Pub Date : 2026-01-14DOI: 10.1016/j.fawpar.2026.e00315
Zhao Li , Tao Li , Lian-Tao Yang , Cai-Qin Deng , Qi-Xin Liu , Qin-Zhang , Ling Wu , Yue Sun , Feng-Cai Zou , Xue Zhou , Qi-Shuai Liu
Toxoplasma gondii is a significant foodborne parasite. However, the precise thermal conditions required to inactivate its tissue cysts in meat remain poorly defined. This study systematically determined the effects of temperature (45–70 °C) and time (10–30 min) on cyst viability. Cysts treated under each condition were orally administered to susceptible C57BL/6J mice, and infectivity was comprehensively assessed through survival, clinical signs, serology (IgG), qPCR, and histopathology. Results demonstrated that treatment at 60 °C for 10 min or under more stringent conditions completely abolished infectivity, as evidenced by 100% survival, the absence of specific antibodies, and the non-detection of parasite DNA or lesions in tissues. Thus, 60 °C for 10 min is established as a critical inactivation threshold, providing a definitive reference for developing science-based thermal processing guidelines to enhance meat safety.
{"title":"Bioassay of the infectivity of heat-treated Toxoplasma gondii cysts in susceptible C57BL/6J mice","authors":"Zhao Li , Tao Li , Lian-Tao Yang , Cai-Qin Deng , Qi-Xin Liu , Qin-Zhang , Ling Wu , Yue Sun , Feng-Cai Zou , Xue Zhou , Qi-Shuai Liu","doi":"10.1016/j.fawpar.2026.e00315","DOIUrl":"10.1016/j.fawpar.2026.e00315","url":null,"abstract":"<div><div><em>Toxoplasma gondii</em> is a significant foodborne parasite. However, the precise thermal conditions required to inactivate its tissue cysts in meat remain poorly defined. This study systematically determined the effects of temperature (45–70 °C) and time (10–30 min) on cyst viability. Cysts treated under each condition were orally administered to susceptible C57BL/6J mice, and infectivity was comprehensively assessed through survival, clinical signs, serology (IgG), qPCR, and histopathology. Results demonstrated that treatment at 60 °C for 10 min or under more stringent conditions completely abolished infectivity, as evidenced by 100% survival, the absence of specific antibodies, and the non-detection of parasite DNA or lesions in tissues. Thus, 60 °C for 10 min is established as a critical inactivation threshold, providing a definitive reference for developing science-based thermal processing guidelines to enhance meat safety.</div></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"42 ","pages":"Article e00315"},"PeriodicalIF":3.1,"publicationDate":"2026-01-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146038123","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-07DOI: 10.1016/j.fawpar.2026.e00316
Si Si Ru , Wen Li , Jie Hao , Cheng Yue Cao , Li Ma , Xi Zhang
Metagenomic next-generation sequencing (mNGS) technology offers substantial advantages in parasite detection; however, we still know very little about its diagnostic value for Spirometra mansoni infection. In this study, mNGS technology was used to analyse faecal samples and blood samples from cats infected with S. mansoni, as well as tissue samples and blood samples from mice infected with the plerocercoid larvae of S. mansoni. Moreover, polymerase chain reaction (PCR) was employed to validate the mNGS results. The diagnostic value of mNGS for S. mansoni infection was systematically evaluated. The mNGS results revealed that the read counts of S. mansoni in the cat faeces (CF) samples were 301,497 (CF1), 1,330,549 (CF2), 1,181,162 (CF3), and 0 (CF0), with relative abundances of 3.17%, 16.64%, 13.14%, and 0%, respectively. In the mouse tissue (MT) samples, the read counts of S. mansoni were 10,791 (MT1), 438 (MT2), 3697 (MT3), and 10 (MT0), with relative abundances of 67.21%, 3.65%, 21.12%, and 0.16%, respectively. No sequences of S. mansoni were detected in the cat blood samples or mouse blood samples. The PCR results were consistent with the mNGS results, confirming the accuracy of the mNGS analysis. In addition, during the detection process, the assembly-based analysis did not detect sequences of S. mansoni in all samples. In contrast, the read-based analysis successfully detected the target sequences without fail. Finally, the analysis of microbiota diversity in the definitive host faecal samples revealed that compared with those in the control group, the elevated microbial taxa in the infected group mainly were probiotics, such as Prevotella copri and Bifidobacterium adolescentis. Conversely, the decreased microbial populations were primarily associated with certain diseases, such as Collinsella stercoris and Catenibacterium sp. In this study, the diagnostic value of mNGS for S. mansoni infection was systematically evaluated. These findings establish a foundation for the more precise application of mNGS technology in the detection of S. mansoni and related cestode infections.
