Piscirickettsiosis is the main cause of mortality in salmonids of commercial importance in Chile, which is caused by Piscirickettsia salmonis, a Gram-negative, γ-proteobacteria that can produce biofilm as one of its virulence factors. The Chilean salmon industry uses large amounts of antibiotics to control piscirickettsiosis outbreaks, which has raised concern about its environmental impact and the potential to induce antibiotic resistance. Thus, the use of phytogenic feed additives (PFA) with antibacterial activity emerges as an interesting alternative to antimicrobials. Our study describes the antimicrobial action of an Andrographis paniculate-extracted PFA on P. salmonis planktonic growth and biofilm formation. We observed complete inhibition of planktonic and biofilm growth with 500 and 400 μg/mL of PFA for P. salmonis LF-89 and EM-90-like strains, respectively. Furthermore, 500 μg/mL of PFA was bactericidal for both evaluated bacterial strains. Sub-inhibitory doses of PFA increase the transcript levels of stress (groEL), biofilm (pslD), and efflux pump (acrB) genes for both P. salmonis strains in planktonic and sessile conditions. In conclusion, our results demonstrate the antibacterial effect of PFA against P. salmonis in vitro, highlighting the potential of PFA as an alternative to control Piscirickettsiosis.
{"title":"In vitro effects of phytogenic feed additive on Piscirickettsia salmonis growth and biofilm formation","authors":"Natacha Santibáñez, Matías Vega, Tatiana Pérez, Ricardo Enriquez, Carla Estefanía Escalona, Cristian Oliver, Alex Romero","doi":"10.1111/jfd.13913","DOIUrl":"10.1111/jfd.13913","url":null,"abstract":"<p>Piscirickettsiosis is the main cause of mortality in salmonids of commercial importance in Chile, which is caused by <i>Piscirickettsia salmonis</i>, a Gram-negative, γ-proteobacteria that can produce biofilm as one of its virulence factors. The Chilean salmon industry uses large amounts of antibiotics to control piscirickettsiosis outbreaks, which has raised concern about its environmental impact and the potential to induce antibiotic resistance. Thus, the use of phytogenic feed additives (PFA) with antibacterial activity emerges as an interesting alternative to antimicrobials. Our study describes the antimicrobial action of an <i>Andrographis paniculate</i>-extracted PFA on <i>P. salmonis</i> planktonic growth and biofilm formation. We observed complete inhibition of planktonic and biofilm growth with 500 and 400 μg/mL of PFA for <i>P. salmonis</i> LF-89 and EM-90-like strains, respectively. Furthermore, 500 μg/mL of PFA was bactericidal for both evaluated bacterial strains. Sub-inhibitory doses of PFA increase the transcript levels of stress (<i>groEL</i>), biofilm (<i>pslD</i>), and efflux pump (<i>acrB</i>) genes for both <i>P. salmonis</i> strains in planktonic and sessile conditions. In conclusion, our results demonstrate the antibacterial effect of PFA against <i>P. salmonis</i> in vitro, highlighting the potential of PFA as an alternative to control Piscirickettsiosis.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139990207","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}
Carp oedema virus (CEV) has distinct molecularly identified genogroups of viral mutations, denoted as I, IIa, and IIb. Failure to propagate CEV in vitro limits studies towards understanding its interactions with host cells. Here, virus isolates belonging to genogroup I collected during natural outbreaks in the Czech Republic were employed for routine CEV cultivation in monolayers of carp-derived primary cells, common carp brain (CCB) cells, and epithelioma papulosum cyprinid (EPC) cells. Induction of cytopathic effects (CPEs) was observed and recorded in affected cells. Cell survival rate was evaluated under serial dilutions of the CEV inoculum. Virus cell entry was quantified and visualized by qPCR and transmission electron microscopy, respectively. Study findings indicate primary gills epithelia likely present the most suitable matrix for CEV growth in vitro. Cells of the head kidney and spleen facilitate virus entry with microscopically confirmed CPEs and the presence of cytoplasmic pleomorphic virus particles. Cells of the trunk kidney and gonads are unlikely to permit virus cell entry and CPEs development. Although CEV cultivation in cell lines was inconclusive, EPC cells were CEV permissible. Monolayers of carp-derived primary cells show promise for CEV cultivation that could enable elaborate study of mechanisms underlying cellular binding and responses.
