John R. Campbell

A longitudinal study of gastrointestinal parasites in Canadian dairy farms: The value of an indirect Ostertagia ostertagi ELISA as a monitoring tool

The general objective of this study was to evaluate a crude Ostertagia ostertagi antigen enzyme-linked immunosorbent assay (ELISA) for monitoring gastrointestinal parasites in lactating dairy cattle. A longitudinal study of gastrointestinal parasites in lactating dairy cows was carried out in 38 herds in four provinces of Canada (Prince Edward Island, Quebec, Ontario and Saskatchewan) from September 1999 to October 2000. Bulk tank milk, cow milk, serum and fecal samples were collected monthly or quarterly from all these farms. Information on herd management factors was collected by a standard questionnaire and individual cow production data were obtained from an electronic database. The overall mean optical density ratio (ODR) was 0.30 and ranged from -0.05 to 1.55. Although a clear seasonal pattern was not observed, the ODR values tended to decrease during the housing period and start increasing in the spring before the cows went out to pasture. The second and third or greater lactation cows had significantly higher ODR values compared with first lactation animals. The individual cow ODR had a very low correlation with individual squared root fecal egg counts but showed a reasonably high correlation when herd averages values were computed (r=0.73). A moderate correlation (r approximately 0.50) between the bulk tank and herd average ODR was observed. Milk yield was negatively associated with individual cow milk ODR and a quadratic effect on ODR was observed for days in milk. Twenty-eight of the herds participated in a clinical trial of eprinomectin (Ivomec Eprinex) treatment at calving. The cow level ODR values determined late in the previous lactation had a marginally significant effect (P=0.07) on treatment response, suggesting that high OD cows responded better to the anthelmintic treatment. However, because of the small sample size available in this model, more research is needed to better understand this relationship. In conclusion, the indirect ELISA using an O. ostertagi crude antigen appears useful as a technique for monitoring gastrointestinal parasite burdens in adult dairy cows and holds promise as a potential predictor of response to anthelmintic treatment.

Effect of bovine viral diarrhea virus in the feedlot

It could be argued that bovine viral diarrhea virus (BVDV) is one of the most economically significant infectious pathogens of feedlot cattle. Although the direct economic losses caused by this virus have not been well quantified, the role it plays as an immunosuppressive agent and as a potentiator for other diseases, most notably bovine respiratory disease, have been well documented. It is also a difficult disease for the feedlot veterinarian to control effectively. Individual cattle persistently infected with BVDV often serve as the source of infectious virus within a group of feedlot cattle, and the ultimate responsibility for preventing persistent infections in cattle rests with the cow-calf producer and not with the feedlot owner. The enormous impact of the virus on the livestock industry has led the Academy of Veterinary Consultants to draft a position statement that resolves that the beef and dairy industries adopt measures to control and target eventual eradication of BVDV from North America.

Comparison of the Diagnostic Sensitivity of a Commercially Available Culture Kit and a Diagnostic Culture Test Using Diamond's Media for Diagnosing Tritrichomonas Foetus in Bulls

A number of different culture media have been described for use in the diagnosis of Tritrichomonas foetus infection in bulls, and recently, a commercial culture kit has become available. The objective of this study was to compare the sensitivity of 2 culture-based diagnostic tests for T. foetus in bulls. One test used a commercial kit for transport and culture of the samples. The other test used a thioglycollate transport medium (TFTM) for transport and a modified Diamonds medium (MDM) for culture of the samples. Twenty-one bulls infected with T. foetus were sampled repeatedly. On each sampling day, samples collected from the left and right sides of the bull were tested with one of the 2 diagnostic tests being compared. The effect of the type of diagnostic test on the outcome of the test was evaluated with a chi-square test for the calculated odds ratio. Because repeated tests from the same bull cannot be considered independent measures, unadjusted chi-square tests were adjusted for the effect of clustering by bull. Samples tested using the commercial kit were 6.95 times as likely to be positive as samples tested with a diagnostic test using MDM (P < 0.001).

Effects of temperature on the survival of Tritrichomonas foetus in transport, Diamond's and InPouch TF media.

The abilities of two isolates of Trtrichomonas foetus to survive and replicate in transport and Diamonds medium or in the InPouch TF system (Bio-Med Diagnostics) when exposed to different temperatures for different periods were determined in a series of experiments. Tubes containing thioglycollate transport medium or pouches were inoculated with 4000 to 5000 organisms and kept for up to seven days at 37°C, 22°C, 4°C, or -20°C. When the holding time had elapsed, the numbers of motile T foetus were counted. Samples in transport medium were transferred to Diamonds medium, and both the pouches and tubes containing Diamonds medium were incubated at 37°C. The cultures were examined and counted four or five times during the 10 to 14 day culture period. The sensitivity of the test under the different conditions, expressed as the number of positive cultures/the total number of samples × 100, varied from zero to 100 per cent depending upon the combination of variables considered. In each medium, with both isolates of T foetus, all samples kept for up to four days at 22°C or 37°C were positive. All cultures of samples kept more than five days at 4°C were negative. No positive cultures were detected when samples were kept more than three hours at -20°C. The day on which the cultures reached mean peak concentration varied with the temperature at which the samples had been kept before they were cultured.

