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.     

Genetic diversity of herbicide-resistant and -susceptible Avena fatua populations in North Dakota and Minnesota

Genetic diversity within and among 20 herbicide-resistant (HR) and 16 herbicide-susceptible (HS) Avena fatua multi-field populations was determined using 82 polymorphic loci resulting from two intersimple sequence repeat (ISSR) primers and one long-primer random amplified polymorphic DNA (LP-RAPD) primer. Collections from the Red River Valley of North Dakota and Minnesota, sampled in 1964 and 2000, represented A. fatua populations before and after intensive exposure to herbicides. A 1995 collection from south-west North Dakota represented A. fatua exposed to low herbicide selection. Despite differences in years of herbicide exposure among collections, both HR and HS populations from every collection maintained nearly similar levels of ISSR and RAPD diversity. Genetic differentiation among populations (G_ST) varied from 11% to 13% among HR populations and from 9% to 16% among HS populations, indicating that 84–91% of total variation remained within HS or within HR populations. Minimal difference in gene diversity between HR and HS is consistent with multiple origins of resistance, where HR A. fatua most likely evolved from diverse founding individuals.     

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.     

Histopathological and hematological findings in myeloid leukemia induced by a new feline leukemia virus isolate

Myeloid leukemia was induced by a new feline leukemia virus isolate FeLV-AB/GM-1 in a high proportion of cats. The latency period was short. Three to 5 weeks after infection early changes were detectable in the bone marrow, and cats developed leukemia 5 to 8 weeks after infection. The results of the present histological and cytological studies suggested that there were two stages in the development of leukemia. The first stage appeared to be equivalent to the syndrome of bone marrow dysplasia or preleukemia which, however, converted rapidly to leukemia. Cytopenia(s) were the main hematological findings in all preleukemic and leukemic cats. White blood cell counts were low or normal, but the number of leukemic and abnormal cells increased in the peripheral blood with the progression of the disease. This reliable model system lends itself to further studies to elucidate the pathogenesis of myeloproliferative disorders.     

Survival of Leptosphaeria maculans in soil on residues of Brassica napus in South Australia

The survival of Leptosphaeria maculans , which causes phoma stem canker (blackleg), on oilseed rape residues ( Brassica napus ) in South Australia was investigated. Using a quantitative polymerase chain reaction (PCR) assay for L. maculans DNA, the pathogen was mainly detected in the upper 5 cm of the soil profile, including residues on the soil surface. As the size of organic matter particles in the soil decreased, so did the quantity of L. maculans detected in them. To obtain representative data for a field, at least 30 subsamples needed to be collected over the 0·81 ha area studied. In a survey of 49 commercial fields in South Australia, most L. maculans was detected in fields 1 year after oilseed rape had been grown, with less detected after 2 years and negligible amounts 3 years or more after cropping. The diagnostic DNA-based assay for L. maculans reduced the time and cost of studying L. maculans survival in soil and increased the sensitivity and accuracy of results compared with estimates of propagule number of colony-forming units on a semiselective medium.     

Effects of pesticides and fertilizer on invertebrate populations of grass and wheat plots in Kent in relation to productivity and yield

At a site in Kent, perennial ryegrass (Lolium perenne) variety S24 and Italian ryegrass (L. multiflorum) variety RVP, wheat varieties Armada and Norman, and the original mixed grass ley were grown in small plots during 1982–84. Two toxic pesticides (phorate and aldicarb) were applied to half the total number of plots three times each year to eliminate soil invertebrate populations. Fertilizer was also applied to most plots. The yields of the crops, grown with and without pesticide, and the effects of the fertilizer were compared. Grass herbage yield was measured on three occasions during the summers of 1983 and 1984. Wheat grain yields were also determined in 1983. During the first year significant differences were not apparent in grass dry matter yield between pesticide-treated and non-treated plots, but significant differences were found in the second year. The perennial ryegrass was more susceptible to pest damage than the Italian ryegrass or the grass ley. Grass yields varied between cuts and in relation to variety and pesticide treatment, yields tending to be greater in untreated plots. Fertilizer treatment greatly increased grass dry matter yields, particularly with the Italian ryegrass. The effects of pesticide treatment on both wheat varieties varied although some yield enhancement was evident. Invertebrate animal populations in pesticide and fertilizer-treated plots were also assessed in autumn 1982, spring and autumn 1983 and spring 1984. In contrast to pesticide treatment, fertilizer treatment had little effect on soil invertebrate populations. Nematode populations were reduced at each sampling occasion by the pesticide treatment. Slug populations were initially unaffected but were subsequently reduced. Leatherjackets, by far the most abundant pest in both grass and wheat plots, were markedly affected by pesticides on all sampling occasions. Generally, fewer soil-dwelling dipterous larvae were recovered in spring than in autumn. Stem-boring dipterous larvae were virtually absent.