{"title":"Potential to grow carp oedema virus (genogroup I) in monolayers of carp-derived primary cells with further implication in cell analysis","authors":"Ehdaa Eltayeb Eltigani Abdelsalam, Zuzana Bláhová, Ali Asghar Baloch, Veronika Piačková","doi":"10.1111/jfd.13934","DOIUrl":"10.1111/jfd.13934","url":null,"abstract":"<p>Carp oedema virus (CEV) has distinct molecularly identified genogroups of viral mutations, denoted as I, IIa, and IIb. Failure to propagate CEV in vitro limits studies towards understanding its interactions with host cells. Here, virus isolates belonging to genogroup I collected during natural outbreaks in the Czech Republic were employed for routine CEV cultivation in monolayers of carp-derived primary cells, common carp brain (CCB) cells, and epithelioma papulosum cyprinid (EPC) cells. Induction of cytopathic effects (CPEs) was observed and recorded in affected cells. Cell survival rate was evaluated under serial dilutions of the CEV inoculum. Virus cell entry was quantified and visualized by qPCR and transmission electron microscopy, respectively. Study findings indicate primary gills epithelia likely present the most suitable matrix for CEV growth in vitro. Cells of the head kidney and spleen facilitate virus entry with microscopically confirmed CPEs and the presence of cytoplasmic pleomorphic virus particles. Cells of the trunk kidney and gonads are unlikely to permit virus cell entry and CPEs development. Although CEV cultivation in cell lines was inconclusive, EPC cells were CEV permissible. Monolayers of carp-derived primary cells show promise for CEV cultivation that could enable elaborate study of mechanisms underlying cellular binding and responses.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jfd.13934","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139990208","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}
Thanh-Nguyen Le, Thuan-Thien Dinh, Thuy-Dung Mai-Hoang, Ebrahim Razzazi-Fazeli, Hieu Tran-Van
Acute Hepatopancreatic Necrosis Disease (AHPND) represents a significant challenge in the field of shrimp aquaculture. This disease is primarily caused by Vibrio parahaemolyticus strains harbouring the pVA1 plasmid encoding the PirAvp and PirBvp toxins. To combat this epidemic and mitigate its devastating consequences, it is crucial to identify and characterize the receptors responsible for the binding of these pathogenic toxins. Our studied discovered that Penaeus vannamei's Serine protease inhibitor 3 (PvSerpin3) derived from shrimp hepatopancreatic tissues could bind to recombinant PirAvp, confirming its role as a novel PirAvp-binding protein (PABP). Through comprehensive computational methods, we revealed two truncated PirAvp–binding proteins derived from PvSerpin3 called Serpin3(13) and Serpin3(22), which had higher affinity to PirAvp than the full-length PvSerpin3. The PABP genes were amplified from a cDNA library that was reversed from total RNA extracted from shrimp, cloned and expressed in Escherichia coli. Three PABP inclusion bodies were refolded to obtain the soluble form, and the recovery efficacy was found to be 100% for Serpin3 and Serpin3(13), while Serpin3(22) had a recovery efficacy of roundly 50%. Co-Immunoprecipitation (co-IP) and dot blot assays substantiated the interaction of these recombinant PABPs with both recombinant PirAvp and VPAHPND (XN89)-producing natural toxins.
