研究者業績

倉田 修

クラタ オサム  (Osamu Kurata)

基本情報

所属
日本獣医生命科学大学 獣医学部 獣医学科 教授
学位
博士(水産学)

J-GLOBAL ID
200901072207237875
researchmap会員ID
1000192391

論文

 22
  • Goshi Kato, Hayato Nakajima, Kyuma Suzuki, Yuhei Kanzawa, Chihaya Nakayasu, Kosei Taguchi, Osamu Kurata, Motohiko Sano
    Frontiers in immunology 14 1101491-1101491 2023年  
    INTRODUCTION: Temporal elevation of water temperature positively affects immune activity and disease resistance in poikilothermic teleost fish. The ayu, Plecoglossus altivelis, an important fish species for Japanese freshwater fisheries, is usually produced under higher water temperatures than the natural conditions to facilitate rapid growth. However, it has been reported that rearing fish at higher water temperatures inhibits the development of the thymus, suggesting that resistance to infectious diseases is reduced in ayu reared at higher water temperatures. Here, we show that decreased resistance to bacterial cold-water disease and excessive inflammatory responses occurred in ayu reared at 22°C compared with those reared at lower temperatures. METHODS: Ayu larvae were reared at 12°C, 15°C and 22°C for 77 days and fed 3% of their body weight. Thymus index and condition factor was calculated after the fish rearing. Then, ayu reared at the different temperatures were challenged with Flavobacterium psychrophilum and the fish were sampled for histopathology and gene expression analyses. Further, the fish were vaccinated with formalin-killed F. psychrophilum and continuously reared at the three different water temperatures. Serum antibody titer was determined by ELISA and cumulative mortality in each group was recorded after the bacterial challenge. RESULTS: Ayu reared at 22°C showed a significantly lower thymus index and higher condition factor than those reared at lower temperatures. Infiltrated leukocytes and many melanin pigments were frequently observed in the adipose tissues and spleens of ayu reared at 22°C, respectively, but not in those reared at 12°C. The gene expression levels of inflammatory cytokines such as IL-1β, IL-8 and TNFα in the spleen were significantly higher in the 22°C group than in the 12°C group. The cumulative survival rate after challenge with Flavobacterium psychrophilum was 51.7%, 40.0% and 13.3% in the 12°C, 15°C and 22°C groups, respectively. The relative percent survival values of vaccinated fish reared at 15°C and 22°C groups were lower than those reared at 12°C. Moreover, the specific antibody titer of the vaccinated fish was the lowest in the 22°C group and the highest in the 12°C group. DISCUSSION: These results suggest that rearing the fish under high water temperature causes excessive inflammatory responses similar to metabolic inflammation in human obesity, resulting in a decrease of disease resistance. In addition, thymic involution induced by higher water temperature probably leads the poor response to vaccination. The present study provides insights into the physiological and immunological changes of fish under global warming.
  • 倉田 修, 羽地 雅, 川島 拓也, 八木澤 優, 齋藤 哲, 森 竜也
    魚病研究 57(3) 95-102 2022年9月15日  査読有り筆頭著者責任著者
    無機物の食品添加物で構成されるオキシリンクSPのサケ科卵のミズカビ病に対する防除効果を調べた。室内試験においてSaprolegnia属菌5種に対するオキシリンクSPの殺菌効果を検討した結果,30分間の薬浴では10,000倍以下の希釈で殺菌作用を示した。野外試験では,Saprolegnia属菌以外の菌類が発育したサクラマス卵を除き,ニジマスおよびイワナ卵での菌糸発育抑制効果を確認した。魚卵毒性については,生産上大きな被害はないが,ニジマス卵の発眼率および孵化率に影響を与えた。一方,イワナ卵およびサクラマス卵に対する影響は認められなかった。オキシリンクSPによる薬浴はサケ科卵を対象としたミズカビ病の防除に有用であるが,薬浴条件について検討する必要がある。
  • Osamu Kurata, Suyu Miyashita, Shinpei Wada
    Fish Pathology 56(4) 216-219 2021年12月15日  査読有り筆頭著者責任著者
  • Osamu Kurata, Suyu Miyashita, Shusei Toda, Shinpei Wada, Takamitsu Sakai
    Fish Pathology 55(4) 151-161 2021年1月15日  査読有り筆頭著者責任著者
  • Yozo Takada, Kosei Taguchi, Kyuma Suzuki, Akihiko Ashizawa, Koichi Kaji, Shun Watanabe, Shinpei Wada, Osamu Kurata
    Fish Pathology 55(2) 31-37 2020年6月  査読有り責任著者

