T. Takano, T. Matsuyama, Y. Kawato, T. Sakai, J. Kurita, Yuta Matsuura, Sachiko Terashima, K. Nakajima, C. Nakayasu
Red sea bream iridoviral disease (RSIVD) spreads readily throughout the marine environment because of its wide variety of host species. Use of a rapid diagnostic method, such as the immunofluorescence antibody test, helps to control the disease, so the anti-RSIV monoclonal antibody M10 (mAb M10) has been developed in Japan. In the present study, we carried out epitope mapping using the phage display method to identify the antigen of mAb M10. A phage display RSIV peptide library was constructed to cover the entire genome of RSIV, then phage clones recognized by the antibody were selected by biopanning. The selected clones harbored partial fragments of the laminin-type epidermal growth factor-like domain (LEGFD) gene. N-terminal and C-terminal deletion peptides were then prepared from the amino acid sequence deduced from the smallest fragment to precisely determine the epitope. Finally, seven amino acids, EYDCPEY, located in the extracellular domain of the LEGFD protein were determined to be the epitope. Identical residues of the epitope were also identified from the LEGFD protein in other megalocytiviruses including the infectious spleen and kidney necrosis virus and turbot reddish body iridovirus. mAb M10 is considered to be widely available for the diagnostics of megalocytivirus infections.
{"title":"Identification of the Epitope Recognized by the Anti-Red Sea Bream Iridovirus (RSIV) Monoclonal Antibody M10 Using a Phage Display RSIV Peptide Library","authors":"T. Takano, T. Matsuyama, Y. Kawato, T. Sakai, J. Kurita, Yuta Matsuura, Sachiko Terashima, K. Nakajima, C. Nakayasu","doi":"10.3147/jsfp.54.83","DOIUrl":"https://doi.org/10.3147/jsfp.54.83","url":null,"abstract":"Red sea bream iridoviral disease (RSIVD) spreads readily throughout the marine environment because of its wide variety of host species. Use of a rapid diagnostic method, such as the immunofluorescence antibody test, helps to control the disease, so the anti-RSIV monoclonal antibody M10 (mAb M10) has been developed in Japan. In the present study, we carried out epitope mapping using the phage display method to identify the antigen of mAb M10. A phage display RSIV peptide library was constructed to cover the entire genome of RSIV, then phage clones recognized by the antibody were selected by biopanning. The selected clones harbored partial fragments of the laminin-type epidermal growth factor-like domain (LEGFD) gene. N-terminal and C-terminal deletion peptides were then prepared from the amino acid sequence deduced from the smallest fragment to precisely determine the epitope. Finally, seven amino acids, EYDCPEY, located in the extracellular domain of the LEGFD protein were determined to be the epitope. Identical residues of the epitope were also identified from the LEGFD protein in other megalocytiviruses including the infectious spleen and kidney necrosis virus and turbot reddish body iridovirus. mAb M10 is considered to be widely available for the diagnostics of megalocytivirus infections.","PeriodicalId":51052,"journal":{"name":"Fish Pathology","volume":"14 1","pages":""},"PeriodicalIF":0.6,"publicationDate":"2020-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83515188","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
― Recently, our lab reported that CUBIC, a tissue-clearing technique, could help reveal the initial route of infection, its spread, and the localization of pathogens in infected goldfish Carassius auratus . However, this tech-nique, in its original form, failed to clear melanin pigmented parts of the fish. We show here that bleaching with H 2 O 2 clears melanin pigments of Black Moor goldfish without causing severe histological damage. Furthermore, nuclear staining with PI helps visualize the internal structures of cleared fish in a dark field. Since bleaching does not sig-nificantly quench the fluorescence of GFP, it may be applied to the in vivo imaging of GFP.
