Biological control using nematophagous fungi such as Duddingtonia flagrans offers a sustainable strategy to reduce gastrointestinal nematode populations in grazing animals. However, large-scale application requires efficient chlamydospore production without compromising fungal viability and efficacy. This study aimed to optimize a solid culture medium enriched with meso-inositol and mannitol to enhance chlamydospore production and assess its impact on nematode predatory capacity for their subsequent use as biological control agent. Different formulations were tested by supplementing Glucose Sabouraud Agar with various concentrations of meso-inositol (1.1–2 %) and mannitol (2–5 %). Cultures were incubated for up to 35 days, and chlamydospores were extracted and quantified under optical microscope at 3, 4, and 5 weeks of culturing. The highest yield – 6.95 × 10⁷ chlamydospores/plate - was obtained using 2 % meso-inositol and 5 % mannitol after 35 days of incubation at 27 ± 0.5 °C and 70 ± 5 % RH. In vitro fungal predatory activity against gastrointestinal nematode larvae from naturally parasitized sheep was maintained across all formulation treatments, with parasite larval reduction exceeding 70 % (p < 0.0001), indicating that the optimized medium did not impair nematophagous efficacy. These findings contribute to the development of scale-up fungal culture strategies for producing commercially-based D. flagrans suitable for its incorporation into integrated control programs of livestock parasites.
{"title":"Development of an optimized culture medium using meso-inositol and mannitol to maximize chlamydospore production of Duddingtonia flagrans","authors":"Manuela Southwell , Milagros Junco , Silvina Fernández , Gisele Bernat , Sara Zegbi , Inés Guerrero , Federica Sagües","doi":"10.1016/j.vetpar.2025.110613","DOIUrl":"10.1016/j.vetpar.2025.110613","url":null,"abstract":"<div><div>Biological control using nematophagous fungi such as <em>Duddingtonia flagrans</em> offers a sustainable strategy to reduce gastrointestinal nematode populations in grazing animals. However, large-scale application requires efficient chlamydospore production without compromising fungal viability and efficacy. This study aimed to optimize a solid culture medium enriched with meso-inositol and mannitol to enhance chlamydospore production and assess its impact on nematode predatory capacity for their subsequent use as biological control agent. Different formulations were tested by supplementing Glucose Sabouraud Agar with various concentrations of meso-inositol (1.1–2 %) and mannitol (2–5 %). Cultures were incubated for up to 35 days, and chlamydospores were extracted and quantified under optical microscope at 3, 4, and 5 weeks of culturing. The highest yield – 6.95 × 10⁷ chlamydospores/plate - was obtained using 2 % meso-inositol and 5 % mannitol after 35 days of incubation at 27 ± 0.5 °C and 70 ± 5 % RH. <em>In vitro</em> fungal predatory activity against gastrointestinal nematode larvae from naturally parasitized sheep was maintained across all formulation treatments, with parasite larval reduction exceeding 70 % (p < 0.0001), indicating that the optimized medium did not impair nematophagous efficacy. These findings contribute to the development of scale-up fungal culture strategies for producing commercially-based <em>D. flagrans</em> suitable for its incorporation into integrated control programs of livestock parasites.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"340 ","pages":"Article 110613"},"PeriodicalIF":2.2,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145096577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-18DOI: 10.1016/j.vetpar.2025.110611
Gisela Neira , Roberto Mera y Sierra , Dayana Godoy , Lorena Logarzo , Mariana Gonzalez , Silvana Scarcella
Fascioliasis is a globally distributed parasitic disease caused by Fasciola hepatica and Fasciola gigantica, transmitted by freshwater snails of the family Lymnaeidae. Although environmental factors influence the distribution of these intermediate hosts, the role of water physicochemical parameters remains underexplored. This study evaluated the relationship between water quality and lymnaeid snail presence in endemic areas of western Argentina. A total of 53 aquatic sites were surveyed, with lymnaeid snails detected in 33 (62.26 %). Water samples were categorized as positive or negative based on snail presence and F. hepatica detection in livestock from the associated farm. Significant differences were observed in electrical conductivity, total dissolved solids, pH, total hardness, calcium, sodium, and sulfate concentrations. Correlation analysis revealed a consistent negative association between lymnaeid presence and most parameters, particularly calcium and total hardness. Notably, snails were found in sites with water temperatures as low as 2.8°C, challenging previous assumptions about thermal limits for survival. Consistently with these environmental findings, coprological analysis of 1936 animals showed a strong association between snail presence and F. hepatica infection: all farms in snail-positive areas tested positive (19/19), while none in snail-negative areas did (0/10) (Fisher’s exact test, p < 0.001). Overall prevalence in snail-positive farms was 36.48 %, with species-specific rates ranging from 25.51 % in cattle to 50.91 % in goats. These findings provide actionable insights for environmental surveillance and fascioliasis control strategies in endemic regions, especially under changing climatic conditions.
