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2. Non-tuberculous mycobacterial disease associated with Mycobacterium montefiorense in salamanders

Author

Komine, Takeshi, primary, Ihara, Hyogo, additional, Inohana, Mari, additional, Kwok, Jennifer Caroline, additional, Shimizu, Akane, additional, Terasawa, Tsumugi, additional, Miyazaki, Ayaka, additional, Srivorakul, Saralee, additional, Iwao, Hajime, additional, Harada, Sachiko, additional, Yoshida, Mitsunori, additional, Hoshino, Yoshihiko, additional, Kurata, Osamu, additional, Fukano, Hanako, additional, and Wada, Shinpei, additional

Published
2023
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7. Confirmation of a Suspected Case of paracoccidioidomycosis Ceti by a Combination of Polymerase Chain Reaction and Loop-mediated Isothermal Amplification (PCR-LAMP) Analysis

Author

Kanegae, Hikaru, Suguiura, Massahiro de Souza Igor, Minakawa, Tomoko, Augusuto, Mario Ono, Itano, Nakagwa Eiko, Wada, Shinpei, Nakamura, Yuichi, Shumoto, Godai, Sano, Ayako, and Ueda, Keiichi

Subjects
クジラ型パラコクシジオイデス症, Diagnoses, dolphin, PCR-LAMP, LAMP 法, paracoccidoioidomycosis ceti, 診断, loop-mediated isothermal amplification (LAMP), カマイルカ, Paracoccidioidomycosis ceti
Abstract

Paracoccidioidomycosis ceti (PCM-C) is a zoonotic mycosis characterized by chronic granulomatous keloidal dermatitis in cetaceans that has been reported worldwide. The causative agents of PCM-C are unculturable Paracoccidioides brasiliensis var. ceti and Paracoccidioides app., which are genetically identical to one of the causative agents of paracoccidioidomycosis: P. brasiliensis sensu stricto. The definitive diagnosis of PCM-C is based on clinical symptoms and the detection of yeast-like cells during pathological examinations; molecular diagnosis is not essential. However, analyses at the molecular level are important to distinguish PCM-C from other fungal infections showing similar clinical signs. Researchers have been following a suspected case of PCM-C in a Pacific white-sided dolphin (Lagenorhynchus obliquidens) by performing cytology, histopathology, and molecular biology experiments since 2015. Finally, we confirmed it as the fourth Japanese PCM-C case based on the partial sequence of 43-kDa glycoprotein antigen gene (gp43) with 99.4% identity to that from P. brasiliensis sensu stricto from a biopsied sample in October 2019. Then, we applied a loop-mediated isothermal amplification (LAMP) for P. brasiliensis. The simple LAMP targeting for fungal DNA and human clinical materials failed to amplify bands, but a combination of polymerase chain reaction (PCR) and LAMP (PCR-LAMP) could amplify species-specific ladder-like bands identical to P. brasiliensis., クジラ型パラコクシジオイデス症 (paracoccidoioidmycosis ceti : PCM-C) は，イルカを宿主とし，難治性慢性肉芽腫性ケロイド状皮膚炎を特徴とする人獣共通真菌症である。原因菌は非培養性の Paracoccidioides brasiliensis var. ceti で，中南米を流行地とする高度病原性真菌症のパラコクシジオイデス症 (PCM) の原因菌 P. brasiliensis と遺伝子型は同一である。確定診断は臨床症状と病理像での酵母細胞の証明であるが，遺伝子情報による診断も重要である。今回，遺伝子情報を欠くものの， PCM-C が疑われていたイルカ皮膚病変生検組織より nested-PCR で原因菌の特異的糖タンパク抗原遺伝子である gp43 が検出され，配列は既報の PCM-C 国内第３症例目と98.9% 相同であった。そこで PCM の診断用に設計された LAMP 法を適用したところ， PCR と LAMP 法の組み合わせにより gp43 の増幅に成功したところから，この手法は迅速診断法としての有用性が期待できる。

Published
2021

18. Genital mycobacteriosis caused by Mycobacterium marinum detected in two captive sharks by peptide nucleic acid–fluorescence in situ hybridization.

Author

Inohana, Mari, Komine, Takeshi, Tanaka, Yoshiaki, Kurata, Osamu, and Wada, Shinpei

Subjects
IN situ hybridization, MYCOBACTERIOSIS, PEPTIDES, MYCOBACTERIUM, NUCLEIC acids, CHONDRICHTHYES, SHARKS, POLYMERASE chain reaction
Abstract

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. [ABSTRACT FROM AUTHOR]

Published
2023
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28. Development of New PCR and Quantitative PCR Protocols for the Detection of Plecoglossus altivelis Poxvirus-like Virus in Atypical Cellular Gill Disease of Ayu

Author

Koyama, Tomoki, primary, Komatsu, Daiki, additional, Uchino, Tsubasa, additional, Midorikawa, Yuki, additional, Kato, Goshi, additional, Ishikawa, Takanori, additional, Nishimura, Tomohiro, additional, Takeda, Korenori, additional, Fukuda, Hideo, additional, Wada, Shinpei, additional, and Sano, Motohiko, additional

Published
2020
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42. Supplementary data for the article: Beškoski, V.; Yamamoto, K.; Yamamoto, A.; Okamura, H.; Hayashi, M.; Nakano, T.; Matsumura, C.; Fukushi, K.; Wada, S.; Inui, H. Distribution of Perfluoroalkyl Compounds in Osaka Bay and Coastal Waters of Western Japan. Chemosphere 2017, 170, 260–265. https://doi.org/10.1016/j.chemosphere.2016.12.028

