Comparison of calculation methods used for the determination of anthelmintic resistance in sheep in a temperate continental climate.

This study compared results obtained with five different fecal egg count reduction (FECR) calculation methods for defining resistance to ivermectin, fenbendazole, and levamisole in gastrointestinal nematodes of sheep in a temperate continental climate: FECR1 and FECR2 used pre-and posttreatment fecal egg count (FEC) means from both treated and control animals, but FECR1 used arithmetic means, whereas FECR2 used geometric means; FECR3 used arithmetic means for pre- and posttreatment FECs from treated animals only; FECR4 was calculated using only arithmetic means for posttreatment FECs from treated and control animals; and FECR5 was calculated using mean FEC estimates from a general linear mixed model. The classification of farm anthelmintic resistance (AR) status varied, depending on which FECR calculation method was used and whether a bias correction term (BCT, i.e., half the minimum detection limit) was added to the zeroes or not. Overall, agreement between all methods was higher when a BCT was used, particularly when levels of resistance were low. FECR4 showed the highest agreement with all the other FECR methods. We therefore recommend that small ruminant clinicians use the FECR4 formula with a BCT for AR determination, as this would reduce the cost of the FECRT, while still minimizing bias and allowing for comparisons between different farms. For researchers, we recommend the use of FECR1 or FECR2, as the inclusion of both pre- and posttreatment FECs and use of randomly allocated animals in treatment and control groups makes these methods mathematically more likely to estimate the true anthelmintic efficacy.

Comparison of calculation methods used for the determination of anthelmintic resistance in sheep in a temperate continental climate.

This study compared results obtained with five different fecal egg count reduction (FECR) calculation methods for defining resistance to ivermectin, fenbendazole, and levamisole in gastrointestinal nematodes of sheep in a temperate continental climate: FECR1 and FECR2 used pre- and posttreatment fecal egg count (FEC) means from both treated and control animals, but FECR1 used arithmetic means, whereas FECR2 used geometric means; FECR3 used arithmetic means for pre- and posttreatment FECs from treated animals only; FECR4 was calculated using only arithmetic means for posttreatment FECs from treated and control animals; and FECR5 was calculated using mean FEC estimates from a general linear mixed model. The classification of farm anthelmintic resistance (AR) status varied, depending on which FECR calculation method was used and whether a bias correction term (BCT, i.e., half the minimum detection limit) was added to the zeroes or not. Overall, agreement between all methods was higher when a BCT was used, particularly when levels of resistance were low. FECR4 showed the highest agreement with all the other FECR methods. We therefore recommend that small ruminant clinicians use the FECR4 formula with a BCT for AR determination, as this would reduce the cost of the FECRT, while still minimizing bias and allowing for comparisons between different farms. For researchers, we recommend the use of FECR1 or FECR2, as the inclusion of both pre- and posttreatment FECs and use of randomly allocated animals in treatment and control groups makes these methods mathematically more likely to estimate the true anthelmintic efficacy.

Performance of a whole blood interferon-gamma assay for detection and eradication of caseous lymphadenitis in sheep.

Caseous lymphadenitis (CLA), a chronic bacterial disease of sheep and goats caused by Corynebacterium pseudotuberculosis, could be controlled by eradication of infected carriers. This study aimed at validation of a whole blood interferon-gamma (IFN-gamma) enzyme immunoassay (EIA) (Bovigam, Pfizer) in naturally infected sheep for use in eradication of infection from a flock. This assay used formalin-inactivated whole bacterial cells as antigen. The sensitivity of the whole cell assay was improved by increasing both the volume of blood and the number of bacterial cells. The assay was validated in experimentally infected sheep and in a flock of known-negative sheep, as well as in a naturally infected flock, a proportion of which was vaccinated with a commercial CLA vaccine. An optical density (540nm) (OD) cut-off of 0.09 was effective in classifying animals as test positive or negative in the naturally infected flock, although there was variation in OD between visits, notably with weakly reacting animals. The test had a sensitivity of 91% and a specificity of 98%. Postmortem data supported the results in test-negative animals. Visit-to-visit variation in IFN-gamma EIA OD in the naturally infected flock as well as CLA disease status was used to develop an algorithm for the eradication of CLA from a known infected flock. The whole blood IFN-gamma assay shows promise for eradication of caseous lymphadenitis from sheep flocks.

An interferon-gamma assay for diagnosis of Corynebacterium pseudotuberculosis infection in adult sheep from a research flock.

