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PubMed1996

Efficacy of moxidectin 0.5% pour-on against naturally acquired nematode infections in cattle.

The efficacy of 0.5% moxidectin pour-on at two dosages was evaluated in 30 cattle with naturally acquired gastrointestinal nematode infections. The cattle were ranked according to pretreatment fecal egg counts and body weights, and were randomly assigned from replicates to three treatment groups. The mean number of trichostrongyle eggs in pretreatment fecal samples did not differ among the groups. Groups I and II received 0.5% moxidectin pour-on at dosages of 0.25 mg moxidectin per kg body weight and 0.5 mg moxidectin per kg body weight, respectively, and Group III cattle received moxidectin-free vehicle (control cattle). The cattle were euthanized by complete replicate 14 or 15 days posttreatment for recovery of nematodes and examination of the pour-on sites. Treatment with moxidectin was associated with a significant reduction in fecal trichostrongyle egg counts compared with the control cattle; efficacy was 99.7% and > 99.9% for cattle in Groups I and II, respectively. Eleven species of adult nematodes (Haemonchus contortus, Haemonchus placei, Ostertagia ostertagi, Trichostrongylus axei, Trichostrongylus colubriformis, Bunostomum phlebotomum, Cooperia oncophora, Cooperia punctata, Nematodirus helvetianus, Oesophagostomum radiatum and Trichuris ovis) and 4th stage larvae of Ostertagia spp. and Trichostrongylus spp. were present in sufficient numbers in control cattle to evaluate the efficacy of moxidectin 0.5% pour-on. In all cases, treatment was associated with a significant reduction in the number of nematodes recovered at necropsy, with the efficacy of both dosages exceeding 99.9%. There were no significant differences in mean worm burdens or fecal egg counts between the two treated groups, and no adverse reactions were observed.

Abomasum↗
PubMed1987

Dietary fiber supplementation and fecal bile acids, neutral steroids and divalent cations in rats.

Young adult rats were fed ad libitum for 4 wk on defined diets containing either no fiber, 10% levels of insoluble fiber sources [cellulose, wheat bran, alfalfa, mixed fibers (Fibyrax)], 5% levels of soluble fiber sources [pectin, guar gum, psyllium (Metamucil)] or 2% cholestyramine (Questran), a bile acid sequestrant. Fecal samples were obtained from paired rats over a 3-d period, were analyzed for neutral and acidic steroid levels and compositions and, together with the food, were assayed for divalent cations (Ca2+, Mg2+, Fe2+ and Zn2+). Animals in all groups were in balance for divalent cations, and there appeared to be no relationship between the extent of cation balance and the type of fiber fed. All insoluble fiber diets and the guar gum and psyllium diets resulted in significantly higher daily fecal mass and, in general, resulted in significant dilution of total fecal steroids. With the insoluble fibers, there was a general dilution of fecal neutral steroids, which was not apparent with the soluble fibers or cholestyramine. In addition, except for the pectin- and mixed fiber-containing diets, there was reduced bacterial conversion of the primary bile acids to secondary bile acids or metabolites.

Animals↗
PubMed1989

Identification of Clostridium difficile in stool specimens by culture-enhanced gas-liquid chromatography.

We have developed a sensitive and specific method for the identification of Clostridium difficile in stool specimens based on the detection of metabolic breakdown products of the organism by gas-liquid chromatography after incubation of stool samples in a selective broth medium containing cefoxitin. Use of this approach to test samples from two different populations of patients at separate medical centers showed this method to be superior to plate cultures or cytotoxin testing alone for both populations. The combined results from the two patient populations showed that 225 of 226 confirmed isolates were identified correctly, resulting in a sensitivity of 99.6% and a specificity of 99.0%. This method eliminates the delay caused by subculturing for tests requiring a pure isolate. The culture phase amplifies even low numbers of C. difficile in fecal samples (due to low in vivo concentrations or delayed transport) and thus increases sensitivity. Other advantages include the ability to detect C. difficile in the mixed flora of the stool and the ability of most clinical laboratories to use this procedure. Given the complexities of the detection of C. difficile toxins and the increasing importance of this organism as a nosocomial agent, culture-based methods remain the preferred approach to screening and routine workup for cases of diarrhea.

