths are pathogens for domestic animals and provide a source of potential infection for humans. OBJECTIVES: The prevalence of intestinal helminths in domestic dogs was determined in a province-wide survey in Quindío Province, Colombia. MATERIALS AND METHODS: The sample size was calculated based upon the data of the 2003 antirabies vaccination program in Quindio. Information in the form of an epidemiological questionnaire was provided by dog owners. Fecal samples from dogs were analyzed by Ritchie's concentration method. RESULTS: Of 324 samples, 67.6% were from purebred dog races and 32.4% were mongrels. A 22.2% prevalence for intestinal helminthes was found. Ancylostoma caninum was the most prevalent parasite (13.9%), followed by Trichuris vulpis (4.3%), Toxocara canis (2.5%), and Strongyloides stercoralis (4.0%). Multiparasite infestations were observed in 2.46% of the dogs. CONCLUSION: Presence of parasites was strongly correlated with age and degree of association with the open streets. Control programs are recommended for helminth surveillance in human and canine populations.

Animals↗
PubMedDate not supplied

[Cyclosporosis: an emergent parasitosis. (II) Microbiological diagnosis through a new staining technique].

PURPOSE: Describe a new staining technique for the diagnosis of Cyclospora infection through lab tests. A differential microscopic diagnosis between Cyclospora and Blastocystis hominis is conducted. MATERIALS AND METHODS: The research was conducted by using fecal samples referred to a private lab in the city of Lima, Peru. A direct fresh assay was made by using a saline solution and Lugol's solution. Very thin fecal smear were prepared, and a staining technique using gentian violet was applied. RESULTS: Microphotography of Cyclospora cysts are shown with the fresh assay method and the new staining technique. Moreover the morphological and structural differences between Cyclospora and Blastocystis hominis were shown: the latter is larger, shows poor staining, and has vacuoles. CONCLUSIONS: The routine use of this technique is recommended for all fecal tests due to its simplicity and low cost, and because it allows a quick diagnosis of this type of infection that has a high prevalence in our country so it may be treated prematurely and specifically.

Animals↗
PubMed1991

Detection of cryptosporidium oocysts in acute diarrheal stools.

Three hundred and fifty stool samples from patients with acute diarrhea and 30 samples from normal healthy controls were examined for the presence of various parasitic and bacterial enteropathogens by conventional techniques. Cryptosporidium oocysts were detected in 15 out of 350 (4.3%) fecal samples. In 10 cases (2.9%) it was identified as a sole pathogen while in 5 cases (1.4%) it was isolated in association with cysts of E. histolytica, Giardia lamblia and with Salmonella typhimurium. Ten out of fifteen cases were in the pediatric age group. None of the 30 control cases had Cryptosporidium oocysts in their stools.

Acute Disease↗
PubMed1981

Mycobacterium paratuberculosis in the semen and genital organs of a semen-donor bull.

Mycobacterium paratuberculosis, the cause of Johne's disease, was isolated from the feces of a donor bull in an artificial insemination stud. During isolation and observation for 21 months, the organism was recovered from all of 26 fecal samples and from 8 of 31 semen samples. At necropsy, it was isolated from the intestine and adjacent lymph nodes, lung, spleen, seminal vesicles, and prostate gland but not from the testicles. We concluded that routine fecal cultures at bull studs will reveal infected bulls before they become genitally infected and shed M paratuberculosis in semen.

Animals↗
PubMed2000

Development of a selective medium for isolation of Helicobacter pylori from cattle and beef samples.

Helicobacter pylori has been isolated from the human stomach with media containing only minimal selective agents. However, current research on the transmission and sources of infection requires more selective media due to the higher numbers of contaminants in environmental, oral, and fecal samples. The objective of this study was to develop and evaluate detection techniques that are sufficiently selective to isolate H. pylori from potential animal and food sources. Since H. pylori survives in the acidic environment of the stomach, low pH with added urea was studied as a potential selective combination. H. pylori grew fairly well on H. pylori Special Peptone plating medium supplemented with 10 mM urea at pH 4. 5, but this pH did not sufficiently inhibit the growth of contaminants. Various antibiotic combinations were then compared, and a combination consisting of 10 mg of vancomycin per liter, 5 mg of amphotericin B per liter, 10 mg of cefsulodin per liter, 62,000 IU of polymyxin B sulfate per liter, 40 mg of trimethoprim per liter, and 20 mg of sulfamethoxazole per liter proved to be highly selective but still allowed robust colonies of H. pylori to grow. This medium was highly selective for recovering H. pylori from cattle and beef samples, and it is possible that it could be used to enhance the recovery of this bacterium from human and environmental samples, which may be contaminated with large numbers of competing microorganisms.

