Journal of Veterinary Medical Science
Online ISSN : 1347-7439
Print ISSN : 0916-7250
ISSN-L : 0916-7250
Volume 76, Issue 1
January
Displaying 1-25 of 25 articles from this issue
Anatomy
  • Sadullah BAHAR, Durmus BOLAT, Mustafa Orhun DAYAN, Yahya PAKSOY
    2014 Volume 76 Issue 1 Pages 37-44
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 05, 2013
    JOURNAL FREE ACCESS
    The 2- and 3-dimensional (3D) anatomy and the morphometric properties of the paranasal sinuses of the foal have received little or no attention in the literature. The aim of this study was to obtain details of the paranasal sinuses using multiplane CT imaging to create 3D models and to determine morphological and morphometric data for the sinuses using the 3D models. The heads of five female foals were used in this study. The heads were scanned using computed tomography (CT) in the rostrocaudal direction. After the heads had been frozen, anatomical sections were obtained in the scan position. The 3D models of sinuses and the skull were prepared using MIMICS®. These models were used to assess the surface area and volume of the sinuses, the width, height and orientation of the apertures connecting these sinuses and finally the planar relation of the sinuses with the skull. The right and left sides of all anatomical structures, except the sphenoid sinuses, had symmetric organization on CT images and anatomical sections. The total sinus surface area and volume on both sides were 214.4 cm2 and 72.9 ml, respectively. The largest and the smallest sinuses were the frontal sinus (41.5 ml) and the middle conchal sinus (0.2 ml), respectively. It was found that the planes bounding the sinuses passed through easily palpable points on the head. In conclusion, 3D modeling in combination with conventional sectional imaging of the paranasal sinuses of the foal may help anatomists, radiologists, clinicians and veterinary students.
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  • Naoki TSUKAHARA, Yuri TANI, Hideyuki KIKUCHI, Shoei SUGITA
    2014 Volume 76 Issue 1 Pages 93-95
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 26, 2013
    JOURNAL FREE ACCESS
    Differences in the ultraviolet (UV) cutoff of ocular media between birds and mammals have been revealed by spectrophotometric measurements of the transmission of light wavelengths by the cornea, lens and vitreous body in chickens, crows, quails, rats, rabbits and pigs. The light transmission values of the cornea were shown to be above 50% for wavelengths of 330–800 nm in birds, 300–800 nm in rat and 310–800 nm in mammals except for rat. For the lens, the light transmission values were shown to be above 50% for wavelengths of 320–800 nm in birds and rat and 390–800 nm in mammals except for rat. Thus, among the ocular media, the cornea in birds and the lens in mammals except for rat may play a role as a major UV cutoff filter.
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Avian Pathology
  • Fumiya KAWAHARA, Guohong ZHANG, Takayuki SUZUKI, Akira IWATA, Kisaburo ...
    2014 Volume 76 Issue 1 Pages 25-29
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 21, 2013
    JOURNAL FREE ACCESS
    None of anticoccidial vaccines (Trivalent TAM™, monovalent Neca™ and imported pentavalent Paracox®-5) contain Eimeria brunetti in Japan, which has not been regarded as a cause of coccidiosis, because of its low prevalence. However, we have recently reported the evidence of a high nationwide prevalence of this species. In this report, we describe the characteristics of E. brunetti which have never been clearly defined in Japan. Mortality rates and other disease characteristics caused by our strain (Nb strain) were similar to those reported previously in other studies. Despite great reduction of body weight gains among groups infected with over 1 × 103 oocysts, the intestinal lesions in the infected chickens were rather mild compared to previous studies. Sulfa drugs and lasalocid were so effective that the E. brunetti infection was almost completely blocked. Consequently, it is suggested that the characteristics of E. brunetti are various among the strains, but the pathogenicity of the Japanese Nb strain is enough strong to cause clinical coccidiosis.
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  • Kikuyasu NAKAMURA, Mitsuru ITO, Toshiki NAKAMURA, Yu YAMAMOTO, Manabu ...
