The immune response of insects to pathogenic micro-organisms infection usually exhibits significant sex differences. Understanding this sexual dimorphism elucidates how immunity interplays with reproductive strategy, metabolic status and environmental factors. In this study, we showed female Drosophila melanogaster (fruit fly) survived better than males when infected with Metarhizium anisopliae CQMa421. This sex-specific survival difference depends on the Toll pathway, which is critical for innate immunity. We systematically compared innate immunity, including humoral immunity, cellular immunity, and melanization, between infected male and female flies. Female flies displayed a faster humoral immune response, characterized by earlier upregulation of antimicrobial peptides (AMPs) expression in both fat body and gut. On the other hand, male flies exhibited a stronger early cellular immune response, marked by increased expression of genes associated with hemocyte clotting and phagocytosis. While male flies upregulated melanization related Prophenoloxidase (PPO) earlier than female flies, both sexes significantly increased phenoloxidase (PO) activity after fungal infection. Fungal load quantification revealed that CQMa421 proliferated more quickly in females than in males. Collectively, our data indicate that each sex exhibits advantages in specific immune response, and that sex-biased survival outcome arises from the integration of multiple host defense mechanisms.
{"title":"Sexual dimorphism in the immune response of Drosophila melanogaster to the entomopathogenic fungus Metarhizium anisopliae.","authors":"Yulian Peng, Jinliang Lyu, Qinwei Li, Petros Ligoxygakis, Yuxian Xia, Qi Xiao","doi":"10.1016/j.jip.2025.108422","DOIUrl":"10.1016/j.jip.2025.108422","url":null,"abstract":"<p><p>The immune response of insects to pathogenic micro-organisms infection usually exhibits significant sex differences. Understanding this sexual dimorphism elucidates how immunity interplays with reproductive strategy, metabolic status and environmental factors. In this study, we showed female Drosophila melanogaster (fruit fly) survived better than males when infected with Metarhizium anisopliae CQMa421. This sex-specific survival difference depends on the Toll pathway, which is critical for innate immunity. We systematically compared innate immunity, including humoral immunity, cellular immunity, and melanization, between infected male and female flies. Female flies displayed a faster humoral immune response, characterized by earlier upregulation of antimicrobial peptides (AMPs) expression in both fat body and gut. On the other hand, male flies exhibited a stronger early cellular immune response, marked by increased expression of genes associated with hemocyte clotting and phagocytosis. While male flies upregulated melanization related Prophenoloxidase (PPO) earlier than female flies, both sexes significantly increased phenoloxidase (PO) activity after fungal infection. Fungal load quantification revealed that CQMa421 proliferated more quickly in females than in males. Collectively, our data indicate that each sex exhibits advantages in specific immune response, and that sex-biased survival outcome arises from the integration of multiple host defense mechanisms.</p>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":" ","pages":"108422"},"PeriodicalIF":2.4,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144804261","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 : 2025-10-30DOI: 10.1016/j.jip.2025.108480
Daniella-Mari White , Emmanouela Karaveti , Vasileios Bakopoulos
Fish farming may pose a risk to adjacent octopus farms due to pathogen transmission. Moreover, the immune defense mechanisms of cephalopods are still not fully understood. This study aimed to determine changes in total protein concentration and hemolysis activity of Octopus vulgaris hemolymph, after intramuscular (IM) or intravenous (IV) challenges with aquaculture fish pathogens (either Photobacterium damselae subsp. piscicida or damselae or Vibrio alginolyticus or anguillarum O1) at two temperatures (21 ± 0.5 ℃ and 24 ± 0.5 ℃).
Results showed that Octopus vulgaris exhibited a mean total protein concentration of 173.93 ± 69.37 mg/mL across all experimental conditions, markedly exceeding values reported for other mollusks, such as the bivalves Chamelea gallina (0.75–1.66 mg/mL) and Mytilus galloprovincialis (0.59–1.60 mg/mL). Patterns of total protein concentration, related to the genera of the pathogen used for the challenges, were observed. Four-way ANOVA revealed significant main effects of bacterium (F(3, 144) = 54.360, p < 0.001) and temperature (F(1, 144) = 10.014, p = 0.002) on total protein, along with multiple significant interaction effects, including bacterium × temperature, route × time, and bacterium × route × temperature × time (all p < 0.001).
Hemolysis remained at low levels across both experimental temperatures, challenge routes, and pathogens, not exceeding 25 % in any case. Values above 15 % and up to 20 % were recorded in specific conditions, such as Photobacterium damselae subsp. damselae at 24 ± 0.5 °C on Day 3 in CIM-, IM-, and IV- control and challenged groups respectivelly; V. alginolyticus at 24 ± 0.5 °C on Day 3 in IM-challenged groups; and Vibrio anguillarum O1 at 21 ± 0.5 °C on Day 3 and Day 7 in IM-challenged groups. ANOVA for hemolytic activity showed significant main effects of bacterium (F(3, 144) = 22.032, p < 0.001) and temperature (F(1, 144) = 4.083, p = 0.045), with multiple significant interactions, including bacterium × temperature, route × time, and bacterium × route × temperature × time (all p < 0.001). These results indicate that the route of challenge may play a major role in hemolysis activity, with temperature and time post-challenge also exerting significant effects, possibly through a complex synergistic interaction.
