Individual antigens of cattle. Bovine CD2 (BoCD2).

The data obtained in the workshop provide further evidence that CH128A and IL-A26 and the 12 new mAbs that form a cluster recognise the bovine orthologue of CD2. The mAbs inhibit rosetting with SRBC, stain cells in primary and secondary lymphoid organs in patterns consistent with those obtained in humans with anti-CD2 mAbs, and the 11 IgG mAbs all immunoprecipitate a peptide with a Mr of 58-62 kDa. It is not clear from the studies whether the epitopes defined by the mAbs correspond with the region I and II epitopes present on CD2. None of the data suggest that any of the mAbs recognise the region III (CDD2R) epitope (Peterson and Seed, 1987; Knapp et al., 1989). Further studies are now needed to define the physical and functional relation of the epitopes and establish whether antibody-mediated activation corresponds with that noted in humans. Data reported in one study (Baldwin et al., 1988) with IL-A26 suggest possible differences in the requirements for activation. In addition, further studies are needed to demonstrate how many cell types express BoCD2. In mice, evidence has been presented which shows the mouse orthologue is expressed on some B cells (Yagitta et al., 1989). Studies in cattle have clearly shown CD2 is present on the majority of CD4+ and CD8+ T-cells and a small population of CD4-/CD8- cells (Baldwin et al., 1988; Davis, unpublished observations). Evidence presented in this workshop has shown that some CD2+ cells express a WC2 molecule (Sopp et al., 1991).(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Culture of bovine bone marrow progenitor cells in vitro.

In vitro methylcellulose cultures of bovine bone marrow progenitor cells were developed. An existing technique described for bovine species was compared to a method for human tissue and further adapted during subsequent experiments. Bovine bone marrow samples were collected at the slaughterhouse, and mononuclear cells were separated by gradient centrifugation (1.077 g/ml specific density and 400 g). The use of 3% bovine leucocyte-conditioned medium, produced by stimulation of blood lymphocytes with 4 microg/ml concanavalin A and harvested on day 4 of culture, gave better results than the use of supernatant of the human bladder carcinoma 5637, which is widely used in human bone marrow cultures. However, bovine leucocyte-conditioned medium was not added to erythroid cultures because inhibitory effects were observed. Erythroid colonies were stimulated with erythropoietin, and hemin was added to enable microscopic identification. Reduced oxygen tension was necessary to induce growth of erythroid colonies. This was not necessary for myeloid cultures. In conclusion, the results of this study show that the growth of myeloid and erythroid colonies in methylcellulose-based medium requires different culture conditions, which are different from the culture conditions for human cells.     

Leeching as initial treatment in a cat with polycythaemia vera.

Polycythaemia vera was diagnosed in a three-year-old domestic shorthaired cat referred because of seizures and a high packed cell volume (PCV). Laboratory examination revealed severe erythrocytosis (PCV 79 per cent). Diagnosis was reached by excluding causes for relative and secondary absolute polycythaemia. As phlebotomy proved impossible for initial treatment due to hyperviscosity, four leeches were used to suck blood and the PCV was consequently reduced to 64 per cent. A further 24 hours later, when bleeding at the sites of sucking had stopped, the PCV was 56 per cent. Long-term management of the condition was achieved with hydroxyurea (100 mg/cat once daily) and intermittent phlebotomy. Initial treatment using leeches in cases of polycythaemia vera is a simple, non-invasive, well tolerated and effective method where phlebotomy is not possible.     

Pharmacokinetics and pharmacodynamics of butorphanol in llamas after intravenous and intramuscular administration.

OBJECTIVE: To evaluate disposition of butorphanol after i.v. and i.m. administration, effects on physiologic variables, and analgesic efficacy after i.m. administration in llamas. DESIGN: Nonrandomized crossover study. ANIMALS: 6 healthy adult male llamas. PROCEDURE: Butorphanol (0.1 mg/kg [0.045 mg/lb] of body weight) was administered i.m. first and i.v. 1 month later. Blood samples were collected intermittently for 24 hours after administration. Plasma butorphanol versus time curves were subjected to pharmacokinetic analysis. Two months later, butorphanol (0.1 mg/kg) was administered i.m., and physiologic variables and analgesia were assessed. RESULTS: Extrapolated peak plasma concentrations after i.v. and i.m. administration were 94.8 +/- 53.1 and 34.3 +/- 11.6 ng/ml, respectively. Volume of distribution at steady state after i.v. administration was 0.822 +/- 0.329 L/kg per minute and systemic clearance was 0.050 +/- 0.014 L/kg per minute. Slope of the elimination phase was significantly different, and elimination half-life was significantly shorter after i.v. (15.9 +/- 9.1 minutes) versus i.m. (66.8 +/- 13.5 minutes) administration. Bioavailability was 110 +/- 49% after i.m. administration. Heart rate decreased and rectal temperature increased. Somatic analgesia was increased for various periods. Two llamas became transiently sedated, and 2 became transiently excited after butorphanol administration. CONCLUSIONS AND CLINICAL RELEVANCE: Although i.v. administration of butorphanol results in a short half-life that may limit its analgesic usefulness, the elimination half-life of butorphanol administered i.m. is likely to be clinically useful. The relationship among plasma butorphanol concentration, time, and analgesia differed with the somatic analgesia model; clinically useful analgesia may occur at lower plasma concentrations than those reported here.     

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.     

The Effect of Exogenous Hormone Treatment on Spermiation in Rhynchocypris oxycephalus (Sauvage and Dabry)

For the evaluation of hormonal control of spermiation in fish, a method to quanify the spermiation response of mature Rhynchocypris oxycephalus (Sauvage and Dabry) to hormonal therapy is described. Spermatocrit was determined after 7 min centrifugation at 18,000 ± g and sperm density was estimated by a standard hemocytomer method. Sperm density can be predicted from spermatocrit since their relationship is linear as described by the regression equation, Y = 3.68X - 27.18 ( R ² = 0.82, N = 50). where Y is spermatocrit and × is sperm density. Milt production by mature R. oxycephalus was highest at 24 h after injection of 1,000 IU human chorionic gonadotropin (HCG) and 50 μg luteinizing hormone-releasing hormone analogue (LHRHa) per kg body weight. Increased milt production coincided with low spermatocrit and sperm density levels. These results demonstrate that spermiation in mature R. oxycephalus can be reliably evaluated by a spermatocrit method and that HCG and LHRHa are effective in stimulating of spermiation in this species.     

The Application of GIS in Small Watershed Classification in Loess Plateau

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