    2014 Volume 76 Issue 1 Pages 31-36
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 22, 2013
    JOURNAL FREE ACCESS
    The pathogenicity of Newcastle disease (ND) virus, isolated from ND outbreak in vaccinated chickens, was evaluated through experiments. The pathogenicity indexes (mean death time (MDT); 58 hr, intracerebral pathogenicity index (ICPI); 1.7 and intravenous pathogenicity index (IVPI); 2.51) indicated that the ND virus was velogenic. The ND virus caused lymphocytic necrosis in the spleen with fibrinous exudation and proliferation of macrophages, sinusoidal fibrin exudation in the liver, proliferation of macrophages in the lung, lymphocytic necrosis and depletion in the bursa of Fabricius, cecal tonsils and thymus, necrosis of bone marrow, tracheitis, conjunctivitis and necrosis of feather epithelial cells in specific-pathogen-free chickens. Immunohistochemically, ND virus antigens were seen in the lesions mentioned above. The ND virus could not induce the encephalitis and pancreatitis that were observed in the natural case of ND in vaccinated chickens. There was no clinical disease in vaccinated chickens after the challenge of the ND virus. In diluted ND vaccine experiments, chickens vaccinated with a high dilution of vaccine and then challenged with the ND virus showed clinical sign and mortality with pancreatic focal necrosis. Vaccine diluted with fresh tap water had no effect on protection against the challenge of the ND virus. This study suggests that improper vaccination may be involved in outbreaks of ND in vaccinated chickens.
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  • Seiko FURUKAWA, Kenji TSUKAMOTO, Minoru MAEDA
    2014 Volume 76 Issue 1 Pages 89-92
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 23, 2013
    JOURNAL FREE ACCESS
    Gross lesions characterized by swollen livers and spleens accompanied by diffuse white miliary spots, which resembled those of Marek’s disease, were detected in two flocks of local meat-type chickens at a Japanese poultry processing plant in June and August 2010. The microscopic examinations revealed proliferative foci consisting of spindle or polymorphic cells in the interstitium of livers, splenic follicles and the interstitium of kidneys. These cells were positive immunohistochemically with Iba1 antibody, indicating they were histiocytic cells. Some of them contained antigens of avian leukosis virus (ALV) by immunohistochemistry,and the env gene of ALV subgroup J was detected from the spleens by polymerase chain reaction (PCR). Phylogenetic analysis of the PCR product indicated that the env gene might be descended from the American ADOL-7501 strain of ALV-J. These results suggest that the swollen livers and spleens of the meat-type chickens may come from histiocytic proliferation caused by ALV-J infection.
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Bacteriology
  • Satoko KAWAJI, Reiko NAGATA, Yasuyuki MORI
    2014 Volume 76 Issue 1 Pages 65-72
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 20, 2013
    JOURNAL FREE ACCESS
    An efficient protocol for the manual fluorescent MGIT culture system combined with rapid confirmation of Mycobacterium avium subsp. paratuberculosis (MAP) growth in the broth culture was established and evaluated for the detection of viable MAP in direct quantitative PCR (QPCR) positive bovine feces. Manually detected fluorescence emissions from MGIT tubes were analyzed objectively using an open source software, ImageJ. For molecular confirmation of MAP growth, DNA samples harvested by simply boiling the broth, an inexpensive and time- and labor-saving DNA preparation method, yielded adequate results. The sheep strain of MAP required longer incubation time relative to the cattle strain, suggesting that the MGIT system may not support well the growth of ovine isolates as described previously. Of 61 direct QPCR positive bovine feces, the recovery rate of MAP in the MGIT system (62.3%) was significantly higher (P<0.05) than that using 7H10 agar-based slants (44.3%). The time to obtain a final result for fecal culture by the MGIT system was several weeks earlier compared to solid media. In MGIT culture positive samples, the time to detect fluorescence was correlated with the DNA quantity detected in fecal QPCR. As a positive result in the direct fecal QPCR test does not mean fecal excretion of viable MAP, bacterial isolation by fecal culture could be conducted to verify the QPCR result. For this purpose, the manual MGIT system is a sensitive and rapid culture method at least for bovine samples.
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Biochemistry
  • Hiroki ASANO, Yohei KANAMORI, Satoshi HIGURASHI, Takayuki NARA, Ken KA ...