{"title":"Influence of water quality parameters on lymnaeid snail distribution and their relationship with fasciolosis transmission","authors":"Gisela Neira , Roberto Mera y Sierra , Dayana Godoy , Lorena Logarzo , Mariana Gonzalez , Silvana Scarcella","doi":"10.1016/j.vetpar.2025.110611","DOIUrl":"10.1016/j.vetpar.2025.110611","url":null,"abstract":"<div><div>Fascioliasis is a globally distributed parasitic disease caused by <em>Fasciola hepatica</em> and <em>Fasciola gigantica,</em> transmitted by freshwater snails of the family Lymnaeidae<em>.</em> Although environmental factors influence the distribution of these intermediate hosts, the role of water physicochemical parameters remains underexplored. This study evaluated the relationship between water quality and lymnaeid snail presence in endemic areas of western Argentina. A total of 53 aquatic sites were surveyed, with lymnaeid snails detected in 33 (62.26 %). Water samples were categorized as positive or negative based on snail presence and <em>F. hepatica</em> detection in livestock from the associated farm. Significant differences were observed in electrical conductivity, total dissolved solids, pH, total hardness, calcium, sodium, and sulfate concentrations. Correlation analysis revealed a consistent negative association between lymnaeid presence and most parameters, particularly calcium and total hardness. Notably, snails were found in sites with water temperatures as low as 2.8°C, challenging previous assumptions about thermal limits for survival. Consistently with these environmental findings, coprological analysis of 1936 animals showed a strong association between snail presence and <em>F. hepatica</em> infection: all farms in snail-positive areas tested positive (19/19), while none in snail-negative areas did (0/10) (Fisher’s exact test, p < 0.001). Overall prevalence in snail-positive farms was 36.48 %, with species-specific rates ranging from 25.51 % in cattle to 50.91 % in goats. These findings provide actionable insights for environmental surveillance and fascioliasis control strategies in endemic regions, especially under changing climatic conditions.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"340 ","pages":"Article 110611"},"PeriodicalIF":2.2,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145096576","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-16DOI: 10.1016/j.vetpar.2025.110610
Beatrice Mercaldo, Maria Chiara Alterisio, Antonio Bosco, Antonio Di Loria, Elena Ciccone, Sergio Esposito, Laura Rinaldi, Paolo Ciaramella, Jacopo Guccione
This study aimed to evaluate the effectiveness of ultrasound (US) in sheep compared with the necropsy (gold standard) as an in-vivo monitoring tool used in a long-term control program of cystic echinococcosis (CE). The study involved 10-randomly-selected farms, divided into a Treated-Group (TG, n = 5, receiving a control protocol for CE) and a Control-Group (CG, n = 5, no protocol), enrolled over 6-years in an endemic area of southern Italy. All sheep of ten farms destined for slaughter underwent liver and lung US followed by necropsy, during the study period. From a total of 1’175 sheep, 50.0 % (593/1’175) belonged to the TG and 49.5 % (582/1’175) to the CG were enrolled. Overall, the US showed a Sensitivity (SE) of 87.9 %, a Specificity (SP) of 72.3 % as well as a positive- (PPV) and negative-predictive-value (NPV), and Accuracy of 74.4 %, 86.7 %, and 79.7 %, respectively. A moderate Cohen’s Kappa-Coefficient (K=0.599) were also detected between the two techniques. The generalised linear mixed model showed that distribution (p < 0.001) and type of lesion (p < 0.001) significantly influence the US performance. The higher SP and PPV in CG likely result from a greater disease prevalence and presence of older lesions; the higher NPV in TG might reflect the beneficial effects of the control program and lower disease exposure. Although technical and logistical challenges have to be addressed for its use, a US conscious integration into continuous surveillance program might promote the early in vivo identification of infected farms, limiting necropsy dependence for CE monitoring.