{"title":"Serine protease inhibitor 3 (Serpin3) from Penaeus vannamei selectively interacts with Vibrio parahaemolyticus PirAvp","authors":"Thanh-Nguyen Le, Thuan-Thien Dinh, Thuy-Dung Mai-Hoang, Ebrahim Razzazi-Fazeli, Hieu Tran-Van","doi":"10.1111/jfd.13935","DOIUrl":"https://doi.org/10.1111/jfd.13935","url":null,"abstract":"Acute Hepatopancreatic Necrosis Disease (AHPND) represents a significant challenge in the field of shrimp aquaculture. This disease is primarily caused by <i>Vibrio parahaemolyticus</i> strains harbouring the pVA1 plasmid encoding the PirA<sup>vp</sup> and PirB<sup>vp</sup> toxins. To combat this epidemic and mitigate its devastating consequences, it is crucial to identify and characterize the receptors responsible for the binding of these pathogenic toxins. Our studied discovered that <i>Penaeus vannamei'</i>s Serine protease inhibitor 3 (<i>Pv</i>Serpin3) derived from shrimp hepatopancreatic tissues could bind to recombinant PirA<sup>vp</sup>, confirming its role as a novel PirA<sup>vp</sup>-binding protein (P<sub>A</sub>BP). Through comprehensive computational methods, we revealed two truncated PirA<sup>vp</sup>–binding proteins derived from <i>Pv</i>Serpin3 called Serpin3(13) and Serpin3(22), which had higher affinity to PirA<sup>vp</sup> than the full-length <i>Pv</i>Serpin3. The P<sub>A</sub>BP genes were amplified from a cDNA library that was reversed from total RNA extracted from shrimp, cloned and expressed in <i>Escherichia coli</i>. Three P<sub>A</sub>BP inclusion bodies were refolded to obtain the soluble form, and the recovery efficacy was found to be 100% for Serpin3 and Serpin3(13), while Serpin3(22) had a recovery efficacy of roundly 50%. Co-Immunoprecipitation (co-IP) and dot blot assays substantiated the interaction of these recombinant P<sub>A</sub>BPs with both recombinant PirA<sup>vp</sup> and VP<sub>AHPND</sub> (XN89)-producing natural toxins.","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139967491","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}
Ginevra Brocca, Alessandro Truant, Hana Peckova, Martina Lisnerová, Alberto Perolo, Marialetizia Fioravanti, Ivan Fiala, Gianfranco Gabai, Francesco Quaglio, Andrea Gustinelli
Nodular gill disease (NGD) is an emerging condition associated with amoeba trophozoites in freshwater salmonid farms. However, unambiguous identification of the pathogens still must be achieved. This study aimed to identify the amoeba species involved in periodic NGD outbreaks in two rainbow trout (Oncorhynchus mykiss) farms in Northeastern Italy. During four episodes (February–April 2023), 88 fish were euthanized, and their gills were evaluated by macroscopic, microscopic and histopathological examination. The macroscopic and microscopic severity of the lesions and the degree of amoebae infestation were scored and statistically evaluated. One gill arch from each animal was put on non-nutrient agar (NNA) Petri dishes for amoeba isolation, cultivation and subsequent identification with SSU rDNA sequencing. Histopathology confirmed moderate to severe lesions consistent with NGD and mild to moderate amoeba infestation. The presence of amoebae was significantly correlated with lesion severity. Light microscopy of cultured amoebae strains and SSU rDNA analysis revealed the presence of a previously characterized amoeba Naegleria sp. strain GERK and several new strains: two strains from Hartmannelidae, three vannelid amoebae from the genus Ripella and cercozoan amoeba Rosculus. Despite the uncertainty in NGD etiopathogenesis and amoebae pathogenic role, identifying known and new amoebae leans towards a possible multi-aetiological origin.
{"title":"Identification of new amoebae strains in rainbow trout (Oncorhynchus mykiss, Walbaum) farms affected by nodular gill disease (NGD) in Northeastern Italy","authors":"Ginevra Brocca, Alessandro Truant, Hana Peckova, Martina Lisnerová, Alberto Perolo, Marialetizia Fioravanti, Ivan Fiala, Gianfranco Gabai, Francesco Quaglio, Andrea Gustinelli","doi":"10.1111/jfd.13933","DOIUrl":"10.1111/jfd.13933","url":null,"abstract":"<p>Nodular gill disease (NGD) is an emerging condition associated with amoeba trophozoites in freshwater salmonid farms. However, unambiguous identification of the pathogens still must be achieved. This study aimed to identify the amoeba species involved in periodic NGD outbreaks in two rainbow trout (<i>Oncorhynchus mykiss</i>) farms in Northeastern Italy. During four episodes (February–April 2023), 88 fish were euthanized, and their gills were evaluated by macroscopic, microscopic and histopathological examination. The macroscopic and microscopic severity of the lesions and the degree of amoebae infestation were scored and statistically evaluated. One gill arch from each animal was put on non-nutrient agar (NNA) Petri dishes for amoeba isolation, cultivation and subsequent identification with SSU rDNA sequencing. Histopathology confirmed moderate to severe lesions consistent with NGD and mild to moderate amoeba infestation. The presence of amoebae was significantly correlated with lesion severity. Light microscopy of cultured amoebae strains and SSU rDNA analysis revealed the presence of a previously characterized amoeba <i>Naegleria</i> sp. strain GERK and several new strains: two strains from Hartmannelidae, three vannelid amoebae from the genus <i>Ripella</i> and cercozoan amoeba <i>Rosculus</i>. Despite the uncertainty in NGD etiopathogenesis and amoebae pathogenic role, identifying known and new amoebae leans towards a possible multi-aetiological origin.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/jfd.13933","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139940053","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}
