During a field survey of parasitic ciliates diversity in South Korea, a scuticociliate was found in a water sample collected during scuba diving. At first glance, the species looks similar to members of the genus Paranophrys especially P. magna but they differ mainly in the body size and the number of somatic kineties. Thus, integrative taxonomic approaches (i.e., silver impregnations, scanning electron microscopy, and molecular phylogeny) were applied and confirmed that the isolate is a new species. Detailed comparison between the new species and the three members of the genus Paranophrys revealed that the new species is similar to P. magna in having membranelle 1 slightly shorter than membranelle 2 and the monokinetids are randomly scattered between dikinetids within each somatic kinety. On the other hand, membranelle 1 in both P. marina (type species) and P. thompsoni is about three times longer than membranelle 2 and their ciliary rows invariably consist of dikinetids anteriorly and monokinetids posteriorly. Observations on the feeding behavior and habitat suggest that the new species is facultative histophagous. Phylogenetic analyses based on 18S rRNA gene indicate that the new species and P. magna belong to the newly established genus Anteparanophrys n. gen. i.e., Anteparanophrys koreana n. sp. and Anteparanophrys magna (Borror, 1972) n. comb.
在韩国对寄生纤毛虫的多样性进行实地调查时，在水肺潜水时收集的水样中发现了一只纤毛虫。乍一看，该物种看起来与Paranophrys属的成员相似，特别是P. magna，但它们的主要区别在于身体大小和身体运动的数量。因此，综合分类方法（即银浸渍、扫描电镜和分子系统发育）证实了该分离物是一个新种。对该新种与该属3个成员的详细比较表明，该新种与P. magna相似，其1号膜略短于2号膜，其单动体在每个体动内随机分布在双动体之间。另一方面，滨海假单胞（模式种）和汤普生假单胞的1号膜比2号膜长约3倍，而且它们的纤毛行总是由前双动体和后单动体组成。对食性和栖地的观察表明该新种为兼性组织食性。基于18S rRNA基因的系统发育分析表明，新种和P. magna属于新建立的Anteparanophrys n. gen，即Anteparanophrys koreana n. sp.和Anteparanophrys magna (Borror, 1972) n. comb。
{"title":"Novel contribution to the taxonomy and molecular phylogeny of scuticociliates (Protozoa): Resolving the non-monophyly of the genus Paranophrys Thompson and Berger, 1965 and the establishment of the genus Anteparanophrys n. gen.","authors":"Jeong Hyeon Yeo , Atef Omar , Sang-Hui Lee , Jae-Ho Jung","doi":"10.1016/j.jip.2025.108267","DOIUrl":"10.1016/j.jip.2025.108267","url":null,"abstract":"<div><div>During a field survey of parasitic ciliates diversity in South Korea, a scuticociliate was found in a water sample collected during scuba diving. At first glance, the species looks similar to members of the genus <em>Paranophrys</em> especially <em>P. magna</em> but they differ mainly in the body size and the number of somatic kineties. Thus, integrative taxonomic approaches (i.e., silver impregnations, scanning electron microscopy, and molecular phylogeny) were applied and confirmed that the isolate is a new species. Detailed comparison between the new species and the three members of the genus <em>Paranophrys</em> revealed that the new species is similar to <em>P. magna</em> in having membranelle 1 slightly shorter than membranelle 2 and the monokinetids are randomly scattered between dikinetids within each somatic kinety. On the other hand, membranelle 1 in both <em>P. marina</em> (type species) and <em>P. thompsoni</em> is about three times longer than membranelle 2 and their ciliary rows invariably consist of dikinetids anteriorly and monokinetids posteriorly. Observations on the feeding behavior and habitat suggest that the new species is facultative histophagous. Phylogenetic analyses based on 18S rRNA gene indicate that the new species and <em>P. magna</em> belong to the newly established genus <em>Anteparanophrys</em> n. gen. i.e., <em>Anteparanophrys koreana</em> n. sp. and <em>Anteparanophrys magna</em> (Borror, 1972) n. comb.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"209 ","pages":"Article 108267"},"PeriodicalIF":3.6,"publicationDate":"2025-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142962190","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 : 2024-12-17DOI: 10.1016/j.jip.2024.108266
Xianzhi Meng , Yixiang Yu , Dandan Ma , Mingxin Mu , Quan Sun , Quanlin Liu , Xiaodong Fan , Tian Li , Jie Chen , Guoqing Pan , Zeyang Zhou
The Pacific white shrimp (Penaeus vannamei), one of the world’s most economically important aquatic species, is highly susceptible to Ecytonucleospora hepatopenaei (EHP), a pathogen that infects the hepatopancreas and causes hepatopancreatic microsporidiosis (HPM), leading to stunted growth and substantial economic losses in shrimp farming. Currently, no effective treatments for EHP exist, making rapid on-site detection and preventive measures essential for disease control. While nucleic acid-based detection methods are commonly employed, they require specialized equipment, controlled environments, and trained personnel, which increase costs. To address this limitation, we developed a colloidal gold immunochromatographic assay (GICA) strip for rapid on-site detection of EHP in shrimp farms. Using LC-MS/MS, 15 high-abundance EHP proteins were identified, with EhSWP3 ranked highest and selected as the optimal antigen detection target. Recombinant EhSWP3 was used to immunize mice, resulting in the development of monoclonal antibodies. The optimal capture and labeled antibody combination (1B6, 3A6) was identified and incorporated into the GICA strip. Testing with common shrimp pathogens and various microsporidia samples demonstrated the high specificity of the EHP test strip. The strip exhibited a sensitivity of 1.81 × 103 copies of the EHP-SSU rRNA gene for detecting EHP-infected shrimp and 1 × 104 purified EHP spores, indicating its strong sensitivity in practical applications. To facilitate on-site use, a simple GICA workflow was established using disposable pestles, Buffer A, and Buffer B, enabling detection within 15 min. Testing of 110 shrimp samples revealed a 90.0 % concordance between the GICA strip and qPCR results. This study marks the first development and application of an EHP antigen detection strip, offering a practical tool for rapid, on-site disease monitoring in shrimp farming.
