Randomized control trial to investigate compliance with, and impacts of, cow comfort recommendations on smallholder dairy farms in Kenya.

Smallholder dairy farmers (SDF) in developing countries often have limited understanding on the importance of cow comfort. We conducted a randomized controlled trial with 124 cows on 114 Kenyan SDF to determine the status of cow comfort, to assess compliance to farm-specific cow comfort recommendations, and to evaluate the impacts of the farm-specific interventions on cow comfort. On the first farm visit, stall dimensions and characteristics (e.g. stall base hardness and hygiene) were measured and categorized as adequate, marginal or inadequate/absent based on cow size. Where measurements were not adequate, farm-specific cow comfort recommendations were provided in written and oral form to the randomly allocated intervention group of farms (n = 74). On the second farm visit two months later, the same measurements were taken, and percent compliance to the recommendations was evaluated. A discomfort index was arithmetically calculated based on the stall base hardness (scale was 1-3 for soft to hard) and hygiene (scale was 1-5 for clean to dirty). Multivariable linear regression models were used to determine specific associations with the discomfort index. On the first visit, the mean stall base hardness and stall hygiene scores were 1.7 and 2.3, respectively, for a mean discomfort index of 4.0. Intervention farmers were given 3.9 comfort recommendations, on average, and complied with 2.1 recommendations, significantly improving the discomfort index at visit two to 3.3. The overall compliance to the recommendations was 49.0%. In a final model, the interaction between intervention group and visit number was significantly associated with discomfort index, indicating that after adjusting for baseline discomfort indices, the intervention led to better cow comfort. Specifically, bedding type and neck rail positioning were significantly associated with discomfort index. We concluded that farmers can substantially improve cow comfort on SDF by providing recommendations to them. Farm advisors should include cow comfort recommendations to SDF.

Effects of Calliandra and Sesbania on Daily Milk Production in Dairy Cows on Commercial Smallholder Farms in Kenya.

There is a growing interest in protein supplementation of dairy-cow diets using leguminous shrubs. The study objective was to ascertain the association between diet supplementation with Calliandra calothyrsus and Sesbania sesban and milk production in dairy cattle on commercial smallholder farms. This trial involved 235 cows from 80 smallholder dairy farms in Kenya randomly allocated to 4 intervention groups: (1) receiving Calliandra and Sesbania and nutritional advice; (2) receiving reproductive medicines and advice; (3) receiving both group 1 and 2 interventions; and (4) receiving neither intervention. Farm nutritional practices and management data were collected in a questionnaire, and subsequent physical examinations, mastitis tests, and milk production of cows on the farm were monitored approximately monthly for 16 months. Descriptive and univariable statistical analyses were conducted, and multivariable mixed-model regression was used for identification of factors associated (P < 0.05) with daily milk production. The mean milk production was 6.39 liters/cow/day (SD = 3.5). Feeding Calliandra/Sesbania to cows was associated (P < 0.0005) with an increase in milk produced by at least 1 liter/cow/day with each kg fed. Other variables positively associated with ln daily milk production in the final model included feeding of Napier grass, amount of silage and dairy meal fed, body condition score, and appetite of the cow. Other variables negatively associated with ln daily milk production in the final model included amount of maize germ fed, days in milk, sudden feed changes, pregnancy, and subclinical mastitis. In conclusion, our field trial data suggest that use of Calliandra/Sesbania through agroforestry can improve milk production in commercial smallholder dairy farms in Kenya. Agroforestry land use systems can be adopted as a way for dairy farmers to cope with feed shortages and low crude protein in farm-available feeds for their cows.

Effects of Calliandra and Sesbania supplementation on weight gain in dairy calves on smallholder farms in Kenya.

