Differentiation of Staphylococcus sciuri Strains Isolated from Free‐Living Rodents and Insectivores

Twenty‐nine Staphylococcus sciuri strains isolated from free‐living insectivores and rodents were comparatively analysed for their biochemical capacities and their SmaI macrorestriction patterns. The 29 S. sciuri isolates represented 21 different biotypes and 22 different SmaI macrorestriction types. This observation confirmed that S. sciuri isolates obtained from insectivores and rodents living in natural environments constituted a heterogeneous population. Cluster analysis revealed that the macrorestriction patterns and the biochemical profiles matched in some cases. However, S. sciuri isolates that exhibited the same or closely related biochemical profiles were also found to be associated with different macrorestriction patterns. Analysis of the 29 S. sciuri isolates for their plasmid content and their sensitivity to antimicrobial agents showed that most of the isolates were plasmid‐free and susceptible to all antimicrobial agents tested.     

Verification by polymerase chain reaction of vertical transmission of Theileria sergenti in cows

To evaluate the transplacental transfer of Theileria sergenti infection in cattle, we used DNA probes to detect T. sergenti in 6 pregnant cows and their calves. All the animals were monitored by parasitologic, serologic, and polymerase chain reaction (PCR) assays for a predicted 875-base-pair (bp) DNA product and a 684-bp amplicon detected by nested PCR in the blood and spleens of aborted fetuses. An open reading frame (ORF) starting at nucleotide 170 and terminating at position 1021 was shown to code for a polypeptide of 283 amino acid residues. All 6 dams and 5 calves were positive for T. sergenti in all tests. One calf was positive only with nested PCR. We conclude that transplacental transmission of T. sergenti is a significant problem. The relevance of the data in the programmed introduction of new (especially pregnant) animals into established clean herds needs serious consideration with regard to control of theileriosis and other tickborne diseases.     

Manipulated Mouse Embryos as Bioassay System for Water Quality Control

Mouse pronuclear stage embryos with intact slit zona pellucida (manipulated) were cultured in vitro until the hatched blastocyst stage in simplex optimized medium with higher K⁺ concentration (KSOM) prepared with three different water types: tap, deionized reverse osmosis (D‐O) water and Milli‐Q system (M‐Q) water. The culture media were supplemented with or without protein and ethylenediaminetetraacetic acid (EDTA, disodium salt). The rates of hatched blastocysts were significantly affected (p < 0.01) by micromanipulation, protein supplement and water source. The water source has no influence (p > 0.05) on development in EDTA‐supplemented protein‐free culture media, whereas in EDTA‐ and protein‐free culture media, the water quality significantly (p < 0.001) affected the rates of development, with higher rates in media prepared with M‐Q water. The micromanipulated embryos showed higher sensitivity to the water quality (p < 0.01). It worth mentioning that the rates of hatched blastocysts in protein‐free culture media were very low (0–7.5%). Furthermore, the three different water types were analysed by measuring the electrical conductivity, inorganic ions, total organic carbon and endotoxins to evaluate the purity. M‐Q water showed the lowest levels of inorganic ion, total organic carbon and endotoxin concentrations. We concluded that manipulated mouse embryos are good system to evaluate the quality of water used in biological system.     

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.     

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.