本研究旨在评价超声（US）与尸检（金标准）作为囊性包虫病（CE）长期控制项目体内监测工具的有效性。该研究涉及10个随机选择的农场，分为治疗组（TG, n = 5，接受CE的对照方案）和对照组（CG, n = 5，无方案），在意大利南部的一个流行区注册6年。在研究期间，10个屠宰场的所有羊都进行了肝脏和肺部检查，然后进行了尸检。在1'175只羊中，50.0 %（593/1'175）属于TG， 49.5 %（582/1'175）属于CG。总体而言，美国的敏感性（SE）为87.9% %，特异性（SP）为72.3 %，阳性预测值（PPV）和阴性预测值（NPV），准确性分别为74.4 %，86.7 %和79.7% %。两种方法之间的Cohen’s kappa系数（K=0.599）均为中等。广义线性混合模型表明，分布(p
{"title":"Effectiveness of ultrasound in sheep as a monitoring tool for the long-term control of cystic echinococcosis","authors":"Beatrice Mercaldo, Maria Chiara Alterisio, Antonio Bosco, Antonio Di Loria, Elena Ciccone, Sergio Esposito, Laura Rinaldi, Paolo Ciaramella, Jacopo Guccione","doi":"10.1016/j.vetpar.2025.110610","DOIUrl":"10.1016/j.vetpar.2025.110610","url":null,"abstract":"<div><div>This study aimed to evaluate the effectiveness of ultrasound (US) in sheep compared with the necropsy (gold standard) as an <em>in-vivo</em> monitoring tool used in a long-term control program of cystic echinococcosis (CE). The study involved 10-randomly-selected farms, divided into a Treated-Group (TG, n = 5, receiving a control protocol for CE) and a Control-Group (CG, n = 5, no protocol), enrolled over 6-years in an endemic area of southern Italy. All sheep of ten farms destined for slaughter underwent liver and lung US followed by necropsy, during the study period. From a total of 1’175 sheep, 50.0 % (593/1’175) belonged to the TG and 49.5 % (582/1’175) to the CG were enrolled. Overall, the US showed a Sensitivity (SE) of 87.9 %, a Specificity (SP) of 72.3 % as well as a positive- (PPV) and negative-predictive-value (NPV), and Accuracy of 74.4 %, 86.7 %, and 79.7 %, respectively. A moderate Cohen’s Kappa-Coefficient (<em>K=</em>0.599) were also detected between the two techniques. The generalised linear mixed model showed that distribution (p < 0.001) and type of lesion (p < 0.001) significantly influence the US performance. The higher SP and PPV in CG likely result from a greater disease prevalence and presence of older lesions; the higher NPV in TG might reflect the beneficial effects of the control program and lower disease exposure. Although technical and logistical challenges have to be addressed for its use, a US conscious integration into continuous surveillance program might promote the early <em>in vivo</em> identification of infected farms, limiting necropsy dependence for CE monitoring.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"340 ","pages":"Article 110610"},"PeriodicalIF":2.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145091600","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-16DOI: 10.1016/j.vetpar.2025.110609
Pedro Henrique Cotrin Rodrigues , Nelson Henrique de Almeida Curi , Elizabete Pereira Barbosa , Guilherme Ramos Demétrio , Ana Maria Oliveira Paschoal , Rodrigo Lima Massara , Adriano Garcia Chiarello , Marcelo Passamani , Camila Stefanie Fonseca de Oliveira , Daniel Sobreira Rodrigues , Júlia Angélica Gonçalves Silveira
Sentinel hosts are species that are readily observable and more likely than others to be exposed to vector and vector borne pathogens. Surveys of domestic dogs are often utilized to assess tick distribution and tick-borne diseases. Free-ranging habits and poor health management probably increase infectious diseases in rural dogs. The Atlantic Forest biome occurs in some South American countries and is a major center of endemism, highly threatened by deforestation. This study describes the prevalence for ectoparasites and tick-borne pathogens in 323 dogs living at 144 households around six Atlantic Forest conservation units. There were found ticks of the genus Rhipicephalus (11.5 %) and Amblyomma (23.5 %), fleas of the genus Ctenocephalides (44.3 %), Xenopsylla (1.24 %), and Tunga (0.3 %), and lice Trichodectes canis (0,9 %). Blood smear analysis revealed a prevalence of 34.6 % of at least one of the following parasite taxa: Hepatozoon (22.3 %), Babesia vogeli (4.9 %), Ehrlichia canis (3.1 %), and Anaplasma platys (2.8 %). In seven dogs (2.2 %), coinfections were detected microscopically. Serologic tests revealed seroreaction in 47 % of dogs, and the presence at considerable seroprevalence of E. canis (34.3 %), Anaplasma phagocytophilum (23.8 %) and B. vogeli (10.5 %). Serological coinfections were detected in 19.5 % of dogs. The molecular analyses performed using samples from 157 ectoparasites that parasitized the dogs, showed a prevalence of 1.27 % positive for order Piroplasmida, 1.27 % for Ehrlichia spp., 3.82 % for Anaplasma spp., and 22.93 % for Rickettsia spp. The findings highlight the importance of rural dogs as sentinels of diseases for wildlife and humans in Brazil, and the need for straightforward control actions such as treatment against ectoparasites with sufficient coverage, and responsible domestic animal ownership enforcement.