Adult↗
PubMed2005

Production and characterization of monoclonal antibodies against excretory/secretory products of adult Echinococcus granulosus, and their application to coproantigen detection.

Two IgM murine monoclonal antibodies (MAbs), EgC1 and EgC3, were produced against the excretory/secretory (E/S) products of Echinococcus granulosus adult worms. Immunoblotting revealed that both predominantly recognized a 50 kDa antigen in the somatic extract and an 85 kDa component in the E/S products. Immunolocalization showed that both MAbs reacted with the tegument of the parasite, and additionally EgC3 reacted with parenchyma and the tegument lining the external surface of the reproductive organs. A coproantigen capture ELISA was developed using a rabbit polyclonal antibody against E/S products from adult tapeworms as catching antibodies, and each one of MAbs as detecting antibody. The assays detected seven out of eight (EgC1), and eight out of eight (EgC3) experimentally infected dogs (worm burdens ranging from 61 to 57,500), using heat-treated samples obtained at prepatent period, and none (n=8) of helminth-free samples. Time course analysis showed that, after a 12-25 days lag, coproantigen levels rose above cut off O.D. values and typically peaked around 30 days post-infection (DPI) at the end of the experiment. One dog experimentally infected with Taenia hydatigena metacestodes was slightly detected as positive at different time points after 30 DPI. Both MAbs showed a similar pattern of recognition, but T. hydatigena antigens were undetectable for a longer period, and reached lower O.D. values with EgC1. Interestingly, fecal samples from two experimentally infected dogs with Echinococcus multilocularis were not recognized by the EgC1 assay, suggesting a potential value as species-specific diagnostic tool.

Animals↗
PubMed2004

Effects of subcutaneous injections of a long acting moxidectin formulation in grazing beef cattle on parasite fecal egg reduction and animal weight gain.

Trials were conducted in Arkansas, Idaho, Illinois and Wisconsin using a common protocol to evaluate effectiveness and safety of a long acting (LA), oil-based injectable formulation of moxidectin in beef cattle grazing spring and/or summer pastures. At each site, 150 cattle (steers and/or heifers) were blocked based on pretreatment fecal strongyle egg counts (EPG) and then randomly assigned to treatments within blocks. Presence of naturally acquired parasitic infections, confirmed by presence of parasite eggs in feces, was a prerequisite for study enrollment. Within each block of three animals, two received moxidectin LA injectable on day 0 at a dosing rate of 1.0 mg moxidectin/kg b.w. into the dorsal aspect of the proximal third of the ear, and one received a placebo control treatment. Cattle were weighed before treatment and on day 55 or 56 (55/56) after treatment. Fecal samples were also collected from 10 randomly selected blocks of animals at each site on days 14, 28 and 55/56 for EPG quantification. Average daily gain (ADG) was computed over the posttreatment period. Data pertaining to ADG and EPG were combined across sites and analyzed by mixed model analysis of variance to assess the fixed effect of treatment and random effects of site, block within site and the treatment by site interaction. Compared to placebo-treated controls, the geometric means of fecal EPG counts from cattle treated with moxidectin LA injectable were reduced 99.8% 14 days after treatment, 99.1% 28 days after treatment and 96.7% 55/56 days after treatment. Rate of weight gain by cattle treated with moxidectin LA injectable was 0.59 kg/day, or 23% (0.11 kg/day) more than placebo-treated controls (P<0.05). None of the cattle treated with moxidectin LA injectable exhibited signs of macrocyclic lactone toxicosis. Summarized across all study sites, proportions of cattle that received concurrent therapeutic treatments were similar among treatment groups. Study results demonstrate that moxidectin cattle LA injectable administered at a dosing rate of 1.0 mg moxidectin/kg b.w. to grazing beef cattle was effective and safe.