新一代宏基因组测序（mNGS）技术在寄生虫检测方面具有显著优势；然而，我们对其对曼氏螺虫感染的诊断价值仍知之甚少。在本研究中，采用mNGS技术分析了感染mansoni的猫的粪便样本和血液样本，以及感染mansoni S. plerocercotes幼虫的小鼠的组织样本和血液样本。此外，采用聚合酶链反应（PCR）对mNGS结果进行验证。系统评价mNGS对曼氏链球菌感染的诊断价值。mNGS结果显示，猫粪（CF）样品中mansoni S. reads计数分别为301497 （CF1）、1330549 （CF2）、1181162 （CF3）和0 (CF0)，相对丰度分别为3.17%、16.64%、13.14%和0%。在小鼠组织（MT）样品中，mansoni S. mansoni的reads计数分别为10791 （MT1）、438 （MT2）、3697 （MT3）和10 (MT0)，相对丰度分别为67.21%、3.65%、21.12%和0.16%。猫血和鼠血均未检出曼氏链球菌序列。PCR结果与mNGS结果一致，证实了mNGS分析的准确性。此外，在检测过程中，基于装配的分析并未在所有样品中检测到S. mansoni的序列。相比之下，基于读取的分析成功地检测到目标序列，没有失败。最后，对最终宿主粪便样本的微生物群多样性分析显示，与对照组相比，感染组微生物群的增加主要是益生菌，如copri普雷沃氏菌和青少年双歧杆菌。相反，微生物数量的减少主要与某些疾病有关，如粪Collinsella stercoris和Catenibacterium sp.。本研究系统评价了mNGS对mansoni s.m ansoni感染的诊断价值。这些发现为更精确地应用mNGS技术检测曼氏梭菌及相关寄生虫感染奠定了基础。
{"title":"Evaluation of the diagnostic value of metagenomic next-generation sequencing for zoonotic cestode Spirometra mansoni infection","authors":"Si Si Ru , Wen Li , Jie Hao , Cheng Yue Cao , Li Ma , Xi Zhang","doi":"10.1016/j.fawpar.2026.e00316","DOIUrl":"10.1016/j.fawpar.2026.e00316","url":null,"abstract":"<div><div>Metagenomic next-generation sequencing (mNGS) technology offers substantial advantages in parasite detection; however, we still know very little about its diagnostic value for <em>Spirometra mansoni</em> infection. In this study, mNGS technology was used to analyse faecal samples and blood samples from cats infected with <em>S. mansoni</em>, as well as tissue samples and blood samples from mice infected with the plerocercoid larvae of <em>S. mansoni</em>. Moreover, polymerase chain reaction (PCR) was employed to validate the mNGS results. The diagnostic value of mNGS for <em>S. mansoni</em> infection was systematically evaluated. The mNGS results revealed that the read counts of <em>S. mansoni</em> in the cat faeces (CF) samples were 301,497 (CF1), 1,330,549 (CF2), 1,181,162 (CF3), and 0 (CF0), with relative abundances of 3.17%, 16.64%, 13.14%, and 0%, respectively. In the mouse tissue (MT) samples, the read counts of <em>S. mansoni</em> were 10,791 (MT1), 438 (MT2), 3697 (MT3), and 10 (MT0), with relative abundances of 67.21%, 3.65%, 21.12%, and 0.16%, respectively. No sequences of <em>S. mansoni</em> were detected in the cat blood samples or mouse blood samples. The PCR results were consistent with the mNGS results, confirming the accuracy of the mNGS analysis. In addition, during the detection process, the assembly-based analysis did not detect sequences of <em>S. mansoni</em> in all samples. In contrast, the read-based analysis successfully detected the target sequences without fail. Finally, the analysis of microbiota diversity in the definitive host faecal samples revealed that compared with those in the control group, the elevated microbial taxa in the infected group mainly were probiotics, such as <em>Prevotella copri</em> and <em>Bifidobacterium adolescentis</em>. Conversely, the decreased microbial populations were primarily associated with certain diseases, such as <em>Collinsella stercoris</em> and <em>Catenibacterium</em> sp. In this study, the diagnostic value of mNGS for <em>S. mansoni</em> infection was systematically evaluated. These findings establish a foundation for the more precise application of mNGS technology in the detection of <em>S. mansoni</em> and related cestode infections.</div></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"42 ","pages":"Article e00316"},"PeriodicalIF":3.1,"publicationDate":"2026-01-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145927154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-01DOI: 10.1016/j.fawpar.2025.e00314
Ainun Nahar , Md. Farhan Hasan , Anas Bin Harun , Abdullah Al Bayazid , Tania Sultana , Jinnat Rehena , Joynti Saha , S.H.M. Faruk Siddiki , Md. Mizanur Rahman , Md. Ataur Rahman , Md Robiul Karim