MISC

 75
  • Takeshi Komine, Saralee Srivorakul, Mitsumi Yoshida, Yoshiaki Tanaka, Yuma Sugimoto, Mari Inohana, Hanako Fukano, Yoshihiko Hoshino, Osamu Kurata, Shinpei Wada
    Journal of fish diseases 46(5) 507-516 2023年5月  
    Mycobacterium marinum is a slow-growing, photochromogenic nontuberculous mycobacterium, which can cause mycobacteriosis in various animals, including humans. Several cases of fish mycobacteriosis have been reported to date. Mycobacterium marinum has also been isolated from aquatic environmental sources such as water, sand, biofilms, and plants in the natural environments. Hence, we hypothesized that a wide variety of sources could be involved in the transmission of M. marinum. In this study, we tested this hypothesis by isolating M. marinum from various sources such as fish, invertebrates, seagrass, periphytons, biofilms, sand, and/or water in two aquaria in Japan and conducting a phylogenetic analysis based on single-nucleotide polymorphisms (SNPs) using whole-genome sequences of the isolated strains. The analysis revealed that the strains from animal and environmental sources belonged to the same clusters. This molecular-based study epidemiologically confirmed that various sources, including fish, invertebrates, and environmental sources, could be involved in transmission of M. marinum in a closed-rearing environment. This is the first report where M. marinum was isolated from different sources, and various transmission routes were confirmed in actual cases, which provided essential information to improve the epidemiology of M. marinum.
  • Takeshi Komine, Yuko Matsuoka, Mari Inohana, Osamu Kurata, Shinpei Wada
    Microbiology resource announcements 12(3) e0113522 2023年3月16日  
    Mycobacterium chelonae is a nontuberculous mycobacterium that causes infections in various animals, including humans. In this study, we report the draft genome sequence of M. chelonae subsp. bovis strain NJB1701, which was isolated from a Baikal seal (Pusa sibirica) in captivity in Japan.
  • Takeshi Komine, Hyogo Ihara, Mari Inohana, Jennifer Caroline Kwok, Akane Shimizu, Tsumugi Terasawa, Ayaka Miyazaki, Saralee Srivorakul, Hajime Iwao, Sachiko Harada, Mitsunori Yoshida, Yoshihiko Hoshino, Osamu Kurata, Hanako Fukano, Shinpei Wada
    Frontiers in veterinary science 10 1248288-1248288 2023年  
    INTRODUCTION: Mycobacterium montefiorense is one of the causes of non-tuberculous mycobacterial infections in moray eels and salamanders. Although M. montefiorense infection could be a threat to salamanders, little information is available regarding this pathogen and associated infection. This study aimed to provide fundamental information regarding M. montefiorense and its infection in salamanders. METHODS: Nine M. montefiorense strains isolated from three species of salamanders, namely, Japanese black salamander (Hynobius nigrescens), Hakuba salamander (H. hidamontanus), and Tohoku hynobiid salamander (H. lichenatus), between 2010 and 2018, were characterized based on phenotypic and genetic examination. We also pathologically observed salamanders infected with the M. montefiorense strains, including Hakuba salamanders and Tohoku hynobiid salamanders. RESULTS: The microbiological and chemical characteristics of the M. montefiorense salamander and an eel strain (reference strain) matched. Susceptibility testing for antimicrobials suggested that clarithromycin may be effective. Regarding disinfectants, phtharal, peracetic acid, glutaral, sodium hypochlorite, and benzalkonium chloride may be effective. Phylogenetic analyses revealed that the strains isolated from salamanders in 2014 and 2018 were genetically closely related, which could indicate an outbreak. The main gross findings in infected salamanders include skin ulcerative lesions or nodules in the enlarged liver. Microscopically, multifocal to coalescent granulomatous lesions composed of massive macrophages containing numerous acid-fast bacilli were prominently observed in the liver. CONCLUSION: This study contributes to our understanding of the genetic diversity and phenotypic characteristics of M. montefiorense, as well as the pathology of the infection.
  • Mari Inohana, Takeshi Komine, Yoshiaki Tanaka, Osamu Kurata, Shinpei Wada
    Journal of fish diseases 46(1) 47-59 2023年1月  
    Mycobacterium marinum is a prevalent nontuberculous mycobacterium (NTM)-infecting teleosts. Conversely, little is known about mycobacteriosis in elasmobranchs, and M. marinum infection has never been reported from the subclass. This study investigated the histopathological characteristics and localization of this mycobacterium through molecular analysis of two captive sharks, a scalloped hammerhead Sphyrna lewini and a Japanese bullhead shark Heterodontus japonicus, exhibited in the same aquarium tank. We detected genital mycobacteriosis caused by M. marinum infection using molecular analyses, including polymerase chain reaction (PCR) and DNA sequencing targeting the 60 kDa heat-shock protein gene (hsp65), and peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) targeting the 16S rRNA gene. Both sharks showed granulomas in connective tissues of the gonads without central necrosis or surrounding fibrous capsules, which is unlike the typical mycobacterial granulomas seen in teleosts. This study reveals that elasmobranchs can be aquatic hosts of M. marinum. Because M. marinum is a representative waterborne NTM and a potential zoonotic agent, cautious and intensive research is needed to overcome a lack of data on the relationship between NTM and the aquatic environment in association with this subclass of Chondrichthyes.