Widanarni Widanarni, Muhamad Gustilatov, Julie Ekasari, Pande Gde Sasmita Julyantoro, Diana Elizabeth Waturangi, Sukenda Sukenda
This study aimed to evaluate and unveil the positive impact of biofloc culture on Vibrio parahaemolyticus infection of Pacific white shrimp by reducing quorum sensing (QS) and virulence gene expression and enhancing shrimp's immunity. The shrimp with an average body weight of 0.50 ± 0.09 g were reared in containers with a volume of 2.5 L, 21 units, and a density of 20 shrimp L−1. The shrimp were cultured for 5 days, with each treatment including biofloc system maintenance with a C/N ratio of 10 and a control treatment without biofloc, followed by a challenge test through immersion using V. parahaemolyticus at densities of 103, 105, and 107 CFU mL−1 initially. The results of the in vitro experiment showed that biofloc suspension can inhibit and disperse biofilm formation, as well as reduce the exo-enzyme activity (amylase, protease, and chitinase) of V. parahaemolyticus. Furthermore, the biofloc treatment significantly reduced the expression of the QS regulatory gene OpaR, the PirB toxin gene, and the virulence factor genes T6SS1 and T6SS2 in both in vitro and in vivo. The biofloc system also increased the expression of shrimp immunity-related genes (LGBP, proPO, SP, and PE) and the survival rate of white shrimp challenged with V. parahaemolyticus.
{"title":"Unveiling the positive impact of biofloc culture on Vibrio parahaemolyticus infection of Pacific white shrimp by reducing quorum sensing and virulence gene expression and enhancing immunity","authors":"Widanarni Widanarni, Muhamad Gustilatov, Julie Ekasari, Pande Gde Sasmita Julyantoro, Diana Elizabeth Waturangi, Sukenda Sukenda","doi":"10.1111/jfd.13932","DOIUrl":"10.1111/jfd.13932","url":null,"abstract":"<p>This study aimed to evaluate and unveil the positive impact of biofloc culture on <i>Vibrio parahaemolyticus</i> infection of Pacific white shrimp by reducing quorum sensing (QS) and virulence gene expression and enhancing shrimp's immunity. The shrimp with an average body weight of 0.50 ± 0.09 g were reared in containers with a volume of 2.5 L, 21 units, and a density of 20 shrimp L<sup>−1</sup>. The shrimp were cultured for 5 days, with each treatment including biofloc system maintenance with a C/N ratio of 10 and a control treatment without biofloc, followed by a challenge test through immersion using <i>V</i>. <i>parahaemolyticus</i> at densities of 10<sup>3</sup>, 10<sup>5</sup>, and 10<sup>7</sup> CFU mL<sup>−1</sup> initially. The results of the in vitro experiment showed that biofloc suspension can inhibit and disperse biofilm formation, as well as reduce the exo-enzyme activity (amylase, protease, and chitinase) of <i>V</i>. <i>parahaemolyticus</i>. Furthermore, the biofloc treatment significantly reduced the expression of the QS regulatory gene OpaR, the PirB toxin gene, and the virulence factor genes T6SS1 and T6SS2 in both in vitro and in vivo. The biofloc system also increased the expression of shrimp immunity-related genes (LGBP, proPO, SP, and PE) and the survival rate of white shrimp challenged with <i>V</i>. <i>parahaemolyticus</i>.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139905766","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}
Qijuan Wan, Shaowei Zhai, Minxia Chen, Ming Xu, Songlin Guo
Vibrio harveyi is commonly found in salt and brackish water and is recognized as a serious bacterial pathogen in aquaculture worldwide. In this study, we cloned the ferric uptake regulator (fur) gene from V. harveyi wild-type strain HA_1, which was isolated from diseased American eels (Anguilla rostrata) and has a length of 450 bp, encoding 149 amino acids. Then, a mutant strain, HA_1-Δfur, was constructed through homologous recombination of a suicide plasmid (pCVD442). The HA_1-Δfur mutant exhibited weaker biofilm formation and swarming motility, and 18-fold decrease (5.5%) in virulence to the American eels; compared to the wild-type strain, the mutant strain showed time and diameter differences in growth and haemolysis, respectively. Additionally, the adhesion ability of the mutant strain was significantly decreased. Moreover, there were 15 different biochemical indicators observed between the two strains. Transcriptome analysis revealed that 875 genes were differentially expressed in the Δfur mutant, with 385 up-regulated and 490 down-regulated DEGs. GO and KEGG enrichment analysis revealed that, compared to the wild-type strain, the type II and type VI secretion systems (T2SS and T6SS), amino acid synthesis and transport and energy metabolism pathways were significantly down-regulated, but the ABC transporters and biosynthesis of siderophore group non-ribosomal peptides pathways were up-regulated in the Δfur strain. The qRT-PCR results further confirmed that DEGs responsible for amino acid transport and energy metabolism were positively regulated, but DEGs involved in iron acquisition were negatively regulated in the Δfur strain. These findings suggest that the virulence of the Δfur strain was significantly decreased, which is closely related to phenotype changing and gene transcript regulation.