The growth rate of female calves on dairy farms is one of the crucial factors which influence age at first calving, affecting lifetime lactation productivity of a dairy cow. Diets with adequate crude protein are necessary to support calf growth. The study objective was to determine the effects of nutritional advice and diet supplementation with Calliandra calothyrsus and Sesbania sesban on average daily weight gain in dairy calves on semi-commercial smallholder dairy farms in Kenya. This trial involved 155 calves from 73 smallholder dairy farms, in Kenya, randomly selected and allocated to either intervention or control groups. The intervention group received nutritional advice and seedlings of Calliandra calothyrsus and Sesbania sesban, while the control group did not receive these interventions. Every 1-2 months for 16 months, data on farm nutritional practices and management were collected in a questionnaire, and physical examinations were done to monitor weight and health status. Descriptive and univariable statistical analyses were conducted, and multivariable mixed linear regression models were used for identification of factors associated (P < 0.05) with the natural log transformation of weight gain of calves on a given farm, controlling for clustering of visits within calves. Feeding at least 0.2 kg/day (wet weight) of Calliandra or Sesbania to calves <6 months old resulted in 33.2% increase in daily weight gain, while controlling for confounding by breed and sex of the calf. For calves ≥6 months, there was a significant interaction between amount of hay fed and if calves were also fed on Calliandra or Sesbania. When no Calliandra or Sesbania supplementation was provided, the mean weight gain was low and relatively constant even with increasing amounts of hay. When Calliandra or Sesbania supplement was added to the diet, the mean weight gain increased from 0.17 kg to 0.48 kg when hay was fed at 1 and 5 kg, respectively, while controlling for confounding by amount of maize silage fed and the prevailing season. In conclusion, supplementation of calf diets with at least 0.2 kg/calf/day of Calliandra or Sesbania is beneficial for faster weight gains.

Risk factors associated with cows' lying time, stall and cows' own cleanliness in smallholder dairy farms in Kenya.

BACKGROUND AND AIM: The welfare of animals kept in livestock production systems has raised concerns around the world. Adult dairy cattle require adequate rest and spend approximately 12 h/day lying down. This cross-sectional study aimed to determine the stall factors and management practices affecting cows' lying time, stall cleanliness, and cows' cleanliness (udder and upper leg), in smallholder dairy cows in Meru County of Kenya. MATERIALS AND METHODS: A total of 106 milking cows from 73 farms were assessed for daily lying time and cleanliness. Data loggers were used to record the lying time of cows for 3 days. Stall, udder, and upper leg cleanliness were assessed using a 5-score system: 1 (very clean) to 5 (very dirty). Management information was acquired using a questionnaire that was administered face-to-face to the farmers in their native Kimeru language. Univariable and multivariable linear and logistic regression models were fit to determine factors associated with cows' lying time and dichotomized stall and cows' own cleanliness, respectively. RESULTS: The mean daily lying time was 10.9±2.2 h, and the mean stall cleanliness score was 2.4±1.0. The mean average cleanliness scores of the udder and upper legs were 1.9±0.7 and 2.5±1.1, respectively. Overall, 35% of the stalls were categorized as dirty (>2.5), whereas 13% and 47% of the cows had udder and leg cleanliness scores >2.5, respectively. From the final multivariable models (p<0.05), daily lying time increased by 1.0 h for cows older than 5.25 years versus younger cows. Conversely, lying time decreased by 1.0 h with stall cleanliness scores >2.5 and by 1.6 h with poorly positioned neck rails. In an interaction term, addition of new bedding at least once a day without removing stall manure at least once a day decreased the daily lying time of the cows by 1.5 h, whereas failure to add new bedding at least once a day but removing stall manure at least once a day decreased the lying time of the cows by 1.2 h. Farm-level risk factors for stall dirtiness (>2.5) included delayed cleaning of the alley (odds ratio [OR]=6.6, p=0.032), lack of bedding (OR=4.9, p=0.008), and standing idle and/or backward in the stall (OR=10.5, p=0.002). Stalls categorized as dirty (OR=2.9, p=0.041) and lack of bedding (OR=2.7, p=0.065) were cow- and farm-level risk factors for dirtiness of the udder (>2.5), respectively, whereas the stall being dirty (OR=2.3, p=0.043) was the only risk factor (cow level) for dirtiness of the upper legs (>2.5). CONCLUSION: It was recommended that farmers should pay attention to the specific factors identified regarding the stall design (e.g., neck rail position) and bedding/manure management that impact the cleanliness of cows and their lying time.

Animal and management factors associated with weight gain in dairy calves and heifers on smallholder dairy farms in Kenya.