{"title":"Application of Hydrogen Peroxide-Melanin Bleaching and Fluorescent Nuclear Staining for Whole-Body Clearing and Imaging in Fish","authors":"K. Futami, Oto Furukawa, M. Maita, T. Katagiri","doi":"10.3147/jsfp.54.101","DOIUrl":"https://doi.org/10.3147/jsfp.54.101","url":null,"abstract":"― Recently, our lab reported that CUBIC, a tissue-clearing technique, could help reveal the initial route of infection, its spread, and the localization of pathogens in infected goldfish Carassius auratus . However, this tech-nique, in its original form, failed to clear melanin pigmented parts of the fish. We show here that bleaching with H 2 O 2 clears melanin pigments of Black Moor goldfish without causing severe histological damage. Furthermore, nuclear staining with PI helps visualize the internal structures of cleared fish in a dark field. Since bleaching does not sig-nificantly quench the fluorescence of GFP, it may be applied to the in vivo imaging of GFP.","PeriodicalId":51052,"journal":{"name":"Fish Pathology","volume":"2008 1","pages":""},"PeriodicalIF":0.6,"publicationDate":"2020-01-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82582708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
― We examined how mechanical damages to the skin and the infestation of the monogenean Benedenia seriolae on the skin affected Nocardia seriolae infection in Japanese amberjack Seriola quinqueradiata . After the skin was inflicted damages or after B. seriolae was dewormed with freshwater bathing, amberjacks were challenged with 1.0 × 10 3 CFU/mL N. seriolae by immersion. Survival rate was significantly lower in the skin-damaged group and the skin fluke-infested group than in their respec-tive control groups. The present study suggests that skin injuries and B. seriolae infection contribute to the N. seriolae infection in Japanese amberjack.
{"title":"Skin Injuries Contribute to Nocardiosis in Japanese Amberjack Seriola quinqueradiata","authors":"Y. Miyoshi, Y. Fukuda, K. Ogawa","doi":"10.3147/jsfp.54.64","DOIUrl":"https://doi.org/10.3147/jsfp.54.64","url":null,"abstract":"― We examined how mechanical damages to the skin and the infestation of the monogenean Benedenia seriolae on the skin affected Nocardia seriolae infection in Japanese amberjack Seriola quinqueradiata . After the skin was inflicted damages or after B. seriolae was dewormed with freshwater bathing, amberjacks were challenged with 1.0 × 10 3 CFU/mL N. seriolae by immersion. Survival rate was significantly lower in the skin-damaged group and the skin fluke-infested group than in their respec-tive control groups. The present study suggests that skin injuries and B. seriolae infection contribute to the N. seriolae infection in Japanese amberjack.","PeriodicalId":51052,"journal":{"name":"Fish Pathology","volume":"22 1","pages":""},"PeriodicalIF":0.6,"publicationDate":"2019-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79367649","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hisato Takeuchi, D. Nakano, Mayu Iwadare, Ryuji Tomono, T. Imai, Kazutomo Hori, Hiroaki Izumi, A. Namba, Sonoko Shimizu, T. Nakai, N. Mano
Experimental infection trials of six riverine fish species with Edwardsiella ictaluri were performed using an immersion method at three water temperatures, 20°C, 25°C, and 30°C. At 20°C, mortalities were recorded only in ayu Plecoglossus altivelis and amur catfish Silurus asotus. Whereas, at 25°C or 30°C, mortality events occurred in other species including Japanese dace Tribilodon hakonensis, gibachi Tachysurus tokiensis, and Japanese eel Anguilla japonica, except for common carp Cyprinus carpio, and high (≥80%) cumulative mortalities were recorded in ayu, amur catfish, and gibachi. These results suggest that many riverine fishes are at risk of E. ictaluri infection during periods of high water temperature.