  • Takeshi Komine, Hyogo Ihara, Kentaro Ono, Mitsumi Yoshida, Yuma Sugimoto, Mari Inohana, Hanako Fukano, Osamu Kurata, Shinpei Wada
    The Journal of veterinary medical science 84(12) 1617-1620 2022年11月18日  
    In 2019, several aquarium-reared fish died at a sea life park in Japan. Necropsy revealed micronodules on the spleen in the dotted gizzard shad (Konosirus punctatus). Seven of 16 fish exhibited microscopic multifocal granulomas associated with acid-fast bacilli in the spleen, kidney, liver, alimentary tract, mesentery, gills, and/or heart. Bacterial cultures yielded isolates from the dotted gizzard shad and a Japanese sardine (Sardinops melanostictus). Microbiological and molecular biological examinations revealed the isolates as Mycobacterium pseudoshottsii. To our knowledge, this is the first isolation of M. pseudoshottsii from aquarium-reared fish.
  • Takeshi Komine, Hyogo Ihara, Hanako Fukano, Yoshihiko Hoshino, Osamu Kurata, Shinpei Wada
    Microbiology resource announcements 11(11) e0070222 2022年11月17日  
    Mycobacterium montefiorense is a nontuberculous mycobacterium that causes infections in fish and salamanders. Here, we report annotated draft genome sequences of eight strains that were isolated in 2014 and 2018 from salamanders reared in an aquarium in Japan.
  • Takeshi Komine, Hanako Fukano, Mitsunori Yoshida, Mari Inohana, Yoshihiko Hoshino, Osamu Kurata, Shinpei Wada
    Microbiology resource announcements 11(12) e0078522 2022年11月9日  
    Mycobacterium pseudoshottsii, a slow-growing nontuberculous mycobacterium, has been isolated from wild and cultured fish. We report here the complete genome and partial megaplasmid sequences of a strain isolated from an aquarium-reared Japanese sardine (Sardinops melanostictus) in Japan, M. pseudoshottsii NJB1907-Z4.
  • Ushine, N, O. Kurata, Y. Tanaka, S. M, M. Nakayama, M. Ishizuka, T. Kato, S.-I. Hayama
    Avian Conservation and Ecology 17(2) 40 2022年11月  査読有り
  • Takeshi Komine, Hanako Fukano, Mari Inohana, Yoshihiko Hoshino, Osamu Kurata, Shinpei Wada
    Microbiology resource announcements 11(10) e0085122 2022年10月20日  
    Mycobacterium marinum is a ubiquitous nontuberculous mycobacterium that causes infections in various animals. Here, we report the annotated draft genome sequences of 25 strains isolated from vertebrates, invertebrates, and environmental components in aquaria and an aquaculture farm in Japan, sampled between 2015 and 2020.
  • Komine Takeshi, Srivorakul Saralee, Tomaru Kentaro, Tanaka Yoshiaki, Inohana Mari, Fukano Hanako, Kurata Osamu, Wada Shinpei
    Fish Pathology 56(4) 187-198 2021年12月15日  査読有り
  • Nana Ushine, Osamu Kurata, Yoshikazu Tanaka, Tatsuo Sato, Yoshihiro Kurahashi, Shin-Ichi Hayama
    The Journal of veterinary medical science 82(11) 1619-1626 2020年9月21日  査読有り
    In order to elucidate the relationship between migration period and immunity related to susceptibility, we conducted research on Black-headed gulls (Chroicocephalus ridibundus). We captured 260 gulls and collected their peripheral blood. Their leukocyte (WBC) count, percentages of heterophils (Het) and lymphocytes (Lym), heterophil and lymphocyte ratio (H/L ratio), and CD4 and CD8α expression levels (CD4 and CD8α, respectively) were quantitatively analyzed over three migration periods (Autumn migration, Wintering, Spring migration). In Adult gulls, WBC counts and CD4 levels significantly increased. Moreover, the Het and H/L ratio decreased from the Autumn migration to Wintering. Conversely, only WBC counts and CD4 levels measurements significantly decreased from Wintering to Spring migration (P<0.05). The tested parameters of the Tokyo-bay population show a greater significant difference than the measurements of immunity of the Mikawa-bay population. This study suggests that the migratory period has a negative effect on an aspect of the immune system. Including the period-difference in the immune systems in the local population, it is necessary to investigate the relationship between the ecology of migratory birds and their immunity.
  • Hanako Fukano, Mitsunori Yoshida, Akane Shimizu, Hajime Iwao, Yukie Katayama, Tsutomu Omatsu, Tetsuya Mizutani, Osamu Kurata, Shinpei Wada, Yoshihiko Hoshino
    Genome announcements 6(21) 2018年5月24日  査読有り
    Mycobacterium montefiorense is a member of the Mycobacterium simiae complex, the largest group of nontuberculous mycobacteria. Here, we report the genome sequence of M. montefiorense isolate BS, isolated from diseased Japanese black salamander (Hynobius nigrescens) reared in an aquarium in Japan. This is the first reported case of an M. montefiorense infection in an amphibian.
  • Lau LM, Hatai K, Fukano H, Kurata O
    Journal of aquatic animal health 30(4) 239-244 2018年4月  査読有り
    © American Fisheries Society 2018. In September 2014, a freshwater oomycete was first isolated from Asian Seabass Lates calcarifer fry that were reared in freshwater at a fish hatchery in Sabah, Malaysia. A fungal strain was isolated from infected fry by using glucose yeast extract (GY) agar. From morphological identification, the strain belonged to the genus Achlya based on the mode of zoospore release. Molecular phylogenetic analysis of the internal transcribed spacer region sequences from the strain showed high similarity (99-100%) to Achlya oblongata. The isolate was able to grow on GY agar incubated at 15-35°C, in GY broth adjusted to pH 3.0-11.0, and in up to 1.0% NaCl. This is the first report of Achlya infection in freshwater-reared Asian Seabass in Malaysia.
  • 倉田 修
