Journal of Veterinary Medical Science
Online ISSN : 1347-7439
Print ISSN : 0916-7250
ISSN-L : 0916-7250
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Volume 66, Issue 3
March
Displaying 1-25 of 25 articles from this issue
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Anatomy
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  • Geographical and Functional-Morphological Variations of the Skull in the Gray-Bellied Squirrel
    Hideki ENDO, Junpei KIMURA, Tatsuo OSHIDA, Brian J. STAFFORD, Worawut ...
    2004 Volume 66 Issue 3 Pages 277-282
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.277
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    The geographical variations of the skulls were osteometrically examined in the gray-bellied squirrel (Callosciurus caniceps) from the populations of Korat, Ranong, southernmost Thailand, and Terutau Island. The skull size was larger in northern population than in the southern population in the continental mainland. The zoogeographical influences of the Isthmus of Kra remained unclear, since the plots from Korat population were intermingled with those from southernmost Thailand population in the principal component charts. Although Korat population has been thought to belong to north group, we suggest that Ranong and southernmost Thailand populations may contain individuals from both north and south groups separated by the ancient Kra barrier. Terutau Island population was similar to southernmost Thailand population in skull size, although Terutau population has been isolated in the island and separated from the south group of the Isthmus of Kra. In the proportional analysis the interorbital space was narrower and the binocular sense has been well-developed in Terutau population. It suggests that this population has been highly adapted to arboreal behavior. In contrast, the skull with larger interorbital space was more adaptive for terrestrial life in Korat population. The canonical discriminant analysis could clearly separate the four populations in the scattergrams of discriminant scores.
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Avian Pathology
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  • Histopathological Characteristics of Spindle-cell Proliferative Disease in Broiler Chickens and Its Experimental Reproduction in Specific Pathogen-Free Chickens
    Shigeaki TAKAMI, Masanobu GORYO, Toshiaki MASEGI, Kosuke OKADA
    2004 Volume 66 Issue 3 Pages 231-235
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.231
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    The livers and spleens of 45 broiler chickens (33 to 79 days old) suspected of Marek's disease (MD) at meat inspection were collected and examined histopathologically. Macroscopically, they were enlarged from two to three times, and multiple, small, white areas of plaque or infrequent, large, white nodules were observed in most cases. Only 9 birds (20%) were diagnosed with MD based on the histological examination, while the other 35 birds (78%) had tumor-like proliferative lesions in the Glisson's sheath of the liver and in the white pulp and around the sheathed arteries of the spleen, which differs from the pattern seen in MD. The proliferating cells were mainly spindle-shaped or pleomorphic, and were variable in size with abundant eosinophilic cytoplasm. The disease giving rise to the present lesions was diagnosed tentatively as spindle-cell proliferative disease. Total 50 1-day-old specific pathogen-free chicks by serial passage were inoculated intramuscularly with 0.1 ml of a 10% homogenate of the affected livers or spleens. Microscopically, one inoculated bird, necropsied at 6 weeks of age, had spindle-cell proliferative lesions in the spleen similar to the lesions of naturally occurring spindle-cell proliferative disease. Some birds had tumorous lesions, including renal adenoma, leiomyosarcoma and myxosarcoma. Reverse transcriptase-polymerase chain reaction performed using primers specific for subgroup J avian leukosis virus (ALV) produced specific amplifications of subgroup J ALV genes in 4 of 5 field cases examined.
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Bacteriology
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Biochemistry
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  • Quantitative Changes in Serum Concentration of Bovine Gut Chitinase in Theileria Infection
    Wakako FUJIMOTO, Kazuhiro KIMURA, Masako SUZUKI, Bunei SYUTO, Misao ON ...
    2004 Volume 66 Issue 3 Pages 291-294
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.291
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    Bovine gut chitinase is exclusively produced in the liver and secreted into the blood. In the present study, we established a semi-quantitative method by Western blot analysis for measurement of the chitinase content in blood and examined its alteration during postnatal development and experimental infection with hemoprotozoan parasite in cattle. Its serum levels from 1 week to 1 year of age showed a significant increase only in 3-4-month-old group. The plasma concentration of the gut chitinase was not changed during acute inflammation caused by lipopolysaccharide but increased gradually after a Theileria injection and peaked at 52 days post-infection. It appears that the increase in the blood chitinase levels might be a defensive response in cattle against protozoan infection.