Analysis of Variance↗
PubMed2002

Molecular diversity of Lactobacillus spp. and other lactic acid bacteria in the human intestine as determined by specific amplification of 16S ribosomal DNA.

A Lactobacillus group-specific PCR primer, S-G-Lab-0677-a-A-17, was developed to selectively amplify 16S ribosomal DNA (rDNA) from lactobacilli and related lactic acid bacteria, including members of the genera Leuconostoc, Pediococcus, and WEISSELLA: Amplicons generated by PCR from a variety of gastrointestinal (GI) tract samples, including those originating from feces and cecum, resulted predominantly in Lactobacillus-like sequences, of which ca. 28% were most similar to the 16S rDNA of Lactobacillus ruminis. Moreover, four sequences of Leuconostoc species were retrieved that, so far, have only been detected in environments other than the GI tract, such as fermented food products. The validity of the primer was further demonstrated by using Lactobacillus-specific PCR and denaturing gradient gel electrophoresis (DGGE) of the 16S rDNA amplicons of fecal and cecal origin from different age groups. The stability of the GI-tract bacterial community in different age groups over various time periods was studied. The Lactobacillus community in three adults over a 2-year period showed variation in composition and stability depending on the individual, while successional change of the Lactobacillus community was observed during the first 5 months of an infant's life. Furthermore, the specific PCR and DGGE approach was tested to study the retention in fecal samples of a Lactobacillus strain administered during a clinical trial. In conclusion, the combination of specific PCR and DGGE analysis of 16S rDNA amplicons allows the diversity of important groups of bacteria that are present in low numbers in specific ecosystems to be characterized, such as the lactobacilli in the human GI tract.

Adult↗
PubMed2003

Measurement of plasma corticosterone and fecal glucocorticoid metabolites in the chicken (Gallus domesticus), the great cormorant (Phalacrocorax carbo), and the goshawk (Accipiter gentilis).

A method for the non-invasive measurement of glucocorticoid metabolites in feces of chickens was established and validated. After high-performance liquid chromatography (HPLC) the presence of at least two fecal immunoreactives was demonstrated, one co-eluting with authentic corticosterone, whereas the second substance migrates close to corticosterone sulphate. We investigated the relationship between corticosterone in blood plasma obtained by a vena brachialis catheter and fecal samples in groups of five chickens after an ACTH and a dexamethasone injection to stimulate and to suppress adrenal activity. A control group received a saline injection. After ACTH plasma cortisol concentrations increased 16-fold after 1.5 h to levels between 19 and 38 ng/ml and dropped to pre-treatment levels (1.1-2.5 ng/ml) 4h after stimulation. Dexamethasone did not result in a distinct suppression of adrenocortical activity and plasma corticosterone dropped only slightly below pre-treatment levels. The concentrations in fecal metabolites corresponded to the changes in the levels of biological active hormone in plasma. Fecal peak excretion (105-295 ng/g) was obtained with a delay of approximately 4 h compared to plasma. The profile obtained after ACTH challenge reflected a broader and dampened pattern of glucocorticoid secretion and provided a more integrated measure of adrenal activity. Dexamethasone treatment did not induce a measurable decrease in fecal metabolites and concentrations fluctuated around a mean of 30.0+/-9.9 ng/g, almost identical to those obtained from the saline treatment group (29.4+/-13.9 ng/g). In a separate experiment the effect of an alternative capture method (remote-controlled injection system) was investigated in cormorants. Plasma corticosterone measurements revealed a significantly diminished stress reaction compared to traditional trapping (1.24+/-0.78 vs. 10.9+/-12.1 ng/ml). Investigations whether goshawk nestlings infected with Trichomas gallinae differ in fecal corticosterone metabolite concentrations compared to healthy subjects revealed no significant changes. However, a significant correlation was found between the glucocorticoid metabolite concentrations and the number of nestlings per nest. The demonstration that adrenal activity can be detected by the assay is a prerequisite that ecologically meaningful levels of imposed stress can be validated. Therefore, non-invasive measurements of fecal metabolites are a promising perspective to monitor stress in birds.

