Evaluation of enzyme-linked immunosorbent assays performed on milk and serum samples for detection of paratuberculosis in lactating dairy cows

OBJECTIVE: To determine whether results obtained for milk and serum samples with ELISAs intended for diagnosis of paratuberculosis in dairy cows were comparable to results obtained by means of mycobacterial culture of fecal samples. DESIGN: Cross-sectional study. ANIMALS: 689 lactating dairy cows in 9 Ontario herds. PROCEDURE: Milk, serum, and fecal samples were obtained from all cows. Fecal samples were submitted for mycobacterial culture. Serum samples were tested with a commercially available ELISA for antibodies against Mycobacterium paratuberculosis, and preserved milk samples were tested with an indirect ELISA for antibodies against M paratuberculosis. RESULTS: Results were positive for 130 of the 689 (18.9%) serum samples, 77 of the 689 (11.1%) milk samples, and 72 of the 689 (10.4%) fecal samples. The level of agreement between results for milk and serum samples was only moderate. Proportions of positive results for serum and fecal samples were significantly different, but proportions of positive results for milk and fecal samples were not significantly different. In addition, results for milk samples had a higher level of agreement with results of mycobacterial culture than did results for serum samples. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the indirect ELISA used on milk samples may be a convenient method of detecting paratuberculosis in dairy herds.     

Parasitemia in an immunocompetent horse experimentally challenged with Sarcocystis neurona sporocysts

Equine protozoal myeloencephalitis (EPM) is a serious neurological disease of horses in Americans. Most cases are attributed to infection of the central nervous system with Sarcocystis neurona. Parasitemia has not been demonstrated in immunocompetent horses, but has been documented in one immunocompromised foal. The objective of this study was to isolate viable S. neurona from the blood of immunocompetent horses. Horses used in this study received orally administered S. neurona sporocysts (strain SN 37-R) daily for 112 days at the following doses: 100/day for 28 days, followed by 500/day for 28 days, followed by 1000/day for 56 days. On day 98 of the study, six yearling colts were selected for attempted culture of S. neurona from blood, two testing positive, two testing suspect and two testing negative for antibodies against S. neurona on day 84 of the study. Two 10 ml tubes with EDTA were filled from each horse by jugular venipuncture and the plasma fraction rich in mononuclear cells was pipetted onto confluent equine dermal cell cultures. The cultures were monitored weekly for parasite growth for 12 weeks. Merozoites grown from cultures were harvested and tested using S. neurona-specific PCR with RFLP to confirm species identity. PCR products were sequenced and compared to known strains of S. neurona. After 38 days of in vitro incubation, one cell culture from a horse testing positive for antibodies against S. neurona was positive for parasite growth while the five remaining cultures remained negative for parasite growth for all 12 weeks. The Sarcocystis isolate recovered from cell culture was confirmed to be S. neurona by PCR with RFLP. Gene sequence analysis revealed that the isolate was identical to the challenge strain SN-37R and differed from two known strains UCD1 and MIH1. To our knowledge this is the first report of parasitemia with S. neurona in an immunocompetent horse.     

Concentration of methylprednisolone in the centrodistal joint after administration of methylprednisolone acetate in the tarsometatarsal joint

REASONS FOR PERFORMING STUDY: The centrodistal (CD) and tarsometatarsal (TMT) joints are often injected individually with a corticosteroid to resolve lameness caused by osteoarthritis (OA). There are no data available regarding diffusion of methylprednisolone (MP) from the TMT joint to the CD joint. HYPOTHESIS: A therapeutic concentration of MP diffuses into the CD joint after methylprednisolone acetate (MPA) is administered into the TMT joint. OBJECTIVE: To measure the concentration of MP in the CD joint after MPA was administered into the TMT joint. METHODS: MPA was administered into a TMT joint of 16 horses. At different times, the ipsilateral CD joint of these horses was injected with a small amount of saline and recovered saline was measured for concentration of MP using high performance liquid chromatography. RESULTS: Six hours after administration of MPA into the TMT joint, a therapeutic concentration of MP was found in all 10 CD joints sampled at this time. CONCLUSIONS: Horses with pain arising from the distal 2 joints of the hock can be treated by administering MPA into the TMT joint alone. POTENTIAL RELEVANCE: Administering MPA into the TMT joint only, to treat OA of the distal 2 hock joints, reduces the difficulties and risks associated with centesis of the CD joint.     