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Clinical Pathology
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  • Prevalence of Autoantibody in Cerebrospinal Fluids from Dogs with Various CNS Diseases
    Naoaki MATSUKI, Kaori FUJIWARA, Satoshi TAMAHARA, Kazuyuki UCHIDA, Sat ...
    2004 Volume 66 Issue 3 Pages 295-297
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.295
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    To examine the prevalence of autoantibody in canine cerebrospinal fluids (CSFs), CSFs were collected from 14 healthy controls and 88 clinical cases with various diseases in the central nervous system (CNS), and were analyzed by an indirect fluorescence antibody test on frozen sections of the cerebrum from normal Beagle dogs. An anti-astrocyte autoantibody was detected in 31 clinical cases with titers ranging from 1:1 to ≥1:100. All tested cases with necrotizing meningoencephalitis (NME: n=22) and granulomatous meningoencephalitis (GME: n=3) possessed the anti-astrocyte autoantibody, while the autoantibody was negative in most cases with other inflammatory CNS diseases. The autoantibody was also detected in 4 of 12 cases with brain tumors. Hence, examination of the autoantibody in the canine CFS would be significant for diagnosing NME and/or GME, as well as for understanding peritumoral events in cases with brain tumors.
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Immunology
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  • A Monoclonal Antibody against Chicken Thrombocytes Reacts with the Cells of Thrombocyte Lineage
    Hiroyuki HORIUCHI, Kazuya TANAKA, Akiko SHIGETA, Kayo YOSHIDA, Kiyoshi ...
    2004 Volume 66 Issue 3 Pages 243-250
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.243
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    A new mouse monoclonal antibody (mAb), HUKT was raised against chicken peripheral blood thrombocytes. The mAb HUKT appeared to detect a specific marker on the surface of chicken thrombocytes. Flow cytometry (FCM) analysis revealed that it did not react with cells from the normal thymus, bursa of Fabricius, six kinds of chicken cell lines, chicken erythrocytes or human platelets. In addition, HUKT+ cells in peripheral blood leukocytes (PBL) were CD45low, Bu-1a- and CD3- cells. Immunoblotting analysis showed that the molecule recognized by HUKT is a monomer with an apparent molecular weight of 150 kDa under non-reducing and reducing conditions. Tissue distribution studies revealed that only cells of thrombocyte lineage in bone marrow and embryonic blood cells were stained by HUKT. The HUKT mAb presented here may be useful for both ontogenetic studies of thrombocyte lineage and immunological studies in the chicken.
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  • Molecular Cloning, Chromosomal Location, and Biological Activity of Porcine Interleukin-21
    Yoshihiro MUNETA, Reiko KIKUMA, Hirohide UENISHI, Tomoaki HOSHINO, Kaz ...
    2004 Volume 66 Issue 3 Pages 269-275
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.269
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    A pig interleukin-21 (IL-21) cDNA was successfully cloned and sequenced from porcine peripheral blood lymphocytes (PBL) stimulated with 10 μg/ml concanavalin A (ConA), 10 μg/ml phytohemagglutinin P (PHA), 50 ng/ml phorbol 12-myristate 13-acetate (PMA), and 0.5 μg/ml anti-porcine CD3 antibody for 48 hr. The open reading frame of the porcine IL-21 cDNA is 459 base pairs in length and encodes 152 amino acids. The predicted amino acid sequence of the porcine IL-21 shows 86.2%, 77.7%, and 58.4% identity to the bovine, human, and murine IL-21, respectively. The porcine IL-21 gene was mapped to porcine chromosome 8 (8q22→q23) by means of fluorescence in situ hybridization and radiation hybrid mapping, where the porcine IL-2 gene had been mapped nearby. The recombinant porcine mature IL-21 expressed by E. coli induced dose-dependent proliferation and IFN-γ production from a human NK cell line, NK0. The porcine IL-21 identified in this study will be helpful for the enhancement of innate immune responses of pigs.
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  • Effects of Antimicrobial Peptides Derived from the Beetle Allomyrina dichotoma Defensin on Mouse Peritoneal Macrophages Stimulated with Lipopolysaccharide
    Maki MOTOBU, Said AMER, Manabu YAMADA, Kikuyasu NAKAMURA, Hisako SAIDO ...
