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Seasonal variation in the endocrine-testicular function of captive jaguars (Panthera onca).

Captive adult male jaguars (Panthera onca) from two locations in southeast Brazil were studied to evaluate the effects of season on endocrine and testicular function. For assessment of testicular steroidogenic activity, androgen metabolite concentrations were measured in fecal samples collected one to three times per week over 14 ( n=14 ), 9 ( n=1 ) or 7 months ( n=1 ). To assess seasonality, data were grouped by season (summer: December-February; autumn: March-May; winter: June-August; spring: September-November). Additionally, samples collected in the dry season (March-August) were compared with those collected in the wet season (September-February). There were no differences ( P>0.05 ) in fecal androgen concentrations in samples collected in spring, summer, autumn, and winter ( 480.8+/-50.4 ng/g, 486.4+/-42.0 ng/g, 335.4+/-37.7 ng/g, and 418.6+/-40.4 ng/g dry feces). However, there were differences ( P<0.05 ) in fecal androgen concentrations between the dry and wet seasons ( 380.5+/-28.0 ng/g versus 483.9+/-32.3 ng/g dry feces). Sperm samples, collected from all males twice (approximately 6 months apart) were similar; mean (+/-S.E.M.) motility, concentration and morphology were 57.0 %4.5%, 6.3+/-2.4 x 10(6) ml(-1), and 60.8+/-3.1 %, respectively. In conclusion, androgen metabolite concentrations in the captive male jaguar were not affected by season, but there was a difference between the wet and dry periods. Further research is needed to verify these results.

Androgens↗

Effect of inclusion of fermentable carbohydrates in the diet on fermentation end-product profile in feces of weanling piglets.

An in vivo experiment was conducted to monitor the changes in fermentation end products in the feces of weaning piglets due to the inclusion of selected fermentable carbohydrates in the diet. The experiment involved 3 groups of 16 piglets each. Specially raised piglets (neither antibiotics nor creep feeding) were weaned abruptly at 4 wk of age. The piglets were offered 1 of 2 dietary treatments [a control diet (CON), or a fermentable carbohydrate-enriched diet (CHO)] and were subjected to 1 of the 2 fasting treatments (fasting for 2 d at the beginning of the experimental period or nonfasting). Fecal samples were collected per rectum every day during the experimental period. Piglets were slaughtered at the end of the 10-d experimental period, and digesta samples were collected from different parts of the gastrointestinal tract (GIT): the first half of the small intestine, the second half of the small intestine, the cecum, and colon. The DM, VFA profile, and ammonia concentrations were analyzed from the fecal and digesta samples. Daily feed intake was also recorded. There was no difference in concentrations of VFA in feces between the treatment groups. Ammonia concentration was lower (P < 0.05) in piglets fed the CHO diet compared with those fed the CON diet in both feces and digesta from different parts of GIT. Fasting had no effect on fermentation end products in feces. This study demonstrated that the inclusion of fermentable carbohydrates in weanling diets reduces protein fermentation along the GIT and also reduced the fecal concentration of ammonia.

Acetic Acid↗

Evaluation of deuterated cholesterol and deuterated sitostanol for measurement of cholesterol absorption in humans.

The continuous isotope feeding method of Crouse and Grundy (1978. J. Lipid Res. 19: 967-971) for measurement of dietary cholesterol absorption has been modified by using markers labeled with stable isotopes ([2,2,4,4,6-2H5]cholesterol or [25,26,26,26,27,27,27-2H4]cholesterol or [26,26,26,27,27,27-2H6] cholesterol and [5,6,22,23-2H4]sitostanol) quantified by gas-liquid chromatography-selected ion monitoring. Tracing of the isotope distribution of the authentic markers and after their intestinal passage, including the microbiological products (coprostanol and coprostanone) revealed stability of the labels. The new method was evaluated in six monkeys on two occasions by comparison with the original method using radioactively labeled cholesterol and sitosterol. The results obtained by the two different methods were in excellent agreement, and absorption ranged from 49% to 73% (mean 60%) for the stable isotope method and from 51% to 69% (mean 62%) for the radioactive method. The coefficient of variation of cholesterol absorption in animals ranged from 3.9% to 15.1% (mean 7.1%) for stable isotopes and 1.9% to 13.6% (mean 5.7%) for radioactive isotopes. In twelve subjects cholesterol absorption was measured by the new method from total fecal samples frozen immediately and compared to results obtained from small fecal aliquots (approximately 1 g) sent by ordinary mail to the laboratory. A significant correlation of cholesterol absorption between the two different sample handlings was obtained (r = 0.981, P < 0.001). In addition, measurement of cholesterol absorption twice in seven volunteers 2 weeks apart revealed identical results. Thus, the new method is extremely safe and reproducible without radioactive exposure to the subjects and labortory staff and can be used on women of child-bearing age.