Our results may assist in elucidating common octopus defense mechanisms against common fish pathogens and provide important information to the scientific community and the marine aquaculture sector.
{"title":"Assessing the impact of gram-negative bacteria on the common octopus, in relation to rising sea temperature: a study of total protein concentration and hemolysis activity in hemolymph","authors":"Daniella-Mari White , Emmanouela Karaveti , Vasileios Bakopoulos","doi":"10.1016/j.jip.2025.108480","DOIUrl":"10.1016/j.jip.2025.108480","url":null,"abstract":"<div><div>Fish farming may pose a risk to adjacent octopus farms due to pathogen transmission. Moreover, the immune defense mechanisms of cephalopods are still not fully understood. This study aimed to determine changes in total protein concentration and hemolysis activity of <em>Octopus vulgaris</em> hemolymph, after intramuscular (IM) or intravenous (IV) challenges with aquaculture fish pathogens (either <em>Photobacterium damselae</em> subsp. <em>piscicida</em> or <em>damselae</em> or <em>Vibrio alginolyticus</em> or <em>anguillarum</em> O1) at two temperatures (21 ± 0.5 ℃ and 24 ± 0.5 ℃).</div><div>Results showed that <em>Octopus vulgaris</em> exhibited a mean total protein concentration of 173.93 ± 69.37 mg/mL across all experimental conditions, markedly exceeding values reported for other mollusks, such as the bivalves <em>Chamelea gallina</em> (0.75–1.66 mg/mL) and <em>Mytilus galloprovincialis</em> (0.59–1.60 mg/mL). Patterns of total protein concentration, related to the genera of the pathogen used for the challenges, were observed. Four-way ANOVA revealed significant main effects of bacterium (F(3, 144) = 54.360, p < 0.001) and temperature (F(1, 144) = 10.014, p = 0.002) on total protein, along with multiple significant interaction effects, including bacterium × temperature, route × time, and bacterium × route × temperature × time (all p < 0.001).</div><div>Hemolysis remained at low levels across both experimental temperatures, challenge routes, and pathogens, not exceeding 25 % in any case. Values above 15 % and up to 20 % were recorded in specific conditions, such as <em>Photobacterium damselae</em> subsp. <em>damselae</em> at 24 ± 0.5 °C on Day 3 in CIM-, IM-, and IV- control and challenged groups respectivelly; <em>V. alginolyticus</em> at 24 ± 0.5 °C on Day 3 in IM-challenged groups; and <em>Vibrio anguillarum</em> O1 at 21 ± 0.5 °C on Day 3 and Day 7 in IM-challenged groups. ANOVA for hemolytic activity showed significant main effects of bacterium (F(3, 144) = 22.032, p < 0.001) and temperature (F(1, 144) = 4.083, p = 0.045), with multiple significant interactions, including bacterium × temperature, route × time, and bacterium × route × temperature × time (all p < 0.001). These results indicate that the route of challenge may play a major role in hemolysis activity, with temperature and time post-challenge also exerting significant effects, possibly through a complex synergistic interaction.</div><div>Our results may assist in elucidating common octopus defense mechanisms against common fish pathogens and provide important information to the scientific community and the marine aquaculture sector.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108480"},"PeriodicalIF":2.4,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145421971","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 : 2025-10-30DOI: 10.1016/j.jip.2025.108481
Junjiang Liu , Lu Zhang , Xin Jia , Jinyu Yang , Mengqiang Wang
Acute hepatopancreatic necrosis disease (AHPND) poses a major threat to global shrimp aquaculture, especially impacting Penaeus vannamei. Given the high cost of prevention and the absence of effective treatments, the most efficient way to control AHPND is through early detection to quickly identify the pathogen. This study presents the development and optimization of a low-cost, portable diagnostic platform utilizing loop-mediated isothermal amplification (LAMP) for the rapid detection of AHPND. This platform integrates fluorescence detection with a smartphone-compatible device, providing a convenient and effective solution for on-site diagnosis. We also screened fluorescent nucleic acid dyes with optimal adaptability and developed an efficient, user-friendly tool to improve the performance of the isothermal amplification method. In addition, a deep learning-based infection detection algorithm was developed for automated diagnosis. The visualization technology showed high specificity and sensitivity for detecting AHPND in shrimp samples, with a detection limit of 1 copies/µL. These results highlight the potential of this method in resource-limited environments and provide a valuable tool for early detection and management of AHPND in shrimp aquaculture.