{"title":"Development of a colloidal gold immunochromatographic strip for the rapid on-site detection of Ecytonucleospora hepatopenaei (EHP)","authors":"Xianzhi Meng , Yixiang Yu , Dandan Ma , Mingxin Mu , Quan Sun , Quanlin Liu , Xiaodong Fan , Tian Li , Jie Chen , Guoqing Pan , Zeyang Zhou","doi":"10.1016/j.jip.2024.108266","DOIUrl":"10.1016/j.jip.2024.108266","url":null,"abstract":"<div><div>The Pacific white shrimp (<em>Penaeus vannamei</em>), one of the world’s most economically important aquatic species, is highly susceptible to <em>Ecytonucleospora hepatopenaei</em> (EHP), a pathogen that infects the hepatopancreas and causes hepatopancreatic microsporidiosis (HPM), leading to stunted growth and substantial economic losses in shrimp farming. Currently, no effective treatments for EHP exist, making rapid on-site detection and preventive measures essential for disease control. While nucleic acid-based detection methods are commonly employed, they require specialized equipment, controlled environments, and trained personnel, which increase costs. To address this limitation, we developed a colloidal gold immunochromatographic assay (GICA) strip for rapid on-site detection of EHP in shrimp farms. Using LC-MS/MS, 15 high-abundance EHP proteins were identified, with EhSWP3 ranked highest and selected as the optimal antigen detection target. Recombinant EhSWP3 was used to immunize mice, resulting in the development of monoclonal antibodies. The optimal capture and labeled antibody combination (1B6, 3A6) was identified and incorporated into the GICA strip. Testing with common shrimp pathogens and various microsporidia samples demonstrated the high specificity of the EHP test strip. The strip exhibited a sensitivity of 1.81 × 10<sup>3</sup> copies of the EHP<em>-SSU rRNA</em> gene for detecting EHP-infected shrimp and 1 × 10<sup>4</sup> purified EHP spores, indicating its strong sensitivity in practical applications. To facilitate on-site use, a simple GICA workflow was established using disposable pestles, Buffer A, and Buffer B, enabling detection within 15 min. Testing of 110 shrimp samples revealed a 90.0 % concordance between the GICA strip and qPCR results. This study marks the first development and application of an EHP antigen detection strip, offering a practical tool for rapid, on-site disease monitoring in shrimp farming.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"209 ","pages":"Article 108266"},"PeriodicalIF":3.6,"publicationDate":"2024-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142864585","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 : 2024-12-14DOI: 10.1016/j.jip.2024.108265
Emily D. Vu , Sijun Liu , Bryony C. Bonning
The small hive beetle (SHB), Aethina tumida Murray is an invasive pest of the honey bee. This beetle feeds not only on bee resources within the hive such as honey and pollen, but also on bee brood and dead bees. The impact of this beetle’s intimate parasitic association with the honey bee on virus transmission is poorly understood. We aimed to characterize the virome of SHB to identify SHB viruses with potential for use in biological control of this pest. We characterized the virome of SHB by sequencing the transcriptomes and small RNAs of SHB collected from multiple geographical regions: Adult and larval SHB were collected from midwestern- (Illinois, Ohio) and southern- (Florida, Texas) states of the USA, and from South Africa. Small RNAs were sequenced for adult beetles from Florida and Ohio, for larvae from Florida, and for an SHB-derived cell line (BCIRL-AtumEN-1129). Assembled transcripts were annotated by BLASTx. In field-caught adult beetles and adults and larvae from South Africa, the near-complete sequences for all three genomic segments of a putative novel phasmavirus (order: Elliovirales, formerly Bunyavirales) were identified. In addition, transcripts from a partial glycoprotein sequence from a different phasmavirus integrated into the genome of SHB were detected in all samples, including the SHB-derived cell line. Apparent PIWI-interacting RNAs derived from the integrated glycoprotein sequence were also detected. Whether the putative extant phasmavirus replicates in SHB remains to be determined.
