We have recently developed a new PCR assay for the detection of H. pylori. In this study, the polymerase chain reaction (PCR) assay was used to detect H. pylori in 88 gastric biopsy, 85 saliva, and 71 fecal specimens from 88 patients. H. pylori infection was confirmed in 71 of 88 patients by culture and/or histological stain of gastric biopsies. Serum IgG antibody to H. pylori was also measured and resulted in 97% sensitivity and 94% specificity. H. pylori DNA was detected by the PCR assay in gastric biopsy specimens from all 71 patients (100% sensitivity) with proven gastric H. pylori infection but not from 17 noninfected patients (100% specificity). In saliva specimens, H. pylori DNA was identified in 57 of the 68 patients (84%) with proven gastric H. pylori infection and in three of the 17 patients without gastric H. pylori infection. However, the PCR assay was only able to detect H. pylori DNA in the feces from 15 of 61 patients (25%) with proven gastric H. pylori infection and one of the 10 patients without gastric H. pylori infection. The results show that the PCR assay is reliable for detecting the presence of H. pylori in gastric biopsy and saliva specimens. The data indicate that H. pylori exists in a higher prevalence in saliva than feces and that the fecal-oral route may be an important means of transmission of this infection in developing countries but not as significant as previously suspected in the developed countries. It is likely that the oral-oral route is more prominent.

The primary objectives of the following experiments were to: 1) determine the impact of a directed microbial (DFM) blend on digestibility and microbial populations on horses fed low (LQ) and high (HQ) quality hay, 2) evaluate the differences in diet digestibility, microbial populations, and blood metabolites due to DFM supplementation to the exercising horse, 3) assess the impact of short-chain fructooligosaccharide (scFOS) supplementation on digestibility and the fecal microbial ecosystem in senior (SR) vs. mature (MA) horses, and 4) determine the relationship between insulin dysregulation (ID) and morphometric neck measurements (MNM) in the non-obese stock-type horse. Results indicated that: 1) Supplementation with DFM tended to be beneficial in enhancing CP digestibility. Feeding CP beyond requirements may contribute to excess excretion of Escherichia coli (EC) in HQ which was supported by 16S rRNA analysis. Differences in the fecal microbial ecosystem were detected between LQ and HQ. The phylum Saccharibacteria was identified in both hay qualities even though it has been lightly reported in equine literature. 2) Supplementing DFM to moderately exercised horses tended to enhance DM and ADF digestibility but there was no evidence of a dietary modulation to the fecal bacteria isolated. 3) No differences were found between MA and SR when fed scFOSded ration balancer which indicates that scFOS may help mitigate decreases in digestibility experienced by SR. Senior horses had less diversity in their fecal bacterial population, which may help explain why SR are more prone to ailments such as colic and impaired immune function. 4) Obesity is an important genetic factor for ID however it should not be the only determining factor, as ID can occur in non-obese individuals. One individual out of 62 tested horses was identified as ID. Morphometric neck measurements (MNM) may help pinpoint horses that are at a greater risk for ID, but more research is needed to validate MNM in both obese and non-obese stock-type horses. There were correlations between glucose sampled before an oral sugar test (OST) and insulin post-OST but a fasting OST would still be recommended in assessing ID.

In order to analyze the bacterial community of river otter scat (fecal material) at the class level, river otter scat samples were collected from Grizzly Island Wildlife Area (Solano County, CA) and the Cosumnes River Preserve (Sacramento County, CA). DNA was isolated from each sample with the MOBIO PowerSoil™ DNA Isolation Kit and 16S rRNA gene sequences were amplified from each sample. After digestion with Mspl, TRFLPs were analyzed in an ABI Prism™ 310 Genetic Analyzetin triplicate and data peak information from each electropherogram was uploaded into the Phylogenetic Assignment Tool (PAT). Species belonging to the Class Bacilli were the most abundant followed by unclassified species. Two road killed river otters were necropsied to recover brain and blood tissue. DNA was isolated using the Qiagen Tissue DNeasy Kit. Samples from both otters were amplified with a singe tube nested PCR primer set for the detection of the ITS 1 region of Toxoplasma gondii. Scat samples used in the T-RFLP analysis were also tested for the presence ofT. gondii using the same nested primer set. Neither the river otter tissue samples nor any of the scat samples used in this analysis showed evidence of infeGtion with T. gondii.

