Short communication: Extended-spectrum cephalosporin-resistant Escherichia coli in colostrum from New Brunswick, Canada, dairy cows harbor blaCMY-2 and blaTEM resistance genes.

Dairy calves are colonized shortly after birth by multidrug resistant (MDR) bacteria, including Escherichia coli. The role of dairy colostrum fed to calves as a potential source of MDR bacteria resistance genes has not been investigated. This study determined the recovery rate of extended-spectrum cephalosporin-resistant (ESC-R) E. coli in colostrum from cows. The ESC-R E. coli isolates were further investigated to determine their phenotypic antimicrobial resistance pattern and the genes conferring ESC-R. Fresh colostrum was collected from 452 cows from 8 dairy herds in New Brunswick, Canada. The ESC-R E. coli was isolated from the colostrum by using the VACC agar, a selective media for extended-spectrum ß-lactamase producing Enterobacteriaceae. Minimum inhibitory concentration was determined for all the suspected ESC-R E. coli isolates using a commercial gram-negative broth microdilution method. Two multiplex PCR were conducted on all the suspected ESC-R E. coli isolates to determine the presence of the blaCTX-M (groups 1, 2, 9, and 8/25) blaCMY-2, blaSHV, and blaTEM resistance genes. The ESC-R E. coli were detected in 20 (4.43%) of the colostrum samples. At least 1 ESC-R E. coli isolate was detected in 6 (75%) of the dairy herds. All ESC-R E. coli had MDR profiles based on minimum inhibitory concentration testing. No blaCTX-M groups genes were detected; however, the blaCMY-2 gene was detected in 9 or 20 (45%) and blaTEM was detected in 7 of 20 (35%) of the ESC-R E. coli. No ESC-R E. coli had both blaCMY-2 and blaTEM resistance genes. This is the first report of blaCMY-2 and blaTEM genes found in E. coli isolates cultured from dairy colostrum to our knowledge.

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.

Evaluation of environmental and comfort improvements on affective welfare in heifer calves on smallholder dairy farms.

A controlled trial on zero-grazed smallholder dairy farms was conducted to determine the effect of environmental and comfort improvements on sucking and lying behaviours in heifer calves on Kenyan smallholder dairy farms. The study involved 187 heifer calves from 150 farms in two Kenyan counties, 75 farms per county. Farms in one county received animal welfare training and improvements in the calf pen that included: 1) placement of rubber mats on the lying area; 2) fixing gaps/holes in the flooring and roofing; and 3) attaching a rubber nipple on the wall of the calf pen. During the 16-month data collection period, bimonthly farm visits were used to collect data on lying time (using accelerometers) and other animal- and farm-level factors. Multilevel mixed-effects linear regression was used to model daily lying times and frequency of lying bouts, with the animal as a random effect. Over the visits, daily lying times and lying bout durations averaged 12.6-86.7 min/bout, respectively, while the median for the frequency of lying bouts was between 30-46/day. Provision of rubber nipples for non-nutritive sucking lowered proportions of cross-sucking, self-sucking and object-sucking behaviours slightly but not significantly. In a final daily lying time model, superficial lymph node enlargement, body condition score and use of wood shaving/ sawdust/ crop waste as beddings had positive associations. In contrast, group housing and rubber mat use had negative associations with daily lying time. In an interaction term, lying time was significantly higher for calves on clean versus dirty floors if the age was <190 days but this difference diminished significantly in older animals. In a second interaction term, lying time was lower for calves with leaking versus non-leaking roofs, regardless of the pen floor level, but lying time was higher on elevated than non-elevated floors if the roof was intact. In the final model of the frequency of lying bouts, the use of a rubber mat, the years of experience in dairy farming, and calf body weight had negative associations. In contrast, body condition score had a positive association. In an interaction, the frequency of daily lying bouts was lower on clean floors than dirty floors, irrespective of tethering status, but when the floor was dirty, the lying bouts were higher for animals not tethered than the ones sometimes tethered. We conclude that the comfort improvements enhanced the welfare and lying experience of heifer calves on smallholder dairy farms.

Evaluation of influence of milk urea nitrogen on reproductive performance in smallholder dairy farms.

OBJECTIVE: The aim of this study was to determine the relationship between milk urea nitrogen (MUN) and reproductive performance in dairy cows in western Thailand. DESIGN: All cows calving from November 2014 to April 2015 were included in the study, a total of 486 cows from 47 farms. Each cow had milk constituents and MUN tested monthly up to confirmed conception or until the 8th month after parturition. Each farm had a dietary assessment completed. Cox proportional hazard models with shared frailty were used to determine associations of conception rate. RESULTS: Cows became pregnant increasingly quickly over time, except during 100-150 days of lactation. A change in MUN from 12.5 to 13.5 mg/dL on the closet day to breeding was associated with a 5% decrease in conception. Milk protein was negatively associated with hazard of conception, whereas milk lactose and dietary protein:energy ratio had positive associations with conception rate. Breeding season was also significant; the highest conception rate was observed in cows inseminated during winter, whereas insemination during the humid rainy season resulted in the lowest conception rates. The farm random effect in the model was strongly significant. CONCLUSION: Detrimental effects of higher MUN on rate of conception were identified. The rate of conception was positively associated with protein:energy ratio in the study. Therefore, good nutritional management leading to positive energy balance should benefit conception rates.