{"title":"Phasmavirus-derived genome sequences and endogenous viral element identified in the small hive beetle, Aethina tumida Murray","authors":"Emily D. Vu , Sijun Liu , Bryony C. Bonning","doi":"10.1016/j.jip.2024.108265","DOIUrl":"10.1016/j.jip.2024.108265","url":null,"abstract":"<div><div>The small hive beetle (SHB), <em>Aethina tumida</em> Murray is an invasive pest of the honey bee. This beetle feeds not only on bee resources within the hive such as honey and pollen, but also on bee brood and dead bees. The impact of this beetle’s intimate parasitic association with the honey bee on virus transmission is poorly understood. We aimed to characterize the virome of SHB to identify SHB viruses with potential for use in biological control of this pest. We characterized the virome of SHB by sequencing the transcriptomes and small RNAs of SHB collected from multiple geographical regions: Adult and larval SHB were collected from midwestern- (Illinois, Ohio) and southern- (Florida, Texas) states of the USA, and from South Africa. Small RNAs were sequenced for adult beetles from Florida and Ohio, for larvae from Florida, and for an SHB-derived cell line (BCIRL-AtumEN-1129). Assembled transcripts were annotated by BLASTx. In field-caught adult beetles and adults and larvae from South Africa, the near-complete sequences for all three genomic segments of a putative novel phasmavirus (order: <em>Elliovirales,</em> formerly <em>Bunyavirales</em>) were identified. In addition, transcripts from a partial glycoprotein sequence from a different phasmavirus integrated into the genome of SHB were detected in all samples, including the SHB-derived cell line. Apparent PIWI-interacting RNAs derived from the integrated glycoprotein sequence were also detected. Whether the putative extant phasmavirus replicates in SHB remains to be determined.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"209 ","pages":"Article 108265"},"PeriodicalIF":3.6,"publicationDate":"2024-12-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142828948","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 : 2024-12-10DOI: 10.1016/j.jip.2024.108258
Alexandria N. Payne , Vincent Prayugo , Adam G. Dolezal
Corpse-mediated pathogen transmission is a viable route through which naïve hosts can become infected, but its likelihood for honey bee-associated viruses is largely unknown. While these viruses can be easily detected in deceased bees, it remains unclear if they stay infectious within postmortem hosts or if enough viral RNA degradation—and subsequently virus inactivation—occurs post-host death to render these viruses inviable. This knowledge gap has important implications for how researchers perform honey bee virus studies and for our general understanding of honey bee virus transmission. To better understand the resiliency of honey bee-associated viruses within deceased hosts, we first tested the hypothesis that postmortem specimens, stored in colony-normal temperature and humidity conditions, can be reliably used to quantify virus abundance. To determine this, we experimentally-infected adult honey bees with Israeli acute paralysis virus (IAPV) and then measured the virus levels of individuals sampled live or at different postmortem time points (4, 12, 24, and 48 hours post-death) using RT-qPCR and a standard curve absolute quantification method. We found no significant differences based on when bees were sampled, indicating that postmortem honey bees are statistically comparable to using live-sampled bees and can be reliably used to quantify absolute IAPV abundance. We then performed a follow-up experiment that determined whether or not the IAPV detected in postmortem bees remained infectious over time. We found that IAPV extracted from postmortem bees remained highly infectious for at least 48 hours post-death, indicating that any viral RNA degradation that may have occurred during the postmortem interval did not adversely affect IAPV’s overall infectivity. The results from this study suggest that IAPV is more resilient to degradation than previously assumed, support the use of postmortem bees for downstream IAPV analyses, and indicate that postmortem hosts can act as sources of IAPV infection for susceptible individuals.