{"title":"Rural dogs (Canis familiaris) as sentinels to ectoparasites and tick borne pathogens around six Atlantic Forest conservation units in Brazil","authors":"Pedro Henrique Cotrin Rodrigues , Nelson Henrique de Almeida Curi , Elizabete Pereira Barbosa , Guilherme Ramos Demétrio , Ana Maria Oliveira Paschoal , Rodrigo Lima Massara , Adriano Garcia Chiarello , Marcelo Passamani , Camila Stefanie Fonseca de Oliveira , Daniel Sobreira Rodrigues , Júlia Angélica Gonçalves Silveira","doi":"10.1016/j.vetpar.2025.110609","DOIUrl":"10.1016/j.vetpar.2025.110609","url":null,"abstract":"<div><div>Sentinel hosts are species that are readily observable and more likely than others to be exposed to vector and vector borne pathogens. Surveys of domestic dogs are often utilized to assess tick distribution and tick-borne diseases. Free-ranging habits and poor health management probably increase infectious diseases in rural dogs. The Atlantic Forest biome occurs in some South American countries and is a major center of endemism, highly threatened by deforestation. This study describes the prevalence for ectoparasites and tick-borne pathogens in 323 dogs living at 144 households around six Atlantic Forest conservation units. There were found ticks of the genus <em>Rhipicephalus</em> (11.5 %) and <em>Amblyomma</em> (23.5 %), fleas of the genus <em>Ctenocephalides</em> (44.3 %), <em>Xenopsylla</em> (1.24 %), and <em>Tunga</em> (0.3 %), and lice <em>Trichodectes canis</em> (0,9 %). Blood smear analysis revealed a prevalence of 34.6 % of at least one of the following parasite taxa: <em>Hepatozoon</em> (22.3 %), <em>Babesia vogeli</em> (4.9 %), <em>Ehrlichia canis</em> (3.1 %), and <em>Anaplasma platys</em> (2.8 %). In seven dogs (2.2 %), coinfections were detected microscopically. Serologic tests revealed seroreaction in 47 % of dogs, and the presence at considerable seroprevalence of <em>E. canis</em> (34.3 %), <em>Anaplasma phagocytophilum</em> (23.8 %) and <em>B. vogeli</em> (10.5 %). Serological coinfections were detected in 19.5 % of dogs. The molecular analyses performed using samples from 157 ectoparasites that parasitized the dogs, showed a prevalence of 1.27 % positive for order Piroplasmida, 1.27 % for <em>Ehrlichia</em> spp., 3.82 % for <em>Anaplasma</em> spp., and 22.93 % for <em>Rickettsia</em> spp. The findings highlight the importance of rural dogs as sentinels of diseases for wildlife and humans in Brazil, and the need for straightforward control actions such as treatment against ectoparasites with sufficient coverage, and responsible domestic animal ownership enforcement.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"340 ","pages":"Article 110609"},"PeriodicalIF":2.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145087486","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-13DOI: 10.1016/j.vetpar.2025.110608
Alexander Dindial , Sean Monaghan , Jay Haywood , Kevin McLean , Dorota Androscuk , Kim Thompson , William Roy , James Bron
The salmon louse (Lepeophtheirus salmonis (Krøyer, 1837)) is a caligid ectoparasite of salmonids that feeds on host blood, mucus, and skin. While secreted virulence factors from later life stages have been studied, the protein composition of secretory and excretory products (SEPs) from copepodids, the initial infectious stage of L. salmonis, remains uncharacterized.
Copepodids were hatched and incubated at 10°C until 7 days post-hatch. Batches (n = 4) were then exposed to either 0.45 μm filtered seawater or 0.1 mg mL⁻¹ isophorone to stimulate SEP production. Adult males and females (n = 2 replicates) were similarly treated for comparison. SEPs were filtered, precipitated, trypsin-digested, and analyzed via LC-MS/MS. Proteins were identified using an L. salmonis database and further analyzed with SignalP and InterPro.
In total, 433 distinct proteins were detected in copepodid samples (mean 95.5 ± 146.74), and 117 in adult samples (mean 56 ± 12.70). Signal peptide analysis revealed 164 copepodid and 69 adult proteins as secretory. Among adults, 31 secretory proteins were female-specific and 10 male-specific. Twenty-one secretory proteins were shared across life stages, including 8 proteases, 2 protease inhibitors, and 2 uncharacterized proteins. Of proteins with GO annotations, 75 % were involved in proteolysis and 50 % localized extracellularly. However, secretory profiles differed markedly between life stages. Notably, 67 % of adult-specific secretory proteins were extracellular versus 30.7 % in copepodids. Copepodid and adult SEPs also contained 23 and 4 unique uncharacterized proteins, respectively.
These findings highlight a complex repertoire of copepodid SEPs potentially involved in host invasion and immunomodulation, providing new targets for therapeutic development.