Animals↗

Urine as another potential source for template DNA in polymerase chain reaction (PCR).

This technical note examines the potential for preparing template DNA in polymerase chain reactions (PCR) from urine in Japanese macaques (Macaca fuscata). Microsatellite band patterns from urine samples showed close agreement with those of blood and fecal samples, and only a few hundred microl of urine yielded a template DNA for PCR. This research will increase the opportunity for scientists to examine the genetic backgrounds of their target animals by using non-invasive sample collection in the wild.

Animals↗

Genome-targeted enrichment and sequencing of human-infecting Cryptosporidium spp.

Cryptosporidium spp. are parasites that cause severe illness in vulnerable human populations. Obtaining pure and sufficient Cryptosporidium DNA from clinical and environmental samples is a challenging task. Oocysts shed in available fecal samples can be limited in quantity, require purification (biased towards dominant strains), and yield limited DNA (&#x2009;<&#x2009;40 fg/oocyst). Here, we use updated genomic sequences from a broad diversity of Cryptosporidium species that have been found to infect humans (C. cuniculus, C. hominis, C. meleagridis, C. parvum, C. tyzzeri, and C. viatorum) to develop and validate a set of 100,000 RNA baits (CryptoCap_100k) with the aim of enriching Cryptosporidium DNA from varied samples. Compared to unenriched libraries, CryptoCap_100k increases the percentage of reads mapping to target genome sequences, increases the depth and breadth of genome coverage, and facilitates analyses of genetic variants in many samples, while decreasing overall costs.

Cryptosporidium↗

Oral administration of arabinogalactan affects immune status and fecal microbial populations in dogs.

Seven ileally cannulated dogs were randomly assigned to a control or arabinogalactan (AG) treatments in a 7 x 7 Latin square design to evaluate effects of oral AG administration on nutritional and immunological characteristics. Arabinogalactan treatments included a high (1.65 g/d) and low (0.55 g/d) dose of AG100, AG1000 or AG3000 provided via gelatin capsules. Arabinogalactan forms differed in purification procedures. Each period consisted of a 6-d adaptation followed by a 4-d collection. Blood and fresh fecal samples were collected on d 10 of each period. Fecal score increased (P < 0.02) in dogs supplemented with the low dose of AG1000. Ileal and total tract dry matter (DM) and organic matter (OM) digestibilities were not affected by treatment. Dogs supplemented with the high dose of AG1000 tended (P = 0.15) to have a higher concentration of total aerobic fecal bacteria than control dogs. Dogs supplemented with the low dose of AG1000 and the high dose of AG3000 had higher concentrations of fecal lactobacilli (P = 0.04) and tended to have higher concentrations of fecal bifidobacteria (P < or =0.16) compared with control dogs. Dogs fed the low dose of AG3000 tended (P = 0.10) to have a lower concentration of fecal Clostridium perfringens compared with control dogs. Arabinogalactan treatments did not affect (P > 0.05) serum immunoglobulin G, M or A concentrations. Specific forms and doses of AG increased white blood cell, neutropil and eosinophil concentrations. Arabinogalactan is a unique dietary fiber that affects the digestive physiology and immunological characteristics of dogs.

Animals↗

GENOME TARGETED ENRICHMENT AND SEQUENCING OF HUMAN-INFECTING CRYPTOSPORIDIUM spp.

