Giuseppe Esposito, Stefania Bergagna, Silvia Colussi, Khalid Shahin, Roberta Rosa, Donatella Volpatti, Caterina Faggio, Camilla Mossotto, Alice Gabetti, Alessandra Maganza, Elena Bozzetta, Marino Prearo, Paolo Pastorino
The aquaculture sector plays a vital role in global food security, yet it grapples with significant challenges posed by infectious diseases. Piscine lactococcosis is one of the significant threats in rainbow trout aquaculture due to its potential to cause severe economic losses through mortalities, reduced growth rates, and increased susceptibility to other pathogens. It poses challenges in disease management strategies, impacting the sustainability and profitability of rainbow trout farming. The current study focuses on the variations in serum blood parameters of farmed rainbow trout Oncorhynchus mykiss during a lactococcosis outbreak caused by Lactococcus garvieae. Blood samples were collected for biochemical analysis, fish were examined for parasites and bacteria, and DNA from bacterial colonies was PCR-amplified and sequenced for identification. Overall, 13 biochemical parameters, including proteins, enzymes, lipids, chemicals, and minerals, were measured in serum blood samples from both diseased and healthy fish. The results indicate significant alterations in the levels of these parameters during the outbreak, highlighting the impact of infections on the blood profile of farmed rainbow trout. Urea levels were significantly higher in diseased fish compared to controls, and creatinine, phosphorus, and magnesium also showed similar trends. Alanine aminotransferase and total protein levels were higher in control fish. Chloride levels differed significantly between groups. Iron levels were higher in controls and lower in diseased fish. No significant differences were found in other parameters. This study reveals significant changes in serum blood parameters of rainbow trout during a lactococcosis outbreak caused by L. garvieae. These changes highlight the potential of these parameters as tools for monitoring health status, stress, and aquaculture management. Continuous monitoring can provide valuable insights into disease severity and overall fish health, aiding in the development of improved management practices. The presented data contribute to understanding the pathophysiology of piscine lactococcosis and developing effective mitigation strategies for farmed rainbow trout.
水产养殖业在全球粮食安全方面发挥着至关重要的作用,但它也面临着传染病带来的巨大挑战。虹鳟鱼乳球菌病是虹鳟鱼水产养殖业面临的重大威胁之一,因为它可能导致死亡、生长率下降以及对其他病原体的易感性增加,从而造成严重的经济损失。它给疾病管理策略带来了挑战,影响了虹鳟鱼养殖的可持续性和盈利能力。目前的研究重点是在由加维埃乳球菌(Lactococcus garvieae)引起的乳球菌病爆发期间养殖虹鳟血清血液参数的变化。采集血液样本进行生化分析,检查鱼体内的寄生虫和细菌,并对细菌菌落的 DNA 进行 PCR 扩增和测序鉴定。在患病鱼和健康鱼的血清血液样本中,共测定了 13 项生化参数,包括蛋白质、酶、脂类、化学物质和矿物质。结果表明,在疫情爆发期间,这些参数的水平发生了重大变化,凸显了感染对养殖虹鳟血液特征的影响。与对照组相比,病鱼的尿素水平明显偏高,肌酐、磷和镁也呈现出类似的趋势。对照组鱼的丙氨酸氨基转移酶和总蛋白水平较高。各组之间的氯化物水平差异很大。对照组的铁含量较高,而病鱼的铁含量较低。其他参数没有发现明显差异。这项研究揭示了虹鳟在由加维氏乳球菌引起的乳球菌病爆发期间血清血液参数的显著变化。这些变化凸显了这些参数作为监测健康状况、压力和水产养殖管理工具的潜力。持续监测可为了解疾病严重程度和鱼类总体健康状况提供有价值的信息,有助于制定更好的管理方法。所提供的数据有助于了解鱼类乳球菌病的病理生理学,并为养殖虹鳟制定有效的缓解策略。
{"title":"Changes in blood serum parameters in farmed rainbow trout (Oncorhynchus mykiss) during a piscine lactococcosis outbreak","authors":"Giuseppe Esposito, Stefania Bergagna, Silvia Colussi, Khalid Shahin, Roberta Rosa, Donatella Volpatti, Caterina Faggio, Camilla Mossotto, Alice Gabetti, Alessandra Maganza, Elena Bozzetta, Marino Prearo, Paolo Pastorino","doi":"10.1111/jfd.13994","DOIUrl":"10.1111/jfd.13994","url":null,"abstract":"<p>The aquaculture sector plays a vital role in global food security, yet it grapples with significant challenges posed by infectious diseases. Piscine lactococcosis is one of the significant threats in rainbow trout aquaculture due to its potential to cause severe economic losses through mortalities, reduced growth rates, and increased susceptibility to other pathogens. It poses challenges in disease management strategies, impacting the sustainability and profitability of rainbow trout farming. The current study focuses on the variations in serum blood parameters of farmed rainbow trout <i>Oncorhynchus mykiss</i> during a lactococcosis outbreak caused by <i>Lactococcus garvieae</i>. Blood samples were collected for biochemical analysis, fish were examined for parasites and bacteria, and DNA from bacterial colonies was PCR-amplified and sequenced for identification. Overall, 13 biochemical parameters, including proteins, enzymes, lipids, chemicals, and minerals, were measured in serum blood samples from both diseased and healthy fish. The results indicate significant alterations in the levels of these parameters during the outbreak, highlighting the impact of infections on the blood profile of farmed rainbow trout. Urea levels were significantly higher in diseased fish compared to controls, and creatinine, phosphorus, and magnesium also showed similar trends. Alanine aminotransferase and total protein levels were higher in control fish. Chloride levels differed significantly between groups. Iron levels were higher in controls and lower in diseased fish. No significant differences were found in other parameters. This study reveals significant changes in serum blood parameters of rainbow trout during a lactococcosis outbreak caused by <i>L. garvieae</i>. These changes highlight the potential of these parameters as tools for monitoring health status, stress, and aquaculture management. Continuous monitoring can provide valuable insights into disease severity and overall fish health, aiding in the development of improved management practices. The presented data contribute to understanding the pathophysiology of piscine lactococcosis and developing effective mitigation strategies for farmed rainbow trout.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":"47 10","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jfd.13994","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141476733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xinran Shi, Na Sun, Jiannan Liu, Senting Pu, Zhouling Chen, Dong Sheng, Kexin Hao, Xiaodong Li, Shigen Ye