Cryptosporidium spp. and Giardia duodenalis are intestinal protozoan parasites of zoonotic concern that cause gastrointestinal diseases in humans and various animals, including cats and dogs. This study investigates the prevalence, risk factors, and genetic diversity of Cryptosporidium spp. and G. duodenalis in domestic cats and dogs in Bangladesh to assess their zoonotic potential. Fecal samples were collected from 197 cats and 120 dogs in Dhaka and Gazipur metropolitan areas. We performed nested PCR targeting the small subunit ribosomal RNA (SSU rRNA) gene for Cryptosporidium and the β-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) genes for G. duodenalis, followed by nucleotide sequencing and analysis. The overall prevalence of Cryptosporidium was 8.1 % in cats and 4.2 % in dogs, whereas G. duodenalis was more common, detected in 29.9 % of cats and 25 % of dogs. Among examined variables, only sex and food types were significantly associated with G. duodenalis infection in dogs and cats, respectively. Molecular analysis identified three Cryptosporidium spp. in cats, including C. felis (81.3 %), C. baileyi (12.5 %), and C. canis (6.3 %), whereas 100 % of Cryptosporidium isolates from dogs were identified as C. canis. Multilocus genotyping of G. duodenalis revealed both host-adapted and zoonotic assemblages. Assemblage A predominated in both cats and dogs, followed by F and C in cats and C and D in dogs, with mixed infections observed in both hosts. The detection of zoonotic species and assemblages underscores the potential role of cats and dogs as reservoirs for human infection. These findings highlight the importance of monitoring intestinal protozoa in companion animals and promoting appropriate hygiene practices within a One Health framework.
{"title":"Molecular detection and zoonotic potential of Cryptosporidium spp. and Giardia duodenalis in cats and dogs from metropolitan areas of Bangladesh","authors":"Ainun Nahar , Md. Farhan Hasan , Anas Bin Harun , Abdullah Al Bayazid , Tania Sultana , Jinnat Rehena , Joynti Saha , S.H.M. Faruk Siddiki , Md. Mizanur Rahman , Md. Ataur Rahman , Md Robiul Karim","doi":"10.1016/j.fawpar.2025.e00314","DOIUrl":"10.1016/j.fawpar.2025.e00314","url":null,"abstract":"<div><div><em>Cryptosporidium</em> spp. and <em>Giardia duodenalis</em> are intestinal protozoan parasites of zoonotic concern that cause gastrointestinal diseases in humans and various animals, including cats and dogs. This study investigates the prevalence, risk factors, and genetic diversity of <em>Cryptosporidium</em> spp. and <em>G. duodenalis</em> in domestic cats and dogs in Bangladesh to assess their zoonotic potential. Fecal samples were collected from 197 cats and 120 dogs in Dhaka and Gazipur metropolitan areas. We performed nested PCR targeting the small subunit ribosomal RNA (<em>SSU rRNA</em>) gene for <em>Cryptosporidium</em> and the β-giardin (<em>bg</em>), glutamate dehydrogenase (<em>gdh</em>), and triosephosphate isomerase (<em>tpi</em>) genes for <em>G. duodenalis</em>, followed by nucleotide sequencing and analysis. The overall prevalence of <em>Cryptosporidium</em> was 8.1 % in cats and 4.2 % in dogs, whereas <em>G. duodenalis</em> was more common, detected in 29.9 % of cats and 25 % of dogs. Among examined variables, only sex and food types were significantly associated with <em>G. duodenalis</em> infection in dogs and cats, respectively. Molecular analysis identified three <em>Cryptosporidium</em> spp. in cats, including <em>C. felis</em> (81.3 %), <em>C. baileyi</em> (12.5 %), and <em>C. canis</em> (6.3 %), whereas 100 % of <em>Cryptosporidium</em> isolates from dogs were identified as <em>C. canis</em>. Multilocus genotyping of <em>G. duodenalis</em> revealed both host-adapted and zoonotic assemblages. Assemblage A predominated in both cats and dogs, followed by F and C in cats and C and D in dogs, with mixed infections observed in both hosts. The detection of zoonotic species and assemblages underscores the potential role of cats and dogs as reservoirs for human infection. These findings highlight the importance of monitoring intestinal protozoa in companion animals and promoting appropriate hygiene practices within a One Health framework.</div></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"42 ","pages":"Article e00314"},"PeriodicalIF":3.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145885124","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-20DOI: 10.1016/j.fawpar.2025.e00313
Cecilia Wangari Wambui , Mila Viaene , Hannah Njiriku Mwangi , Benjamin André , David Were Oguttu , Casim Umba Tolo , Bart Hellemans , Tine Huyse , Hugo F. Gante