The optimal method of control of caseous lymphadenitis of sheep caused by Corynebacterium pseudotuberculosis is eradication of infection by identification and removal of infected carrier animals. Current serological approaches to identification of infected sheep are generally hampered by low sensitivity and specificity of available tests. The objective of this study was to develop a whole blood assay for detection of C. pseudotuberculosis-infected sheep, based on detection of IFN-gamma response to whole cell C. pseudotuberculosis antigens, and to determine the reliability of the assay. A commercially available bovine interferon-gamma assay enzyme-linked immunosorbent assay was used and the test optimised using experimentally infected sheep. The assay was also tested on known CLA-negative sheep. Setting a IFN-gamma optical density cut-off at 0.100 as positive under the conditions used, the test detected C. pseudotuberculosis experimentally infected sheep over a 450-day period with a reliability of 95.7%. It identified known non-infected sheep with a reliability of 95.5%. Repeated vaccination of three uninfected sheep with a commercially available bacterin-toxoid vaccine did not interfere with the assay. The IFN-gamma response of sheep whole blood to C. pseudotuberculosis antigens offers promise for use in a test-and-removal approach to eradication of CLA in sheep.

Comparison of an interferon-gamma to a phospholipase D enzyme-linked immunosorbent assay for diagnosis of Corynebacterium pseudotuberculosis infection in experimentally infected goats.

The optimal method of control of caseous lymphadenitis of goats caused by Corynebacterium pseudotuberculosis is eradication of infection by identification and removal of infected carrier animals. The objective of this study was to compare detection of C. pseudotuberculosis experimentally infected goats using a commercially available bovine interferon-gamma (IFN-gamma) whole blood enzyme-linked immunosorbent assay (ELISA) to serological response to a recombinant phospholipase D (PLD) ELISA. The tests were assessed repeatedly over 1 year in three infected and three non-infected goats. Using a IFN-gamma optical density cut-off at 0.10 as positive under the conditions used, the test accurately detected C. pseudotuberculosis experimentally infected goats over a 363 day period with a reliability of 89.2% and non-infected goats with a reliability of 97.1%. Using a cut-off value of the mean for negative samples plus two standard deviations, the PLD ELISA detected C. pseudotuberculosis experimentally infected goats over this period with a reliability of 81.0% and non-infected goats with a reliability of 97.0%. The PLD ELISA was however more predictive than the IFN-gamma ELISA of the presence of lesions observed at postmortem examination of infected goats.

Analysis of the immunodominant antigens of Corynebacterium pseudotuberculosis.

Antibodies to seven antigens in a whole cell lysate of Corynebacterium pseudotuberculosis ranging in molecular mass from 22 to 120 kilodaltons (kDa) were present in sera of 40 sheep and goats infected with C. pseudotuberculosis. Three antigens of about 120, 68, and 31.5 kDa in size were consistently detected with sera from all animals and twenty-two sera had antibodies to 64, 43, 40, and 22 kDa antigens. None of these antigens were detected by sera from 160 sheep in a C. pseudotuberculosis-free research flock. An NaCl extract of C. pseudotuberculosis cells contained one major protein of about 31.5 kDa and four minor proteins of 68, 64, 43, and 22 kDa in molecular mass as shown by Coomassie Blue staining. Immunoblot analysis demonstrated that the three immunodominant antigens identified in the whole cell extract were contained in the NaCl extract. The 31.5-kDa protein was purified from the NaCl extract by fast-protein liquid chromatography gel filtration to near homogeneity. The purified 31.5-kDa protein showed phospholipase D activity as indicated by synergistic hemolysis with Rhodococcus equi factors and sphingomyelinase activity. The 31.5-kDa protein reacted with antibodies in serum from a sheep naturally infected with C. pseudotuberculosis. This serum also had phospholipase D neutralizing activity. On the basis of its molecular mass, biological activity, N-terminal amino acid sequence analysis, and immunoreactivity, the 31.5-kDa protein was identified as the phospholipase D exotoxin of C. pseudotuberculosis.

The use of a microagglutination assay for the detection of antibodies to Corynebacterium pseudotuberculosis in naturally infected sheep and goat flocks.

