Journal of Veterinary Medical Science
Online ISSN : 1347-7439
Print ISSN : 0916-7250
ISSN-L : 0916-7250
Volume 83, Issue 4
April
Displaying 1-28 of 28 articles from this issue
Avian Pathology
  • Takuma ARIIZUMI, Shiro MURATA, Sotaro FUJISAWA, Masayoshi ISEZAKI, Nao ...
    2021 Volume 83 Issue 4 Pages 558-565
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 11, 2021
    JOURNAL OPEN ACCESS
    Supplementary material

    Poultry red mites (PRMs, Dermanyssus gallinae) are harmful ectoparasites that affect farmed chickens and cause serious economic losses in the poultry industry worldwide. Acaricides are used for PRM control; however, some PRMs have developed acaricide-resistant properties, which have indicated the need for different approaches for PRM control. Therefore, it is necessary to elucidate the biological status of PRMs to develop alternative PRM control strategies. Quantitative polymerase chain reaction (qPCR) allows analysis of the biological status at the transcript level. However, reference genes are preferable for accurate comparison of expression level changes given the large variation in the quality of the PRM samples collected in each farm. This study aimed to identify candidate reference genes with stable expression levels in the different blood feeding states and life stages of PRMs. First, we selected candidates based on the following criteria: sufficient expression intensity and no significant expression difference between fed and starved states. We selected and characterized seven candidate reference genes. Among them, we evaluated the gene expression stability between the starved and fed states using RefFinder; moreover, we compared their expression levels in each life-stage and identified two reference genes, Elongation factor 1-alpha (ELF1A)-like and apolipophorins-like. Finally, we evaluated the utility of the candidates as reference genes, and the use of ELF1A-like and apolipophorins-like successfully normalized ATP synthase subunit g -like gene expression. Thus, ELF1A-like and apolipophorins-like could be suitable reference genes in PRMs.

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Clinical Pathology
  • Makoto AKIYOSHI, Masaharu HISASUE, Sakurako NEO, Masami AKIYOSHI
    2021 Volume 83 Issue 4 Pages 689-694
    Published: 2021
    Released on J-STAGE: April 24, 2021
    Advance online publication: March 11, 2021
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    Supplementary material

    This report describes the cases of two Miniature Dachshunds who were suspected to have immune-mediated hemolytic anemia (IMHA) and were treated with immunosuppressive therapy. However, progression of anemia, increases in C-reactive protein (CRP) and total-bilirubin (T-Bil) levels, splenomegaly, transition to nonregenerative anemia, and thrombocytopenia occurred after the treatment. Splenectomy and bone-marrow aspirations were performed subsequently. Both dogs were diagnosed with hemophagocytic syndrome (HPS) associated with IMHA. Unfortunately, they died 9 and 6 days later. These findings indicate that some cases of refractory IMHA have the pathogenicity of HPS. HPS should be included as a differential diagnosis of refractory IMHA concurrent with thrombocytopenia. Continuously elevated CRP and T-Bil levels may be helpful indicators in the detection of HPS associated with IMHA.

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Ethology
  • Simona NORMANDO, Francesca BERTOMORO, Omar BONETTI
    2021 Volume 83 Issue 4 Pages 566-572
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 23, 2021
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    Supplementary material

    Many dogs are relinquished worldwide, so it is important to enhance adoptions’ success. We aimed at investigating factors associated with owners’ satisfaction with adopted dogs, both in general and focusing on galgos. Data on 392 dogs (191 galgos) were gathered using an online survey, investigating dogs’ and owners’ demographics, satisfaction with the adopted dog and post-adoption behavior. Satisfaction was affected by different variables in galgos’ owners as compared to non-sighthound non-podenco dogs’ ones, with only the presence of disobedience on walks negatively affecting satisfaction in both samples. Depending on dogs’ type, the presence of some behavioral problems was associated with decreased satisfaction with the dog (e.g., destructiveness for galgos, or separation problems for non-sighthound non-podenco dogs), whereas that of others increased it (e.g., not being interested in social interactions with dogs for galgos, and shadowing for non-sighthound non-podenco dogs). The variables most often being predictors of the behaviors influencing satisfaction were dog type, with being a galgo as a negative predictor, and dog’s age, with being older as a negative predictor. Further studies on dog adopters’ satisfaction are needed.

