[Show abstract][Hide abstract] ABSTRACT: Among grow-to-finish pigs from 10 herds in Alberta and Saskatchewan, 23 (16%) of 144 fecal samples were culture-positive and 40 (28%) of 144 pigs were seropositive for Salmonella. With a Bayesian model specifying dependence between the 2 tests, the sensitivity (Se) of culture and real-time polymerase chain reaction (RT-PCR) was 79% to 86%, depending on the cut-off value for the enzyme-linked immunosorbent assay (ELISA). Culture specificity (Sp) was assumed to be 100%; RT-PCR Sp was found to be 94%. The ELISA Se was 76% and 51% at optical density cut-off values ≥ 20% and ≥ 40%, respectively; the Sp was 94% at each cut-off value. The model showed some sensitivity to ELISA prior information, the ELISA Se being approximately 8% lower when informative prior information was specified in the model. When there was no adjustment for dependence between culture and RT-PCR, the posterior estimates for both culture and RT-PCR Se were 11% higher than with the conditional-dependence model and had considerably narrower probability intervals, which suggests that correlation between culture and PCR is important and should be adjusted for in future studies.
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire 10/2011; 75(4):308-11. · 1.02 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The study objectives were to investigate Salmonella prevalence, serovar distribution, and risk factors for shedding in 10 purposively selected farrow-to-finish farms in Saskatchewan and Alberta. Pooled fecal samples from the breeding and grow-finish phases and individual fecal samples from breeding, nursery, and grow-finish pigs were cultured for Salmonella; serotyping of isolates was performed. Pig and pen characteristics were recorded for each pig and pen sampled.Overall, 407/1143 (36%) of samples were Salmonella positive; within-farm prevalence ranged from 1% to 79%. Sows, nursery, and grow-finish pigs accounted for 43%, 29%, and 28% of positive samples, respectively. More Salmonella were detected in pooled pen than individual pig samples (P < 0.001). Among 418 Salmonella isolates, there were 19 distinct serovars; the most common were S. Derby (28.5%), S. Typhimurium, var. Copenhagen (19.1%), S. Putten (11.8%), S. Infantis (6.8%), and S. Mbandaka (6.1%). Sows were more likely to shed Salmonella than nursery or grow-finisher (OR 2.9, P < 0.001) pigs. Pelleted feed (OR 8.2, P < 0.001) and nose-to-nose pig contact through pens (OR 2.2, P = 0.005) were associated with increased Salmonella prevalence. Significant differences in serovar distribution were detected among production phases. The use of pooled pen samples is recommended as a more efficient means for accurate evaluation of Salmonella status in different phases of pig production. The breeding herd might be an important source of Salmonella persistence within farrow-to-finish farms and should be targeted in control efforts. The latter might also apply to the use of pelleted feed, which remains the most consistently reported significant risk factor for Salmonella shedding in pigs.
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire 04/2010; 74(2):81-90. · 1.02 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The objective of this study was to estimate the apparent and true prevalence of exposure to Toxoplasma gondii in Ontario finisher pigs. During the study period (2001 to 2004), sera from 6048 pigs were tested with a commercial enzyme-linked immunosorbent assay (ELISA); 103 farms were included 1 to 3 times in the study. True prevalence was estimated using a Bayesian approach. Apparent prevalence at the pig level was 1.59% [95% confidence interval (CI): 0.45, 2.99] in 2001, 0.06% (95% CI: 0.00, 0.46) in 2003, and 0.26% (95% CI: 0.00, 0.82) in 2004. Apparent prevalence at the herd-level was 13.7% (95% CI: 7.5, 22.3) in 2001; 1.25% (95% CI: 0.03, 6.77) in 2003, and 3.75% (95% CI: 0.78, 10.6) in 2004. Similarly, posterior Bayesian estimates of true prevalence at the pig level were 1.7% [95% probability interval (PI): 1.2, 2.2] in 2001, 0.2% (95% PI: 0.04, 0.4) in 2003, and 0.3% (95% PI: 0.1, 0.7) in 2004. At the herd level, posterior estimates of prevalence were 11.6% (95% PI: 7.4, 16.8) in 2001, 0% (95% PI: 0.0, 2.5) in 2003, and 1.2% (95% PI: 0.0, 5.0) in 2004 when a herd cut-point > or = 1 was used. Exposure to T. gondii in finishing pig farms in Ontario appears to be infrequent.
