First report of fig mosaic virus infecting common fig (Ficus carica) in Japan

For the first time, fig mosaic virus (FMV) was detected in common fig (Ficus carica) trees in Shimane, Japan. These trees exhibited mosaic, ringspots, or distortion, accompanied by chlorosis on leaves and yellow spots on fruits. Some of the symptomatic trees were infested with the eriophyid mite Aceria ficus. The virus was detected based on RT-PCR, followed by sequencing. The amplified 300 base-pair fragments shared 83.5–91.5% identity with the corresponding region of RNA-dependent RNA polymerase gene of FMV isolates previously reported in Turkey, Iran, and Italy.     

Calreticulin expression in neoplastic versus normal dog mammary glands: a cDNA subtraction-based study

This is the first report describing the expression of canine calreticulin (cCRT) in canine mammary gland tumour (MGT). Using cDNA subtraction method, it is found that mRNAs of CRT, cathepsin A, ovostatin, and lactotransferrin were differentially expressed in mammary adenocarcinoma as compared to hyperplasia, both of which were obtained from the dog. Furthermore, the mRNA expression levels of CRT and cathepsin A were significantly higher in canine MGT samples than in nontumour samples. In contrast, immunohistochemical studies have indicated that the expression of cCRT protein found to be detected in most of mammary gland tissues and was not correlated to the types of canine MGTs. Furthermore, cCRT was molecularly cloned, and the amino acid sequence of cCRT was found to be very similar to those of other species. Further studies are required to elucidate additional roles of cCRT in canine MGT.     

Significance of tumor-infiltrating immune cells in spontaneous canine mammary gland tumor: 140 cases

The numbers of tumor infiltrating T lymphocytes, B lymphocytes and antigen presenting cells were evaluated in an immunohistochemical manner in 140 canine spontaneous mammary gland tumor (MGT) tissues. As a result, we found a statistically significant increase in the number of intratumoral T lymphocytes (23.2 ± 23.8) in the malignant MGT group (n=51) compared with the benign MGT group (14.0 ± 16.0, n=89; P<0.05). Moreover, the high T lymphocyte infiltration in the malignant group correlated with poor prognosis in multivariate analysis (P<0.05). This study indicated the relationship between increased infiltrating T lymphocytes and canine MGT malignancy.     

Comparison of behavioral effects of morphine and fentanyl in dogs and cats

Behavioral effects induced by intravenous administration of morphine at 0.3, 0.6, 1.2, and 2.4 μg/kg and fentanyl at 5, 10, 20, and 40 μg/kg were evaluated in dogs and cats. In dogs, fentanyl and morphine depressed activity and level of consciousness in a dose- dependant manner. In cats, higher doses of fentanyl stimulated activity temporarily, but excitement, so-called "opioid mania," was not observed. Morphine induced distinctive behavioral changes characterized by sitting with fixed staring, and "opioid mania" was not observed in cats.     

Correlation between villous height and the disaccharidase activity in the small intestine of piglets from nursing to growing

Early weaning induces villous atrophy in the small intestine. Reduction in villous height in the small intestine after weaning is associated with reductions in brush‐border enzyme activity. Body weight gain after weaning is, therefore, correlated with villous height. This evidence suggested that the maintenance of small intestinal structure and function after weaning is important for the growth of young pigs. On the other hand, the relationship between villous height and the activity of the digestive enzymes in the small intestine has not been studied with piglets from the suckling to the growing period. Five suckling piglets, four piglets in the proximal stage of weaning, four pigs in the distal stage of weaning and four growing pigs were used. The activities of lactase (LA), sucrase (SA) and maltase (MA) were determined. LA showed a positive correlation with villous height in weaning. SA and MA were positively correlated with villous height from suckling to growing. In a previous study, non‐infectious dyspeptic diarrhea was frequently observed in growing piglets on Japanese swine farms. The maintenance of villous height to retain disaccharidase activity may prevent dyspepsic diarrhea in this stage.     

Ozone treatment of spent medium from Auricularia polytricha cultivation for enzymatic saccharification and subsequent ethanol production

We examined the enzymatic saccharification and ethanol fermentation of the spent media (SMs) from Auricularia polytricha cultivation using wood meals of Falcataria moluccana, Shorea sp., and Tectona grandis. Although the hydrolysis weight decrease and reducing sugar yield were higher in SM of F. moluccana, the ethanol yield was higher in SM of Shorea sp. Ozone treatment of SM further increased the hydrolysis weight decrease, reducing sugar, and ethanol yields in Shorea sp. These results indicate that SM of A. polytricha is a suitable biomass material to produce fermentable sugars for ethanol production, and that ozone treatment is a suitable method for increasing the ethanol yield.     

Improvement of DNA extraction method for dried blood spots and comparison of four PCR methods for detection of Babesia gibsoni (Asian genotype) infection in canine blood samples

To eradicate canine babesiosis in epidemic areas, mass-screening of the infection situation of Babesia gibsoni including occult infection is necessary. The development of cost-effective method for storage and transport of blood samples is required. A highly efficient DNA extraction procedure from dried blood spots (DBS) onto Whatman 3MM filter paper was developed for the diagnosis of B. gibsoni infection in dog by PCR. In 3 extraction methods, Chelex-based method in combination with saponin washing and phenol-chloroform-isoamyl alcohol extraction (Saponin-PCI method) provided the best results. Sensitivity of the 4 previously described PCR methods for detection of B. gibsoni infection was also compared using serially diluted blood samples of B. gibsoni-infected dogs. The PCR method using Gib599F/Gib1270R primer pair provided the best performance. To evaluate the stability of DNA in DBS, DBS of B. gibsoni-infected dogs stored at room temperature for 2 months. The stability was superior to whole blood samples stored at -20 degrees C for 2 months. This highly efficient DNA extraction method on DBS using Whatman 3MM filter paper has potential to be cost-effective and high performance tool for storage, and molecular diagnosis of clinical blood sample from dog. This procedure in combination with the PCR method using Gib599F/Gib1270R primer pair may greatly assist in diagnosis of B. gibsoni infection in dog populations that are geographically distant.     

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.