Two goat flocks comprising 326 animals and four sheep flocks comprising 343 animals, all with a previously recognized problem of abscesses due to Corynebacterium pseudotuberculosis, were examined for the presence of abscesses and antibody titers to C. pseudotuberculosis as detected by direct microagglutination assay. In sheep there was a strong positive relationship between age and titer (p less than 0.0001). However, the relationship in goats between age and titer could not be determined due to a strong interaction between flock and age. When the relationship between abscesses and titer was examined, it was found that goats with abscesses had higher titers than those that did not (p less than 0.05), whereas there was no difference in titer between sheep with abscesses and those without (p = 0.5753). The sensitivity of the microagglutination test was poor to good for both species (52.3% for goats and 89.7% for sheep). The specificity of the test was fair to poor (64.9% for goats and 21.7% for sheep). Given a disease prevalence of 13.5% for goats and 8.5% for sheep the predictive value of the positive test was very poor (18.9% for goats and 9.6% for sheep) but the predictive value of the negative test was good to excellent (89.7% for goats and 95.8% for sheep). The poor specificity of the test and therefore the positive predictive value may be due in part to the criterion of classification of presence of disease, i.e. presence of an abscess at the time of sampling.(ABSTRACT TRUNCATED AT 250 WORDS)

A field trial to evaluate a whole cell vaccine for the prevention of caseous lymphadenitis in sheep and goat flocks.

A field trial to evaluate a whole cell vaccine for the prevention of caseous lymphadenitis (CLA) in sheep and goats was performed in one goat herd and one sheep flock over a period of three years. In goats, there was a nonstatistically significant trend for fewer cases of CLA in the vaccinated animals compared to the controls. In sheep, from six months to 36 months postinitial vaccination, the proportion of vaccinated sheep that developed CLA was significantly less (p less than 0.05) than in the control sheep. The antibody titers to Corynebacterium pseudotuberculosis as detected by microagglutination assay were significantly different (p less than 0.0001) at all times except at the initial vaccination. Swellings occurred at the vaccination site at an incidence level of 29.6% in goats and 34.1% in sheep. The vaccine appeared to be efficacious in reducing the proportion of sheep that developed CLA when challenged naturally in a field situation.

Naturally occurring lesions of the uterine tube in sheep and serologic evidence of exposure to Chlamydophila abortus.

The uterine tubes from 405 ewes, collected at an abattoir, were assessed grossly and microscopically for abnormalities that correlated with serological evidence of exposure to Chlamydophila abortus. Gross lesions were found in 41 ewes and 86 had microscopic lesions. Enzyme immunoassay (EIA) of serum was used as an indication of exposure of individual ewes to C. abortus; 52 were found to be positive. Chi-squared analysis indicated no association between EIA-positive animals and lesions of the uterine tube.

Border disease in a flock of sheep: epidemiologic, laboratory, and clinical findings.

A flock of sheep in which border disease (BD) was enzootic was studied through a breeding season. At the beginning of the study (August 1981), 125 (82%) of 152 ewes were seropositive to the cross-reacting bovine viral diarrhea virus. Within 7 months, 3 (18%) of 17 seropositive ewes retested had reverted to seronegative. Of the remaining 21 ewes identified as seronegative, 7 (33%) converted to seropositive by the end of the study. Triplet lambs were born, 2 of which exhibited clinical signs of BD. The virus was isolated from blood lymphocytes from both of the affected lambs. The most severely affected lamb shed virus into the urine, saliva, and feces through 10 weeks of age. Lymphocyte stimulation tests indicated that the lymphocytes from the affected lambs had decreased function in months 4 through 7, but returned to normal function by the eighth month. Transmission of BD virus was investigated by exposing 5 seronegative ewes to the BD-infected lambs. Two of the contact ewes developed viremia and 3 converted to seropositive within the 13-week exposure period. Evidence from this and other studies supports a model of BD in gravid, nongravid, and persistently infected adult sheep.

Mastitis of sheep and goats.

This article presents an overview of the recent research into ovine and caprine mastitis. The common clinical presentations of mastitis in these species are reviewed, as are the important etiologic agents and their significance. The interpretation of somatic cell counts and surrogate tests, factors that affect somatic cell count levels, and association of somatic cell count levels with productivity are reviewed. Investigations into the treatment and prevention of mastitis and milking equipment function are discussed, and comments are made on the public health implications of extra label drug use and the consumption of unpasteurized milk.

Treatment of Goats Infected with the Lungworm Muellerius capillaris.