    2014 Volume 76 Issue 1 Pages 57-64
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 20, 2013
    JOURNAL FREE ACCESS
    There are two types of brown adipocytes: classical brown adipocytes that form the brown fat depots and beige adipocytes that emerge in the white fat depots. Beige adipocytes have a low level of uncoupling protein 1 (Ucp1) expression in the basal state, but Ucp1 expression is increased in response to β adrenergic receptor activation. The present study explored the factors responsible for the differentiation of 3T3-L1 white preadipocytes to beige adipocytes. Significant expression of Ucp1 was not detected under any tested conditions in the absence of isoproterenol (Iso), an agonist of β adrenergic receptor. Iso-induced Ucp1 expression was significantly higher in the cells treated with a mixture of triiodothyronine (T3) and 3-isobutyl-1-methylxanthine (IBMX) for days 0–8 than in the control cells. Chronic IBMX treatment was indispensable for the enhanced Iso-induced Ucp1 expression, and treatment with additional rosiglitazone (Rosi) for days 0–8 further increased the Ucp1 expression. Recently, genes were identified that are predominantly expressed in beige adipocytes, which were induced from stromal vascular cells in white fat depots. However, the expression levels of the beige adipocyte-selective genes in the adipocytes induced by the mixture of T3, IBMX and Rosi did not differ from those in the control adipocytes. The present study indicates that 3T3-L1 cells can differentiate to beige-like adipocytes by prolonged treatment with the mixture of T3, IBMX and Rosi and that the gene expression profile of the adipocytes is distinct from those previously induced from white fat depots.
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  • Masashi WATANABE, Kazuaki TANAKA, Tatsuya TAKIZAWA, Kazuhito SEGAWA, S ...
    2014 Volume 76 Issue 1 Pages 119-122
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 13, 2013
    JOURNAL FREE ACCESS
    A polymorphic tetranucleotide (GAAT)n microsatellite in the first intron of the canine tumor necrosis factor alpha (TNFA) gene was characterized in this study; 139 dogs were analyzed: 22 Beagles, 26 Chihuahuas, 20 Miniature Dachshunds, 24 Miniature Poodles, 22 Pembroke Welsh Corgis and 25 Shiba Inus. We detected the presence of the 4 alleles (GAAT)5, (GAAT)6, (GAAT)7 and (GAAT)8, including 9 of the 10 expected genotypes. The expected heterozygosity (He) and the polymorphic information content (PIC) value of this microsatellite locus varied from 0.389 to 0.749 and from 0.333 to 0.682, respectively, among the 6 breeds. The allelic frequency differed greatly among breeds, but this microsatellite marker was highly polymorphic and could be a useful marker for the canine TNFA gene.
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Laboratory Animal Science
  • Murat BOYACIOGLU, Hulya TURGUT, Cagdas AKGULLU, Ufuk ERYILMAZ, Cavit K ...
    2014 Volume 76 Issue 1 Pages 1-8
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 20, 2013
    JOURNAL FREE ACCESS
    This study was conducted to investigate the prophylactic effects of carnitine against contrast-induced nephropathy (CIN) and its relation to oxidant/antioxidant status in kidney, liver, heart, spleen and lung tissues in a CIN rat model. Twenty-eight adult male Wistar rats were divided into 4 groups, the control, contrast media (CM), carnitine and contrast media+carnitine (CM+carnitine) groups. Animals were placed in individual metabolism cages, and on the 2nd day, rats were deprived of water for 24 hr. On the 3rd day, contrast media were administered to groups CM and CM+carnitine. L-carnitine was administered on days 2, 3 and 4. Histopathological changes were evaluated in the right kidney after euthanization. Superoxide dismutase (SOD) and catalase (CAT) activities and glutathione (GSH) and malondialdehyde (MDA) levels were measured in renal, liver, heart, spleen and lung tissues. The SOD activities in the renal (P<0.05), liver (P<0.001) and spleen (P<0.05) tissues were increased in the carnitine group. The CAT activities in the spleen tissue were decreased (P<0.01) only in the CM group. Renal (P<0.05), liver (P<0.001), spleen (P<0.001) and lung tissue (P<0.01) GSH levels were found to be higher in the carnitine group. In renal, liver and lung tissues, the MDA levels increased in the CM group (P<0.001). The histopathological findings showed that L-carnitine may have a preventative effect in alleviating the negative effects of CIN. Similar to this, L-carnitine may play a major role in the stability of the antioxidant status in the kidney, liver, spleen and lung of the CIN rat model.