Schistosomiasis, caused by parasites of the genus Schistosoma, remains a major public health burden in sub-Saharan Africa, particularly where access to clean water, sanitation, and hygiene is limited. Effective control requires large-scale surveillance, but traditional methods such as malacological surveys, and stool or urine microscopy often lack sensitivity and scalability. This study evaluated environmental DNA-based detection of Schistosoma mansoni in water samples from Lake Albert and Lake Victoria, Uganda. Three filtration techniques (open membrane, Waterra eDNA capsule, and Sylphium eDNA Dual filter capsule), were compared for eDNA yield and detection sensitivity. Quantitative PCR (qPCR) targeting the cytochrome c oxidase subunit 1 (COI) mitochondrial gene was used to quantify S. mansoni eDNA, following in silico and in vitro primer optimisation. Conventional malacological surveys were conducted in parallel for validation. Statistical analyses further examined associations between eDNA yield, detectability, and environmental factors. The qPCR assay had a practical limit of detection (LOD) of 100 DNA copies per reaction and a theoretical LOD/limit of quantification of 83 copies. Schistosoma mansoni eDNA was detected in 26 % (15/58) of samples from Lake Albert and 24 % (27/113) from Lake Victoria. Waterra filters yielded the most eDNA, and Sylphium purification produced significantly greater yields than column-based extraction kits. Both filter type and eDNA yield significantly influenced S. mansoni detection: Waterra and Sylphium-single filters had the highest amplification probabilities (∼40 %), while open membrane filters performed poorly (∼3 %). eDNA yield was a strong predictor of detection, with the odds of positivity increasing by ∼0.8 % per additional nanogram of eDNA. Among positive samples, Waterra filters produced the lowest mean Ct values, indicating greater recovery of amplifiable parasite DNA. Conversely, open membrane filters were the most affect by field contamination. Our findings highlight eDNA as a sensitive and scalable tool for surveillance of schistosomiasis and other water-borne parasitic diseases. While higher-capacity filters and two-phase extraction methods maximised eDNA yield, lower-yield methods still enabled detection in high-transmission settings. A comparative analysis of sampling effort, costs and contamination and infection risks is presented. Overall, our results support the adaptability of eDNA approaches across resource contexts and underscore the need for protocol standardisation, ecological validation, and field-deployable diagnostics such as LAMP.
由血吸虫属寄生虫引起的血吸虫病仍然是撒哈拉以南非洲的一个主要公共卫生负担，特别是在获得清洁水、环境卫生和个人卫生有限的地方。有效的控制需要大规模的监测，但传统的方法，如malacology调查，粪便或尿液显微镜往往缺乏灵敏度和可扩展性。本研究对乌干达艾伯特湖和维多利亚湖水样中的曼氏血吸虫环境dna检测方法进行了评价。比较了三种过滤技术（开放膜、Waterra eDNA胶囊和sylum eDNA双重过滤胶囊）的eDNA产率和检测灵敏度。采用针对细胞色素c氧化酶亚基1 （COI）线粒体基因的定量PCR （qPCR）方法，对mansoni S. mansoni的eDNA进行了定量分析，并进行了体外引物优化。为了验证，平行进行了常规的线虫学调查。统计分析进一步检验了eDNA产率、可检测性和环境因素之间的关系。qPCR检测的实际检测限（LOD）为每个反应100个DNA拷贝，理论定量限为83个拷贝。艾伯特湖和维多利亚湖样品中分别检出26%（15/58）和24%（27/113）的曼氏血吸虫eDNA。Waterra过滤器产生最多的eDNA，而sylium纯化产生的产量明显高于柱式提取试剂盒。滤器类型和eDNA产率都显著影响mansoni的检测：Waterra和sylm -single滤器的扩增概率最高（~ 40%），而开放式膜滤器的扩增概率较低（~ 3%）。eDNA产率是检测的一个强有力的预测指标，每增加一纳克eDNA，阳性几率增加约0.8%。在阳性样本中，Waterra过滤器产生的平均Ct值最低，表明可扩增寄生虫DNA的回收率更高。相反，开放式膜过滤器受现场污染的影响最大。我们的发现强调了eDNA作为血吸虫病和其他水传播寄生虫病监测的敏感和可扩展的工具。虽然高容量过滤器和两相提取方法可以最大限度地提高eDNA产量，但低产量方法仍然可以在高传输环境下进行检测。提出了取样努力、成本、污染和感染风险的比较分析。总的来说，我们的研究结果支持了eDNA方法在资源环境中的适应性，并强调了协议标准化、生态验证和现场可部署诊断（如LAMP）的必要性。
{"title":"Advancing schistosomiasis monitoring through optimised environmental DNA detection","authors":"Cecilia Wangari Wambui , Mila Viaene , Hannah Njiriku Mwangi , Benjamin André , David Were Oguttu , Casim Umba Tolo , Bart Hellemans , Tine Huyse , Hugo F. Gante","doi":"10.1016/j.fawpar.2025.e00313","DOIUrl":"10.1016/j.fawpar.2025.e00313","url":null,"abstract":"<div><div>Schistosomiasis, caused by parasites of the genus <em>Schistosoma</em>, remains a major public health burden in sub-Saharan Africa, particularly where access to clean water, sanitation, and hygiene is limited. Effective control requires large-scale surveillance, but traditional methods such as malacological surveys, and stool or urine microscopy often lack sensitivity and scalability. This study evaluated environmental DNA-based detection of <em>Schistosoma mansoni</em> in water samples from Lake Albert and Lake Victoria, Uganda. Three filtration techniques (open membrane, Waterra