{"title":"Investigation of proteins identified in the secretory and excretory products (SEPs) of the infectious copepodid stage of the salmon louse Lepeophtheirus salmonis","authors":"Alexander Dindial , Sean Monaghan , Jay Haywood , Kevin McLean , Dorota Androscuk , Kim Thompson , William Roy , James Bron","doi":"10.1016/j.vetpar.2025.110608","DOIUrl":"10.1016/j.vetpar.2025.110608","url":null,"abstract":"<div><div>The salmon louse (<em>Lepeophtheirus salmonis</em> (Krøyer, 1837)) is a caligid ectoparasite of salmonids that feeds on host blood, mucus, and skin. While secreted virulence factors from later life stages have been studied, the protein composition of secretory and excretory products (SEPs) from copepodids, the initial infectious stage of <em>L. salmonis</em>, remains uncharacterized.</div><div>Copepodids were hatched and incubated at 10°C until 7 days post-hatch. Batches (n = 4) were then exposed to either 0.45 μm filtered seawater or 0.1 mg mL⁻¹ isophorone to stimulate SEP production. Adult males and females (n = 2 replicates) were similarly treated for comparison. SEPs were filtered, precipitated, trypsin-digested, and analyzed via LC-MS/MS. Proteins were identified using an <em>L. salmonis</em> database and further analyzed with SignalP and InterPro.</div><div>In total, 433 distinct proteins were detected in copepodid samples (mean 95.5 ± 146.74), and 117 in adult samples (mean 56 ± 12.70). Signal peptide analysis revealed 164 copepodid and 69 adult proteins as secretory. Among adults, 31 secretory proteins were female-specific and 10 male-specific. Twenty-one secretory proteins were shared across life stages, including 8 proteases, 2 protease inhibitors, and 2 uncharacterized proteins. Of proteins with GO annotations, 75 % were involved in proteolysis and 50 % localized extracellularly. However, secretory profiles differed markedly between life stages. Notably, 67 % of adult-specific secretory proteins were extracellular versus 30.7 % in copepodids. Copepodid and adult SEPs also contained 23 and 4 unique uncharacterized proteins, respectively.</div><div>These findings highlight a complex repertoire of copepodid SEPs potentially involved in host invasion and immunomodulation, providing new targets for therapeutic development.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"340 ","pages":"Article 110608"},"PeriodicalIF":2.2,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145091932","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-11DOI: 10.1016/j.vetpar.2025.110607
Zhonghuai Zhang , Zhuo Lan , Yuan Gao , Hongyu Qiu , Junfeng Gao , Chunren Wang
Strongylus vulgaris, Strongylus equinus, Strongylus edentatus and Strongylus asini are important equine nematodes, classified under the genus Strongylus (subfamily Strongylinae). However, this taxonomy is controversial. Thus, the mitochondrial (mt) genome of S. edentatus collected from horses in Heilongjiang Province, China was sequenced and compared with those of closely related species to clarify the classification and phylogenetic relationships of this genus. The circular mt genome of S. edentatus was 14,765 bp long, comprising 12 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and three noncoding regions. All protein-coding genes were transcribed in the same direction and had a type 3 gene arrangement. The genome contained 22 intergenic spacers (1–146 bp) and four overlapping regions (1–14 bp). The A+T content was 78.15 %, reflecting a strong nucleotide and codon usage bias. The mt genome of S. edentatus showed higher similarity with that of S. equinus (96.3 %) than with that of S. vulgaris (76.9 %). Phylogenetic analyses showed that S. edentatus and S. equinus were classified on the same branch, forming a sister branch with S. vulgaris within Strongylinae, with S. edentatus and S. equinus having a closer phylogenetic relationship. Although traditionally assigned to Strongylinae, Strongylus and Triodontophorus did not cluster together. This study provides the first complete mt genome of S. edentatus and confirmed that S. equinus, S. vulgaris, and S. edentatus belong to the genus Strongylus, which is monophyletic, with the subfamilies Strongylinae and Cyathostominae being paraphyletic. Thus, overall, this study provides valuable data for addressing taxonomic ambiguities in the Strongylidae.