Management factors associated with time-to-onset and fecal egg/oocyst counts of gastrointestinal parasites in heifer calves on Kenyan smallholder farms.

A prospective cohort study on zero-grazed smallholder dairy farms was conducted to determine factors associated with onset and counts of gastrointestinal parasitism in heifer calves. The researchers recruited 187 newborn heifer calves from 150 farms in Kenya. Over 16 months, farm visits every two months were used to collect rectal fecal samples and animal- and farm-level measures. Fecal samples underwent centrifugal fecal flotation with Sheather's sugar to determine counts of strongyle-type eggs and coccidia oocysts. Cox proportional hazard (Cox PH) analysis and mixed-effects negative binomial (MeNB) regression determined factors associated with time-to-onset and counts of strongyle-type eggs and coccidia oocysts, respectively (P < 0.05). The incidence rate of strongyles was 0.0011 cases/animal-day while coccidia was 0.0073 cases /animal-day. Incidence risks of strongyles and coccidia over the study period were 28.3 % (53/187) and 87.7 % (164/187), respectively. For infected calves, median time-to-onset for strongyles and coccidia was 78 (interquartile-range: IQR 38-117) and 43 (IQR 29-92) days, respectively. In the final Cox PH model for strongyles, breed (Ayrshires and Jerseys) and weaned calves had a greater hazard of infection than Friesians and preweaned calves, respectively. Calf tethering outside the pen was associated with a higher hazard of strongyle infection. In the final Cox PH model for coccidia, calves with watery and/or hemorrhagic diarrhea had a higher hazard compared with those with hard or soft feces. Weaning status and birth weight (kg) were time-varying covariates, leading to increased hazard over time. In the final MeNB model for strongyles, weaned animals had higher counts than those still on milk. In an interaction variable, the predicted strongyle-type egg counts increased with longer duration of farm operation when herd size was less than five cattle, but decreased when herd size was more than five. In the final MeNB model for coccidia, calves sometimes tethered outside their pens had higher counts than those continuously enclosed in the pen. Calf pen floors with either scant manure or moderate slurry had higher predicted counts than those on a clean pen floor. Calves with watery or hemorrhagic diarrhea and fed fresh Napier grass (Pennisetum purpureum) had higher counts compared with soft or hard feces and those not given fresh Napier, respectively. In an interaction variable, calves experiencing diarrhea and raised on elevated slatted floors had lower oocyst counts compared with those having diarrhea but not on elevated floors. The identified management practices associated with onset and counts of gastrointestinal parasitism should be considered in control efforts.

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.

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.

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.

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.

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.

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.

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.

Assessment of Effectiveness of Control Strategies Against Simulated Outbreaks of Highly Pathogenic Avian Influenza in Ontario, Canada.

The North American Animal Disease Spread Model (NAADSM) is a stochastic model framework developed to simulate the spread of highly contagious diseases of livestock and poultry, such as foot-and-mouth disease and highly pathogenic avian influenza (HPAI). The objective of this study was to make recommendations on the most effective HPAI control policy for Canada, specifically, on the effect of different speeds of detection, effectiveness of movement restrictions and stamping-out and ring-culling strategies on the magnitude of an HPAI outbreak. In addition, the effect of introduction of infection in a range of multiple farms simultaneously was also evaluated. A total of 21 060 scenarios, defined as different combinations of parameters for various epidemiological conditions and control measures, were created to simulate the number of poultry flocks that would become infected as a result of an incursion of HPAI. Each scenario was parameterized in NAADSM and replicated 1000 times, generating the median number of flocks infected at the end of the simulated outbreak for each scenario. Negative binomial regression analysis was used to model significant explanatory variables of the median number of flocks infected at the end of each simulated outbreak for each of the 21 060 scenarios. The final model included the following explanatory variables: number and type initially infected flock(s), density of flocks within the county where the initially infected flock(s) was located, probability of transmission through indirect contact, subclinical spread of the infection, speed of detection and a two-way interaction between intensity of bird destruction strategy and movement restriction effectiveness to reduce transmission through direct and indirect contacts. The modelling results suggested that stamping out of the detected infected flocks, without ring culling, in combination with effective movement restrictions on direct and indirect contacts, would be the most appropriate policy for Ontario.

Identification of effective treatment criteria for use in targeted selective treatment programs to control haemonchosis in periparturient ewes in Ontario, Canada.

