Genomic gene encoding manganese peroxidase from a white-rot fungus Phanerochaete crassa WD1694

The gene encoding manganese peroxidase of a white-rot fungus Phanerochaete crassa WD1694 was cloned and sequenced. Four genomic clones were sequenced in which 3 clones were existed as alleles. The analysis of intron–exon structures divided the 4 clones into three subfamilies that corresponded to mnp2 and mnp3 of Phanerochaete chrysosporium, and a new subfamily possessing only five introns. The purified P. crassa WD1694 MnP consisted of 4 isozymes with same molecular weight, same N-terminal sequence, and different pI. N-terminal sequence of deduced protein of P. crassa mnpB3 gene was identical to those of 4 MnP isozymes; however, the other 3 mnp genes had different N-terminal sequence. The molecular weight of encoded mature protein of mnpB3 gene and purified MnP had a gap that could be difference between MnP proteins encoded by single gene. The results suggested that 4 MnP isozymes of P. crassa WD1694 arose from single gene.     

Assessment of the decay risk of airborne wood-decay fungi III: decay risks at different sampling sites

The decay risk of airborne wood-decay fungi in the same volume of air was investigated by using an air sampler over the course of a year at three different sampling sites. Japanese cedar disks measuring 7.8 cm in diameter and about 3 mm in thickness, and with a moisture content of about 100 % were placed in a “BIOSAMP” air sampler and then exposed to 1000 l of air in the northern, central, and southwest parts of Japan. The exposed disks were incubated for 20 weeks in a damp container maintained at 26 ± 2 °C and degraded by fungi trapped on the disks. The decay risk was calculated from the mass loss during incubation, and the factors affecting the said risk were explored. The results showed that sampling sites apparently do not affect decay risk, even though the Scheffer’s climate indexes of the sites were quite different. The relation between the sampling month and decay risk reveals that decay risk remains virtually the same year-round. Relative humidity on a sampling day is one of the key factors affecting decay risk in sampling conducted at the central or southwest site. In contrast, no weather factors influenced decay risk at the northern sampling site.     

Physiological responses of young thoroughbred horses to intermittent high-intensity treadmill training

Background

Training of young Thoroughbred horses must balance development of cardiopulmonary function and aerobic capacity with loading of the musculoskeletal system that can potentially cause structural damage and/or lameness. High-speed equine treadmills are sometimes used to supplement exercise on a track in the training of young Thoroughbreds because the horse can run at high speeds but without the added weight of a rider. We tested the hypothesis that intermittent high-intensity exercise on a treadmill of young Thoroughbred horses entering training can enhance development of aerobic capacity (Vo₂max) and running performance more than conventional training under saddle, and do so without causing lameness.

Results

Twelve yearling Thoroughbreds trained for 8 months with conventional riding (C) only, conventional riding plus a short (2 month, S) interval of once-per-week high-intensity treadmill exercise, or a long (8 month, L) interval of once-per-week high-intensity treadmill exercise. Three treadmill exercise tests evaluated Vo₂max, oxygen transport and running performance variables in June of the yearling year (only for L), October of the yearling year and April of the 2-year-old year. No horses experienced lameness during the study. Aerobic capacity increased in all groups after training. In both October and April, Vo₂max in L was higher than in C, but did not differ between L and S or S and C. Running speeds eliciting Vo₂max also increased in all groups after training, with S (809 ± 3 m/s) and L (804 ± 9 m/s) higher than C (764 ± 27 m/s). Maximum heart rate decreased for all groups after training. Hematocrit and hemoglobin concentration increased for L throughout training.

Conclusions

Young Thoroughbred horses can increase aerobic capacity and running performance more than by strictly using track training under saddle with the addition of intermittent high-intensity treadmill exercise, and they can do so without experiencing lameness. This finding suggests that young racehorses might be able to achieve higher aerobic fitness during training without subjecting their musculoskeletal systems to increased loading and risk of developing lameness. The findings of this preliminary study do not indicate a specific protocol to best achieve this goal.     

Loop-mediated isothermal amplification for the rapid, sensitive, and specific detection of the O9 group of Salmonella in chickens

In the present study, the loop-mediated isothermal amplification (LAMP) assay was developed to amplify the fragments of the O9 Salmonella-specific insertion element and evaluated in the laboratory for its potential use in a field situation, such as poultry farms. Among the bacteria tested, a positive reaction was observed only for 128 strains of 6 serovars of the O9 group Salmonella, such as Enteritidis (SE) and Pullorum. The detection limit of the LAMP assay was 10(3)CFU/ml, which was more sensitive than that of the polymerase chain reaction (PCR) assay with the same target gene (10(6)CFU/ml). The final results were obtained within 30 min for the LAMP assay, while the PCR assay needed a total of 120 min. When the LAMP assay was applied to the enrichment broth mixed with cecal dropping samples either spiked with SE in vitro or excreted by SE-inoculated hens, the results were comparable to those of the conventional plating method including 2 separate enrichments. In conclusion, the LAMP assay developed in the present study is an effective method for the specific detection of the O9 group Salmonella serovars, including SE.     