    魚病研究 = Fish pathology 52(4) 168-171 2017年12月  
  • Hanako Fukano, Shinpei Wada, Osamu Kurata, Kinya Katayama, Nagatoshi Fujiwara, Yoshihiko Hoshino
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 67(8) 2811-2817 2017年8月  査読有り
    A previously undescribed rapidly growing, non-pigmented mycobacterium was identified based on biochemical and nucleic acid analyses, as well as growth characteristics. Seven isolates were cultured from samples collected from five thread-sail filefish (Stephanolepis cirrhifer) and two farmed black scraper (Thamnaconus modestus). Bacterial growth occurred at 1535 degrees C on Middlebrook 7H11 agar. The bacteria were positive for catalase activity at 68 degrees C and urease activity, intermediate for iron uptake, and negative for Tween 80 hydrolysis, nitrate reduction, semi-quantitative catalase activity and arylsulfatase activity at day 3. No growth was observed on Middlebrook 7H11 agar supplemented with picric acid, and very little growth was observed in the presence of 5% NaCl. a-and a-mycolates were identified in the cell walls, and a unique profile of the fatty acid methyl esters and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) profiles of the protein and cell-wall lipids were acquired. Sequence analysis revealed that the seven isolates shared identical sequences for the 16S rRNA, rpoB, hsp65, recA and sodA genes. Phylogenetic analysis of the five gene sequences confirmed that the isolates were unique, but closely related to Mycobacterium chelonae. Antibiotic susceptibility testing revealed the minimum inhibitory concentration (MIC) of clarithromycin against this novel species was &lt;0.25 mu g ml(-1), which was lower than that for Mycobacterium salmoniphilum. The hsp65 PCR restriction enzyme analysis pattern differed from those of M. chelonae and M. salmoniphilum. Based on these findings, the name Mycobacterium stephanolepidis sp. nov. is proposed for this novel species, with the type strain being NJB0901(T) (= JCM 31611(T) = KCTC 39843(T)).
  • Hanako Fukano, Mitsunori Yoshida, Yukie Katayama, Tsutomu Omatsu, Tetsuya Mizutani, Osamu Kurata, Shinpei Wada, Yoshihiko Hoshino
    Genome Announcements 5(33) 2017年8月1日  査読有り
    © 2017 Fukano et al. Mycobacterium stephanolepidis is a rapid-growing nonpigmented species isolated from marine teleost fish (Stephanolepis cirrhifer) and is closely related to Mycobacterium chelonae. Here, we report the complete sequence of its genome, comprising a 4.9-Mb chromosome. The sequence represents essential data for future phylogenetic and comparative genome studies of this fish pathogen.
  • Yih Nin Lee, Kishio Hatai, Osamu Kurata
    Fish Pathology 52(1) 31-37 2017年  査読有り
    Mass mortality of eggs and larvae of the mud crab Scylla tranquebarica occurred at a hatchery in Sabah, Malaysia in June 2014. The mortality of the larvae reached nearly 100% within 5 days post-hatching. We isolated a pathogenic Peronosporomycetes (Oomycota) from the mud crab eggs and larvae using peptone-yeast-glucose-seawater (PYGS) agar, and then randomly selected the strain IPMB 1402 from the isolates for further study. We observed the formation of fragments inside the hyphae, a characteristic of Haliphthoros species. The strain morphologically most resembled Haliphthoros milfordensis, but comparisons of DNA sequences of the ITS1 region confirmed that the strain was not identical to that species. We classified the strain into a clade with Haliphthoros sp. group 2 (comprising strains NJM 0440, NJM 0449 and NJM 0535) after determining 97-100% intraspecific similarity. Based on distinctive morphological and molecular characteristics, the strain IPMB 1402 is here assigned to the genus Haliphthoros and designated Haliphthoros sabahensis sp. nov. Like other recognized species of Haliphthoros, it is an obligate marine fungus growth was observed in pH 4-9, and optimum growth temperature was 25-30°C. The strain was experimentally pathogenic to nauplii of the brine shrimp Artemia salina.
  • Y. N. Lee, K. Hatai, O. Kurata
    BULLETIN OF THE EUROPEAN ASSOCIATION OF FISH PATHOLOGISTS 37(6) 226-234 2017年  査読有り
    Fungal infection occurred during mud crab spawning in a hatchery and caused almost 100% mortality in mud crab larvae after 5 days post-hatching. A fungus was isolated from eggs and larvae using PYGS agar and named IPMB 1603. During morphological identification, the strain IPMB 1603 was observed to produce a fragment inside the hyphae and the zoospores swam away from the tips of the discharge tube on several occasions, this closely resembling the genus Haliphthoros. Following comparisons made on the nucleotide sequence of the ITS1 region, the strain IPMB 1603 was identified belonging to a cluster of Haliphthoros milfordensis sharing 97-100% similarity. In this study, the strain IPMB 1603 was found to be an euryhaline fungus as the strain was able to grow on PYG agar containing NaCl. Strain IPMB 1603 only grew at certain salinity ranges, in which on PYGS agar contained seawater higher than 10 ppt. The optimum temperature range for fungal growth of strain IPMB 1603 was 30-35 degrees C. Different growth rates were demonstrated at pH ranging from 4 to 9, but optimum growth was observed at pH 6 to 8. This is the first record of Haliphthoros milfordensis infection in Malaysia.
  • Osamu Kurata, Shinpei Wada
    FISH & SHELLFISH IMMUNOLOGY 53 110-110 2016年6月  
  • Y. N. Lee, K. Hatai, O. Kurata
    BULLETIN OF THE EUROPEAN ASSOCIATION OF FISH PATHOLOGISTS 36(3) 111-117 2016年  査読有り