    2004 Volume 66 Issue 3 Pages 319-322
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.319
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    We previously reported that synthetic peptides, RLYLRIGRR-NH2 (peptide A) and RLRLRIGRR-NH2 (peptide B), derived from the beetle Allomyrina dichotoma defensin, showed antimicrobial activity against both Gram-positive and negative bacteria and suppressed lipopolysaccharide (LPS)-induced tumor necrosis factor-α (TNF-α) mRNA expression in a murine macrophage cell line. In this study, inhibitory effects of these peptides in LPS-induced mouse peritoneal macrophage activation were investigated. The supplement of peptide A to macrophages cultured with LPS resulted in a significant decrease in nitric oxide and TNF-α production. Furthermore, NF-κB activation was also blocked by addition of peptide A. These results indicated that peptide A blocked macrophage activation induced by LPS.
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Internal Medicine
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  • A Case of Gastrinoma in a Shih-Tzu Dog
    Ushio FUKUSHIMA, Miyuki SATO, Shozo OKANO, Yoshitaka ARAMAKI, Hiroshi ...
    2004 Volume 66 Issue 3 Pages 311-313
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.311
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    A five-year-old male Shih-Tzu dog presented with severe vomiting and weight loss. The clinical signs were successfully improved by an eight-day treatment with an H2-receptor antagonist, gastrointestinal protectant and antibiotics. Ten days later, however, recurrence of vomiting was seen despite continuous medical treatment. Based on clinical signs and the results of various diagnostic tests including CBC, biochemical analysis, contrast radiography, and endoscopy, a duodenal or pancreatic neoplasm was suspected and exploratory laparotomy was conducted. Some swollen pancreatic regions were found, and biopsy of the pancreas indicated the diagnosis of a gastrin-secreting tumor. Consequently, based on a high serum gastrin level as well as clinical signs and immunohistological findings, we diagnosed the disease as canine gastrinoma, a rare tumor of the pancreas.
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  • Hypercalcemia in a Dog with Resolution of Iatrogenic Cushing's Syndrome
    Momoko NAKAMURA, Yuko KAWAMURA, Michiyo MINEGISHI, Yasuyuki MOMOI, Tos ...
    2004 Volume 66 Issue 3 Pages 329-331
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.329
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    A six-year-old spayed Pug was presented with crust formation and ulcer on the skin. The patient had recieved long-term glucocorticoid therapy for treatment of tentatively diagnosed panniculitis. Severe calcification and pyoderma was observed and the patient was diagnosed with iatrogenic Cushing's syndrome and predonisolone was gradually withdrawn. After the withdrawal, the patient developed marked hypercalcemia (15.3 mg/dl) and finally died from renal failure. It is postulated that the eluted calcium from the calcified lesions may have contributed to the high serum calcium level as the underlying disease was not identified on necropsy.
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  • Expression of LacZ Gene in Canine Muscle by Intramuscular Inoculation of a Plasmid DNA
    Sadatoshi MAEDA, Keitaro OHMORI, Keigo KURATA, Masahiro SAKAGUCHI, Ken ...
    2004 Volume 66 Issue 3 Pages 337-339
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.337
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    DNA immunization induces systemic humoral and cellular immune responses to the antigen encoded by cDNA in a plasmid DNA. In the present study, a plasmid DNA encoding cDNA of β-galactosidase (β-gal), pCAGGS-lacZ, was inoculated intramuscularly to a healthy dog in order to evaluate location and duration of the gene expression. On day 7, the plasmid DNA was found by PCR in the muscle where the plasmid was injected. Furthermore, β-gal expression was detected in the same muscle sample by β-gal staining. However, the plasmid DNA was not detected in any samples collected on days 14, 21 and 28. The present results suggest that duration of the gene expression of β-gal by the plasmid DNA is limited in the muscle in dogs and an efficacy for a gene expression should be evaluated depending on the gene inserted in the plasmid DNA for immunotherapy.
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  • Focal Fatty Liver in a Heifer: Utility of Ultrasonography in Diagnosis
    Tharwat MOHAMED, Shin OIKAWA, Takashi KUROSAWA, Kazushige TAKEHANA, Yo ...
    2004 Volume 66 Issue 3 Pages 341-344
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.341
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    A 30-month-old Holstein heifer presented with a history of decreased appetite and respiratory signs. Sonographic examination of the liver incidentally revealed an area of increased echogenicity between the portal vein and the gallbladder. The lesion was nonspherical and had no mass effect or displacement of the adjacent vessels. Its boundaries, to the liver, were geographic. The liver specimen was histologically compatible with a diagnosis of focal fatty liver change (FFLC). The sonographic features of focal fatty infiltration of the liver are characteristic. Recognition of the ultrasonographic data is important to differentiate FFLC from other lesions.