Adrenocorticotropic Hormone↗
PubMed2003

Fecal alpha1-proteinase inhibitor concentration in dogs with chronic gastrointestinal disease.

BACKGROUND: Fecal alpha(1)-proteinase inhibitor (alpha(1)-PI) clearance is a reliable, noninvasive marker for protein-losing enteropathy in human beings. An assay for use in dogs has been developed and validated. OBJECTIVE: The aim of this study was to evaluate fecal alpha(1)-PI concentration in dogs with chronic gastrointestinal disease, compared with healthy dogs, and to assess its correlation with serum albumin concentration. METHODS: Fecal samples were collected from 2 groups of dogs. Group 1 consisted of 21 clinically healthy client-owned dogs without signs of gastrointestinal disease. Group 2 consisted of 16 dogs referred for investigation of suspected gastrointestinal disease. On the basis of gastric and duodenal biopsies, group 2 was further subdivided into dogs with normal histology (n = 9) and those with histologic abnormalities (n = 7: inflammatory bowel disease, n = 3; lymphangiectasia, n = 4). An ELISA was used to measure alpha(1)-PI concentrations in fecal extracts. RESULTS: Fecal alpha(1)-PI concentrations, expressed as micro g/g of feces, were not significantly different between groups 1 and 2 as a whole. However, fecal alpha(1)-PI concentrations (median, minimum-maximum) were significantly higher in dogs with gastrointestinal diseases associated with histologic abnormalities (60.6 micro g/g, 7.4-201.7 micro g/g) compared with dogs with normal histology (3.8 micro g/g, 0.7-74.0 micro g/g) and control dogs (9.9 micro g/g, 0.0-32.1 micro g/g). There was no significant correlation between fecal alpha(1)-PI and serum albumin concentrations in dogs with gastrointestinal disease. CONCLUSIONS: Increased fecal alpha(1)-PI concentration may signal the need to obtain gastrointestinal biopsies for a final diagnosis. Fecal alpha(1)-PI concentration may be a useful test for early detection of protein-losing enteropathy before decreases in serum albumin concentration can be detected.

Animals↗
PubMedDate not supplied

Disposition of 14C-eptifibatide after intravenous administration to healthy men.

Eptifibatide, a synthetic peptide inhibitor of the platelet glycoprotein IIb/IIIa receptor, has been studied as an antithrombotic agent in a variety of acute ischemic coronary syndromes. The purpose of the present study was to characterize the disposition of 14C-eptifibatide in man after a single intravenous (i.v.) bolus dose. 14C-Eptifibatide (approximately 50 microCi) was administered to eight healthy men as a single 135-microgram/kg i.v. bolus. Blood, breath carbon dioxide, urine, and fecal samples were collected for up to 72 hours postdose and analyzed for radioactivity by liquid scintillation spectrometry. Plasma and urine samples were also assayed by liquid chromatography with mass spectrometry for eptifibatide and deamidated eptifibatide (DE). Mean (+/- SD) peak plasma eptifibatide concentrations of 879 +/- 251 ng/mL were achieved at the first sampling time (5 minutes), and concentrations then generally declined biexponentially, with a mean distribution half-life of 5 +/- 2.5 minutes and a mean terminal elimination half-life of 1.13 +/- 0.17 hours. Plasma eptifibatide concentrations and radioactivity declined in parallel, with most of the radioactivity (82.4%) attributed to eptifibatide. A total of approximately 73% of administered radioactivity was recovered in the 72-hour period following 14C-eptifibatide dosing. The primary route of elimination was urinary (98% of the total recovered radioactivity), whereas fecal (1.5%) and breath (0.8%) excretion was small. Eptifibatide is cleared by both renal and nonrenal mechanisms, with renal clearance accounting for approximately 40% of total body clearance. Within the first 24 hours, the drug is primarily excreted in the urine as unmodified eptifibatide (34%), DE (19%), and more polar metabolites (13%).

