Pub Date : 2025-12-13DOI: 10.1016/j.jip.2025.108517
Simone Roberto Rolando Pisano , Eliane Jemmi , Jonas Steiner, Elodie Cristina, Regula Hirschi, Zoé Delefortrie, Gary Delalay, Heike Schmidt-Posthaus
<div><div><em>Aphanomyces astaci</em>, the etiological agent of crayfish plague, varies in virulence depending on their genetic strains. Five genotype groups (A-E) are well characterized and, with exception of genotype group A, cause significant mortality in European freshwater crayfish species.</div><div>In this study, we aimed to investigate the presence and diversity of <em>A. astaci</em> genotype groups in Switzerland by adapting the qPCR assays (Simplex-qPCR and Duplex-qPCR) published by <span><span>Di Domenico et al. (2021)</span></span> to a Pentaplex-qPCR assay. The Pentaplex-qPCR was compared to Simplex-qPCR, Duplex-qPCR and Triplex-qPCR assays using artificial DNA fragments, culture and tissue samples.</div><div>We performed the Pentaplex-qPCR on DNA extracted from formalin-fixed, paraffin-embedded tissue samples (1991–2020) and fresh tissue samples (2020–2024) of crayfish collected in Switzerland. Crayfish were mainly native crayfish species undergoing a crayfish plague outbreak and few invasive crayfish tested positive for <em>A. astaci</em>.</div><div>The intra-assay repeatability and inter-assay reproducibility of the qPCR was assessed. The Pentaplex-qPCR assay was not inferior to the other qPCR assays (Simplex-qPCR, Duplex-qPCR, Triplex-qPCR) and reliably detected genotype groups A, B, D, and E in 31 of 45 crayfish populations (68.9%) across Switzerland. Most positive samples were linked to crayfish plague outbreaks in European crayfish species, except for four populations, where genotype groups B and D were found in North American signal crayfish (<em>Pacifastacus leniusculus</em>) and Louisiana red swamp crayfish (<em>Procambarus clarkii</em>), respectively. Genotype group C was not detected in any of the samples. Genotype group occurrence and heterogeneity were highest in the Rhine basin, particularly between 2016 and 2020, where groups A, B, D, and E were all detected. In the Ticino basin, genotype group diversity was high and genotype groups B, D and E were detected, while in the Rhone basin, only genotype group B was found. Genotype group D, typically associated with Louisiana red swamp crayfish, was identified in the Rhine basin in 1991 and suspected in 1994, respectively four and one year before the first known detection of its carrier species. Genotype group A was detected once in 2017 in the Rhine basin, causing limited mortality in a European crayfish population.</div><div>The use of archived samples revealed a long history of <em>A. astaci</em> presence, dating back to 1991, providing valuable insights into the spatiotemporal dynamics of crayfish plague. Identifying genotype groups helps clarify potential routes of introduction for both pathogen genotype groups and carrier host species. Furthermore, it facilitates tracking outbreaks and identifying possible primary sources of spread. Future studies are needed to deepen our understanding of the pathogen’s genetic variability in relation to virulence differences and sprea
{"title":"Opening Pandora’s box: First insights into the genetic diversity of Aphanomyces astaci in Switzerland","authors":"Simone Roberto Rolando Pisano , Eliane Jemmi , Jonas Steiner, Elodie Cristina, Regula Hirschi, Zoé Delefortrie, Gary Delalay, Heike Schmidt-Posthaus","doi":"10.1016/j.jip.2025.108517","DOIUrl":"10.1016/j.jip.2025.108517","url":null,"abstract":"<div><div><em>Aphanomyces astaci</em>, the etiological agent of crayfish plague, varies in virulence depending on their genetic strains. Five genotype groups (A-E) are well characterized and, with exception of genotype group A, cause significant mortality in European freshwater crayfish species.</div><div>In this study, we aimed to investigate the presence and diversity of <em>A. astaci</em> genotype groups in Switzerland by adapting the qPCR assays (Simplex-qPCR and Duplex-qPCR) published by <span><span>Di Domenico et al. (2021)</span></span> to a Pentaplex-qPCR assay. The Pentaplex-qPCR was compared to Simplex-qPCR, Duplex-qPCR and Triplex-qPCR assays using artificial DNA fragments, culture and tissue samples.</div><div>We performed the Pentaplex-qPCR on DNA extracted from formalin-fixed, paraffin-embedded tissue samples (1991–2020) and fresh tissue samples (2020–2024) of crayfish collected in Switzerland. Crayfish were mainly native crayfish species undergoing a crayfish plague outbreak and few invasive crayfish tested positive for <em>A. astaci</em>.