Calf growth is an important determinant of dairy herd productivity, particularly in countries where the dairy industry is expanding, such as Kenya. Our objective was to determine factors associated with weight gain in randomly selected dairy calves and heifers in smallholder dairy farms (SDF) in Kenya. A cross-sectional study on a census of 321 calves and heifers (dairy calves and heifers up to 36 months of age), sampled from 200 randomly selected SDF in Naari, Kenya, formed the study population. Youngstock management was recorded using a questionnaire. Biodata were obtained through subsequent physical examination and heart girth measurement. Descriptive statistical analyses were conducted, and mixed model regression was used for identification of factors associated (p < 0.05) with the natural log transformation of estimated average daily weight gain (ADG). Median and mean ADG of the youngstock were 360 and 443 (s.d. = 375) g day-1, respectively. In the final model, ADG was highest in pre-weaned calves and declined with age. Supplementing with quality hay during the dry season at least weekly was associated with increased ADG. There was an interaction between breed and historical disease on ADG such that disease was associated with decreased ADG in Bos taurus breeds, while ADG in Bos indicus breeds was not affected by disease. There was a significant interaction between education levels of the husband and wife caretakers; when the man's education was low (having less than or equal to primary school), ADG was highest when the woman had not completed primary school, but was lower when the woman had completed primary, secondary or tertiary education, possibly because such women worked off-farm more often. General growth performance of animals on these farms was lower than benchmarked standards recommended for optimum dairy production but within previously reported ranges for the East African region. Supplementation of diets (with hay and/or concentrates) is recommended for optimum growth in calves and heifers, especially in the dry season. If nutritional management of Bos taurus youngstock is not improved, crosses of Bos indicus could be better suited for the current nutritional management systems in SDF in Meru having calf disease problems. Interventions to support educating women (the primary caretakers of the cattle) and men in the community on calf management should be initiated, preferably with shared on-farm responsibilities. Training in better management, even for learned farmers, would be critical to better calf growth.

Risk factors associated with Cryptosporidia, Eimeria, and diarrhea in smallholder dairy farms in Mukurwe-ini Sub-County, Nyeri County, Kenya.

AIM: This study was undertaken to determine the household, calf management, and calf factors associated with the occurrence of Eimeria, Cryptosporidia, and diarrhea in pre-weaned calves reared in smallholder dairy farms in Mukurwe-ini Sub-County of Nyeri County, Kenya. In addition, the study also evaluated factors associated with average daily weight gain in the same pre-weaned calves. MATERIALS AND METHODS: A total of 112 newborn calves (63 males and 49 females) on 111 farms (1 set of twins) were followed for 2 months between June 2013 and August 2013. Two calves were lost to follow-up. A pre-tested questionnaire was used to collect data on household characteristics and calf management practices in the 111 selected farms. On the first visit to the farm (within 7 days of the birth of the calf), blood samples were collected from the jugular vein to assess the level of maternal immunity acquired by the calf, by determining the serum total protein and selenium concentration. At 4 and 6 weeks of age, fecal samples from the calves were collected to assess the presence of Cryptosporidia and Eimeria oocysts. Every 2 weeks for 2 months, the calves and their environments were examined, their 2-week consumption and health history were recorded, and weights were estimated with a weight tape. Each of the factors was evaluated in a univariable regression model and only those found to be significant (p≤0.20) were included in a multivariable model. Elimination of non-significant factors was done in the multivariable model through a backward elimination procedure so that only those variables which were confounders, and/or significant at (p≤0.05) remained in the final model. RESULTS: About 37% (41/110) of the calves experienced diarrhea at least once during the 2-month study period. The overall period prevalence of Eimeria and Cryptosporidia was 42.7% (47/110) and 13.6% (15/110), respectively. Low serum protein was associated with 1.8 and 2.4 times the odds of Eimeria and Cryptosporidia infections, respectively. Lack of supervision of calf birth and low serum total protein were both associated with 1.3 times the odds of diarrhea incidence. Dirty calf pens, feeding <5 L of milk/day, and infection with Eimeria were associated with 0.105, 0.087, and 0.059 kg, respectively, reduced average daily weight gain of the calves. CONCLUSION: In the Kenyan context, calf diarrhea risk could be reduced through better supervision of parturition and colostrum provision. Specifically, the risk of Eimeria and Cryptosporidia infections could be reduced by optimizing the passive transfer of immunity to the newborn calves. Average weight gains of calves could be improved by good colostrum provision, pen hygiene, and preventing Eimeria infections.