{"title":"A honey bee-associated virus remains infectious and quantifiable in postmortem hosts","authors":"Alexandria N. Payne , Vincent Prayugo , Adam G. Dolezal","doi":"10.1016/j.jip.2024.108258","DOIUrl":"10.1016/j.jip.2024.108258","url":null,"abstract":"<div><div>Corpse-mediated pathogen transmission is a viable route through which naïve hosts can become infected, but its likelihood for honey bee-associated viruses is largely unknown. While these viruses can be easily detected in deceased bees, it remains unclear if they stay infectious within postmortem hosts or if enough viral RNA degradation—and subsequently virus inactivation—occurs post-host death to render these viruses inviable. This knowledge gap has important implications for how researchers perform honey bee virus studies and for our general understanding of honey bee virus transmission. To better understand the resiliency of honey bee-associated viruses within deceased hosts, we first tested the hypothesis that postmortem specimens, stored in colony-normal temperature and humidity conditions, can be reliably used to quantify virus abundance. To determine this, we experimentally-infected adult honey bees with Israeli acute paralysis virus (IAPV) and then measured the virus levels of individuals sampled live or at different postmortem time points (4, 12, 24, and 48 hours post-death) using RT-qPCR and a standard curve absolute quantification method. We found no significant differences based on when bees were sampled, indicating that postmortem honey bees are statistically comparable to using live-sampled bees and can be reliably used to quantify absolute IAPV abundance. We then performed a follow-up experiment that determined whether or not the IAPV detected in postmortem bees remained infectious over time. We found that IAPV extracted from postmortem bees remained highly infectious for at least 48 hours post-death, indicating that any viral RNA degradation that may have occurred during the postmortem interval did not adversely affect IAPV’s overall infectivity. The results from this study suggest that IAPV is more resilient to degradation than previously assumed, support the use of postmortem bees for downstream IAPV analyses, and indicate that postmortem hosts can act as sources of IAPV infection for susceptible individuals.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"209 ","pages":"Article 108258"},"PeriodicalIF":3.6,"publicationDate":"2024-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142818344","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 : 2024-12-05DOI: 10.1016/j.jip.2024.108251
F. Rueda-Maíllo , I. Garrido-Jurado , I. Kotta-Loizou , E. Quesada-Moraga
Entomopathogenic ascomycetes are important natural regulators of insect pest populations and an increasingly adopted microbial control option. Fungal virulence in entomopathogenic ascomycetes can be modified by mycoviruses, viruses that infect fungi, whereas the possible role of these viruses on the physical and biochemical properties of the virus-containing fungal strains and on their ecological fitness has remained largely unexplored. Here, utilizing a Beauveria bassiana strain naturally infected with two mycoviruses, Beauveria bassiana partitivirus 2 (BbPV-2) and Beauveria bassiana polymycovirus 1 (BbPmV-1), we found that the mycovirus-containing strain is hypervirulent towards the experimental insect Galleria mellonella and shows major physical and biochemical changes in spore size, isoelectric point, and Pr1 activity, but even more impactful, the mycoviral infection confers a significant environmental- abiotic and biotic stress tolerance to the fungus. Hence, mycovirus infection expanded the temperature range for fungal growth and germination, and improved tolerance to osmotic stress, water stress, and UV-B radiation. Similarly, the antagonistic activity of the mycovirus-containing strain against Trichoderma harzianum was increased as compared to the mycovirus-free one. Taken together, these data suggest for the first time a mycovirus related adaptation of key traits indicators of environmental competence of a beneficial fungus, rendering these mycoviruses as potent tools for entomopathogenic fungal strain selection and development as mycoinsecticides.
{"title":"A mycoviral infection drives virulence and ecological fitness of the entomopathogenic fungus Beauveria bassiana","authors":"F. Rueda-Maíllo , I. Garrido-Jurado , I. Kotta-Loizou , E. Quesada-Moraga","doi":"10.1016/j.jip.2024.108251","DOIUrl":"10.1016/j.jip.2024.108251","url":null,"abstract":"<div><div>Entomopathogenic ascomycetes are important natural regulators of insect pest populations and an increasingly adopted microbial control option. Fungal virulence in entomopathogenic ascomycetes can be modified by mycoviruses, viruses that infect fungi, whereas the possible role of these viruses on the physical and biochemical properties of the virus-containing fungal strains and on their ecological fitness has remained largely unexplored. Here, utilizing a <em>Beauveria bassiana</em> strain naturally infected with two mycoviruses, Beauveria bassiana partitivirus 2 (BbPV-2) and Beauveria bassiana polymycovirus 1 (BbPmV-1), we found that the mycovirus-containing strain is hypervirulent towards the experimental insect <em>Galleria mellonella</em> and shows major physical and biochemical changes in spore size, isoelectric point, and Pr1 activity, but even more impactful, the mycoviral infection confers a significant environmental- abiotic and biotic stress tolerance to the fungus. Hence, mycovirus infection expanded the temperature range for fungal growth and germination, and improved tolerance to osmotic stress, water stress, and UV-B radiation. Similarly, the antagonistic activity of the mycovirus-containing strain against <em>Trichoderma harzianum</em> was increased as compared to the mycovirus-free one. Taken together, these data suggest for the first time a mycovirus related adaptation of key traits indicators of environmental competence of a beneficial fungus, rendering these mycoviruses as potent tools for entomopathogenic fungal strain selection and development as mycoinsecticides.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"209 ","pages":"Article 108251"},"PeriodicalIF":3.6,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142791702","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-12-05DOI: 10.1016/j.jip.2024.108243
Ralf J. Sommer
The free-living nematode Pristionchus pacificus has been established as a model system in integrative evolutionary biology by combining laboratory studies with field work and evolutionary biology. Multiple genetic, molecular and experimental tools and a collection of more than 2,500 P. pacificus strains and more than 50 Pristionchus species, which are available as living cultures or frozen stock collections, support research on various life history traits. Species of Pristionchus exhibit a number of complex traits unknown from Caenorhabditis elegans and most other free-living nematodes. First, P. pacificus can form two alternative mouth forms, an example of developmental plasticity that is increasingly studied to investigate the role of plasticity as a facilitator of evolutionary novelty. More than a decade of work has identified associated genetic and epigenetic mechanisms and revealed the evolutionary and ecological significance of feeding structure plasticity. Second, one of the two mouth morphs results in predatory behavior against other nematodes and is currently used to investigate the neurobiology of predation. Third, potential predation results in the risk of cannibalism among conspecifics. Strikingly, Pristionchus nematodes have developed a self-recognition system that allows the distinction of self (kin) and non-self. Given all these organismal features, this nematode has recently been considered a key example for research towards a new natural history (West-Eberhard, 2024). Here, I summarize recent work on Pristionchus with a focus on a ‘new natural history’. In addition, I review some recent studies that indicate an interaction of Pristionchus with EPNs that was suggested based on various surveys in different ecological habitats.