Author

Beškoski, Vladimir, Yamamoto, Katsuya, Yamamoto, Atsushi, Okamura, Hideo, Hayashi, Mitsuru, Nakano, Takeshi, Matsumura, Chisato, Fukushi, Keiichi, Wada, Shinpei, Inui, Hideyuki, Beškoski, Vladimir, Yamamoto, Katsuya, Yamamoto, Atsushi, Okamura, Hideo, Hayashi, Mitsuru, Nakano, Takeshi, Matsumura, Chisato, Fukushi, Keiichi, Wada, Shinpei, and Inui, Hideyuki

Published
2017

43. Distribution of perfluoroalkyl compounds in Osaka Bay and coastal waters of Western Japan

Author

Beškoski, Vladimir, Yamamoto, Katsuya, Yamamoto, Atsushi, Okamura, Hideo, Hayashi, Mitsuru, Nakano, Takeshi, Matsumura, Chisato, Fukushi, Keiichi, Wada, Shinpei, Inui, Hideyuki, Beškoski, Vladimir, Yamamoto, Katsuya, Yamamoto, Atsushi, Okamura, Hideo, Hayashi, Mitsuru, Nakano, Takeshi, Matsumura, Chisato, Fukushi, Keiichi, Wada, Shinpei, and Inui, Hideyuki

Abstract

Perfluoroalkyl acids (PFAAs) including perfluoroalkyl sulfonates (PFSAs) and perfluoroallcyl carboxylates (PFCAs) were analyzed in sediment samples taken from Ajifu Waterway in Osaka city, from Osaka Bay, and from Kagoshima Bay, as well as in fifteen seawater samples collected from Osaka Bay and coastal waters of Western Japan. In all sediment samples, only PFCAs were detected, and the highest concentration was determined in Ajifu Waterway, where Sigma PFAA was 58990 ng kg(-1) dry weight. The total concentrations of PFAAs in sea water samples ranged between the limit of quantification and 53.4 ng L-1, and perfluorohexanoic acid was the most prevalent and had the highest concentration of 37 ng L-1. The changes in the patterns and concentrations of PFAA5 in Osaka Bay and coastal waters of Western Japan indicate that the PFAAs in surface waters are influenced by sources from Keihanshin Metropolitan Area, mainly the Yodo River basin, and the dilution effect which naturally occurs during their transport to the Pacific Ocean. (C) 2016 Elsevier Ltd. All rights reserved.

Published
2017

48. Experimental pathogenicity of Achlya species from cultured Nile tilapia to Nile tilapia fry in Thailand

Author

Hatai, Kishio, Panchai, Kwanprasert, Hanjavanit, Chutima, Rujinanont, Nilubon, Wada, Shinpei, Kurata, Osamu, Hatai, Kishio, Panchai, Kwanprasert, Hanjavanit, Chutima, Rujinanont, Nilubon, Wada, Shinpei, and Kurata, Osamu

Abstract

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. © 2015 BIOFLUX SRL. All rights reserved.

Published
2015

50. In vitro antifungal activity of Cassia fistula L. against selected pathogenic water molds

Author

Borisutpeth, Manassanan; Faculty of Veterinary Medicine, Khon Kaen University, Khon Kaen 40002, Thailand, Kanbutra, Pithai; Faculty of Veterinary Medicine, Khon Kaen University, Khon Kaen 40002, Thailand, Weerakhun, Sompoth; Faculty of Veterinary Medicine, Khon Kaen University, Khon Kaen 40002, Thailand, Wada, Shinpei; Laboratory of Aquatic Medicine, Nippon Veterinary and Life Science University, Musashino, Tokyo 180-8602, Japan, Hatai, Kishio; Laboratory of Aquatic Medicine, Nippon Veterinary and Life Science University, Musashino, Tokyo 180-8602, Japan, Borisutpeth, Manassanan; Faculty of Veterinary Medicine, Khon Kaen University, Khon Kaen 40002, Thailand, Kanbutra, Pithai; Faculty of Veterinary Medicine, Khon Kaen University, Khon Kaen 40002, Thailand, Weerakhun, Sompoth; Faculty of Veterinary Medicine, Khon Kaen University, Khon Kaen 40002, Thailand, Wada, Shinpei; Laboratory of Aquatic Medicine, Nippon Veterinary and Life Science University, Musashino, Tokyo 180-8602, Japan, and Hatai, Kishio; Laboratory of Aquatic Medicine, Nippon Veterinary and Life Science University, Musashino, Tokyo 180-8602, Japan

Abstract

Water mold infections in both cultured and wild aquatic animals, caused by members of the genera Saprolegnia, Achlya, and Aphanomyces, have widely occurred worldwide. Outbreaks in aquatic creatures, especially in aquacultural facilities and fish hatcheries are common problem. Cassia fistula L. has been used as medicinal plant for broad purposes including for fungal infectious diseases remedy, but efficacy against water molds is still not apparently known. The present study was aimed to investigate in vitro antifungal activity of C. fistula stem-bark alcoholic extracts against the selected pathogenic water molds isolated from fish. The results showed that the C. fistula extract was capable to kill hyphae and zoospores of Saprolegnia paracitica NJM 8604 and S. diclina NJM 0005 at levels ranging from 1,000-4,000 microg/mL evidently within 24 h-exposure, while, a level of 500 microg/mL was sufficient to kill the both stages of Aphanomyces invadans NJM 9701. In addition, the antifungal activity of C. fistula absolute alcoholic extract comparing to 50% aqueous alcoholic extract were seemingly not different. Conclusively, the C. fistula stem-bark extracts, both absolute and 50% aqueous alcoholic extractions, were effectively able to inhibit growth and kill the 3 strains of pathogenic water molds by affect throughout the 2 important, zoospore and hyphal, stages.

Published
2014

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