    In April 2014, marine Oomycetes were first isolated from mud crab Scylla tranquebarica eggs and larvae at the University Malaysia Sabah shrimp hatchery. A fungus was isolated from infected eggs and larvae using PYGS agar. It was thought that the same fungus infected both eggs and larvae; therefore, strain IPMB 1401 was randomly selected for further characterization in this study. The isolated fungus produced a discharge tube from the mycelium, and a vesicle was formed at the tip. The zoospores swam away after the vesicle separated from the discharge tube. The strain IPMB 1401 was classified as a Lagenidium sp., closely related to L. thermophilum based on the mode of zoospore release. The differences between the strains IPMB 1401 and pathogenic Lagenidium spp. isolated from marine crustaceans were compared in nucleotide sequence of ITS 1 region. As a result, the IPMB 1401 showed high similarity of 99-100% and belonged to the same cluster with L. thermophilum. Therefore, the strain IPMB 1401 was identified as L. thermophilum. This is the first report of Lagenidium infection in Malaysia.
  • Kwanprasert Panchai, Chutima Hanjavanit, Nilubon Rujinanont, Shinpei Wada, Osamu Kurata, Kishio Hatai
    AACL Bioflux 8 70-81 2015年2月14日  
    © 2015 BIOFLUX SRL. All rights reserved. Experimental infection of Nile tilapia (Oreochromis niloticus) fry using 6 Achlya isolates from cultured Nile tilapia with water mold infections was attempted. The experimental fish were exposed to 1.0 x 102 and 1.0 x 104 zoospores mL-1 of each Achlya isolate after ami-momi treatment. The cumulative mortality rates of fish exposed to 1.0 x 104 zoospores mL-1 of A. klebsiana BKKU1003, and A. diffusa BKKU1012 were 88.8 and 77.7%, respectively. A. klebsiana BKKU1003 was more pathogenic than the other isolates. Histopathological examination of the skin of Nile tilapia fry exposed to 1.0 x 102 zoospores mL-1 of A. klebsiana BKKU1003 showed numerous hyphae grew on the skin surface and some areas of skin were sloughed. The fish exposed to 1.0 x 104 zoospores mL-1 of A. klebsiana BKKU1003 showed massive accumulated hyphae on skin lesions with necrosis of the epidermal cells and the hyphae penetrated from the epidermis to the musculature without granulomatous response surrounding the hyphae. We found that it is possible to infect tilapia fry by exposing them to zoospores of Achlya after the ami-momi treatment.
  • Hanako Fukano, Shinpei Wada, Osamu Kurata, Kaori Mizuno, Kazue Nakanaga, Yoshihiko Hoshino
    Fish Pathology 50(2) 68-74 2015年  査読有り
    In 2009 and 2010, unusually high mortality events were recorded among cultured populations of thread-sail filefish Stephanolepis cirrhifer in Ehime Prefecture, Japan. Diseased fish exhibited abdominal distention and many white nodules filled with thick, pale-yellow material scattered on the surface of the serosae of internal organs and mesentery. Histopathologically, the disease was characterized by variable sized granulomatous lesions with central necrotic core surrounded by thin irregular arrangement of epithelioid cells and the outermost thin rim of connective tissue. The central part of granuloma showed colliquative necrosis with abundant cellular debris and some clusters of long-rods that were positive with Ziehl-Neelsen stain. From the results of microbiological examinations and simplified identification with DNA-DNA hybridization, two representative isolates collected in 2009 and 2010 were classified into rapidly growing "nontuberculous mycobacteria (NTM)" that closely related to Mycobacterium chelonae. A pathogenicity test using one of the isolates successfully reproduced the granulomatous lesions closely resembled to those in the spontaneous cases, suggesting that the rapidly growing NTM is pathogenic to the thread-sail filefish, and is a causative agent of the spontaneous case. This is the first report of the disease caused by a rapidly growing NTM in maricultured fish in Japan.
  • Kwanprasert Panchai, Chutima Hanjavanit, Nilubon Rujinanont, Shinpei Wada, Osamu Kurata, Kishio Hatai
    AACL Bioflux 7 529-542 2014年12月21日  
    © 2014, BIOFLUX SRL. All rights reserved. Water mold-infected Nile tilapia (Oreochromis niloticus) from cultured net cages along the Nam Phong River, Khon Kaen Province, northeast Thailand, were collected from September 2010 to August 2011. The 34 obtained water mold isolates belonged to the genus Achlya and were identified as Achlya bisexualis, A. diffusa, A. klebsiana, A. prolifera and unidentified species of Achlya. Isolates of A. bisexualis and A. diffusa were the most abundant (35%), followed by the unidentified species of Achlya (18%) and then, A. klebsiana and A. prolifera (6% each). The ITS1-5.8S-ITS2 region of the unidentified isolates was sequenced for phylogenetic analysis. Three out of 6 isolates were indicated to be A. dubia (BKKU1005), A. bisexualis (BKKU1009 and BKKU1134), and other 3 out of 6 isolates (BKKU1117, BKKU 1118 and BKKU1127) will be an as-yet unidentified species of Achlya. The biological characteristics of the isolates showed optimum temperatures for vegetative growth of 25–35ºC. All of the Achlya isolates were able to grow under up to 1.5% sodium chloride. The isolates grew well, and their zoospores were able to germinate at pH 4.0–11.0. Microscopic examination of the skin lesions of the infected tilapia revealed bacteria and hyphae. Some of the hyphae penetrated into the epidermis, and numerous small blood vessels and scattered macrophages could be observed throughout the infected area.
  • N. Tanaka, T. Izawa, M. Kuwamura, N. Higashiguchi, C. Kezuka, O. Kurata, S. Wada, J. Yamate
    JOURNAL OF FISH DISEASES 37(4) 401-405 2014年4月  査読有り