In this study, the “milky disease” model of Eriocheir sinhensis was constructed via intramuscular injection with the pathogenic yeast Metschnikowia bicuspidata. The dynamic pathological changes of E. sinensis after injection were elucidated with two staining methods (haemotoxylin-eosin and alcian blue periodic acid-Schiff) and fluorescence in situ hybridization technology. Anatomical observation revealed three stages of the “milky disease”: no clinical signs (1–4 days after infection), the appearance of signs of disease (5–7 days), and significant liquefaction (10 days). Histological observation also revealed three stages of the disease: yeast diffusion (1–2 days after infection), yeast slow development (3–4 days), and yeast rapid proliferation (5 days). And FISH technique was suitable for the early detection of infection with M. bicuspidata in E. sinensis. We found that M. bicuspidata spread to the whole body of the crab through the haemolymph and developed into fungal septicaemia. These results elucidate the systemic pathological characteristics of “milky disease” in E. sinensis and suggest the pathogenic mechanism of M. bicuspidata.
{"title":"Dynamic pathology and pathogen distribution of the yeast Metschnikowia bicuspidata in the Chinese mitten crab (Eriocheir sinensis)","authors":"Xinran Shi, Na Sun, Jiannan Liu, Senting Pu, Zhouling Chen, Dong Sheng, Kexin Hao, Xiaodong Li, Shigen Ye","doi":"10.1111/jfd.13993","DOIUrl":"10.1111/jfd.13993","url":null,"abstract":"<p>In this study, the “milky disease” model of <i>Eriocheir sinhensis</i> was constructed via intramuscular injection with the pathogenic yeast <i>Metschnikowia bicuspidata</i>. The dynamic pathological changes of <i>E. sinensis</i> after injection were elucidated with two staining methods (haemotoxylin-eosin and alcian blue periodic acid-Schiff) and fluorescence in situ hybridization technology. Anatomical observation revealed three stages of the “milky disease”: no clinical signs (1–4 days after infection), the appearance of signs of disease (5–7 days), and significant liquefaction (10 days). Histological observation also revealed three stages of the disease: yeast diffusion (1–2 days after infection), yeast slow development (3–4 days), and yeast rapid proliferation (5 days). And FISH technique was suitable for the early detection of infection with <i>M. bicuspidata</i> in <i>E. sinensis</i>. We found that <i>M. bicuspidata</i> spread to the whole body of the crab through the haemolymph and developed into fungal septicaemia. These results elucidate the systemic pathological characteristics of “milky disease” in <i>E. sinensis</i> and suggest the pathogenic mechanism of <i>M. bicuspidata</i>.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":"47 10","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141476734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ruben Avendaño-Herrera, Macarena Echeverría-Bugueño, Mauricio Hernández, Pablo Saldivia, Rute Irgang
A strategy for vaccine design involves identifying proteins that could be involved in pathogen–host interactions. The aim of this proteomic study was to determine how iron limitation affects the protein expression of Tenacibaculum dicentrarchi, with a primary focus on virulence factors and proteins associated with iron uptake. The proteomic analysis was carried out using two strains of T. dicentrarchi grown under normal (control) and iron-limited conditions, mimicking the host environment. Our findings revealed differences in the proteins expressed by the type strain CECT 7612T and the Chilean strain TdCh05 of T. dicentrarchi. Nonetheless, both share a common response to iron deprivation, with an increased expression of proteins associated with iron oxidation and reduction metabolism (e.g., SufA, YpmQ, SufD), siderophore transport (e.g., ExbD, TonB-dependent receptor, HbpA), heme compound biosynthesis, and iron transporters under iron limitation. Proteins involved in gliding motility, such as GldL and SprE, were also upregulated in both strains. A negative differential regulation of metabolic proteins, particularly those associated with amino acid biosynthesis, was observed under iron limitation, reflecting the impact of iron availability on bacterial metabolism. Additionally, the TdCh05 strain exhibited unique proteins associated with gliding motility machinery and phage infection control compared to the type strain. These groups of proteins have been identified as virulence factors within the Flavobacteriaceae family, including the genus Tenacibaculum. These results build upon our previous report on iron acquisition mechanisms and could lay the groundwork for future studies aimed at elucidating the role of some of the described proteins in the infectious process of tenacibaculosis, as well as in the development of potential vaccines.