Abomasum↗
PubMed1997

PCR detection, identification to species level, and fingerprinting of Campylobacter jejuni and Campylobacter coli direct from diarrheic samples.

Three sets of primers were designed for PCR detection and differentiation of Campylobacter jejuni and Campylobacter coli. The first PCR assay was designed to coidentify C. jejuni and C. coli based on their 16S rRNA gene sequences. The second PCR assay, based on the hippuricase gene sequence, identified all tested reference strains of C. jejuni and also strains of that species which lack detectable hippuricase activity. The third PCR assay, based on the sequence of a cloned (putative) aspartokinase gene and the downstream open reading frame, identified all tested reference strains of C. coli. The assays will find immediate application in the rapid identification to species level of isolates. The assays combine with a protocol for purification of total DNA from fecal samples to allow reproducible PCR identification of campylobacters directly from stools. Of 20 clinical samples from which campylobacters had been cultured, we detected C. jejuni in 17, C. coli in 2, and coinfection of C. jejuni and Campylobacter hyointestinalis in 1. These results were concordant with culture and phenotypic identification to species level. Strain typing by PCR-restriction fragment length polymorphism of the flagellin (flaA) gene detected identical flaA types in fecal DNA and the corresponding campylobacter isolate. Twenty-five Campylobacter-negative stool samples gave no reaction with the PCR assays. These PCR assays can rapidly define the occurrence, species incidence, and flaA genotypes of enteropathogenic campylobacters.

Amidohydrolases↗
PubMed2006

Reactions of chlorogenic acid and quercetin with a soy protein isolate--influence on the in vivo food protein quality in rats.

Plant phenolic compounds are known to interact with proteins producing changes in the food (e. g., biological value (BV), color, taste). Therefore, the in vivo relevance, especially, of covalent phenol-protein reactions on protein quality was studied in a rat bioassay. The rats were fed protein derivatives at a 10% protein level. Soy proteins were derivatized with chlorogenic acid and quercetin (derivatization levels: 0.056 and 0.28 mmol phenolic compound/gram protein). Analysis of nitrogen in diets, urine, and fecal samples as well as the distribution of amino acids were determined. Depending on the degree of derivatization, the rats fed with soy-protein derivatives showed an increased excretion of fecal and urinary nitrogen. As a result, true nitrogen digestibility, BV, and net protein utilization were adversely affected. Protein digestibility corrected amino acid score was decreased for lysine, tryptophan, and sulfur containing amino acids.

Amino Acids↗
PubMed2004

Real-time PCR of host DNA in feces to study differential exfoliation of colonocytes between rats and humans.

BACKGROUND: Colonic mucosa has a high turnover rate. At the end of their lifespan, colonocytes become senescent and die. Histological studies indicate that senescent colonocytes are shed (exfoliated) into the fecal stream in rats, but phagocytosed by mucosal macrophages in humans. We study whether quantification of host DNA in feces can be used as a non-invasive marker for this differential disposal of colonocytes. METHODS: Selective primers and probes for the rat and human beta-globin genes were designed and used in real-time PCR reactions. RESULTS: Host DNA was quantitatively extracted and detected in fecal samples of both species. Feces of rats fed a humanized diet contained approximately 100 microg rat DNA per g freeze-dried feces. In human feces, however, only 5 out of 12 samples contained detectable, though very low (less than 0.35 microg/g), levels of host DNA. This about 300-fold difference could not be attributed to differences in DNase activities in the fecal stream. CONCLUSION: Our results indicate that there is considerable luminal shedding of senescent colonocytes in rats, whereas mucosal phagocytosis is the main route of colonocyte disposal in humans. Thus, real-time PCR of host DNA in feces can be applied as a non-invasive method for studying the differential exfoliation of colonocytes.

Animals↗
PubMed2006

Clonal dissemination of Escherichia coli O157:H7 subtypes among dairy farms in northeast Ohio.

