Cryptosporidium infection in juvenile pet rabbits

Cryptosporidium infection was confirmed by fecal examination for the first time in pet rabbits in a wholesale store located in Kanagawa Prefecture, Japan. Fecal samples were obtained postmortem from juvenile rabbits (n=66), which had died after developing diarrhea. Feces from healthy rabbits (n=30) were also collected and examined as controls. Two types of Cryptosporidium oocysts distinctive in size and shape were found (Type A and B). Types A and B oocysts were detected from 16.7% and 13.6% of the diarrheic, and 3.3% and 0% of the normal feces, respectively. Since Cryptosporidium oocysts were detected at a higher rate in the diarrheic rabbits than in the healthy rabbits, special caution should be taken when handling a pet rabbit presenting with diarrhea.     

Effectiveness of small interfering RNA (siRNA) against the Mcl-1 gene in a canine mammary gland tumor cell line

The effect of down-regulation of Mcl-1 expression by small interfering RNA (siRNA) against the canine Mcl-1 gene on apoptosis was investigated by transfecting CF33 (canine mammary gland tumor cell line) with siRNA using cationic liposomes. The siRNA against canine Mcl-1 increased the rate of apoptotic cells and decreased the numbers of viable cells. Further, sequence-specific down-regulation of Mcl-1 expression was measured by real time-PCR and Western blot analysis. The siRNA directed against the Mcl-1 gene reduced both the mRNA and protein expression in the CF33. Our study suggests the importance of Mcl-1 in canine mammary tumors for inducing apoptosis and reinforces using Mcl-1 as a putative therapeutic target in canine mammary gland tumor.     

Detection of candidate polymorphisms around the QTL for fat area ratio to rib eye area on BTA7 using whole‐genome resequencing in Japanese Black cattle

In our previous study, we performed genome‐wide association study (GWAS) to identify the genomic region associated with Fat area ratio to rib eye area (FAR) and detected a candidate in BTA7 at 10–30 Mbp. The present study aims to comprehensively detect all polymorphisms in the candidate region using whole‐genome resequencing data. Based on whole‐genome resequencing of eight animals, we detected 127,090 polymorphisms within the region. Of these, 31,945 were located within the genes. We further narrowed the polymorphisms to 6,044 with more than five allele differences between the high and low FAR groups that were located within 179 genes. We subsequently investigated the functions of these genes and selected 170 polymorphisms in eight genes as possible candidate polymorphisms. We focused on SLC27A6 K81M as a putative candidate polymorphism. We genotyped the SNP in a Japanese Black population (n = 904) to investigate the effect on FAR. Analysis of variance revealed that SLC27A6 K81M had a lower p‐value (p = .0009) than the most significant SNP in GWAS (p = .0049). Although only SLC27A6 K81M was verified in the present study, subsequent verification of the remaining candidate genes and polymorphisms could lead to the identification of genes and polymorphisms responsible for FAR.     

A case of spontaneous Zymbal’s gland carcinoma with lung metastasis in an aged Fischer 344 rat

Zymbal’s gland neoplasms are induced in rats through the administration of various carcinogens, but spontaneous neoplasia is rare. This report describes a spontaneous Zymbal’s gland carcinoma with lung metastasis found in an aged male Fischer 344 rat. Macroscopically, the dome-like tumor nodule, approximately 30 mm in diameter with ulceration, was located near the ear canal of the rat. No healthy tissue or structure of Zymbal’s gland was identified on the corresponding side, while the normal salivary glands and a lacrimal gland were observed. Histologically, a large part of the tumor mass was occupied by poorly differentiated neoplastic cells, the shapes of which were oval to polygonal or fusiform. Additionally, clusters of sebaceous-like foamy cells and squamous metaplasia with prominent keratinization were observed. Tumor cells were found to metastasize to the lung; these cells displayed histological similarities, including a sebaceous gland-like pattern, to those in the primary site. The tumor cells were immunohistochemically positive for cytokeratin AE1/AE3 or vimentin but negative for CD68, S100, α-smooth muscle actin, von Willebrand factor, and desmin. Our results indicate that the tumor was a poorly differentiated Zymbal’s gland carcinoma with lung metastasis.     

Adjuvant-enhanced antibody responses in the absence of toll-like receptor signaling

Innate immune signals mediated by Toll-like receptors (TLRs) have been thought to contribute considerably to the antibody-enhancing effects of vaccine adjuvants. However, we report here that mice deficient in the critical signaling components for TLR mount robust antibody responses to T cell-dependent antigen given in four typical adjuvants: alum, Freund's complete adjuvant, Freund's incomplete adjuvant, and monophosphoryl-lipid A/trehalose dicorynomycolate adjuvant. We conclude that TLR signaling does not account for the action of classical adjuvants and does not fully explain the action of a strong adjuvant containing a TLR ligand. This may have important implications in the use and development of vaccine adjuvants.     

Growth behavior of suckers derived from transplanted sago palm (Metroxylon sagu Rottb.)