{"title":"Proteomic characterization of Tenacibaculum dicentrarchi under iron limitation reveals an upregulation of proteins related to iron oxidation and reduction metabolism, iron uptake systems and gliding motility","authors":"Ruben Avendaño-Herrera, Macarena Echeverría-Bugueño, Mauricio Hernández, Pablo Saldivia, Rute Irgang","doi":"10.1111/jfd.13984","DOIUrl":"10.1111/jfd.13984","url":null,"abstract":"<p>A strategy for vaccine design involves identifying proteins that could be involved in pathogen–host interactions. The aim of this proteomic study was to determine how iron limitation affects the protein expression of <i>Tenacibaculum dicentrarchi</i>, with a primary focus on virulence factors and proteins associated with iron uptake. The proteomic analysis was carried out using two strains of <i>T. dicentrarchi</i> grown under normal (control) and iron-limited conditions, mimicking the host environment. Our findings revealed differences in the proteins expressed by the type strain CECT 7612<sup>T</sup> and the Chilean strain TdCh05 of <i>T. dicentrarchi</i>. Nonetheless, both share a common response to iron deprivation, with an increased expression of proteins associated with iron oxidation and reduction metabolism (e.g., SufA, YpmQ, SufD), siderophore transport (e.g., ExbD, TonB-dependent receptor, HbpA), heme compound biosynthesis, and iron transporters under iron limitation. Proteins involved in gliding motility, such as GldL and SprE, were also upregulated in both strains. A negative differential regulation of metabolic proteins, particularly those associated with amino acid biosynthesis, was observed under iron limitation, reflecting the impact of iron availability on bacterial metabolism. Additionally, the TdCh05 strain exhibited unique proteins associated with gliding motility machinery and phage infection control compared to the type strain. These groups of proteins have been identified as virulence factors within the <i>Flavobacteriaceae</i> family, including the genus <i>Tenacibaculum</i>. These results build upon our previous report on iron acquisition mechanisms and could lay the groundwork for future studies aimed at elucidating the role of some of the described proteins in the infectious process of tenacibaculosis, as well as in the development of potential vaccines.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":"47 9","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141468513","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Håvard Bjørgen, Malin Brimsholm, Charlotte Finstad Asserson, Kirstin Skaar, Geir Magne Knutsen, Øyvind Oaland, Randi Haldorsen, Per Gunnar Fjelldal, Tom Hansen, Espen Rimstad, Britta Andrea Kleist, Marius Lund-Iversen, Mariusz Pawel Kowalewski, Erling Olaf Koppang
Melanized focal changes (MFCs) in the fillet of farmed Atlantic salmon is a major quality concern. The changes are thought to initially appear as acute red focal changes (RFCs) that progress into chronic MFCs. Recent findings have indicated that hypoxia may be important in their development, possibly leading to necrosis affecting not only myocytes but also adipocytes. Thus, the aim of this study was to investigate possible hypoxic conditions in RFCs and the subsequent inflammatory responses and lesions in the adipose tissue in RFCs and MFCs. A collection of RFCs, MFCs and control muscle samples from several groups of farmed salmon was studied. Using immunohistochemistry, we found induction of the hypoxia-inducible factor 1 pathway in RFCs. Histological investigations of RFCs and MFCs revealed different stages of fat necrosis, including necrotic adipocytes, a myospherulosis-like reaction and the formation of pseudocystic spaces. Accumulations of foamy macrophages were detected in MFCs, indicating degradation and phagocytosis of lipids. Using in situ hybridization, we showed the presence of tyrosinase- and tyrosinase-related protein-1-expressing amelanotic cells in RFCs, which in turn became melanized in MFCs. In conclusion, we propose a sequence of events leading to the formation of MFCs, highlighting the pivotal role of adiposity, hypoxia and fat necrosis.
{"title":"Deciphering the pathogenesis of melanized focal changes in the white skeletal muscle of farmed Atlantic salmon (Salmo salar)","authors":"Håvard Bjørgen, Malin Brimsholm, Charlotte Finstad Asserson, Kirstin Skaar, Geir Magne Knutsen, Øyvind Oaland, Randi Haldorsen, Per Gunnar Fjelldal, Tom Hansen, Espen Rimstad, Britta Andrea Kleist, Marius Lund-Iversen, Mariusz Pawel Kowalewski, Erling Olaf Koppang","doi":"10.1111/jfd.13988","DOIUrl":"10.1111/jfd.13988","url":null,"abstract":"<p>Melanized focal changes (MFCs) in the fillet of farmed Atlantic salmon is a major quality concern. The changes are thought to initially appear as acute red focal changes (RFCs) that progress into chronic MFCs. Recent findings have indicated that hypoxia may be important in their development, possibly leading to necrosis affecting not only myocytes but also adipocytes. Thus, the aim of this study was to investigate possible hypoxic conditions in RFCs and the subsequent inflammatory responses and lesions in the adipose tissue in RFCs and MFCs. A collection of RFCs, MFCs and control muscle samples from several groups of farmed salmon was studied. Using immunohistochemistry, we found induction of the hypoxia-inducible factor 1 pathway in RFCs. Histological investigations of RFCs and MFCs revealed different stages of fat necrosis, including necrotic adipocytes, a myospherulosis-like reaction and the formation of pseudocystic spaces. Accumulations of foamy macrophages were detected in MFCs, indicating degradation and phagocytosis of lipids. Using in situ hybridization, we showed the presence of tyrosinase- and tyrosinase-related protein-1-expressing amelanotic cells in RFCs, which in turn became melanized in MFCs. In conclusion, we propose a sequence of events leading to the formation of MFCs, highlighting the pivotal role of adiposity, hypoxia and fat necrosis.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":"47 10","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jfd.13988","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141468510","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Arunjyothi Jawaji, Inna Khozin Goldberg, Dina Zilberg
Alternatives to conventional chemical treatments for parasitic diseases in fish are needed. Microalgal-sourced fatty acid ethyl esters (FAEEs) have shown an antiparasitic effect against Gyrodactylus turnbulli infection in guppies. Here, we tested a range of commercial FAEEs of various carbon chain lengths and unsaturation levels against two fish parasites. Guppies and barramundi infected with G. turnbulli and Trichodina sp., respectively, were used. The most effective FAEE, after excluding those toxic to fish, was ethyl laurate (12:0). For both parasites, the LD50 was 18.75 μM within 250 min of incubation. Ethyl eicosapentaenoate (20:5n3) was the next most effective FAEE against G. turnbulli, and dihomo-γ-linolenic acid ethyl ester (20:3n6) and ethyl α-linolenate (18:3n3) were the next most effective against Trichodina sp. In addition, FAEEs prepared from the microalga Phaeodactylum tricornutum residue, after fucoxanthin extraction, were examined against Trichodina sp. infection in barramundi for the first time. LD85 and LD100 was achieved at 2.5 and 5 μL mL−1 of the FAEE preparation, respectively. In vivo, immersion of infected barramundi in 1.25 μL mL−1 of this preparation for 24 h reduced infection prevalence from 100% to 53% and was non-toxic to fish.