Adult↗
PubMed2006

Effects of Lactobacillus casei Shirota, Bifidobacterium breve, and oligofructose-enriched inulin on colonic nitrogen-protein metabolism in healthy humans.

Pre- and/or probiotics can cause changes in the ecological balance of intestinal microbiota and hence influence microbial metabolic activities. In the present study, the influence of oligofructose-enriched inulin (OF-IN), Lactobacillus casei Shirota, and Bifidobacterium breve Yakult on the colonic fate of NH3 and p-cresol was investigated. A randomized, placebo-controlled, crossover study was performed in 20 healthy volunteers to evaluate the influence of short- and long-term administration of OF-IN, L. casei Shirota, B. breve Yakult, and the synbiotic L. casei Shirota + OF-IN. The lactose[15N,15N]ureide biomarker was used to study the colonic fate of NH3. Urine and fecal samples were analyzed for 15N content by combustion-isotope ratio mass spectrometery and for p-cresol content by gas chromatography-mass spectrometry. RT-PCR was applied to determine the levels of total bifidobacteria. Both short- and long-term administration of OF-IN resulted in significantly decreased urinary p-cresol and 15N content. The reduction of urinary 15N excretion after short-term OF-IN intake was accompanied by a significant increase in the 15N content of the fecal bacterial fraction. However, this effect was not observed after long-term OF-IN intake. In addition, RT-PCR results indicated a significant increase in total fecal bifidobacteria after long-term OF-IN intake. Long-term L. casei Shirota and B. breve Yakult intake showed a tendency to decrease urinary 15N excretion, whereas a significant decrease was noted in p-cresol excretion. In conclusion, dietary addition of OF-IN, L. casei Shirota, and B. breve Yakult results in a favorable effect on colonic NH3 and p-cresol metabolism, which, in the case of OF-IN, was accompanied by an increase in total fecal bifidobacteria.

Adult↗
PubMed1981

Thiamin status of a Melanesian population.

A semiautomated method is described which uses the Abbott ABA-100 bichromatic analyzer to measure the stimulation of erythrocyte transketolase by thiamin pyrophosphate (the thiamin pyrophosphate effect). The method was accurate with a throughput of 30 samples per hour and is suitable for the study of the thiamin status of large populations. A significant proportion (21%) of a total of 1920 individuals of a predominantly Melanesian population were found to have an elevated thiamin pyrophosphate effect (greater than 15% activation). This same proportion with an elevated thiamin pyrophosphate effect was found in an urban hospital patient group of 346 individuals. Statistical analysis indicated a significant inverse correlation between thiamin pyrophosphate effect and plasma potassium concentration (p less than 0.001). Analysis of human fecal samples for thiaminase I activity demonstrated that this enzyme was present in 23 of a total of 46 samples selected from both the Melanesian and urban hospital patient groups. Preliminary findings indicated that the thiaminase originated from a bacterial source.

Adolescent↗
PubMed2003

Longitudinal study of Salmonella enterica aerotype Typhimurium infection in three Danish farrow-to-finish swine herds.

A longitudinal study of the infection dynamics of Salmonella enterica was carried out with three Danish farrow-to-finish swine herds. To account for variations in Salmonella shedding over time, litters from each herd were divided into two cohorts. Each cohort consisted of 30 pigs, for a total of 180 pigs. Pigs were individually monitored by monthly bacteriologic and serologic examinations from weaning to slaughter. At weaning, individual sows were examined bacteriologically and serologically. At slaughter, cecal contents, ileocecal lymph nodes, and carcass swab samples were obtained from 131 pigs. A total of 88 pigs were found to be shedding Salmonella on one or more occasions. Only the Salmonella serotype Typhimurium was detected during the study period. At weaning, no sows or piglets were found to be shedding, but a serological reaction was detected in 11 sows. The prevalence in culture peaked in the nursery and subsequently declined to undetectable levels before slaughter. The seroprevalence peaked approximately 60 days after the peak prevalence in culture. Salmonella was detected in individual fecal samples at least once in 53% of the pigs, and 62% of the pigs were seropositive more than once. Only 3.7% of all pigs were found to be culture positive on more than one occasion. Piglets from seroreacting sows had a significantly (P = 0.0339) lower probability of shedding in the nursery. Under the assumption that shedding lasted at least 1 or 2 weeks, the average shedding time was estimated to have been 18 or 26 days. An association between serology, on-farm bacteriology, and Salmonella prevalence in culture at slaughter was shown. Marked differences in prevalence in sera and prevalence in culture between cohorts and within herds were observed. These differences emphasize the need for caution when using point estimates in on-farm interventions and surveillance in subclinically infected swine herds.

