Genomic variability among globally distributed isolates of equine arteritis virus.

Equine arteritis virus (EAV), a non-arthropod borne togavirus, has been shown to have a global distribution. To date, no major antigenic variation has been demonstrated between EAV isolates from different geographic origins. In this study, the genomic RNA of EAV isolates obtained from horses of different breeds in various countries around the world was oligonucleotide fingerprinted. Comparisons of these fingerprints were used to determine the extent of genomic variation among such isolates. Comparisons among isolates from North American horses revealed, for the most part, oligonucleotide homologies of less than 60%. Only 29 of the 98 comparisons revealed greater than 60% oligonucleotide homology. Nonetheless, several comparisons indicated a close epidemiologic relationship between isolates from horses of different breeds located in different states. Though all European isolates were of Standardbred origin and were from horses located in northern European countries, the majority had oligonucleotide homologies of less than 60%. Where oligonucleotide homology was apparent, it was, with one exception, greater than 70%. The two isolates from New Zealand had 93.2% oligonucleotide homology. This is indicative of an extremely close epidemiologic relationship. Comparisons between EAV isolates from around the world revealed oligonucleotide homologies between viruses from North America, Europe and New Zealand. In several instances, this homology was greater than 70% and in one case greater than 80%. No oligonucleotide homology was evident in comparisons involving the virus from South Africa. The high level of genomic conservation between certain EAV isolates of disparate geographic origins may reflect dissemination of the virus associated with the international movement of horses. The extent of genomic variation demonstrated between most of the EAV isolates used in this study confirms the need for further investigation of genomic heterogeneity among strains of this virus before techniques that rely upon nucleic acid hybridization can be effectively applied as diagnostic procedures.     

Toxic peripheral neuropathy with demyelination in Sprague-Dawley rats given CGS 21595--a 5-lipoxygenase inhibitor.

Male and female Sprague-Dawley rats were given CGS 21595, a pro-drug that is almost immediately metabolized to CGS 19213, a naphthoquinone that acts as a 5-lipoxygenase inhibitor. The compound was administered by gavage to five groups of Sprague-Dawley rats (group Nos. 1, 5, n = 30; group Nos. 2-4, n = 20) at daily doses of 0, 50, 150, 500, or 1,000 mg/kg for 13 weeks. Rats in the higher dose groups had a reduced weight gain, but significant neurologic signs were not observed. A peripheral neuropathy consisting predominantly of myelin destruction in the spinal nerve roots and sciatic nerves in male rats treated with greater than or equal to 150 mg/kg CGS 21595 and in female rats treated with greater than or equal to 50 mg/kg CGS 21595 for 13 weeks. This lesion was not fully reversible after a recovery period of 4 weeks. Lesions consisted of ballooning of myelin sheaths, infiltration by macrophages, demyelination, and occasional areas of remyelination. Axons were generally preserved, and the brain and spinal cord were not affected. Male and female rats in all treatment groups had cytoplasmic hyaline droplets in the proximal renal tubules. This change was reversible after 4 weeks and was not associated with any other adverse effects on the kidney.     

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.     

An evaluation of the pesera soil erosion model and its application to a case study in Zakynthos, Greece

Abstract. The Pan‐European Soil Erosion Risk Assessment (pesera ) model was evaluated using existing soil erosion data collected under various types of climate, vegetation, landscape and soil conditions. The data used represent a variety of typical Mediterranean land uses such as winter wheat, vines, olives and bare, stony land prevailing in hilly areas. Using this data, the model was calibrated for sediment transport by overland flow and results compared to measured soil erosion values from runoff plots and a watershed on a monthly basis. The performance of the model was assessed statistically, showing that it can be satisfactorily used for predicting soil erosion rates under the conditions included in the study. The overall model estimate including all the available experimental data was 0.69 t ha^?1 yr^?1 with a maximum error of 1.49 t ha^?1 yr^?1. After validation, the model was applied to a small watershed (60 ha) of great ecological importance for the sea turtle Caretta caretta. For this purpose, soil and vegetation maps were compiled from all the necessary data for applying the model. The model was run for three years using daily data from an existing nearby meteorological station. The predicted and measured soil erosion rates for a 7‐month period were 0.31 t and 0.18 t, respectively. Application of the model to each mapping unit showed the over‐riding importance of land use for sediment generation under the given climatic conditions. Bare land, occupying 5.5% of the watershed area, generated up to 69% of the total sediments estimated for the watershed. It is concluded that the pesera model can be used as a regional diagnostic tool under a range of soil, topographic and climatic conditions for identifying the best land use type and vegetation cover to protect hilly areas from soil erosion. The calculated overall root mean square error for the model is 0.06 t ha^?1 yr^?1, compared to a soil erosion rate of 0.04 t ha^?1 yr^?1, which can be tolerated for protecting the area for the sea turtle.     

