Flow cytometric analysis of intracellular complexity and CD45 expression for use in rapid differentiation of leukocytes in bovine blood samples.

OBJECTIVE: To develop an efficient and reliable method that accurately differentiates bovine lymphocytes from monocytes in leukograms. SAMPLE POPULATION: Blood samples from 30 healthy cows and 1 calf with bovine leukocyte adhesion deficiency. PROCEDURE: Flow cytometric analysis of intracellular complexity and CD45 expression on bovine leukocytes was compared with results for conventional light microscopy methods. Verification of leukocyte subpopulations determined by intracellular complexity and CD45 expression was conducted, using 2-color phenotypic analysis with selected monoclonal antibodies. RESULTS: The CD45 and side-scatter properties of bovine leukocytes clearly differentiated cell types, including neutrophils, eosinophils, monocytes, and lymphocytes. CONCLUSIONS AND CLINICAL RELEVANCE: This is a rapid assay that is simple to use. More importantly, it is more accurate than the conventional method that involves the use of blood slides and light microscopy, because of the ability of the assay to readily distinguish bovine monocytes and lymphocytes. Rapid preparation of samples and short analysis times allow for efficient and reliable examination of a large number of samples, and the task of viewing slides by light microscopy is eliminated. The labor-savings benefit of this procedure is most apparent in research environments that require frequent processing of batches of blood samples.     

Long-term variation in brown trout, Salmo trutta L., stocking success in a large lake: interplay between availability of suitable prey and size at release

Abstract –  Factors affecting long-term variation in brown trout, Salmo trutta L., stocking success were examined in a large lake, Lake Oulujärvi, in central Finland. Brown trout were stocked in spring (late May to early June) in 1974–1991 and in summer (late June to early July) in 1992–2001. The biomass of the vendace, Coregonus albula (L.), population (prey) at release time had the largest positive effect on stocking success within both periods: biomass of adult vendace in spring and both 0+ and adult vendace in summer. Increasing the size of stocked fish had a positive effect if the vendace available at release were only adults. The increasing trend of predator-catch-per-unit-effort (CPUE) [combined CPUE of northern pike Esox lucius L., burbot Lota lota (L.), and pike-perch Stizostedion lucioperca (L.)] through the study period and its negative effect on trout stocking success suggested an increasing effect of predation within the entire time series.     

Periodontal disease. Therapy and prevention

Success in veterinary dentistry lies in good home care and regular, thorough dental prophylaxis. The removal of supragingival and subgingival calculus must be accomplished before periodontal disease can be controlled. When gingival sulcus depths exceed 4 or 5 mm, the gingivae must be excised or reflected to allow proper treatment of the pocket area. Antibiotic therapy should be instituted in the extensively involved veterinary dental patient.     

Vaccination of the brushtail possum (Trichosurus vulpecula) against Mycobacterium bovis infection with bacille Calmette-Guérin: the response to multiple doses.

In New Zealand, the brushtail possum (Trichosurus vulpecula) is the principal wildlife vector of bovine tuberculosis. Control of infected possum populations contributes to the control of tuberculosis in domestic livestock. Vaccination is potentially a complementary strategy to population control, but to be cost-effective, administration of the vaccine to possums would need to be from an appropriately designed automatic vaccinator. Possums themselves would activate the vaccinator so that it would deliver an aerosol spray of vaccine. There would be no direct way to prevent possums receiving multiple doses of vaccine. This study examined the effect on protective immunity of repeated vaccination. Captive possums were vaccinated with BCG strain pasteur 1173P2 either 12 times at weekly intervals, twice at 6-weekly intervals, or once. Vaccination was by a combination of intranasal aerosol and conjunctival instillation. Eight weeks after the last dose of vaccine, all possums were challenged intratracheally with Mycobacterium bovis strain 83/6235. Vaccination induced a significant immune response as measured by the lymphocyte proliferation assay (LPA). A significant level of protection, as measured by the response to challenge, developed in all the vaccinated possum groups, but protection was greatest in the group vaccinated 12 times. It was concluded that protection would be enhanced if vaccinations were repeated at short intervals (weekly), but no benefit or detriment resulted from revaccination after longer intervals (1-2 months).     

