Journal of Veterinary Medical Science
Online ISSN : 1347-7439
Print ISSN : 0916-7250
ISSN-L : 0916-7250
Volume 80, Issue 8
August
Displaying 1-31 of 31 articles from this issue
Bacteriology
  • Marta DEC, Anna NOWACZEK, Renata URBAN-CHMIEL, Dagmara STĘPIEŃ-PYŚNIAK ...
    2018 Volume 80 Issue 8 Pages 1195-1203
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 06, 2018
    JOURNAL FREE ACCESS
    Supplementary material

    Campylobacteriosis is currently the most frequent zoonosis in humans and the main source of infection is contaminated poultry meat. As chickens are a natural host for Campylobacter species, one strategy to prevent infection in humans is to eliminate these bacteria on poultry farms. A study was conducted to evaluate the probiotic potential of 46 Lactobacillus isolates from chickens faeces or cloacae. All lactobacilli were able to produce active compounds on solid media with antagonistic properties against C. jejuni and C. coli, with L. salivarius and L. reuteri exhibiting particularly strong antagonism. The cell-free culture supernatants had a much weaker inhibitory effect on the growth of Campylobacter, and the neutralization of organic acids caused them to completely lose their inhibitory properties. The ability to produce H2O2 was exhibited by 93% of isolates; most of isolates had a hydrophobic surface, showed excellent survival at pH 2.0 or 1.5, and displayed tolerance to bile; 50% isolates displayed the ability to biofilm formation. Determination of MICs of various antibiotics showed that as much as 80.4% of Lactobacillus isolates were resistant to at least one antimicrobial agent. Seven ultimately selected isolates that met all the basic criteria for probiotics may have potential application in reducing Camylobacter spp. in chickens and thus prevent infections in both birds and humans.

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  • Daneeya EKAPOPPHAN, Athicha SRISUTTHAKARN, Walasinee MOONARMART, Ruang ...
    2018 Volume 80 Issue 8 Pages 1259-1265
    Published: 2018
    Released on J-STAGE: August 22, 2018
    Advance online publication: June 20, 2018
    JOURNAL FREE ACCESS

    This study aims to determine the microbiological profile and risk factors associated with antimicrobial-resistant bacteria in canine severe corneal ulcers. Thirty-two corneal and conjunctival swabs were collected from dogs with diagnosed severe corneal ulcers that presented to Prasu-Arthorn veterinary teaching hospital in Nakhon Pathom, Thailand from June 2015 to June 2016. Microorganisms were identified by means of genotypic and phenotypic approaches. Of 32 ulcers sampled, 26 (81.3%) yielded culturable microorganisms with 24 bacterial isolates and 7 fungal isolates. The most commonly isolated bacteria were Staphylococcus spp. (45.8%, 11/24) and Pseudomonas aeruginosa (20.8%, 5/24). Out of 11 staphylococcal isolates identified, 10 carried the mecA gene providing methicillin resistance. The extended-spectrum β-lactamase (ESBL) encoding genes blaCTX-M and blaVEB-1 were found in an Acinetobacter lwoffii isolate, and blaSHV was found in a P. aeruginosa isolate. Based on the Clinical Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoint criteria, minimum inhibitory concentrations values showed that all bacteria, except for staphylococci, were susceptible to current ophthalmic antibiotics. More than 50% of staphylococci were resistant to all generations of fluoroquinolones and fusidic acid. Chloramphenicol was highly active against staphylococci (81.3% susceptible). The width (P=0.02) and the depth (P=0.04) of ulcers predicted greater risk of yielding resistant bacteria. The identification of antimicrobial-resistant bacteria prompts practitioners to be prudent when choosing ophthalmic antibiotics for severe corneal ulcers.

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  • Javier OJEDA, Miguel SALGADO, Carolina ENCINA, Camila SANTAMARIA, Gust ...
    2018 Volume 80 Issue 8 Pages 1305-1308
    Published: 2018
    Released on J-STAGE: August 30, 2018
    Advance online publication: July 02, 2018
    JOURNAL FREE ACCESS

    A domestic cat dwelling in a dairy cattle farm with haematuria was referred for a physical examination. The examination showed no abnormalities therefore complementary exams were performed. Leukocytosis with neutrophilia, monocytosis and hyperproteinaemia were detected. The urine analysis showed a bacterial infection without ultrasound findings. Serological titers to Leptospira interrogans serovar Pomona and Autumnalis were detected. Molecular analysis demonstrated the presence of Leptospira spp. in urine. The findings were consistent with subclinical leptospirosis. The cattle herd had evidence of Leptospira infection. The microbiological exams confirmed the presence of the Leptospira spp. in urine and serum. According to the evidence presented in this study, cats that dwell within a dairy farm could play a role in the Leptospira infection epidemiologically. The importance of feline leptospirosis must be evaluated with leptospirosis control in livestock.