{"title":"Comparative phenotype and transcriptome analysis revealed the role of ferric uptake regulator (Fur) in the virulence of Vibrio harveyi isolated from diseased American eel (Anguilla rostrata)","authors":"Qijuan Wan, Shaowei Zhai, Minxia Chen, Ming Xu, Songlin Guo","doi":"10.1111/jfd.13931","DOIUrl":"10.1111/jfd.13931","url":null,"abstract":"<p><i>Vibrio harveyi</i> is commonly found in salt and brackish water and is recognized as a serious bacterial pathogen in aquaculture worldwide. In this study, we cloned the ferric uptake regulator (<i>fur</i>) gene from <i>V. harveyi</i> wild-type strain HA_1, which was isolated from diseased American eels (<i>Anguilla rostrata</i>) and has a length of 450 bp, encoding 149 amino acids. Then, a mutant strain, HA_1-Δ<i>fur</i>, was constructed through homologous recombination of a suicide plasmid (pCVD442). The HA_1-Δ<i>fur</i> mutant exhibited weaker biofilm formation and swarming motility, and 18-fold decrease (5.5%) in virulence to the American eels; compared to the wild-type strain, the mutant strain showed time and diameter differences in growth and haemolysis, respectively. Additionally, the adhesion ability of the mutant strain was significantly decreased. Moreover, there were 15 different biochemical indicators observed between the two strains. Transcriptome analysis revealed that 875 genes were differentially expressed in the Δ<i>fur</i> mutant, with 385 up-regulated and 490 down-regulated DEGs. GO and KEGG enrichment analysis revealed that, compared to the wild-type strain, the type II and type VI secretion systems (T2SS and T6SS), amino acid synthesis and transport and energy metabolism pathways were significantly down-regulated, but the ABC transporters and biosynthesis of siderophore group non-ribosomal peptides pathways were up-regulated in the Δ<i>fur</i> strain. The qRT-PCR results further confirmed that DEGs responsible for amino acid transport and energy metabolism were positively regulated, but DEGs involved in iron acquisition were negatively regulated in the Δ<i>fur</i> strain. These findings suggest that the virulence of the Δ<i>fur</i> strain was significantly decreased, which is closely related to phenotype changing and gene transcript regulation.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139900012","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}
Large yellow croaker (Larimichthys crocea) is a vital marine-cultured species in China. Large yellow croaker iridovirus (LYCIV) can cause a high mortality rate in L. crocea. Rapid and convenient detection of LYCIV is an urgent demand for diagnosis. In this study, rapid and simple recombinase polymerase amplification (RPA), real-time RPA and RPA combined with lateral flow dipstick (RPA-LFD) methods were developed for the detection of LYCIV based on the conserved sequence of the LYCIV major capsid protein (MCP) gene. With these optimized RPA analyses, LYCIV detection could be completed within 20 min at 40°C. Both RPA and real-time RPA could detect viral DNA as low as 102 copies/μL, while the detection limit of RPA-LFD was 101 copies/μL, and there was no cross-reaction with other aquatic pathogens (KHV, CyHV-2, GCRV-JX01, SVCV, LCDV and LMBV). In practical evaluation of RPA, real-time RPA and RPA-LFD methods, the results showed consistency with the general PCR detection. In short, the developed RPA, real-time RPA and RPA-LFD analyses could be simple, rapid, sensitive and reliable methods for field diagnosis of LYCIV infection and have significant potential in the protection of LYCIV infection.