Monensin use against Neospora caninum challenge in dairy cattle.

Using a randomized controlled trial design, a randomly allocated intervention group of 15 cows received a slow-release bolus that delivered 100 days of monensin. The negative control group of 15 cows received a placebo bolus that was identical to the monensin bolus, except without the monensin. Two weeks after bolus administration, all cows were challenged with a 2 ml subcutaneous injection of a live tachyzoite suspension. Whole blood and serum samples were collected from each cow every week for the first month post-challenge, and then every 2 weeks for the next 2 months. The extracted DNA from whole blood was tested for the Nc-5 gene fragment of Neospora caninum using a quantitative real time polymerase chain reaction. Serum was tested for antibodies to N. caninum using the IDEXX ELISA. Cows treated with monensin boluses had a significantly lower humoral immune response than cows treated with placebo boluses at one time point post-challenge (week 4 post-challenge). However, when adjusting for repeated measures within cows, the P value for this humoral difference was 0.098. No DNA for N. caninum was detected in either group, likely due to study design features.

Risk factors associated with Neospora caninum seropositivity in randomly sampled Canadian dairy cows and herds.

Our objective was to determine cow- and herd-level risk factors associated with seropositivity for Neospora caninum in a large number of randomly selected Canadian dairy herds, controlling for important confounding variables and co-infections with bovine leukemia virus (BLV), bovine viral diarrhea virus (BVDV) and Mycobacterium avium subspecies paratuberculosis (MAP). Serum samples were obtained from 30 randomly selected cows, where available, in 240 herds using monthly milk testing, within 6 of 10 provinces, and these samples were tested for antibodies against BLV, MAP and N. caninum using commercially available ELISA test kits. Five unvaccinated cattle >6 months old from each herd were tested for antibodies to BVDV using virus neutralization. Most herd-level predictors were obtained through personal interviews with questionnaires administrated to each farm manager. A mixed logistic-regression model was built using N. caninum serostatus at the cow-level as the outcome variable, with herd as a random effect and province as a fixed effect. A BLV seropositive cow was 1.50 times more likely to be seropositive for N. caninum than a BLV-seronegative cow, and this was the only cow-level variable to remain in the final model. Regarding herd-level variables, with "no on-farm dogs" as the baseline, "presence of dogs but not known to eat placentas and/or fetuses" increased the odds of seropositivity for N. caninum by a factor of 1.66. For "presence of dogs known to eat placentas and/or fetuses", the odds ratio (OR) was 2.75, demonstrating a dose-response relationship. "Using embryo transfer" (OR=0.69), "asking for a BVDV-negative test before introducing an animal" (OR=0.30), "using monensin in dry cows" (OR=0.71), and "heifers having nose-to-nose contact with calves" (OR=0.73) were all dichotomous variables negatively associated with seropositivity for N. caninum. "Number of milk cows on the farm" (OR=0.99), and "area (acres) used for forage production" (OR=0.99) were continuous variables negatively associated with N. caninum seropositivity.

Associations between reproductive performance and seropositivity for bovine leukemia virus, bovine viral-diarrhea virus, Mycobacterium avium subspecies paratuberculosis, and Neospora caninum in Canadian dairy cows.