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Clinical Pathology
  • Kazunori UEDA, Tomoyuki KAWAI, Haruki SENOO, Atsushi SHIMIZU, Akira IS ...
    2018 Volume 80 Issue 8 Pages 1309-1316
    Published: 2018
    Released on J-STAGE: August 30, 2018
    Advance online publication: July 06, 2018
    JOURNAL FREE ACCESS

    Patellar luxation is abnormal displacement of the patella from the femoral trochlear groove. It is seen primarily in small breed dogs and causes pain and limited mobility of the stifle joint. This study aimed to investigate the relationship among patellar luxation, skin extension, and skin collagen fibril diameter. Nine dogs with patellar luxation and five clinically normal dogs were enrolled in the study. We measured the skin extension and investigated the ultrastructure of the skin and patellofemoral ligament by histopathology and transmission electron microscopy. The mean skin extension in dogs with patellar luxation was 18.5 ± 5.5% which is greater than the reference value (14.5%). Mean skin extension in controls was 8.8 ± 1.7% and was within the normal range. In dogs with patellar luxation, histopathology of the skin and patellofemoral ligament showed sparse and unevenly distributed collagen fibers. Transmission electron microscopy identified poorly organized, irregularly shaped, thin collagen fibrils. Collagen fibril thickness in dogs with patellar luxation was significantly less than fibril thickness in controls (P<0.001). There was a significant negative correlation (ρ= −0.863; P<0.001) between skin collagen fibril diameter and skin extension. Skin extension was correlated with patellar luxation and disease severity. Dogs with patellar luxation, joint dysplasia, and hyperextensible skin appear to be pathologically related. This might represent a phenotype of the Ehlers–Danlos syndrome, a hereditary connective tissue disorder in humans.

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Immunology
  • Boondarika NAMBOOPPHA, Kornravee PHOTICHAI, Kanreuthai WONGSAWAN, Phon ...
    2018 Volume 80 Issue 8 Pages 1204-1211
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 06, 2018
    JOURNAL FREE ACCESS
    Supplementary material

    Chicken heterophils generate reactive oxygen species (ROS) molecules to defend against invading pathogens. The present study examined effects of quercetin on chicken heterophils. Heterophils were stimulated with PBS, 50 µM quercetin (QH), PMA or Escherichia coli (EC) and the resulting intracellular ROS molecules were determined. Flow cytometry results showed that cells stimulated with QH, PMA and EC had a higher ROS production. Increases in intracellular ROS molecules were identified in all treatment groups by fluorescence microscopy. Determination of the ability of quercetin to manipulate mRNA expression of ROS subunits was assessed using real-time RT-PCR. Quercetin and other stimulants up-regulated the majority of genes involved in ROS production: CYBB (NOX2), NCF1 (p47phox), NCF2 (p67phox), NOX1 and RAC2. The antioxidant property of QH was explored by measuring mRNA expression of CAT and SOD1. The data indicate increased levels of CAT with all treatments; however, only QH attenuated the expression of the SOD1 gene. To further investigate the effects of ROS-driven inflammation or cell death, IL6, CASP8 and MCL1 genes were preferentially tested. The inflammatory gene (IL6) was profoundly down-regulated in the QH- and PMA-treated groups while EC induced a strikingly high IL6 expression level. Investigation of the known apoptotic (CASP8) and anti-apoptotic (MCL1) genes found down-regulation of CASP8 in the QH- and PMA-treated groups which were contradicted to the MCL1 gene. In conclusion, quercetin can enhance ROS production by regulating the expression of genes involved in ROS production as well as in subsequent processes.

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  • Eliane Macedo SOBRINHO SANTOS, Anna Christina ALMEIDA, Hércules Otacíl ...
    2018 Volume 80 Issue 8 Pages 1317-1324
    Published: 2018
    Released on J-STAGE: August 30, 2018
    Advance online publication: June 25, 2018
    JOURNAL FREE ACCESS

    We conducted an in silico analysis to search for important genes in the pathogenesis of Caseous Lymphadenitis (CL), with prospects for use in formulating effective vaccines against this disease. For this, we performed a survey of proteins expressed by Corynebacterium pseudotuberculosis, using protein sequences collected from the NCBI GenPept database and the keywords “caseous lymphadenitis” and “Corynebacterium pseudotuberculosis” and “goats”. A network was developed using the STRING 10 database, with a confidence score of 0.900. For every gene interaction identified, we summed the interaction score of each gene, generating a combined association score to obtain a single score named weighted number of links (WNL). Genes with the highest WNL were named “leader genes”. Ontological analysis was extracted from the STRING database through Kyoto Encyclopedia of Genes and Genomes (KEGG) database. A search in the GenPept database revealed 2,124 proteins. By using and plotting with STRING 10, we then developed an in silico network model comprised of 1,243 genes/proteins interconnecting through 3,330 interactions. The highest WNL values were identified in the rplB gene, which was named the leader gene. Our ontological analysis shows that this protein acts effectively mainly on Metabolic pathways and Biosynthesis of secondary metabolites. In conclusion, the in silico analyses showed that rplB has good potential for vaccine development. However, functional assays are needed to make sure that this protein can potentially induce both humoral and cellular immune responses against C. pseudotuberculosis in goats.