  • Shinpei Wada, Osamu Kurata, Hitoshi Hatakeyama, Azumi Yamashita, Shusaku Takagi, Toyohiko Nishizawa, Hiroshi Yokoyama
    FISH PATHOLOGY 49(1) 7-15 2014年3月  査読有り
    In 2008, several episodes of mortality were recorded in cultured populations of juvenile greater amberjack reared in the southwest region of Japan. Diseased fish had asymmetrical abdominal distention and pale gills. The head kidney, trunk kidney, and spleen of every fish that was examined was enlarged and discolored. The results of all microbiological and molecular biological assays of tissues taken from diseased fish were negative for major known pathogens. Histopathologically, the disease was characterized by proliferative interstitial nephritis and proliferative splenitis associated with minute, round structures within the cytoplasm of proliferating mono-nucleated cells. Transmission trial using the enlarged trunk kidney from a naturally infected fish successfully reproduced the disease. The results indicate that this disease is caused by an infectious microorganism, and the most likely etiological agent is the minute, round structures which are probably a hitherto unknown eukaryotic microorganism.
  • 山下亜純, 高木修作, 和田新平, 倉田 修, 福田 穣, 平江多績, 中西健二, 黒原健朗
    魚病研究 49(1) 35-38 2014年  査読有り
    2008年と2009年に西日本の主要なカンパチ養殖場において稚魚に腎腫大と脾腫を特徴とする大量死が発生した。そこで,その発生状況を調べ,疫学的特徴を整理した。本症例は,愛媛,鹿児島,宮崎,高知,大分県のカンパチ養殖場において5~9月(水温20~30℃)に中国産輸入種苗で発生した。病魚は重度の貧血や血清K+値および無機リン値の上昇が認められた。大量死と日本国内における養殖業者の飼育管理方法に関連性は認められなかった。一方,大量死は全長15cm以下の輸入種苗の一部のロットに発生し,死亡率は種苗のロットにより大きく異なっていた。特に,輸入後期のロットで死亡率は高く,種苗の履歴との関連性が示唆された。
  • 山下 亜純, 倉田 修, 高木 修作, 和田 新平
    魚病研究 48(2) 63-65 2013年6月15日  
  • O. Kurata, T. Matsuyama, T. Sakai, T. Takano, S. Wada, C. Nakayasu
    FISH & SHELLFISH IMMUNOLOGY 34(6) 1716-1716 2013年6月  
  • 和田新平, 倉田修
    養殖衛生管理問題への調査・研究成果報告書 2011 31,33-43 2012年3月  
  • C. Saejung, K. Hatai, S. Wada, O. Kurata, L. Sanoamuang
    JOURNAL OF FISH DISEASES 34(12) 911-920 2011年12月  
    In this study, black disease infecting fairy shrimps, Streptocephalus sirindhornae Sanoamuang, Murugan, Weekers & Dumont, and Branchinella thailandensis Sanoamuang, Saengphan & Murugan, in Thailand, was investigated. The typical signs of the disease are the appearance of black spots on the cuticle, located mainly on the dorsal side and thoracopods. A number of rod-shaped bacteria aggregated in the black spots and were visualized by scanning electron microscopy. The histopathological results showed that a haemocytic response to the infection resulted in a dense melanized core of bacteria. In addition, generalized septicaemia by rod-shaped bacteria was also observed in the infected tissue. Of the 31 isolates, Aeromonas spp. were predominantly isolated and six strains were selected for the experimental infections. The most pathogenic strain was identified molecularly as A. hydrophila. When fairy shrimp were infected at bacterial concentrations of 10(4) and 10(6) cfu mL(-1), the overall infection levels were 73.33 +/- 6.67% and 93.33 +/- 6.67%, respectively. The experimentally infected fairy shrimp showed abnormal swimming and died within 24-48 h after the appearance of the dark pigment.
  • 和田新平, 倉田修
    養殖衛生管理問題への調査・研究成果報告書 2010 13,15-23 2011年3月  
  • Daisuke Takuma, Ayako Sano, Shinpei Wada, Osamu Kurata, Kishio Hatai
    MYCOSCIENCE 52(2) 119-131 2011年3月  
    A species of Aphanomyces was isolated from juvenile soft-shelled turtles, Pelodiscus sinensis, cultured in Japan. Typically, an infected turtle showed small whitish maculae on the carapace. Many hyphae were observed in the epidermis. The hyphae were isolated using glucose-yeast (GY) agar plates. The morphological characteristics were very similar to those of Aphanomyces laevis, but a clear nuclear spot was observed in the center of the oospore in the strains isolated from the soft-shelled turtles. The optimal growth temperature for the isolates was 25-30A degrees C and the optimum pH was 6-9. Experimental infection tests with isolates produced small whitish maculae on the carapace, and soft-shelled turtles artificially infected with the zoospores showed high mortality, especially in the high-dose group. Phylogenetic analysis based on the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) indicated that the isolates from the soft-shelled turtles were unidentified species of Aphanomyces. As a result, the strain was described as a new species, Aphanomyces sinensis.
  • Shinpei Wada, Hiroko Atami, Osamu Kurata, Kishio Hatai, Kohichi Kasuya, Yusuke Watanabe, Hideo Fukuda
    Fish Pathology 46(2) 59-61 2011年  
    We investigated the histopathological features of the gill lesions of diseased ayu Plecoglossus altivelis that had been diagnosed with 'Boke'. The features were classified into three types: one characterized by atypical gill epithelial cells, one characterized by numerous long Gram-negative rods, and one characterized by both atypical gill epithelial cells and long rods. TEM revealed the presence of poxvirus-like virus in the atypical gill epithelial cells. Our results indicated that 'Boke' was composed of different kinds of diseases. We propose 'atypical cellular gill disease (ACGD)' as the new name of novel gill disease in ayu. © 2011 The Japanese Society of Fish Pathology.
  • Hanjavanit Chutima, Panchai Kwanprasert, 和田 新平, 倉田 修, 畑井 喜司雄
    水産増殖 = The aquiculture 58(4) 533-537 2010年12月20日  
  • Laoprasert Thitiporn, Kanchanakhan Somkiat, 八木田 健司, 和田 新平, 倉田 修, CHINABUT Supranee, 畑井 喜司雄
    水産増殖 = The aquiculture 58(3) 421-424 2010年9月20日  
    原生動物、分子構成、DNA、感染、特性。
  • Pham Minh Duc, Shinpei Wada, Osamu Kurata, Kishio Hatai
    FISH PATHOLOGY 45(3) 109-114 2010年9月  