{"title":"Construction and optimization of a LAMP-based diagnostic platform for acute hepatopancreatic necrosis disease in Penaeus vannamei","authors":"Junjiang Liu , Lu Zhang , Xin Jia , Jinyu Yang , Mengqiang Wang","doi":"10.1016/j.jip.2025.108481","DOIUrl":"10.1016/j.jip.2025.108481","url":null,"abstract":"<div><div>Acute hepatopancreatic necrosis disease (AHPND) poses a major threat to global shrimp aquaculture, especially impacting <em>Penaeus vannamei</em>. Given the high cost of prevention and the absence of effective treatments, the most efficient way to control AHPND is through early detection to quickly identify the pathogen. This study presents the development and optimization of a low-cost, portable diagnostic platform utilizing loop-mediated isothermal amplification (LAMP) for the rapid detection of AHPND. This platform integrates fluorescence detection with a smartphone-compatible device, providing a convenient and effective solution for on-site diagnosis. We also screened fluorescent nucleic acid dyes with optimal adaptability and developed an efficient, user-friendly tool to improve the performance of the isothermal amplification method. In addition, a deep learning-based infection detection algorithm was developed for automated diagnosis. The visualization technology showed high specificity and sensitivity for detecting AHPND in shrimp samples, with a detection limit of 1 copies/µL. These results highlight the potential of this method in resource-limited environments and provide a valuable tool for early detection and management of AHPND in shrimp aquaculture.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108481"},"PeriodicalIF":2.4,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145416886","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 : 2025-10-30DOI: 10.1016/j.jip.2025.108486
Lin-Rui Chang , Cai-Yi Chen , Fan Yang , Chuan-Yu He , Fan Li , Ting Li , Ling-Jun Si , Dong-Chun Yan
Ecytonucleospora (=Enterocytozoon) hepatopenaei (EHP) is an intracellular parasitic microsporidian, Infectious hypodermal and haematopoietic necrosis virus (IHHNV) is a linear single-stranded DNA virus. Both of them are widespread pathogens in shrimp farming, causing slow growth in shrimp and leading to significant economic losses. Litopenaeus vannamei (L. vannamei), the dominant species in shrimp farming, can be co-infected by EHP and IHHNV. According to our prior research, EHP and IHHNV exhibit synergistic interactions during infection. However, the underlying mechanisms of their co-infection remain unclear. In this study, L. vannamei from four experimental groups (EHP infected group, IHHNV infected group, EHP and IHHNV co-infected group, healthy group) were compared by transcriptome sequencing to analyze the synergistic pathogenic mechanisms of IHHNV and EHP. The results showed that co-infection induced more pronounced changes in the expression of genes related to metabolism, growth, and immunity compared to single infections. Specifically, EHP infection had a greater impact on shrimp physiology and immune responses, suggesting a dominant role in co-infection. Meanwhile, IHHNV infection disrupted metabolic processes, inhibited molting, and exacerbated the immune suppression caused by EHP. The synergistic pathogenic mechanism of IHHNV and EHP may involve EHP weakening the immune defenses of L. vannamei, facilitating IHHNV infection and proliferation. Additionally, the metabolic dysregulation induced by IHHNV may further enhance consumption of cellular nutrients and energy by EHP. These findings preliminarily reveal the synergistic pathogenic mechanism of EHP and IHHNV in L. vannamei, offering a theoretical foundation for understanding disease outbreaks in shrimp farming and guiding strategies against multi-pathogen infections.
胞核孢子虫（=Enterocytozoon）肝原孢子虫（EHP）是一种细胞内寄生微孢子虫，感染皮下和造血坏死病毒（IHHNV）是线性单链DNA病毒。这两种病毒都是虾类养殖中广泛存在的病原体，导致虾类生长缓慢，造成重大经济损失。凡纳滨对虾（Litopenaeus vannamei， L. vannamei）是对虾养殖中的优势种，可同时感染EHP和IHHNV。根据我们之前的研究，EHP和IHHNV在感染过程中表现出协同作用。然而，其共同感染的潜在机制尚不清楚。本研究通过转录组测序对4个实验组（EHP感染组、IHHNV感染组、EHP与IHHNV共感染组、健康组）的凡纳梅L.进行比较，分析IHHNV与EHP的协同致病机制。结果表明，与单一感染相比，联合感染诱导了与代谢、生长和免疫相关基因的表达发生了更明显的变化。具体而言，EHP感染对虾的生理和免疫反应有更大的影响，表明在共感染中起主导作用。同时，IHHNV感染破坏了代谢过程，抑制了蜕皮，加剧了EHP引起的免疫抑制。IHHNV与EHP的协同致病机制可能与EHP削弱南美嗜血杆菌的免疫防御，促进IHHNV感染和增殖有关。此外，IHHNV诱导的代谢失调可能进一步增加EHP对细胞营养和能量的消耗。研究结果初步揭示了南美对虾EHP与IHHNV的协同致病机制，为了解对虾养殖中的疾病暴发和指导多病原体感染策略提供了理论依据。
{"title":"Synergistic pathogenic mechanisms of EHP and IHHNV co-infection in Litopenaeus vannamei: Insights from RNA-seq analysis","authors":"Lin-Rui Chang , Cai-Yi Chen , Fan Yang , Chuan-Yu He , Fan Li , Ting Li , Ling-Jun Si , Dong-Chun Yan","doi":"10.1016/j.jip.2025.108486","DOIUrl":"10.1016/j.jip.2025.108486","url":null,"abstract":"<div><div><em>Ecytonucleospora</em> (=<em>Enterocytozoon</em>) <em>hepatopenaei</em> (EHP) is an intracellular parasitic microsporidian, Infectious hypodermal and haematopoietic necrosis virus (IHHNV) is a linear single-stranded DNA virus. Both of them are widespread pathogens in shrimp farming, causing slow growth in shrimp and leading to significant economic losses. <em>Litopenaeus vannamei</em> (<em>L. vannamei</em>), the dominant species in shrimp farming, can be co-infected by EHP and IHHNV. According to our prior research, EHP and IHHNV exhibit synergistic interactions during infection. However, the underlying mechanisms of their co-infection remain unclear. In this study, <em>L. vannamei</em> from four experimental groups (EHP infected group, IHHNV infected group, EHP and IHHNV co-infected group, healthy group) were compared by transcriptome sequencing to analyze the synergistic pathogenic mechanisms of IHHNV and EHP. The results showed that co-infection induced more pronounced changes in the expression of genes related to metabolism, growth, and immunity compared to single infections. Specifically, EHP infection had a greater impact on shrimp physiology and immune responses, suggesting a dominant role in co-infection. Meanwhile, IHHNV infection disrupted metabolic processes, inhibited molting, and exacerbated the immune suppression caused by EHP. The synergistic pathogenic mechanism of IHHNV and EHP may involve EHP weakening the immune defenses of <em>L. vannamei</em>, facilitating IHHNV infection and proliferation. Additionally, the metabolic dysregulation induced by IHHNV may further enhance consumption of cellular nutrients and energy by EHP. These findings preliminarily reveal the synergistic pathogenic mechanism of EHP and IHHNV in <em>L. vannamei</em>, offering a theoretical foundation for understanding disease outbreaks in shrimp farming and guiding strategies against multi-pathogen infections.