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  • Ryota KOBA, Sakurako SHOJI, Sawami YOSHIMURA, Yukinobu TOHYA
    2018 Volume 80 Issue 8 Pages 1266-1270
    Published: 2018
    Released on J-STAGE: August 22, 2018
    Advance online publication: July 03, 2018
    JOURNAL FREE ACCESS

    The retinoic acid-inducible gene-I-like receptor (RLR) family is a group of cytosolic RNA helicase proteins that play an important role in sensing viral RNAs. Melanoma differentiation-associated gene 5 (MDA5), an RLR protein, recognizes viral double-stranded RNA and 5’-triphosphate single-stranded RNA in the cytoplasm for the expression of type I interferon (IFN). The expression of MDA5 is also induced by type I IFN. In the present study, we determined the complete coding sequence of the feline MDA5 gene, and analyzed its structure. In addition, we examined tissue expression patterns, inducibilities of the feline MDA5 by polyinosinic-polycytidylic acid and type I IFN, and a functional role of feline MDA5 on type I IFN expression.

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Internal medicine
  • Yasutomo HORI, Kensuke NAKAMURA, Nobuyuki KANNO, Makoto HITOMI, Yohei ...
    2018 Volume 80 Issue 8 Pages 1212-1218
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 22, 2018
    JOURNAL FREE ACCESS

    Alacepril is a relatively novel angiotensin-converting enzyme inhibitor; however, the safety, tolerance, and efficacy of alacepril in terms of cough suppression in dogs with mitral valve disease (MVD) remain unknown. The aim of this study was to investigate the safety, tolerance, and cough suppression efficacy of alacepril in dogs with MVD. This was a multi-center, prospective study. Forty-two dogs with echocardiographic or radiographic evidence of cardiac enlargement in addition to cough were enrolled. Dogs were treated with alacepril (1.0–3.0 mg/kg/day) for at least 4 weeks. One dog (2.4%) developed complications, including appetite loss, lethargy, and vomiting. Thirty-six dogs were re-evaluated after 4 weeks of treatment. Cough resolved or improved in 20 dogs (55.6%) after treatment. Based on the efficacy of alacepril, the dogs were divided into an effective group (n=20) and an ineffective group (n=16). After treatment, the left ventricular end-diastolic internal diameter corrected for body weight was significantly increased from baseline in the ineffective group but was significantly decreased in the effective group. Univariate binomial logistic regression analyses showed that high atrial natriuretic peptide level, N-terminal pro-B-type natriuretic peptide level, and E wave velocity at baseline were significantly correlated with alacepril inefficacy. Alacepril as treatment for MVD is well tolerated in most dogs, and different conditions of cardiac loading may influence the effect of the drug. Alacepril is expected to improve the quality of life of dogs with early stage MVD.

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  • Akihisa SUWA, Tetsuya SHIMODA
    2018 Volume 80 Issue 8 Pages 1271-1276
    Published: 2018
    Released on J-STAGE: August 22, 2018
    Advance online publication: June 25, 2018
    JOURNAL FREE ACCESS

    Hemophagocytic syndrome (HPS) is a clinicopathological entity characterized by histiocytic proliferation, with marked hemophagocytosis in the reticuloendothelial organs. HPS caused by lymphoma is termed lymphoma-associated hemophagocytic syndrome (LAHS), and there are few reports on canine and feline LAHS. The objective of this study was to examine the clinical, diagnostic, and clinicopathologic features of LAHS in six dogs. The diagnostic criteria of LAHS consisted of lymphoma, bicytopenia or pancytopenia in the blood, and increased hemophagocytosis in the reticuloendothelial organs. In one dog, an ocular form of lymphoma was recognized. A splenic form was recognized in two dogs, and a hepatosplenic form was recognized in three dogs. Immunophenotyping revealed T-cell origin in five dogs and B-cell origin in one dog by polymerase chain reaction for antigen receptor rearrangement analysis. Nonspecific esterase stain was performed to differentiate between neoplastic lymphocytes and hemophagocytes. All five dogs with T-cell lymphoma were diagnosed with large granular lymphocyte (LGL) lymphoma. In three cases, palliative therapy with glucocorticoids was conducted, while the other three cases received chemotherapy as well. The survival times for the three dogs with glucocorticoids only were 6, 6, and 10 days and were 30, 54, and 68 days for the three treated with anticancer therapy. The median survival time for the dogs was 20 days. This report indicates that canine LAHS is likely to be caused by LGL lymphoma, and it has an aggressive behavior and poor general prognosis, as seen in humans.