Animals↗
PubMed2004

Effect of Maillard reaction products on ruminal and fecal acid-resistant E. coli, total coliforms, VFA profiles, and pH in steers.

Six ruminally cannulated Angus-cross steers (362 kg) were used in a replicated 3 x 3 Latin square design to determine effects of supplementing Maillard reaction products (MRP) on acid-resistant E. coli and coliform populations. Steers were fed roughage-based diets supplemented (DM basis) with either 10% soybean meal (SBM), 10% nonenzymatically browned SBM (NESBM), or 10% SBM top-dressed with 45 g of a lysine-dextrose Maillard reaction product (LD-MRP). Equal weights of dextrose, lysine hydrochloride, and deionized water were refluxed to produce the LD-MRP. The NESBM was manufactured by treating SBM with invertase enzyme, followed by heating to induce nonenzymatic browning. Steers were allowed slightly less than ad libitum access to diets fed twice daily and were adapted to their respective treatments within 10 d. On d 11, ruminal and fecal samples were collected at 0, 2, 4, 6, 8, and 12 h after feeding from each of the steers and transported to the laboratory for microbial analysis. Ruminal samples and feces were analyzed for pH and VFA, and both ruminal fluid and feces were tested for acid-resistant E. coli and total coliforms by incubating samples in tryptic soy broth adjusted to pH 2, 4, and 7. Ruminal pH and total VFA concentrations did not differ among treatments. The molar proportion of ruminal acetate was higher (P < 0.05) for steers receiving NESBM than for steers receiving SBM and LD-MRP. At pH 4, steers that received NESBM had lower (P < 0.05) ruminal populations of E. coli and total coliforms than steers that received SBM. No differences were observed for ruminal E. coli and total coliforms at pH 2 and 7. Fecal pH was lower (P < 0.05) for steers fed NESBM than for steers fed SBM or LD-MRP. Molar proportions of fecal acetate were lower (P < 0.05) and proportions of butyrate and isovalerate were higher (P < 0.05) for steers fed NESBM compared with steers fed SBM. Fecal E. coli at pH 4 was lower (P < 0.05) for steers fed NESBM than for steers fed LD-MRP. Fecal E. coli and total coliforms at pH 2 and 7 did not differ among treatments. Dietary MRP had limited effectiveness at decreasing acid-resistant coliforms in the rumen and feces of cattle. Acid resistance in coliforms may depend on protein availability.

Animal Feed↗
PubMed2006

Effects of probiotic or prebiotic supplemented milk formulas on fecal microbiota composition of infants.