Synthetic myosin filaments

A stable preparation of myosin filaments was formed in a medium at pH 8.0. The filament length varied from 0.2 to 0.5 micron. Most of the material sedimented at 21S, but there was a minor peak (due to monomer) at 6.8S. The filaments did not taper and had large bulbous irregularities at the ends.     

Natural control of the cereal cyst nematode, Heterodera avenae Woll., by soil fungi at three sites

Populations (eggs/g dried soil) of the cereal cyst nematode Heterodera avenae Woll. failed to increase at two sites on susceptible spring barley but increased slightly at another on spring oats. Nematode multiplication was not related to the numbers of females produced on roots in summer; fewer female nematodes were produced where populations increased than where they declined. The failure of the nematode to multiply was due to few (less than 40%) of the females forming cysts containing eggs, a reduction in fecundity and an increase in the number of encysted eggs containing fungi. Verticillium chlamydosporium was the main fungus causing the decline in nematode numbers. Nematophthora gynophila was present at all sites but was most active at Woburn where irrigation was used to keep soils moist. V. chlamydosporium was the main parasite of encysted eggs at all sites but soil-borne saprophytic fungi were also isolated. These fungi were not found in many eggs and were considered to be only weak parasites. Differences in resting fungal spore densities in soil were not always related to differences in rates of parasitism. The activity of N. gynophila is affected by soil moisture and density of females as well as the density of spores in soil. The production of chlamydospores by V. chlamydosporium may also depend on the isolate of fungus present: some isolates produce no such spores on artificial media or, possibly, in soil. This work has shown that V. chlamydosporium and N. gynophila can reduce cereal cyst nematode populations under susceptible cereals but it is difficult to predict at present if they will give an effective control in a given soil or season.     

Duration of wing flapping in chickens shackled before slaughter

Shackling can induce wing flapping in chickens. In this survey of 30 chicken processing plants the proportion of birds flapping their wings had usually subsided to less than 0.5 per cent after the birds had been conveyed 2 m away from the shackling area. Thereafter, there were many causes of wing flapping, of which the following were thought to be important; tight fitting shackles on the birds' shanks, electric shocks at the water bath stunner before stunning, sudden bright sunlight, bends in the line, temporary loss of visual contact between neighbouring birds, and unevenness in the line which jolted the birds. If it is decided that legislation is required to prescribe a minimum 'hang-on' period before stunning it is suggested that birds should not be put through the stunner less than 12 seconds after shackling because after this time 99.7 per cent of the birds had stopped flapping.     

The vitamin A and vitamin E status of horses raised in Alberta and Saskatchewan.

The purpose of the study was to determine normal baseline levels of vitamin A and vitamin E in clinically normal horses under typical field conditions in Saskatchewan and Alberta. Heparinized blood samples were collected from approximately 400 clinically healthy horses selected from 24 locations in Alberta and Saskatchewan during a two-year period. For each horse, historical information including feed type, vitamin supplementation, time of year, sex, and age were recorded. From each blood sample, the plasma vitamin A (all-transretinol) and vitamin E (alpha-tocopherol) levels were measured using high pressure liquid chromatography. Normal baseline plasma vitamin A and vitamin E concentrations recorded during the study were 0.70 mumol/L and 7.65 mumol/L, respectively. The plasma vitamin concentrations were lower in the younger horses. The plasma vitamin levels were higher from May to August, as compared to other times of the year. Horses grazing fresh pasture exclusively during the summer months had plasma vitamin A and vitamin E concentrations that were 27% and 63% greater than horses fed harvested or stored feeds during the same time period. Sex-related differences were not evident in the study. A number of factors may influence the baseline plasma vitamin A and vitamin E levels in horses. Consequently, it is unadvisable to use a single evaluation to assess vitamin status. Multiple sampling from individual horses or sampling from many horses within a herd may reduce the variability and improve the ability to monitor vitamin status from plasma submissions.     

Repetitive propofol administration in dogs and cats

A bolus of propofol was administered to 10 dogs (6 mg/kg intravenously [IV]) and 10 cats (10 mg/kg IV) on three consecutive days. The occurrence of apnea, heart and respiratory rates, blood pressure, time to movement, and changes in a complete blood count and biochemical profile were recorded. Apnea was not seen in the dogs but was seen in three cats. Slight increases in the number of Heinz bodies were seen in six cats, but the increases were not considered clinically significant. No apparent cumulative adverse effects were seen from a bolus of bisulfite-containing propofol, administered on three consecutive days.