Mammary lesions associated with bovine herpesvirus type 4 in a cow with clinical mastitis

Intranuclear eosinophilic inclusion bodies were seen in the lactiferous duct and sinus epithelium of mammary tissues collected from a cow with clinical mastitis. Transmission electron microscopy revealed herpesvirus particles in these cells. Immunolabeling against anti bovine herpesvirus type 4 (BHV-4) rabbit serum was detected in nuclei that had intranuclear inclusion bodies. In addition, BHV-4 was isolated from the mammary tissue. The viral DNA was detected by nested PCR from the same tissue. This is the first report to describe mammary lesions in association with BHV-4.     

Fibrinonecrotic rhinitis caused by a concurrent infection of Fusobacterium necrophorum and Arcanobacterium pyogenes in a cow

An 8 year-old cow showing severe dyspnea and nasal mucosal necrosis immediately after parturition was subjected to pathological examination. The principal lesions were fibrinonecrotic rhinitis, necrotic bronchopneumonia and renal infarction. Fusobacterium necrophorum biotype A and Arcanobacterium pyogenes antigens were detected in the nasal and pulmonary lesions. These results suggest that the lesions were caused by a concurrent infection of the detected bacteria and that the pulmonary lesions were caused by the aspiration of infectious materials from the nasal ones. Mucosal coagulative necroses observed as the initial lesions in rhinitis were frequently associated with multiple thrombosis. The findings might suggest that thrombosis played an important role in the development of the nasal lesions.     

Community structure of bacteria and fungi responsible for rice straw decomposition in a paddy field estimated by PCR-RFLP analysis

Rice straw including leaf sheaths and blades put in nylon mesh bags was placed in the plow layer of a Japanese paddy field after harvest under upland conditions and after transplanting of rice seedlings under flooded conditions. In addition, rice straw that was decomposed under the upland conditions during the off-crop season in winter was placed again in soil at the time of transplanting. The materials were collected periodically to analyze the community structure of the bacteria and fungi responsible for rice straw decomposition by PCR-RFLP analysis. The PCR products with 27f and 1492r primers designed for bacterial 16S rDNA and with EF3 and EF4 primers designed for fungal 18S rDNA were digested with four restriction endonucleases (Hinf I, Sau3A I, Hae III, EeoR I). Bacterial communities in the decomposing rice straw were different from each other between upland and flooded conditions, between leaf sheaths and blades, and between straw samples with and without decomposition under upland conditions during the off-crop season. Fungal communities in the decomposing rice straw were also different between the leaf sheaths and blades under upland soil conditions. Score plots of bacterial and fungal communities in the principal component analysis were separated from the plot of the straw materials along with the duration of the placement, indicating the succession of bacterial and fungal communities in decomposing rice straw with time.     

Effect of tissue deterioration on postmortem BSE diagnosis by immunobiochemical detection of an abnormal isoform of prion protein

Surveillance for bovine spongiform encephalopathy (BSE) in fallen stock in Japan is conducted with a commercial enzyme-linked immunosorbent assay (ELISA) for mass screening, with Western blotting (WB) and immunohistochemistry performed for confirmation of the ELISA. All tests are based on immunological detection of an abnormal isoform of the prion protein (PrP(Sc)) in brain tissues, which have sometimes deteriorated by the time samples from fallen stock reach a diagnostic laboratory. To evaluate BSE surveillance procedures for fallen stock, we examined PrP(Sc) detection from artificially deteriorated BSE-affected bovine brain tissues with a commercial ELISA kit and compared the results with those of WB. The optical density (OD) values of the ELISA decreased with advancing deterioration of the tissues, whereas no reduction in the signal for PrP(Sc) was observed in WB, even when performed after 4 days of incubation at 37 degrees C. The progressive decrease in the OD values in the ELISA appear to be caused by a partial loss of the N-terminal moiety of PrP(Sc) due to digestion by endogeneous and/or contaminated microbial enzymes, and by the presence of ELISA inhibitors that are generated in deteriorated tissues. These results suggest that WB is the most reliable test for fallen stock, especially for cattle brains within decaying carcasses.     

MRI examination of the masticatory muscles in the gray wolf (Canis lupus), with special reference to the M. temporalis.