Cryptosporidium spp. are parasites that cause severe illness in vulnerable human populations. Obtaining pure and sufficient Cryptosporidium DNA from clinical and environmental samples is a challenging task. Oocysts shed in available fecal samples can be limited in quantity, require purification (biased towards dominant strains), and yield limited DNA (<40 fg/oocyst). Here, we use updated genomic sequences from a broad diversity of human-infecting Cryptosporidium species ( C. cuniculus , C. hominis , C. meleagridis , C. parvum , C. tyzzeri , and C. viatorum ) to develop and validate a set of 100,000 RNA baits (CryptoCap_100k) with the aim of enriching Cryptosporidium spp. DNA from varied samples. Compared to unenriched libraries, CryptoCap_100k increases the percentage of reads mapping to target genome sequences, increases the depth and breadth of genome coverage and the reliability of detecting species and mixed infections within a sample, and allows assessment of genetic variation via SNP calling, while decreasing costs.

Journal Article↗

Rapid and effective method for preparation of fecal specimens for PCR assays.

We have developed a novel method for the preparation of fecal specimens for PCR assays. Approximately 100 mg of solid stool or 200 microliters of liquid fecal sample was thoroughly suspended in 1 ml of water. Fecal debris was removed by low-speed centrifugation (2,800 x g for 2 min). The supernatant was then boiled for 10 min in a water bath and further clarified by high-speed centrifugation (12,000 x g for 5 min). Fifty microliters of the clarified supernatant was then purified by Sepharose CL-6B spin column chromatography, and a portion of the purified supernatant was used for PCR. By this method, stools containing enterotoxigenic Escherichia coli H10407 were amplified by colonization factor antigen I fimbrial gene PCR, with a sensitivity of 100 organisms per reaction. The method was also effective for processing stool specimens for Clostridium difficile toxin A and B gene PCRs. This method is rapid, effective, and simple to perform and will improve the applications of PCR to stool specimens for diagnostic purposes.

Bacterial Typing Techniques↗

Detection and differentiation of bovine group A rotavirus serotypes using polymerase chain reaction-generated probes to the VP7 gene.

Dot and Northern blot hybridization assays were developed to detect and differentiate group A bovine rotavirus serotypes using radiolabeled serotype 6 (Nebraska calf diarrhea virus [NCDV] and United Kingdom [UK] strains) or serotype 10 (Crocker [Cr] strain) VP7 gene probes. Partial length VP7-specific cDNA encompassing areas of major sequence diversity were generated by the polymerase chain reaction (PCR) using either cloned VP7 genes (NCDV and UK strains) or reverse transcribed mRNA (Cr strain) as templates. Radiolabeled probes prepared from the PCR-generated cDNA were tested at various stringency conditions to optimize the hybridization assays. At high stringency conditions (52 C, 50% formamide, 5 x standard saline citrate), the NCDV, UK, and Cr probes serotypically differentiated bovine rotavirus isolates in RNA samples prepared from cell culture propagated viruses or in fecal specimens from infected gnotobiotic calves. The sensitivity and specificity of NCDV and Cr VP7 probes were characterized in dot blot hybridization assays, and the probes were estimated to detect at least 1 ng of viral RNA. The serotyping results obtained using VP7 probes were similar to those obtained using serologic assays. Further development of these assays may provide a useful means for the rapid detection and differentiation of bovine rotavirus serotypes in fecal samples from calves in the field.

Animals↗

Metabolic and microbial responses in western crossbred and Meishan growing pigs fed a high-fiber diet.

Four Duroc x White composite crossbred (21.8 +/- 1.0 kg BW) and four 12-wk-old Meishan purebred (20.7 +/- 1.6 kg BW) growing barrows were used to determine the relative breed differences in metabolic and microbial responses to a high-fiber diet. The pigs were trained to consume 700 g of a diet containing 35% (as-fed basis) dehydrated alfalfa meal once daily. The pigs' daily intakes of DM, N, GE, NDF, and ADF were 610 g, 16.6 g, 2.64 Mcal, 150 g, and 88 g, respectively. On d 12 after surgical catheterization of the portal vein, ileal vein, and carotid artery, a 3-d total urine and feces collection was conducted. On d 24 after surgery, each pig was placed in an open-circuit calorimeter, and its catheters were connected to a system for simultaneous measurements of oxygen consumption by portal-drained viscera and by whole body, and the net portal absorption of VFA after a 24-h fasting and during a 5-h postprandial period. The VFA measured included acetic, propionic, isobutyric, butyric, isovaleric, and valeric acids. A second 3-d total urine and feces collection was conducted on d 30 after surgery. There were no differences (P = 0.13) between the first and second collections in apparent total-tract digestibility coefficients for nutrients and N retention of pigs. Compared with Duroc x White composite crossbred pigs, Meishan pigs had lower (P = 0.05) apparent digestibility coefficients for DM, N, NDF, hemicellulose, and N retention, but their portal-drained viscera used a greater (P = 0.05) fraction of whole-body oxygen consumption. No differences (P = 0.12) were found between Duroc x White composite crossbred and Meishan pigs in total viable bacteria and cellulolytic bacteria from fecal samples, in vitro digestibility of alfalfa NDF fractions by fecal inocula, whole-body oxygen consumption, net portal absorption of VFA, total energy of absorbed VFA, and the potential of absorbed VFA for meeting the energy needs for whole-body heat production. These results indicate that, in contrast to previous beliefs, the ability of Meishan growing pigs to utilize a high-fiber diet is not superior to that of Duroc x White composite crossbred growing pigs.