{"title":"Exploring the use of fatty acid ethyl esters as a potential natural solution for the treatment of fish parasitic diseases","authors":"Arunjyothi Jawaji, Inna Khozin Goldberg, Dina Zilberg","doi":"10.1111/jfd.13991","DOIUrl":"10.1111/jfd.13991","url":null,"abstract":"<p>Alternatives to conventional chemical treatments for parasitic diseases in fish are needed. Microalgal-sourced fatty acid ethyl esters (FAEEs) have shown an antiparasitic effect against <i>Gyrodactylus turnbulli</i> infection in guppies. Here, we tested a range of commercial FAEEs of various carbon chain lengths and unsaturation levels against two fish parasites. Guppies and barramundi infected with <i>G. turnbulli</i> and <i>Trichodina</i> sp., respectively, were used. The most effective FAEE, after excluding those toxic to fish, was ethyl laurate (12:0). For both parasites, the LD50 was 18.75 μM within 250 min of incubation. Ethyl eicosapentaenoate (20:5<i>n</i>3) was the next most effective FAEE against <i>G. turnbulli</i>, and dihomo-γ-linolenic acid ethyl ester (20:3<i>n</i>6) and ethyl α-linolenate (18:3<i>n</i>3) were the next most effective against <i>Trichodina</i> sp. In addition, FAEEs prepared from the microalga <i>Phaeodactylum tricornutum</i> residue, after fucoxanthin extraction, were examined against <i>Trichodina</i> sp. infection in barramundi for the first time. LD85 and LD100 was achieved at 2.5 and 5 μL mL<sup>−1</sup> of the FAEE preparation, respectively. In vivo, immersion of infected barramundi in 1.25 μL mL<sup>−1</sup> of this preparation for 24 h reduced infection prevalence from 100% to 53% and was non-toxic to fish.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":"47 10","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jfd.13991","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141468511","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Dimitrios Chatzakis, Roula Al Belbeisi, Soultana Karagianni, Eustratios Karagiannakidis, George Koumoundouros
<p>The development of morpho-anatomical abnormalities in fish was first figured in the 16th and 17th centuries (1555–1642, Gudger, <span>1936</span>). In his extensive literature review, Dawson (<span>1964</span>) reported 1020 publications on the presence of morpho-anatomical abnormalities (including skeletal, non-skeletal, and pigmentation defects) in wild fish. A few years later, the same author updated this literature list with 213 new publications, 137 of which were published between 1965 and 1970 (Dawson, <span>1971</span>). Since then, reports on the presence of skeletal abnormalities in wild fish populations are getting more frequent. This might be attributed to the effects of anthropogenic pressures (e.g., pollution, eutrophication) on the stocks (Boglione, <span>2020</span>; Diggles, <span>2013</span>; García-Gasca et al., <span>2016</span>; Leone et al., <span>2021</span>; Messaoudi, Deli, et al., <span>2009</span>), to the exponential increase of scientific awareness about this issue, as well as to the increasing amount of research on bone deformities in aquaculture and the significant research being undertaken to understand the causes (e.g., Boglione et al., <span>2013</span>; Ytteborg et al., <span>2012</span>). Generally, the literature on deformed fish is prevailed by records of one to a few individuals occasionally observed in the field (e.g., Grimaldi & Bertoncini, <span>2021</span>; Jawad & Ibrahim, <span>2018a</span>; Yamamoto et al., <span>2013</span>). Existing studies document that these rates may elevate to high levels, varying among the species (e.g., 11% in <i>Labrus bergylta</i> vs. 57% in <i>Cyclopterus lumpus</i>, Fjelldal et al., <span>2021</span>; 9% in <i>Liza aurata</i> vs. 21% in <i>Chelon labrosus</i>, Boglione et al., <span>2006</span>), and the study area (e.g., 29% in polluted vs 3% in unpolluted sites, <i>Aphanius fasciatus</i>, Kessabi, Annabi, et al., <span>2013</span>; Kessabi, Said, & Messaoudi, <span>2013</span>; 