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  • Yui KOBATAKE, Hiroki SAKAI, Hidetaka NISHIDA, Yosuke UEMATSU, Sadatosh ...
    2018 Volume 80 Issue 8 Pages 1277-1280
    Published: 2018
    Released on J-STAGE: August 22, 2018
    Advance online publication: June 21, 2018
    JOURNAL FREE ACCESS

    An 8-year-old castrated male cat presented with acute ataxia and paresis in all four limbs. The cat also exhibited signs of autonomic nervous system impairment. Magnetic resonance imaging revealed swelling of the brachial plexuses bilaterally. Despite treatment, the cat died after 10 days of treatment. A postmortem examination revealed swollen radial nerves and cervical nerve roots in which infiltration of inflammatory cells was histologically confirmed. Additionally, lymphocytic infiltration was found around the blood vessels of the sciatic nerve bundle and the vagus nerve. Histological features were comparable to previously reported brachial plexus hypertrophic neuritis in a cat. Our case was unique in that the autonomic nerves were also involved in addition to the somatic nerves in all four limbs.

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  • Makoto NAKATA, Yasutsugu MIWA, James K. CHAMBERS, Tsubasa SAITO, Kazuy ...
    2018 Volume 80 Issue 8 Pages 1325-1328
    Published: 2018
    Released on J-STAGE: August 30, 2018
    Advance online publication: July 05, 2018
    JOURNAL FREE ACCESS

    A 14-month-old, female mini rex was referred for a detailed examination because of exercise intolerance with associated dyspnea. The thoracic radiograph demonstrated severe cardiac enlargement and elevation of the trachea. The echocardiography revealed dilatations of the right-side heart and pulmonary artery, and the color flow Doppler echocardiography demonstrated an atrial septum defect with left to right shunt, resulting in a disturbed flow. The rabbit died 19 days after the initial presentation, and a necropsy was performed. At the necropsy, a defect, 5 mm in diameter, was detected in the atrial septum. Based on the location of the defect, an ostium secundum type atrial septal defect was diagnosed. This is the first clinical report of atrial septal defect in rabbits.

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Laboratory Animal Science
  • Xiaochuan LIU, Yuwei YANG, Ping JIANG, Xiaohui LI, Yanliang GE, Yang C ...
    2018 Volume 80 Issue 8 Pages 1329-1336
    Published: 2018
    Released on J-STAGE: August 30, 2018
    Advance online publication: May 30, 2018
    JOURNAL FREE ACCESS

    QSOX1 (quiescin-sulfhydryl oxidase 1) is involved in various processes, including apoptosis and the development of breast diseases. Here, we investigated the effect of QSOX1 on the meat quality of Simmental cattle by analyzing the correlation between QSOX1 single nucleotide polymorphisms (SNPs), I2 204 C>T and I2 378 C>T, and certain meat quality traits. The effects of QSOX1 on triglyceride synthesis and cell apoptosis were further validated by gene silencing or overexpression in bovine fetal fibroblasts and mammary epithelial cells. The results showed that I2 204 C>T and I2 378 C>T had significant correlations with loin thickness, hind hoof weight, fat coverage, liver weight, heart weight, marbling and back fat thickness (P<0.05). QSOX1 overexpression also increased triglyceride production and suppressed apoptosis. In summary, QSOX1 is an important factor for meat quality, lipid metabolism, and cell apoptosis, indicating that QSOX1 could be used as a biomarker to assist in breeding cattle with superior meat.

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Pathology
  • Kei IWATA, Kazufumi KASUYA, Kou TAKAYAMA, Yusuke NAKAHARA, Yoshifumi K ...
    2018 Volume 80 Issue 8 Pages 1219-1222
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 11, 2018
    JOURNAL FREE ACCESS

    A 2-year-female arctic fox (Vulpes lagopus) developed anorexia, dehydration, and emaciation during the quarantine period for importation from Norway, and died 17 days later. At necropsy, a fistula was observed on the left gluteal region, and the left eye, left brain, and kidneys were discolored. Histologically, severe diffuse suppurative meningoencephalitis and renal abscesses were detected. Numerous Gram-positive cocci were detected in these lesions. Multidrug-susceptible Staphylococcus pseudintermedius were isolated from the lesions. These results suggest that S. pseudintermedius can cause severe multifocal suppurative meningoencephalitis and nephritis in foxes. This is the first report of multidrug-susceptible S. pseudintermedius meningoencephalitis and nephritis in a fox.