Our objective was to investigate effects of seropositivity for bovine leukemia virus (BLV), Type 1 bovine viral-diarrhea virus (BVDV), Mycobacterium avium subspecies paratuberculosis (MAP), and Neospora caninum (NC), and their possible interactions, on reproductive efficiency (specifically, first-service conception [FSC], and calving interval [CI]) in dairy cows. The sample population included up to 30 randomly selected animals from 179 randomly selected farms in five provinces in Canada, from which 23 farms did not meet the inclusion criteria for the final dataset. Serum samples were tested for antibodies against the stated pathogens using commercially available diagnostic tests. A Cox proportional hazards model with shared (herd-level) frailty was utilized to analyze the CI data. In this model, BLV-seropositive cows had a 7% lower rate of conception compared to seronegative cows (P=0.06). Mixed logistic regression models of CI>484 days, CI>534 days, and CI>584 days were built to explore factors of long CIs. These cut-offs were selected to represent calving-to-conception intervals of >200 days, >250 days, and >300 days. BLV-seropositive cows had higher odds of having a CI>484 days compared to BLV-seronegative cows, and BLV serostatus interacted with lactation number in this model, with 1st lactation seropositive cows being more likely to have a CI>484 days than older seropositive cows. NC-seropositive cows had a 1.27 times higher odds of exhibiting a CI>484 days, a 1.37 times higher odds of a CI>534 days, and a 1.54 times higher odds of a CI>584 days, compared to NC-seronegative cows. Neither BVDV nor MAP seropositivity showed any significant effect in these models. For the FSC models, a first service was classified successful (pregnancy=1) if it was the cow's last service and she calved 270-290 days later. A mixed logistic regression model of FSC revealed an interaction between NC and BVDV-seropositivity at the herd level, with odds ratios of 0.64, 1.06 and 0.85 for NC-seropositive cows (compared to NC-seronegative cows) in BVDV-seronegative, BVDV-seropositive and BVDV-missing herds, respectively. BLV and MAP seropositivity had no significant impact on FSC. All models controlled for herd-clustering effects, and included parity, linear score of somatic cell counts, peak milk, and province to control for confounding. The overall FSC was 51%, the average CI was 393 days, and 18%, 9% and 5% of lactations had CI>484 days, >534 days, and >584 days, respectively.

Risk factors associated with Mycobacterium avium subspecies paratuberculosis seropositivity in Canadian dairy cows and herds.

Our objective was to determine the risk factors associated with the seroprevalence of Mycobacterium avium subspecies paratuberculosis (MAP) in a large number of randomly selected Canadian dairy herds, controlling for important confounding variables and co-infections with bovine leukemia virus (BLV), bovine viral diarrhea virus (BVDV) and Neospora caninum (NC). Serum samples from 30 randomly selected cows, where available, in 315 herds from seven provinces were tested for antibodies against BLV, MAP and NC using commercially available enzyme-linked immunosorbant assay (ELISA) test kits, while five unvaccinated cattle >6 months old from each herd were tested for antibodies to BVDV. We used a zero-inflated negative-binomial (ZINB) multivariable model to determine simultaneously the risk factors associated with the count of MAP-seropositive cows in a herd, and the odds of herds having no MAP-seropositive cows as compared to having one or more MAP seropositive cows in a herd. The following factors were significantly positively associated with the count of MAP-seropositive cows: "more than one cow in the maternity pen", "group-housing for pre-weaned calves in winter", "open heifers purchased during the last 12 months", "beef cattle direct (nose-to-nose) contact", "BVDV-seropositive herds (> or = 1 animal with > or = 1:64 titer)" and "BVD vaccination not done properly in calves" (i.e. after 6 months old, animals were not boostered 2-4 weeks after their first killed vaccine, or not given modified live vaccine), with count ratios of 1.7, 2.0, 2.3, 1.9, 1.4 and 1.8, respectively. The variable "BVDV vaccination (modified live) done properly in calves" (i.e. received another modified live vaccination after 6 months as well) was associated with 0.4 times fewer MAP-seropositive cows.

Milk urea-nitrogen negatively affected first-service breeding success in commercial dairy cows in Prince Edward Island, Canada.

Our objective of this study was to investigate the relationship between milk urea-nitrogen concentrations ([MUN]) and first-service breeding success (FSBS) in a large number of commercial dairy herds, using various timings on [MUN]. All commercial dairy herds in Prince Edward Island on monthly milk testing (n=198) formed the sampling frame. Milk components, [MUN], 24-h milk production, and breeding data for all cows from these farms were gathered electronically from a central database. A first service between 1 June 1999 and 31 May 2000 was classified successful (FSBS=1) if it was the cow's last service and she calved 270-290 d later. Mixed logistic-regression modeling was used to determine the association between FSBS (the outcome variable) and the [MUN] closest to first service, controlling for other possible confounders and clustering effects of cows within the study herds. The final dataset included 2787 successful and 3015 unsuccessful first services. A change in [MUN] on the test closest to first service from 10 to 20 mg/dL was associated with a 13.9% reduction in the odds of FSBS (controlled for parity, milk production and days in milk).

Production effects of pathogens causing bovine leukosis, bovine viral diarrhea, paratuberculosis, and neosporosis.