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Parasitology
  • Lili CAO, Ronghua CHENG, Lin YAO, Shuxian YUAN, Xinhua YAO
    2014 Volume 76 Issue 1 Pages 9-14
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 20, 2013
    JOURNAL FREE ACCESS
    The Loop-mediated isothermal amplification (LAMP) method amplifies DNA with high simply, specificity, sensitivity and rapidity. In this study, A LAMP assay with 6 primers targeting a highly conserved region of the GRA1 gene was developed to diagnose Toxoplasma gondii. The reaction time of the LAMP assay was shortened to 30 min after optimizing the reaction system. The LAMP assay was found to be highly specific and stable. The detection limit of the LAMP assay was 10 copies, the same as that of the conventional PCR. We used the LAMP assay to develop a real-time fluorogenic protocol to quantitate T. gondii DNA and generated a log-linear regression plot by plotting the time-to-threshold values against genomic equivalent copies. Furthermore, the LAMP assay was applied to detect T. gondii DNA in 423 blood samples and 380 lymph node samples from 10 pig farms, and positive results were obtained for 7.8% and 8.2% of samples, respectively. The results showed that the LAMP method is slightly more sensitive than conventional PCR (6.1% and 7.6%). Positive samples obtained from 6 pig farms. The LAMP assay established in this study resulted in simple, specific, sensitive and rapid detection of T. gondii DNA and is expected to play an important role in clinical detection of T. gondii.
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  • Tatsunori MASATANI, Masahito ASADA, Madoka ICHIKAWA-SEKI, Miho USUI, M ...
    2014 Volume 76 Issue 1 Pages 139-143
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 11, 2013
    JOURNAL FREE ACCESS
    Peroxiredoxins (Prxs) are a family of antioxidant enzymes. Here, we cloned a 2-Cys Prx, BgTPx-1, from the canine Babesia parasite B. gibsoni. Sequence identity between BgTPx-1 and 2-Cys Prx of B. bovis was 81% at the amino acid level. Enzyme activity assay by using recombinant BgTPx-1 (rBgTPx-1) indicated that BgTPx-1 has antioxidant activity. Antiserum from a mouse immunized with rBgTPx-1 reacted with parasite lysates and detect a protein with a monomeric size of 22 kDa and also a 44 kDa protein, which might be an inefficiently reduced dimer. BgTPx-1 was expressed in the cytoplasm of B. gibsoni merozoites. These results suggest that the BgTPx-1 may play a role to control redox balance in the cytoplasm of B. gibsoni.
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  • Phillip Senzo MTSHALI, Ana Mbokeleng TSOTETSI, Matlhahane Molifi Oriel ...
    2014 Volume 76 Issue 1 Pages 145-150
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 20, 2013
    JOURNAL FREE ACCESS
    Babesia bovis and Babesia bigemina are tick-borne hemoparasites causing babesiosis in cattle worldwide. This study was aimed at providing information about the occurrence and geographical distribution of B. bovis and B. bigemina species in cattle from Gauteng province, South Africa. A total of 268 blood samples collected from apparently healthy animals in 14 different peri-urban localities were tested using previously established nested PCR assays for the detection of B. bovis and B. bigemina species-specific genes encoding rhoptry-associated protein 1 (RAP-1) and SpeI-AvaI restriction fragment, respectively. Nested PCR assays revealed that the overall prevalence was 35.5% (95% confidence interval [CI]=± 5.73) and 76.1% (95% CI=± 5.11) for B. bovis and B. bigemina, respectively. PCR results were corroborated by sequencing amplicons of randomly selected samples. The neighbor-joining trees were constructed to study the phylogenetic relationship between B. bovis and B. bigemina sequences of randomly selected isolates. Analysis of phylogram inferred with B. bovis RAP-1 sequences indicated a close relationship between our isolates and GenBank strains. On the other hand, a tree constructed with B. bigemina gp45 sequences revealed a high degree of polymorphism among the B. bigemina isolates investigated in this study. Taken together, the results presented in this work indicate the high incidence of Babesia parasites in cattle from previously uncharacterised peri-urban areas of the Gauteng province. These findings suggest that effective preventative and control measures are essential to curtail the spread of Babesia infections among cattle populations in Gauteng.