{"title":"Evaluation of Pathogenicity of Edwardsiella ictaluri to Riverine Fish Species Using an Immersion Infection Method","authors":"Hisato Takeuchi, D. Nakano, Mayu Iwadare, Ryuji Tomono, T. Imai, Kazutomo Hori, Hiroaki Izumi, A. Namba, Sonoko Shimizu, T. Nakai, N. Mano","doi":"10.3147/jsfp.54.61","DOIUrl":"https://doi.org/10.3147/jsfp.54.61","url":null,"abstract":"Experimental infection trials of six riverine fish species with Edwardsiella ictaluri were performed using an immersion method at three water temperatures, 20°C, 25°C, and 30°C. At 20°C, mortalities were recorded only in ayu Plecoglossus altivelis and amur catfish Silurus asotus. Whereas, at 25°C or 30°C, mortality events occurred in other species including Japanese dace Tribilodon hakonensis, gibachi Tachysurus tokiensis, and Japanese eel Anguilla japonica, except for common carp Cyprinus carpio, and high (≥80%) cumulative mortalities were recorded in ayu, amur catfish, and gibachi. These results suggest that many riverine fishes are at risk of E. ictaluri infection during periods of high water temperature.","PeriodicalId":51052,"journal":{"name":"Fish Pathology","volume":"70 1","pages":""},"PeriodicalIF":0.6,"publicationDate":"2019-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85077879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
― Flavobacterium psychrophilum , the causative agent of bacterial cold-water disease, is classified into two genotypes (A and B) based on peptidyl-prolyl cis-trans isomerase C (PPIC) gene polymorphism. These are routinely detected using restriction fragment length polymorphism of the PCR amplicon (PCR-RFLP). Since this method is difficult and time-consuming, a simple method is required. Here we developed a new method using real-time PCR assay (TaqMan probe assay) to detect single nucleotide polymorphism (SNP) in PPIC region. We designed the probes on a SNP site and performed TaqMan probe genotyping. The results obtained using this method was in line with those of RFLP. This new method could easily and rapidly determine F. psychrophilum genotypes.
{"title":"Development of a Real-time PCR to Detect Genotypes A and B in Flavobacterium psychrophilum","authors":"Ryo Inoue, T. Takase","doi":"10.3147/jsfp.54.58","DOIUrl":"https://doi.org/10.3147/jsfp.54.58","url":null,"abstract":"― Flavobacterium psychrophilum , the causative agent of bacterial cold-water disease, is classified into two genotypes (A and B) based on peptidyl-prolyl cis-trans isomerase C (PPIC) gene polymorphism. These are routinely detected using restriction fragment length polymorphism of the PCR amplicon (PCR-RFLP). Since this method is difficult and time-consuming, a simple method is required. Here we developed a new method using real-time PCR assay (TaqMan probe assay) to detect single nucleotide polymorphism (SNP) in PPIC region. We designed the probes on a SNP site and performed TaqMan probe genotyping. The results obtained using this method was in line with those of RFLP. This new method could easily and rapidly determine F. psychrophilum genotypes.","PeriodicalId":51052,"journal":{"name":"Fish Pathology","volume":"31 1","pages":""},"PeriodicalIF":0.6,"publicationDate":"2019-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73256276","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yin-Ze Shi, I. Nishiki, S. Yanagi, Terutoyo Yoshida
Lactococcus garvieae serotype II, a newly emerging causal pathogen of lactococcal infection, has spread to fish farms since 2012, and its damage to aquaculture has increased. A total of 161 strains were isolated from four fish species, yellowtail Seriola quinqueradiata, amberjack S. dumerili, kingfish S. lalandi, and striped jack Pseudocaranx dentex, in eight prefectures between 2012 and 2017. This is the first epidemiological study on L. garvieae serotype II, by using agglutination titers against antisera for diagnosis, biased sinusoidal field gel electrophoresis (BSFGE) analysis, and antimicrobial susceptibility test. Almost none of these strains were agglutinated with anti-Ia serum (anti-KG serum), except for two strains agglutinated with both anti-Ia and II sera. BSFGE analysis revealed that genetically homogeneous L. garvieae serotype II has spread to fish farms since the first outbreak in 2012, and the strains resistant to lincomycin were already found in 2015.