</div><div>The intra-assay repeatability and inter-assay reproducibility of the qPCR was assessed. The Pentaplex-qPCR assay was not inferior to the other qPCR assays (Simplex-qPCR, Duplex-qPCR, Triplex-qPCR) and reliably detected genotype groups A, B, D, and E in 31 of 45 crayfish populations (68.9%) across Switzerland. Most positive samples were linked to crayfish plague outbreaks in European crayfish species, except for four populations, where genotype groups B and D were found in North American signal crayfish (<em>Pacifastacus leniusculus</em>) and Louisiana red swamp crayfish (<em>Procambarus clarkii</em>), respectively. Genotype group C was not detected in any of the samples. Genotype group occurrence and heterogeneity were highest in the Rhine basin, particularly between 2016 and 2020, where groups A, B, D, and E were all detected. In the Ticino basin, genotype group diversity was high and genotype groups B, D and E were detected, while in the Rhone basin, only genotype group B was found. Genotype group D, typically associated with Louisiana red swamp crayfish, was identified in the Rhine basin in 1991 and suspected in 1994, respectively four and one year before the first known detection of its carrier species. Genotype group A was detected once in 2017 in the Rhine basin, causing limited mortality in a European crayfish population.</div><div>The use of archived samples revealed a long history of <em>A. astaci</em> presence, dating back to 1991, providing valuable insights into the spatiotemporal dynamics of crayfish plague. Identifying genotype groups helps clarify potential routes of introduction for both pathogen genotype groups and carrier host species. Furthermore, it facilitates tracking outbreaks and identifying possible primary sources of spread. Future studies are needed to deepen our understanding of the pathogen’s genetic variability in relation to virulence differences and sprea","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"215 ","pages":"Article 108517"},"PeriodicalIF":2.4,"publicationDate":"2025-12-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145763193","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-10DOI: 10.1016/j.jip.2025.108515
Meifeng Wang , Xiuhua Wang , Jie Huang , Chen Li , Qingli Zhang , Xinyu Lian , Ruoxuan Lu , Hua Xu , Xinyi Fu , Ziyue Gou , Peng Jia , Bing Yang
Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is a widely distributed and highly pathogenic virus affecting shrimp. Helice tientsinensis, a key benthic crab commonly found in shrimp ponds in northern China, has unknown susceptibility to IHHNV and a potential role in viral transmission. In this study, healthy H. tientsinensis individuals were experimentally infected with IHHNV through non-invasive oral way to mimic natural exposure pathways in shrimp aquaculture systems. By day 15 post-infection, some individuals developed small black lesions on the carapace, which mostly resolved after molting. IHHNV-challenged crabs exhibited no mortality. Histopathological examination revealed the presence of Cowdry type A eosinophilic inclusion bodies in hemal sinuses and hepatopancreatic tubules of challenged crabs. In situ hybridization detected strong IHHNV-specific signals in the hepatopancreas, confirming viral infection. Quantitative PCR analysis showed relatively high IHHNV loads in the gills, hepatopancreas, and stomach, with the hepatopancreas exhibiting the highest viral load. Furthermore, this study confirmed cross-species transmission of IHHNV from infected H. tientsinensis to juvenile Penaeus vannamei by qPCR detection and histopathological analysis. These findings meet the WOAH criteria for confirming susceptibility as outlined in Chapter 1.5 of the Aquatic Animal Health Code, supporting the conclusion that H. tientsinensis is a susceptible host of IHHNV. Additionally, a survey conducted from the year 2021 to 2023 revealed the persistent presence of IHHNV in H. tientsinensis populations across various shrimp farming periods along China’s coastal regions, with notably high viral loads detected within pond environments. This study highlights the overlooked risk of cross-species pathogen transmission by wild crab species. The foraging behavior of H. tientsinensis may facilitate the spread of IHHNV within aquaculture systems, posing a potential threat to the health management of shrimp pond farming.