{"title":"Mitochondrial genome of Strongylus edentatus confirms monophyly of the genus Strongylus within Strongylinae","authors":"Zhonghuai Zhang , Zhuo Lan , Yuan Gao , Hongyu Qiu , Junfeng Gao , Chunren Wang","doi":"10.1016/j.vetpar.2025.110607","DOIUrl":"10.1016/j.vetpar.2025.110607","url":null,"abstract":"<div><div><em>Strongylus vulgaris</em>, <em>Strongylus equinus</em>, <em>Strongylus edentatus</em> and <em>Strongylus asini</em> are important equine nematodes, classified under the genus <em>Strongylus</em> (subfamily Strongylinae). However, this taxonomy is controversial. Thus, the mitochondrial (mt) genome of <em>S. edentatus</em> collected from horses in Heilongjiang Province, China was sequenced and compared with those of closely related species to clarify the classification and phylogenetic relationships of this genus. The circular mt genome of <em>S. edentatus</em> was 14,765 bp long, comprising 12 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and three noncoding regions. All protein-coding genes were transcribed in the same direction and had a type 3 gene arrangement. The genome contained 22 intergenic spacers (1–146 bp) and four overlapping regions (1–14 bp). The A+T content was 78.15 %, reflecting a strong nucleotide and codon usage bias. The mt genome of <em>S. edentatus</em> showed higher similarity with that of <em>S. equinus</em> (96.3 %) than with that of <em>S. vulgaris</em> (76.9 %). Phylogenetic analyses showed that <em>S. edentatus</em> and <em>S. equinus</em> were classified on the same branch, forming a sister branch with <em>S. vulgaris</em> within Strongylinae, with <em>S. edentatus</em> and <em>S. equinus</em> having a closer phylogenetic relationship. Although traditionally assigned to Strongylinae, <em>Strongylus</em> and <em>Triodontophorus</em> did not cluster together. This study provides the first complete mt genome of <em>S. edentatus</em> and confirmed that <em>S. equinus</em>, <em>S. vulgaris,</em> and <em>S. edentatus</em> belong to the genus <em>Strongylus</em>, which is monophyletic, with the subfamilies Strongylinae and Cyathostominae being paraphyletic. Thus, overall, this study provides valuable data for addressing taxonomic ambiguities in the Strongylidae.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"340 ","pages":"Article 110607"},"PeriodicalIF":2.2,"publicationDate":"2025-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145046352","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-09DOI: 10.1016/j.vetpar.2025.110605
Xiaoping Tan , Tianqiang Liu , Gaoxue Wang
Parasitic infestations remain a major challenge in aquaculture, causing substantial economic losses. Current treatments for gyrodactylosis predominantly rely on unapproved chemicals, which often exhibit suboptimal efficacy. In this study, a molecular docking approach was employed to elucidate interactions between FDA-approved drugs and key residues within the active site of Gyrodactylus kobayashii proteins. Using AutoDock Vina, we screened a library of FDA-approved compounds and identified amlodipine and colistin sulfate salt as the most potent candidates against G. kobayashii. The minimum concentrations required to achieve complete parasite eradication within 24 h were 13.0 mg/L for amlodipine and 16.0 mg/L for colistin sulfate salt. Given its lower toxicity to goldfish, amlodipine was selected for further investigation. In vitro assays demonstrated that amlodipine exhibited significant anthelmintic activity, with 18 mg/L eradicating 60.67 % of G. kobayashii within 2 h. Subsequent in vivo trials involving goldfish infected with G. kobayashii revealed that a 24-hour bath treatment with amlodipine reduced the total worm burden, achieving an EC50 value of 10.537 mg/L, and 100 % efficacy at 13.0 mg/L. Acute toxicity assays determined an LC50 of 19.653 mg/L for goldfish following 96 h of exposure to amlodipine. Furthermore, expression analyses of xenobiotic-sensitive genes (cyp1a and hsp70) indicated significant modulation post-treatment, with a gradual return to baseline levels over time. To further elucidate the mechanistic effects of amlodipine, we investigated the expression of key genes within the MAPK signaling pathway, including JNK, P53, SMAD4, JNK3, and C-JUN. Results showed that the expression of these genes was significantly influenced by both the concentration and duration of amlodipine exposure. These findings underscore the potential of virtual screening methodologies for identifying effective anthelmintic agents targeting specific proteins. This approach offers promising implications for the aquaculture industry, which has historically lagged behind other sectors in drug research and development.