eDNA capsule, and Sylphium eDNA Dual filter capsule), were compared for eDNA yield and detection sensitivity. Quantitative PCR (qPCR) targeting the cytochrome <em>c</em> oxidase subunit 1 (COI) mitochondrial gene was used to quantify <em>S. mansoni</em> eDNA, following in silico and in vitro primer optimisation. Conventional malacological surveys were conducted in parallel for validation. Statistical analyses further examined associations between eDNA yield, detectability, and environmental factors. The qPCR assay had a practical limit of detection (LOD) of 100 DNA copies per reaction and a theoretical LOD/limit of quantification of 83 copies. <em>Schistosoma mansoni</em> eDNA was detected in 26 % (15/58) of samples from Lake Albert and 24 % (27/113) from Lake Victoria. Waterra filters yielded the most eDNA, and Sylphium purification produced significantly greater yields than column-based extraction kits. Both filter type and eDNA yield significantly influenced <em>S. mansoni</em> detection: Waterra and Sylphium-single filters had the highest amplification probabilities (∼40 %), while open membrane filters performed poorly (∼3 %). eDNA yield was a strong predictor of detection, with the odds of positivity increasing by ∼0.8 % per additional nanogram of eDNA. Among positive samples, Waterra filters produced the lowest mean Ct values, indicating greater recovery of amplifiable parasite DNA. Conversely, open membrane filters were the most affect by field contamination. Our findings highlight eDNA as a sensitive and scalable tool for surveillance of schistosomiasis and other water-borne parasitic diseases. While higher-capacity filters and two-phase extraction methods maximised eDNA yield, lower-yield methods still enabled detection in high-transmission settings. A comparative analysis of sampling effort, costs and contamination and infection risks is presented. Overall, our results support the adaptability of eDNA approaches across resource contexts and underscore the need for protocol standardisation, ecological validation, and field-deployable diagnostics such as LAMP.</div></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"42 ","pages":"Article e00313"},"PeriodicalIF":3.1,"publicationDate":"2025-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145841928","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-16DOI: 10.1016/j.fawpar.2025.e00312
Lavinia Ciuca , Maria Paola Maurelli , Antonio Bosco , Ines Hammami , Paola Vitiello , Mita Eva Sengupta , Anna-Sofie Stensgaard , Laura Rinaldi
This study investigated snails collected from eleven cattle farms in a Mediterranean area of southern Italy, where Fasciola hepatica (liver fluke) and Calicophoron daubneyi (rumen fluke) are known to occur. A total of 319 snails were collected from various aquatic habitats across the selected farms and identified using morphological and molecular analysis. BLAST analysis revealed two snail species: Galba truncatula (56.7 %) and Physella acuta (43.3 %). Statistical analyses revealed that shell and aperture lengths differed significantly between the two snail species. A subset of 130 snails was tested for the presence of F. hepatica and C. daubneyi DNA. Snails were initially tested in pools of ten individuals per species and single snails from positive pools were subsequently examined individually. Fasciola hepatica DNA was detected exclusively in G. truncatula, whereas C. daubneyi DNA was found in both G. truncatula and P. acuta. In addition, a total of 84 adult liver flukes were collected from cattle on seven of the eleven farms, morphometrically characterized, and molecularly confirmed as F. hepatica. The concurrent detection of fluke eggs in cattle faeces, adult flukes in livers and fluke DNA in snails suggests that active transmission is ongoing on these farms. Galba truncatula, already established as the main intermediate host for both F. hepatica and C. daubneyi in Europe, was confirmed in this study as naturally infected with both flukes under Italian field conditions. Broader seasonal surveys are warranted to better define infection dynamics. In contrast, the detection of C. daubneyi DNA in P. acuta requires experimental confirmation of cercarial shedding and infectivity to the definitive host to determine its actual role in fluke transmission.