{"title":"A study on the susceptibility of Helice tientsinensis to infectious hypodermal and hematopoietic necrosis virus (IHHNV)","authors":"Meifeng Wang , Xiuhua Wang , Jie Huang , Chen Li , Qingli Zhang , Xinyu Lian , Ruoxuan Lu , Hua Xu , Xinyi Fu , Ziyue Gou , Peng Jia , Bing Yang","doi":"10.1016/j.jip.2025.108515","DOIUrl":"10.1016/j.jip.2025.108515","url":null,"abstract":"<div><div>Infectious hypodermal and hematopoietic necrosis virus (IHHNV) is a widely distributed and highly pathogenic virus affecting shrimp. <em>Helice tientsinensis</em>, a key benthic crab commonly found in shrimp ponds in northern China, has unknown susceptibility to IHHNV and a potential role in viral transmission. In this study, healthy <em>H. tientsinensis</em> individuals were experimentally infected with IHHNV through non-invasive oral way to mimic natural exposure pathways in shrimp aquaculture systems. By day 15 post-infection, some individuals developed small black lesions on the carapace, which mostly resolved after molting. IHHNV-challenged crabs exhibited no mortality. Histopathological examination revealed the presence of Cowdry type A eosinophilic inclusion bodies in hemal sinuses and hepatopancreatic tubules of challenged crabs. <em>In situ</em> hybridization detected strong IHHNV-specific signals in the hepatopancreas, confirming viral infection. Quantitative PCR analysis showed relatively high IHHNV loads in the gills, hepatopancreas, and stomach, with the hepatopancreas exhibiting the highest viral load. Furthermore, this study confirmed cross-species transmission of IHHNV from infected <em>H. tientsinensis</em> to juvenile <em>Penaeus vannamei</em> by qPCR detection and histopathological analysis. These findings meet the WOAH criteria for confirming susceptibility as outlined in Chapter 1.5 of the <em>Aquatic Animal Health Code</em>, supporting the conclusion that <em>H. tientsinensis</em> is a susceptible host of IHHNV. Additionally, a survey conducted from the year 2021 to 2023 revealed the persistent presence of IHHNV in <em>H. tientsinensis</em> populations across various shrimp farming periods along China’s coastal regions, with notably high viral loads detected within pond environments. This study highlights the overlooked risk of cross-species pathogen transmission by wild crab species. The foraging behavior of <em>H. tientsinensis</em> may facilitate the spread of IHHNV within aquaculture systems, posing a potential threat to the health management of shrimp pond farming.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"215 ","pages":"Article 108515"},"PeriodicalIF":2.4,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145734805","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-10DOI: 10.1016/j.jip.2025.108516
Lucas Prado Barreto , Ariel de Souza Oliveira , Dhiogo Neres Carreira , Jaires Gomes de Oliveira Filho , Lorena Lopes Ferreira , Viviane Zeringóta , Lígia Miranda Ferreira Borges , Éverton Kort Kamp Fernandes
The tick Rhipicephalus linnaei (Acari: Ixodidae), formerly recognized as the tropical lineage of Rhipicephalus sanguineus sensu lato (s.l.), is a globally distributed species. Entomopathogenic fungi offer a promising alternative to synthetic acaricides for tick control, yet little is known about the olfactory behavioral response of R. linnaei to fungal exposure. This study used a Y-tube olfactometer to investigate adult ticks’ attraction or repellency to Metarhizium anisopliae conidia and fungal-colonized tick carcasses. Results showed that most adult R. linnaei moved toward the olfactometer arm containing the lowest conidial concentration (1 × 105 conidia mL−1 in mineral oil), indicating attraction. Similarly, most ticks preferred the arm with five fungal-colonized tick carcasses, which resulted from engorged females previously treated with oil conidial suspension. However, higher conidial concentrations or exposure to one or two fungal-colonized carcasses did not elicit attraction or repellency in ticks. This study is the first to report tick behavioral responses to M. anisopliae conidial suspensions and colonized tick carcasses. The observed attraction, although considered reasonably weak, indicates that volatile organic compounds from M. anisopliae IP 119 may affect tick behavior. In addition, oil formulation seems to stimulate the ticks’ behavioral responses to the fungal conidia, potentially by enhancing or suppressing the production of volatile compounds. These findings provide new insights into tick-fungus interactions, supporting the potential use of M. anisopliae in tick management strategies.
{"title":"Behavioral response of the tick Rhipicephalus linnaei to the entomopathogenic fungus Metarhizium anisopliae","authors":"Lucas Prado Barreto , Ariel de Souza Oliveira , Dhiogo Neres Carreira , Jaires Gomes de Oliveira Filho , Lorena Lopes Ferreira , Viviane Zeringóta , Lígia Miranda Ferreira Borges , Éverton Kort Kamp Fernandes","doi":"10.1016/j.jip.2025.108516","DOIUrl":"10.1016/j.jip.2025.108516","url":null,"abstract":"<div><div>The tick <em>Rhipicephalus linnaei</em> (Acari: Ixodidae), formerly recognized as the tropical lineage of <em>Rhipicephalus sanguineus</em> sensu lato (s.l.), is a globally distributed species. Entomopathogenic fungi offer a promising alternative to synthetic acaricides for tick control, yet little is known about the olfactory behavioral response of <em>R. linnaei</em> to fungal exposure. This study used a Y-tube olfactometer to investigate adult ticks’ attraction or repellency to <em>Metarhizium anisopliae</em> conidia and fungal-colonized tick carcasses. Results showed that most adult <em>R. linnaei</em> moved toward the olfactometer arm containing the lowest conidial concentration (1 × 10<sup>5</sup> conidia mL<sup>−1</sup> in mineral oil), indicating attraction. Similarly, most ticks preferred the arm with five fungal-colonized tick carcasses, which resulted from engorged females previously treated with oil conidial suspension. However, higher conidial concentrations or exposure to one or two fungal-colonized carcasses did not elicit attraction or repellency in ticks. This study is the first to report tick behavioral responses to <em>M. anisopliae</em> conidial suspensions and colonized tick carcasses. The observed attraction, although considered reasonably weak, indicates that volatile organic compounds from <em>M. anisopliae</em> IP 119 may affect tick behavior. In addition, oil formulation seems to stimulate the ticks’ behavioral responses to the fungal conidia, potentially by enhancing or suppressing the production of volatile compounds. These findings provide new insights into tick-fungus interactions, supporting the potential use of <em>M. anisopliae</em> in tick management strategies.