The primary purpose of this research was to determine associations among seropositivity for bovine leukemia virus (BLV), bovine viral diarrhea virus (BVDV), Mycobacterium avium ssp. paratuberculosis (MAP), and Neospora caninum (NC) and each of 3 outcome variables (305-d milk, fat, and protein production) in Canadian dairy cattle. Serum samples from up to 30 randomly selected cows from 342 herds on monthly milk testing were tested for antibodies against BLV (IDEXX ELISA; IDEXX Corporation, Westbrook, ME), MAP (IDEXX or Biocor ELISA; Biocor Animal Health, Inc., Omaha, NE), and NC (IDEXX or Biovet ELISA; Biovet Inc., St. Hyacinthe, Quebec, Canada). Up to 5 unvaccinated cattle over 6 mo of age were tested for virus-neutralizing antibodies to the Singer strain of type 1 BVDV. Dairy Herd Improvement records were obtained electronically for all sampled cows. Linear mixed models with herd and cow as random variables were fit, with significant restricted maximum likelihood estimates of outcome effects being obtained, while controlling for potential confounding variables. Bovine leukemia virus seropositivity was not associated with 305-d milk, 305-d fat, or 305-d protein production. Cows in BVDV-seropositive herds (at least one unvaccinated animal with a titer > or =1:64) had reductions in 305-d milk, fat, and protein of 368, 10.2, and 9.5 kg, respectively, compared with cows in BVDV-seronegative herds. Mycobacterium avium ssp. paratuberculosis seropositivity was associated with lower 305-d milk of 212 kg in 4+-lactation cows compared with MAP-seronegative 4+-lactation cows. Neospora caninum seropositivity in primiparous cows was associated with lower 305-d milk, fat, and protein of 158, 5.5, and 3.3 kg, respectively, compared with NC-seronegative primiparous cows. There were no interactions among seropositivity for any of the pathogens and their effects on any of the outcomes examined, although the low MAP seroprevalence limited this analysis. Results from this research will contribute to understanding the economic impacts of these pathogens and justify their control.

Comparison of serological methods for the diagnosis of Neospora caninum infection in cattle.

The aims of this study were to evaluate the performance and agreement of various commercial and in-house Neospora caninum antibody assays used in dairy cattle in North America, and to investigate reproducibility of two assays performed in different laboratories. From 1998 to 2005, three enzyme linked immunosorbent assays (ELISAs, a competitive ELISA-VMRD Inc., an indirect ELISA-Biovet Inc., and another indirect ELISA-Herdchek IDEXX Corp.), two indirect fluorescent antibody tests (IFATs, VMRD Inc., and in-house USDA) and one N. caninum agglutination test (NAT, in-house USDA) were utilized to test 397 randomly selected dairy cattle serum samples from 34 herds in eastern Canada for antibodies to N. caninum. The manufacturers' recommended cut-off values were used to evaluate test performance and agreement between tests. One IFAT (VMRD Inc.) performed well (sensitivity and specificity: 0.97 and 0.97, respectively) using reference sera (n = 452), therefore, results from this IFAT on the 397 samples could subsequently be used as the reference standard to calculate test characteristics for the other assays. Only 11% of the 397 sera were found to be N. caninum-positive with the IFAT. Prevalence-adjusted bias-adjusted kappa (PABAK) ranged from 0.06 to 0.99. Positive agreement was moderate to very good (P(pos) = 0.25-0.96). Negative agreement was very good for all assays (P(neg) > 0.94) except NAT (P(neg) = 0.66). Sensitivity was > or =0.89 for all assays except the NAT, which had a significantly lower sensitivity (0.66). Specificity was high (>0.94) for all assays except for one indirect ELISA (specificity = 0.52). This indirect ELISA did not perform satisfactorily when used in 1998, but an improved version of the ELISA performed as one of the best assays in 2004. Reproducibility of the competitive ELISA was excellent, but the reproducibility of the indirect ELISA that was improved was low (concordance correlation coefficient = 0.90 and 0.36, respectively). The performance characteristics observed for most assays in this study make them useful for screening antibodies to N. caninum in cattle.

Comparison of two enzyme-linked immunosorbent assays for diagnosis of Mycobacterium avium subsp. paratuberculosis.