{"title":"Virtual screening of FDA-approved drugs identifies amlodipine as a novel anti-Gyrodactylus kobayashii agent with therapeutic potential in aquaculture","authors":"Xiaoping Tan , Tianqiang Liu , Gaoxue Wang","doi":"10.1016/j.vetpar.2025.110605","DOIUrl":"10.1016/j.vetpar.2025.110605","url":null,"abstract":"<div><div>Parasitic infestations remain a major challenge in aquaculture, causing substantial economic losses. Current treatments for gyrodactylosis predominantly rely on unapproved chemicals, which often exhibit suboptimal efficacy. In this study, a molecular docking approach was employed to elucidate interactions between FDA-approved drugs and key residues within the active site of <em>Gyrodactylus kobayashii</em> proteins. Using AutoDock Vina, we screened a library of FDA-approved compounds and identified amlodipine and colistin sulfate salt as the most potent candidates against <em>G. kobayashii</em>. The minimum concentrations required to achieve complete parasite eradication within 24 h were 13.0 mg/L for amlodipine and 16.0 mg/L for colistin sulfate salt. Given its lower toxicity to goldfish, amlodipine was selected for further investigation. In vitro assays demonstrated that amlodipine exhibited significant anthelmintic activity, with 18 mg/L eradicating 60.67 % of <em>G. kobayashii</em> within 2 h. Subsequent in vivo trials involving goldfish infected with <em>G. kobayashii</em> revealed that a 24-hour bath treatment with amlodipine reduced the total worm burden, achieving an EC<sub>50</sub> value of 10.537 mg/L, and 100 % efficacy at 13.0 mg/L. Acute toxicity assays determined an LC<sub>50</sub> of 19.653 mg/L for goldfish following 96 h of exposure to amlodipine. Furthermore, expression analyses of xenobiotic-sensitive genes (<em>cyp1a</em> and <em>hsp70</em>) indicated significant modulation post-treatment, with a gradual return to baseline levels over time. To further elucidate the mechanistic effects of amlodipine, we investigated the expression of key genes within the MAPK signaling pathway, including <em>JNK</em>, <em>P53</em>, <em>SMAD4</em>, <em>JNK3</em>, and <em>C-JUN</em>. Results showed that the expression of these genes was significantly influenced by both the concentration and duration of amlodipine exposure. These findings underscore the potential of virtual screening methodologies for identifying effective anthelmintic agents targeting specific proteins. This approach offers promising implications for the aquaculture industry, which has historically lagged behind other sectors in drug research and development.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"340 ","pages":"Article 110605"},"PeriodicalIF":2.2,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145046343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-09DOI: 10.1016/j.vetpar.2025.110606
Muhammad Zahid Sarfaraz , Sidra Abbas , Muhammad Arfan Zaman , Muhammad Zulqarnain , Kasim Sakran Abass , Asia Parveen , Safina Kousar
Eimeria zuernii is one of the most pathogenic protozoan parasites causing coccidiosis in livestock. This study evaluated the anticoccidial potential of aqueous and methanolic extracts of herbal mixture of Syzygium cumini and Trachyspermum ammi against E. zuernii. Phytochemical analysis, in vitro sporulation inhibition (SPI), and in vivo fecal oocyst count reduction (FOCR) assays were conducted. Mass spectrometry analysis identified a higher number of phytochemicals in the methanolic extract (28) compared to the aqueous extract (16), including astragalin, malvidin 3-laminaribioside, and myricetin. In the SPI assay, the methanolic extract exhibited superior efficacy, achieving 81.33 % inhibition at 500 µg mL⁻¹ compared to 74 % for the aqueous extract and 87 % for sulphadimidine. In the FOCR trial, 130 naturally infected calves were divided into five groups (n = 26) and treated orally with methanolic extract (500 µg kg⁻¹, 2 mg kg⁻¹, and 4 mg kg⁻¹), sulphadimidine (1.2 mL kg⁻¹), or water. The 4 mg kg⁻¹ methanolic extract dose achieved the highest efficacy, with a 91.3 % reduction in oocyst counts on day 15 post-treatment. Molecular docking of phytochemicals against lactate dehydrogenase (LDH) revealed astragalin as the top binder, showing strong affinity comparable to the natural inhibitor NAD. These findings validate the traditional use of these plant extracts as natural remedies for E. zuernii in calves, particularly in resource-limited settings, and highlight astragalin as a potential lead compound for future drug development. Large-scale field trials are recommended to assess the commercial applicability of these extracts.