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"215 ","pages":"Article 108516"},"PeriodicalIF":2.4,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145734804","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-09DOI: 10.1016/j.jip.2025.108514
Alexander M. Gaffke , Jessica L. Griesheimer , Edwin E. Lewis , David Shapiro-Ilan , Fatma Kaplan , Hans T. Alborn
Entomopathogenic nematodes aggregate in the soil environment to facilitate the location and infection of insect hosts. Significant evidence exists suggesting this aggregated movement is controlled, in part, by pheromones produced by individual nematodes. This aggregated movement is often termed trail following. The investigation of pheromones produced by the nematodes at the point of emergence from the insect cadaver, compared to the pheromones produced while moving through substrates, identified differences in pheromone production during these two behaviors. Additionally, the investigation identified differences in pheromone compositions between species, suggesting species specific ratios of the pheromones. Specifically, Steinernema diaprepesi utilized primarily the ascarosides asc-C4 (ascr#11), asc-C5 (ascr#9), asc-C6 (ascr#12), and asc-C7 (ascr#1), while Heterorhabditis bacteriophora primarily utilized the ascarosides asc-C5 (ascr#9), asc-C6 (ascr#12), asc-C11-EA, and asc-C11 (ascr#18). These results indicate that nematodes release different blends of pheromones when dispersing from the cadaver compared to moving through soil, and the two species of nematodes investigated utilized species-specific pheromone compounds. The understanding of nematode behaviors and the chemicals that govern them could prove to be useful in the improvement of biocontrol programs utilizing entomopathogenic nematodes in agricultural settings.
{"title":"Multi-component trail pheromones of the entomopathogenic nematodes Steinernema diaprepesi and Heterorhabditis bacteriophora","authors":"Alexander M. Gaffke , Jessica L. Griesheimer , Edwin E. Lewis , David Shapiro-Ilan , Fatma Kaplan , Hans T. Alborn","doi":"10.1016/j.jip.2025.108514","DOIUrl":"10.1016/j.jip.2025.108514","url":null,"abstract":"<div><div>Entomopathogenic nematodes aggregate in the soil environment to facilitate the location and infection of insect hosts. Significant evidence exists suggesting this aggregated movement is controlled, in part, by pheromones produced by individual nematodes. This aggregated movement is often termed trail following. The investigation of pheromones produced by the nematodes at the point of emergence from the insect cadaver, compared to the pheromones produced while moving through substrates, identified differences in pheromone production during these two behaviors. Additionally, the investigation identified differences in pheromone compositions between species, suggesting species specific ratios of the pheromones. Specifically, Steinernema diaprepesi utilized primarily the ascarosides asc-C4 (ascr#11), asc-C5 (ascr#9), asc-C6 (ascr#12), and asc-C7 (ascr#1), while Heterorhabditis bacteriophora primarily utilized the ascarosides asc-C5 (ascr#9), asc-C6 (ascr#12), asc-C11-EA, and asc-C11 (ascr#18). These results indicate that nematodes release different blends of pheromones when dispersing from the cadaver compared to moving through soil, and the two species of nematodes investigated utilized species-specific pheromone compounds. The understanding of nematode behaviors and the chemicals that govern them could prove to be useful in the improvement of biocontrol programs utilizing entomopathogenic nematodes in agricultural settings.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"215 ","pages":"Article 108514"},"PeriodicalIF":2.4,"publicationDate":"2025-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145734803","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-06DOI: 10.1016/j.jip.2025.108512
Thaís Danielle Santanna , Ethiane Rozo dos Santos , Júlia Jantsch Ferla , Andreza F. de Bem , Bergmann Morais Ribeiro , Daniel M.P. Ardisson-Araújo
Phytophagous mites of the family Tetranychidae cause agricultural losses, yet their viromes remain poorly characterized. We report the first viral survey of the peanut pest mite Tetranychus ogmophallos, identifying three novel positive-sense RNA viruses by high-throughput sequencing and genomic analyses. Tetranychus ogmophallos iflavirus 1 (ToIV-1) possesses a 10,003 nt genome encoding a typical iflavirus polyprotein and shares 58.8% amino acid identity with its closest relative, supporting recognition as a distinct species. Two additional viruses, Tetranychus ogmophallos nodavirus 1 (ToNV-1) and 2 (ToNV-2), were identified as bipartite alphanodaviruses with conserved RNA1:RNA2 abundance ratios of 2–3:1. Strand-specific RT-PCR confirmed genomic and antigenomic strands for ToIV-1 and ToNV-1, indicating active replication. Differential read analysis showed that ToIV-1 was 37-fold more abundant than ToNV-1, suggesting contrasting replication dynamics. These findings expand RNA virus diversity in spider mites and provide a basis for future studies on persistence and potential ecological relevance.