A importância da espécie suína na transmissão de Campylobacter spp. assemelha-se aos demais grupos de animais que se destinam à produção de carne, incluindo aves, bovinos e ovinos. Os objetivos deste estudo foram isolar Campylobacter spp. a partir de fezes e carcaças de suínos abatidos no Estado de São Paulo; identificar as espécies de Campylobacter spp. presentes nos animais abatidos; caracterizar os isolados obtidos através do Polimorfismo do Comprimento de Fragmentos Amplificados (AFLP). Para tal, foram utilizadas 120 amostras de fezes e 120 suabes de carcaças de suínos, colhidas de quatro diferentes abatedouros do Estado de São Paulo. Das 120 amostras de fezes analisadas, 30 foram positivas para o isolamento de Campylobacter coli (25%) e duas foram positivas para isolamento de Campylobacter jejuni (1,6%). Todas as amostras analisadas de suabes de carcaça foram negativas para Campylobacter spp. As estirpes isoladas que apresentaram características bioquímicas sugestivas de Campylobacter spp. foram submetidas ao teste de susceptibilidade ao ácido nalidixico e cefalotina, destas 19,16% (23/120) apresentaram resistência ao ácido nalidixico apesar de todas as características bioquímicas indicarem se tratar de Campylobacter coli. Foram selecionadas para a análise genotípica 38 amostras isoladas, sendo 36 de C. coli e dois de C. jejuni. A análise dos isolados através do AFLP revelou a presença de 28 perfis que foram designados P1 a P28. A técnica discriminou as cepas de acordo com a espécie, porém, uma cepa previamente caracterizada como C. coli foi agrupada com isolados de C. jejuni. Não foi possível estabelecer a correlação entre os isolados e o abatedouro de origem, no entanto observa-se uma forte tendência dos isolados resistentes ao ácido nalidixico em formar grupamentos de maior similaridade. / The importance of swine species in the transmission of Campylobacter spp. resembles to the other groups of animals that are destined to the meat production, including bovine and ovine animals. The objectives of this study were to isolate Campylobacter spp. from swine feces and carcasses slaughtered in São Paulo State; to identify the Campylobacter spp. species presents in the slaughtered animals; to characterize the isolated samples by amplified fragment length polymorphism (AFLP), for this, 120 swine feces samples and the same number of carcasses swabs were collected of four different slaughterhouses in São Paulo State. From 120 feces samples, 30 (25%) were positive for Campylobacter coli isolation and two (1,6%) were positive for Campylobacter jejuni. All the analyzed carcass swabs samples were negatives for Campylobacter spp. The isolated samples that presented suggestive biochemical characteristics of Campylobacter spp. were submitted to the susceptibility test to the nalidixic acid and cefalotine, from that 19.16% (23/120) presented resistance to the nalidixic acid in spite of all the biochemical characteristics indicate that they were Campylobacter coli. They were selected for the genotypic analysis 38 isolated samples, being 36 of C. coli and two of C. jejuni. The analysis of the 38 tested samples by AFLP showed the presence of 28 profiles that had been assigned P1 to P28. The technique discriminated the samples in agreement with the species, however, one sample previously characterized as C. coli was clustered as C. jejuni. It was not possible to establish the correlation between the isolated samples and the origin slaughterhouse, however a strong tendency of the isolated samples acid nalidixic resistant to create clusters with more similarity.

Thesis (M.S.)--Massachusetts Institute of Technology, Dept. of Chemical Engineering, 1982. / MICROFICHE COPY AVAILABLE IN ARCHIVES AND SCIENCE. / Bibliography: leaves 95-96. / by Cordelia Mae Price. / M.S.