To ascertain the extent to which indistinguishable strains of Escherichia coli O157:H7 are shared between farms, molecular characterization was performed on E. coli O157:H7 isolates recovered during a longitudinal study of 20 dairy farms in northeast Ohio. Of the 20 dairy farms sampled, 16 were located in a primary area and 4 were located in two other distant geographical areas. A total of 92 E. coli O157:H7 isolates obtained from bovine fecal samples, water trough sediment samples, free-stall bedding, and wild-bird excreta samples were characterized. Fifty genetic subtypes were observed among the isolates using XbaI and BlnI restriction endonucleases. Most restriction endonuclease digestion profiles (REDPs) were spatially and temporally clustered. However, four REDPs from multiple sources were found to be indistinguishable by pulsed-field gel electrophoresis between four pairs of farms. The geographical distance between farms which shared an indistinguishable E. coli O157:H7 REDP ranged from 9 to 50 km, and the on-farm sources sharing indistinguishable REDPs included cattle and wild bird feces and free-stall bedding. Within the study population, E. coli O157:H7 REDP subtypes were disseminated with considerable frequency among farms in close geographic proximity, and nonbovine sources may contribute to the transmission of this organism between farms.

Animal Husbandry↗
PubMed1988

Detection of respiratory syncytial virus and rotavirus by enhanced chemiluminescence enzyme-linked immunosorbent assay.

The horseradish peroxidase-catalyzed oxidation of luminol was used in a chemiluminescence avidin-biotin enzyme-linked immunosorbent assay (CL-ABE) to detect respiratory syncytial virus (RSV) and rotavirus (ROV). This CL-ABE was carried out in a new apparatus constructed for the photographic registration of the light emission produced. When measured in a spectrophotometer, the light emission produced by the oxidation of luminol showed a peak emission at 425 nm. The chemiluminescence output reached maximum within 1 min after the initiation of luminol oxidation and diminished to one-half of maximum emission within 8 min. When titrations of purified ROV by avidin-biotin enzyme-linked immunosorbent assay (ABE) were monitored by CL-ABE and by conventional orthophenylenediamine (OPD) staining (OPD-ABE), the detection limits were 0.01 and 0.04 ng of ROV protein, respectively. Similar titrations of purified RSV gave detection limits of 0.2 and 0.8 ng of RSV protein by CL-ABE and OPD-ABE, respectively. When 26 RSV-positive samples of nasopharyngeal secretions (NPS) were titrated by CL-ABE and by OPD-ABE, the mean titers obtained were 737 and 254, respectively. When 19 ROV-positive fecal samples were titrated by CL-ABE and by OPD-ABE, the mean titers obtained were 82,000 and 29,000, respectively. When samples of NPS from 123 infants and children with acute lower respiratory disease were tested for the presence of RSV, 31 NPS samples were RSV positive by CL-ABE, and 29 of these 31 NPS samples were RSV positive by OPD-ABE.

Child, Preschool↗
PubMed2006

[Epidemiological importance of humans and domestic animals as reservoirs of verocytotoxin-producing Escherichia coli].

BACKGROUND/AIM: A "new" pathogenic agent, verocytotoxin--producing Escherichia coli (VTEC) emerged in the last 20 years, causing an increased number of sporadic cases, as well as of outbreaks of diarrhoeal diseases. Humans and animals can be infected with VTEC, but their epidemiological importance as a reservoir of this agent is not quite clear, especially in the Balkan region. Therefore, the aim of this study was to investigate the frequency of isolation of VTEC from the intestinal tract of humans and animals and to determine the serogroups of the isolated strains. METHODS: A total of, 3 401 stool samples from humans and 2 660 samples from five different species of domestic animals were tested for the presence of this pathogen. RESULTS: VTEC was isolated from 20 (0.6%) humans stools and from 431 (16.2%) animal fecal samples (p < 0.001). Only 15 (3.3%) VTEC strains belonged to human infection-associated serogroups (O26, O55, O111, O128 and O157), designated as enterohaemorrhagic E. coli (EHEC). The most known serogroup-O157 was identified in 6 (1.3%) of the isolated VTEC strains; of them, 1 (5%) was of human origin and 5 (1.2%) were animal strains. CONCLUSION: This study revealed that domestic animals were a more important reservoir of VTEC than humans, and that the isolated VTEC strains rarely belonged to O157, as well as to other EHEC serogroups that might explain rare sporadic cases and the absence of epidemic occurrence of diarrhoeal diseases caused by VTEC in this geographic region.

Animals↗
PubMed2001

An epidemiological investigation of congenital Schistosoma japonicum transmission in Hubei Province, PR China.