{"title":"New insights into trematode infections in cattle and their snail intermediate hosts in a Mediterranean area of Italy","authors":"Lavinia Ciuca , Maria Paola Maurelli , Antonio Bosco , Ines Hammami , Paola Vitiello , Mita Eva Sengupta , Anna-Sofie Stensgaard , Laura Rinaldi","doi":"10.1016/j.fawpar.2025.e00312","DOIUrl":"10.1016/j.fawpar.2025.e00312","url":null,"abstract":"<div><div>This study investigated snails collected from eleven cattle farms in a Mediterranean area of southern Italy, where <em>Fasciola hepatica</em> (liver fluke) and <em>Calicophoron daubneyi</em> (rumen fluke) are known to occur. A total of 319 snails were collected from various aquatic habitats across the selected farms and identified using morphological and molecular analysis. BLAST analysis revealed two snail species: <em>Galba truncatula</em> (56.7 %) and <em>Physella acuta</em> (43.3 %). Statistical analyses revealed that shell and aperture lengths differed significantly between the two snail species. A subset of 130 snails was tested for the presence of <em>F. hepatica</em> and <em>C. daubneyi</em> DNA. Snails were initially tested in pools of ten individuals per species and single snails from positive pools were subsequently examined individually. <em>Fasciola hepatica</em> DNA was detected exclusively in <em>G. truncatula</em>, whereas <em>C. daubneyi</em> DNA was found in both <em>G. truncatula</em> and <em>P. acuta</em>. In addition, a total of 84 adult liver flukes were collected from cattle on seven of the eleven farms, morphometrically characterized, and molecularly confirmed as <em>F. hepatica</em>. The concurrent detection of fluke eggs in cattle faeces, adult flukes in livers and fluke DNA in snails suggests that active transmission is ongoing on these farms. <em>Galba truncatula</em>, already established as the main intermediate host for both <em>F. hepatica</em> and <em>C. daubneyi</em> in Europe, was confirmed in this study as naturally infected with both flukes under Italian field conditions. Broader seasonal surveys are warranted to better define infection dynamics. In contrast, the detection of <em>C. daubneyi</em> DNA in <em>P. acuta</em> requires experimental confirmation of cercarial shedding and infectivity to the definitive host to determine its actual role in fluke transmission.</div></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"42 ","pages":"Article e00312"},"PeriodicalIF":3.1,"publicationDate":"2025-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145791941","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-11DOI: 10.1016/j.fawpar.2025.e00308
Daniel Berdejo , Paula Nieto , Mª. Jesús Gracia , Ignacio de Blas , Sara Remón , Regina Lázaro , Susana Bayarri
Pork is recognized as a major source of Toxoplasma gondii infection in humans. Although the potential association between seropositivity in white pigs and the presence of T. gondii in their meat has been investigated, corresponding information on the Iberian pig breed is still limited. In this study, we investigated the presence of T. gondii in Iberian pork and assessed its correlation with individual serological profiles to evaluate whether antibody titres can serve as indicators of meat contamination. We tested the sera of 238 Iberian pigs from three southwestern Spanish provinces (Badajoz, Cáceres, and Córdoba) using an indirect immunofluorescence assay (IFA), and analyzed matched diaphragm samples by quantitative PCR (qPCR) for T. gondii DNA detection. Serological analysis revealed an overall seropositivity rate of 46.22 %, with significant regional differences (p ≤ 0.050). Córdoba exhibited the highest seropositivity (57.89 %), followed by Cáceres (48.38 %) and Badajoz (35.71 %). Concurrently, T. gondii DNA was present in 14.29 % of the diaphragm samples, with parasite loads ranging from 78.56 to 219.09 parasites/g. A statistically significant correlation (p ≤ 0.001) was observed between grouped IFA titres (<1:20, 1:20–1:40, ≥1:80) and qPCR positivity in the corresponding diaphragm samples. Notably, the proportion of animals with titres ≥1:80 closely matched the rate of qPCR-positive meat samples. We concluded that this serological threshold can serve as an effective screening tool to discriminate animals that are at a higher risk of harboring the parasite, thereby improving food safety within the HACCP-based safety system at the slaughterhouse and in the meat industry.