Enzyme-linked immunosorbent assays (ELISAs) are used in Johne's disease (JD) control programs as a first screening for presence of the disease in a herd. A high sensitivity of the ELISA is therefore important, yet the commonly used ELISAs have relatively low sensitivity. The inclusion of an absorption phase, although improving specificity, potentially decreases sensitivity. Sera and feces of 383 adult dairy cows in 8 herds were used to compare the test characteristics of an absorbed and a nonabsorbed indirect ELISA for the detection of JD. The absorbed ELISA is based on a protoplasmic antigen, whereas the nonabsorbed uses a lipoarabinomannan-based antigen. The potential advantage of the nonabsorbed ELISA is that it may be less specific and more sensitive. Two herds certified free of JD were used to compare the specificity of the ELISAs. The other herds used to compare sensitivity were either infected with Mycobacterium avium subsp. paratuberculosis or had unknown status. Using fecal culture as a gold standard, the diagnostic specificity for the absorbed and nonabsorbed ELISAs were 98.4% and 87.9%, respectively. The diagnostic sensitivity was 72.4% and 65.5% for the absorbed and the nonabsorbed ELISA, respectively. Furthermore, a comparison using a fecal DNA probe as the comparison standard resulted in both ELISAs having a sensitivity of 61.9%. Agreement between the 2 ELISAs was moderate, with a kappa statistic of 0.58. The nonabsorbed ELISA did not have a higher sensitivity and had a lower specificity than the absorbed ELISA. Therefore, in this population, there was no advantage gained with using the nonabsorbed ELISA.

Cow-level prevalence of paratuberculosis in culled dairy cows in Atlantic Canada and Maine.

The prevalence of Mycobacterium avium ssp. paratuberculosis (Mptb) in culled dairy cattle in Eastern Canada and Maine was determined to be 16.1% (95% confidence interval 13.8 to 18.3%) based on a systematic random sample of abattoir cattle. Mesenteric lymph nodes and ileum from 984 cows were examined by histologic and bacteriologic methods. Histological testing was far less sensitive than bacteriologic methods for detecting infected cattle. A seasonal pattern of positive cows was also detected, with the highest proportion of cows being Mptb-positive in June (42.5%). Overall, body condition score was not associated with prevalence of Mptb isolation.

Bulk tank milk urea nitrogen: seasonal patterns and relationship to individual cow milk urea nitrogen values.

The objectives of this study were: 1) to determine if bulk tank milk urea nitrogen (BTMUN) and whole herd weighted average of the individual cow MUN levels (WHMUN) were equivalent measurements of herd MUN status; and 2) to determine the seasonal variation in BTMUN concentrations in Prince Edward Island (PEI) dairy herds. For BTMUN-WHMUN correlation testing, bulk tank milk samples from 176 herds were tested for MUN once every 1 to 2 wk between September 1999 and August 2002, as part of routine BTM testing for milk components. During this 3-year period, all herds had all milking cows tested for MUN once a month at the same lab. The WHMUN levels (weighted for milk production) were calculated for each month, and were compared to BTMUN levels using a concordance correlation coefficient (CCC) and a graphic procedure. Tests were only compared if they occurred on the same date, producing a final dataset of 669 comparisons. The BTMUN had good (but not perfect) correlation with WHMUN (CCC = 0.91). This high reliability extended to both the pasture and non-pasture seasons, various milk sampling protocols, and all herd sizes seen in PEI. For evaluating the seasonal variation of BTMUN, the 3 y worth of data (24 803 observations) were divided into 15 seasonal categories, 5 seasons per year (early, mid, and late pasture, and early and late stable). Using linear mixed modelling, significantly (P < 0.05) higher BTMUN values were found during the mid and late pasture seasons of 2000, likely because the precipitation was unusually high during this period, enhancing pasture growth.

Prevalence estimates for paratuberculosis adjusted for test variability using Bayesian analysis.