{"title":"Rapid and sensitive detection of large yellow croaker iridovirus by real-time RPA and RPA-LFD","authors":"Xiaoru Liu, Yong Cao, Jiayin Wang, Suyuheng Cao, Liqun Lu, Yousheng Jiang","doi":"10.1111/jfd.13930","DOIUrl":"10.1111/jfd.13930","url":null,"abstract":"<p>Large yellow croaker (<i>Larimichthys crocea</i>) is a vital marine-cultured species in China. Large yellow croaker iridovirus (LYCIV) can cause a high mortality rate in <i>L. crocea</i>. Rapid and convenient detection of LYCIV is an urgent demand for diagnosis. In this study, rapid and simple recombinase polymerase amplification (RPA), real-time RPA and RPA combined with lateral flow dipstick (RPA-LFD) methods were developed for the detection of LYCIV based on the conserved sequence of the LYCIV major capsid protein (MCP) gene. With these optimized RPA analyses, LYCIV detection could be completed within 20 min at 40°C. Both RPA and real-time RPA could detect viral DNA as low as 10<sup>2</sup> copies/μL, while the detection limit of RPA-LFD was 10<sup>1</sup> copies/μL, and there was no cross-reaction with other aquatic pathogens (KHV, CyHV-2, GCRV-JX01, SVCV, LCDV and LMBV). In practical evaluation of RPA, real-time RPA and RPA-LFD methods, the results showed consistency with the general PCR detection. In short, the developed RPA, real-time RPA and RPA-LFD analyses could be simple, rapid, sensitive and reliable methods for field diagnosis of LYCIV infection and have significant potential in the protection of LYCIV infection.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139729828","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}
Nasmia Nasmia, Novalina Serdiati, Akbar Marzuki Tahya, Muhammad Safir
Vibrio harveyi and Vibrio parahaemolyticus are species of the Vibrio genus that often cause disease and mass mortality in crustaceans. If not handled quickly and appropriately, these diseases can cause considerable losses to farmers. Therefore, it is necessary to find a solution with safe and environmentally friendly disease prevention technology using natural ingredients, among others from plants, namely oil palm. Some parts of oil palm, namely leaves, fronds, fibres and oil palm pulp, which are palm waste, contain antibacterial compounds. This study aimed to assess the antibacterial activity of palm waste extracts, namely pulp, leaves, fronds and fibres using n-hexane, ethyl acetate, chloroform, ethanol and water maceration solvents against pathogenic bacteria V. harveyi and V. parahaemolyticus, and identify active compounds contained in palm waste. The results of the research are expected to produce innovative and sustainable solutions to control diseases in shrimp farming, contribute to the development of a sustainable fishing industry and open up the potential for utilizing palm waste as a value-added resource in the field of aquatic health. The results of observations on antibacterial activity tests and identifying the content of palm waste extract compounds were analysed descriptively displayed in the form of figures, tables and graphs. The results showed that palm waste extracts (pulp, leaves, fronds and fibres) with ethyl acetate and ethanol maceration solvents had very strong antibacterial potential, namely 20.14 ± 0.31 mm–25.52 ± 1.42 mm on V. harveyi bacteria and 20.41 ± 0.55 mm–25.00 ± 0.51 mm on V. parahaemolyticus bacteria. Palm extracts with n-hexane (>20 mm) and chloroform solvents generally have strong category antibacterial potential (10–20 mm), and palm extracts in water solvents have medium category potential (5–10 mm) against V. harveyi and V. parahemolyticus bacteria. The results of phytochemical tests on palm waste extracts with ethyl acetate and ethanol maceration solvents contain bioactive compounds of flavonoids, saponins, polyphenols and alkaloid tannins, steroids and triterpenoids. Palm extracts with n-hexane and chloroform solvents generally contain saponins, alkaloids, steroids and triterpenoids, while palm waste extracts with water solvents contain saponins.