{"title":"Occurrence of Toxoplasma gondii in Iberian pork and its association with pig seropositivity","authors":"Daniel Berdejo , Paula Nieto , Mª. Jesús Gracia , Ignacio de Blas , Sara Remón , Regina Lázaro , Susana Bayarri","doi":"10.1016/j.fawpar.2025.e00308","DOIUrl":"10.1016/j.fawpar.2025.e00308","url":null,"abstract":"<div><div>Pork is recognized as a major source of <em>Toxoplasma gondii</em> infection in humans. Although the potential association between seropositivity in white pigs and the presence of <em>T. gondii</em> in their meat has been investigated, corresponding information on the Iberian pig breed is still limited. In this study, we investigated the presence of <em>T. gondii</em> in Iberian pork and assessed its correlation with individual serological profiles to evaluate whether antibody titres can serve as indicators of meat contamination. We tested the sera of 238 Iberian pigs from three southwestern Spanish provinces (Badajoz, Cáceres, and Córdoba) using an indirect immunofluorescence assay (IFA), and analyzed matched diaphragm samples by quantitative PCR (qPCR) for <em>T. gondii</em> DNA detection. Serological analysis revealed an overall seropositivity rate of 46.22 %, with significant regional differences (<em>p</em> ≤ 0.050). Córdoba exhibited the highest seropositivity (57.89 %), followed by Cáceres (48.38 %) and Badajoz (35.71 %). Concurrently, <em>T. gondii</em> DNA was present in 14.29 % of the diaphragm samples, with parasite loads ranging from 78.56 to 219.09 parasites/g. A statistically significant correlation (<em>p</em> ≤ 0.001) was observed between grouped IFA titres (<1:20, 1:20–1:40, ≥1:80) and qPCR positivity in the corresponding diaphragm samples. Notably, the proportion of animals with titres ≥1:80 closely matched the rate of qPCR-positive meat samples. We concluded that this serological threshold can serve as an effective screening tool to discriminate animals that are at a higher risk of harboring the parasite, thereby improving food safety within the HACCP-based safety system at the slaughterhouse and in the meat industry.</div></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"42 ","pages":"Article e00308"},"PeriodicalIF":3.1,"publicationDate":"2025-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145760731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Blastocystis is a genetically diverse enteric protist commonly found in humans and a wide range of vertebrate hosts. Although its prevalence and subtype (ST) distribution have been extensively studied in terrestrial ecosystems, its occurrence in marine organisms remains less known. In this study, we performed amplicon-based next-generation sequencing (NGS) to investigate, for the first time, the presence of Blastocystis in loggerhead sea turtles (Caretta caretta) and to expand existing data on ST diversity in cetaceans, stranded along the Italian Mediterranean coast. A total of 97 faecal samples were collected from 69 individuals of loggerhead sea turtles and 28 cetaceans. Blastocystis was detected in 44 % of the samples by real-time PCR—specifically in 39 loggerhead sea turtles and 4 fin whales (Balaenoptera physalus)—and further characterized by NGS. Ten STs were identified in loggerhead sea turtles and six in fin whales, with mixed infections frequently observed, particularly in turtles. Among the 18 STs detected, several represented new host records for marine organisms. ST4 was the most prevalent, especially in loggerhead sea turtles from the Tyrrhenian coast, and it exhibited a high degree of intra-subtype genetic variation. Comparison of ST4 sequences from this study with those of terrestrial origin revealed a certain level of substructuring; however, the most common haplotypes were shared between marine and terrestrial sources, supporting the hypothesis of a terrestrial origin for the marine STs. These findings highlight the potential use of Blastocystis STs occurring in marine megafauna as ecological indicators of faecal pollution from terrestrial origin in coastal marine environment. Moreover, they underscore the importance of applying a One Health framework, supported by NGS technologies, to elucidate the transmission dynamics of Blastocystis STs among humans, terrestrial, and marine hosts.