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Physiology
  • Abdelghany HEFNAWY, Mahmoud Atef Youssef HELAL, Ahmed SABEK, Saad SHOU ...
    2018 Volume 80 Issue 8 Pages 1281-1286
    Published: 2018
    Released on J-STAGE: August 22, 2018
    Advance online publication: June 25, 2018
    JOURNAL FREE ACCESS

    The effects of shearing on behavioral patterns, antioxidants, and inflammatory and stress biomarkers was investigated in Ossimi sheep. Clinical parameters and behavioral patterns were recorded, and serum samples were collected pre-shearing and 1, 5, 10 and 15 days post-shearing from 60 Ossimi sheep. The results revealed that grooming and standing idle frequencies were significantly (P<0.01) increased post-shearing. There were significant (P<0.01) increases in IL-6, cortisol, and MDA detected from the 1st to the 10th day post-shearing compared to pre-shearing values. IL-2 and TNF-α significantly (P<0.01) increased from the 1st until the 5th day post-shearing compared to pre-shearing values, while significant (P<0.01) decreases in the values of catalase from the 1st until the 5th day post-shearing compared to pre-shearing values were recorded. Regarding to glutathione reductase, there was a significant (P<0.01) decrease from the 1st until the 10th day post-shearing compared to pre-shearing values. Shearing leads to significant changes in antioxidants, inflammatory, stress biomarkers, and some behaviors in sheep.

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  • Yasunori SHINOZUKA, Kazuhiro KAWAI, Reiichiro SATO, Akito HIGASHITANI, ...
    2018 Volume 80 Issue 8 Pages 1287-1290
    Published: 2018
    Released on J-STAGE: August 22, 2018
    Advance online publication: June 18, 2018
    JOURNAL FREE ACCESS

    The change in milk composition in response to intramammary infusion of Escherichia coli lipopolysaccharide (LPS) was investigated. Four clinically healthy goats were infused with LPS (100 µg) by intramammary administration to the left udder. Clinical manifestations (rectal temperature and physical activity), selected blood parameters (pH and white blood cell count) and milk compositions (somatic cell count and pH) were evaluated at 0 hr (just before challenge) and at multiple time points over the first 24 hr post-challenge. After intramammary LPS challenge, the pH of milk from both udders increased. Thus, this study revealed that LPS-induced mastitis in goat can result in increased pH in milk from the unchallenged (contralateral) udder.

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Public Health
  • Hideya INOUE, Tomoyuki SUZUKI, Masashi HYODO, Masami MIYAKE
    2018 Volume 80 Issue 8 Pages 1223-1227
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 11, 2018
    JOURNAL FREE ACCESS

    In cases of food poisoning, it is important for food sanitation inspectors to determine the causative pathogen as early as possible and take necessary measures to minimize outbreaks. Interviews are usually conducted to obtain epidemiological information to aid in the rapid determination of the cause. However, the current method of determining the causative pathogen has the disadvantage of being reliant upon the experience and knowledge of food sanitation inspectors. Here, we analyzed 529 infectious food poisoning incidents reported in five municipalities in the Kinki region to develop a tool for evaluation using a multinomial logistic regression model, which can predict the causative pathogen based on the patients’ epidemiological information. This tool predicts the most probable cause of the incident by generating a list of pathogens with the highest probability. As a result of leave-one-out cross validation, the agreement ratio with the actual pathogen was 86.4%, and this ratio increased to 97.5% when the agreement was judged by including the true pathogen within the top three pathogens with the highest probability. In cases where the difference of probability between the first and second candidate pathogen was ≥50%, the agreement ratio increased to 94.2%. Using this tool, it is possible to accurately estimate the causative pathogen at an early stage based on patient information, and this will further help narrow the target of investigations to identify causative agent, thereby leading to a prompt identification, which can prevent the spread of food poisoning.

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  • Mioko HONDA, Mamoru SAWAYA, Kensuke TAIRA, Akiko YAMAZAKI, Yoichi KAMA ...
    2018 Volume 80 Issue 8 Pages 1337-1344
    Published: 2018
    Released on J-STAGE: August 30, 2018
    Advance online publication: July 03, 2018
    JOURNAL FREE ACCESS

    Recently, the Sarcocystis parasite in horse and deer meat has been reported to be a causative agent of acute food poisoning, inducing nausea, vomiting and diarrhea. Compared with other causative agents, such as bacteria, viruses and other parasites, in deer meat, the Sarcocystis species parasite, including its stability under various conditions, is poorly understood. In this study, we assessed the viability of Sarcocystis spp. and the activity of their diarrhea toxin (a 15-kDa protein) in deer meat under conditions of freezing, cold storage, pH change and curing. In addition, the heat tolerance was assayed using purified bradyzoites. The results showed that the species lost viability by freezing at −20, −30 and −80°C for <1 hr, heating at 70°C for 1 min, alkaline treatment (pH 10.0) for 4 days and addition of salt at 2.0% for <1 day. Immunoblot assays showed that the diarrhea toxin disappeared together with the loss of viability. However, the parasite survived cooling at 0 and 4°C and acidification (pH 3.0 and 5.0) for more than 7 days with the diarrhea toxin intact. These results provide useful information for developing practical applications for the prevention of food poisoning induced by diarrheal toxin of Sarcocystis spp. in deer meat during cooking and preservation.