Associations between oil- and gas-well sites, processing facilities, flaring, and beef cattle reproduction and calf mortality in western Canada.

From the fall of 1992 through calving 1996, detailed cow breeding outcome records were maintained actively for seven large cow-calf herds in western Canada. The numbers of mature females in the study for the breeding seasons beginning in 1992, 1993, 1994, and 1995 were 1122, 1177, 1251, and 1236, respectively. Outcomes included pregnancy status, calving interval, and the occurrence of twins, abortions, stillbirths, and neonatal mortality. Information also was collected on other risk factors known to influence beef-herd health and productivity. Detailed maps of active and inactive oil and natural-gas sites, batteries, compressor stations and processing plants were verified. Records of flaring activity at each facility were obtained from the government regulatory agency. Each flaring site then was classified as sour or sweet based on the presence or absence of hydrogen sulfide in the flared gas. A detailed inventory was prepared itemizing the type and number of facilities within 1.6 km (1 mile) of the center of each quarter section used for pasture. The total volume of gas flared within 1.6 km of the center of each pasture was determined for each month of the study. Appropriate risk periods where specified for each outcome and a cumulative exposure calculated for each breeding female (using detailed individual-animal records of cow movements between pastures and herd-management groups). Generalized estimating equations were used to evaluate the association between exposure and outcome and to adjust for potential confounders and clustering of binomial outcomes within herd. Increased risk of non-pregnancy was sometimes associated with exposure to one or more of the following facility types: sour-gas flaring battery facilities, all battery-flaring sites, active gas wells, and larger field facilities. The associations were not, however, consistent among years or even among risk periods for the same year. Facility proximity and flaring were not associated with increased abortion risk. Volume of flared sour gas from battery sites was associated with increased risk of stillbirth. Finally, sour-gas flaring was associated with increased calf-mortality risk for the 1992-1993 calf crop. Several examples of associations between exposure and increased productivity also were found (most of which involved either oil wells or all well sites).

A novel Ehrlichia genotype detected in naturally infected cattle in North America

During a research investigation to determine if cattle from British Columbia (BC), Canada were infected with Anaplasma marginale or other related rickettsial blood parasites, a novel Ehrlichia genotype was revealed. Blood from seven BC source cattle was bioassayed by intravenous inoculation into naïve splenectomised calves. Additional splenectomised calves were used as uninoculated negative control or A. marginale-inoculated positive control. Newly designed sets of primers specific for the msp5 gene of A. marginale or for the 16S rRNA gene were used to test blood samples collected from all source cattle and from all recipient calves prior to inoculation and up to 72 days post-inoculation. Results of the PCR assays as well as microscopic examination of stained blood smears failed to demonstrate A. marginale in any of the animals except for the positive control. The 16S rRNA PCR primers amplified DNA from samples from all BC source cattle, five of six of the corresponding recipient calves, and the A. marginale infected control animal. DNA sequence data indicated the presence of A. marginale only in the positive control calf. Blast analysis in GenBank showed that sequences of all other 16S rRNA PCR products clearly fit within the Ehrlichia genus in the Anaplasmataceae family which also includes members of the genus Anaplasma. Phylogenetic analyses using the 16S rRNA gene sequences strongly support the putative Ehrlichia organism as a distinct genotype with sequences of various strains of Ehrlichia canis as the closest clade. Ehrlichia ruminantium is the only other species of the genus known to naturally infect cattle, apart from the present Ehrlichia isolate. However, within the genus, E. ruminantium is phylogenetically the furthest removed species from the novel genotype. The finding of this novel Ehrlichia represents the first known natural ehrlichial infection in cattle in North America. Nevertheless, it is unclear whether cattle are an incidental or primary host, particularly since deer are recognized as reservoir hosts for other species of Ehrlichia. Although other Ehrlichia spp. are known to be pathogenic for animals and zoonotic, these features are presently unknown for this novel genotype.

Associations between total sulfation, hydrogen sulfide deposition, and beef-cattle breeding outcomes in western Canada

Beef cows sometimes are pastured close to oil- and gas-production and processing facilities. We measured the health and productivity of cow-calf herds with differing exposure to sour-natural-gas processing facilities in western Canada. In seven cow-calf herds, the numbers of eligible mature females included in the study for the breeding seasons beginning in 1993, 1994, and 1995 were 1177, 1251, and 1236, respectively. Outcomes included pregnancy status, calving interval, and the occurrence of twins, abortions, stillbirths, and neonatal mortality. Information also was collected on other risk factors known to influence beef-herd health and productivity. Monthly measurements from a network of passive air-monitoring devices were used to estimate exposure. Total sulfation and H2S deposition were used as markers for the complex mixture of compounds found in emissions from sour-gas processing plants and sour flares. Most herds were managed in multiple pasture groups. Cumulative exposure assessments were made from records of individual-animal movements between pastures. Generalized estimating equations were used to evaluate the association between exposure and outcome and to adjust for potential confounders and clustering of binomial outcomes within herd. No consistent associations were found between either total sulfation or H2S deposition and productivity parameters across the cow-calf production cycles. There were, however, five examples of significant associations between increasing cumulative exposure to total sulfation and decreased productivity in the 18 models examined.