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108486"},"PeriodicalIF":2.4,"publicationDate":"2025-10-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145426465","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 : 2025-10-29DOI: 10.1016/j.jip.2025.108485
Xiaolin Cai , Chunxiu Pang , Fuchang Zhuo , Bo Hu , Xuehua Huang , Jiaxing Huang , Yuming Lu
Melissococcus plutonius, the agent of European foulbrood (EFB), has been well studied in Apis mellifera but its epidemiology in Apis cerana remains less understood. We surveyed 37 apiaries across Guangxi, China, and detected M. plutonius in 32.4%, 10.8%, and 27.0% of larvae, adult bees, and honey samples, respectively, all originating from asymptomatic colonies. Duplex PCR revealed frequent co-detection of typical and atypical strains. Viable isolates were recovered from 58.3% of PCR-positive larval samples. Multi-locus sequence typing (MLST) showed six isolates (T1, X16, L20, Q21, Q22, L32) clustering with the atypical reference strain DAT561, and one isolate (P6) clustering with the typical reference strain ATCC 35311. Our findings highlight the high prevalence and strain diversity of M. plutonius in A. cerana in subtropical Guangxi and emphasize the need for region-specific surveillance strategies.
{"title":"High prevalence and strain diversity of Melissococcus plutonius in Apis cerana in Guangxi, China","authors":"Xiaolin Cai , Chunxiu Pang , Fuchang Zhuo , Bo Hu , Xuehua Huang , Jiaxing Huang , Yuming Lu","doi":"10.1016/j.jip.2025.108485","DOIUrl":"10.1016/j.jip.2025.108485","url":null,"abstract":"<div><div><em>Melissococcus plutonius</em>, the agent of European foulbrood (EFB), has been well studied in <em>Apis mellifera</em> but its epidemiology in <em>Apis cerana</em> remains less understood. We surveyed 37 apiaries across Guangxi, China, and detected <em>M. plutonius</em> in 32.4%, 10.8%, and 27.0% of larvae, adult bees, and honey samples, respectively, all originating from asymptomatic colonies. Duplex PCR revealed frequent co-detection of typical and atypical strains. Viable isolates were recovered from 58.3% of PCR-positive larval samples. Multi-locus sequence typing (MLST) showed six isolates (T1, X16, L20, Q21, Q22, L32) clustering with the atypical reference strain DAT561, and one isolate (<em>P</em>6) clustering with the typical reference strain ATCC 35311. Our findings highlight the high prevalence and strain diversity of <em>M. plutonius</em> in <em>A. cerana</em> in subtropical Guangxi and emphasize the need for region-specific surveillance strategies.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108485"},"PeriodicalIF":2.4,"publicationDate":"2025-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145421952","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 : 2025-10-29DOI: 10.1016/j.jip.2025.108484
Vincent Prayugo , Alexandria N. Payne , Bryony C. Bonning , Adam G. Dolezal
Honey bees are often exposed to and infected by multiple viruses, including deformed wing virus (DWV) and Israeli acute paralysis virus (IAPV). These pathogens are major contributors to colony failure, threatening the supply of pollination services in agriculture. However, understanding and modeling the infection and transmission dynamics of these viruses is hampered by gaps in knowledge about their infection timelines, specifically how virus load changes post-exposure. Moreover, while honey bees frequently experience co-occurring infections from multiple viruses, little is known about how co-infection affects virus-induced mortality and replication dynamics. We hypothesized that, while both DWV and IAPV are known to be highly infectious and can cause honey bee mortality, each presents a distinct infection timeline and peak infection intensity time window. In addition, we hypothesized that co-infection would result in increased mortality compared to individual virus infections due to increased pathogen-induced stress. To test these hypotheses, we exposed day-old honey bee workers to DWV, IAPV, or a combination of DWV and IAPV via carefully controlled experimental injections and tracked mortality and virus levels over time. After exposure, bees were maintained in the laboratory for 10 days and sampled daily for virus quantification. We observed a similar response pattern to both honey bee viruses in the first 24 h post-infection, where virus levels rapidly increased following inoculation. While DWV infection had persistently high virus levels and a delayed mortality response after peak DWV load was achieved, IAPV infection was rapidly followed by either mortality or a decline in virus load as bees recovered from infection. Co-infected bees showed some variation in mortality, although there were no significant differences in virus load between DWV or IAPV within co-infected bees compared to bees infected with the individual viruses. These results help answer fundamental questions related to the pathology of DWV and IAPV within honey bees that help clarify how these pathogens interact and persist within honey bee colonies.
