Comparison of milk culture, direct and nested polymerase chain reaction (PCR) with fecal culture based on samples from dairy herds infected with Mycobacterium avium subsp. paratuberculosis          下载免费PDF全文

Anli Gao  Joseph Odumeru  Melinda Raymond  Steven Hendrick  Todd Duffield    Lucy Mutharia 《Canadian journal of veterinary research》2009,73(1):58-64

Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne’s disease in cattle and other farm ruminants, and is also a suspected pathogen of Crohn’s disease in humans. Development of diagnostic methods for MAP infection has been a challenge over the last few decades. The objective of this study was to investigate the relationship between different methods for detection of MAP in milk and fecal samples. A total of 134 milk samples and 110 feces samples were collected from 146 individual cows in 14 MAP-infected herds in southwestern Ontario. Culture, IS900 polymerase chain reaction (PCR) and nested PCR methods were used for detecting MAP in milk; results were compared with those of fecal culture. A significant relationship was found between milk culture, direct PCR, and nested PCR (P < 0.05). The fecal culture results were not related to any of the 3 assay methods used for the milk samples (P > 0.10). Although fecal culture showed a higher sensitivity than the milk culture method, the difference was not significant (P = 0.2473). The number of MAP colony-forming units (CFU) isolated by culture from fecal samples was, on average, higher than that isolated from milk samples (P = 0.0083). There was no significant correlation between the number of CFU cultured from milk and from feces (Pearson correlation coefficient = 0.1957, N = 63, P = 0.1243). The animals with high numbers of CFU in milk culture may not be detected by fecal culture at all, and vise versa. A significant proportion (29% to 41%) of the positive animals would be missed if only 1 culture method, instead of both milk and feces, were to be used for diagnosis. This suggests that the shedding of MAP in feces and milk is not synchronized. Most of the infected cows were low-level shedders. The proportion of low-level shedders may even be underestimated because MAP is killed during decontamination, thus reducing the chance of detection. Therefore, to identify suspected Johne’s-infected animals using the tests in this study, both milk and feces samples should be collected in duplicate to enhance the diagnostic rate. The high MAP kill rate identified in the culture methods during decontamination may be compensated for by using the nested PCR method, which had a higher sensitivity than the IS900 PCR method used.  相似文献   

In vitro Maturation of Oocytes from Santa Ines Ewes Subjected to Consecutive Sessions of Follicular Aspiration by Laparoscopy     

LC Padilha  PPM Teixeira  EA Pires‐Buttler  M Apparício  TF Motheo  PAP Savi  EYO Nakaghi  AE Alves  WRR Vicente 《Reproduction in domestic animals》2014,49(2):243-248

The success of embryo production in vitro depends upon the use of an efficient oocyte retrieval technique, and the best results have been obtained by laparoscopic aspiration. The aim of this study was to evaluate the effect of consecutive sessions of follicular aspiration on the quantity, quality and in vitro maturation competence of oocytes obtained from ewes subjected to hormonal stimulation. Six Santa Ines ewes underwent nine sessions of follicular aspiration by laparoscopy with a 7‐day interval between sessions, totalling 56 aspirations. After 24 h of culture, oocytes were stained and classified according to the stage of nuclear and cytoplasmic maturation. Oocyte retrieval rate was 61.4 ± 2%, resulting in a total of 249 oocytes. No significant variation was observed between sessions (p > 0.05). The average number of oocytes retrieved from each ewe was 6.4 ± 2 per session and 42 ± 4 in total. No significant difference was observed between the frequencies of the different stages of nuclear maturation: 32.72% mature, 40.74% immature and 26.54% degenerated/indeterminate oocytes; however, a significant difference was observed between the frequencies of the different stages of cytoplasmic maturation: 10.7% mature, 73.25% immature and 16.05% degenerated/indeterminate oocytes. No significant difference was observed in nuclear or cytoplasmic maturation between the weeks of procedure. We conclude that after nine consecutive sessions of follicular aspiration, the quantity and quality of retrieved oocytes remained unchanged as well as the levels of nuclear and cytoplasmic maturation obtained, demonstrating the viability of this technique for repetitive follicular aspirations on the same donor.  相似文献   

Ketosis Treatment in Lactating Dairy Cattle     

Jessica L. Gordon  Stephen J. LeBlancTodd F. Duffield  DVM  DVSc 《Veterinary Clinics of North America: Food Animal Practice》2013

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Effect of paratuberculosis on culling, milk production, and milk quality in dairy herds     

Hendrick SH  Kelton DF  Leslie KE  Lissemore KD  Archambault M  Duffield TF 《Journal of the American Veterinary Medical Association》2005,227(8):1302-1308

OBJECTIVE: To determine the effect of paratuberculosis on culling, milk production, and milk quality in infected dairy herds. DESIGN: Cross-sectional study. ANIMALS: 689 lactating dairy cows in 9 herds. PROCEDURE: Milk, blood, and fecal samples were obtained from all cows. Fecal samples were evaluated via mycobacterial culture. Serum samples were tested with a commercially available ELISA for antibodies against Mycobacterium avium subsp paratuberculosis, and preserved milk samples were tested with an ELISA for antibodies against M paratuberculosis. Mixed effect and proportional hazards models were used to determine the effect of paratuberculosis on 305-day milk, fat, and protein production; somatic cell count linear score; and the risk of culling. RESULTS: Cows with positive results of bacteriologic culture of feces and milk ELISA produced less milk, fat, and protein, compared with herdmates with negative results. No difference in 305-day milk or fat production was detected in cows with positive results of serum ELISA, compared with seronegative cows. The 3 survival analyses revealed that cows with positive results of each test were at higher risk of being culled than cows with negative results. Paratuberculosis status, as determined by use of all 3 diagnostic tests, was not associated with milk somatic cell count linear score. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that for the 9 herds in this study, paratuberculosis significantly decreased milk production and cow longevity.  相似文献   

