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.     

THE H-ION CONCENTRATION AND THE ORIGIN OF THE HEART BEAT

The rate of the frog or guinea pig heart preparation was not significantly influenced by the H-ion concentration when this was kept within limits which allowed the heart to continue beating. The theory that the H-ion concentration is intimately concerned with the origination of the heart beat is not supported by the results of this investigation.     

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).