The aim of the study was to evaluate whether supplementation of milk-formulas with prebiotic fructo-oligosaccharides or a probiotic, Lactobacillus johnsonii La1 (La1), could modulate the composition of the fecal microbiota of formula-fed infants, compared to breastfed (BF) infants. Ninety infants close to 4 months of age were randomized into one of three groups to be blindly assigned to receive for 13 weeks: a) an infant formula (Control), b) the same formula with fructo-oligosaccharides (Prebio), or c) with La1 (Probio). At the end of this period, all infants received the control formula for 2 additional weeks. Twenty-six infants, breastfed throughout the study, were recruited to form group BF. Fecal samples were obtained upon enrolment and after 7 and 15 weeks. Bacterial populations were assessed with classical culture techniques and fluorescent in situ hybridisation (FISH). Seventy-six infants completed the study. On enrolment, higher counts of Bifidobacterium and Lactobacillus and lower counts of enterobacteria were observed in BF compared to the formula-fed infants; these differences tended to disappear at weeks 7 and 15. No major differences for Clostridium, Bacteroides or Enterococcus were observed between the groups or along the follow up. Probio increased fecal Lactobacillus counts (p<0.001); 88% of the infants in this group excreted live La1 in their stools at week 7 but only 17% at week 15. Increased Bifidobacterium counts were observed at week 7 in the 3 formula groups, similar to BF infants. These results confirm the presence of higher counts of bifidobacteria and lactobacilli in the microbiota of BF infants compared to formula-fed infants before dietary diversification, and that La1 survives in the infant digestive tract.

Bifidobacterium↗
PubMed2002

A longitudinal study of gastrointestinal parasites in Canadian dairy farms. The value of an indirect Ostertagia ostertagi ELISA as a monitoring tool.

The general objective of this study was to evaluate a crude Ostertagia ostertagi antigen enzyme-linked immunosorbent assay (ELISA) for monitoring gastrointestinal parasites in lactating dairy cattle. A longitudinal study of gastrointestinal parasites in lactating dairy cows was carried out in 38 herds in four provinces of Canada (Prince Edward Island, Quebec, Ontario and Saskatchewan) from September 1999 to October 2000. Bulk tank milk, cow milk, serum and fecal samples were collected monthly or quarterly from all these farms. Information on herd management factors was collected by a standard questionnaire and individual cow production data were obtained from an electronic database. The overall mean optical density ratio (ODR) was 0.30 and ranged from -0.05 to 1.55. Although a clear seasonal pattern was not observed, the ODR values tended to decrease during the housing period and start increasing in the spring before the cows went out to pasture. The second and third or greater lactation cows had significantly higher ODR values compared with first lactation animals. The individual cow ODR had a very low correlation with individual squared root fecal egg counts but showed a reasonably high correlation when herd averages values were computed (r=0.73). A moderate correlation (r approximately 0.50) between the bulk tank and herd average ODR was observed. Milk yield was negatively associated with individual cow milk ODR and a quadratic effect on ODR was observed for days in milk. Twenty-eight of the herds participated in a clinical trial of eprinomectin (Ivomec Eprinex) treatment at calving. The cow level ODR values determined late in the previous lactation had a marginally significant effect (P=0.07) on treatment response, suggesting that high OD cows responded better to the anthelmintic treatment. However, because of the small sample size available in this model, more research is needed to better understand this relationship. In conclusion, the indirect ELISA using an O. ostertagi crude antigen appears useful as a technique for monitoring gastrointestinal parasite burdens in adult dairy cows and holds promise as a potential predictor of response to anthelmintic treatment.

Animal Husbandry↗
PubMed2003

Viability and infectivity of Cryptosporidium parvum oocysts detected in river water in Hokkaido, Japan.

The viability and infectivity of Cryptosporidium parvum (C. parvum) oocysts, detected in water samples collected from river water in Hokkaido, were investigated using Severe Combined Immunodeficient (SCID) mice. The water samples collected from September 27 through October 10, 2001 by filtration using Cuno cartridge filters were purified and concentrated by the discontinuous centrifugal flotation method. From 1.2 x 10 (5) liters of the raw river water, approximately 2 x 10(4) oocysts were obtained and designated as Hokkaido river water 1 isolate (HRW-1). Oocyst identification was carried out using microscopic and immunological methods. Six 8-week-old female SCID mice were each inoculated orally with 1 x 10 (3) oocysts. Infection was successfully induced, resulting in fecal oocyst shedding. Oocysts were then maintained by sub-inoculation into SCID mice every 3 months. Infectivity was evaluated by making comparisons with two known C. parvum stocks, HNJ-1 and TK-1, which were bovine genotypes detected in fecal samples from a cryptosporidiosis patient and young cattle raised in Tokachi, Hokkaido respectively. The oocyst genotypes were determined from a small subunit ribosomal RNA (SSU-rRNA) gene by polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) analysis. No significant differences were observed in the average number of oocysts per gram of feces (OPG) in any of the isolates. Our data indicates that the C. parvum oocysts detected in the sampled river water were of C. parvum genotype 2. Moreover, our data on the continued isolation, detection and identification of the C. parvum isolates is consistent with the available epidemiological data for the Tokachi area.