Animal Feed↗

Impact of commercial pre-harvest processes on the prevalence of Salmonella enterica in cull sows.

This study examined the impact of transport and lairage on Salmonella enterica prevalence in cull sows held under commercial conditions. Five sampling periods over 10 weeks yielded 181 selected culls. Fecal sample positives at the farm and collection point were 2% and 3%, respectively. After transport and lairage 41% of cull sows yielded S. enterica in one or more sampled tissues. A significant increase in cecal isolations was observed. The two S. enterica serotypes found at the production site and collection point were found at the abattoir, as were 13 other serotypes. Transport and lairage practices may contribute to an increased S. enterica contamination immediately prior to slaughter.

Abattoirs↗

A retrospective study of lead poisoning in cattle.

Information pertaining to 294 cases of bovine lead poisoning that occurred during the 15-year period from 1968 to 1982 was collected from the toxicology laboratory records at the Western College of Veterinary Medicine. Lead poisoning was seen more often in younger cattle, with 51.9% of the cases reported in animals 6-months of age or less. A statistically significant seasonal incidence was seen (p less than 0.0001) with 60.9% of the cases occurring from May to August. Lead poisoning was more common in dairy breeds than beef breeds. The relative rates in the daily and beef breeds per 1,000 cattle were 0.022 and 0.0065 respectively. No specific breed susceptibility was associated with any of the major dairy or beef breeds. Statistically significant sex-related differences for lead poisoning were not seen (p = 0.11). The most suitable biological samples utilized to confirm lead poisoning were blood, kidney and liver. These samples produced the most reliable results, with rumen and fecal samples exhibiting a high degree of variability.

Age Factors↗

Cryptosporidium spp. in ruminants at the Lisbon Zoo.

Feces from 34 species of ruminants housed at the Lisbon Zoo was examined for Cryptosporidium spp. oocysts. Three hundred and eighty-eight samples were analyzed. Three hundred and eighty species-specific group fecal samples were collected monthly, from September 1998 until August 1999, along with eight individual specimens from eight neonates. All samples were examined by four different techniques: microscopic observation of direct and concentrated fecal smears, staining with modified Ziehl-Nielsen, immunofluorescent assay, and immunoenzymatic assay. The prevalence of infection was 3.6%. Five neonates with diarrhea were infected. Cryptosporidial oocysts were shed more frequently during winter months. Some facilities may have permitted oocysts to remain viable, possibly contributing to cryptosporidial transmission between animals.

Animals↗

[Fecal calprotectin in differential diagnosis of irritable bowel syndrome].