Made available in DSpace on 2015-04-22T22:14:05Z (GMT). No. of bitstreams: 1 Dissertacao Final Giane Zupellari.pdf: 1538567 bytes, checksum: 99734d4093e76fed6d05c37113c7903d (MD5) Previous issue date: 2008-08-01 / Fundação de Amparo à Pesquisa do Estado do Amazonas / Human rotavirus constitutes one of the most important causes of diarrheic disease in children less than five years in the whole world. With the objective of verifying the occurrence of genotypes of human rotavirus in children of 1 the 36 months residents in the city of Manaus in Amazon, had been analyzed during the years from 2004 to 2006, a total of 607 the feces diarrheic samples that had been collected of children attended hospital of children in the city of Manaus. This analyzes was carried through by the detention of the viral genome through the techniques of EGPA (electrophoresis in gel of polyacrylamide), and RT-PCR (reaction in chain of polymerases associated to the reaction of reversal transcriptase). Through the EGPA method can be observed a positivity of 25% of the human rotavirus samples. All of the positive samples for rotavirus had been genotyped by the method of RT-PCR with use of specific primers for genotypes G (G1, G2, G3, G4 and G9) and genotypes P (P[8], P[4], P[6], P[9] and P[10]). By the genotyping, it was detected genotype G in 30,7% of the positive samples for rotavirus, with predominance of the G2 genotypes (17%), followed of G1 (14.9%) and G9 (10.6%) and genotype P in 44,7% of the positive samples for rotavirus, with predominance of genotypes P[8] in 66,2% of samples and P[4] in 13,2% of the samples. The combinations of genotypes G and P had been responsible for 53,3% of the positive samples for rotavirus, being that P[8]G1 and P[4]G2 had been found with 12,5% and 10% consecutively, followed of P[8]G3 (5%). Other combinations as P[4]G1, P[8]G9 and P[10]G1 had been also found in less expressive numbers. It was observed that 38.3% of the positive samples for genotype G and 11.7% of the positive samples for genotype P had not presented characteristics for none of searched genotypes G and P. The number positive samples for human rotavirus, as well as for genotypes G and genotypes P had been similar in such a way in children of the masculine sex, how much in children of the feminine sex, being that these samples had predominated in children less than 18 months. The positive samples for human rotavirus, as well as for genotypes G and genotype P had been found in every month of the years of had lasted the study. The east zone of the city of Manaus was where they had been found the biggest number of positive samples for rotavirus, as well as for genotype G and genotype P. The children with positive samples for rotavirus had presented on average of 10 episodes of diarrhea for child per day with predominance for the feces of liquid aspect (86.2%), this profile also was observed in the children with positive samples for genotypes G and genotypes P. With exception of the hyperthermia, other signals and characteristic symptoms of the diarrheic disease for rotavirus as vomit, dehydration and abdominal pain, had been presents in the biggest parcel of the children with positive samples in such a way for human rotavirus as for genotype G, as for genotype P. The high number of samples that had not been tipadas with use of specific primers for genotypes G and P, suggest the circulation of uncommon genotypes in the region, however so that this situation is defined, new studies of the epidemiological molecular of rotavirus circulating in Manaus, Amazon, must be developed, enclosed specific primers also for detention of rotavirus of animal origin. / Os rotavírus humano constituem uma das mais importantes causas de doença diarréica em crianças menores de cinco anos em todo o mundo. Com o objetivo de verificar a ocorrência de genótipos de rotavírus humano em crianças de 1 a 36 meses residentes no município de Manaus no Amazonas, foram analisadas durante os anos de 2004 a 2006, um total de 607 amostras de fezes diarréica que foram coletadas de crianças atendidas em prontos socorros infantis do município de Manaus. Esta análise foi realizada pela detecção do genoma viral através das técnicas de EGPA (eletroforese em gel de poliacrilamida) e RT-PCR (reação de polimerase em cadeia com transcriptase reversa). Através do método de EGPA pode-se observar uma positividade de 25% das amostras para rotavírus humano. Todas as amostras positivas para rotavírus foram genotipadas pelo método de RT-PCR com utilização de primers específicos para os genótipos G (G1, G2, G3, G4 e G9) e genótipos P (P[8], P[4], P[6], P[9] e P[10]). Pela genotipagem foi detectado o genótipo G em 30,7% das amostras positivas para rotavírus, com predominância dos genótipos G2 (17%), seguidos de G1 (14,9%) e G9 (10,6%) e o genótipo P em 44,7% das amostras positivas para os rotavírus, com predominância dos genótipos P[8] em 66,2% das amostras e P[4] em 13,2% das amostras. As combinações dos genótipos G e P foram responsáveis por 53,3% das amostras positivas para rotavírus, sendo que P[8]G1 e P[4]G2 foram as mais encontradas com 12,5% e 10% consecutivamente, seguidas de P[8]G3 (5%). Outras combinações como P[4]G1, P[8]G9 e P[10]G1 também foram encontradas em números menos expressivos. Foi observado que 38,3% das amostras positivas para o genótipo G e 11,7% das amostras positivas para o genótipo P não apresentaram características para nenhum dos genótipos G e P pesquisados. O número amostras positivas para rotavírus humano, bem como para os genótipos G e para os genótipos P foram semelhantes tanto em crianças do sexo masculino, quanto em crianças do sexo feminino, sendo que estas amostras predominaram em crianças menores de 18 meses. As amostras positivas para rotavírus humano, assim como para os genótipos G e para o genótipo P foram encontradas em todos os meses do estudo. A zona leste da cidade de Manaus foi onde foram encontradas o maior número de amostras positivas para rotavírus, assim como para o genótipo G e para o genótipo P. As crianças com amostras positivas para rotavírus apresentaram em média de 10 episódios de diarréia por criança por dia com predominância para os fezes de aspecto líquido (86,2%). Este perfil também foi observado nas crianças com amostras positivas para os genótipos G e para os genótipos P. Com exceção da hipertermia, outros sinais e sintomas característicos da doença diarréica por rotavírus como vômito, desidratação e dor abdominal, estiveram presentes na maior parcela das crianças com amostras positivas para os rotavírus humano, para o genótipo G e para o genótipo P. O número elevado de amostras que não foram tipadas com utilização de primers específicos para os genótipos G e P sugerem a circulação de genótipos incomuns na região, porém para que esta situação seja definida, novos estudos da epidemiologia molecular dos rotavírus circulantes em Manaus, Amazonas, devem ser desenvolvidos, incluindo inclusive primers específicos para detecção de rotavírus de origem animal.