{"title":"Pristionchus – Beetle associations: Towards a new natural history","authors":"Ralf J. Sommer","doi":"10.1016/j.jip.2024.108243","DOIUrl":"10.1016/j.jip.2024.108243","url":null,"abstract":"<div><div>The free-living nematode <em>Pristionchus pacificus</em> has been established as a model system in integrative evolutionary biology by combining laboratory studies with field work and evolutionary biology. Multiple genetic, molecular and experimental tools and a collection of more than 2,500 <em>P. pacificus</em> strains and more than 50 <em>Pristionchus</em> species, which are available as living cultures or frozen stock collections, support research on various life history traits. Species of <em>Pristionchus</em> exhibit a number of complex traits unknown from <em>Caenorhabditis elegans</em> and most other free-living nematodes. First, <em>P. pacificus</em> can form two alternative mouth forms, an example of developmental plasticity that is increasingly studied to investigate the role of plasticity as a facilitator of evolutionary novelty. More than a decade of work has identified associated genetic and epigenetic mechanisms and revealed the evolutionary and ecological significance of feeding structure plasticity. Second, one of the two mouth morphs results in predatory behavior against other nematodes and is currently used to investigate the neurobiology of predation. Third, potential predation results in the risk of cannibalism among conspecifics. Strikingly, <em>Pristionchus</em> nematodes have developed a self-recognition system that allows the distinction of self (kin) and non-self. Given all these organismal features, this nematode has recently been considered a key example for research towards a new natural history (<span><span>West-Eberhard, 2024</span></span>). Here, I summarize recent work on <em>Pristionchus</em> with a focus on a ‘new natural history’. In addition, I review some recent studies that indicate an interaction of <em>Pristionchus</em> with EPNs that was suggested based on various surveys in different ecological habitats.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"209 ","pages":"Article 108243"},"PeriodicalIF":3.6,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142791758","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-12-04DOI: 10.1016/j.jip.2024.108238
Lorrane de Andrade Pereira , Sandra Maria de Oliveira Souza , Margareth Maria de Carvalho Queiroz , Rafael Meyer Mariante , Viviane Zahner
Brevibacillus laterosporus is a sporulating bacteria, with typical canoe-shaped parasporal bodies attached to the spores. It has shown great biotechnological potential, including its broad pathogenic spectrum against different orders of insects and other invertebrates with medical-veterinary-sanitary importance. The high degree of synanthropy of muscoid dipterans and the damage they cause being a source of myiasis and carriers of pathogens encourage research into their population control. The objective of the study was to evaluate the histopathological effects caused by B. laterosporus NRS 590 on larvae and adults of M. domestica and C. megacephala. The flies were collected in garbage dumpsters, identified and the respective colonies were adapted to laboratory conditions (air-conditioned chamber with regulated humidity and temperature). Different concentrations of bacterial spore suspensions were offered to neo larvae and adults. Larvae and adults were dissected and intestinal epithelial cells of the flies, in temporal kinetics, were demonstrated using transmission electron microscopy (TEM) techniques. Irregular microvilli were observed in the intestines of both adult and larvae tested after 6 h of treatment. At 12 h and 24 h, intense cellular disorganization was observed and at 24 h and 48 h there was discontinuity of microvilli, cell extrusion and nuclear damage in some cells, as well. Spores of B. laterosporus NRS590 produce histopathological effects on both larvae and adults of synanthropic flies, being an alternative active principle in eco-friendly insecticides.