Intramuscular and oral disposition of enrofloxacin in African grey parrots following single and multiple doses

The intramuscular (IM) and oral (PO) disposition of enrofloxacin, a new fluoroquinolone antimicrobial drug, were evaluated in African grey parrots. Peak enrofloxacin concentration, mean (+/- SEM), at 1 h following a 15-mg/kg IM dose was 3.87 (+/- 0.27) micrograms/ml and declined with a mean residence time of 3.05 h. Peak enrofloxacin plasma concentrations at 2 to 4 h following oral doses of 3, 15, and 30 mg/kg were 0.31 (+/- 0.11), 1.12 (+/- 0.11), and 1.69 (+/- 0.23) micrograms/ml, respectively, and declined with a mean residence time of 3.44-5.28 h. The relative bioavailability of the 15-mg/kg oral dose was 48%. An equipotent metabolite, ciprofloxacin, was detected in plasma at concentrations ranging from 3 to 78% of those of enrofloxacin. Enrofloxacin concentrations and area under the curve were significantly lower, the mean residence time significantly shorter and the ciprofloxacin/enrofloxacin ratios higher, following 10 days of oral treatment at 30 mg/kg every 12 h. Following 10 days of treatment, no significant biochemical changes were noted; however, polydipsia and polyuria occurred in treated birds, but resolved quickly upon discontinuation of enrofloxacin administration. These studies indicate that a rational starting dose for enrofloxacin in psittacines (7.5-30 mg/kg BID) should be higher than those in other domestic animals.     

Effect of heterogeneous digesta chemical composition on the accuracy of measurements of fiber flow in dairy cows.

The objective of the current study was to evaluate the effect of unrepresentative sampling of digesta particulate matter entering the omasal canal on the accuracy of fiber flow measurements. The experimental design comprised one period, one diet, and three cows as experimental units. Within each cow, the physical and chemical composition of digesta particulate matter was assessed at seven sites within the digestive tract. Three Finnish Ayshire dairy cows, equipped with ruminal and simple-T duodenal cannulas, in extended lactation were offered grass silage twice daily on an ad libitum basis. Digesta samples were collected from the rumen (dorsal and ventral sac), reticulum, omasal canal, omasum, duodenum, and rectum to determine particle size distribution in digesta, chemical composition of various particle size fractions, and distribution of two flow markers (Cr-labeled straw and indigestible NDF [INDF]) among particle size fractions. Digesta samples were wet-sieved using sieves of 2.50, 1.25, 0.630, 0.315, 0.160, and 0.080 mm. Particulate matter was analyzed for OM, NDF, and Cr concentrations, and INDF concentration was determined based on 12-d ruminal incubation. The particle size of digesta entering the omasal canal was larger compared with the omasum or the duodenum, suggesting that omasal canal samples were not representative of particle size distribution truly escaping the rumen. The concentration of potentially digestible NDF (PDNDF) decreased with decreasing particle size. The PDNDF concentration of particulate matter retained on all sieves was greatest in the rumen and gradually decreased along the digestive tract. From the reticulorumen to the omasum, the decrease was associated with decreased particle size, reflecting selective passage of particulate matter. In contrast, from the omasum to the duodenum and rectum, the PDNDF concentration decreased within each particle size fraction without effect on particle size, indicating a nonselective passage of particulate matter between these sites. Variation between particle size fractions was slightly greater for Cr concentration than for INDF concentration, indicating that unrepresentative sampling of particulate matter had a greater effect on Cr concentration compared with that of INDF. Owing to unrepresentative sampling, NDF entering the omasal canal was overestimated by 5% using INDF and underestimated by 7% using Cr as a particle phase marker. Of total NDF digestibility, proportionally 0.90, 0.07, and 0.03 occurred in the reticulorumen, omasum, and intestines, respectively. The current results indicate that, despite unrepresentative sampling of digesta particulate matter entering the omasal canal, the errors in determined NDF flow were small. The omasum may have a greater role in postruminal NDF digestion than the intestines.     

A field study on reproductive efficiency of mares maintained predominately on native pasture

A field study utilizing 145 mares of various age and breeding status was conducted to determine reproductive performance under native range conditions with only limited supplemental feeding. All mares had an average initial body condition score of 4.5 and a final score of 5.1 by the time breeding was completed and mares returned to pasture. Average foaling rate was 80%, and mares that had not conceived during the previous breeding season had a foaling rate of 94%, which was higher (P<.05) than 74% for lactating mares. Mares 16 years and older had a significantly lower foaling rate (P<.05) than younger mares. Old mares that were lactating at time of breeding had only a 37% foaling rate, which was less (P<.05) than for young lactating mares. The 94 mares bred by natural cover or artificial insemination that actually foaled required 1.43 cycles per conception. Lactating mares in the oldest age group required more cycles per conception (P<.05) than open mares, and these older, lactating mares also required more cycles per conception than younger mares with foals at side. Those mares diagnosed as pregnant or open at 45 days post breeding had a pregnancy rate of 97%. Average pregnancy loss for all mares was 7.7%. These data indicate that lactating mares in moderate body condition tended to skip a breeding season and that a body condition score of 5 was only marginally acceptable, especially in the case of lactating mares. Authors' address: Equine Science Program, Department of Animal Science, Texas Agricultural Extension Service, Texas A&M University, College Station, TX 77843. Technical Article Number 30023, Texas Agr. Exp. Sta.