During the growth of sago palm (Metroxylon sagu Rottb.), primary suckers appear from the main stem (MS) of transplanted suckers. Then, secondary suckers appear from the primary suckers. After the MS (trunk) is first harvested 10 and several years after transplantation, trunks can be harvested persistently using the primary, secondary, and subsequent suckers, which are designated as derivative suckers, growing with the MS. However, little knowledge exists about the growth behavior of derivative suckers. This study clarified how derivative suckers, especially the primary and secondary suckers, spread in the horizontal direction, and how they form a plant with the MS during the creeping growth stage. Most derivative suckers crept in the direction of about 70° subtended by the mother stem. However, two primary suckers that appeared early after transplantation crept in an obtuse angle to the creeping direction of the MS. As the reason for this obtuse angle direction, we considered the following four factors: (1) the existence of petiole, (2) distance from the sucker to the ground surface, (3) enlargement of the MS, and (4) space for sucker growth. The growth behaviors of the two primary suckers and the other derivative suckers differed. Therefore, in sago palm cultivation, the two primary suckers which appeared first from the MS were very important for formation of the framework of the plant with the MS, in terms of efficient utilization of space for the growth of derivative suckers.     

F-wave latency and F-wave conduction velocity for the tibial nerve in clinically normal dogs

OBJECTIVE: To establish a method of F-wave examinations and to determine values of F-wave conduction velocity (FWCV) and F-wave latency for the tibial nerve of clinically normal dogs. ANIMALS: 21 clinically normal dogs. PROCEDURE: The F-waves were elicited from the interosseous muscles via stimulation of the tibial nerve. The FWCV was determined by using the F-wave shortest value and the surface distance corresponding to the tibial nerve length. Correlation between the smallest latency value of the F-wave and the length of the tibial nerve and between the FWCV and rectal temperature were closely examined. RESULTS: F-wave latency was proportional to the length of the tibial nerve (correlation coefficient, 0.929). Mean +/- SD FWCV was 77.98 +/- 8.62 m/s. Regression equation was as follows: F-wave latency = 2.799 + (0.029 X length of the tibial nerve).The FWCV was increased when the measured rectal temperature was high. Correlation coefficient between FWCV and rectal temperature was 0.665. CONCLUSIONS AND CLINICAL RELEVANCE: In the study reported here, we established a reliable method for clinical evaluation of the F-wave. When assessing nerve conduction velocity, it is essential to measure nerve length along the pathway that the nerve impulse travels. This method of F-wave examination is a useful diagnostic tool for the evaluation of suspected dysfunction of the peripheral nervous system.     

Screening of therapeutic targets for canine mast cell tumors from a variety of kinase molecules

Options of systemic treatment for canine MCT have been still limited and most canine cases with MCTs eventually undergo relapses even after achievement of a remission. Thus additional therapies are required to establish for the tumor. To identify the novel candidate therapeutic targets for canine MCT, the mRNA expression and phosphorylation statuses of several receptor or non-receptor kinases as well as the inhibitory effect of 95 specific inhibitors on the growth were assessed in three canine MCT cell lines (HRMC, VIMC1 and CMMC1). Among the 14 targets, the mRNAs of 11, 7 and 7 kinases were amplified in HRMC, VIMC1 and CMMC1, respectively. The mRNAs of VEGFR3, PDGFRα, SRC, YES, LCK and FYN were detected in all cell lines. The phosphorylation of 12, 8 and 7 kinases was observed by using specific antibody arrays in HRMC, VIMC1 and CMMC1, respectively. DTK, EPHB6, AMPKα1, CREB, STAT5a and STAT5b were phosphorylated in all cell lines. The 10, 9 and 17 inhibitors exhibited the biological activity against the growth of HRMC, VIMC1 and CMMC1, respectively. Only three inhibitors such as SB218078 (for Chk1), PDGF RTK inhibitor IV (for PDGFR) and radicicol (for Hsp90) suppressed the growth of all three cell lines. The present study indicated that several kinases, such as Chk1, PDGFR and Hsp90, could be used as therapeutic targets in the treatment for canine MCT. Further studies and clinical trials are warranted to apply the inhibitors for the treatment of the tumor.     

Comparison of two quantitative assays for xenotropic murine leukemia virus-related virus

Xenotropic murine leukemia virus-related virus (XMRV), a novel gammaretrovirus in humans, was found in patients with prostate cancer (PC) and chronic fatigue syndrome (CFS). However, there has been controversy whether XMRV is directly associated with human diseases. In this study, we developed a LacZ marker rescue assay using human embryonic kidney 293T cells and a focus assay using a feline fibroblastic sarcoma-positive leukemia-negative QN10S cells. XMRV induced prominent foci in QN10S cells and the viral titer determined by the focus assay was as high as that by the LacZ marker rescue assay. Because the focus assay is simple and sensitive, it will be useful for monitoring infectious XMRVs in CFS and PC patients and virological studies for XMRV.