We examined the head of the gray wolf (Canis lupus) using MRI methods. Although the arising surface of the M. temporalis was not so enlarged in the frontal bone, the small frontal bone did not disturb the M. temporalis from occupying the lateral space of the frontal area in the gray wolf as in the domesticated dog. In the gray wolf, it is suggested that the M. temporalis may not be well-developed in terms of size of arising area, but in the thickness of running bundles. We suggest that the dog has changed the three-dimensional plan of the M. temporalis during the domestication and that the M. temporalis has developed a large arising surface in the frontal bone and lost the thickness of belly in the frontal area in accordance with the enlargement of the frontal bone and the increase in brain size.     

PCR-DGGE analysis of bacterial communities in <Emphasis Type="Italic">funazushi</Emphasis>, fermented crucian carp with rice,during fermentation

Funazushi (fermented Crucian Carp with rice) is a fermented fish product found only around Lake Biwa in Shiga Prefecture, Japan. It is characterized by a unique cheese-like flavor and characteristic sour taste. We analyzed the changes in the microbial community during funazushi fermentation by denaturing gradient gel electrophoresis of PCR-amplified 16S rDNA fragments (PCR-DGGE) and by plate counts. The plate counts showed that lactic acid bacteria reached 8.0 log₁₀ CFU/g within 7 days of fermentation initiation before decreasing slowly to 4.0 log₁₀ CFU/g during the remainder of 1-year study period. PCR-DGGE revealed that the dominant bacteria in the initial (days 14 and 30) and latter (days 90, 180, and 360) periods of fermentation were Lactobacillus plantarum and L. acetotolerans. This is the first identification of L. acetotolerans in funazushi as traditional cultivation techniques have not been sufficiently sensitive. This is the first report of PCR-DGGE being used to assess the microbial community in funazushi. This technique was also found to be effective in profiling microbial diversity.     

Suppression of Ralstonia solanacearum in soil following colonization by other strains of R. solanacearum

The effect of prior colonization of a sterile loam soil and a sterile clay loam soil by individual soil bacteria on the subsequent growth of a bacterial wilt pathogen Ralstonia solanacearum YU1Rif43 (tRNA type III: Seal et al. 1992: Appl. Environ. Microbiol., 58, 3759–3761) was investigated. Various strains, belonging to the same type, the same species, the same genus, Gram-negative, Grampositive, or fungi, were used. The degree of suppression of the growth of R. solanacearum YU1Rif43 was markedly different depending on the species that had previously colonized the soil, hereafter referred to as priorcolonists. All the strains belonging to R. solanacearum type III suppressed the growth of R. solanacearum YU1Rif43 markedly, while strains of R. solanacearum type I and type II showed a moderate suppressive effect on R. solanacearum YU1Rif43. The suppressive effect of the strains belonging to species other than R. solanacearum, including fungal strains, was relatively limited, or some strains did not show any suppressive effect. The production of bacteriocin did not appear to be related to the strong suppressive effect of the R. solanacearum type III strains. Possible mechanisms for the suppressive effect of priorcolonists on R. solanacearum YU1Rif43 are discussed in relation to nutrients and physical sites in soil available for growth.     

Sites and processes of nutrient accumulation in the subsoil through water percolation from the plow layer in a submerged paddy soil microcosm

The leaching of nutrients from the plow layer by water percolation and their accumulation in the subsoil observed in a Japanese paddy field (Katoh et al. 2004: Soil Sci. Plant Nutr., 50, 721-729) were determined semi-quantitatively in a soil column experiment. Ca²⁺, Mg²⁺, K⁺, Mn²⁺, Fe²⁺, and phosphate in percolating water from the plow layer soil column were retained in the subsoil columns that were connected to the plow layer soil column. Fe²⁺, K⁺, and phosphate accumulated in the uppermost part of the subsoil. Accumulation of Fe²⁺ in the uppermost part of the subsoil was presmnably due to the cation exchange process with concomitant desorption of Ca²⁺. In contrast, Ca²⁺ and Mg²⁺ in percolating water from the plow layer soil colmnn accumulated once in the subsoil, and translocated downwards slowly with successive water percolation. Considerable amounts of inorganic carbon (IC) and dissolved organic carbon (DOC) in percolating water from the plow layer soil column were also retained in the subsoil columns. IC did not accumulate a gaseous form.