{"title":"Phytochemical analysis and antibacterial activity of palm waste extract against Vibrio harveyi and Vibrio parahaemolyticus","authors":"Nasmia Nasmia, Novalina Serdiati, Akbar Marzuki Tahya, Muhammad Safir","doi":"10.1111/jfd.13924","DOIUrl":"10.1111/jfd.13924","url":null,"abstract":"<p><i>Vibrio harveyi</i> and <i>Vibrio parahaemolyticus</i> are species of the <i>Vibrio</i> genus that often cause disease and mass mortality in crustaceans. If not handled quickly and appropriately, these diseases can cause considerable losses to farmers. Therefore, it is necessary to find a solution with safe and environmentally friendly disease prevention technology using natural ingredients, among others from plants, namely oil palm. Some parts of oil palm, namely leaves, fronds, fibres and oil palm pulp, which are palm waste, contain antibacterial compounds. This study aimed to assess the antibacterial activity of palm waste extracts, namely pulp, leaves, fronds and fibres using <i>n</i>-hexane, ethyl acetate, chloroform, ethanol and water maceration solvents against pathogenic bacteria <i>V. harveyi</i> and <i>V. parahaemolyticus</i>, and identify active compounds contained in palm waste. The results of the research are expected to produce innovative and sustainable solutions to control diseases in shrimp farming, contribute to the development of a sustainable fishing industry and open up the potential for utilizing palm waste as a value-added resource in the field of aquatic health. The results of observations on antibacterial activity tests and identifying the content of palm waste extract compounds were analysed descriptively displayed in the form of figures, tables and graphs. The results showed that palm waste extracts (pulp, leaves, fronds and fibres) with ethyl acetate and ethanol maceration solvents had very strong antibacterial potential, namely 20.14 ± 0.31 mm–25.52 ± 1.42 mm on <i>V. harveyi</i> bacteria and 20.41 ± 0.55 mm–25.00 ± 0.51 mm on <i>V. parahaemolyticus</i> bacteria. Palm extracts with <i>n</i>-hexane (>20 mm) and chloroform solvents generally have strong category antibacterial potential (10–20 mm), and palm extracts in water solvents have medium category potential (5–10 mm) against <i>V. harveyi</i> and <i>V. parahemolyticus</i> bacteria. The results of phytochemical tests on palm waste extracts with ethyl acetate and ethanol maceration solvents contain bioactive compounds of flavonoids, saponins, polyphenols and alkaloid tannins, steroids and triterpenoids. Palm extracts with <i>n</i>-hexane and chloroform solvents generally contain saponins, alkaloids, steroids and triterpenoids, while palm waste extracts with water solvents contain saponins.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139650898","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}
{"title":"Establishment of a real-time PCR for the detection of decapod iridescent virus 1 (DIV1)","authors":"Xiao-Meng Guo, Jing-Yi Xing, Anqi Li, Liang Qiu, Qing-Li Zhang, Jie Huang","doi":"10.1111/jfd.13926","DOIUrl":"10.1111/jfd.13926","url":null,"abstract":"","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139650897","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}
Teratoma is a rare tumour in fish consisting of tissues from more than one germ layer, that may be located in either the gonads or extragonadal sites. Teratomas in many fish species remain poorly understood. In this work, we performed the first histological examinations of extragonadal teratomas in Poecilia wingei and also examined the influence of a large teratoma on male sexual activity. The studied teratomas shared general organizational features, but they also had variations in both external and internal features. In teratomas, the most common and highly differentiated tissues were striated muscle and nervous tissue. Despite the tumour, the male P. wingei exhibited normal mating behaviour and retained the ability for successful copulation. The structural features of extragonadal teratomas in guppy fish indicate a possible connection between these tumours and a failure of conserved processes operating in the embryonic germline.
{"title":"Investigation of extragonadal teratomas in two Poecilia wingei","authors":"Denis V. Prazdnikov, Ekaterina A. Kondakova","doi":"10.1111/jfd.13929","DOIUrl":"10.1111/jfd.13929","url":null,"abstract":"<p>Teratoma is a rare tumour in fish consisting of tissues from more than one germ layer, that may be located in either the gonads or extragonadal sites. Teratomas in many fish species remain poorly understood. In this work, we performed the first histological examinations of extragonadal teratomas in <i>Poecilia wingei</i> and also examined the influence of a large teratoma on male sexual activity. The studied teratomas shared general organizational features, but they also had variations in both external and internal features. In teratomas, the most common and highly differentiated tissues were striated muscle and nervous tissue. Despite the tumour, the male <i>P. wingei</i> exhibited normal mating behaviour and retained the ability for successful copulation. The structural features of extragonadal teratomas in guppy fish indicate a possible connection between these tumours and a failure of conserved processes operating in the embryonic germline.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":null,"pages":null},"PeriodicalIF":2.5,"publicationDate":"2024-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139642305","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}