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Immunology
  • Kanae NIIMI, Motoko MORIMOTO
    2021 Volume 83 Issue 4 Pages 573-580
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 17, 2021
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    To eliminate pathogens, the initiation of an appropriate immune response is critical. When the gastrointestinal nematode, Heligmosomoides polygyrus (Hp), invades the small intestine, a type-2 cytokine response is initiated; however, this response is not sufficient to clear the infection, and chronic infection can ensue. In this study, the host defense against Hp was investigated in mice with a focus on the role of CD4+ T cells. To this end, tissues from the small intestine and mesenteric lymph node (MLN) were collected every day from just after infection until Day 5 because many previous studies have described the later stages of infection from Day 8 to Day 12, during which Hp returns to the lumen and Th2 cytokine expression reaches its peak. In this study, we focused on investigating the initiation of the type-2 immune response. Our results indicated that the larvae encysted by Day 3. Increased type-2 cytokine gene expression started in the small intestine before Day 2 and increased again on Day 5. Interferon (IFN) γ increased significantly on the second day. Flow cytometry and gene expression analysis of MLN cells revealed that CD4+ T cells were not activated until Day 4. These results suggested that innate immune cells in submucosa are activated immediately after infection, but CD4+ T cells accumulate in the cyst zone later. In addition, IFNγ may have an important role in converting type-2 cytokine-producing cells from innate cells to CD4+ T cells.

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Internal Medicine
  • Hyeryoung JEONG, Hanbin LEE, Jaihyun JUNG, Hyunryung KIM, Jin YU, Hyou ...
    2021 Volume 83 Issue 4 Pages 581-591
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: January 19, 2021
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    Unlike echocardiography, cardiac magnetic resonance imaging (cardiac MRI) results in a near-exact assessment of cardiac structures and function. However, most veterinary studies have focused on dogs with normal cardiac function. We hypothesized that there would be significant differences in cardiac measurements between cardiac MRI and echocardiography when left ventricular (LV) function was abnormal. This study was undertaken to compare measurements of LV function produced by cardiac MRI and echocardiography in dogs whose LV function was altered by pharmacological agents. This study was conducted with six healthy beagle dogs. We increased left ventricular contractility by administration of dobutamine; we decreased cardiac contractility with esmolol. Stroke volume measurements were made by using both cardiac MRI and echocardiography under seven different conditions with general anesthesia: control, three doses of esmolol (100, 200, and 500 µg/kg/min), and three doses of dobutamine (10, 20, and 50 µg/kg/min). Experiments involving each condition were conducted at least 1 week apart. When LV contractility was normal, ejection fraction (EF) and stroke volume (SV), as measured by echocardiography and cardiac MRI, were not significantly different. However, when contractility was changed by pharmacological agents, EF and SV were overestimated by echocardiography, compared to MRI. Evaluation of cardiac function in patients treated with pharmacological agents should be conducted carefully because EF and SV measured by echocardiography can be overestimated, compared with EF and SV obtained by cardiac MRI.

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  • Sung-Yong RHEW, Su-Min PARK, Qiang LI, Ju-Hyun AN, Hyung-Kyu CHAE, Jun ...
    2021 Volume 83 Issue 4 Pages 592-600
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 08, 2021
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    Supplementary material

    Mesenchymal stem cells (MSCs) possess regenerative and immunomodulatory properties and can control the immune dysregulation that leads to β-cell destruction. Stem-cell transplantation could thus manage insulin-dependent diabetes mellitus (IDDM) in dogs. In this pilot study, we aimed to assess canine adipose tissue-derived MSCs (cAT-MSCs) transplantation as a treatment for canine diabetes mellitus. This study included four dogs with over a year of insulin treatment for IDDM, following diagnosis at the Veterinary Medicine Teaching Hospital of Seoul National University. Allogenic cAT-MSCs were infused intravenously three or five times monthly to dogs with IDDM. Blood and urine samples were obtained monthly. General clinical symptoms, including changes in body weight, vitality, appetite, and water intake were assessed. Three of the four owners observed improvement of vitality after stem cell treatment. Two of the four dogs showed improvement in appetite and body weight, polyuria, and polydipsia. C-peptide has increased by about 5–15% in three of the cases, and fructosamine and HbA1c levels have improved in two of the cases. Hyperlipidemia was resolved in two of the dogs, and there was no concurrent bacterial cystitis in any of the dogs. C-peptide secretion and lipid metabolism are associated with diabetic complications. Improvement in these parameters following the treatment suggests that cAT-MSC transplantation in dogs with IDDM might help to improve their insulin secretory capacity and prevent diabetic complications.