    Acremonium sp. NJM 0672, isolated from diseased mantis shrimp Oratosquilla oratoria, was susceptible in vitro to three kinds of antifungal agents: voriconazole, amphotericin B and terbinafine hydrochloride. Voriconazole was selected to treat kuruma prawn Penaeus japonicus, which had been intramuscularly injected with 0.1 mL of 5.0 x 10(4) conidia/mL of Acremonium sp. Voriconazole was administered orally at doses of 6 and 2 mg/kg body weight per day for 7 consecutive days, or intramuscularly injected at doses of 4 and 2 mg/kg body weight per day for 3 consecutive days. Both treatments were begun at 6 h after injection of the conidial suspension. The gross features, mortality and histopathological findings demonstrated that voriconazole was an efficient antifungal agent against Acremonium sp.
  • Pham Minh Duc, Shinpei Wada, Osamu Kurata, Kishio Hatai
    FISH PATHOLOGY 45(3) 133-136 2010年9月  
    This study was carried out to assess the pathogenicity of Plectosporium oratosquillae NJM 0662 and Acremonium sp. NJM 0672, both of which were isolated from mantis shrimp Oratosquilla oratoris, to kuruma prawn Penaeus japonicus by intramuscular injection of conidial suspensions. These fungi caused mortality in the injected kuruma prawn. The diseased kuruma prawn showed numerous black spots in the gills. Histopathologically, hyphae in the gill filaments and the injected sites were encapsulated by hemocytes. The results indicate that these two fungi are pathogenic against kuruma prawn.
  • Shinpei Wada, Kazuki Ozeki, Osamu Kurata, Kishio Hatai, Aya Ishida, Kenjiroh Ui
    FISH PATHOLOGY 45(2) 92-95 2010年6月  
    In 2009, a disease with cumulative mortality of 52.5% was observed in a captive population of sailfin sandfish Arctoscopus japonicus reared at an aquarium in Japan. The affected fish had no apparent lesions on the body surfaces, with the exception of skin abrasions on the lower jaw. The trunk kidney was swollen and bore white miliary nodules. The most prominent histopathological feature of the diseased fish was numerous bacterial colonies in the internal organs. However, no inflammatory cellular response was apparent. Single species of bacteria was isolated from the trunk kidney and spleen, and identified as atypical A. salmonicida based on the biochemical and serological characteristics.
  • 和田 新平, 倉田 修, 畑井 喜司雄, 山本 桂子, 高橋 亮太, 小林 利充
    Japanese journal of zoo and wildlife medicine 15(1) 31-36 2010年3月  
    エイ類はその興味深い形態学的特徴と行動から日本国内の水族館で多く展示されている。しかしながら,大型となることと尾部背側に鋭い棘を有することから,彼らの飼育および健康管理には危険が伴う場合が多い。したがって,このような作業を実施するには鎮静・麻酔を施す必要がある。本研究はウシバナトビエイ(Rhinoptera javanica)に対する2-phenoxyethanolの鎮静および麻酔効果について検討することを目的とした。オキナワマリンリサーチセンターで飼育されていたウシバナトビエイ合計12尾を供試し,それぞれ200,400,600ppmとなるように濾過海水で希釈した2-phenoxyethanolに浸漬した。特徴的行動と呼吸回数を5分ごとに観察し,実験終了後には供試薬剤を含まない海水中に移して覚醒までの時間を計測した。それぞれの実験は高水温時および低水温時に2回ずつ実施した。結果より,2-phenoxyethanolを用いてウシバナトビエイを安全に鎮静・麻酔するには,高水温時および低水温時ともに400ppmが最適濃度であると判断された。
  • Takuma Daisuke, Sano Ayako, Wada Shinpei, KURATA Osamu, HATAI Kishio
    Mycoscience 35(6) 432-444 2010年  
  • C. Munchan, O. Kurata, S. Wada, K. Hatai, A. Sano, K. Kamei, N. Nakaoka
    JOURNAL OF FISH DISEASES 32(10) 893-900 2009年10月  
    This report describes Exophiala infection in cultured striped jack, Pseudocaranx dentex, in Japan in 2005. One hundred out of 35 000 fish died per day and mortalities continued for 1 month. Diseased fish showed swelling of the abdomen and kidney distension. Numerous septate hyphae, pale brown in colour, were seen in kidney in squash preparations. Histology revealed abundant fungal hyphae and conidia in gill, heart and kidney. Fungal hyphae were accompanied by cell necrosis and influx of inflammatory, mainly mononuclear cells. The fungus isolated from the diseased fish had septate hyphae, pale brown in colour and 1.8-3.0 mu m in diameter. Conidiogenous cells were conspicuous annellides, short or cylindrical or fusiform in shape. Conidia were one-celled, ellipsoidal with smooth walls, accumulated in balls at the apices of annellides that tended to slide down, 1.5-2.0 mu m in width and 3.0-5.0 mu m in length. The fungus was classified into the genus Exophiala based on its morphology and as Exophiala xenobiotica based on the sequences of the ITS 1-5.8S-ITS 2 regions of rDNA. This is the first record of this fungus in a marine fish.
  • 和田 新平, ウィーラクン ソンポ, 倉田 修, 畑井 喜司雄, 津田 英治
    Japanese journal of zoo and wildlife medicine 14(2) 129-133 2009年9月  
    2004年に姫路市立水族館で飼育中の希少魚種であるニッポンバラタナゴ(Rhodeus ocellatus kurumeus)に死亡するものが認められた。病魚は体表に発赤および潰瘍が観察された。病魚の脾臓には様々な大きさの白色結節が認められ,数尾の魚では腎臓の顕著な腫大も観察された。最も顕著な病理組織学的所見は,頭腎,体腎,脾臓,心臓,腸管,腹膜,肝臓,鰓,躯幹筋,眼および脳にみられた肉芽腫性病変であった。これら肉芽腫性病変は類上皮細胞が渦巻状に配列する構造を呈していた。肉芽腫内には抗酸性を示す長桿菌が多数観察され,病魚から分離された菌株はMycobacterium marinumと同定された。
  • Chutharat Munchan, Osamu Kurata, Shinpei Wada, Kishio Hatai, Noriyoshi Nakaoka, Hidemasa Kawakami
    FISH PATHOLOGY 44(3) 128-132 2009年9月  
    This study compared the histopathology of young striped jack Pseudocaranx dentex experimentally infected with the dematiaceous fungus Ochroconis humicola NJM 0472 with that of spontaneously infected fish. Moribund and freshly dead fish from both groups showed similar histopathology, and appeared to have been killed due to hyphae penetrating the visceral organs. Fish that survived the infection appeared to be able to suppress the fungal growth by well-established inflammatory reaction involving mycotic granulomas and granulation tissues. The results suggested that two types of O. humicola infection occur in young striped jack: an acute type infection, which is characterized by penetrating hyphae that cause direct tissue destruction and a chronic type infection, which is characterized by severe inflammatory reaction that causes functional disorders of the affected organs.