garvieae乳球菌血清型(Lactococcus garvieae serotype II)是一种新出现的乳球菌感染病原，自2012年开始在养鱼场蔓延，对水产养殖造成的危害越来越大。2012 - 2017年，在中国8个县共从quinqueradiata黄尾鱼、amberjack S. dumerili、kingfish S. lalandi和striped jack Pseudocaranx dentex 4种鱼类中分离到161个菌株。本文首次采用抗血清凝集滴度、偏正弦场凝胶电泳(BSFGE)分析和药敏试验对鸡乳杆菌II型进行流行病学研究。除2株菌株同时与抗ia和抗II血清凝集外，其余菌株几乎不与抗ia血清(抗kg血清)凝集。BSFGE分析显示，自2012年首次暴发以来，遗传同质的鸡乳杆菌血清II型已传播到养鱼场，2015年已发现对林可霉素耐药的菌株。
{"title":"Epidemiological Study on Newly Emerging Lactococcus garvieae Serotype II Isolated from Marine Fish Species in Japan","authors":"Yin-Ze Shi, I. Nishiki, S. Yanagi, Terutoyo Yoshida","doi":"10.3147/jsfp.54.51","DOIUrl":"https://doi.org/10.3147/jsfp.54.51","url":null,"abstract":"Lactococcus garvieae serotype II, a newly emerging causal pathogen of lactococcal infection, has spread to fish farms since 2012, and its damage to aquaculture has increased. A total of 161 strains were isolated from four fish species, yellowtail Seriola quinqueradiata, amberjack S. dumerili, kingfish S. lalandi, and striped jack Pseudocaranx dentex, in eight prefectures between 2012 and 2017. This is the first epidemiological study on L. garvieae serotype II, by using agglutination titers against antisera for diagnosis, biased sinusoidal field gel electrophoresis (BSFGE) analysis, and antimicrobial susceptibility test. Almost none of these strains were agglutinated with anti-Ia serum (anti-KG serum), except for two strains agglutinated with both anti-Ia and II sera. BSFGE analysis revealed that genetically homogeneous L. garvieae serotype II has spread to fish farms since the first outbreak in 2012, and the strains resistant to lincomycin were already found in 2015.","PeriodicalId":51052,"journal":{"name":"Fish Pathology","volume":"24 1","pages":""},"PeriodicalIF":0.6,"publicationDate":"2019-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90359966","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"A Survey of Water Temperature for Production of Juvenile Ayu Plecoglossus altivelis","authors":"","doi":"10.3147/jsfp.54.40","DOIUrl":"https://doi.org/10.3147/jsfp.54.40","url":null,"abstract":"","PeriodicalId":51052,"journal":{"name":"Fish Pathology","volume":"4 1","pages":""},"PeriodicalIF":0.6,"publicationDate":"2019-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74336456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
― In a previous field survey of Perkinsus olseni in Manila clams in a tidal flat contaminated with the parasite, the parasite was not detected in spats smaller than 2 mm in shell length collected between April and June. To under-stand the reason, we undertook an experimental challenge, exposing clam spats to the zoospores of parasite. The spats became infected and began to die when mean infection intensity reached about 10 6 cells/g soft tissue. Our results, in conjunction with previously reported filtration rates of Manila clams, suggest the absence of infected spats was attributable to clams’ low filtration and low zoospore densities.
{"title":"Mortality of Spats of Manila Clam Ruditapes philippinarum Experimentally Challenged with the Protozoan Parasite Perkinsus olseni","authors":"Tsukasa Waki, T. Yoshinaga","doi":"10.3147/JSFP.54.34","DOIUrl":"https://doi.org/10.3147/JSFP.54.34","url":null,"abstract":"― In a previous field survey of Perkinsus olseni in Manila clams in a tidal flat contaminated with the parasite, the parasite was not detected in spats smaller than 2 mm in shell length collected between April and June. To under-stand the reason, we undertook an experimental challenge, exposing clam spats to the zoospores of parasite. The spats became infected and began to die when mean infection intensity reached about 10 6 cells/g soft tissue. Our results, in conjunction with previously reported filtration rates of Manila clams, suggest the absence of infected spats was attributable to clams’ low filtration and low zoospore densities.","PeriodicalId":51052,"journal":{"name":"Fish Pathology","volume":"35 1","pages":""},"PeriodicalIF":0.6,"publicationDate":"2019-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84537184","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
S. Kawato, Keigo Kobayashi, S. Shirakashi, S. Yanagi, Y. Fukuda, H. Yamashita, R. Nozaki, I. Hirono, H. Kondo
― Infection with the skin fluke Benedenia seriolae is a serious threat of yellowtail aquaculture. Development of genomic resources and molecular markers for B. seriolae is an essential infrastructure to develop control strategies against B. seriolae infections. As a seminal study to establish the genomic resources of the Japanese B. seriolae populations, we sequenced the complete mitochondrial genome (mitogenome) sequences of 12 B. seriolae specimens collected from four prefectures in Japan. The Japanese B. seriolae mitogenomes were mutually 99% identical, while the identity was 85% when compared to that of an Australian specimen. Furthermore, the gene arrangement in the Japanese B. seriolae mitogenome was slightly different from that in the Australian specimen. The substantial mitogenomic divergence between the Australian and Japanese specimens indicates that B. seriolae is composed of molecularly divergent populations. Variable sites were dispersed in the mitogenome of the Japanese B. seriolae . Phylogenetic relationships among the Japanese B. seriolae specimens did not reflect the host species or geographic distributions within Japan.