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  • Chayanont BHAMARASUTA, Kanyavee PREMRATANACHAI, Nitchanan MONGKOLPINYO ...
    2021 Volume 83 Issue 4 Pages 601-608
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 09, 2021
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    Iron metabolism, hepcidin and some blood profiles were investigated in 13 healthy and 31 chronic kidney disease (CKD) dogs. The study consisted of 2 experiments, experiment I included healthy dogs (CONT) and CKD dogs (stage 2, 3 and 4), while experiment II consisted of anemic CKD dogs subjected to 28-day darbepoetin alfa treatment. The response to darbepoetin alfa could divide anemic CKD dogs into responder (RP) and non-responder (NRP) subgroups. The results from experiment I showed that packed cell volume (PCV) and plasma albumin concentration were significantly lower in CKD dogs of all stages while the total iron binding capacity (TIBC) was lower in only CKD stage 3 and 4 compared with dogs in CONT group. The PCV was related to both TIBC and albumin when considering among all dogs or only in CKD dogs. The hepcidin concentration in CKD dogs with anemia was lower than those without anemia (P<0.05). In experiment II before darbepoetin alfa treatment, RP subgroup had significantly higher iron and TIBC compared with NRP subgroup (P<0.05), the iron concentration was decreased only in RP subgroup after darbepoetin alfa treatment (P<0.05). The percent increase in PCV was correlated with initial TIBC (P<0.01). Plasma hepcidin concentration was not different between CONT and CKD groups and between RP and NRP subgroups both before and after darbepoetin alfa treatment. It is concluded that TIBC and plasma iron concentration play role on anemia and erythropoietic response to darbepoetin alfa treatment in CKD dogs.

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  • Noboru SASAKI, Yoshinori IKENAKA, Yumiko INOUE, Takahiro ICHISE, Noriy ...
    2021 Volume 83 Issue 4 Pages 648-655
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: March 05, 2021
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    Measurement of urinary metanephrines in spot samples is used for the diagnosis of canine pheochromocytoma (PC). We describe a simple analytical method based on liquid chromatography tandem mass spectrometry (LC-MS/MS) for measuring free metanephrine (MN) and normetanephrine (NMN) in spot urine samples. Using the developed method, we evaluated the stability of urinary free-MN and free-NMN at various storing conditions. In addition, we assessed the feasibility of urinary free-MN and -NMN measurement for diagnosing PC. Urine samples were mixed with stable isotope internal standards and thereafter purified by ultrafiltration. The purified samples were analyzed by LC-MS/MS in the multiple reaction monitoring mode after separation on a multimode octa decyl silyl column. The coefficient of variation of free-MN and -NMN measurement was 7.6% and 5.5%, respectively. The linearity range was 0.5–10 µg/l for both analytes. Degradation was less than 10% for both analytes under any of the storage conditions. The median free-NMN ratio to creatinine of 9 PC dogs (595, range 144–47,961) was significantly higher (P<0.05) than that of 13 dogs with hypercortisolism (125, range 52–224) or 15 healthy dogs (85, range 50–117). The developed method is simple and may not require acidification of spot urine. The results of this preliminary retrospective study suggest that the measurement of urinary free metanephrines is a promising tool for diagnosing canine PC.

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  • Luca MALFASSI, Francesca FIDANZIO, Massimo SALA, Silvia MARCARINI, Gio ...
    2021 Volume 83 Issue 4 Pages 695-704
    Published: 2021
    Released on J-STAGE: April 24, 2021
    Advance online publication: September 22, 2020
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    The aims of this pilot study were to evaluate the feasibility and efficacy of high-dose hypofractionated volumetric modulated arc radiotherapy (VMAT) applied to whole pelvic region radiotherapy (WPRT) with multilevel simultaneous integrated boost (MLSIB) combined with piroxicam and chemotherapy for the treatment of canine transitional cell carcinoma (TCC) of the lower urinary tract with muscle invasion TCC. Twelve dogs were enrolled, according to stage, in two groups: group 1, TCC confined to the urinary tract; group 2, TCC with metastasis. The planning target volume dose was tailored from 36 to 42 Gy in 6 fractions. All dogs were prescribed piroxicam and radiosensitizing carboplatin, and six received chemotherapy after radiotherapy. Serial follow-ups with computed tomography and magnetic resonance imaging were performed. Disease control and toxicity effects were evaluated according to the Response Evaluation Criteria in Solid Tumors and Veterinary Radiation Therapy Oncology Group criteria. The treatment was well tolerated, and no high-grade side effects were reported. The median overall survival times for groups 1 and 2 were 1,230 and 150 days, respectively. A considerable percentage of patients in group1 (50%) were still alive at the time of writing this paper, and a longer follow-up could enable a more accurate survival analysis. This preliminary analysis shows that VMAT applied to the WPRT with MLSIB is an effective and safe option for dogs with lower urinary TCC, although the presence of metastases worsens the prognosis.