12%–16% in polluted vs. 2%–9% in unpolluted estuaries, García-Gasca et al., <span>2016</span>). Studies on reared fish demonstrate that skeletal abnormalities have significant adverse effects on fish form and function (i.e., growth and survival rates, susceptibility to diseases) (reviewed by Boglione et al., <span>2013</span>). Since these effects are expected to be magnified in the wild, skeletal abnormalities may constitute a significant factor of natural mortality and present a decreasing prevalence with fish age (Kessabi et al., <span>2009</span>; Pollock, <span>2015</span>).</p><p>The status of abnormality prevalence in the wild fish of the Mediterranean Sea remains unknown. As a result, there are no reference data for future monitoring programs and the effect of fish abnormalities on stock natural mortality cannot be estimated. Existing studies on skeletal abnormalities in Mediterranean fishes have primarily focused on the use of fish abnormalities as bioindicators to assess t
7%,表 1)包括血椎区的后凸,伴有轴的最前端异常变直、头骨上移(瞻星者)和身体背侧凹陷(图 2d,d′,e)。在 Pterois miles(Bennett,1828 年)身上也观察到了类似的头骨上移和背侧凹陷现象,其尾足前凸(4.0%,表 1)(图 3a,b)。在该标本中,脊柱前凸与前神经中枢 2 和 3(PU2、PU3)融合成一个带有两个血缘突的中枢有关(图 3a′、b′)。最后,记录到一个 Serranus scriba(Bennett,1828 年)标本(3.3%,表 1)出现前胸前凸,与头骨上移(追星族)有关(图 3c、d)。在本研究记录的脊椎畸形中,腓骨前凸可能是野生鱼类中最常见的报告(例如,Bengtsson 等人,1985 年;Bensaada 等人,2023 年;Jawad 等人,2017 年;Jawad & Ibrahim, 2018a;Mariasingarayan 等人,2022 年;Van Den Avyle 等人,1989 年)。另一方面,据我们所知,这是首次在自然鱼类种群中记录到尾柄严重前凸以及轴的最前端异常变直的情况。在本研究中,记录到的头骨异常包括乙状体区域和上颚受压(大头症)、下颚变短、前下颌缺失以及上下颚侧向错位(交叉咬合)。在一个 Diplodus sargus(Linnaeus,1758 年)样本中观察到的大头症频率最高(28.6%,表 1,图 4a-c)。Pagellus erythrinus (Linnaeus, 1758) 的两个受检样本中都出现了大头鱼和下颌变短的现象(图 4d-g),出现频率相同(每种异常类型的出现率为 1.8%-2.8%,表 1)。在一个红腹角雉标本中,鼻头畸形伴有交叉咬合(图 4e,e′)。在一个鲣鸟标本中,发现了前颌骨缺失的现象(图 4h)。在本研究记录的头骨畸形中,"大头 "是野生鱼类中最常见的畸形(Bueno等人,2015年;Jawad等人,2017年;Näslund & Jawad,2022年;Porta & Snow,2019年)。据我们所知,在饲养的鱼类中经常会出现交叉咬合和下颌缩短的情况(Boglione等人,2013年;Fragkoulis等人,2018年),但在野生鱼类中却很少见。在本研究中,记录到的鱼鳍畸形主要包括背鳍部分或完全缺失,和/或身体背侧凹陷(鞍背综合征)(4个物种,表1)。在 Siganus rivulatus Forsskål & Niebuhr (1775) 的一个标本中,身体背侧的凹陷与鳍条缺失无关,而是与邻近椎骨的后凸和异常形状的神经突有关(图 5a,a′)。在发现的第二个异常的 S. rivulatus 标本和一个 S. cretense 标本中,体背凹陷与邻近的翼管异常和鳐鱼鳍缺失有关(图 5b,b′,c,c′)。一个 D. sargus 标本的体背前部凹陷,解剖学上与背鳍的形状和位置异常有关(图 5d,d″)。最后,Symphodus tinca(Linnaeus,1758 年)和 Solea solea(Linnaeus,1758 年)的单个标本分别出现了背鳍部分全部缺失(图 5e,e′)和尾鳍全部缺失(图 5f,f′)的情况。鞍背综合征(SBS)一词最初由 Tave 等人(1983 年)提出,用于描述罗非鱼 Sarotherodon aureus 背鳍部分至完全缺失的特征。自诞生以来,SBS 已被观察到在人工饲养条件下的各种鱼类(Boglione 等人,2003 年;Cobcroft & Battaglene, 2013 年;Fragkoulis 等人,2017 年;Koumoundouros 等人,2001 年)和全球野生鱼类种群中表现出来(例如,Browder 等人在 10 个鱼类物种中观察到 SBS)、Browder 等人,1993;Koumoundouros,2008;Diggles,2013;Pollock,2015;Jawad & Ibrahim,2018b;Silambarasan 等人,2021;Abed 等人,2023)。值得注意的是,根据 Diggles(2013 年)的记录,在 20 年的时间里,野生 A. australis 的 SBS 患病率增加了约六倍。S. tinca(本研究)的背轮廓缺失部分与 P. miles 报告的异常相似,后者归因于鱼叉捕鱼失败造成的身体伤害(Jimenez 等人,2022 年)、鱼类骨骼异常可能由不利的环境和营养条件(由 Boglione 等人综述,2013 年)、遗传因素(如 Fragkoulis 等人,2020 年;Takeuchi,1966 年;Tave 等人,1983 年)、疾病(Ngo 等人,2024 年;Pasnik 等人,2007 年;Piamsomboon 等人,2022 年)、污染物(如 Johnson 等人,2020 年;Kessabi、Annabi 等人,2000 年)和其他因素引起。
{"title":"Prevalence and typology of skeletal abnormalities in fishes of the Eastern Mediterranean","authors":"Dimitrios Chatzakis, Roula Al Belbeisi, Soultana Karagianni, Eustratios Karagiannakidis, George Koumoundouros","doi":"10.1111/jfd.13992","DOIUrl":"10.1111/jfd.13992","url":null,"abstract":"<p>The development of morpho-anatomical abnormalities in fish was first figured in the 16th and 17th centuries (1555–1642, Gudger, <span>1936</span>). In his extensive literature review, Dawson (<span>1964</span>) reported 1020 publications on the presence of morpho-anatomical abnormalities (including skeletal, non-skeletal, and pigmentation defects) in wild fish. A few years later, the same author updated this literature list with 213 new publications, 137 of which were published between 1965 and 1970 (Dawson, <span>1971</span>). Since then, reports on the