OBJECTIVE: To assess the value of fecal calprotectin as a non-invasive screening biomarker in differential diagnosis of irritable bowel syndrome compared with fecal occult blood test (FOBT), erythrocyte sedimentation (ESR) or C reactive protein (CRP). METHODS: Subjects were a total of 240 persons, including 60 patients with irritable bowel syndrome, 60 patients with colorectal cancer, 60 patients with chronic inflammation, and 60 healthy controls. 5 g fecal samples were collected within one week of endoscopy or before surgical operation. Fecal calprotectin was measured by an enzyme-linked immunosorbent assay (ELISA) kit in spot stool samples. At the same time, FOBT was measured; the results of ESR and CRP in hospital lab were collected. RESULTS: The median of fecal calprotectin concentrations were 12.21 mg/kg and 15.36 mg/kg in IBS and healthy controls, respectively. There was no statistical significance of calprotectin concentration between patients with IBS and healthy controls (P>0.05). The median of fecal calprotectin concentrations were 159.00 mg/kg and 466.00 mg/kg in colorectal cancer and chronic inflammation respectively. There were statistical significance between patients with chronic inflammation, colorectal cancer, and others (P<0.01). The maximal calprotctin concentration was with chronic inflammation; the medium with colorectal cancer; the minimum with IBS and healthy controls. When the cut-off limit was set as 50 mg/kg of fecal calprotectin, the positive rates of colorectal cancer, chronic inflammation, IBS and healthy controls were 85.0%, 91.7%, 10%, and 5%,respectively. Fecal calprotectin was much superior to FOBT, ESR and CRP. CONCLUSION: Fecal calprotectin as a non-invasive screening biomarker in the differential diagnosis of IBS and symptomatic chronic large intestinal organic disease was better than FOBT, ESR and CRP. It was simple, inexpensive, repeatable and no-invasive. It can be used as a biomarker in exclusion from related organic diseases before the diagnosis of irritable bowel syndrome.

Adolescent↗

Correlation of enterohemorrhagic Escherichia coli O157 prevalence in feces, hides, and carcasses of beef cattle during processing.

A survey was performed to estimate the frequency of enterohemorrhagic Escherichia coli O157:H7 or O157:nonmotile (EHEC O157) in feces and on hides within groups of fed cattle from single sources (lots) presented for slaughter at meat processing plants in the Midwestern United States, as well as frequency of carcass contamination during processing from cattle within the same lots. Of 29 lots sampled, 72% had at least one EHEC O157-positive fecal sample and 38% had positive hide samples. Overall, EHEC O157 prevalence in feces and on hides was 28% (91 of 327) and 11% (38 of 355), respectively. Carcass samples were taken at three points during processing: preevisceration, postevisceration before antimicrobial intervention, and postprocessing after carcasses entered the cooler. Of 30 lots sampled, 87% had at least one EHEC O157-positive preevisceration sample, 57% of lots were positive postevisceration, and 17% had positive postprocessing samples. Prevalence of EHEC O157 in the three postprocessing samples was 43% (148 of 341), 18% (59 of 332) and 2% (6 of 330), respectively. Reduction in carcass prevalence from preevisceration to postprocessing suggests that sanitary procedures were effective within the processing plants. Fecal and hide prevalence were significantly correlated with carcass contamination (P = 0.001), indicating a role for control of EHEC O157 in live cattle.

Animals↗

Relationship among fecal coliforms, Escherichia coli, and Salmonella spp. in shellfish.

The relationship of fecal coliforms, Escherichia coli, and Salmonella spp. was examined in freshly harvested and stored shellfish. In 16 of 40 freshly collected oyster samples, fecal coliform levels were above the recommended wholesale level suggested by the National Shellfish Sanitation Program (less than or equal to 230/100 g), and Salmonella spp. were present in three of these samples. Salmonella spp. were not, however, present in any sample containing less than 230 fecal coliforms per 100 g. Analysis of the data suggests that low fecal coliform levels in both fresh and stored oysters are good indicators of the absence of Salmonella spp., but that high levels of fecal coliforms are somewhat limited in predicting the presence of Salmonella spp. E. coli levels correlated very strongly with fecal coliform levels in both fresh and stored oysters and clams, suggesting that there is no advantage in replacing fecal coliforms with E. coli as an indicator of shellfish quality.

Animals↗

Screening adenoviruses in stool samples: evaluation of a genus-specific monoclonal antibody based enzyme immunoassay.

To screen fecal samples for adenovirus antigens a genus-specific monoclonal antibody based enzyme immunoassay was developed. In a comparative analysis with commercial latex agglutination test, high sensitivity was demonstrated. The assay did not detect other viruses usually found in faeces suggesting its specificity. One hundred and eighty stool samples collected in Baranya County were tested and 13 (7.2%) of them showed reactivity. The application of our immunoassay combined with other, more sophisticated methods may help us to determine the serotype specificity of these adenovirus isolates and assess the importance of adenoviruses in viral gastroenteritis.

Adenovirus Infections, Human↗