{"title":"Phylogenetic Analysis with Complete Mitochondrial Genome Sequences of Benedenia seriolae Specimens Derived from Japanese Seriola spp.","authors":"S. Kawato, Keigo Kobayashi, S. Shirakashi, S. Yanagi, Y. Fukuda, H. Yamashita, R. Nozaki, I. Hirono, H. Kondo","doi":"10.3147/JSFP.54.27","DOIUrl":"https://doi.org/10.3147/JSFP.54.27","url":null,"abstract":"― Infection with the skin fluke Benedenia seriolae is a serious threat of yellowtail aquaculture. Development of genomic resources and molecular markers for B. seriolae is an essential infrastructure to develop control strategies against B. seriolae infections. As a seminal study to establish the genomic resources of the Japanese B. seriolae populations, we sequenced the complete mitochondrial genome (mitogenome) sequences of 12 B. seriolae specimens collected from four prefectures in Japan. The Japanese B. seriolae mitogenomes were mutually 99% identical, while the identity was 85% when compared to that of an Australian specimen. Furthermore, the gene arrangement in the Japanese B. seriolae mitogenome was slightly different from that in the Australian specimen. The substantial mitogenomic divergence between the Australian and Japanese specimens indicates that B. seriolae is composed of molecularly divergent populations. Variable sites were dispersed in the mitogenome of the Japanese B. seriolae . Phylogenetic relationships among the Japanese B. seriolae specimens did not reflect the host species or geographic distributions within Japan.","PeriodicalId":51052,"journal":{"name":"Fish Pathology","volume":"26 1","pages":""},"PeriodicalIF":0.6,"publicationDate":"2019-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80173504","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
― A novel PCR-RFLP typing method was developed using PCR primers specific for the B subunit of DNA gyrase gene ( gyrB ) of Flavobacterium psychrophilum and restriction endonuclease Bsp 119I. With this PCR-RFLP, 300 isolates of F. psychrophilum were classified into two genotypes (197 and 103 isolates), and the respective genotypes were correlated with the host fish species from which the isolates were derived. These isolates were further subdivided into 12 genotypes by combining with three previous PCR-RFLPs. It is suggested that this PCR-RFLP typing system can be a useful tool in epizootiological studies on bacterial cold-water disease.
{"title":"A Novel PCR-RFLP Genotyping of Flavobacterium psychrophilum Targeting the gyrB Region","authors":"S. Izumi, H. Arai, Kyuma Suzuki, F. Aranishi","doi":"10.3147/JSFP.54.37","DOIUrl":"https://doi.org/10.3147/JSFP.54.37","url":null,"abstract":"― A novel PCR-RFLP typing method was developed using PCR primers specific for the B subunit of DNA gyrase gene ( gyrB ) of Flavobacterium psychrophilum and restriction endonuclease Bsp 119I. With this PCR-RFLP, 300 isolates of F. psychrophilum were classified into two genotypes (197 and 103 isolates), and the respective genotypes were correlated with the host fish species from which the isolates were derived. These isolates were further subdivided into 12 genotypes by combining with three previous PCR-RFLPs. It is suggested that this PCR-RFLP typing system can be a useful tool in epizootiological studies on bacterial cold-water disease.","PeriodicalId":51052,"journal":{"name":"Fish Pathology","volume":"25 1","pages":""},"PeriodicalIF":0.6,"publicationDate":"2019-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83024056","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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