presence of skeletal abnormalities in wild fish populations are getting more frequent. This might be attributed to the effects of anthropogenic pressures (e.g., pollution, eutrophication) on the stocks (Boglione, <span>2020</span>; Diggles, <span>2013</span>; García-Gasca et al., <span>2016</span>; Leone et al., <span>2021</span>; Messaoudi, Deli, et al., <span>2009</span>), to the exponential increase of scientific awareness about this issue, as well as to the increasing amount of research on bone deformities in aquaculture and the significant research being undertaken to understand the causes (e.g., Boglione et al., <span>2013</span>; Ytteborg et al., <span>2012</span>). Generally, the literature on deformed fish is prevailed by records of one to a few individuals occasionally observed in the field (e.g., Grimaldi & Bertoncini, <span>2021</span>; Jawad & Ibrahim, <span>2018a</span>; Yamamoto et al., <span>2013</span>). Existing studies document that these rates may elevate to high levels, varying among the species (e.g., 11% in <i>Labrus bergylta</i> vs. 57% in <i>Cyclopterus lumpus</i>, Fjelldal et al., <span>2021</span>; 9% in <i>Liza aurata</i> vs. 21% in <i>Chelon labrosus</i>, Boglione et al., <span>2006</span>), and the study area (e.g., 29% in polluted vs 3% in unpolluted sites, <i>Aphanius fasciatus</i>, Kessabi, Annabi, et al., <span>2013</span>; Kessabi, Said, & Messaoudi, <span>2013</span>; 12%–16% in polluted vs. 2%–9% in unpolluted estuaries, García-Gasca et al., <span>2016</span>). Studies on reared fish demonstrate that skeletal abnormalities have significant adverse effects on fish form and function (i.e., growth and survival rates, susceptibility to diseases) (reviewed by Boglione et al., <span>2013</span>). Since these effects are expected to be magnified in the wild, skeletal abnormalities may constitute a significant factor of natural mortality and present a decreasing prevalence with fish age (Kessabi et al., <span>2009</span>; Pollock, <span>2015</span>).</p><p>The status of abnormality prevalence in the wild fish of the Mediterranean Sea remains unknown. As a result, there are no reference data for future monitoring programs and the effect of fish abnormalities on stock natural mortality cannot be estimated. Existing studies on skeletal abnormalities in Mediterranean fishes have primarily focused on the use of fish abnormalities as bioindicators to assess t","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":"47 10","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-06-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jfd.13992","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141468512","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pacific pink salmon (Oncorhynchus gorbuscha) were deliberately introduced to rivers surrounding the White Sea and has spread to Norway and several other countries surrounding the North Atlantic Ocean. In August 2021, a female pink salmon displaying pale gills and abnormal behaviour was captured in River Lakselva in Northern Norway and later submitted to the Norwegian Veterinary Institute (NVI) for post-mortem examination. Histological examination of organ samples revealed structures indicative of systemic ichthyophoniasis, caused by Ichthyophonus sp. The parasites appeared to be especially abundant in the heart and skeletal musculature, and local tissue responses were assessed to be absent or very mild. Sequences of the ribosomal 18S rRNA and the mitochondrial cytochrome oxidase 1 (CO1) genes confirmed the diagnosis and identified the pathogen as Ichthyophonus sp. The CO1 sequence further established that the isolate from pink salmon was most similar to sequences of Ichthyophonus sp. from Atlantic salmon, Salmo salar, from the Atlantic Ocean on the east coast of the US and from Atlantic herring, Clupea harengus, from Iceland. We here report the first detection of Ichthyophonus sp. in pink salmon in the North Atlantic Ocean.