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.     

Feeding Frequency Affects Growth, Not Fillet Composition, of Juvenile Sunshine Bass Morone chrysops×M. saxatilis Grown in Cages

Abstract.— In an effort to feed sunshine bass Morone chrysops × M. saxatilis efficiently, promote optimal growth, and reduce labor costs associated with feeding, sunshine bass were grown in cages and fed one of four feeding frequencies: once/d, twice/d, once every other day (I X/EOD), and twice every other day (2X/EOD) for 21 wk. Juvenile sunshine bass were fed a commercial floating diet containing 40% protein and 11.5% lipid. One hundred fish were hand-counted and stocked into each of 12 3.5-m³ cages with three replications per treatment. At the conclusion of the study, percentage weight gain of sunshine bass fed twice/d was significantly (P < 0.05) higher (1,850%) compared to fish fed all other feeding frequencies. Specific growth rate (SGR) of fish fed twice/d was significantly higher (2.1%/d) compared to fish fed all other feeding regimes, while fish fed once/d had a higher SGR (2.0%/d) compared to fish fed I X/EOD (1.6%/d) and 2X/EOD (1.8%/d). Percentage survival was not significantly different (P > 0.05) among all treatments and averaged 70.4%r. Feed conversion ratio (FCR) of fish fed twice/d was significantly higher (2.40) compared to fish fed all other feeding regimes. Percentage fillet weight of fish fed twice/d was significantly higher (27.8%) compared to all other treatments. Percentage moisture, protein, lipid, and ash in fillet were not significantly different among all treatments and averaged 75.7%, 19.4%, 3.5%, and 1.2%n, respectively (wet-weight basis). Based upon data from the present study, it appears that producers growing juvenile sunshine bass in cages may want to feed fish twice daily. This feeding regimen allows for higher growth rates, without adverse effects on body or fillet compositions. However, economic analysis needs to be conducted to determine if feeding twice/d is profitable.     

Effects of low and high calcium intake prepartum on calcium mobilization rate around parturition in dairy cows

Forty-one dairy cows were fed a low (LCa-13 g/d) and a high (HCa-83.5 g/d) calcium ration in the 8 weeks prior to parturition and the effect on the Ca mobilization rate around parturition was studied. Plasma Ca values were stable in the LCa group around parturition. In the older cows of the HCa group a very slight decrease in the mean plasma Ca was observed: 2.58 mmol/l at 12-36 h ante partum decreased to 2.38 mmol/l at parturition. Hypocalcaemia, which commonly occurs around parturition, did not occur in 40 of the cows. A subclinical hypocalcaemia (1.8 mmol/l) occurred in one cow (parity 10) from the HCa group. To assess the efficiency of Ca mobilization, a severe hypocalcaemia (1.0 mmol/l) with clinical signs was induced by means of Na2EDTA infusion (0.90 mmol/min), starting at 10 h post-partum. The older cows in the LCa group required more Na2EDTA than those in the HCa group. Higher urinary hydroxyproline excretion in the week before parturition in the LCa than in the HCa group suggested a higher bone turnover. Plasma PTH levels around parturition were not significantly different between the groups. The amount of colostrum milked out in the first 10 h post-partum did not influence Ca homeostasis around parturition. The results contradict those of many other experiments in which hypocalcaemia was observed in cows ingesting high levels of Ca. It is concluded that the restricted feed intake prepartum possibly had a favourable effect on Ca homeostasis.