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  • Mizuki OGAWA, Yasutomo HORI, Nobuyuki KANNO, Naoki IWASA, Takeshi TOYO ...
    2021 Volume 83 Issue 4 Pages 705-715
    Published: 2021
    Released on J-STAGE: April 24, 2021
    Advance online publication: February 05, 2021
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    Plasma N-terminal pro-atrial natriuretic peptide (NT-proANP) concentration increases with progression of myxomatous mitral valve disease (MMVD) in dogs. This multicentre, prospective study compared plasma NT-proANP, N-terminal pro-brain natriuretic peptide (NT-proBNP), ANP, and cardiac troponin I (cTnI) concentrations in dogs with MMVD for their characteristics and discriminatory ability to detect cardiac dilatation and congestive heart failure (CHF). Thirty-six healthy dogs and 69 dogs with MMVD were included. Clinical variables were obtained via physical examination, thoracic radiography, and echocardiography. The discriminatory ability of each cardiac biomarker (CB) to determine the presence or absence of cardiac dilatation (event 1) and CHF (event 2) was evaluated using the receiver operating characteristic curves. Plasma NT-proANP, NT-proBNP, and ANP concentrations showed a significant association with the left atrium/aorta ratio (P<0.01). The area under the curve of plasma NT-proANP and NT-proBNP concentrations were 0.72 and 0.75, respectively in event1 and 0.72 and 0.76, respectively in event2. Plasma NT-proANP and NT-proBNP concentrations showed sensitivity 80.0 and 80.0%; specificity 67.6 and 64.7% in event1 (cutoff value; 8,497.81 pg/ml and 1,453.00 pmol/l, respectively) and sensitivity 85.7 and 81.0%; specificity 60.4 and 64.6% in event2 (cutoff value; 8,684.33 pg/ml and 1,772.00 pmol/l, respectively). In dogs with MMVD, plasma NT-proANP, NT-proBNP, and ANP concentrations increase with left atrial enlargement. Particularly, plasma NT-proANP and NT-proBNP concentrations appeared to be equally useful in the discriminatory ability to detect cardiac dilatation and CHF.

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  • Nao AKIYAMA, Yuka TAGAINO, Ken-ichi WATANABE, Noriyuki HORIUCHI, Yoshi ...
    2021 Volume 83 Issue 4 Pages 609-612
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 09, 2021
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    A 15-day-old Holstein calf with lethargy and tachypnea presented to the Veterinary Teaching Hospital at Obihiro University of Agriculture and Veterinary Medicine for evaluation of suspected congenital heart defect. A Levine grade 6 systolic murmur was noted at right apical site auscultation and phonocardiogram also recorded systolic a murmur. Electrocardiography findings include increased R and S waves, R wave split, and negative T waves without arrhythmia. Echocardiography revealed a single ventricle with a trace of the right ventricular wall, atrioventricular valve regurgitation, and turbulent in a single ventricle. Arterial blood analysis showed a marked decrease in oxygen saturation of 78% and oxygen partial pressure of 44 mmHg. Post-mortem examination confirmed the diagnosis of a single left ventricle.

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  • Tommaso VEZZOSI, Roberto GHINELLI, Paolo FERRARI, Francesco PORCIELLO
    2021 Volume 83 Issue 4 Pages 656-660
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 09, 2021
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    This study reports the echocardiographic reference intervals in the American Staffordshire Terrier (AST). The echocardiographic variables obtained in 57 healthy adult AST were compared with published data from the general canine population and other breeds. In the AST, the left ventricular volumes were lower than values reported in Boxers and Dobermans (P<0.0001), but higher than in small breeds (P<0.0001). The left ventricular ejection fraction was higher than Boxers and Dobermans (P<0.0001), but lower than small breed dogs (P=0.027). The aortic peak velocity values were similar to Boxers (P=0.55) but higher than the general canine population (P<0.0001). The reference intervals presented in this study are clinically useful for an accurate echocardiographic interpretation and screening in the AST.

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  • Takumi KOMIYA, Akihiro MORI, Naohito NISHII, Hitomi ODA, Eri ONOZAWA, ...
    2021 Volume 83 Issue 4 Pages 661-665
    Published: 2021
    Released on J-STAGE: April 09, 2021
    JOURNAL OPEN ACCESS
    Supplementary material

    A 5-year-old castrated male domestic shorthair cat was diagnosed with diabetic ketoacidosis and severe insulin resistance. Although the conventional treatment for diabetic ketoacidosis was provided, the cat required frequent hospitalization because of severe dehydration and repeated diabetic ketoacidosis. We detected anti-insulin antibodies for human in this cat. Serum insulin-binding IgG levels were markedly elevated compared with those in healthy cats and other diabetic cats. We initiated prednisolone to suppress the effects of anti-insulin antibodies. After initiation of prednisolone, the cat was gradually recovered with increasing activity and appetite. Furthermore, satisfactory glycemic control was achieved with combined subcutaneous injection of insulin detemir and insulin degludec.