{"title":"Ultrastructural changes in epithelial cells on different stages of sinantropic muscoid dipterans fed with spores of Brevibacillus laterosporus","authors":"Lorrane de Andrade Pereira , Sandra Maria de Oliveira Souza , Margareth Maria de Carvalho Queiroz , Rafael Meyer Mariante , Viviane Zahner","doi":"10.1016/j.jip.2024.108238","DOIUrl":"10.1016/j.jip.2024.108238","url":null,"abstract":"<div><div><em>Brevibacillus laterosporus</em> is a sporulating bacteria, with typical canoe-shaped parasporal bodies attached to the spores. It has shown great biotechnological potential, including its broad pathogenic spectrum against different orders of insects and other invertebrates with medical-veterinary-sanitary importance. The high degree of synanthropy of muscoid dipterans and the damage they cause being a source of myiasis and carriers of pathogens encourage research into their population control. The objective of the study was to evaluate the histopathological effects caused by <em>B. laterosporus</em> NRS 590 on larvae and adults of <em>M. domestica</em> and <em>C. megacephala</em>. The flies were collected in garbage dumpsters, identified and the respective colonies were adapted to laboratory conditions (air-conditioned chamber with regulated humidity and temperature). Different concentrations of bacterial spore suspensions were offered to neo larvae and adults. Larvae and adults were dissected and intestinal epithelial cells of the flies, in temporal kinetics, were demonstrated using transmission electron microscopy (TEM) techniques. Irregular microvilli were observed in the intestines of both adult and larvae tested after 6 h of treatment. At 12 h and 24 h, intense cellular disorganization was observed and at 24 h and 48 h there was discontinuity of microvilli, cell extrusion and nuclear damage in some cells, as well. Spores of <em>B. laterosporus</em> NRS590 produce histopathological effects on both larvae and adults of synanthropic flies, being an alternative active principle in eco-friendly insecticides.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"209 ","pages":"Article 108238"},"PeriodicalIF":3.6,"publicationDate":"2024-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142791774","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 : 2024-12-04DOI: 10.1016/j.jip.2024.108241
Manoely Abreu Reis , Felipe Marinho Coutinho de Souza , Ianne Caroline da Silva Nobre , Fátima Maryelen Gomes de Fraga Dias , Maria Fátima Grossi-de-Sá , José Dijair Antonino
The humoral response plays a crucial role in insect defense against parasites and pathogens, typically producing antimicrobial peptides through the Toll, IMD, and Jak-STAT signaling pathways, as well as melanization via phenoloxidases. However, many studies use nonpathogenic or opportunistic organisms and often infect insects in nonnatural ways, such as piercing or injecting the pathogen into the hemocoel. The objective of this study was to examine the modulation of the main humoral pathway genes involved in the interaction between the nonmodel organism Diatraea saccharalis (the sugarcane borer) and different biological control agents. We identified and evaluated the expression of DsDorsal (Toll pathway), DsRelish (IMD pathway), DsSTAT (JAK/STAT pathway), DsPPO1, and DsPPO2 (PO pathway) in larvae and pupae of D. saccharalis exposed or not to different biological control agents. The biocontrol agents used were: (i) the bacterium Bacillus thuringiensis var. aizawai GC-91, which is pathogenic to D. saccharalis; (ii) the fungus Metarhizium anisopliae ESALQE9 strain, which is employed to control the froghoppers of the genus Mahanarva in sugarcane fields, though it exhibits low virulence to D. saccharalis; and (iii) the generalist parasitoid Tetrastichus howardi. Our results demonstrate that B. thuringiensis at LC30 induced the expression of DsRelish at 24 h and DsSTAT at 48 h after treatment initiation. In contrast, treatment with the M. anisopliae ESALQE9 strain reduced the levels of DsDorsal and DsSTAT 24 h post-infection compared to the control group. In larvae, DsDorsal, DsSTAT, DsPPO1, and DsPPO2 were induced in response to T. howardi, whereas no induction was observed in pupae. Notably, no immune-related genes were modulated during the pupae-parasitoid interaction. Additionally, we provide an explanation for why T. howardi shows superior parasitism success in D. saccharalis pupae compared with larvae. The data presented here introduce novel perspectives for enhancing pest management through the utilization of biocontrol agents.
{"title":"Distinct biological control agents differentially modulate the immune system of the sugarcane borer larvae (Diatraea saccharalis)","authors":"Manoely Abreu Reis , Felipe Marinho Coutinho de Souza , Ianne Caroline da Silva Nobre , Fátima Maryelen Gomes de Fraga Dias , Maria Fátima Grossi-de-Sá , José Dijair Antonino","doi":"10.1016/j.jip.2024.108241","DOIUrl":"10.1016/j.jip.2024.108241","url":null,"abstract":"<div><div>The humoral response plays a crucial role in insect defense against parasites and pathogens, typically producing antimicrobial peptides through the Toll, IMD, and Jak-STAT signaling pathways, as well as melanization via phenoloxidases. However, many studies use nonpathogenic or opportunistic organisms and often infect insects in nonnatural ways, such as piercing or injecting the pathogen into the hemocoel. The objective of this study was to examine the modulation of the main humoral pathway genes involved in the interaction between the nonmodel organism <em>Diatraea saccharalis</em> (the sugarcane borer) and different biological control agents. We identified and evaluated the expression of <em>DsDorsal</em> (Toll pathway), <em>DsRelish</em> (IMD pathway), <em>DsSTAT</em> (JAK/STAT