{"title":"Discovery and characterization of RNA viruses in the peanut pest mite Tetranychus ogmophallos (Acari: Tetranychidae)","authors":"Thaís Danielle Santanna , Ethiane Rozo dos Santos , Júlia Jantsch Ferla , Andreza F. de Bem , Bergmann Morais Ribeiro , Daniel M.P. Ardisson-Araújo","doi":"10.1016/j.jip.2025.108512","DOIUrl":"10.1016/j.jip.2025.108512","url":null,"abstract":"<div><div>Phytophagous mites of the family Tetranychidae cause agricultural losses, yet their viromes remain poorly characterized. We report the first viral survey of the peanut pest mite <em>Tetranychus ogmophallos</em>, identifying three novel positive-sense RNA viruses by high-throughput sequencing and genomic analyses. Tetranychus ogmophallos iflavirus 1 (ToIV-1) possesses a 10,003 nt genome encoding a typical iflavirus polyprotein and shares 58.8% amino acid identity with its closest relative, supporting recognition as a distinct species. Two additional viruses, Tetranychus ogmophallos nodavirus 1 (ToNV-1) and 2 (ToNV-2), were identified as bipartite alphanodaviruses with conserved RNA1:RNA2 abundance ratios of 2–3:1. Strand-specific RT-PCR confirmed genomic and antigenomic strands for ToIV-1 and ToNV-1, indicating active replication. Differential read analysis showed that ToIV-1 was 37-fold more abundant than ToNV-1, suggesting contrasting replication dynamics. These findings expand RNA virus diversity in spider mites and provide a basis for future studies on persistence and potential ecological relevance.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"215 ","pages":"Article 108512"},"PeriodicalIF":2.4,"publicationDate":"2025-12-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145708196","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Procambarus clarkii is an economically important cultivated freshwater crayfish species in China. Intensive aquaculture and challenging environmental conditions cause diseases that kill crayfish and impact productivity. Clarification of immune mechanisms could assist the breeding of disease-resistant crayfish and improve survival rate. Here, a tumor necrosis factor alpha-induced protein 8-like gene (TNFAIP8L) encoding a 187 amino acid protein in P. clarkii was characterized, and phylogenetic analysis revealed high homology with genes in other crustaceans. Recombinant TNFAIP8L protein was successfully expressed and purified, and pull-down, mass spectrometry, molecular docking, and western blotting identified hemocyanin B and heat shock protein 60 (HSP60) as TNFAIP8L-interacting proteins. Enzyme-linked immunosorbent assay experiments revealed a higher affinity for hemocyanin B than for HSP60. Expression patterns of TNFAIP8L in different tissues and under immune challenge were determined by real-time PCR. TNFAIP8L was expressed in all tissues examined with highest levels in hemocytes, gills, and intestines. Following immune challenge, TNFAIP8L was down-regulated in hemocytes and gills. RNA interference and overexpression of TNFAIP8L induced the expression of immune-related genes Toll, Serpin, B-cell lymphoma (Bcl), Lectin, Defensin, Crustin, and anti-lipopolysaccharide factor (ALF) in hemocytes and gills. Together, the results suggest that TNFAIP8L mediates immune responses in P. clarkii.
{"title":"Tumor necrosis factor alpha-induced protein 8-like mediates immune responses of Procambarus clarkii through protein–protein interactions","authors":"Gengyu Zhang , Yingying Yu , Xinyue Mei, Cen Qian, Baojian Zhu","doi":"10.1016/j.jip.2025.108502","DOIUrl":"10.1016/j.jip.2025.108502","url":null,"abstract":"<div><div><em>Procambarus clarkii</em> is an economically important cultivated freshwater crayfish species in China. Intensive aquaculture and challenging environmental conditions cause diseases that kill crayfish and impact productivity. Clarification of immune mechanisms could assist the breeding of disease-resistant crayfish and improve survival rate. Here, a tumor necrosis factor alpha-induced protein 8-like gene (<em>TNFAIP8L</em>) encoding a 187 amino acid protein in <em>P. clarkii</em> was characterized, and phylogenetic analysis revealed high homology with genes in other crustaceans. Recombinant TNFAIP8L protein was successfully expressed and purified, and pull-down, mass spectrometry, molecular docking, and western blotting identified hemocyanin B and heat shock protein 60 (HSP60) as TNFAIP8L-interacting proteins. Enzyme-linked immunosorbent assay experiments revealed a higher affinity for hemocyanin B than for HSP60. Expression patterns of <em>TNFAIP8L</em> in different tissues and under immune challenge were determined by real-time PCR. <em>TNFAIP8L</em> was expressed in all tissues examined with highest levels in hemocytes, gills, and intestines. Following immune challenge, <em>TNFAIP8L</em> was down-regulated in hemocytes and gills. RNA interference and overexpression of <em>TNFAIP8L</em> induced the expression of immune-related genes <em>Toll</em>, <em>Serpin</em>, <em>B-cell lymphoma</em> (<em>Bcl</em>), <em>Lectin</em>, <em>Defensin</em>, <em>Crustin</em>, and <em>anti-lipopolysaccharide factor</em> (<em>ALF</em>) in hemocytes and gills. Together, the results suggest that TNFAIP8L mediates immune responses in <em>P. clarkii</em>.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"215 ","pages":"Article 108502"},"PeriodicalIF":2.4,"publicationDate":"2025-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145651843","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-28DOI: 10.1016/j.jip.2025.108503