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Laboratory Animal Science
  • Chungyu PAI, Shunya NAKAYAMA, Yasuyo ITO-FUJISHIRO, Kiichi KANAYAMA, Y ...
    2021 Volume 83 Issue 4 Pages 716-723
    Published: 2021
    Released on J-STAGE: April 24, 2021
    Advance online publication: March 09, 2021
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    Nonhuman primates are commonly used as experimental animals due to their biological resemblance to humans. In patients with cardiac disease, the levels of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) tend to increase in response to cardiac damage, and they are thus used as indicators for the diagnosis of human heart failure. However, no reference values for ANP and BNP have been reported for heart disease in nonhuman primates. In this study, we recorded the age, sex, and body weight of 202 cynomolgus monkeys, and performed evaluations to assess the ANP and BNP levels, electrocardiography and echocardiography, and accordingly divided the monkeys into two groups: healthy monkeys and those with spontaneous cardiac disease. Statistical analysis was performed to determine the relationship of ANP and BNP with the factors of age, sex, and body weight. No significant relationship was found between the levels of ANP and BNP and the factors of age, sex, and body weight. However, both the ANP and BNP levels were significantly different between the healthy monkeys and monkeys with valvular disease. Similar to humans, the ANP and BNP levels tended to increase with the progression of cardiac disease in monkeys. Based on these results, we concluded that ANP and BNP are indicators of cardiac disease in nonhuman primates, and that this nonhuman primate cardiac disease model is applicable for cardiology research in humans.

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  • Sang-A GIM, Dong-Ju PARK, Ju-Bin KANG, Fawad-Ali SHAH, Phil-Ok KOH
    2021 Volume 83 Issue 4 Pages 724-733
    Published: 2021
    Released on J-STAGE: April 24, 2021
    Advance online publication: March 12, 2021
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    Glutamate induces neuronal damage by generating oxidative stress and neurotoxicities. The neurological damage caused by glutamate is more severe during brain development in newborns than in adults. Resveratrol is naturally present in a variety of fruits and medicinal plants and exerts a neuroprotective effect against brain damage. The goal of this study was to evaluate the neuroprotective effects of resveratrol and to identify changed proteins in response to resveratrol treatment during glutamate-induced neonatal cortical damage. Sprague-Dawley rat pups (7 days old) were randomly divided into vehicle, resveratrol, glutamate, and glutamate and resveratrol groups. The animals were intraperitoneally injected with glutamate (10 mg/kg) and/or resveratrol (20 mg/kg) and their brain tissue was collected 4 hr after drug administration. Glutamate exposure caused severe histopathological changes, while resveratrol attenuated this damage. We identified regulated proteins by resveratrol in glutamate-induced cortical damaged tissue using two-dimensional gel electrophoresis and mass spectrometry. Among identified proteins, we focused on eukaryotic initiation factor 4A2, γ-enolase, protein phosphatase 2A subunit B, and isocitrate dehydrogenase. These proteins decreased in the glutamate-treated group, whereas the combination treatment of glutamate and resveratrol attenuated these protein reductions. These proteins are anti-oxidant proteins and anti-apoptotic proteins. These results suggest that glutamate induces brain cortical damage in newborns; resveratrol exerts a neuroprotective effect by controlling expression of various proteins with anti-oxidant and anti-apoptotic functions.

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Parasitology
  • Peru Gopal BISWAS, Yuma OHARI, Uday Kumar MOHANTA, Tadashi ITAGAKI
    2021 Volume 83 Issue 4 Pages 666-670
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 22, 2021
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    We analyzed the nuclear ribosomal internal transcribed spacer (ITS) 1 and ITS2 sequences for Bangladesh isolates of Ascaridia galli, and we determined that the sequences were unreliable as molecular markers for distinguishing A. galli from other Ascaridia species, because the sequences showed high identity with that of A. columbae. However, the ITS1 sequences were available for designing PCR primers distinguishable between Ascaridia galli and Heterakis spp. Bangladesh isolates of A. galli constituted a monophyletic clade along with other geographical isolates in the cytochrome c oxidase subunit I (COI) phylogenetic tree, however, we could not clarify the phylogenetic relationships between A. galli and other Ascaridia spp., because their available sequences in GenBank were very few. The developed PCR method using DNA from A. galli and Heterakis spp. eggs would enable differential diagnosis of the individual infections in the future.