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  • Khanh Thuan NGUYEN, Mizuki HASEGAWA, Thu Tam NGUYEN, Thi Minh Tam VO, ...
    2018 Volume 80 Issue 8 Pages 1345-1347
    Published: 2018
    Released on J-STAGE: August 30, 2018
    Advance online publication: June 27, 2018
    JOURNAL FREE ACCESS

    A total of 201 wild geckos from the region of Mekong Delta, Vietnam were collected to determine the viable number and survival period of Salmonella in their feces. Of the 101 samples examined, 24 (23.8%) were Salmonella positive. These 24 geckos excreted Salmonella in their feces in a range of 1 to 8.6 log CFU/g with a mean of 4.5 ± 3.2 log CFU/g. Among the Salmonella serovars, Salmonella Weltevreden was the most predominant serovar (37.5%). Moreover, Salmonella could survive for 6 weeks in gecko feces at room temperature in Vietnam. These results indicate that the wild gecko seems to play an important role as a reservoir for Salmonella and a source of Salmonella infection in humans in Southeast Asian countries.

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  • Mariko MOCHIZUKI, Satoshi NOZAWA, Fumiko MINOWA, Kimihiro OKUBO, Hiroy ...
    2018 Volume 80 Issue 8 Pages 1228-1232
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 13, 2018
    JOURNAL FREE ACCESS

    This study investigates bromine (Br) concentration and its relationship with iodine concentration in serum samples of 86 horses. The mean serum Br concentration in horses pastured on green grass near the seashore was significantly higher (P<0.001) than that in horses pastured in a sand paddock. A significantly negative correlation (r=−0.479, P<0.01) between the serum Br and iodine concentrations was evident in the horses that pastured on green grass. The concentrations of several elements such as sodium and potassium were virtually constant in the serum. In addition, there were elements present below the detection limit of the analytical instruments used. In contrast, it was suggested that geological differences have a marked influence on serum Br concentrations in animals. Thus, we hypothesized that serum Br concentration in horses is a possible indicator reflecting geological differences.

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Theriogenology
  • Xiaohua LIU, Chan LUO, Kai DENG, Zhulian WU, Yingming WEI, Jianrong JI ...
    2018 Volume 80 Issue 8 Pages 1291-1300
    Published: 2018
    Released on J-STAGE: August 22, 2018
    Advance online publication: June 20, 2018
    JOURNAL FREE ACCESS

    The present study was undertaken to examine the effects of cytoplasmic volume on nucleus reprogramming and developmental competence of buffalo handmade cloning (HMC) embryos. We found that both HMC embryos derived from ~150% cytoplasm or ~225% cytoplasm resulted in a higher blastocyst rate and total cell number of blastocyst in comparison with those from ~75% cytoplasm (25.4 ± 2.0, 27.9 ± 1.6% vs. 17.9 ± 3.1%; 150 ± 10, 169 ± 12 vs. 85 ± 6, P<0.05). Meanwhile, the proportions of nuclear envelope breakdown (NEBD) and premature chromosome condensation (PCC) were also increased in the embryos derived from ~150 or ~225% enucleated cytoplasm compared to those from ~75% cytoplasm. Moreover, HMC embryos derived from ~225% cytoplasm showed a decrease of global DNA methylation from the 2-cell to the 4-cell stage in comparison with those of ~75% cytoplasm (P<0.05). Furthermore, the expression of embryonic genome activation (EGA) relative genes (eIF1A and U2AF) in HMC embryos derived from ~225% cytoplasm at the 8-cell stages was also found to be enhanced compared with that of the ~75% cytoplasm. Two of seven recipients were confirmed to be pregnant following transfer of blastocysts derived from ~225% cytoplasm, and one healthy cloned calf was delivered at the end of the gestation period, whereas no recipients were pregnant after the transfer of blastocysts derived from ~75% cytoplasm. These results indicate that the cytoplasmic volume of recipient oocytes affects donor nucleus reprogramming, and then further accounted for the developmental ability of the reconstructed embryos.

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  • Xiao-Xing CHI, Tao ZHANG, Xiao-Li CHU, Jing-Long ZHEN, Dong-Jie ZHANG
    2018 Volume 80 Issue 8 Pages 1348-1355
    Published: 2018
    Released on J-STAGE: August 30, 2018
    Advance online publication: June 22, 2018
    JOURNAL FREE ACCESS

    The effect of genistein on Bcl-2 and Bax protein expression in the ovarian tissue of rats with polycystic ovarian syndrome (PCOS) was evaluated. Sixty rats were divided into six groups. Rats in the Dose group received genistein at a concentration of either 5 (L-gen), 10 (M-Gen) or 20 (H-Gen) mg per kg of body weight per day. The expression of Bcl-2 mRNA and Bax mRNA was determined by in situ hybridization. Bcl-2 and Bax protein concentration was quantified by ELISA. The results showed that the expression of Bcl-2 mRNA and Bcl-2 protein was significantly higher in the high genistein Dose group (H-Gen) when compared to the Model group (MG) (P<0.05). Genistein induced higher expression of the Bcl-2 gene at the transcriptional and translational level. Treatment with genistein resulted in an improvement of ovarian function with Bcl-2 expression being enhanced and Bax expression being suppressed. These alterations may be due to the structural and functional modifications that take place in these cells, and could be related to apoptotic changes that occur in rats with PCOS.