Feifei Zhu, Tingting Xue, Keping Chen
Glycosylation, a key post-translational protein modification, has been found central to host-pathogen interactions, underpinning numerous critical interactions mediated by carbohydrate structures (glycans). Glycan-dependent interactions regulate pathogen adhesion, recognition, invasion, and immune evasion. Invertebrates represent the largest and most diverse group of species on Earth, from established classical model organisms such as Drosophila and Caenorhabditis, to medically relevant parasites and disease-carrying vectors. However, glycan-mediated interactions between invertebrates and their pathogens remain far less studied compared to vertebrate systems. This review summarizes current knowledge on the role of invertebrate glycosylation, including N- and O-glycans and glycosaminoglycans, in defense against diverse pathogens, and the immune-evasive role of glycosylation employed by invertebrate (or invertebrate-borne) pathogens during host infection. While current research highlights the critical importance of glycosylation in these interactions, some key gaps persist: a lack of comprehensive glycomic and glycoproteomics analysis across representative invertebrate species, and poor understanding of receptor molecules and signaling mechanisms for these glycans and glycoproteins. We hope this review will stimulate further research into this critical yet underexplored facet of invertebrate-pathogen interactions.
{"title":"Glycosylation in invertebrate immunity: Host response and pathogen evasion strategies","authors":"Feifei Zhu, Tingting Xue, Keping Chen","doi":"10.1016/j.jip.2025.108503","DOIUrl":"10.1016/j.jip.2025.108503","url":null,"abstract":"<div><div>Glycosylation, a key post-translational protein modification, has been found central to host-pathogen interactions, underpinning numerous critical interactions mediated by carbohydrate structures (glycans). Glycan-dependent interactions regulate pathogen adhesion, recognition, invasion, and immune evasion. Invertebrates represent the largest and most diverse group of species on Earth, from established classical model organisms such as <em>Drosophila</em> and <em>Caenorhabditis,</em> to medically relevant parasites and disease-carrying vectors. However, glycan-mediated interactions between invertebrates and their pathogens remain far less studied compared to vertebrate systems. This review summarizes current knowledge on the role of invertebrate glycosylation, including N- and O-glycans and glycosaminoglycans, in defense against diverse pathogens, and the immune-evasive role of glycosylation employed by invertebrate (or invertebrate-borne) pathogens during host infection. While current research highlights the critical importance of glycosylation in these interactions, some key gaps persist: a lack of comprehensive glycomic and glycoproteomics analysis across representative invertebrate species, and poor understanding of receptor molecules and signaling mechanisms for these glycans and glycoproteins. We hope this review will stimulate further research into this critical yet underexplored facet of invertebrate-pathogen interactions.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108503"},"PeriodicalIF":2.4,"publicationDate":"2025-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145648729","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Acute hepatopancreatic necrosis disease (AHPN) is a lethal shrimp disease caused by Vibrio parahaemolyticus carrying virulent pirAB genes in extrachromosomal plasmids. However, the limitations of current polymerase chain reaction (PCR) methods pose challenges for on-site AHPND diagnosis. This study aimed to develop a real-time Loop-Mediated Isothermal Amplification (LAMP) assay for diagnosing AHPND at shrimp farming sites. Two primer sets were designed to target a 300 bp region within the pirAB gene on the pVPA3-1 plasmid of the reference strain 13–028/A3. Using a Genie II machine, an AHPND-specific primer set was selected to optimize a LAMP reaction mixture (LAMP-mixture I) and reaction conditions. The LAMP-mixture I provided clear and accurate results at 65 °C within approximately 50 min. The detection limit (100 fg) was comparable to that of duplex PCR developed for AHPND detection and was 100 times more sensitive than conventional PCR with existing LAMP primers (10 pg), though less sensitive than AP4 nested PCR. To enhance the feasibility of the LAMP assay in shrimp farms, a portable real-time LAMP machine developed by SMTION (Daejeon, Korea) was employed. The LAMP products were analyzed using SYBR Green I and calcein detection methods. Both methods produced positive results and showed no cross-reactivity with non-AHPND strains. The real-time LAMP calcein method demonstrated high diagnostic specificity, positive predictive value, and 78 % accuracy when evaluated with field samples. Hence, the real-time LAMP calcein method developed here offers potential for rapid and reliable AHPND diagnosis in shrimp farming sites, in comparison to other PCR-based strategies in simplicity and specificity.