{"title":"Infection timelines and co-infection effects of Israeli acute paralysis virus and deformed wing virus in the honey bee (Apis mellifera)","authors":"Vincent Prayugo , Alexandria N. Payne , Bryony C. Bonning , Adam G. Dolezal","doi":"10.1016/j.jip.2025.108484","DOIUrl":"10.1016/j.jip.2025.108484","url":null,"abstract":"<div><div>Honey bees are often exposed to and infected by multiple viruses, including deformed wing virus (DWV) and Israeli acute paralysis virus (IAPV). These pathogens are major contributors to colony failure, threatening the supply of pollination services in agriculture. However, understanding and modeling the infection and transmission dynamics of these viruses is hampered by gaps in knowledge about their infection timelines, specifically how virus load changes post-exposure. Moreover, while honey bees frequently experience co-occurring infections from multiple viruses, little is known about how co-infection affects virus-induced mortality and replication dynamics. We hypothesized that, while both DWV and IAPV are known to be highly infectious and can cause honey bee mortality, each presents a distinct infection timeline and peak infection intensity time window. In addition, we hypothesized that co-infection would result in increased mortality compared to individual virus infections due to increased pathogen-induced stress. To test these hypotheses, we exposed day-old honey bee workers to DWV, IAPV, or a combination of DWV and IAPV via carefully controlled experimental injections and tracked mortality and virus levels over time. After exposure, bees were maintained in the laboratory for 10 days and sampled daily for virus quantification. We observed a similar response pattern to both honey bee viruses in the first 24 h post-infection, where virus levels rapidly increased following inoculation. While DWV infection had persistently high virus levels and a delayed mortality response after peak DWV load was achieved, IAPV infection was rapidly followed by either mortality or a decline in virus load as bees recovered from infection. Co-infected bees showed some variation in mortality, although there were no significant differences in virus load between DWV or IAPV within co-infected bees compared to bees infected with the individual viruses. These results help answer fundamental questions related to the pathology of DWV and IAPV within honey bees that help clarify how these pathogens interact and persist within honey bee colonies.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108484"},"PeriodicalIF":2.4,"publicationDate":"2025-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145416885","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 : 2025-10-28DOI: 10.1016/j.jip.2025.108487
Maria Cristina Bruno , Sonia Endrizzi , Andrea Basso , Valentina Paolini , Tobia Pretto
One of the causes of the decline in distribution and abundance of the endangered white-clawed crayfish Austropotamobius pallipes complex throughout Europe is the invasion of alien crayfish, and the associated spread of infectious diseases, primarily the crayfish plague caused by Aphanomyces astaci. Another relevant disease is microsporidiosis (porcelain disease), caused by Astathelohania contejeani and Nosema austropotamobii. Between 2021 and 2024, we conducted a monitoring survey, aimed at mapping the distribution of A. astaci, A. contejeani and N. austropotamobii in wild populations of A. pallipes and in two non-indigenous species in Trentino (North-East Italy). We applied a non-invasive sampling method (cuticular swabs) in 33 populations of A. pallipes, 2 populations of Procambarus clarkii and 4 populations of Faxonius limosus, to investigate the presence of A. astaci. Aphanomyces astaci was detected in eight thriving populations of A. pallipes, and the presence of a low virulence genotype (genotype group A) was confirmed in one of them. Aphanomyces astaci was detected in one of the two populations of P. clarkii, as well as in one of the four populations of F. limosus. No mortality outbreaks in populations of A. pallipes were recorded in Trentino during the study period. Specimens with macroscopic signs of porcelain disease were found in thirteen populations of A. pallipes, abdominal muscle tissues were collected and subjected to molecular evaluation; microsporidiosis was detected in all the tested crayfish: seven populations were infected only by A. contejeani, and in six populations both microsporidia were detected. Crayfish from one population of P. clarkii and one of F. limosus were tested for microsporidiosis as well, but the results were negative. Based on the presence of chronically infected but thriving populations across the Trentino territory, we provide suggestions for the management of A. pallipes populations in conservation actions which require the movement of specimens (translocations, rearing in captivity).