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  • Masanori KOBAYASHI, Tatsuya HORI, Eiichi KAWAKAMI
    2018 Volume 80 Issue 8 Pages 1233-1235
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 22, 2018
    JOURNAL FREE ACCESS

    Two dogs with low plasma testosterone (T) levels and poor semen quality were administered one tablet of 12.5 mg clomiphene citrate orally per day at 2-day intervals for 4 weeks. Plasma T levels, total sperm count, and sperm motility in both dogs temporarily increased between 3 and 6 weeks after the start of treatment. These results indicate that poor semen quality in dogs with low plasma T level can be improved by oral administration of clomiphene citrate.

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Toxicology
  • Ryushin MIZUTA, Yoshio KIKU, Tomohito HAYASHI
    2018 Volume 80 Issue 8 Pages 1301-1304
    Published: 2018
    Released on J-STAGE: August 22, 2018
    Advance online publication: June 25, 2018
    JOURNAL FREE ACCESS

    Cigarette smoke is a strong and independent risk factor for esophageal cancer, while the consumption of cow’s milk has been proposed as a protective factor. The mechanistic role of milk in preventing cancer, however, has not been clarified. We focused our study on acrolein, an abundant unsaturated aldehyde present in cigarette smoke. Acrolein is a highly toxic compound and a putative carcinogen. Using a cell culture system, we found that (1) acrolein caused necrosis in Ramos Burkitt’s lymphoma cells, (2) the necrosis was inhibited by preincubation of acrolein with milk, and (3) acrolein formed adducts with milk proteins. These results indicated the protective effects of cow’s milk against acrolein-induced cytotoxicity via protein-acrolein adduct formation.

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Virology
  • Yahiro MUKAI, Masayuki HORIE, Keizo TOMONAGA
    2018 Volume 80 Issue 8 Pages 1356-1363
    Published: 2018
    Released on J-STAGE: August 30, 2018
    Advance online publication: July 05, 2018
    JOURNAL FREE ACCESS

    Endogenous bornavirus-like elements (EBLs) are sequences derived from bornaviruses (the family Bornaviridae) that are integrated into animal genomes. They are formed through germline insertions of segments of bornaviral transcripts into animal genomes. Because EBLs are molecular fossils of bornaviruses, they serve as precious sources of information to understand the evolutionary history of bornaviruses. Previous studies revealed the presence of many EBLs in bat genomes, especially in vesper bats, and suggested the long-term association between bats and bornaviruses. However, insertion dates of EBLs are largely unknown because of the limitations of available bat genome sequences in the public database. In this study, through a combination of database searches, PCR, and sequencing approaches, we systematically determined the gene orthologies of 13 lineages of EBLs in bats of the genus Myotis and Eptesicus and family Vespertilionidae. Using the above data, we estimated their insertion dates: the EBLs in vesper bats were inserted approximately 14.2 to 53 million years ago. These results suggest that vesper bats have been repeatedly infected by bornaviruses at different points in time during evolution. This study provides novel insights into the evolutionary history of bornaviruses and demonstrates the robustness of combining database searches, PCR, and sequencing approaches to estimate insertion dates of bornaviruses.

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  • Nanako YAMASHITA-KAWANISHI, Ryoma SAWANOBORI, Kosuke MATSUMIYA, Akiko ...
    2018 Volume 80 Issue 8 Pages 1236-1240
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 05, 2018
    JOURNAL FREE ACCESS

    Association of felis catus papillomaviruses (FcaPVs) with feline squamous cell carcinoma (SCC) has been reported worldwide, while there is limited information about FcaPVs in Asia. In this study, 21 feline SCC biopsy samples from cats in Japan were analyzed by PCR with PV consensus primers and type-specific primers for FcaPV type 2 (FcaPV-2), FcaPV-3 and FcaPV-4 designed in this study. Sequence analysis revealed that one sample was FcaPV-3, and two were FcaPV-4. In both FcaPV-4 positive samples, 334th tryptophan in L1 ORF was deleted compared with the reference sequence. Moreover, immunohistochemistry showed that p16 protein was positive in both FcaPV-4 detected samples. This study would contribute to the molecular epidemiological and pathological understanding of FcaPV in Japan.

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  • Tomoha ODAGIRI, Hiroho ISHIDA, Jun-You LI, Maiko ENDO, Tomoya KOBAYASH ...
    2018 Volume 80 Issue 8 Pages 1241-1244
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 05, 2018
    JOURNAL FREE ACCESS

    Influenza (flu) D virus, a possible causative agent of bovine respiratory disease, is genetically classified into three clusters: D/OK-, D/660-, and D/Japan-lineages. To evaluate antigenic heterogeneity among these clusters, we compared antibody titers to each lineage virus using bovine sera collected over time following virus infection. Antibody titers to D/Japan-lineage virus rose rapidly in the acute phase of infection, and were 4 times higher than those to the other clustered viruses. In the later phase of infection, titers to D/Japan-lineage virus were equivalent to those to D/OK-lineage virus, and still higher than those to D/660-lineage virus. These results suggest the existence of common and lineage-specific antigenic epitopes in the hemagglutinin-esterase-fusion protein of flu D viruses.