Hypereosinophilic syndrome in two cats

Two cats showing chronic vomiting, diarrhea and weight loss were found to have leukocytosis with marked eosinophilia. Both cats were diagnosed with hypereosinophilic syndrome by the findings of increased eosinophils and their precursors in the bone marrow, eosinophilic infiltration into multiple organs, and exclusion of other causes for eosinophilia. Although cytoreductive chemotherapy with hydroxycarbamide and prednisolone was performed, these two cats died 48 days and 91 days after the initial presentation.     

Identification of c-kit mutations-independent neoplastic cell proliferation of canine mast cells

Gain-of-function mutations in the proto-oncogene c-kit have been considered the molecular mechanism of neoplastic proliferation of mast cells. However, the importance of c-kit gene mutations is not well evaluated in canine mast cell tumors (MCTs). In the present study, we established and characterized a mast cell line, HRMC, derived from a dog with MCT. We also examined c-kit mutations in HRMC cells and assessed an inhibitory effect of a tyrosine kinase inhibitor, STI571, on HRMC cells. HRMC cells had cytoplasmic metachromatic granules, chymase and tryptase, and expressed both KIT and FcepsilonRI on the cell surface. HRMC cells contained histamine and released beta-hexosaminidase through FcepsilonRI cross-linking and calcium ionophore stimulation. Nucleotide sequence analysis demonstrated no mutations in an open reading frame of c-kit cDNA and genomic DNA of the juxtamembrane domain of c-kit in HRMC cells. STI571 did not show any inhibitory effects on the proliferation of HRMC cells. These findings clearly demonstrated the existence of c-kit mutations-independent neoplastic canine mast cell proliferation. The growth factor-independent mast cell line established in this study might be valuable to explore novel mechanisms of c-kit mutations-independent neoplastic proliferation of mast cells in dogs.     

Molecular pathogenesis of feline leukemia virus-induced malignancies: insertional mutagenesis

Feline leukemia virus (FeLV), which is subclassified into three subgroups of A, B and C, is a pathogenic retrovirus in cats. FeLV-A is minimally pathogenic, FeLV-C can cause pure red cell aplasia, and FeLV-B is associated with a variety of pathogenic properties such as lymphoma, leukemia and anemia. FeLV-induced neoplasms are caused, at least in part, by somatically acquired insertional mutagenesis in which the integrated provirus may activate a proto-oncogene or disrupt a tumor suppressor gene. The common integration sites for FeLV have been identified in six loci with feline lymphomas: c-myc, flvi-1, flvi-2 (contains bmi-1), fit-1, pim-1 and flit-1. Oncogenic association of the loci includes that c-myc is known as a proto-oncogene, bmi-1 and pim-1 have been recognized as myc-collaborators, fit-1 appears to be closely linked to myb, and flit-1 insertion is shown to be associated with over-expression of a cellular gene, e.g. ACVRL1. Thus, identification of common integration sites for FeLV is a tenable model to clarify oncogenesis. Recent advances in molecular biology and cytogenetics have developed to rapidly detect numbers of retroviral integration sites by genome-wide large-scale analyses. Especially, polymerase chain reaction (PCR)-based strategies and chromosome analyses with fluorescence in situ hybridization (FISH) will be applicable for studies on FeLV.     

Parent‐of‐origin effects on carcass traits in Japanese Black cattle

Variances caused by the differential expression of paternally and maternally imprinted genes controlling carcass traits in Japanese Black cattle were estimated in this study. Data on marbling score (BMS), carcass weight, rib thickness, rib‐eye area (REA) and subcutaneous fat thickness (SFT) were collected from a total of 13,115 feedlot steers and heifers in a commercial population. A sire–maternal grandsire model was used to analyse the data, and then, imprinting parameters were derived by replacing the genetic effect of the dam with the effect of the maternal grandsire in the imprinting model to calculate the genetic parameter estimates. The proportions of the total genetic variance attributable to imprinted genes ranged from 8.7% (SFT) to 35.2% (BMS). The remarkably large imprinting variance of BMS was mainly contributed by maternally expressed inheritance because the maternal contribution of the trait was much larger than that of the paternal trait. The parent‐of‐origin effect originating from maternal gene expression was also observed for REA. The results suggested the existence of genomic imprinting effects on the traits of the Japanese Black cattle. Hence, the parent‐of‐origin effect should be considered for the genetic evaluation of Japanese Black cattle in breeding programmes.