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Pathology
  • Sawane MITANI, Akira YABUKI, Mariko SAWA, Hye-Sook CHANG, Osamu YAMATO
    2014 Volume 76 Issue 1 Pages 45-50
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 05, 2013
    JOURNAL FREE ACCESS
    Angiotensin-converting enzyme (ACE) is a key enzyme in the renin-angiotensin system (RAS). ACE2 is a newly identified member of the RAS. The present immunohistochemical study focused on changes in intrarenal ACE and ACE2 immunoreactivity in feline and canine chronic kidney disease (CKD). ACE immunoreactivity was predominantly observed in the brush border of the proximal tubules in dogs and cats. ACE immunoreactivity was lower in CKD kidneys than in normal kidneys, and quantitative analysis demonstrated negative correlations between ACE and renal tissue damage in dogs. ACE2 immunoreactivity was also detected in the proximal tubules; it increased or decreased with CKD in dogs, depending on the renal region assessed. The changes in ACE and ACE2 in CKD were associated with the plasma creatinine concentration in dogs. Findings from dogs with glomerulonephritis were similar to those from dogs with non-glomerulonephritis. The present study suggests that changes in the intrarenal expression of ACE and ACE2 contribute to the pathological mechanisms of canine CKD, but not to the mechanisms of feline CKD.
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  • Miyuu TANAKA, Takeshi IZAWA, Mitsuru KUWAMURA, Miki OZAKI, Tatsuko NAK ...
    2014 Volume 76 Issue 1 Pages 81-84
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 22, 2013
    JOURNAL FREE ACCESS
    Stillbirth and neonatal mortality are significant problems in captive breeding of dolphins, however, the causes of these problems are not fully understood. Here, we report a case of meconium aspiration syndrome (MAS) in a male neonate of bottlenose dolphin (Tursiops truncates) who died immediately after birth. At necropsy, a true knot was found in the umbilical cord. The lungs showed diffuse intraalveolar edema, hyperemic congestion and atelectasis due to meconium aspiration with mild inflammatory cell infiltration. Although the exact cause of MAS in this case was unknown, fetal hypoxia due possibly to the umbilical knot might have been associated with MAS, which is the first report in dolphins. MAS due to perinatal asphyxia should be taken into account as a possible cause of neonatal mortality and stillbirth of dolphin calves.
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  • Hiroyuki OKADA, Kohtaro MIYAZAWA, Shigeo FUKUDA, Yoshifumi IWAMARU, Mo ...
    2014 Volume 76 Issue 1 Pages 103-107
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 27, 2013
    JOURNAL FREE ACCESS
    The aim of this study was to investigate the presence of disease-associated prion protein (PrPSc) in the skeletal muscle of cattle infected with classical bovine spongiform encephalopathy (C-BSE). The study was carried out systematically in 12 different muscle samples from 43 (3 field and 40 experimental) cases of C-BSE; however, muscle spindles were not available in many of these cases. Therefore, analysis became restricted to a total of 31 muscles in 23 cattle. Even after this restriction, low levels of PrPSc were detected in the muscle spindles of the masseter, intercostal, triceps brachii, psoas major, quadriceps femoris and semitendinosus muscles from 3 field and 6 experimental clinical-stage cases. The present data indicate that small amounts of PrPSc are detectable by immunohistochemistry in the skeletal muscles of animals terminally affected with C-BSE.
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  • Ikjae KANG, Hwi Won SEO, Changhoon PARK, Yeonsu OH, Jeehoon LEE, Ok He ...