Reduced availability of high energy-content prey (nutritional stress) is a predominant hypothesis to explain the decline of Steller sea lion (Eumetopias jubatus) populations in western Alaska from the late 1970'€™s to the late 1990'™s. Animals may physiologically respond to consuming insufficient prey by increasing stress levels and decreasing metabolic rates. It may thus be possible to identify nutritional stress by measuring concentrations of glucocorticoids (stress) and thyroid hormones (metabolism) shed in feces. However, techniques to measure thyroid hormone concentrations from Steller sea lion feces have not been developed. We quantified variation of triiodothyronine (T3) and thyroxine (T4) concentrations in Steller sea lion feces following injections of thyrotropin (TSH) into four captive animals. Glucocorticoids (GC) were also assayed to examine any relationship to stimulated thyroid hormone secretion. We found that fecal T3 peaked 48 h post-injection and increased 25-57% in three sea lions (all animals, p=0.03). Pre-injection GC increases indicated stress from isolation for baseline fecal collections, but post-injection increases could not be confirmed as a response to TSH injections or as a product of the study design. The results demonstrated that pre- and post-injection changes in fecal GC and T3 concentrations were consistent with predictions of an increased stress response and metabolic rate within the animals. We then measured T3 and GC concentrations in 834 Steller sea lion fecal samples collected in 2005 and 2006 from 15 sites (haulouts and rookeries) between British Columbia and the Central Aleutian Islands. Overall, GC concentrations did not differ between haulout populations (western 2006 pre-pupping and eastern 2005 post-pupping). Fecal hard-part analyses revealed a lower energy-content diet in the western population, suggesting that diet quality is a relevant hypothesis to explain slightly higher GC concentrations found in the western population, specifically the Aleutian Islands region. However, nutritional stress could not be substantiated through T3 concentrations. The rookeries possessed the highest energy-content diets, but also exhibited a nutritional stress response with a significantly higher GC and lower T3 concentration than either haulout population (possibly related to lactation or decreased foraging opportunities), but T3 comparisons performed at scales of site and region were inconclusive.

No description available.