pathway), <em>DsPPO1</em>, and <em>DsPPO2</em> (PO pathway) in larvae and pupae of <em>D. saccharalis</em> exposed or not to different biological control agents. The biocontrol agents used were: (i) the bacterium <em>Bacillus thuringiensis</em> var. <em>aizawai</em> GC-91, which is pathogenic to <em>D. saccharalis</em>; (ii) the fungus <em>Metarhizium anisopliae</em> ESALQE9 strain, which is employed to control the froghoppers of the genus <em>Mahanarva</em> in sugarcane fields, though it exhibits low virulence to <em>D. saccharalis</em>; and (iii) the generalist parasitoid <em>Tetrastichus howardi</em>. Our results demonstrate that <em>B. thuringiensis</em> at LC<sub>30</sub> induced the expression of <em>DsRelish</em> at 24 h and <em>DsSTAT</em> at 48 h after treatment initiation. In contrast, treatment with the <em>M. anisopliae</em> ESALQE9 strain reduced the levels of <em>DsDorsal</em> and <em>DsSTAT</em> 24 h post-infection compared to the control group. In larvae, <em>DsDorsal</em>, <em>DsSTAT</em>, <em>DsPPO1</em>, and <em>DsPPO2</em> were induced in response to <em>T. howardi</em>, whereas no induction was observed in pupae. Notably, no immune-related genes were modulated during the pupae-parasitoid interaction. Additionally, we provide an explanation for why <em>T. howardi</em> shows superior parasitism success in <em>D. saccharalis</em> pupae compared with larvae. The data presented here introduce novel perspectives for enhancing pest management through the utilization of biocontrol agents.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"209 ","pages":"Article 108241"},"PeriodicalIF":3.6,"publicationDate":"2024-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142791788","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 : 2024-12-03DOI: 10.1016/j.jip.2024.108239
Kayla A. Samms , Emma C. Monod , Aizah Ijaz , Sarah Au , Kristof Jenik , Tania Rodríguez-Ramos , Brian Dixon , Stephanie J. DeWitte-Orr
White spot syndrome virus (WSSV), a double stranded (ds)DNA virus, is a pathogen that causes massive mortalities in crustaceans worldwide. The present study focuses on using dsRNA to induce sequence-independent immune responses to control virus replication. DsRNA is a well characterized innate immune stimulant in vertebrates and effectively induces an antiviral state. In crustaceans, it has been shown that dsRNA containing WSSV sequences (WSSV-dsRNA) can trigger an immune response independent of RNA interference (RNAi) to mitigate disease. We hypothesized that the potency and efficacy of dsRNA-induced immunity would be enhanced using a biodegradable, cationic phytoglycogen nanoparticle, Nanodendrix (nanoparticle; NP), to deliver the dsRNA. Two in vivo studies were conducted to test the efficacy of long dsRNA as an innate immune stimulant with or without the NP in crayfish. Long dsRNA, 360–500 bp in length, was synthesized based on two WSSV sequences, viral particle 28 (VP28) and viral particle 19 (VP19) respectively. Crayfish were injected in the ventral sinus with WSSV-dsRNA (VP28 or VP19 sequence) either in complex with the NP or alone. High molecular weight (HMW) poly inosinic: polycytidylic acid (poly IC), a synthetic viral dsRNA mimic, was used as a positive control. In the negative control groups, crayfish were injected with either phosphate buffered saline or NP alone. These studies found WSSV-dsRNA could enhance hemocyte numbers, nitric oxide levels and phenoloxidase activity. This enhancement was more effective than when using HMW poly IC. Finally, the nanoparticle did not increase dsRNA’s immune activation capability, but it did reduce dsRNA’s toxicity. Further studies may help determine the efficacy of these treatments as immune stimulants for preventing pathogenic outbreaks in the invertebrate aquaculture industry.
{"title":"Sequence independent immune effects of white spot syndrome virus (WSSV) dsRNA complexed with phytoglycogen nanoparticles in freshwater crayfish","authors":"Kayla A. Samms , Emma C. Monod , Aizah Ijaz , Sarah Au , Kristof Jenik , Tania Rodríguez-Ramos , Brian Dixon , Stephanie J. DeWitte-Orr","doi":"10.1016/j.jip.2024.108239","DOIUrl":"10.1016/j.jip.2024.108239","url":null,"abstract":"<div><div>White spot syndrome virus (WSSV), a double stranded (ds)DNA virus, is a pathogen that causes massive mortalities in crustaceans worldwide. The present study focuses on using dsRNA to induce sequence-independent immune responses to control virus replication. DsRNA is a well characterized innate immune stimulant in vertebrates and effectively induces an antiviral state. In crustaceans, it has been shown that dsRNA containing WSSV sequences (WSSV-dsRNA) can trigger an immune response independent of RNA interference (RNAi) to mitigate disease. We hypothesized that the potency and efficacy of dsRNA-induced immunity would be enhanced using a biodegradable, cationic phytoglycogen nanoparticle, Nanodendrix (nanoparticle; NP), to deliver the dsRNA. Two <em>in vivo</em> studies were conducted to test the efficacy of long dsRNA as an innate immune stimulant with or without the NP in crayfish. Long dsRNA, 360–500 bp in length, was synthesized based on two WSSV sequences, viral particle 28 (VP28) and viral particle 19 (VP19) respectively. Crayfish were injected in the ventral sinus with WSSV-dsRNA (VP28 or VP19 sequence) either in complex with the NP or alone. High molecular weight (HMW) poly inosinic: polycytidylic acid (poly IC), a synthetic viral dsRNA mimic, was used as a positive control. In the negative control groups, crayfish were injected with either phosphate buffered saline or NP alone. These studies found WSSV-dsRNA could enhance hemocyte numbers, nitric oxide levels and phenoloxidase activity. This enhancement was more effective than when using HMW poly IC. Finally, the nanoparticle did not increase dsRNA’s immune activation capability, but it did reduce dsRNA’s toxicity. Further studies may help determine the efficacy of these treatments as immune stimulants for preventing pathogenic outbreaks in the invertebrate aquaculture industry.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"209 ","pages":"Article 108239"},"PeriodicalIF":3.6,"publicationDate":"2024-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142785947","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-12-02DOI: 10.1016/j.jip.2024.108242