    2014 Volume 76 Issue 1 Pages 109-112
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 28, 2013
    JOURNAL FREE ACCESS
    The objective of this study was to develop digoxigenin-labeled in situ hybridization (ISH) for the detection of Streptococcus suis in naturally infected pigs with polyserositis and to compare it with biotinylated ISH. Digoxigenin-labeled hybridization signals for S. suis were observed in cells that had infiltrated the fibrous polyserositis and microcolonies in the blood vessels. Mock hybridization showed no hybridization signals for endogenous digoxigenin. Biotinylated hybridization signals for S. suis were observed in cells that had infiltrated the fibrous polyserositis. However, similar hybridization signals were also observed in the fibrous inflammatory area using mock hybridization for endogenous biotin. The present study demonstrated that digoxigenin-labeled ISH is a valuable diagnostic tool for specific detection of S. suis in polyserositic tissues without nonspecific reactions compared with biotinylated ISH.
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  • Maki NISHIMURA, Yumi TOYOTA, Yoshikazu ISHIDA, Hideki NAKAYA, Kyohko K ...
    2014 Volume 76 Issue 1 Pages 123-127
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 09, 2013
    JOURNAL FREE ACCESS
    A 9-month-old steer was autopsied due to recurrent ruminal tympany. A macroscopic examination found an enlarged caudal mediastinal lymph node, and a section of the lymph node revealed necrosis with marked calcification, similar to tuberculous lymphadenitis. Histopathologically, the lesion consisted of multiple coagulative necrotic foci and fibrosis with macrophage, lymphocyte, eosinophil and multinucleated giant cell infiltration. Non-uniform width hyphae were detected in the necrotic area and within the cytoplasm of the multinucleated giant cells, and they were found to be anti-Rhizopus arrhizus antibody positive in an immunohistochemical examination. Therefore, the steer was diagnosed with necrotic caudal mediastinal lymphadenitis due to zygomycetes infection, and inhibition of eructation by the enlarged lymph node was the likely cause of the ruminal tympany.
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  • Kazunori KIMITSUKI, Hassadin BOONSRIROJ, Daisuke KOJIMA, Chun-Ho PARK
    2014 Volume 76 Issue 1 Pages 133-138
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 10, 2013
    JOURNAL FREE ACCESS
    Feline pituitary tumors are rare. An 8-year-old male Japanese domestic cat presented with anorexia and emaciation. The cat died 17 days after admission from progressive neurological symptoms. At necropsy, a pituitary tumor measuring 25 × 18 × 15 mm was found. Microscopically, the tumor was divided into multiple lobules and had grown invasively into the adjacent brain tissue and sphenoid bone. Tumor cells had pleomorphic nuclei with prominent centrally located nucleoli and abundant amphophilic polygonal cytoplasm. Immunohistochemically, the tumor cells stained with anti-adrenocorticotropic hormone (ACTH), α-melanin-stimulating hormone (MSH) and β-endorphin antibodies. Ultrastructurally, the cytoplasm of the tumor cells contained various sized secretory granules. Based on these pathological findings, this tumor was diagnosed as pituitary carcinoma originated from pars intermedia cells.
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Pharmacology
  • Manabu KOIKE, Yasutomo YUTOKU, Aki KOIKE
    2014 Volume 76 Issue 1 Pages 51-56
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 10, 2013
    JOURNAL FREE ACCESS
    Various chemotherapeutic drugs, such as etoposide, and ionizing radiation (IR) have been clinically applied for the treatment of many types of animal and human malignancies. IR and chemotheraputic drugs kill tumor cells mainly by inducing DNA double-strand breaks (DSBs). On the other hand, unrepaired or incorrectly repaired DSBs can lead to chromosomal truncations and translocations, which can contribute to the development of cancer in humans and animals. Thus, it is important to clarify the molecular mechanisms underlying the chemosensitivity or radiosensitivity of mammalian cells in order to develop medical treatments and next-generation chemotherapeutic drugs for cancer. Previously, we established and analyzed cell lines stably expressing chimeric constructs of EGFP and the wild-type Ku80 (XRCC5) protein or its mutant protein to which mutations were introduced by the site-directed mutagenesis. We found that the Ku70 (XRCC6)-binding-site mutations (A453H/V454H) of Ku80 and nuclear localization signal (NLS)-dysfunctional mutations (K565A/K566A/K568A) affected the ability to complement etoposide sensitivity. In this study, we examined the radiosensitivity of these cell lines. We found that either or both amino acid substitutions in two functional domains of Ku80, i.e., Ku70-binding-site mutations (A453H/V454H) and NLS-dysfunctional mutations (K565A/K566A/K568A), affect the ability to complement radiosensitivity. Moreover, these mutations in the two domains of Ku80 affect the DSB-sensing ability of Ku80. These information and Ku80 mutant cell lines used might be useful for the study of not only the dynamics and function of Ku80, but also the molecular mechanism underlying the cellular response to IR and chemotherapeutic drugs in mammalian cells.