Goats naturally infected with Muellerius capillaris were treated with ivermectin subcutaneously once or twice at the rate of 200 or 300 mug/kg body wt or with fenbendazole per os twice at 15 mug/kg body wt. Goats ceased passing larvae 11 to 20 days after treatment, and except for one doe, larvae reappeared in feces 34 to 59 days after treatment. In sections of lung of ivermectin-treated goats, adult Muellerius had swollen body walls and disrupted intestinal tracts. Granulomas, some mineralized, were present. It is suggested that immature Muellerius were not destroyed by either anthelmintic and that following destruction of the adults, immature Muellerius resumed development to the adult stage and produced more first-stage larvae. Treatment of Muellerius may be more effective if repeated after approximately a 35-day interval.

An evaluation of antimicrobial therapy for undifferentiated bovine respiratory disease.

A field trial of antimicrobial therapy for cases of undifferentiated bovine respiratory disease (UBRD) in beef calves was conducted at four Ontario feedlots. The primary purpose of the trial was to evaluate the efficacy of three different antimicrobials (oxytetracycline, penicillin, and trimethoprim-sulfadoxine) in the treatment of UBRD occurring within the first 28 days postarrival.The response, relapse, and case fatality rates overall were 85.7%, 14.8%, and 1.4%, respectively, and were not significantly different among the three antimicrobials evaluated. Weight gains of calves treated with the different drugs were not statistically different over the feeding period. Calves that suffered a relapse posttreatment were first treated significantly earlier (p<0.001) in the postarrival period than those that did not relapse. Considered together, treated calves gained significantly less (p<0.05) over the first 28 days and throughout the entire feeding period than controls that were never sick. Cases of UBRD that responded to therapy and did not relapse had rates of gain that were not significantly different from the controls.

An assessment of the utility of microcomputers and dairy herd management software for dairy farms and dairy practices.

Microcomputer systems were placed in three veterinary practices each serving three farms (bureau), and onto nine dairy farms (on-farm). Over a twenty-four week period, the utilization of the computer system and the DHM software was monitored. The on-farm system was more costly in terms of equipment and technical support effort, but the information was utilized to a greater extent than it was by the bureau participants who had invested more user time per cow. Note that actual time will vary with the software program used. The farmers indicated that they wished to have access to the information offered by the software. ;Computer phobia' was not found to be a problem. The expectations of the computer system and its benefits generally remained high. In general, the information available through the microcomputer system and the DHM software was found to be useful to both the dairy farmers and the veterinarians in this study.

Attitudes and expectations of producers to the use of a microcomputer-based management information system to monitor dairy herd performance.

The attitudes and expectations of producers toward the use of a microcomputer-based herd management information system were assessed. The study was conducted over a two-year period, beginning in January 1986, and was operated as a bureau service. The implementation and use of the program are described elsewhere. Pre- and posttrial questionnaires were administered to assess producer attitudes. We found that the monthly analysis reports were used in the management of the dairy farms and were found to be a useful management tool. The majority of producers indicated a willingness to pay, on average, $6.86/cow/year for such a service.

The seroprevalence of maedi-visna in Ontario sheep flocks and its relationship to flock demographics and management practices.

The objectives of this study were to describe the serological prevalence of maedi-visna in a sample of Ontario sheep flocks, and to identify management and demographic variables that were associated with seroprevalence for maedi-visna. A sample of 103 sheep flocks in Ontario was randomly selected from those flocks participating in the Red Meat Plan. The owners of these flocks were surveyed regarding management procedures on their farms, and blood samples were taken from a random sample of ewes in each flock. At least one ewe tested serologically positive, based on the agar gel immuno-diffusion test, in 69.9% of the farms. Positive serological reactions occurred in 20.9% of the 3880 sheep tested. Flock demographics and farm management variables were considered in a multiple regression model, and several factors were positively associated with higher maedi-visna seroprevalence rates. These included the average age of the flock, the number of years the owner had been sheep farming, the practice of using foster ewes, the practice of allowing lambs to have contact with other ewes that are lambing, and the average pasture acreage per ewe.

Implementation and use of a microcomputer-based management information system to monitor dairy herd performance.

A microcomputer-based herd management information system was implemented as part of the herd health program provided to 13 dairy clients by the Ontario Veterinary College, University of Guelph. The study was conducted over a two year period. Data were collected from on-farm event diaries, veterinary visit reports, and production testing information. Selected indices of reproduction, udder health, production, and heifer performance were reported. It was concluded that the implementation of a microcomputer-based information management system, operated as a bureau service, was feasible. However, limitations to the implementation in veterinary practice were identified.