{"title":"Crayfish plague and microsporidiosis occurrence in wild populations of the white-clawed crayfish Austropotamobius pallipes complex in Trentino (North-East Italy)","authors":"Maria Cristina Bruno , Sonia Endrizzi , Andrea Basso , Valentina Paolini , Tobia Pretto","doi":"10.1016/j.jip.2025.108487","DOIUrl":"10.1016/j.jip.2025.108487","url":null,"abstract":"<div><div>One of the causes of the decline in distribution and abundance of the endangered white-clawed crayfish <em>Austropotamobius pallipes</em> complex throughout Europe is the invasion of alien crayfish, and the associated spread of infectious diseases, primarily the crayfish plague caused by <em>Aphanomyces astaci</em>. Another relevant disease is microsporidiosis (porcelain disease), caused by <em>Astathelohania contejeani</em> and <em>Nosema austropotamobii</em>. Between 2021 and 2024, we conducted a monitoring survey, aimed at mapping the distribution of <em>A. astaci</em>, <em>A. contejeani</em> and <em>N. austropotamobii</em> in wild populations of <em>A. pallipes</em> and in two non-indigenous species in Trentino (North-East Italy). We applied a non-invasive sampling method (cuticular swabs) in 33 populations of <em>A. pallipes</em>, 2 populations of <em>Procambarus clarkii</em> and 4 populations of <em>Faxonius limosus</em>, to investigate the presence of <em>A. astaci</em>. <em>Aphanomyces astaci</em> was detected in eight thriving populations of <em>A. pallipes</em>, and the presence of a low virulence genotype (genotype group A) was confirmed in one of them. <em>Aphanomyces astaci</em> was detected in one of the two populations of <em>P. clarkii</em>, as well as in one of the four populations of <em>F. limosus</em>. No mortality outbreaks in populations of <em>A. pallipes</em> were recorded in Trentino during the study period. Specimens with macroscopic signs of porcelain disease were found in thirteen populations of <em>A. pallipes,</em> abdominal muscle tissues were collected and subjected to molecular evaluation; microsporidiosis was detected in all the tested crayfish: seven populations were infected only by <em>A. contejeani</em>, and in six populations both microsporidia were detected. Crayfish from one population of <em>P. clarkii</em> and one of <em>F. limosus</em> were tested for microsporidiosis as well, but the results were negative. Based on the presence of chronically infected but thriving populations across the Trentino territory, we provide suggestions for the management of <em>A. pallipes</em> populations in conservation actions which require the movement of specimens (translocations, rearing in captivity).</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108487"},"PeriodicalIF":2.4,"publicationDate":"2025-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145409162","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 : 2025-10-28DOI: 10.1016/j.jip.2025.108488
Ge Jiang , Xianping Fan , Jie Cheng , Yi Qiao , Xiaotong Wang , Yijun Liu , Xiaohui Cao , Shen Hui
Acute hepatopancreatic necrosis disease (AHPND), caused by the toxin‐producing Vibrio sp., has become a serious threat to shrimp aquaculture. Egg yolk immunoglobulin(IgY) has the advantage of low cost and good protection in the treatment of diseases caused by specific pathogens in crustaceans. This study evaluated the protective effect of IgY against AHPND infection in Penaeus vannamei. IgY was isolated from eggs laid by hens immunized with recombinant PirAB toxin, which prepared with the PirAB gene as the target gene. Freeze-dried egg powders with anti‐PirAB‐IgY were mixed with basal diets at 20 % and 10 % concentrations and IgY from non‐immunized hen (control‐IgY) was mixed with basal diets at 20 % concentrations and used to prefeed shrimp 3 days before the bacterial challenge test. Survival rates of the challenged shrimp fed the anti‐PirAB‐IgY(20 %), anti‐PirAB‐IgY(10 %), and control‐IgY diets were 70 %, 50 % and 13 %, respectively. The parameters including β-1,3-d-glucan-binding protein and Heat shock proteins 70 (Hsp70) had increased in the experimental groups fed with anti‐PirAB‐IgY compared with the positive control group, and the expression levels of crustin were significantly(P < 0.05) higher in the positive control group than in the other groups. This study provides an effective prophylactic method against AHPND infection in shrimp.