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  • Yen Thi Hoang NGUYEN, Nariaki NONAKA, Haruhiko MARUYAMA, Ayako YOSHIDA
    2021 Volume 83 Issue 4 Pages 671-674
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: March 04, 2021
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    This study aimed to evaluate the sampling method for the detection of Ascaris suum larval DNA in chicken livers using real-time PCR. Chickens were inoculated with A. suum eggs of a single dose (Group A) or repeatedly low doses (Group B). White spots (WSs) were continuously observed on liver from day 3 after the last infection in Group B and day 14 in Group A. In Group A, larval DNA was detected in WS lesions (78.6%) at a significantly higher rate than in the remaining tissue samples (31.3%). In conclusion, applying WS lesions to the assay improved the detection rate of A. suum DNA in chicken livers, especially in the case of a single infection.

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Pathology
  • Yanad ABOU MONSEF, Osman KUTSAL
    2021 Volume 83 Issue 4 Pages 613-621
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 11, 2021
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    Supplementary material

    Hepatic progenitor cells are periportally resident cells capable of differentiating into mature hepatocytes or cholangiocytes to ensure hepatic regeneration. This reaction is termed a ductular reaction. In the present study, regenerative response of the feline liver to different hepatic diseases was investigated immunohistochemically. Regeneration of the liver through hepatocellular replication and proliferation of progenitor cell compartment were comparatively evaluated. Histological and immunohistochemical stainings were conducted on feline liver samples (n=40) representing various hepatobiliary diseases. Cytokeratin (CK) 7, CK19, Proliferating cell nuclear antigen (PCNA), Ki67, and Human hepatocyte marker 1 (Hep Par-1) were used. The presence of progenitor cells within feline livers was proved, both as passive cells in normal liver and as active cells (ductular reaction) in hepatic lesions. CK7 was found to be a suitable antibody for immunohistochemically detecting feline progenitor cells. In acute events, regeneration was predominantly shaped by the division of hepatocytes. In chronic events and severe acute events, hepatocytes lost their ability to divide and regeneration mainly occurred through progenitor cells. Location of the ductular reaction varied between different hepatic diseases. Parenchymal ductular reaction was detected in fulminant hepatitis, chronic hepatitis, hepatocellular lipidosis and metastatic lymphoma, whereas septal ductular reaction was detected in chronic hepatitis and metastatic lymphoma. Ductular reaction exhibited positive staining for Hep Par-1 in chronic and severe acute events. This study indicates the major role played by hepatic progenitor cells in regeneration of the feline liver. Moreover, it shows how the activation pattern of ductular reaction varies according to the hepatobiliary disease type.

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Pharmacology
  • Yasu-Taka AZUMA, Sho SUZUKI, Kazuhiro NISHIYAMA, Taro YAMAGUCHI
    2021 Volume 83 Issue 4 Pages 622-629
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 15, 2021
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    Excessive stress response causes disability in social life. There are many diseases caused by stress, such as gastrointestinal motility disorders, depression, eating disorders, and cardiovascular diseases. Transient receptor potential (TRP) channels underlie non-selective cation currents and are downstream effectors of G protein-coupled receptors. Ca2+ influx is important for smooth muscle contraction, which is responsible for gastrointestinal motility. Little is known about the possible involvement of TRP channels in the gastrointestinal motility disorders due to stress. The purpose of this study was to measure the changes in gastrointestinal motility caused by stress and to elucidate the mechanism of these changes. The stress model used the water immersion restraint stress. Gastrointestinal motility, especially the ileum, was recorded responses to electric field stimulation (EFS) by isometric transducer. EFS-induced contraction was significantly reduced in the ileum of stressed mouse. Even under the conditions treated with atropine, EFS-induced contraction was significantly reduced in the ileum of stressed mouse. In addition, carbachol-induced, neurokinin A-induced, and substance P-induced contractions were all significantly reduced in the ileum of stressed mouse. Furthermore, the expression of TRPC3 was decreased in the ileum of stressed mouse. These results suggest that the gastrointestinal motility disorders due to stress is associated with specific non-selective cation channel.

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Physiology
  • Itsuro KAZAMA
    2021 Volume 83 Issue 4 Pages 675-679
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 22, 2021
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    Hypermagnesemia occurs in elderly people or patients with renal insufficiency after excessive ingestion of magnesium-containing laxatives. In addition to typical electrocardiogram (ECG) findings caused by conduction defects, changes in the ST segments and T waves are also observed in patients with severe hypermagnesemia. This suggested the involvement of similar pathophysiology to acute myocardial infarction, as we previously demonstrated using burn-induced subepicardial injury model in frog hearts. In the present study, by exposing the bullfrog heart to high-magnesium solution, we reproduced prominent ST segment changes in ECG as actually observed in patients with severe hypermagnesemia. In addition to the great increase in the T waves, the ECG showed a marked elevation of the ST segments and the cardiac action potential demonstrated a marked shift of the resting membrane potential to the depolarized side. High-magnesium exposure did not affect the abundance of Na+/K+-ATPase proteins. However, the pharmacological stimulation of Na+/K+-ATPase activity by insulin quickly retrieved the elevated ST segments in ECG. From these results, the functional blockade of Na+/K+-ATPase activity by magnesium ions was thought to be responsible for generating the potassium concentration gradient and the subsequent ST segment changes.