Multiattribute evaluation of two simple tests for the detection of Cryptosporidium parvum in calf faeces     

Trotz-Williams LA  Martin SW  Martin D  Duffield T  Leslie KE  Nydam DV  Jamieson F  Peregrine AS 《Veterinary parasitology》2005,134(1-2):15-23

There is a need for simple and inexpensive diagnostic and screening tests for the detection of Cryptosporidium parvum infection in calves. A sucrose wet mount test and a lateral immunochromatography test were evaluated for epidemiological sensitivity and specificity, cost per test, simplicity, test time and ease of batching. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the Cryptosporidium oocyst wall protein (COWP) gene locus, with gel electrophoresis, was used as a gold standard. Cohen's kappa statistic of agreement (kappa) between the Ontario Veterinary College (OVC) sucrose wet mount test and COWP PCR-RFLP was 0.82, and the sensitivity and specificity of the OVC sucrose wet mount test were 88.6% and 93.8%, respectively. The sensitivity and specificity of the lateral immunochromatography test were 78.3% and 93.3%, respectively, and agreement between this test and PCR-RFLP was good (kappa=0.73). There was substantial agreement between the OVC sucrose wet mount test and the lateral immunochromatography test (kappa=0.84). Both tests were inexpensive and easy to use; however, the lateral immunochromatography test was faster and simpler to perform than the sucrose wet mount test, and was generally more user-friendly. These tests provide practitioners and researchers with cheap, quick and accurate methods of detecting C. parvum infection in young calves.  相似文献   

Diagnosis of botulism in cattle using ELISA to detect antibody to botulinum toxins   总被引：1，自引：0，他引：1  

TF JUBB  TM ELLIS  AR GREGORY 《Australian veterinary journal》1993,70(6):226-227

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The development and evaluation of an enzyme-linked immunosorbent assay for the detection of Mycobacterium bovis     

B J Duffield 《Veterinary microbiology》1990,24(2):205-209

A double antibody sandwich layer enzyme-linked immunosorbent assay (ELISA) was used to detect Mycobacterium bovis. The ELISA detected M. bovis is pure culture at concentrations of 1 x 10(5) colony-forming units (CFU) ml-1 and greater, compared to a minimum detection level of 1 x 10(6) CFU ml-1 for isolation techniques. Neither technique detected M. bovis at 1 x 10(4) CFU ml-1. The ELISA did not cross-react with common mycobacterial contaminants such as Mycobacterium avium intracellulare-scrofulaceum complex serotypes 18 and 42, M. terrae, M. fortuitum, M. flavescens and with Escherichia coli or Rhodococcus equi. Further work is needed to evaluate this assay in detecting M. bovis in tissues and the environment.  相似文献   

Intestinal atresia in Friesian calves     

TF JUBB 《Australian veterinary journal》1990,67(10):382-382

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Prognostic factors for recovery from coxo-femoral dislocation in cattle     

TF. JUBB  J. MALMO†  P. BRIGHTLING  GA. ANDERSON  GM. DAVIS† 《Australian veterinary journal》1989,66(11):354-358

Details were recorded of 47 cases of coxo-femoral dislocation observed in cattle over 1 year. Treatment was successful in 20/47 (42.6%) cases using a method of closed reduction. Factors that most strongly influenced the prognosis were identified. The most useful single prognostic factor was whether the cow was able to stand before reduction. Other factors that also had a strong positive influence on the prognosis were: age less than 3y, bodyweight less than 400kg and duration of dislocation less than 12h. Nineteen unsuccessful cases were examined at the knackery. Four were found to have a fracture of the proximal femur. The coxo-femoral joint was carefully dissected in the other 15 cases and there was no evidence of hip dysplasia. A seasonal incidence of dislocations, which coincides with the calving and mating periods, was demonstrated during the survey and from practice records for the previous 3 years.  相似文献   

The impact of controlled release capsules of monensin on postcalving haptoglobin concentrations in dairy cattle          下载免费PDF全文

Rex G. Crawford  Ken E. Leslie  Randal Bagg  C. Paul Dick    Todd F. Duffield 《Canadian journal of veterinary research》2005,69(3):208-214

This study was designed to assess the impact of a controlled release capsule (CRC) of monensin, administered prior to calving, on postcalving haptoglobin levels. The role of disease on haptoglobin levels was also studied. The study population consisted of 1010 cows from 25 Holstein dairy herds near Guelph, Ontario. Monensin CRC or placebo capsules were randomly assigned within each herd 3 wk prior to the expected calving date. Serum from week 1 and week 6 postcalving was submitted for quantification of haptoglobin concentrations. Haptoglobin results were analyzed for associations with treatment, health data, and individual cow factors up to 95 d in milk. Haptoglobin concentrations were higher in week 1 than week 6 (P < 0.05). In univariate analysis, several diseases were significantly associated with haptoglobin concentrations. However, occurrence of disease appeared to be a confounding factor in the data interpretation. Thus, the analysis was stratified by the presence or absence of disease. There appeared to be associations between factors other than clinical disease contributing to increased haptoglobin levels in both clinically healthy and unhealthy cattle. Haptoglobin served as a good indicator of inflammatory disease. Monensin CRC treatment was associated with increased haptoglobin concentrations in clinically unhealthy cattle, perhaps reflecting a better ability to respond to disease challenge. The lower haptoglobin concentrations in monensin CRC treated cattle that were clinically normal may be a reflection of reduced subclinical disease.  相似文献