Animals↗
PubMed1994

Digoxin disposition in elderly humans with hypochlorhydria.

Digoxin (D3) metabolism is partially mediated by the gastrointestinal tract via acid hydrolysis of digitoxose sugar moieties and bacterial reduction of the lactone. The hypothesis that hypochlorhydria influences digoxin disposition was tested in six normochlorhydric (NC) and four hypochlorhydric (HC) subjects. D3 tablets were administered daily for 19 to 28 days, and quantitative urine and fecal samples were collected over the last 3 days (steady state). Samples were analyzed for D3 and its extractable metabolites by fluorescence-derivatization HPLC. Excretion of D3 in urine increased from 37% of the dose in NC to 46% in HC, whereas excretion of D3 in feces decreased from 29 to 14%. These changes were statistically significant (P < .05) and consistent with decreased hydrolysis of D3 by stomach acid and increased intestinal metabolism in HC. In each subject, D3 was added to anaerobic cultures of both feces and jejunal fluid. Digoxin was reduced in all but two of the fecal incubates, and was not reduced in any jejunal fluid incubates. Because dihydrodigoxin (DHD3) was found in only two hypochlorhydric subjects, in vitro measures of bacterial reduction of D3 were not predictive of in vivo excretion of reduced metabolites. Sugar-hydrolyzed, reduced metabolites were not found in any subjects. It is concluded that D3 disposition is altered by hypochlorhydria, and that an understanding of the metabolic mechanisms requires further study.

Achlorhydria↗
PubMed1996

VP4 and VP7 typing using monoclonal antibodies.

Both rotavirus outer capsid proteins, VP4 and VP7, elicit neutralizing antibodies. Neutralizing mouse monoclonal antibodies (N-MAbs) to VP7 are easily derived and have been used widely and successfully to serotype both stool-derived and culture-adapted rotaviruses by enzyme immunoassay (EIA). Generally, approximately 70% of rotaviruses in stool samples are typable by VP7 EIA, an inexpensive and practical method. Variations in antigenic regions between strains within human rotavirus serotypes 1, 2, 4, and 9 have been recorded. These have been termed monotypes because they are detected with N-MAbs. The molecular basis for monotypes has been determined by mapping mutations selected in N-MAb-resistant antigenic variants, and by sequence analysis of the gene encoding VP7 in newly recognized monotypes. Antigenic regions A, B and C in VP7 are involved. In order to detect all members of a particular VP7 serotype, it is necessary to type with a panel of N-MAbs specific for that serotype. N-MAbs to VP4 of human rotavirus are difficult to raise and few have proven suitable for VP4 serotyping by EIA. The specificity of the assay for each P type is highest when the VP7 serotype specificity of the capture antiserum is matched to the G type of the rotavirus in the test sample. The VP4 EIA gives similar typing rates to the VP7 typing EIA. N-MAbs directed to VP8, the smaller subunit of VP4 generated by proteolytic cleavage, are more likely to show serotype specificity. Some N-MAbs that select mutations in the putative fusion region of VP5, the larger subunit of VP4, show cross-reactivity with extracts of normal, uninfected MA 104 cells and with fetal bovine serum. These N-MAbs also give elevated EIA OD readings with rotavirus-positive, but previously non-reactive fecal samples which have been frozen and thawed repeatedly. Overall, VP8-reactive N-MAbs appear most suitable for VP4 typing by EIA.

Animals↗