A cohort study was conducted in Hubei Province, China, following serious flooding of the Yangtze River in the autumn of 1998 to investigate the possibility of congenital transmission of Schistosoma japonicum in humans. The cohort investigated was comprised of 205 women and their 208 infants born between 1 September and 30 December 1998. Blood and fecal samples from all the women and their infants were collected and examined for S. japonicum infection. Positive specific antibody titers were found in 14 (6.8%) of the mothers, but no fecal egg excretion was observed. All infants had negative specific antibody titers and no S. japonicum eggs were found in their feces. Hence, the present study coud not confirm congenital S. japonicum transmission in humans. Further studies are highly wanted to study the impact of prenatal exposure of S. japonicum on the offspring.

Animals↗
PubMedDate not supplied

Immunogenicity of a killed E. coli 0111:K58 vaccine administered orally to children.

A killed E. coli 0111:K58 vaccine was administered by the oral route to 16 children ranging from 3 months to 7 years of age. The antibody response was measured at different times after the vaccination through the titration of agglutinating antibodies in serum and in fecal extracts, and the detection of specific IgA immunoglobulins in the intestinal content. All vaccines developed antibody response detectable both inserum and in fecal samples. Coproantibodies tended to appear earlier and to attain higher titers than circulating antibodies; they were all least in part IgA immunoglobulins, since the content of specific IgA in the fecal extracts was proportional to the agglutinating titer.

Administration, Oral↗
PubMedDate not supplied

[Epidemiologic study of an outbreak of echovirus type-9 meningitis].

BACKGROUND: Several agents are able to produce lymphocytic meningitis, but sometimes it's not possible their identification. The viruses are the etiological agents more frequently found, especially enteroviruses, mumps virus and herpes simplex virus, with different epidemiological patterns depending on time and geographic location. Most of the infections caused by enteroviruses are asymptomatics. In general the viral meningitis have a good prognostic with an acute benign course and serious signs of neurological affectation are infrequent. From 1991 it has been observed an increase of nonpolio enteroviral meningitis outbreaks in our Country. Echovirus-4 was isolated in most of the outbreaks notified during that year. Echovirus-9 was not isolated in any of them. METHODS: We describe the epidemiological and clinical characteristics of a lymphocytic meningitis outbreak that took place from June to July of 1993 in Burgos. RESULTS: Forty-eight patients, most of them children, were hospitalized with fever, headache, vomits and stiff neck with an increase in the total cell count in cerebospinal fluid (CSF). Echovirus-9 was isolated from fecal samples in eight patients. CONCLUSIONS: The etiology was attributed to Echovirus-9 because of microbiologic and epidemiologic findings. The incubation period can fluctuate between four an five days and fecal-oral transmission is the most probable mechanism.

Adolescent↗
PubMed1993

[Endoparasites--prevention in ungulates in fenced enclosures: experiences with a pilot project in the Dählhölzli Zoo in Bern].

In the animal zoo Dählhölzli Berne all ungulates are continuously monitored since 1984 (qualitatively and quantitatively) in the context of an epidemiological study on gastrointestinal strongyles. Fecal sampling occurs every two weeks in the "parasite risk group", every four to six weeks in the so called "non-risk group". Risk group-animals repeatedly show increased fecal egg counts, whereas among non-risk group-animals only ascarid eggs in Przewalski-horses and wild boars can become significant. The animals are treated (metaphylactically) only at a high egg production or at a steadily increasing egg production. A resulting slight but constant reinfection allows the build-up of an immunity, leading thereby to a decreased frequency of treatment. During the monitoring period (1984-1990) only four animals out of 80 post-mortem analysed animals had a diagnoses of an endoparasitoses as a cause of death.

Animals↗
PubMed2006

Survival of Escherichia coli O157:H7 in bovine feces over time under various temperature conditions.

Although Escherichia coli O157:H7 prevalence estimates in cattle have increased over time due to improvements in detection methods, fecal sample transport conditions from farm to microbiological laboratories for further analysis may be a factor for prevalence underestimation. The objective of this study was to compare and determine the survival characteristics of E. coli O157:H7 in bovine feces under various storage conditions that could be encountered during transport. Fecal pats were inoculated with a four-strain cocktail of antibiotic-resistant E. coli O157:H7 to contain approximately 1 x 10(5) CFU/g. Inoculated and control samples were taken after 0, 24, 48, 120, and 168 h at each storage temperature and examined for presence and numbers of E. coli O157:H7. Each sample was subdivided and placed at each of the four following temperatures: 37, 23, 4.4 degrees C, and in a plastic cooler with refrigerant packs (0, 4, 4, 21, and 23 degrees C at five sampling times, respectively) to simulate transportation conditions. A statistically significant decrease in the population of the pathogen was observed after 48 h in samples held at 37 degrees C (P < 0.01) and after 168 h at 4.4 degrees C (P = 0.02). At 37 degrees C, E. coli O157:H7 was not detected after 48 h, either by direct plating (P < 0.01) or by immunomagnetic separation. Overall, the results of this study showed that E. coli O157:H7 survived without significant detriment in bovine fecal material inside the cooler for up to 168 h. These results indicate that shipment and storage under these conditions before microbiological analysis would be acceptable and should not affect pathogen detection.