{"title":"Phytochemical profiling and anticoccidial activity of Syzygium cumini and Trachyspermum ammi extracts against Eimeria zuernii: Integrated in vitro, in vivo, and in silico approaches","authors":"Muhammad Zahid Sarfaraz , Sidra Abbas , Muhammad Arfan Zaman , Muhammad Zulqarnain , Kasim Sakran Abass , Asia Parveen , Safina Kousar","doi":"10.1016/j.vetpar.2025.110606","DOIUrl":"10.1016/j.vetpar.2025.110606","url":null,"abstract":"<div><div><em>Eimeria zuernii</em> is one of the most pathogenic protozoan parasites causing coccidiosis in livestock. This study evaluated the anticoccidial potential of aqueous and methanolic extracts of herbal mixture of <em>Syzygium cumini</em> and <em>Trachyspermum ammi</em> against <em>E. zuernii</em>. Phytochemical analysis, <em>in vitro</em> sporulation inhibition (SPI), and <em>in vivo</em> fecal oocyst count reduction (FOCR) assays were conducted. Mass spectrometry analysis identified a higher number of phytochemicals in the methanolic extract (28) compared to the aqueous extract (16), including astragalin, malvidin 3-laminaribioside, and myricetin. In the SPI assay, the methanolic extract exhibited superior efficacy, achieving 81.33 % inhibition at 500 µg mL⁻¹ compared to 74 % for the aqueous extract and 87 % for sulphadimidine. In the FOCR trial, 130 naturally infected calves were divided into five groups (n = 26) and treated orally with methanolic extract (500 µg kg⁻¹, 2 mg kg⁻¹, and 4 mg kg⁻¹), sulphadimidine (1.2 mL kg⁻¹), or water. The 4 mg kg⁻¹ methanolic extract dose achieved the highest efficacy, with a 91.3 % reduction in oocyst counts on day 15 post-treatment. Molecular docking of phytochemicals against lactate dehydrogenase (LDH) revealed astragalin as the top binder, showing strong affinity comparable to the natural inhibitor NAD. These findings validate the traditional use of these plant extracts as natural remedies for <em>E. zuernii</em> in calves, particularly in resource-limited settings, and highlight astragalin as a potential lead compound for future drug development. Large-scale field trials are recommended to assess the commercial applicability of these extracts.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"340 ","pages":"Article 110606"},"PeriodicalIF":2.2,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145046351","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The excessive use of acaricides in the management of the tick Rhipicephalus (Boophilus) microplus has favored the development of resistance in the parasites, in addition to causing environmental contamination. In this context, biological control using entomopathogenic nematodes appears as a promising alternative. This study evaluated the efficacy of two isolates, Heterorhabditis bacteriophora (HB EN01) and Steinernema rarum (PAM 25), in the in vitro control of R. (B.) microplus. Two experiments were conducted with different concentrations of nematodes. In the first, fifteen engorged females were exposed to suspensions containing 100, 200 and 300 infective juveniles (IJs) per female. The second experiment was carried out in soil substrate, with a concentration of 200 IJs per female. Three application systems were tested: S1 (females placed on the substrate before application), S2 (suspension applied before the introduction of females) and S3 (females placed in a voile bag on the substrate). For S. rarum (PAM 25), there was no significant difference in oviposition inhibition, but larval hatching was lower at concentrations of 200 and 300 IJs/female compared to 100 (p < 0.05). H. bacteriophora (HB EN01) showed a significant reduction in oviposition only between concentrations of 100 and 300 IJs/female (p < 0.05), with no variation in larval hatching or efficacy. On the soil substrate, both isolates showed efficacy above 80 %, except for S. rarum in the voile system (60 %). Treated females showed darkening and dehydration from the 4th day onwards. The control group remained healthy. The results indicate that EPNs are an effective alternative in the control of R. (B.) microplus.
{"title":"Effect of Heterorhabditis bacteriophora (HB EN01) and Steinernema rarum (PAM 25) nematodes on the control of cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)","authors":"E.R. Souza, J.G.C. Orozco, F.C. Duarte, L.C. Torres, M.C. Mendes","doi":"10.1016/j.vetpar.2025.110604","DOIUrl":"10.1016/j.vetpar.2025.110604","url":null,"abstract":"<div><div>The excessive use of acaricides in the management of the tick <em>Rhipicephalus</em> (<em>Boophilus</em>) <em>microplus</em> has favored the development of resistance in the parasites, in addition to causing environmental contamination. In this context, biological control using entomopathogenic nematodes appears as a promising alternative. This study evaluated the efficacy of two isolates, <em>Heterorhabditis bacteriophora</em> (HB EN01) and <em>Steinernema rarum</em> (PAM 25), in the in vitro control of <em>R</em>. (<em>B</em>.) <em>microplus</em>. Two experiments were conducted with different concentrations of nematodes. In the first, fifteen engorged females were exposed to suspensions containing 100, 200 and 300 infective juveniles (IJs) per female. The second experiment was carried out in soil substrate, with a concentration of 200 IJs per female. Three application systems were tested: S1 (females placed on the substrate before application), S2 (suspension applied before the introduction of females) and S3 (females placed in a voile bag on the substrate). For <em>S</em>. <em>rarum</em> (PAM 25), there was no significant difference in oviposition inhibition, but larval hatching was lower at concentrations of 200 and 300 IJs/female compared to 100 (p < 0.05). <em>H</em>. <em>bacteriophora</em> (HB EN01) showed a significant reduction in oviposition only between concentrations of 100 and 300 IJs/female (p < 0.05), with no variation in larval hatching or efficacy. On the soil substrate, both isolates showed efficacy above 80 %, except for <em>S. rarum</em> in the voile system (60 %). Treated females showed darkening and dehydration from the 4th day onwards. The control group remained healthy. The results indicate that EPNs are an effective alternative in the control of <em>R</em>. (<em>B.) microplus</em>.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"340 ","pages":"Article 110604"},"PeriodicalIF":2.2,"publicationDate":"2025-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145091597","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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