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41.
The aims of this study were to develop a new real-time quantitative PCR (QPCR) assay based on IS900 for detection and quantification of Mycobacterium avium subsp. paratuberculosis (MAP) DNA in faeces, and to use this to detect infected sheep. Both the C and S strains of MAP were detected by the QPCR assay, and no cross reactions were detected with 51 other species of mycobacteria including 10 which contained IS900-like sequences. One copy of IS900 fragment cloned into plasmid pCR2.1 and 1 fg of MAP genomic DNA were consistently detected, while in spiked faecal samples the detection limit was 10 viable MAP per gram of ovine faeces. A total of 506 individual ovine faecal samples and 27 pooled ovine faecal samples with known culture results were tested. The QPCR assay detected 68 of 69 BACTEC culture positive individual faeces and there was a strong relation between time to detection in culture and DNA quantity measured by QPCR (r= -0.70). In pooled faecal samples, QPCR also agreed with culture (kappa=0.59). MAP DNA was detected from some culture negative faecal samples from sheep exposed to MAP, suggesting that the QPCR has very high analytical sensitivity for MAP in faecal samples and detects non-viable MAP in ovine faeces. None of the faecal samples from 176 sheep that were not exposed to MAP were positive in QPCR. This is the first report of a direct faecal QPCR assay that has similar sensitivity to a gold standard radiometric culture assay. 相似文献
42.
Eamens GJ Whittington RJ Turner MJ Austin SL Fell SA Marsh IB 《Veterinary microbiology》2007,125(1-2):22-35
Dilution rates for pooled faecal culture (PFC) and direct IS900 polymerase chain reaction (D-PCR) tests were evaluated on faecal samples from infected cows mixed with uninfected faeces in dilutions from 1 in 5 to 1 in 50. PFC was performed by radiometric culture, with confirmation by IS900 PCR and restriction endonuclease analysis (PCR/REA) on growth, and by mycobactin dependency testing on solid medium. Using 37 culture positive faecal samples from 12 subclinical cows, 83.8% and 94.6% of samples were confirmed positive in the PFC assay at dilutions of 1 in 50 and 1 in 30, respectively. Lower dilutions (1 in 5 to 1 in 20) provided only marginally better sensitivity, and confirmation of PFC growth by PCR/REA was significantly more sensitive than mycobactin dependency. D-PCR had significantly lower sensitivity than PFC confirmed by PCR/REA, with pools of 1 in 50, 30, 10 and 5 yielding positive results in 64.9%, 70.3%, 78.4% and 83.8% of samples, respectively. Cattle considered to be shedding 1.5 x 10(6) viable M. avium subsp. paratuberculosis (Map)/g faeces (on the basis of estimated losses in processing and growth rates in radiometric broth) were positive at dilutions up to 1 in 50 in the PFC and D-PCR. Those shedding 5 x 10(5) viable Map/g were positive in the PFC at dilutions up to 1 in 40, but required a 1 in 10 dilution or less for D-PCR. The results suggest that for cattle shedding relatively high concentrations of Map in faeces (>2 x 10(5) viable Map/g), maximal dilutions of 1 in 30 for PFC and 1 in 10 for D-PCR would be applicable. 相似文献
43.
The repeatability of detection of Mycobacterium avium subsp. paratuberculosis (Map) within and between samplings from 16 paratuberculous dairy cows (13 subclinical; 3 clinical) was investigated by radiometric culture of quadrants of faecal dung pats collected on four to seven occasions over a 10-16-day period. Results were compared to serological status and to pathological and bacteriological findings in multiple tissues obtained at slaughter from 15 of the animals 2-6 weeks after the faecal samplings. From faecal samples taken on 77 occasions over the 2-week period, 296/308 (96%) quadrants were culture positive, with samples from all cattle showing evidence of faecal shedding of Map. Histological lesions typical of paratuberculosis were present in 14 of the 15 cows examined at slaughter, varying in severity from mild (two animals) to moderate (4) and advanced (8), and all predilection tissue sites yielded Map. The negative faecal samples were derived from a single animal that was culture positive in two quadrants on each of the first two (of four) sampling occasions (i.e. culture positive in only 4 of 16 collected quadrants). This animal was found to be histologically negative at slaughter, and culture positive from three of five predilection tissue sites. Faecal samples from cows with subclinical and clinical paratuberculosis, with lesion severity ranging from mild to severe at multiple predilection sites, produced faeces with relatively consistent concentrations of Map within samples. There was significant variation in concentrations of Map between samples in individual animals over a period of 2 weeks, but this did not affect the dichotomous positive-negative culture status for 15 of the 16 cattle. A faecal sample collected non-randomly per rectum thus provides a representative specimen for detection of Map by radiometric culture on a single sampling occasion. 相似文献
44.