Agreement between Three Serological Tests for Neospora Caninum in Beef Cattle

During 1999, serum samples were collected from beef cows on pastures in western Canada. Some of the herds had a history of confirmed abortions associated with Neospora caninum infection. All these samples were initially analyzed using a single application of 1 common commercial enzyme-linked immunosorbent assay (ELISA) for antibodies to N. caninum. From these initial results, 239 positive and 250 negative samples were randomly selected for further testing. This group of samples was retested using the 3 commercially available ELISA tests for N. caninum as per the manufacturers recommendations. The agreement between 2 of the ELISAs was good (k = 0.76); agreement of these 2 tests with the third test was much lower (k = 0.46 and 0.60). Quantitative agreement between tests measured by intraclass correlation coefficients was also acceptable between the first 2 tests but was almost zero when the first 2 tests were compared with the third. This information is necessary to understand the differences in seroprevalence reported in different regions from laboratories using different methods.

Surveillance for Cryptococcus gattii in horses of Vancouver Island, British Columbia, Canada

In the last decade Cryptococcus gattii has emerged as an important human and animal pathogen in southwestern British Columbia (BC), Canada. When the disease initially emerged it was identified in humans and multiple animal species on the east coast of Vancouver Island. From fall 2003 until summer 2004, active surveillance was initiated to look for horses exposed to or infected with the organism by performing nasal cultures and serum antigen testing in horses residing within 10 km of known areas of environmental reservoirs of the fungus. Surveillance efforts were facilitated by local equine practitioners who were also encouraged to report clinical cases. Nasal colonization was identified in four of the 260 horses tested but none had a serum cryptococcal antigen titer. All positive horses were from the same geographic area near Duncan, BC. During the study period, a single horse was diagnosed with systemic cryptococcosis and euthanized; clinical and post mortem information is described. As this organism continues to disseminate in the Pacific Northwest it is important for veterinarians to be familiar with the disease as early diagnosis may enable more effective treatment.

Characterising barrier function among regions of the gastrointestinal tract in Holstein steers

The objective of this study was to characterise the regional variation in the barrier function of the gastrointestinal tract in Holstein calves using the flux rates of mannitol and inulin as permeability markers and tissue conductance (Gt) as an electrophysiological indicator of barrier function. Six Holstein steer calves (6 months of age) fed a common diet were used. Calves were killed by captive bolt stunning and pithing, and tissues were collected from the rumen, omasum, duodenum, jejunum, ileum, caecum, proximal colon, and distal colon. Tissues were carefully washed using a pre-heated (38.5°C) buffer solution (pH 7.4) saturated with oxygen and then transported to the laboratory. The mucosa was prepared by hand stripping and mounted between two halves of an Ussing chamber (n = 3/region with an exposed surface area of 3.14 cm2 for rumen and omasum and 1 cm2 for all other tissues). All tissues were incubated under short-circuit conditions and exposed to a similar buffer solution except for the energy source; rumen, omasum, caecum, and colon tissues were incubated with buffer containing short-chain fatty acids while tissues from the small intestine were bathed in buffer containing glucose. The Gt and the serosal-to-mucosal flux rates of 14C-inulin and 3H-mannitol were measured as indicators of barrier function. The serosal-to-mucosal flux rate of mannitol was greatest (P < 0.001) in the jejunum [104.8 nmol/(cm2 × h)] and least in the rumen and omasum [20.3 and 18.6 nmol/(cm2 × h), respectively]. In contrast, the serosal-to-mucosal flux rate of inulin was greatest (P < 0.001) in the omasum [158.6 nmol/(cm2 × h)] followed by the rumen [87.3 nmol/(cm2 × h)] with no differences among the other regions [18.7 – 62.0 nmol/(cm2 × h)]. The Gt was greatest (P < 0.001) in the jejunum (34.6 mS/cm2) and least for the rumen (3.67 mS/cm2) and omasum (3.23 mS/cm2). The Gt was correlated with both inulin and mannitol flux rates in duodenum, caecum and proximal colon (P < 0.05); whereas, no such correlations existed in jejunum, ileum and distal colon. The Gt was correlated with the mannitol flux rate but not the inulin flux rate in rumen and omasum. For all regions but the rumen and omasum there was a positive correlation between mannitol and inulin flux rates. These data indicate that the translocation of a large molecule (inulin) across the omasum and rumen is greatest despite having an apparently tight epithelium based on Gt and mannitol flux rate, while the jejunum appears to have greatest potential for paracellular permeability.