Natasha Sant Anna Iwanicki , Isabella Alice Gotti , Italo Delalibera Jr. , Henrik H. De Fine Licht
Generalist pathogens with a broad host range encounter many different host environments. Such generalist pathogens are often highly versatile and adjust their expressed phenotype to the host being infected. Species in the fungal genus Metarhizium (Hypocreales: Clavicipitaceae) occupy various ecological niches, including plant rhizosphere symbionts, soil saprophytes, and insect pathogens with applications in biological control of pests. The species M. anisopliae is highly diverse combining the capability of association with plant roots and infection of a broad range of arachnid and insect hosts, from agricultural pests to vectors of human disease. It is among the most studied and applied biological control agents worldwide. Here, we investigate the phenotypic plasticity and differential gene expression of M. anisopliae blastospores during infection of different insect hosts. First, the virulence of M. anisopliae blastospores was evaluated against Tenebrio molitor (Coleoptera: Tenebrionidae), Spodoptera frugiperda (Lepidoptera: Noctuidae), Gryllus assimilis (Orthoptera: Gryllidae), and Apis mellifera (Hymenoptera: Apidae). Second, the percentage of appressorium formation on the membranous wings of the four hosts was determined, and third, the fungal transcriptome profile during penetration on the hosts was analyzed. Our findings reveal that M. anisopliae blastospores exhibit high virulence against Tenebrio molitor, with significantly higher appressorium formation on beetle wings compared to the other three tested insects. We also document distinct gene expression patterns in M. anisopliae blastospores during insect infection of T. molitor, S. frugiperda, and A. mellifera, with notable variations observed in G. assimilis. These differences are associated with the expression of enzymes involved in the degradation of specific compounds present in each insect wing, as well as hydrophobins, destruxins, and specialized metabolites related to virulence. The study emphasizes the differences in fungal gene expression during infection of the four insect orders and highlights the virulence-related genes specific to each infective process.
{"title":"Host-specific patterns of virulence and gene expression profiles of the broad-host-range entomopathogenic fungus Metarhizium anisopliae","authors":"Natasha Sant Anna Iwanicki , Isabella Alice Gotti , Italo Delalibera Jr. , Henrik H. De Fine Licht","doi":"10.1016/j.jip.2024.108242","DOIUrl":"10.1016/j.jip.2024.108242","url":null,"abstract":"<div><div>Generalist pathogens with a broad host range encounter many different host environments. Such generalist pathogens are often highly versatile and adjust their expressed phenotype to the host being infected. Species in the fungal genus <em>Metarhizium</em> (Hypocreales: Clavicipitaceae) occupy various ecological niches, including plant rhizosphere symbionts, soil saprophytes, and insect pathogens with applications in biological control of pests. The species <em>M. anisopliae</em> is highly diverse combining the capability of association with plant roots and infection of a broad range of arachnid and insect hosts, from agricultural pests to vectors of human disease. It is among the most studied and applied biological control agents worldwide. Here, we investigate the phenotypic plasticity and differential gene expression of <em>M. anisopliae</em> blastospores during infection of different insect hosts. First, the virulence of <em>M. anisopliae</em> blastospores was evaluated against <em>Tenebrio molitor</em> (Coleoptera: Tenebrionidae)<em>, Spodoptera frugiperda</em> (Lepidoptera: Noctuidae), <em>Gryllus assimilis</em> (Orthoptera: Gryllidae), and <em>Apis mellifera</em> (Hymenoptera: Apidae)<em>.</em> Second, the percentage of appressorium formation on the membranous wings of the four hosts was determined, and third, the fungal transcriptome profile during penetration on the hosts was analyzed. Our findings reveal that <em>M. anisopliae</em> blastospores exhibit high virulence against <em>Tenebrio molitor</em>, with significantly higher appressorium formation on beetle wings compared to the other three tested insects. We also document distinct gene expression patterns in <em>M. anisopliae</em> blastospores during insect infection of <em>T. molitor, S. frugiperda,</em> and <em>A. mellifera</em>, with notable variations observed in <em>G. assimilis</em>. These differences are associated with the expression of enzymes involved in the degradation of specific compounds present in each insect wing, as well as hydrophobins, destruxins, and specialized metabolites related to virulence. The study emphasizes the differences in fungal gene expression during infection of the four insect orders and highlights the virulence-related genes specific to each infective process.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"209 ","pages":"Article 108242"},"PeriodicalIF":3.6,"publicationDate":"2024-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142780358","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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