Goldstein RE Atwater DZ Cazolli DM Goldstein O Wade CM Lindblad-Toh K 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2007,21(1):199-203
BACKGROUND: Primary hyperparathyroidism (PHPT) is caused by inappropriate secretion of parathyroid hormone (PTH) by autonomously functioning neoplastic or hyperplastic parathyroid "chief" cells. Keeshonden are thought to be over-represented in studies on canine PHPT, but no proof of heritability or mode of inheritance has been published. The canine disease clinically resembles human familial isolated hyperparathyroidism (FIHP). HYPOTHESIS: Primary hyperparathyroidism in Keeshonden is genetically transmitted and is caused by a mutation in 1 of 4 genes implicated in human FIHP: MEN1, CASR, HRPT2, or RET. ANIMALS: Pedigrees consisting of 1647 Keeshonden were created including 219 Keeshonden with known PHPT phenotypes (69 positive). DNA samples were obtained from 176 of the 219 Keeshonden (34 positive). METHODS: Heritability and mode of inheritance were determined by segregation analysis. Canine homologs to the human genes were identified. Exons and surrounding intron regions were sequenced and scanned for sense-altering polymorphisms or polymorphisms that segregated with the disease. Messenger RNA from a parathyroid tumor of an affected Keeshond was analyzed for polymorphisms and splice alterations. RESULTS: PHPT follows an autosomal dominant mode of inheritance in Keeshonden with possible age-dependent penetrance. No polymorphisms identified in the genes analyzed were associated with a change in predicted protein or in hypothesized splice sites. CONCLUSIONS AND CLINICAL IMPORTANCE: PHPT is an autosomal dominant, genetically transmitted disease in Keeshonden. Once the mutation locus is identified, genetic testing should quickly decrease the incidence of PHPT in this breed. It is unlikely that mutations in MEN1, CASR, HRPT2, or RET cause PHPT in Keeshonden. 相似文献
45.
An orbital sialocele developed in a dog following enucleation for protracted glaucoma. This eye had historically been treated for keratoconjunctivitis sicca by parotid duct transposition approximately 5 years previously, and the duct was ligated distally at the time of enucleation. One month following enucleation, the dog presented with a fluctuant conical-shaped swelling ventrolateral to the orbital socket. Surgical exploration revealed a dilated, fibrotic distal portion of the previously transposed parotid duct, and saliva, within the enucleated orbit. The distal portion of the duct and saliva-containing tissues from within the orbit were excised. The remaining proximal normal portion of the parotid duct was re-routed into the oral cavity. Clinicopathologic and histologic examination of the excised orbital contents and dilated portion of duct revealed a sterile sample of saliva and moderate chronic periductal fibrosis. At a 6-month re-evaluation there was no evidence of recurrence of the sialocele, and the parotid duct was functional. 相似文献
46.
Ticehurst K Zaki S Maddern K Lingard A Barrs V Malik R 《Journal of Feline Medicine and Surgery》2007,9(6):521-525
A 14-year-old neutered male domestic shorthaired cat was presented to the University Veterinary Centre Sydney for evaluation and treatment of dental disease. This cat developed an unusual bradyarrhythmia under anaesthesia. The possible causes and treatment of the dysrythmia are discussed. 相似文献
47.
Burkitt JM Haskins SC Nelson RW Kass PH 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2007,21(2):226-231
BACKGROUND: A syndrome of relative adrenal insufficiency has been identified in septic humans, and is associated with hypotension and death. Relative adrenal insufficiency is generally associated with basal serum cortisol concentration within or above the reference range and a blunted cortisol response to adrenocorticotropic hormone administration. It is unknown whether relative adrenal insufficiency occurs in septic dogs. HYPOTHESIS: That relative adrenal insufficiency occurs in septic dogs, and that relative adrenal insufficiency is associated with hypotension and mortality. ANIMALS: Thirty-three septic dogs admitted to a small animal intensive care unit. METHODS: Dogs were included in the study if they had a known or suspected infectious disease and had systemic inflammatory response syndrome. Dogs were excluded if they had disease or medication history expected to affect the hypothalamic-pituitary-adrenal axis. Serum cortisol and endogenous plasma adrenocorticotropic hormone concentrations were measured before, and serum cortisol concentration measured 1 hour after, intramuscular administration of 250 microg of cosyntropin/dog. The change in cortisol concentration (delta-cortisol) before and after cosyntropin administration was determined in each dog. RESULTS: Hypotension was associated with lower delta-cortisol values (OR 1.3; CI 1.0-1.9; P = .029). delta-Cortisol cutoff of 3.0 microg/dL was most accurate for predicting hypotension, survival to discharge, and 28-day survival. The rate of death in dogs with delta-cortisol < or = 3 microg/dL was 4.1 times that of dogs with delta-cortisol > 3 microg/dL (RR 4.1; CI 1.5-12.3; P = .01). CONCLUSIONS AND CLINICAL RELEVANCE: Delta-cortisol < or = 3 microg/dL after adrenocorticotropic hormone administration is associated with systemic hypotension and decreased survival in septic dogs. 相似文献
48.
A dynamic optimization model was developed and used to evaluate alternative foot-and-mouth disease (FMD) control strategies. The model chose daily control strategies of depopulation and vaccination that minimized total regional cost for the entire epidemic duration, given disease dynamics and resource constraints. The disease dynamics and the impacts of control strategies on these dynamics were characterized in a set of difference equations; effects of movement restrictions on the disease dynamics were also considered. The model was applied to a three-county region in the Central Valley of California; the epidemic relationships were parameterized and validated using the information obtained from an FMD simulation model developed for the same region. The optimization model enables more efficient searches for desirable control strategies by considering all strategies simultaneously, providing the simulation model with optimization results to direct it in generating detailed predictions of potential FMD outbreaks. 相似文献
49.
Malik R 《The Journal of small animal practice》2007,48(8):474; author reply 474
50.