Verminous Pneumonia in Adult Dairy Cows in Southern Ontario due to Dictyocaulus viviparus.

An outbreak of verminous pneumonia due to Dictyocaulus viviparus in a herd of mature, lactating, dairy cows in southern Ontario is described. The fact that the herd had been closed to new additions for ten years and had never experienced clinical disease due to D. viviparus in the past makes the occurrence of this herd problem difficult to explain. Correlation of fecal Baermann analysis for D. viviparus larvae with the progress of anthelmintic treatment is discussed. It is suggested that certain climatological variations in combination with unique, immunological aspects of D. viviparus infection may have contributed to the development of clinical disease in this herd.

Taenia ovis infection and its control: a Canadian perspective.

Distributed worldwide, Taenia ovis infection is responsible for the condemnation of sheep carcasses in many countries. This review highlights the programme used in New Zealand to successfully control T. ovis in sheep, and discusses how similar approaches may be modified for use in Canada, given what is currently known about the epidemiology of T. ovis. The lifecycle of the parasite is well known, involving dogs as the definitive host and sheep or goats as the intermediate host. An effective vaccine does exist, although it is not presently commercially available. In New Zealand an industry-based, non-regulatory programme was created to educate producers about T. ovis and necessary control strategies, including the need to treat farm dogs with cestocides regularly. This programme resulted in a substantial decrease in the prevalence of T. ovis infections between 1991 and 2012. Historically, T. ovis was not a concern for the Canadian sheep industry, but more recently the percentage of lamb condemnations due to T. ovis has increased from 1.5% in 2006 to 55% in 2012. It has been suggested that coyotes may be transmitting T. ovis, but this has not been confirmed. Recommendation are made for the Canadian sheep industry to adopt a control programme similar to that used in New Zealand in order to reduce the prevalence of T. ovis infection.

A systematic review and meta-analysis of factors associated with anthelmintic resistance in sheep.

BACKGROUND: Anthelmintic drugs have been widely used in sheep as a cost-effective means for gastro-intestinal nematode (GIN) control. However, growing anthelmintic resistance (AHR) has created a compelling need to identify evidence-based management recommendations that reduce the risk of further development and impact of AHR. OBJECTIVE: To identify, critically assess, and synthesize available data from primary research on factors associated with AHR in sheep. METHODS: Publications reporting original observational or experimental research on selected factors associated with AHR in sheep GINs and published after 1974, were identified through two processes. Three electronic databases (PubMed, Agricola, CAB) and Web of Science (a collection of databases) were searched for potentially relevant publications. Additional publications were identified through consultation with experts, manual search of references of included publications and conference proceedings, and information solicited from small ruminant practitioner list-serves. Two independent investigators screened abstracts for relevance. Relevant publications were assessed for risk of systematic bias. Where sufficient data were available, random-effects Meta-Analyses (MAs) were performed to estimate the pooled Odds Ratio (OR) and 95% Confidence Intervals (CIs) of AHR for factors reported in ≥2 publications. RESULTS: Of the 1712 abstracts screened for eligibility, 131 were deemed relevant for full publication review. Thirty publications describing 25 individual studies (15 observational studies, 7 challenge trials, and 3 controlled trials) were included in the qualitative synthesis and assessed for systematic bias. Unclear (i.e. not reported, or unable to assess) or high risk of selection bias and confounding bias was found in 93% (14/15) and 60% (9/15) of the observational studies, respectively, while unclear risk of selection bias was identified in all of the trials. Ten independent studies were included in the quantitative synthesis, and MAs were performed for five factors. Only high frequency of treatment was a significant risk factor (OR=4.39; 95% CI=1.59, 12.14), while the remaining 4 variables were marginally significant: mixed-species grazing (OR=1.63; 95% CI=0.66, 4.07); flock size (OR=1.02; 95% CI=0.97, 1.07); use of long-acting drug formulations (OR=2.85; 95% CI=0.79, 10.24); and drench-and-shift pasture management (OR=4.08; 95% CI=0.75, 22.16). CONCLUSIONS: While there is abundant literature on the topic of AHR in sheep GINs, few studies have explicitly investigated the association between putative risk or protective factors and AHR. Consequently, several of the current recommendations on parasite management are not evidence-based. Moreover, many of the studies included in this review had a high or unclear risk of systematic bias, highlighting the need to improve study design and/or reporting of future research carried out in this field.