Animals↗
PubMed1997

Isolation and virulence factors of verocytotoxin-producing Escherichia coli in human stool samples.

OBJECTIVE: To evaluate the isolation rate of O157 and non-O157 verocytotoxin-producing Escherichia coli (VTEC) strains, to study the occurrence of additional virulence factors and to correlate these with clinical symptoms. METHODS: Over more than 5 years, 17 296 unduplicated fecal samples submitted for routine culture were screened for VTEC by a single PCR detecting VT1, VT2 and its variants. Verocytotoxin B subunit genotypes of the isolates obtained by testing individual colonies in positive samples were determined by a polymerase chain reaction---restriction fragment length polymorphism (PCR---RFLP) technique, the eaeA gene and the 60-MDa virulence plasmid by PCR, and the hemolytic phenotype by using CaCl2-washed blood agar. RESULTS: Verocytotoxin genes were found in 1.02% of the samples. Non-O157 VTEC strains were isolated in 0.66% and O157 in 0.17%. Overall, VTEC was less frequently isolated than Campylobacter and Salmonella but more frequently than Yersinia and Shigella. All cases except two siblings were epidemiologically unrelated. Cases of hemolytic uremic syndrome (HUS) were only observed in association with serogroup O157, which seems to be more pathogenic than the non-O157 strains. Among non-O157 VTEC strains, eaeA-positive strains are more frequently associated with clinical symptoms than are eaeA-negative strains. Other virulence factors correlate less closely with the presence of symptoms. CONCLUSIONS: VTEC is the third bacterial intestinal pathogen in our study population. All stool samples from patients with diarrhea should be screened for the most frequent serogroup, O157, or, if this is not possible, at least those from patients with bloody diarrhea. Non-O157 VTEC strains, especially if they are eaeA positive, are also associated with diarrhea, more often non-bloody. PCR or the new commercially available immunoassays could be used in selected cases, e.g. in patients suffering from HUS and in cases of outbreaks.

Journal Article↗
PubMed2006

Blood lead concentrations in waterfowl utilizing Lake Coeur d'Alene, Idaho.

The Coeur d'Alene River Basin, Lake Coeur d'Alene, and the Spokane River contain elevated heavy metal concentrations in sediment and water from historical mining and ore processing operations in the Coeur d'Alene Basin. Lead poisoning has been identified as the cause of death in hundreds of waterfowl utilizing wetlands in the floodplain of the Coeur d'Alene River, but little was known about hazards to waterfowl from heavy metal contamination in shallow bays and wetlands of Lake Coeur d'Alene. We examined lake sediment and blood lead concentrations in waterfowl utilizing Lake Coeur d'Alene, Idaho, to evaluate potential lead contamination of waterfowl utilizing the lake. We collected 56 palustrine and 102 lacustrine sediment samples and 61 mallard and 8 wood duck blood samples. Mean lead concentrations from palustrine and lacustrine sediment samples ranged from 14 to 3508 mg/kg dry weight (dw) and from 19 to 5009 mg/kg (dw), respectively. Lead concentrations in palustrine and lacustrine sediment from several Lake Coeur d'Alene bays were higher than those in lake reference areas and were higher than Bunker Hill Superfund Site target cleanup levels and suggested site-specific toxicity thresholds for swans. Mean blood lead from mallard and wood ducks sampled from Lake Coeur d'Alene bays were within lead toxicity ranges for waterfowl associated with clinical and severe clinical lead poisoning. We also collected 19 Canada goose and 3 mallard fecal samples to evaluate exposure through sediment ingestion. Waterfowl using Lake Coeur d'Alene appear to be exposed to lead by ingesting contaminated lake sediment. Our model predicts a sediment lead effects range of 147-944 mg/kg (dw) and mortality effects level of 1652 mg/kg (dw) for mallards utilizing Lake Coeur d'Alene. The locations of Harrison Slough, Powderhorn Bay, and Cottonwood Bay at the mouth of the Coeur d'Alene River and Blackwell Island and Cougar Bay near the Spokane River outflow of Lake Coeur d'Alene were the areas of greatest concern for waterfowl exposure to lead contaminated sediment.

Animals↗