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  • Manabu NEMOTO, Takashi YAMANAKA, Hiroshi BANNAI, Koji TSUJIMURA, Takan ...
    2018 Volume 80 Issue 8 Pages 1245-1247
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 12, 2018
    JOURNAL FREE ACCESS

    This study compared agar gel immunodiffusion (AGID) protocols for diagnosing equine infectious anemia. Two commercial testing kits were used: one following the Japanese Act on Domestic Animal Infectious Diseases Control and one following the World Organisation for Animal Health (OIE) manual. From 651 samples tested, both protocols gave identical results for 647 samples (23 samples tested positive; 624 tested negative). Non-specific reactions were observed in 21 samples testing negative by the Japanese protocol, but none were observed with the OIE protocol. The kappa coefficient value was 0.962, indicating almost perfect agreement between the two protocols. This study found no difference in diagnostic agreement between the two protocols, but the OIE protocol produced non-specific reactions less frequently than the Japanese protocol.

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Wildlife Science
  • Pavel ŠIROKÝ, Fredric L. FRYE, Nela DVOŘÁKOVÁ, Martin HOSTOVSKÝ, Hynek ...
    2018 Volume 80 Issue 8 Pages 1248-1254
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 11, 2018
    JOURNAL FREE ACCESS

    An adult female of Williams’ mud turtle, Pelusios williamsi long-term captive, that was allegedly caught wild in Kenya was found to have developed papilloma-like skin lesions. Excised tumors were examined histologically after routine processing with hematoxylin and eosin (H & E) stained slides, examined for the presence of viral particles by electron microscopy employing negative staining, and examined for the presence of viral DNA by PCR. Microscopic features in pre-treatment biopsies were fully diagnostic and consistent with multifocal squamous cell papilloma. Viral-type inclusion bodies were not identified. Turtle was found to be infected by reptilian herpesvirus. Association with herpesvirus and vast multiplicity of tumors thwarted surgical solution. An autogenous vaccine was prepared using 5 g of excised fresh tissue, aseptically ground, treated with diluted formalin, centrifuged to obtain a supernatant, and subsequently exposed to UV light. Autogenous vaccine induced substantial areas of necrosis of the papillomatous lesions noted by the loss of cytological architecture, nuclear loss, and by edema. The outer edges of the healing biopsies appeared to be regenerating. Therefore, our vaccine application could be considered as effective. It is difficult to treat and eliminate herpesvirus infection because of its cryptic presence and sudden onset of disease. Successful application of autogenous vaccine could be a potentially promising strategy, which deserves further testing.

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  • Seung-Hun LEE, Dongmi KWAK, Kyoo-Tae KIM
    2018 Volume 80 Issue 8 Pages 1255-1258
    Published: 2018
    Released on J-STAGE: August 10, 2018
    Advance online publication: June 22, 2018
    JOURNAL FREE ACCESS

    This study reports two clinical cases of avian haemosporidian infection caused by a Haemoproteus sp., involving a snowy owl (Bubo scandiacus) and a goshawk (Accipiter gentilis), at a zoo. The snowy owl died after presenting with anorexia, depression and lethargy. A blood smear with Wright’s staining confirmed Haemoproteus infection. Necropsy of the snowy owl revealed hypertrophy of the internal organs, including the liver, gallbladder, kidney and adrenal glands. The goshawk showed anorexia, depression and a lowered head position, and was diagnosed with a Haemoproteus infection based on a blood smear. The goshawk was completely cured by treatment with a combination of atovaquone and proguanil hydrochloride. Both cases showed decreased erythrocytes, hemoglobin and hematocrit values on complete blood count.

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  • Yupadee HENGJAN, Nikom SAE-KOO, Thanmaporn PHICHITRASILP, Yasushige OH ...
    2018 Volume 80 Issue 8 Pages 1364-1367
    Published: 2018
    Released on J-STAGE: August 30, 2018
    Advance online publication: July 02, 2018
    JOURNAL FREE ACCESS

    Flying foxes have been widely studied as they are well-known reservoirs of infectious agents. Understanding their population dynamics might help to explain seasonal patterns of disease prevalence, and contribute towards the conservation of flying fox populations. Therefore, this study explored the annual variation in the number of deaths in P. lylei. The study was conducted from 2015–2017, at a Buddhist temple in Thailand, which is the roosting site of P. lylei. The average total number of bat deaths in a month significantly varied between times of a year. A peak was observed during March and May, which ranged in the period of birthing and lactating. There were no significant differences in the average total number of bat deaths in a month between sexes or age classes across times of a years.

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