{"title":"Development of a real-time loop-mediated isothermal amplification (real-time LAMP) assay for the onsite detection of Vibrio parahaemolyticus causing acute hepatopancreatic necrosis disease (AHPND) in shrimp","authors":"L.G.T.G. Rajapaksha, C.W.R. Gunasekara, S.H.M.P. Wimalasena, H.N.K.S. Pathirana, Gee-wook Shin","doi":"10.1016/j.jip.2025.108501","DOIUrl":"10.1016/j.jip.2025.108501","url":null,"abstract":"<div><div>Acute hepatopancreatic necrosis disease (AHPN) is a lethal shrimp disease caused by Vibrio parahaemolyticus carrying virulent <em>pir<sup>AB</sup></em> genes in extrachromosomal plasmids. However, the limitations of current polymerase chain reaction (PCR) methods pose challenges for on-site AHPND diagnosis. This study aimed to develop a real-time Loop-Mediated Isothermal Amplification (LAMP) assay for diagnosing AHPND at shrimp farming sites. Two primer sets were designed to target a 300 bp region within the <em>pir<sup>AB</sup></em> gene on the pVPA3-1 plasmid of the reference strain 13–028/A3. Using a Genie II machine, an AHPND-specific primer set was selected to optimize a LAMP reaction mixture (LAMP-mixture I) and reaction conditions. The LAMP-mixture I provided clear and accurate results at 65 °C within approximately 50 min. The detection limit (100 fg) was comparable to that of duplex PCR developed for AHPND detection and was 100 times more sensitive than conventional PCR with existing LAMP primers (10 pg), though less sensitive than AP4 nested PCR. To enhance the feasibility of the LAMP assay in shrimp farms, a portable real-time LAMP machine developed by SMTION (Daejeon, Korea) was employed. The LAMP products were analyzed using SYBR Green I and calcein detection methods. Both methods produced positive results and showed no cross-reactivity with non-AHPND strains. The real-time LAMP calcein method demonstrated high diagnostic specificity, positive predictive value, and 78 % accuracy when evaluated with field samples. Hence, the real-time LAMP calcein method developed here offers potential for rapid and reliable AHPND diagnosis in shrimp farming sites, in comparison to other PCR-based strategies in simplicity and specificity.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108501"},"PeriodicalIF":2.4,"publicationDate":"2025-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145648811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-25DOI: 10.1016/j.jip.2025.108500
Yuanyuan Xu , Jing Zhang , Yan Huang , Dan Zhang , Qiaoling Zhao , Heying Qian , Dongxu Shen
Autophagy is a conserved cellular process with dual roles in antiviral defense and viral utilization that plays a crucial role in host-pathogen interactions. Here, we investigated the function and related mechanisms of Bmced6 in domestic silkworm, Bombyx mori, during the infestation of B. mori nucleopolyhedrovirus (BmNPV). At first, immunofluorescence indicated that Bmced6 was mainly localized in the cytoplasm. And overexpression of Bmced6 significantly enhanced the proliferation of BmNPV, as evidenced by the increased expression level of the viral capsid protein, VP39, and the enhanced viral fluorescence intensity of BmNPV-GFP. Moreover, siRNA-mediated knockdown of Bmced6 inhibited viral infection and transmission. Furthermore, we found that the positive effect of Bmced6 on viral infestation was associated with enhanced virus-induced autophagy , including increased autophagosome formation, ATG8 lipidation, Atg8 puncta formation and up-regulation of autophagy-related genes. Meanwhile, abnormal expression of Bmced6 disrupted mitochondrial homeostasis, leading to ultrastructural damage, decreased mitochondrial membrane potential (MMP), and dysregulated reactive oxygen species (ROS) production. These findings establish that Bmced6 facilitates BmNPV infection through the coupling of autophagy activation and mitochondrial dysfunction, providing new insights into the molecular mechanisms underlying viral infection in insects.