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Parasitology
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  • Molecular Epidemiological Survey of Benign Theileria Parasites of Cattle in Japan: Detection of a New Type of Major Piroplasm Surface Protein Gene
    Jung-Yeon KIM, Naoaki YOKOYAMA, Sanjay KUMAR, Noboru INOUE, Toshiaki Y ...
    2004 Volume 66 Issue 3 Pages 251-256
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.251
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    Benign Theileria species of cattle are found in most parts of the world. The major piroplasm surface protein (MPSP), a conserved protein in all Theileria species, has been used as a maker for epidemiological and phylogenetical studies of benign Theileria species. Parasites with Ikeda- or Chitose-type MPSP genes are dominant in Japan, but we report here mixed infection cases of Theileria parasites with an additional MPSP type parasite infecting cattle in Abashiri District, Hokkaido. The MPSP gene sequence found in the additional type was closely related to MPSP genes of Theileria parasites found in Southeast Asian countries, including Thailand (Narathiwat) and Indonesia (Java). Theileria parasites from the blood sample were also distinguishable from the Ikeda or Chitose type parasites by the small subunit (SSU) rRNA gene sequence analysis, and they are grouped into the SSU rRNA types C/D found in Korea, North America, and Spain. The present finding of mixed infections of cattle with three different types of Theileria makes epidemiological feature of bovine theileriosis in Japan more complex. We have designed a set of primers specific to this MPSP type in order to conduct further epidemiological study.
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Surgery
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  • Regeneration of Cartilage Tissue by Autologous Chondrocytes Transplantation for Cartilage Defects in a Experimental Bovine Model
    Gonhyung KIM, Masahiro OKUMURA, Darko BOSNAKOVSKI, Taketo ISHIGURO, Ts ...
    2004 Volume 66 Issue 3 Pages 263-267
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.263
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    To evaluate the effects of chondrocytes transplantation on the regeneration of cartilage by intraarticular injection or injection into blood clots at cartilage defects, eight full-thickness cartilage defects were created surgically on the articular surface of each femoral trochlea of two calves. Autologous chondrocytes were isolated individually from the cartilage pieces collected at the creation of defects. And isolated cells were cultured in monolayers for proliferation. Cells were injected into synovial fluid (Group 2, n=11) or into the blood clots at the cartilage defects (Group 3, n=5) of the left femoropatellar joint on weeks 2 and 3, respectively after the operation. The defects (Group 1, n=16) of right femoropatellar joint were left untreated in the control group. After 14 weeks, repaired tissues were evaluated based on gross and histological examinations. In Group 3, more repaired tissues and a better interface between the repaired tissue and host cartilage were observed compared with the results for Groups 1 and 2. Moreover, cartilaginous tissue were observed more in defects of Group 3 than in defects of other groups. In conclusion, the present study suggests that the injection of cells into the blood clot at a cartilage defect might be applicable for the regeneration of damaged cartilage.
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  • Congenital Hepatic Arteriovenous Fistula with Intrahepatic Portosystemic Shunt and Aortic Stenosis in a Dog
    Kazuyoshi KOIDE, Yukiko KOIDE, Yasuhiro WADA, Shigeki NAKANIWA, Yoshih ...
    2004 Volume 66 Issue 3 Pages 299-302
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.299
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    Examination of a 2-month-old male golden retriever presented to the hospital revealed malnutrition, ascites, cardiac murmur and hyperammonemia. Identification of subaortic stenosis and hepatic arteriovenous fistula was made through ultrasonography and angiocardiography. In addition, intrasurgical mesenteric portography showed an intrahepatic portosystemic shunt. The dog did not show portal hypertension and secondary multiple extrahepatic portosystemic shunts. Surgical correction was attempted after medical treatment. The hepatic artery branch which was connected to the hepatic arteriovenous fistula was separated, and completely ligated using silk ligature. However, the separation of the intrahepatic shunt blood vessel was unsuccessful and the dog died 15 hr postoperatively.
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  • Fixation to the Canine Bone of Artificial Implant with New Surface Structure
    Takahiro SENO, Yasuharu IZUMISAWA, Ikuya NISHIMURA, Seiya MAEHARA, Tok ...
