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.     

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.     

Fate of Escherichia coli and Escherichia coli O157 in soils and drainage water following cattle slurry application at 3 sites in southern Scotland

Abstract. Slurry from farm animals may contaminate water supplies, rivers and bathing waters with faecal coliforms, such as Escherichia coli . Where animals harbour the O157 strain the hazard to human health is particularly high, but both the hazard level, and the low incidence and sporadic nature of the excretion of E. coli O157 make it difficult to study this strain under field conditions. The survival of total E. coli and of E. coli O157 were compared in the laboratory for two soils under controlled temperature and moisture. E. coli O157 die-off rate was the same as or quicker than for total E. coli . This result meant that field experiments studying the fate of total E. coli should give a satisfactory evaluation of the risk of water contamination by the O157 strain. In four field experiments at three sites, slurry containing total E. coli numbers of 2.2 × 10⁴ to 5.7 × 10⁵ colony forming units per mL (c.f.u. mL^–1) was applied to drained field plots. Field die-off was faster than expected from laboratory experiments, especially in one experiment where two weeks dry weather followed application. In all but this experiment, the first drain flow events after slurry application led to very high E. coli concentrations in the drains (10³ to 10⁴ c.f.u. mL^–1). E. coli O157 was present in the slurry used for two of the experiments (33 c.f.u. per 100 mL in each case). However the proportion of E.coli O157 was very low (about 1 in 10⁵) and it was not detected in the drainage water. After the first week E. coli drainage water numbers decreased rapidly but they were 1–10 c.f.u. mL^–1 for much of the sampling period after slurry application (1–3 months).     

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.     

0–7–21 Radiation Therapy for the Treatment of Canine Oral Melanoma

Eighteen dogs with malignant melanoma of the oral cavity were treated with high-dose per fraction (0–7–21) radiation therapy. Eight hundred cGy was administered on days 0, 7, and 21 for a total dose of 2,400 cGy in 3 weeks. Of 17 dogs evaluated, 9 (53%) had a complete remission and 5 (30%) achieved a partial remission with an overall response rate of 83%. Local failure occurred in 2 of the 9 dogs where a complete response was initially observed. One dog died of intercurrent disease, and one died of metastatic disease without evidence of local recurrence. Five dogs are alive and free of disease 9 to nineteen months from the initiation of therapy. The 0–7–21 protocol was well-tolerated, and acute radiation reactions were low-grade and limited to the skin. The results of this study demonstrate that oral melanomas in dogs are responsive to radiation. 0–7–21 radiation therapy offers a viable alternative to radical excision, especially when tumor volume or location would require cosmetically or functionally debilitating surgery.     

Effect of Split Nitrogen Application on Growth and Yield of Wheat (T. aestivum L.) Genotypes with Different N-Assimilation Potential

A field experiment was conducted with different nitrogen regimes to assess the growth and yield performance of wheat genotypes which differ in nitrate assimilation potential. Genotypic differences in biomass accumulation were observed at different growth stages. The nitrogen treatment had little effect on biomass accumulation at early stages of growth, while at later stages of growth there was enhanced biomass accumulation when N was applied in more than two splits. On an average, genotypes with high nitrate reductase activity (the 'HNR' genotypes) accumulated 14.2 % more biomass than the genotypes with low nitrate reductase activity ('LNR' genotypes) when an extra dose (40 kg N ha⁻¹) of nitrogen was given at the time of anthesis. The application of nitrogen in more than two splits increased grain yield of both 'HNR' and 'LNR' genotypes mainly by increasing grain weight per ear. The application of an extra dose of nitrogen (40 kg N ha⁻¹) at the time of anthesis increased grain yield of 'HNR' genotypes by 38.5 % as compared to 'LNR' genotypes.     

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.