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Public Health
  • Kentaro OKUNO, Sharda Prasad AWASTHI, Germán A. KOPPRIO, Atsushi IGUCH ...
    2021 Volume 83 Issue 4 Pages 630-636
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 22, 2021
    JOURNAL OPEN ACCESS
    Supplementary material

    The aims of this study were to investigate prevalence, O-genotype, and virulence gene profile including Shiga toxin (Stx) 2 gene-subtype of Stx-producing Escherichia coli (STEC) in beef cattle from the Bahía Blanca in Argentina. Rectal swabs were collected from 283 beef cattle in 2012. stx genes were detected in 90 (32%) out of the 283 rectal swabs by stx gene-specific PCR assay. The positive cases were 13 with stx1, 58 with stx2, and 19 with both stx1 and stx2. Among 90 stx gene-positive samples, 45 STEC strains were isolated, which included 3 stx1, 34 stx2, and eight stx1 and stx2 genes positive isolates. O-genotyping grouped 45 STEC strains into 19 different O-genotypes such as Og8, Og145, Og171, Og185 (4 from each), Og22, Og153, Og157 (3 from each) and others. Various stx2 gene-subtypes were identified in 42 STEC strains: 13 positive cases for stx2a, 11 for stx2c, 3 for stx2g, 10 for stx2a and stx2d, 4 for stx2a and stx2c, and 1 for stx2b, stx2c and stx2g. efaI gene, generally prevalent in clinical strains, was detected in relatively high in the STEC strains. These data suggest that stx2a and stx2c were distributed not only in O145 and O157 but also in minor O-genotypes of STEC in Argentina.

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Surgery
  • Hirokazu ISHINO, Iwao SAKONJU
    2021 Volume 83 Issue 4 Pages 637-642
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 05, 2021
    JOURNAL OPEN ACCESS

    We analyzed the mRNA expression of matrix metalloproteinases (MMPs), metalloproteinases with thrombospondin motifs (ADAMTSs), and tissue inhibitors of metalloproteinases (TIMPs) in degenerated and herniated intervertebral disks (IVDs) in chondrodystrophic dogs. In degenerated IVDs, MMP3, 7, 13, and 14, ADAMTS4 and 5, and TIMP1–3 expression were significantly higher vs healthy controls (P<0.05). In herniated IVDs, MMP2, 3, 9, 13, and 14, ADAMTS4 and 5, and TIMP1 expression were significantly greater, and MMP7 expression was significantly lower vs degenerated IVDs (P<0.05). These results suggest that metalloproteinase may play a role in extracellular matrix degradation in IVD degeneration. Decreased MMP7 transcription may prevent proteoglycan degradation and may reduce macrophage infiltration, which might affect the resorption process of herniated IVDs.

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  • Ahram KIM, Naoki SASAKI, Inhyung LEE, Jong-pil SEO
    2021 Volume 83 Issue 4 Pages 643-647
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 11, 2021
    JOURNAL OPEN ACCESS
    Supplementary material

    The purpose of this study was to assess the cardiorespiratory and behavioral responses to the combination of medetomidine and tramadol (M-T) or butorphanol (M-B) in standing laparoscopic ovariectomy in horses. One ovary was removed under M-T and the contralateral ovary was removed under M-B with at least 4 weeks between operations at random. Horses were sedated using intravenous medetomidine (5 µg/kg) followed by tramadol (1 mg/kg) or butorphanol (10 µg/kg) after 5 min. Sedation was maintained through the repeated injection of medetomidine (1 µg/kg) and tramadol (0.4 mg/kg) or medetomidine (1 µg/kg) and butorphanol (4 µg/kg) every 15 min. Cardiorespiratory function and behavioral responses, including, sedation, ataxia, and analgesia, were assessed during the surgery. There were no significant differences in cardiorespiratory values and sedation and analgesia scores between M-T and M-B. Ataxia scores were significantly lower in M-T than in M-B. This result suggests that M-T could maintain smooth and stable standing surgery with minimal cardiorespiratory changes in horses.