{"title":"Bmced6 enhances BmNPV infection in BmN cells by modulating autophagy and mitochondrial homeostasis","authors":"Yuanyuan Xu , Jing Zhang , Yan Huang , Dan Zhang , Qiaoling Zhao , Heying Qian , Dongxu Shen","doi":"10.1016/j.jip.2025.108500","DOIUrl":"10.1016/j.jip.2025.108500","url":null,"abstract":"<div><div>Autophagy is a conserved cellular process with dual roles in antiviral defense and viral utilization that plays a crucial role in host-pathogen interactions. Here, we investigated the function and related mechanisms of <em>Bmced6</em> in domestic silkworm, <em>Bombyx mori</em>, during the infestation of <em>B. mori</em> nucleopolyhedrovirus (BmNPV). At first, immunofluorescence indicated that Bmced6 was mainly localized in the cytoplasm. And overexpression of <em>Bmced6</em> significantly enhanced the proliferation of BmNPV, as evidenced by the increased expression level of the viral capsid protein, VP39, and the enhanced viral fluorescence intensity of BmNPV-GFP. Moreover, siRNA-mediated knockdown of <em>Bmced6</em> inhibited viral infection and transmission. Furthermore, we found that the positive effect of <em>Bmced6</em> on viral infestation was associated with enhanced virus-induced autophagy , including increased autophagosome formation, ATG8 lipidation, Atg8 puncta formation and up-regulation of autophagy-related genes. Meanwhile, abnormal expression of <em>Bmced6</em> disrupted mitochondrial homeostasis, leading to ultrastructural damage, decreased mitochondrial membrane potential (MMP), and dysregulated reactive oxygen species (ROS) production. These findings establish that <em>Bmced6</em> facilitates BmNPV infection through the coupling of autophagy activation and mitochondrial dysfunction, providing new insights into the molecular mechanisms underlying viral infection in insects.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108500"},"PeriodicalIF":2.4,"publicationDate":"2025-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145620287","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-19DOI: 10.1016/j.jip.2025.108498
Ahlam Ahmed Alfazairy , Yasien Mohamed Gamal El-Abed , Hanan Mohamed Ramadan , Hedaya Hamza Karam , Esmat Mohamed Hegazi
Entomopathogenic protozoans are, to a large extent, host specific, and through acute or chronic infections they negatively alter host reproductive fitness, metabolism, immune response, juvenile hormonal balance, and host development. Hence, these entomopathogens are well-suited to reduce populations of stored product insect pests. The first step towards achieving successful suppression, natural or applied, of storage insect pest populations is the detection for these entomopathogenic protozoans in their habitats. Therefore, a preliminary survey of naturally occurring protozoan infections in stored-grain insect pests was carried out across some Governorates in Lower and Upper Egypt. The protozoan-natural mortality rates among the subject insect pests were recorded. Based on morphological characteristics, particularly spore or oocyst morphology, five entomopathogenic protozoans were taxonomically identified, at the genus level (i.e., four apicomplexans, Adelina sp., Farinocystis sp., Mattesia sp., and Gregarina sp., as well as one microsporidian or fungal pathogen, Nosema sp.). Observations on the morpho-pathological, physio-pathological, and behavioural changes induced by protozoan infections in beetles of Cryptolestes turcicus, Rhyzopertha dominica, Tribolium castaneum, and moths of Plodia interpunctella were recorded, as well. Among the interesting findings, a behavioural abnormality was induced by Nosema infection in P. interpunctella moths; viz., the complete failure of copulated pairs to be separated after copulation (i.e., frequent occurrence of ca., 66.70–73.70 %). Additionally, an increased abundance, by ca., 2.40-fold, of the total protein content has been quantified in Adelina-infected or Farinocystis-infected T. castaneum beetles compared to the uninfected beetles. The pathological changes observed in this study may provide new insights into the interaction between the subject entomopathogenic protozoans and their insect hosts.
{"title":"Observations on morphological, physiological, and behavioural changes induced by protozoan infections in certain storage insect pests","authors":"Ahlam Ahmed Alfazairy , Yasien Mohamed Gamal El-Abed , Hanan Mohamed Ramadan , Hedaya Hamza Karam , Esmat Mohamed Hegazi","doi":"10.1016/j.jip.2025.108498","DOIUrl":"10.1016/j.jip.2025.108498","url":null,"abstract":"<div><div>Entomopathogenic protozoans are, to a large extent, host specific, and through acute or chronic infections they negatively alter host reproductive fitness, metabolism, immune response, juvenile hormonal balance, and host development. Hence, these entomopathogens are well-suited to reduce populations of stored product insect pests. The first step towards achieving successful suppression, natural or applied, of storage insect pest populations is the detection for these entomopathogenic protozoans in their habitats. Therefore, a preliminary survey of naturally occurring protozoan infections in stored-grain insect pests was carried out across some Governorates in Lower and Upper Egypt. The protozoan-natural mortality rates among the subject insect pests were recorded. Based on morphological characteristics, particularly spore or oocyst morphology, five entomopathogenic protozoans were taxonomically identified, at the genus level (i.e.<em>,</em> four apicomplexans, <em>Adelina</em> sp., <em>Farinocystis</em> sp., <em>Mattesia</em> sp., and <em>Gregarina</em> sp., as well as one microsporidian or fungal pathogen, <em>Nosema</em> sp.). Observations on the morpho-pathological, physio-pathological, and behavioural changes induced by protozoan infections in beetles of <em>Cryptolestes turcicus, Rhyzopertha dominica, Tribolium castaneum,</em> and moths of <em>Plodia interpunctella</em> were recorded, as well. Among the interesting findings, a behavioural abnormality was induced by <em>Nosema</em> infection in <em>P. interpunctella</em> moths; viz., the complete failure of copulated pairs to be separated after copulation (i.e., frequent occurrence of ca., 66.70–73.70 %). Additionally, an increased abundance, by <em>ca</em>., 2.40-fold, of the total protein content has been quantified in <em>Adelina</em>-infected or <em>Farinocystis</em>-infected <em>T. castaneum</em> beetles compared to the uninfected beetles. The pathological changes observed in this study may provide new insights into the interaction between the subject entomopathogenic protozoans and their insect hosts.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108498"},"PeriodicalIF":2.4,"publicationDate":"2025-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145573698","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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