{"title":"Oral administration of egg yolk immunoglobulin (IgY) in Penaeus vannamei to combat against VpAHPND infections","authors":"Ge Jiang , Xianping Fan , Jie Cheng , Yi Qiao , Xiaotong Wang , Yijun Liu , Xiaohui Cao , Shen Hui","doi":"10.1016/j.jip.2025.108488","DOIUrl":"10.1016/j.jip.2025.108488","url":null,"abstract":"<div><div>Acute hepatopancreatic necrosis disease (AHPND), caused by the toxin‐producing <em>Vibrio</em> sp., has become a serious threat to shrimp aquaculture. Egg yolk immunoglobulin(IgY) has the advantage of low cost and good protection in the treatment of diseases caused by specific pathogens in crustaceans. This study evaluated the protective effect of IgY against AHPND infection in <em>Penaeus vannamei</em>. IgY was isolated from eggs laid by hens immunized with recombinant PirAB toxin, which prepared with the <em>PirAB</em> gene as the target gene. Freeze-dried egg powders with anti‐PirAB‐IgY were mixed with basal diets at 20 % and 10 % concentrations and IgY from non‐immunized hen (control‐IgY) was mixed with basal diets at 20 % concentrations and used to prefeed shrimp 3 days before the bacterial challenge test. Survival rates of the challenged shrimp fed the anti‐PirAB‐IgY(20 %), anti‐PirAB‐IgY(10 %), and control‐IgY diets were 70 %, 50 % and 13 %, respectively. The parameters including β-1,3-<span>d</span>-glucan-binding protein and Heat shock proteins 70 (Hsp70) had increased in the experimental groups fed with anti‐PirAB‐IgY compared with the positive control group, and the expression levels of crustin were significantly(<em>P</em> < 0.05) higher in the positive control group than in the other groups. This study provides an effective prophylactic method against AHPND infection in shrimp.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108488"},"PeriodicalIF":2.4,"publicationDate":"2025-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145409139","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 : 2025-10-27DOI: 10.1016/j.jip.2025.108479
Li-Guo Yang , Lei Feng , Li-Yan Ma , Xue-Feng Song , Jia-Yuan Xu , Bian-Bian Zhang , Wen-Hong Fang , Xin-Cang Li
MicroRNAs (miRNAs) are known to modulate various immune responses and metabolic processes by targeting and silencing specific genes in invertebrates. However, the specific roles of miRNAs in the context of microsporidial infection in the shrimp Penaeus vannamei have not been well-defined. This study aimed to elucidate the functions of miRNAs in shrimp during the invasion by the microsporidian pathogen Ecytonucleospora hepatopenaei (EHP). To achieve this, we collected shrimp that had confirmed EHP infections and conducted a comprehensive transcriptional analysis. High-throughput sequencing data yielded 15,964,804 clean reads from the hepatopancreas of healthy shrimp and 14,301,330 clean reads from EHP-infected counterparts. From these samples, a total of 59 distinct miRNAs were identified. Notably, the expression levels of 31 miRNAs, with 16 being down-regulated and 15 up-regulated, were significantly altered in the hepatopancreas of EHP-infected shrimp when compared to the controls. The differentially expressed miRNAs were subjected to Gene Ontology (GO) analysis and target gene prediction to determine their biological relevance. The findings indicated that the miRNAs with altered expression are predominantly associated with immune response pathways and metabolic alterations, including those involved in the Toll-like receptor signaling pathway (Ko04620); the Toll and Imd signaling pathways (Ko04624); protein digestion and absorption (Ko04974); and the regulation of aldosterone synthesis and secretion (Ko04925). Collectively, our study advances our understanding of miRNAs’ roles within the innate immune system of shrimp and identifies potential novel targets for the management and prevention of EHP infections.
{"title":"Dysregulation of hepatopancreatic microRNAs in Penaeus vannamei in response to Ecytonucleospora hepatopenaei (EHP) infection: Implications for innate immunity and metabolic function","authors":"Li-Guo Yang , Lei Feng , Li-Yan Ma , Xue-Feng Song , Jia-Yuan Xu , Bian-Bian Zhang , Wen-Hong Fang , Xin-Cang Li","doi":"10.1016/j.jip.2025.108479","DOIUrl":"10.1016/j.jip.2025.108479","url":null,"abstract":"<div><div>MicroRNAs (miRNAs) are known to modulate various immune responses and metabolic processes by targeting and silencing specific genes in invertebrates. However, the specific roles of miRNAs in the context of microsporidial infection in the shrimp <em>Penaeus vannamei</em> have not been well-defined. This study aimed to elucidate the functions of miRNAs in shrimp during the invasion by the microsporidian pathogen <em>Ecytonucleospora hepatopenaei</em> (EHP). To achieve this, we collected shrimp that had confirmed EHP infections and conducted a comprehensive transcriptional analysis. High-throughput sequencing data yielded 15,964,804 clean reads from the hepatopancreas of healthy shrimp and 14,301,330 clean reads from EHP-infected counterparts. From these samples, a total of 59 distinct miRNAs were identified. Notably, the expression levels of 31 miRNAs, with 16 being down-regulated and 15 up-regulated, were significantly altered in the hepatopancreas of EHP-infected shrimp when compared to the controls. The differentially expressed miRNAs were subjected to Gene Ontology (GO) analysis and target gene prediction to determine their biological relevance. The findings indicated that the miRNAs with altered expression are predominantly associated with immune response pathways and metabolic alterations, including those involved in the Toll-like receptor signaling pathway (Ko04620); the Toll and Imd signaling pathways (Ko04624); protein digestion and absorption (Ko04974); and the regulation of aldosterone synthesis and secretion (Ko04925). Collectively, our study advances our understanding of miRNAs’ roles within the innate immune system of shrimp and identifies potential novel targets for the management and prevention of EHP infections.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108479"},"PeriodicalIF":2.4,"publicationDate":"2025-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145401093","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 : 2025-10-27DOI: 10.1016/j.jip.2025.108483