{"title":"First detection of Ichthyophonus sp. in invasive wild pink salmon (Oncorhynchus gorbuscha) from the North Atlantic Ocean","authors":"Toni Erkinharju, Haakon Hansen, Åse Helen Garseth","doi":"10.1111/jfd.13990","DOIUrl":"10.1111/jfd.13990","url":null,"abstract":"<p>Pacific pink salmon (<i>Oncorhynchus gorbuscha</i>) were deliberately introduced to rivers surrounding the White Sea and has spread to Norway and several other countries surrounding the North Atlantic Ocean. In August 2021, a female pink salmon displaying pale gills and abnormal behaviour was captured in River Lakselva in Northern Norway and later submitted to the Norwegian Veterinary Institute (NVI) for post-mortem examination. Histological examination of organ samples revealed structures indicative of systemic ichthyophoniasis, caused by <i>Ichthyophonus</i> sp. The parasites appeared to be especially abundant in the heart and skeletal musculature, and local tissue responses were assessed to be absent or very mild. Sequences of the ribosomal 18S rRNA and the mitochondrial cytochrome oxidase 1 (CO1) genes confirmed the diagnosis and identified the pathogen as <i>Ichthyophonus</i> sp. The CO1 sequence further established that the isolate from pink salmon was most similar to sequences of <i>Ichthyophonus</i> sp. from Atlantic salmon, <i>Salmo salar</i>, from the Atlantic Ocean on the east coast of the US and from Atlantic herring, <i>Clupea harengus</i>, from Iceland. We here report the first detection of <i>Ichthyophonus</i> sp. in pink salmon in the North Atlantic Ocean.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":"47 9","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jfd.13990","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141457389","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Francisca Samsing, Roisin Sullivan, Ha Truong, Artur Rombenso, Cheryl R. Sangster, Jo Bannister, Matt Longshaw, Joy A. Becker
Fish meal (FM) replacement is essential for the sustainable expansion of aquaculture. This study focussed on the feasibility of replacing FM with a single-cell protein (SCP) derived from methanotrophic bacteria (Methylococcus capsulatus, Bath) in barramundi fry (Lates calcarifer). Three isonitrogenous and isoenergetic diets were formulated with 0%, 6.4% and 12.9% inclusion of the SCP, replacing FM by 0%, 25% and 50%. Barramundi fry (initial body weight 2.5 ± 0.1 g) were fed experimental diets for 21 days to assess growth performance, gut microbiome composition and gut histopathology. Our findings revealed that both levels of SCP inclusion induced detrimental effects in barramundi fry, including impaired growth and reduced survival compared with the control group (66.7% and 71.7% survival in diets replacing FM with SCP by 25% and 50%, respectively; p < .05). Both dietary treatments presented mild necrotizing enteritis with subepithelial oedema and accumulation of PAS positive, diastase resistant droplets within hepatocytes (ceroid hepatopathy) and pancreatic atrophy. Microbiome analysis revealed a marked shift in the gut microbial community with the expansion of potential opportunistic bacteria in the genus Aeromonas. Reduced overall performance in the highest inclusion level (50% SCP) was primarily associated with reduced feed intake, likely related to palatability issues, albeit pathological changes observed in gut and liver may also play a role. Our study highlights the importance of meticulous optimization of SCP inclusion levels in aquafeed formulations, and the need for species and life-stage specific assessments to ensure the health and welfare of fish in sustainable aquaculture practices.
{"title":"Replacement of fishmeal with a microbial single-cell protein induced enteropathy and poor growth outcomes in barramundi (Lates calcarifer) fry","authors":"Francisca Samsing, Roisin Sullivan, Ha Truong, Artur Rombenso, Cheryl R. Sangster, Jo Bannister, Matt Longshaw, Joy A. Becker","doi":"10.1111/jfd.13985","DOIUrl":"10.1111/jfd.13985","url":null,"abstract":"<p>Fish meal (FM) replacement is essential for the sustainable expansion of aquaculture. This study focussed on the feasibility of replacing FM with a single-cell protein (SCP) derived from methanotrophic bacteria (<i>Methylococcus capsulatus</i>, Bath) in barramundi fry (<i>Lates calcarifer</i>). Three isonitrogenous and isoenergetic diets were formulated with 0%, 6.4% and 12.9% inclusion of the SCP, replacing FM by 0%, 25% and 50%. Barramundi fry (initial body weight 2.5 ± 0.1 g) were fed experimental diets for 21 days to assess growth performance, gut microbiome composition and gut histopathology. Our findings revealed that both levels of SCP inclusion induced detrimental effects in barramundi fry, including impaired growth and reduced survival compared with the control group (66.7% and 71.7% survival in diets replacing FM with SCP by 25% and 50%, respectively; <i>p</i> < .05). Both dietary treatments presented mild necrotizing enteritis with subepithelial oedema and accumulation of PAS positive, diastase resistant droplets within hepatocytes (ceroid hepatopathy) and pancreatic atrophy. Microbiome analysis revealed a marked shift in the gut microbial community with the expansion of potential opportunistic bacteria in the genus <i>Aeromonas</i>. Reduced overall performance in the highest inclusion level (<i>50% SCP</i>) was primarily associated with reduced feed intake, likely related to palatability issues, albeit pathological changes observed in gut and liver may also play a role. Our study highlights the importance of meticulous optimization of SCP inclusion levels in aquafeed formulations, and the need for species and life-stage specific assessments to ensure the health and welfare of fish in sustainable aquaculture practices.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":"47 9","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jfd.13985","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141457390","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Edwardsiella anguillarum, a highly virulent species within the Edwardsiella genus, causes significant mortality in milkfish farms in Taiwan. This study aimed to investigate the comparison of milkfish susceptibility, a newly identified host species in Taiwanese aquaculture, with other species Nile tilapia (Oreochromis niloticus) and Asian seabass (Lates calcarifer), to E. anguillarum, elucidating its pathogenicity across both seawater and freshwater aquaculture environments. The results showed milkfish exhibited the highest mortality rate of 85% within 48 h of infection, whereas Nile tilapia exhibited a mortality rate of 70% between the second- and tenth-day post challenge, and seabass exhibited a mortality rate of 25% between the second- and sixth-day post challenge. Gross lesions observed in milkfish included splenomegaly and haemorrhage, whereas Nile tilapia exhibited signs of ascites, exophthalmia and brain haemorrhage. Seabass displayed spleen granulomas and haemorrhage at the injection site. Histopathological analysis revealed common features across all three species, including multifocal necrosis, bacterial presence in the necrotic areas, serositis and oedema. Asian seabass also exhibited chronic lesions in the form of splenic granulomas. This study highlights the high susceptibility of milkfish and Nile tilapia to E. anguillarum, emphasizing the urgent need for further investigation into targeted vaccine development for these fish species. These results not only deepen our understanding of the differing levels of pathogenicity among the three species but also offer valuable insights for improving disease prevention and management strategies in aquaculture, including those applied within polyculture systems and for the maintenance of aquaculture water environments.