Haemonchosis is often associated with late gestation and parturition in ewes in Canada. Due to widespread concerns about development of anthelmintic resistance (AR), targeted selective treatment (TST), where individual animals are treated with an anthelmintic rather than the entire flock, is a possible strategy to control clinical signs in recently lambed ewes while still maintaining parasite refugia. Performing fecal egg counts (FEC) on individual animals is often cost-prohibitive, so indicators that identify ewes with high FEC are essential for TST programs. The study objectives were to: a) evaluate the ability of four TST indicators to identify periparturient ewes with high Haemonchus sp. FEC and b) determine appropriate treatment thresholds for statistically-significant indicators. A field study was conducted during the 2013 and 2014 lambing seasons (February-May) on three client-owned farms in Ontario with documented AR and problems with haemonchosis in ewes. Ewes were examined within three days of lambing and selected for treatment with oral closantel (10mg/kg body weight), a novel anthelmintic to Canada, if they met at least one of four criteria: a) the last grazing season was their first grazing season; b) body condition score ≤2; c) Faffa Malan Chart (FAMACHA©) score ≥3; and/or d) three or more nursing lambs. Fecal samples were collected per rectum on the treatment day from each of 20 randomly selected treated and untreated ewes on each farm. Haemonchus sp. percentages on each farm, as determined by coproculture, ranged from 53% to 92% of total fecal trichostrongyle-type egg counts. Mean Haemonchus sp. FECs were significantly higher in treated ewes (n=136) than in untreated ewes (n=103) on the day of treatment in both years (p=0.001), suggesting the indicators were suitable for identifying animals with high Haemonchus sp. FEC. A linear mixed model was fit with logarithmic-transformed Haemonchus sp. FEC as the outcome variable, the four indicators and year as fixed effects, and farm as a random effect. FAMACHA© score was the sole indicator to remain significantly associated with FEC (p=0.002). A receiver-operator curve determined that test sensitivity was maximized (92.4%) with FAMACHA© score ≥3 as the sole indicator. FAMACHA© score should therefore be included in TST programs to identify ewes requiring treatment at lambing due to Haemonchus sp.

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.

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.

Use of in-stream reservoirs to reduce bacterial contamination of rural watersheds.

An investigation into bacterial water quality problems was conducted on an interconnected stream and irrigation system within the Oldman River Basin of southern Alberta, Canada. Levels of indicator bacteria, including fecal coliforms, generic Escherichia coli and fecal streptococci, were repeatedly measured in streams and irrigation return canals of this river basin during the summer of 2001. Bacterial-loading segments of the irrigation/stream system were identified through a comparison of indicator bacteria levels in pairs of upstream and downstream sites. Mann-Whitney U-tests indicated that reservoirs significantly reduced bacterial counts. A temporal comparison of E. coli counts and river discharges suggested that these indicator bacteria do not originate from within in-stream sediments. Site-specific as well as cumulative inputs from a variety of non-point sources are likely to be responsible for the high downstream levels of indicator bacteria in this water system. The use of management practices such as in-stream reservoirs may significantly reduce contamination, and increase the quality of limited rural water supplies to allow their reuse and safe discharge into downstream water sources. The identification of bacteria-loading river/canal segments could also be used to prioritize restoration projects.

Cow, farm, and ecologic risk factors of clinical vesicular stomatitis on Costa Rican dairy farms.

Twenty-two Costa Rican dairy herds were actively monitored for vesicular diseases between November 1991 and March 1992, in anticipation of the annual dry season occurrence of vesicular stomatitis (VS). Thirty-nine confirmed cases of VS were concentrated on nine farms, resulting in an average herd incidence rate of 11.1% of mature cattle for the affected farms, or 2.6% for all farms. Affected cattle were generally older (average age = 5.4 years), with seven-year-old cows having the highest age-specific incidence rate (6.5%). The New Jersey serotype of VS was diagnosed for all but four cases, primarily manifesting as teat vesicles. Cases had high acute and convalescent antibody titers for both major VS serotypes, New Jersey and Indiana, with significant titer increases after clinical disease. Screening and multiple variable regression analyses of 140 cow, farm, and ecologic variables revealed that clinical disease was significantly associated with cows being in lactation, and with higher acute antibody titers to serotype Indiana, but not with any predisposing diseases. Significant farm factors were the presence of poultry and a longer calving interval on the farm, but not higher rates of other diseases. According to a prior epidemiologic hypothesis, two ecologic factors were forced to be included into the farm model: the reported presence of sand flies, and a higher proportion of the farm being in forest land, with subsequent stepwise regression. The resulting model containing only the two forced variables was significant (P < 0.003) and correctly classified 78% of farms into the correct group, as compared with 79% for the final stepwise model.(ABSTRACT TRUNCATED AT 250 WORDS)

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.