Shangning Yang , Zhichu Huang , Ruike Wei , Dandan Liu , Xiaoling Su , Huoqing Zheng
The Israeli acute paralysis virus (IAPV), widespread in honeybee (Apis mellifera) colonies, causes paralysis, tremors, and eventual mortality, while strongly associated with colony collapse disorder (CCD). Currently, there are no widely approved antiviral therapies for IAPV infection; however, several natural compounds have shown promising antiviral efficacy. In this study, we evaluated the effects of eight natural compounds (myrcene, citral, menthol, chlorogenic acid, resveratrol, curcumin, quercetin, and tea polyphenols) on IAPV-infected honeybees. Bees were artificially or naturally infected with IAPV and subsequently fed sucrose solution supplemented with these compounds. Quantitative RT-PCR analysis revealed that curcumin was the most effective compound in reducing viral load. After seven days of treatment with 10 µM curcumin, IAPV levels decreased by 46.2 % in artificially infected bees and 84.0 % in naturally infected bees. Transcriptomic analysis showed that curcumin upregulated genes involved in nutritional metabolism (e.g., Vg) and the RNA interference (RNAi) pathways (e.g., dicer, hsp90), which play a critical role in antiviral defense. Interestingly, it downregulated antimicrobial peptide genes (hymenoptaecin, abaecin, defensin 1). These findings demonstrated that curcumin mediated a strategic immune reconfiguration in honeybees, concentrating more resources on the RNAi pathway, while reducing energetically costly antimicrobial peptide synthesis. This optimized antiviral defense, combined with improved vitellogenin expression, suggests a metabolic efficiency adaptation that preferentially directs resources toward more effective IAPV countermeasures. Our results suggest that curcumin is a dual-action therapeutic candidate that simultaneously strengthens crucial antiviral defenses and maintains host vitality, offering significant promise for sustainable honeybee health management.
{"title":"Antiviral activity of curcumin against Israeli acute paralysis virus in Apis mellifera: Screening and mechanistic study","authors":"Shangning Yang , Zhichu Huang , Ruike Wei , Dandan Liu , Xiaoling Su , Huoqing Zheng","doi":"10.1016/j.jip.2025.108483","DOIUrl":"10.1016/j.jip.2025.108483","url":null,"abstract":"<div><div>The Israeli acute paralysis virus (IAPV), widespread in honeybee (<em>Apis mellifera</em>) colonies, causes paralysis, tremors, and eventual mortality, while strongly associated with colony collapse disorder (CCD). Currently, there are no widely approved antiviral therapies for IAPV infection; however, several natural compounds have shown promising antiviral efficacy. In this study, we evaluated the effects of eight natural compounds (myrcene, citral, menthol, chlorogenic acid, resveratrol, curcumin, quercetin, and tea polyphenols) on IAPV-infected honeybees. Bees were artificially or naturally infected with IAPV and subsequently fed sucrose solution supplemented with these compounds. Quantitative RT-PCR analysis revealed that curcumin was the most effective compound in reducing viral load. After seven days of treatment with 10 µM curcumin, IAPV levels decreased by 46.2 % in artificially infected bees and 84.0 % in naturally infected bees. Transcriptomic analysis showed that curcumin upregulated genes involved in nutritional metabolism (e.g., <em>Vg</em>) and the RNA interference (RNAi) pathways (e.g., <em>dicer</em>, <em>hsp90</em>), which play a critical role in antiviral defense. Interestingly, it downregulated antimicrobial peptide genes (<em>hymenoptaecin</em>, <em>abaecin</em>, <em>defensin 1</em>). These findings demonstrated that curcumin mediated a strategic immune reconfiguration in honeybees, concentrating more resources on the RNAi pathway, while reducing energetically costly antimicrobial peptide synthesis. This optimized antiviral defense, combined with improved <em>vitellogenin</em> expression, suggests a metabolic efficiency adaptation that preferentially directs resources toward more effective IAPV countermeasures. Our results suggest that curcumin is a dual-action therapeutic candidate that simultaneously strengthens crucial antiviral defenses and maintains host vitality, offering significant promise for sustainable honeybee health management.</div></div>","PeriodicalId":16296,"journal":{"name":"Journal of invertebrate pathology","volume":"214 ","pages":"Article 108483"},"PeriodicalIF":2.4,"publicationDate":"2025-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145401113","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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