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Public Health
  • Tae-Ho CHUNG, Hee-Dong KIM, Young-Sun LEE, Cheol-Yong HWANG
    2014 Volume 76 Issue 1 Pages 73-79
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 20, 2013
    JOURNAL FREE ACCESS
    Helicobacter spp. may have multiple routes of transmission. It is unclear, however, whether the agent is zoonotic and therefore transmitted from an animal reservoir, including dogs. The aim of this population-based study was to assess the relationship between pet ownership or frequent exposure to dogs and Helicobacter spp. infection, especially focusing on HHLO-2 (Helicobacter heilmannii-like organisms type 2) in saliva and feces samples in Korea, using non-invasive genus/species-specific PCR. One hundred twenty-four eligible human subjects and 39 dogs participated in this study. Relativity of contact with dogs and Helicobacter spp. infection diagnosed by genus-specific PCR showed a statistically significant result (P<0.01), but in the relativity analyses between contact with dogs and H. pylori, H. felis and H. bizzozeronii infections diagnosed using species-specific PCR, only Helicobacter felis showed a statistically significant result. Although H. pylori infection showed a statistically significant relativity, no statistically significant association was found between veterinarian subjects and Helicobacter. spp., H. felis and H. bizzozeronii infections. On performing risk factor analyses of HHLO-2 infection by transmission, using matching species, between HHLO-2-positive dog owners and HHLO-2-positive dogs, Helicobacter felis infection showed an extremely significant relativity (P<0.0001), and Helicobacter bizzozeronii may also be a possible significant risk factor (P<0.01). These results suggest that HHLO-2 infection might be a zoonotic infection, because continuous contact with dogs was proved to be correlated with human H. felis and H. bizzozeronii infections in this study.
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  • Yoshimasa SASAKI, Mika HARUNA, Mariko MURAKAMI, Mizuho HAYASHIDA, Nao ...
    2014 Volume 76 Issue 1 Pages 129-132
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 09, 2013
    JOURNAL FREE ACCESS
    This study aimed to confirm that poultry products packed at poultry processing plants have already been contaminated with Listeria monocytogenes and that poultry products contaminated with L. monocytogenes are derived from broiler flocks infected with L. monocytogenes. L. monocytogenes was isolated from 16.8% (58/345) of chicken breast products and 2.3% (8/345) of chicken liver products. In contrast, L. monocytogenes was isolated from the pooled cecal content sample from only 1 (4%) of 25 flocks and was never isolated from any pooled dropping samples collected from 25 farms. The results of our study indicate that cecal content does not seem to be an important source of L. monocytogenes in poultry products.
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Surgery
  • Yasuhiro KURIHARA, Takehito SUZUKI, Motoharu SAKAUE, Ohoshi MURAYAMA, ...
    2014 Volume 76 Issue 1 Pages 15-23
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 27, 2013
    JOURNAL FREE ACCESS
    Adipose tissue-derived stem cells (ADSCs) isolated from adult tissue have pluripotent differentiation and self-renewal capability. The tissue source of ADSCs can be obtained in large quantities and with low risks, thus highlighting the advantages of ADSCs in clinical applications. Valproic acid (VPA) is a widely used antiepileptic drug, which has recently been reported to affect ADSC differentiation in mice and rats; however, few studies have been performed on dogs. We aimed to examine the in vitro effect of VPA on canine ADSCs. Three days of pretreatment with VPA decreased the proliferation of ADSCs in a dose-dependent manner; VPA concentrations of 4 mM and above inhibited the proliferation of ADSCs. In parallel, VPA increased p16 and p21 mRNA expression, suggesting that VPA attenuated the proliferative activity of ADSCs by activating p16 and p21. Furthermore, the effects of VPA on adipogenic, osteogenic or neurogenic differentiation were investigated morphologically. VPA pretreatment markedly promoted neurogenic differentiation, but suppressed the accumulation of lipid droplets and calcium depositions. These modifications of ADSCs by VPA were associated with a particular gene expression profile, viz., an increase in neuronal markers, that is, NSE, TUBB3 and MAP2, a decrease in the adipogenic marker, LPL, but no changes in osteogenic markers, as estimated by reverse transcription-PCR analysis. These results suggested that VPA is a specific inducer of neurogenic differentiation of canine ADSCs and is a useful tool for studying the interaction between chromatin structure and cell fate determination.