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire 08/2008; 72(4):303-10. · 1.02 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A province-wide cross-sectional seroprevalence and agroecological risk factor study of Mycobacterium avium subspecies paratuberculosis (MAP) and Neospora caninum (NC) infection among cattle in 100 cow-calf herds in Alberta was conducted. The seroprevalence of MAP in adult cattle was 1.5% across all herds. Using a widely accepted herd test cutpoint of 2 or more seropositive cows out of 30 animals tested, 7.9% of herds were estimated to be infected (95% confidence interval (CI): 2.3-23.4%). Seroprevalence of MAP differed by agroecological region; specifically, cattle and herds in areas with high soil pH (> 7.0), southern latitudes, and arid climates had a moderately reduced risk of infection (P < 0.10). Seroprevalence of NC infection was 9.7% among adult beef cattle province-wide--these levels also varied by agroecological region--with 91.0% of herds infected overall.
The Canadian veterinary journal. La revue veterinaire canadienne 04/2007; 48(4):397-406. · 0.52 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The aim of this study was to determine serological prevalence for Salmonella in 90 Alberta finishing swine farms over a 5-month period; to evaluate the correlation between the detection of Salmonella by bacteriological culture and serology; and to identify risk factors for Salmonella seroprevalence. Participating farms were visited 3 times. A total of 30 blood and 15 fecal samples were collected from finishing pigs on each farm. VetScreen Salmonella covalent mix-ELISA (Svanovir) and conventional culture were performed. The apparent Salmonella seroprevalences at the sample and farm level were 13.2% (95% confidence interval [CI], 10.5-15.5%) and 83.3% (95% CI, 74-90.4%), respectively. Most of the farms had within-farm seroprevalence of <or=20%, indicating that pigs on these farms had a low exposure to Salmonella. The Salmonella farm status changed frequently across 3 visits. The correlation between fecal prevalence and seroprevalence at the farm and farm visit level was 0.71 (P < 0.0001) and 0.47 (P < 0.0001), respectively. The use of meal feed and the reported use of antimicrobials through water were associated with a lower farm seroprevalence for Salmonella. Longitudinal sampling and testing are required to properly evaluate Salmonella on-farm status. The interpretation of existing serological and culture tests for Salmonella in swine should take into consideration their imperfect sensitivity, what these tests actually measure (previous exposure vs. current shedding), and the Salmonella serovar distribution within the targeted population. Further work is necessary to demonstrate the effectiveness of on-farm interventions against Salmonella in swine.
[Show abstract][Hide abstract] ABSTRACT: A province-wide, cross-sectional seroprevalence and agroecological risk factor study of Mycobacterium avium subspecies paratuberculosis (MAP), Neospora caninum (NC), Bovine leukemia virus (BLV), and Bovine viral diarrhea virus (BVDv) genotypes 1 and 2 (BVDv1 and BVDv2) infection in dairy cattle herds in Alberta was conducted. Among adults, the seroprevalence of MAP, NC, and BLV was 9.1%, 18.5%, and 26.9%, respectively. For MAP, based on a herd test cutpoint of 2 or more seropositive cows, 58.8% of herds were infected. Herd-level seroprevalence for NC and BLV was 98.7% and 86.7%, respectively, based on a herd-test cutpoint of 1 seropositive cow. Among unvaccinated dairy heifers, the seroprevalence for BVDv1 and BVDv2 infection was 28.4% and 8.9%, respectively, while herd-level infection was 53.4% and 19.7%. Seroprevalence for MAP varied moderately by agroecological region, whereas that for NC, BLV, and BVDv1 and BVDv2 did not. For MAP, aridity and soil pH (correlated features of the region) were also important.