    2004 Volume 66 Issue 3 Pages 315-318
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.315
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    Screw and laser (SL) column by making screw threads and forming small holes using laser irradiation on the base metal and conventional beads coating (BC) columns were embedded into the shaft of canine femurs, and compared the implant fixation to the host bone. The interfacial strength in SL columns was almost equivalent as BC columns, and bone-column contact rate was higher than BC columns significantly at twelve weeks after implantation. The newly devised SL surface had almost equivalent bone fixation strength comparable to the conventional BC surface. Also, this surface should provide a useful porous surface for use in artificial joints since there is no risk of surface structure detachment.
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Theriogenology
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  • A Direct Time-Resolved Fluoroimmunoassay (TR-FIA) for Measuring Plasma Estradiol-17β Concentrations in Cattle
    Toru TAKAHASHI, Sanae HAMANAKA, Kei IMAI, Kazuyoshi HASHIZUME
    2004 Volume 66 Issue 3 Pages 225-229
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.225
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    A direct Time-Resolved Fluoroimmunoassay (TR-FIA) system for measuring estradiol-17β (E2) in bovine plasma was developed and evaluated. A 100 μl sample of bovine plasma was used for a TR-FIA without prior extraction and purification. The dose-response curves of reference standards ranged from 0.0625 to 10 pg/well. The minimum detectable concentration of this assay system was 0.625 pg/ml, and 19 pg/ml of E2 caused a 50% reduction of maximum binding. The intra- and inter-assay coefficients of variation were 10.2 and 17.4%, respectively. The plasma E2 concentrations measured by direct TR-FIA correlated closely with those measured after extraction (r=0.939). The results in the present study indicate that the TR-FIA reagent for E2, designed for human research can also be utilized, with some modification, for direct assaying in bovine plasma. This assay type seems to fulfill the requirements for safety, sensitivity, specificity, reproducibility and practical convenience.
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  • Effect of Transfection and Passage Number of Ear Fibroblasts on In Vitro Development of Bovine Transgenic Nuclear Transfer Embryos
    M.M.U. BHUIYAN, Jongki CHO, Goo JANG, Eulsoon PARK, Sungkeun KANG, Bye ...
    2004 Volume 66 Issue 3 Pages 257-261
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.257
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    The objective of this study was to determine if the transfection of human prourokinase (ProU) gene and passage number of transfected ear fibroblasts affected in vitro development of bovine transgenic nuclear transfer (NT) embryos. An expression plasmid for human ProU was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker and human ProU gene into a pcDNA3 plasmid and transfected into bovine ear fibroblasts using a lipid mediated method. Abattoir derived oocytes were enucleated at 18-20 hr post maturation and a single donor cell was transferred into the perivitelline space of a recipient oocyte. After fusion and activation, the couplets were cultured in modified synthetic oviductal fluid (mSOF) medium for 168 hr. In Experiment 1, significantly lower rate in blastocysts formation (10.3%) was observed in transfected donor cells at early passage than that in nontransfected counterparts (22.1%, P<0.05). In Experiment 2, development to blastocysts and GFP expression in blastocysts were not significantly different between early (3-7) and late (8-12) passage donor cells (10.3 vs. 11.3% and 54.5 vs. 41.7%, respectively). This study indicates that in vitro development of bovine transgenic NT embryos is negatively influenced by transfection of human ProU gene into donor fibroblasts. However, passage number of transfected ear fibroblasts does not affect in vitro development of bovine transgenic NT embryos.
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Virology
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  • Development of an ELISA Based on the Baculovirus-Expressed Capsid Protein of Porcine Circovirus Type 2 as Antigen
    Changming LIU, Takeshi IHARA, Tetsuo NUNOYA, Susumu UEDA
    2004 Volume 66 Issue 3 Pages 237-242
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.237
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    The genome of porcine circovirus type 2 (PCV2) contains two major open reading frames, which have been shown to encode the virus capsid and replication-associated proteins. The capsid protein is a major structural protein of the virus; it can be a suitable target antigen for detecting PCV2-specific antibodies to monitor PCV2 infection. To produce the antigen, the capsid protein coding sequence was cloned into a baculovirus transfer vector, and a recombinant capsid (rC) protein of PCV2 was expressed as a combined fusion protein in frame with a C-terminal peptide of six histidines. The affinity-purified rC protein was used as coating antigen to develop an ELISA for detecting the virus-specific antibodies in swine sera. The rC protein-based ELISA (rcELISA) was evaluated by examining a panel of 49 PCV2-positive and 49 PCV2-negative swine sera. In comparative experiments of immunoperoxidase monolayer assay (IPMA) using 102 field sera, there was 89.2% coincidence between data obtained by the rcELISA and IPMA. The rcELISA achieved 88.5% specificity and 89.4% sensitivity for detection of PCV2 antibody in the field sera. The assay showed no cross-reactivity with antibodies to PCV type 1, porcine reproductive and respiratory syndrome virus and porcine parvovirus. The results suggest that the rcELISA is suitable for routine serodiagnosis and epidemiological surveys of PCV2-associated diseases.