Reduced availability of high energy-content prey (nutritional stress) is a predominant hypothesis to explain the decline of Steller sea lion (Eumetopias jubatus) populations in western Alaska from the late 1970'€™s to the late 1990'™s. Animals may physiologically respond to consuming insufficient prey by increasing stress levels and decreasing metabolic rates. It may thus be possible to identify nutritional stress by measuring concentrations of glucocorticoids (stress) and thyroid hormones (metabolism) shed in feces. However, techniques to measure thyroid hormone concentrations from Steller sea lion feces have not been developed. We quantified variation of triiodothyronine (T3) and thyroxine (T4) concentrations in Steller sea lion feces following injections of thyrotropin (TSH) into four captive animals. Glucocorticoids (GC) were also assayed to examine any relationship to stimulated thyroid hormone secretion. We found that fecal T3 peaked 48 h post-injection and increased 25-57% in three sea lions (all animals, p=0.03). Pre-injection GC increases indicated stress from isolation for baseline fecal collections, but post-injection increases could not be confirmed as a response to TSH injections or as a product of the study design. The results demonstrated that pre- and post-injection changes in fecal GC and T3 concentrations were consistent with predictions of an increased stress response and metabolic rate within the animals. We then measured T3 and GC concentrations in 834 Steller sea lion fecal samples collected in 2005 and 2006 from 15 sites (haulouts and rookeries) between British Columbia and the Central Aleutian Islands. Overall, GC concentrations did not differ between haulout populations (western 2006 pre-pupping and eastern 2005 post-pupping). Fecal hard-part analyses revealed a lower energy-content diet in the western population, suggesting that diet quality is a relevant hypothesis to explain slightly higher GC concentrations found in the western population, specifically the Aleutian Islands region. However, nutritional stress could not be substantiated through T3 concentrations. The rookeries possessed the highest energy-content diets, but also exhibited a nutritional stress response with a significantly higher GC and lower T3 concentration than either haulout population (possibly related to lactation or decreased foraging opportunities), but T3 comparisons performed at scales of site and region were inconclusive.

Alpha1-proteinase inhibitor (alpha1-PI) has been shown to be a useful marker of gastrointestinal protein loss in some species. The objectives of this study were, first, to develop and analytically validate an ELISA for the measurement of alpha1-PI in feces and serum from cats, and, second, to evaluate fecal alpha1-PI concentrations in healthy cats and cats with chronic gastrointestinal disease. The lower detection limits of the ELISA were 0.02 g/L for serum and 0.04 microgram/gram for feces. The observed-to-expected (O/E) ratios for serial dilutions of serum and fecal samples ranged from 100.0 to 129.7% (mean +/- SD: 112.2 +/- 9.9%) and 103.5 to 141.6% (115.6 +/- 12.8%), respectively. The O/E ratios for samples spiked with seven known concentrations of alpha1-PI ranged from 82.3 to 107.8% (94.7 +/- 7.6%) for serum and 78.5 to 148.7% (96.8 +/- 18.2%) for feces. The coefficients of variation for intra-assay and inter-assay variability were <7.9% and &lt;12.1% for serum, and 5.3%, 11.8%, and 14.2% and 7.7%, 10.2%, and 20.4% for feces, respectively. Reference intervals were 0.6 to 1.4 g/L for serum and up to 1.6 microgram/g for feces. We conclude that this ELISA is sufficiently linear, accurate, precise, and reproducible. For the clinical evaluation, twenty cats with clinical signs of chronic gastrointestinal disease and 20 healthy control cats were enrolled. The diseased cats were grouped into two groups: mild to moderate idiopathic inflammatory bowel disease (IBD) (Group A; n=8) and severe IBD or neoplastic disease (Group B; n=12), based on histopathology results of endoscopic biopsies. Fecal alpha1-PI concentrations and serum concentrations of total protein, albumin, globulin, cobalamin, folate, pancreatic lipase immunoreactivity, and trypsin-like immunoreactivity were determined. Nineteen of the 20 diseased cats had increased fecal alpha1-PI concentrations, ranging from 1.9 to 233.6 microgram/g (normal range: <= 1.6 microgram/g). Fecal alpha1-PI concentrations were statistically significantly different between healthy cats and cats of Group A (median: 3.9 microgram/g, range: 1.3 to 9.2 microgram/g, P<0.001) or cats of Group B (median: 20.6 microgram/g, 4.3 to 233.6 microgram/g; P<0.001), and also between cats of Groups A and B (P<0.01). Hypoalbuminemia, hypoproteinemia, and hypocobalaminemia were detected in 88%, 83%, and 56% of the diseased cats, respectively. Our study suggests that increased fecal alpha1-PI concentrations in association with hypoalbuminemia may be a common finding in cats with IBD or GI neoplasia. Furthermore, alpha1-PI concentrations appear to be higher in cats with severe IBD or confirmed GI neoplasia when compared to cats with mild to moderate IBD.