The Canadian veterinary journal. La revue veterinaire canadienne 11/2006; 47(10):981-91. · 0.52 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Enzyme-linked immunosorbent assay (ELISA) and culture are 2 common diagnostic tests for detecting Mycobacterium avium subsp. paratuberculosis (Map) in Johne's disease, but they are not as sensitive as polymerase chain reaction (PCR). However, inhibitors can coextract with the target DNA and cause interference in PCR. Development of an immune capture assay followed by PCR amplification can alleviate this problem. In this study, we were able to induce an immune response in chickens using heat or formalin inactivated Map. The purified immunoglobulin (Ig)Y has a molecular weight of 160 kDa. The titers were at 1:6400 and 1:12 800 at weeks 5 to 6 and 8 to 9, respectively, as determined by the IDEXX modified ELISA kit for Johne's disease. The IgY produced from inactivated bacterial cells had no effect on its ability to recognize live Map cells as illustrated by immunofluorescence assay and immune capture PCR results.
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire 11/2004; 68(4):302-8. · 1.02 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: In this study, a commercial Salmonella covalent mix-enzyme linked immunosorbent assay (ELISA) for serological detection of Salmonella infection in swine was evaluated by comparing it with the conventional fecal culture method and inter-laboratory proficiency testing, using a panel of sera tested in 5 laboratories from Europe and North America. Comparison with culture results showed that 88.5% of 26 culture-positive animals were ELISA positive, as were 55% of 60 animals from 2 culture-positive pig herds. Of 90 animals from 2 high health farms with no clinical symptoms of salmonellosis, 98.9% tested negative. The interlaboratory comparison study found a kappa value of 0.9 between our laboratory (using an automated system) and the manufacturer laboratory (using the manual method). Comparison of ELISA results from all 5 participating laboratories showed very good to excellent agreement, between 85% and 97.5%. We found this assay to be useful for the screening of antibodies against Salmonella present in swine serum. It agrees well with bacterial cultures, is reproducible, sensitive, specific, repeatable, and suitable for automation.
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire 05/2004; 68(2):134-9. · 1.02 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The authors automated an enzyme-linked immunosorbent assay to detect porcine serum antibodies to Toxoplasma gondii. Two thousand swine sera can be assayed in two eight-hour shifts using a robotic workstation. The automated-ELISA programming is not complex and the test configuration is flexible. This high-throughput screening (HTS) a-ELISA can achieve a 10-fold increase (100→1000 tests) in test capacity over the manual method. The assay has been validated according to the requirements of the ISO/IEC 17025 standard. These include repeatability, reproducibility, and optimal threshold value studies. Other requirements are proficiency panel testing, analyst training, standard operation procedure, and equipment certifications.
Journal of the Association for Laboratory Automation 11/2003; 8(5):42-50. DOI:10.1016/S1535-5535(03)00008-X · 1.50 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This is a validation study of 2 commercially available enzyme linked immunosorbent assays (ELISA) for the detection of antibodies against Neospora caninum in bovine serum. The results of the reference sera (n = 30) and field sera from an infected beef herd (n = 150) were tested by both ELISAs and the results were compared statistically. When the immunoblotting results of the reference bovine sera were compared to the ELISA results, the same identity score (96.67%) and kappa values (K) (0.93) were obtained for both ELISAs. The sensitivity and specificity values for the IDEXX test were 100% and 93.33% respectively. For the Biovet test 93.33% and 100% were obtained. The corresponding positive (PV+) and negative predictive (PV-) values for the 2 assays were 93.75% and 100% (IDEXX), and 100% and 93.75% (Biovet). In the 2nd study, competitive inhibition ELISA (c-ELISA) results on bovine sera from an infected herd were compared to the 2 sets of ELISA results. The identity scores of the 2 ELISAs were 98% (IDEXX) and 97.33% (Biovet). The K values calculated were 0.96 (IDEXX) and 0.95 (Biovet). For the IDEXX test the sensitivity and specificity were 97.56% and 98.53%, whereas for the Biovet assay 95.12% and 100% were recorded, respectively. The corresponding PV+ and PV- values were 98.77% and 97.1% (IDEXX), and 100% and 94.44% (Biovet). Our validation results showed that the 2 ELISAs worked equally well and there was no statistically significant difference between the performance of the 2 tests. Both tests showed high reproducibility, repeatability and substantial agreement with results from 2 other laboratories. A quality assurance based on the requirement of the ISO/IEC 17025 standards has been adopted throughout this project for test validation procedures.
Canadian journal of veterinary research = Revue canadienne de recherche vétérinaire 11/2002; 66(4):264-71. · 1.02 Impact Factor