{"title":"Comparative pathogenicity and histopathological analysis of Edwardsiella anguillarum intraperitoneal infection in milkfish (Chanos chanos), Nile tilapia (Oreochromis niloticus) and Asian seabass (Lates calcarifer)","authors":"Atiek Rahmawaty, Li-Wu Cheng, Pei-Chi Wang, Shih-Chu Chen","doi":"10.1111/jfd.13982","DOIUrl":"10.1111/jfd.13982","url":null,"abstract":"<p><i>Edwardsiella anguillarum</i>, a highly virulent species within the <i>Edwardsiella</i> genus, causes significant mortality in milkfish farms in Taiwan. This study aimed to investigate the comparison of milkfish susceptibility, a newly identified host species in Taiwanese aquaculture, with other species Nile tilapia (<i>Oreochromis niloticus</i>) and Asian seabass (<i>Lates calcarifer</i>), to <i>E. anguillarum</i>, elucidating its pathogenicity across both seawater and freshwater aquaculture environments. The results showed milkfish exhibited the highest mortality rate of 85% within 48 h of infection, whereas Nile tilapia exhibited a mortality rate of 70% between the second- and tenth-day post challenge, and seabass exhibited a mortality rate of 25% between the second- and sixth-day post challenge. Gross lesions observed in milkfish included splenomegaly and haemorrhage, whereas Nile tilapia exhibited signs of ascites, exophthalmia and brain haemorrhage. Seabass displayed spleen granulomas and haemorrhage at the injection site. Histopathological analysis revealed common features across all three species, including multifocal necrosis, bacterial presence in the necrotic areas, serositis and oedema. Asian seabass also exhibited chronic lesions in the form of splenic granulomas. This study highlights the high susceptibility of milkfish and Nile tilapia to <i>E. anguillarum</i>, emphasizing the urgent need for further investigation into targeted vaccine development for these fish species. These results not only deepen our understanding of the differing levels of pathogenicity among the three species but also offer valuable insights for improving disease prevention and management strategies in aquaculture, including those applied within polyculture systems and for the maintenance of aquaculture water environments.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":"47 9","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141427004","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Aeromonas veronii is an important pathogen found in various aquatic environments and products, posing a threat to public health. The Hanks-like serine/threonine protein kinase is closely linked to the pathogenesis of pathogenic bacteria, but the exact role of YihE in A. veronii remains still unknown. To study the specific function of the YihE kinase, we constructed a knockout mutant of the yihE gene in A. veronii. The deletion of the yihE gene resulted in changes to the metabolism of L-arginine-AMC and acetic acid, as well as enhanced resistance to ampicillin and kanamycin in A. veronii. Additionally, the ΔyihE strain demonstrated a 1.4-fold increase in biofilm formation ability and a 1.8-fold decrease in adhesion and invasion to EPCs when compared to the wild-type strain. A significant decrease in cytotoxicity was observed at 2 and 3 h post-infection with EPCs compared to the wild-type strain. Additionally, the deletion of the yihE gene was associated with a significant decrease in motility of the strain. Furthermore, the deletion of the yihE gene resulted in a 1.44-fold increase in the LD50 of A. veronii in zebrafish. These findings offer valuable insights into the pathogenic mechanisms of A. veronii.
{"title":"The Hanks-like serine/threonine protein kinase YihE is crucial for Aeromonas veronii virulence and adhesion","authors":"Dongjie Shi, Bintong Yang, Tong Zhao, Wenchao Sun, Haiying Cui, Yuanhuan Kang","doi":"10.1111/jfd.13986","DOIUrl":"10.1111/jfd.13986","url":null,"abstract":"<p><i>Aeromonas veronii</i> is an important pathogen found in various aquatic environments and products, posing a threat to public health. The Hanks-like serine/threonine protein kinase is closely linked to the pathogenesis of pathogenic bacteria, but the exact role of YihE in <i>A. veronii</i> remains still unknown. To study the specific function of the YihE kinase, we constructed a knockout mutant of the <i>yihE</i> gene in <i>A. veronii</i>. The deletion of the <i>yihE</i> gene resulted in changes to the metabolism of L-arginine-AMC and acetic acid, as well as enhanced resistance to ampicillin and kanamycin in <i>A. veronii</i>. Additionally, the Δ<i>yihE</i> strain demonstrated a 1.4-fold increase in biofilm formation ability and a 1.8-fold decrease in adhesion and invasion to EPCs when compared to the wild-type strain. A significant decrease in cytotoxicity was observed at 2 and 3 h post-infection with EPCs compared to the wild-type strain. Additionally, the deletion of the <i>yihE</i> gene was associated with a significant decrease in motility of the strain. Furthermore, the deletion of the <i>yihE</i> gene resulted in a 1.44-fold increase in the LD<sub>50</sub> of <i>A. veronii</i> in zebrafish. These findings offer valuable insights into the pathogenic mechanisms of <i>A. veronii</i>.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":"47 9","pages":""},"PeriodicalIF":2.2,"publicationDate":"2024-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141327574","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}