The ELISA tests that are available to detect an infection with Mycobacterium avium subsp. paratuberculosis (MAP) have a limited validity expressed as the sensitivity (Se) and specificity (Sp). In many studies, the Se and Sp of the tests are treated as constants and this will result in an underestimation of the variability of the true prevalence (TP). Bayesian inference provided a natural framework for using information on the test variability (i.e., the uncertainty) in the estimates of test Se and Sp when estimating the TP. Data from two prevalence studies for MAP using an ELISA in several regions in two locations were available for the analyses. In location 1, all cattle of at least 3 years of age were sampled in approximately 90 randomly sampled herds in each of the four regions of the country. In location 2, in 30 randomly sampled herds in each of three regions, approximately 30 randomly selected cows were sampled. Information about the unknown test Se and Sp and MAP prevalence was incorporated into a Bayesian model by joint prior probability distributions. Posterior estimates were obtained by combining the actual likelihood with the prior distributions using Bayes' formula. The corrected cow-level TP (proportion of infected cows in a herd) was low, 5.8 and 3.6% in locations 1 and 2, respectively. Certain regions within a location differed significantly in herd-level TP (proportion of infected herds). The herd-level TP was 54.3% in location 1 (95% credible interval (CI) 46.1, 63.3%) and 32.9% in location 2 (95% CI: 14.4, 73.3%). The variation in the herd-level TP estimate for location 2 was more than three times as large as the variation in location 1 mainly because of the relatively small number of investigated herds in location 2. In future prevalence studies for MAP, sample size calculations should be based on a very low cow-level prevalence. Approximately 50 and 90% of the herds in the current study had an estimated cow-level TP below 4 and 10%, respectively.

The effect of non-nutritional factors on milk urea nitrogen levels in dairy cows in Prince Edward Island, Canada.

We determined the effects of non-nutritional factors such as breed, parity, days in milk (DIM), milk production, milk quality and milk components on milk urea nitrogen (MUN) concentration. A total of 177 dairy farms in Prince Edward Island containing 10,688 lactating dairy cows participated in the project. Individual-cow milk samples (n=68,158) were collected monthly from July 1999 to June 2000 from each farm. MUN was measured using a Fossomatic 4000 Milkoscan Analyzer at the PEI Milk Quality Laboratory. Descriptive statistics for MUN, parity, DIM, and test-day milk yield, fat and protein were calculated. Mixed linear-regression models were used; "cow" and "herd" were included as random effects to control for the effect of clustering of MUN test dates within cow, and clustering of cows within herd, respectively. The MUN was lower during the first month of lactation, peaked at 4 months of lactation, and decreased later in lactation. A positive relationship existed between MUN concentration and milk yield, while negative relationships with milk protein% and linear score were observed. A quadratic relationship existed between milk fat% and MUN concentration, with higher MUN occurring at mid-range fat percentages. The variation at the herd and cow levels in the model were 19.7 and 19.0%, respectively; while the variation at the test date level was 61.3%. The non-nutritional factors studied explained 13.3% of the variation in MUN.

Evaluation of the reliability and repeatability of automated milk urea nitrogen testing.

The purpose of this study was to evaluate the reliability (precision and accuracy), and repeatability of an infrared method of determining milk urea nitrogen. The reference method used for the reliability assessment was a wet-chemistry, enzymatic determination of milk urea nitrogen. Reliability and repeatability, as measured by concordance correlation coefficients, were 0.97 and 0.99, respectively.

Seroprevalence of infection with Mycobacterium avium subspecies paratuberculosis, bovine leukemia virus, and bovine viral diarrhea virus in maritime Canada dairy cattle.

The purpose of this study was to survey the seroprevalence of infection with the agents of production-limiting diseases in dairy cattle in New Brunswick, Nova Scotia, and Prince Edward Island. In 30 randomly selected herds per province, 30 cattle per herd were randomly selected and tested for antibodies to bovine leukemia virus (BLV) and Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis), while 5 unvaccinated cattle over 6 months of age were tested for antibodies to bovine viral diarrhea virus (BVDV). For BLV, 20.8% (15.8% to 27.0%) of cows were positive, and 70.0% (60.3% to 79.7%) of herds had at least one positive cow. In BLV-positive herds, the average BLV prevalence was 30.9% (24.8% to 37.2%). For M. paratuberculosis, 2.6% (1.8% to 3.9%) of cows were positive, and 16.7% (8.8% to 24.5%) of herds had at least 2 M. paratuberculosis-positive cows. In M. paratuberculosis-positive herds, the average M. paratuberculosis prevalence was 8.5% (6.9% to 10.1%). For BVDV, 46.1% (35.5% to 56.7%) of herds had at least 1 BVDV-positive animal with a titer greater than or equal to 1:64.