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.     

Characterisation of cross-reactivity of virus neutralising antibodies induced by feline panleukopenia virus and canine parvoviruses

It was recently reported that canine parvoviruses (CPV) had entered cat populations and induced disease in infected cats, while they had affected only dogs in the past. It is important to determine whether conventional feline panleukopenia virus (FPLV) vaccines protect against recent CPV infections. In this study, the cross-reactivity of virus-neutralising (VN) and haemagglutinin-inhibition (HI) antibodies in cats induced by FPLV and CPV s were examined. Lower cross-reactivities of VN and HI antibodies against each CPV strain were observed in cats experimentally inoculated with FPLV or vaccinated with an inactivated FPLV vaccine. In addition, we revealed the existence of a novel type of FPLV, which reacted weakly with antibodies induced by the conventional FPLV vaccine.     

Oxidative damage to the membrane of canine erythrocytes with inherited high Na, K-ATPase activity.

Oxidative damage to the membrane in canine erythrocytes with inherited high Na, K-ATPase activity (HK cells) was compared with that in normal canine cells (LK cells). When 30 mM beta-acetylphenylhydrazine (APH) was applied to HK and LK cells, lipid peroxidation and hemoglobin denaturation occurred. Lipid peroxidation determined from malondialdehyde (MDA) formation was significantly lower in HK than in LK cells so far as endogenous glutathione (GSH) concentration was maintained at appropriate levels. With the depletion of GSH, MDA formation was accelerated and difference between HK and LK cells was not significant. Denatured hemoglobin bound to the membrane protein was less in HK than in LK cells. During incubation with APH, osmotic fragility increased markedly in LK cells, while HK cells showed very little change. The amounts of total lipid, total and free cholesterol, glycolipid, phospholipid and fatty acids were essentially the same in both cell types. Fatty acid compositions showed very small differences. The membrane of HK cells thus appear to have greater protection against oxidative damage induced by APH, owing to the presence of excess GSH in HK cells. The capability of HK cells to withstand oxidative damage would not be due to differences in membrane lipid compositions.     

A cat with acute myeloblastic leukemia without maturation (M1) treated with combination chemotherapy

A 2-year-old domestic shorthair cat was presented to us with decreased activity and anorexia. Hematologic findings revealed a mild non-regenerative anemia, thrombocytopenia, and leukocytosis with an increase in blast cells. Bone marrow aspirates also revealed a marked increase of blasts. The blastic cells were shown to be positive for peroxidase. Acute myeloblastic leukemia without maturation (M1) was diagnosed according to the FAB classification. Chemotherapy was initiated with cyclophosphamide, vincristine, prednisolone, and cytosine arabinoside. The cat responded partially. In total, the cats were given 7 blood transfusions. The cat died 14 weeks after first being presented to us.     

Enzymatic profiles of Erysipelothrix rhusiopathiae and Erysipelothrix tonsillae

The enzymatic activities of 39 strains of Erysipelothrix rhusiopathiae and 34 of E tonsillae were determined with the API ZYM system. The profiles of these two species were very similar, differing solely in N-acetyl-beta-glucosaminidase activity. Whereas 90 per cent of strains of E rhusiopathiae exhibited strong activity with N-acetyl-beta-glucosaminidase, positive reactions were observed for this enzyme in only 24 per cent of strains of E tonsillae. These results support previous DNA-DNA hybridisation studies and suggest that E tonsillae is a new species of the genus Erysipelothrix.     

Antiserum against culture filtrate is cross-protective for various serovars of Erysipelothrix rhusiopathiae

The protective effect of porcine antiserum prepared against culture filtrate (CF) of an attenuated strain of Erysipelothrix rhusiopathiae (serovar 2) in mice to challenge with 20 virulent strains of 18 serovars and one type N was investigated. Passively immunized mice survived after challenge with serovars 1a, 1b, 2, 5, 6, 8 (strain Goda), 11, 12, 15, 16, 21 or type N, but 10-30% mortality occurred in immunized mice challenged with each strain of serovars 4, 7, 8 (strain 911), 9, 18 or 19 and 70% mortality to serovar 10 (strain 2179). All immunized mice died after challenge with serovar 20 (strain 2553). Non-treated control mice died after challenge with all serovars and the type tested.     

Correlation between adherence of Erysipelothrix rhusiopathiae strains of serovar 1a to tissue culture cells originated from porcine kidney and their pathogenicity in mice and swine

Adherence of four virulent and four avirulent strains of Erysipelothrix rhusiopathiae, serovar 1a, to porcine kidney cell lines, PK-15 and ESK cells, was examined in an in vitro system. The virulent strains adhered well to the cells (range of means, 9.95 +/- 0.87-36.01 +/- 1.10 per cell). In contrast, the avirulent strains showed negligible adherence to the cells (range of means, 0.11 +/- 0.04-1.41 +/- 0.13 per cell). Pretreatment of bacteria with heat, trypsin, or antiserum resulted in a marked decrease in adherence. Scanning electron microscopic examination revealed that the bacteria attached directly to the microvilli of cells.