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Toxicology
  • Sayaka KITAUCHI, Mizuki MAEDA, Tetsushi HIRANO, Yoshinori IKENAKA, Mis ...
    2021 Volume 83 Issue 4 Pages 746-753
    Published: 2021
    Released on J-STAGE: April 24, 2021
    Advance online publication: February 09, 2021
    JOURNAL OPEN ACCESS
    Supplementary material

    Recently, developmental exposure to clothianidin (CLO) has been shown to cause reproductive toxicity in male mice, but the effects in female mice remain to be clarified. Pregnant C57BL/6N mice were given a no-observed-adverse-effect-level (NOAEL) dose of CLO until weaning. We then examined ovaries of 3- or 10-week-old female offspring. In the CLO-administered group, morphological changes, a decrease in the immunoreactivity of the antioxidant enzyme glutathione peroxidase 4 (GPx4), and activation of genes in the steroid hormone biosynthesis pathway were observed in 3-week-old mice, and decreases of GPx4 immunoreactivity, 17OH-progesterone and corticosterone levels were observed in 10-week-old mice, along with high rates of infanticide and severe neglect, providing new evidence that developmental exposure to CLO affects juvenile and adult mice differently.

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Virology
  • Sittinee KULPRASERTSRI, Keisuke AOSHIMA, Atsushi KOBAYASHI, Takashi KI ...
    2021 Volume 83 Issue 4 Pages 734-741
    Published: 2021
    Released on J-STAGE: April 24, 2021
    Advance online publication: March 15, 2021
    JOURNAL OPEN ACCESS

    Duck Tembusu virus (DTMUV), a neurotropic flavivirus, is a causative agent of severe neurological diseases in different birds. No approved vaccines or antiviral therapeutic treatments are available to date. The poultry industry experiences significant economic losses due to DTMUV infections. Minocycline is a second-generation semi-synthetic tetracycline analogue that is commonly used as an antimicrobial treatment. Experimental studies have indicated the successful protective effects of minocycline against neuronal cell death from neurodegenerative diseases and viral encephalitis. The aim of this study was to investigate the effects of minocycline on DTMUV infection in neurons. Primary duck neurons were treated with minocycline, which exhibited neuroprotective effects via anti-apoptotic function rather than through viral replication inhibition. Minocycline might serve as a potential effective drug in DTMUV infection.

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  • Chiho KANEKO, Yasuyuki KANEKO, Putu Eka SUDARYATMA, Hirohisa MEKATA, Y ...
    2021 Volume 83 Issue 4 Pages 680-684
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 15, 2021
    JOURNAL OPEN ACCESS

    We isolated two pseudorabies virus (PRV) isolates (designated OT-1 and OT-2) from two hunting dogs exhibiting neurological manifestations after eating the flesh of wild boar hunted in Oita prefecture, Kyushu Island, Japan. The isolates corresponded to a previously reported PRV (MY-1 strain) isolated from a hunting dog in neighboring Miyazaki prefecture, and it clustered into genotype II based on the glycoprotein C sequence. Our results suggest that this common PRV strain may have been maintained in wild boars on Kyushu Island even though domestic pigs in this area have attained an Aujeszky’s disease-free status.

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Wildlife Science
  • Chih Chun WU, Makoto NAKATA, James Kenn CHAMBERS, Kazuyuki UCHIDA, Nob ...
    2021 Volume 83 Issue 4 Pages 685-688
    Published: 2021
    Released on J-STAGE: April 09, 2021
    Advance online publication: February 17, 2021
    JOURNAL OPEN ACCESS

    A retrospective study involving eight African pygmy hedgehogs histopathologically diagnosed with granulosa cell tumors was conducted. The age at onset was 2.2–4.5 years, with a median age of 3.6 years. The most common clinical signs were hematuria and abdominal distension, which were observed in >50% cases. Exploratory laparotomy was performed in all cases, and ovariohysterectomy or excision of the abdominal mass was performed. Patients with only hematuria survived for >250 days after surgery, whereas those with initial ascites showed recurrence of ascites or tumor growth and survived for approximately 130 days after surgery. Intraperitoneal injection of carboplatin was performed in three recurrent cases. In one of these three cases, the tumor mass disappeared. Hence, carboplatin can be considered a potential antineoplastic drug for the treatment of granulosa cell tumors.

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  • Hiroko IIMA, Hiroki TERAOKA
    2021 Volume 83 Issue 4 Pages 742-745
    Published: 2021
    Released on J-STAGE: April 24, 2021
    Advance online publication: March 11, 2021
    JOURNAL OPEN ACCESS
    Supplementary material

    An adult Red-crowned crane in captivity that had a displaced compound fracture of the middle upper beak caused by an accident was treated by using Type I-a external skeletal fixator (ESF). The ESF that was equipped with a unilateral epoxy putty fixator and with five half-pins was fixed on the premaxilla bone. The crane with the ESF on the beak was able to feed smoothly by itself. The ESF was removed 49 days after the first surgery. Beak malocclusion, which was observed in the latter half period of fixing, spontaneously improved within one month after removal of the ESF. This is the first successful case of repair of an upper beak fracture in a Red-crowned crane.

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