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  • Jun TAMURA, Takaharu ITAMI, Tomohito ISHIZUKA, Sho FUKUI, Norihiko OOY ...
    2014 Volume 76 Issue 1 Pages 113-117
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: September 04, 2013
    JOURNAL FREE ACCESS
    Robenacoxib is a newer nonsteroidal anti-inflammatory drug approved for dogs and cats. This study was designed to evaluate the effect of robenacoxib on the minimum alveolar concentration for blunting adrenergic response (MAC-BAR) of sevoflurane in dogs. Sevoflurane MAC-BAR was determined by judging dogs’ response to a noxious electrical stimulus (50 V, 50 Hz and 10 msec) for 10 sec in 6 beagle dogs on two occasions at least a 7-day interval. In each occasion, saline (0.1 ml/kg) or robenacoxib (2 mg/kg) was administered subcutaneously at 1 hr prior to the MAC-BAR determination. Robenacoxib significantly decreased the sevoflurane MAC-BAR (3.44 ± 0.53% for saline vs. 2.84 ± 0.38% for robenacoxib, P=0.039). These results suggest that subcutaneous robenacoxib provides a clinically relevant sparing effect on anesthetic requirement.
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Virology
  • Tatsuya NISHI, Masatoshi OKAMATSU, Kenji SAKURAI, Huy Duc CHU, Long Ph ...
    2014 Volume 76 Issue 1 Pages 85-87
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 27, 2013
    JOURNAL FREE ACCESS
    In August 2012, A/chicken/Vietnam/OIE-2215/2012 (H5N2) was isolated from a chicken in a live bird market (LBM) in Northern Vietnam. Intravenous pathogenicity test revealed that this virus is highly pathogenic in chickens. The PA, HA, NP and M, PB2 and NA, and PB1 and NS genes of the isolate were phylogenetically closely related to those of A/duck/Vietnam/OIE-2202/2012 (H5N1) of clade 2.3.2.1, A/chicken/Vietnam/OIE-1611/2012 (H9N2) and A/chicken/Vietnam/OIE-2468/2012 (H9N2), respectively. All of these viruses were isolated from birds in LBMs in the same province. These results indicate that A/chicken/Vietnam/OIE-2215/2012 (H5N2) is a genetic reassortant and that surveillance of avian influenza in LBMs and stamping out policy are essential for the eradication of highly pathogenic avian influenza viruses from Asia.
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Wildlife Science
  • Hinako SASHIDA, Yoshihisa SUZUKI, Sou ROKUHARA, Kazuya NAGAI, Ryô HARA ...
    2014 Volume 76 Issue 1 Pages 97-101
    Published: 2014
    Released on J-STAGE: February 03, 2014
    Advance online publication: August 26, 2013
    JOURNAL FREE ACCESS
    The prevalence of hemotropic mycoplasmas in wild monkeys is largely unknown. Here, we report the presence of hemoplasmas in blood specimens collected from wild Japanese monkeys (Macaca fuscata) tentatively captured for ecological survey in Mie prefecture, Japan. We examined 9 monkeys using hemoplasma-specific real-time PCR and found all of them positive for a hemoplasma infection. The 16S rRNA gene and 16S to 23S rRNA intergenic spacer region of the hemoplasma detected in wild monkeys were amplified using end-point PCR. The nucleotide sequences of the PCR products were further determined and compared to those of other hemoplasmas. Our examinations revealed a wide prevalence of a hemoplasma strain in Japanese monkeys, which was similar to ‘Candidatus Mycoplasma haemomacaque’ reported in cynomolgus monkeys (Macaca fascicularis). Pathogenic traits of this hemoplasma strain remain unexplored.
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