  • Shinpei Wada, Osamu Kurata, Kishio Hatai, Edward J. Noga, Michael J. Dykstra, John S. Burke
    FISH PATHOLOGY 44(3) 151-153 2009年9月  
    Examination via light and electron microscopy of juvenile summer flounder Paralichthys dentatus experiencing a subacute to chronic mortality revealed severe necrotizing hepatitis characterized by necrotic multinucleated giant cells (syncytia). The cytoplasms of syncytia contained paracrystalline arrays of reovirus-like particles, strongly suggesting that the epidemic was caused by the virus. This is the first report of a putative viral infection in summer flounder.
  • Pham Minh Duc, Kishio Hatai, Osamu Kurata, Kozue Tensha, Uchida Yoshitaka, Takashi Yaguchi, Shun-Ichi Udagawa
    MYCOPATHOLOGIA 167(5) 229-247 2009年5月  
    Two fungal pathogens of the mantis shrimp (Oratosquilla oratoria) in Yamaguchi and Aichi Prefectures, Japan are described as the new species Plectosporium oratosquillae and Acremonium sp. (a member of the Emericellopsis marine clade). Both fungi infect the gills of the mantis shrimp, which become brown or black due to melanization. The former species is characterized by its slow growth on artificial seawater yeast extract peptone glucose (PYGS) agar, pale yellow to pale orange or grayish yellow colonies, short cylindrical solitary phialides with a wavy tip, and one-celled ellipsoidal conidia. Although lacking the two-celled conidia demonstrated by the type species Plectosporium tabacinum, the taxonomic placement of the new species was confirmed by DNA sequence analysis of the internal transcribed spacer (ITS) region of ribosomal DNA (ITS1, 5.8S rDNA and ITS2). Acremonium sp., the other causal pathogen, differs from P. oratosquillae by its fast growth on PYGS agar, pale orange to salmon-colored colonies, long, slender conidiophores consisting of solitary phialides with tips lacking an undulate outline, and typically cylindrical conidia. Analysis of ITS and beta-tubulin gene sequences placed this fungus within the phylogenetically distinct Emericellopsis (anam. Acremonium) marine clade. Various physiological characteristics of both pathogens were also investigated. This is the first report of a fungal infection found on the mantis shrimp in Japan.
  • 北本恵理, 小谷知也, 倉田修, 伏見浩, 畑井喜司雄, 佐藤秀一
    福山大学内海生物資源研究所報告 (19) 44-45 2009年2月28日  
  • 神嵜健太, 藤加菜子, 長谷川理, 倉田修, 河合純, 林崎良英, 坂本崇, 岡本信明
    動物遺伝育種研究 36(2) 219 2008年12月1日  
  • Masashi Yokota, Seiichi Watanabe, Kishio Hatai, Osamu Kurata, Mituru Furihata, Takahiko Usui
    JOURNAL OF AQUATIC ANIMAL HEALTH 20(4) 207-214 2008年12月  
    The epidemic process of the parasite Ichthyophonus hoferi in cultured rainbow trout Oncorhynchus mykiss was quantitatively estimated by both the cohabitation experiment and two standard models (the Kermarck-McKendrick model and the Reed-Frost model). For analysis of the parasite transmission by cohabitation, fish in two replicate tanks were exposed to 1, 5, or 10 infected fish, and daily mortality was counted for 102 d. Despite simple experiments for artificial exposure to the pathogen, the daily estimate of dead fish in the Kermarck-McKendrick model did not fit the observed number of dead fish in the experiment. In contrast, when the longest possible incubation period (generation time) wag assumed to be 51 d in the Reed-Frost model, the estimated number of dead fish in discrete generations was close to the observed number of dead fish. If the time unit was 51 d, the estimated mortalities in the generation-based Kermarck-McKendrick model were significantly correlated with observed mortalities. These results suggest that the deterministic aspects of the epidemic process of the parasite can be quantitatively deomonstrated on a 51-d timescale or longer, whereas transmission on a daily timescale is uncertain.
  • 小谷 知也, 北本 恵理, 倉田 修, 平山 敬大, 伏見 浩, 佐藤 秀一, 畑井 喜司雄, 宮嶋 暁, 能味 堂郎, 嶋田 貴志
    水産増殖 = The aquiculture 56(3) 375-382 2008年9月20日  
    腸球菌Enterococcus faecalis FK-23加熱死菌体(FK-23)添加配合飼料を給餌し、ワクチンを接種した時のトラフグおよびヒラメの特異免疫能向上について検討した。FK-23添加濃度を0〜1.25%の5段階に調節した配合飼料を用意した。ワクチンとしてVibrio anguillarumホルマリン不活化菌体を2度接種し、1度目の接種後から毎週血清を採取し、血清中抗体のV. anguillarum細胞への凝集能を観察した。トラフグでは、1.25%添加飼料給餌区で凝集能が最も上昇したが、成長しなかった。ヒラメでは抗体価の上昇を確認できなかった。特に0.01%以上では上昇せず、特異的免疫能よりも非特異的免疫能が活性化されたために起こった現象であると考えられた。以上より、FK-23添加飼料給餌はワクチン効果の上昇とその持続を可能にするが、非特異的免疫能の活性化も示唆された。
  • Shinpei Wada, Osamu Kurata, Kishio Hatai, Hideo Ishii, Kohichi Kasuya, Yuhsuke Watanabe
    FISH PATHOLOGY 43(2) 89-91 2008年6月  
    In 1998 and 1999, severe episodes of mortality, often reaching 90%, were recorded among cultured populations of ayu Plecoglossus altivelis reared in Japan. The diseased fish showed appetite reduction and abnormal swimming behavior. Histopathological examination revealed proliferative branchitis with enlarged and atypical epithelial cells. Abundant electron-dense, virus-like particles were observed within the cells under transmission electron microscopy. The particles had a cocoon-like shape and ranged in length from 200 to 300 nm, indicating a member of the poxvirus group. These findings suggest the possibility that the mortality events are related to infection of a poxvirus-like virus.

書籍等出版物

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講演・口頭発表等

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担当経験のある科目(授業)

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共同研究・競争的資金等の研究課題

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産業財産権

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