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  • High Genetic Stability of TM1 and TM2 Strains of Subtype B Feline Immunodeficiency Virus in Long-Term Infection
    Yasuhiro IKEDA, Takayuki MIYAZAWA, Yorihiro NISHIMURA, Kazuya NAKAMURA ...
    2004 Volume 66 Issue 3 Pages 287-289
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.287
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    To know the genetic changes of feline immunodeficiency virus (FIV) in long-term infection in cats, we inoculated three specific pathogen-free cats with FIV isolates and determined a partial env sequence covering the V3-V5 region. In 2 cats infected with subtype B strains TM1 and TM2, only one amino acid change in region V3 was observed at 9 years post infection (y.p.i.), and no nucleotide substitutions were observed between 9 and 10 y.p.i., indicating that these strains are genetically stable. On the other hand, in a cat infected with subtype A strain Petaluma at 8.7 y.p.i., 3 nucleotide insertions (one amino acid insertion) in region V5, and 1 synonymous nucleotide substitution and 2 non-synonymous nucleotide substitutions in region V5, were observed.
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  • Antigenic Differences between H5N1 Human Influenza Viruses Isolated in 1997 and 2003
    Taisuke HORIMOTO, Nao FUKUDA, Kiyoko IWATSUKI-HORIMOTO, Yi GUAN, Wilin ...
    2004 Volume 66 Issue 3 Pages 303-305
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.303
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    To assess whether the antigenic properties of H5 hemagglutinin (HA) change over time due to antigenic drift, we produced a panel of monoclonal antibodies (mAbs) against the HA of the index H5N1 human influenza A virus, A/Hong Kong/156/97. By immunizing mice with a plasmid expressing this HA and boosting the initial immunization with cell lysates transfected with the plasmid, a total of six hybridomas producing HA-specific mAbs were established: four to the HA1 subunit with hemadsorption-inhibiting activity and two to the HA2 subunit. None of the mAbs to HA1 could bind to the HA of a recent human isolate, A/Hong Kong/213/2003, indicating that there are substantial antigenic differences between the H5N1 human influenza virus isolated in 1997 and that isolated in 2003.
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  • Genetic Variety of Bovine viral diarrhea virus 2 Strains Isolated from Sheep
    Massimo GIANGASPERO, Ryô HARASAWA
    2004 Volume 66 Issue 3 Pages 323-326
    Published: 2004
    Released on J-STAGE: April 21, 2004
    DOIhttps://doi.org/10.1292/jvms.66.323
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    Bovine viral diarrhea virus 2 (BVDV-2) strains, isolated from sheep showing clinical symptoms of border disease, have been evaluated by the palindromic nucleotide substitution (PNS) method at the three variable loci (V1, V2 and V3) in the 5'-untranslated region (UTR) of genomic RNA. The characteristic two base-pairings common to the BVDV-2 species, a C-G pairing which was common to the V1 locus, and a G*U pairing common to the V2 locus, were observed in all tested strains. Strains BD-78 and C413 were identified by a unique C-G pairing at position 4 from the bottom of the V2 stem region, which is characteristic to BVDV-2b. BVDV-2d characteristic U-A pairing at position 18 of the V1 stem region was observed in five strains, Lees, 167 237, 168 149, 173 157 and 175 375. No strains have been assigned to the genotypes BVDV-2a or BVDV-2c. Furthermore, the investigation at the level of the 5'-UTR excluded the application in sheep of the proposed BVDV-2 genetic virulence markers described in cattle. The two specific positions of uracil and cytosine nucleotides related to low or high virulence where indifferently present in the ovine BVDV-2 strains responsible of border disease.
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