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.     

Ultrasonographic and Gross Pathological Findings in the Mammary Glands of Weaned Sows Having Suffered Recidiving Mastitis Metritis Agalactia

In a large pig breeding herd with high prevalence of post-parturient diseases of the sows, weaned sows of different parity groups with (n = 663) or without (n = 1125) recidiving mastitis metritis agalactia (MMA) in their previous history were subjected to ultrasonography. A total of 114 of 663 sows with recidiving MMA in their previous history and with ultrasonographic visible mammary gland changes, and 157 of sows without recidiving MMA in their previous history were culled and subjected to gross pathological and bacteriological examination of their mammary glands. The sows having suffered MMA had more (p < 0.001) hyperechogenic images in their mammary glands compared with the sow having suffered no recidiving MMA. Abdominal glands were more (p < 0.01) prone to pathological changes compared with the pectoral ones. Sows of high parity had more hyperechogenic images and gross pathological changes in their mammary glands compared with the sows of low parity.     

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).     

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.     

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.     

Bovine spongiform encephalopathy in Northern Ireland: epidemiological observations 1988-1990.

This study describes the epidemiological features of bovine spongiform encephalopathy (BSE) in Northern Ireland where the first case occurred in November 1988. They were very similar to those observed in Great Britain except that the annual incidence of BSE in 1990 in Northern Ireland, 2.3 confirmed cases per 10,000 adult cows, was approximately one 10th of that in Great Britain. The findings were also consistent with the current hypothesis that affected cattle had been exposed to a scrapie-like agent via cattle feedstuffs containing ruminant-derived protein. However, a preliminary investigation of the potential sources of infection for cattle in Northern Ireland did not provide any conclusive evidence.     

ULTRASONOGRAPHY OF ALIMENTARY LYMPHOSARCOMA IN THE CAT

Clinical, radiological and ultrasonographic findings from 22 cats with gastrointestinal lymphosarcoma were reviewed. The most common clinical findings were anorexia, weight loss, vomiting, lethargy, depression, fever, anemia and a palpable abdominal mass. Abdominal radiographs of 12 cats revealed a mass associated with the gastrointestinal tract and/or mesentery, and decreased serosal detail, suggestive of peritoneal effusion. The most common ultrasonographic finding was transmural gastric or intestinal thickening associated with loss of normal wall layering, reduced wall echogenicity and localized hypomotility. As is noted in people, other patterns such as transmural-segmental, transmural-nodular, transmural-bulky and mucosal infiltration were represented. In 3 cats, mesenteric lymph-adenopathy was responsible for most of the mass-effect. Fine-needle aspiration biopsy, automated microcore biopsy, endoscopic biopsy or necropsy confirmed the diagnosis of lymphosarcoma in all cats. Chemotherapy was instituted in 19 cats. Only 6 cats had a complete response to treatment. The results of this study indicate that ultrasonography is a valuable tool for the diagnosis of feline alimentary lymphosarcoma and that biopsies can be obtained under ultrasonographic guidance.