{"title":"Multiple Blastocystis subtypes in Mediterranean marine turtles and cetaceans by amplicon-based NGS","authors":"Marialetizia Palomba , Veronica Rodriguez-Fernandez , Renato Aco-Alburqueque , Meryam Carrus , Federica Marcer , Erica Marchiori , Mario Santoro , Tiziana Castrignanò , Daniele Canestrelli , Simonetta Mattiucci","doi":"10.1016/j.fawpar.2025.e00307","DOIUrl":"10.1016/j.fawpar.2025.e00307","url":null,"abstract":"<div><div><em>Blastocystis</em> is a genetically diverse enteric protist commonly found in humans and a wide range of vertebrate hosts. Although its prevalence and subtype (ST) distribution have been extensively studied in terrestrial ecosystems, its occurrence in marine organisms remains less known. In this study, we performed amplicon-based next-generation sequencing (NGS) to investigate, for the first time, the presence of <em>Blastocystis</em> in loggerhead sea turtles (<em>Caretta caretta</em>) and to expand existing data on ST diversity in cetaceans, stranded along the Italian Mediterranean coast. A total of 97 faecal samples were collected from 69 individuals of loggerhead sea turtles and 28 cetaceans. <em>Blastocystis</em> was detected in 44 % of the samples by real-time PCR—specifically in 39 loggerhead sea turtles and 4 fin whales (<em>Balaenoptera physalus</em>)—and further characterized by NGS. Ten STs were identified in loggerhead sea turtles and six in fin whales, with mixed infections frequently observed, particularly in turtles. Among the 18 STs detected, several represented new host records for marine organisms. ST4 was the most prevalent, especially in loggerhead sea turtles from the Tyrrhenian coast, and it exhibited a high degree of intra-subtype genetic variation. Comparison of ST4 sequences from this study with those of terrestrial origin revealed a certain level of substructuring; however, the most common haplotypes were shared between marine and terrestrial sources, supporting the hypothesis of a terrestrial origin for the marine STs. These findings highlight the potential use of <em>Blastocystis</em> STs occurring in marine megafauna as ecological indicators of faecal pollution from terrestrial origin in coastal marine environment. Moreover, they underscore the importance of applying a One Health framework, supported by NGS technologies, to elucidate the transmission dynamics of <em>Blastocystis</em> STs among humans, terrestrial, and marine hosts.</div></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"41 ","pages":"Article e00307"},"PeriodicalIF":3.1,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145747645","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01DOI: 10.1016/j.fawpar.2025.e00304
Zhuo Lan , Xue Wang , Yuxi Zhang , Lu Zhou , Xue Bai , Xinhui Zhang , Haokun Zhang , Hongyu Qiu , Junfeng Gao , Guofeng Cheng , Chunren Wang
Clonorchiasis, caused by Clonorchis sinensis, is a significant public health issue in China and East Asia. However, understanding its pathological mechanisms underlying this disease remains limited. Here, we isolated and characterized C. sinensis extracellular vesicles (CsEVs) and evaluated their uptake by Kupffer cells (KCs) in vitro in immortalized KC (ImKC) line and in vivo in a model of C. sinensis infection, investigating genes with altered gene expression after treatment with CsEVs and during progression to liver fibrosis. CsEVs were successfully taken up by KCs to regulate gene expression. RNA-sequencing analysis identified 694 differentially expressed genes (DEGs), including upregulation of those encoding colony-stimulating factor 3 (CSF3), IL1b, and others. Further validation showed increased expressions of these genes in mice infected with C. sinensis. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses suggested that these DEGs were involved in pathways such as TNF signaling, and NF-κB signaling. During liver fibrosis progression, CSF3 expression positively correlated with alpha smooth muscle actin (α-SMA) levels in the liver, both of which were higher compared with negative controls. CSF3 inhibition caused a significant decrease in α-SMA expression. This study was the first to report differential gene expression changes in ImKCs stimulated by CsEVs, with CSF3, the most significantly upregulated gene, having a potential role in liver fibrosis. These findings provide important data for the pathology of clonorchiasis and could identify new targets for treatment.
{"title":"Transcriptomic profiling of Kupffer cells exposed to Clonorchis sinensis extracellular vesicles: Unraveling the role of CSF3 in hepatic fibrosis","authors":"Zhuo Lan , Xue Wang , Yuxi Zhang , Lu Zhou , Xue Bai , Xinhui Zhang , Haokun Zhang , Hongyu Qiu , Junfeng Gao , Guofeng Cheng , Chunren Wang","doi":"10.1016/j.fawpar.2025.e00304","DOIUrl":"10.1016/j.fawpar.2025.e00304","url":null,"abstract":"<div><div>Clonorchiasis, caused by <em>Clonorchis sinensis</em>, is a significant public health issue in China and East Asia. However, understanding its pathological mechanisms underlying this disease remains limited. Here, we isolated and characterized <em>C. sinensis</em> extracellular vesicles (CsEVs) and evaluated their uptake by Kupffer cells (KCs) <em>in vitro</em> in immortalized KC (ImKC) line and <em>in vivo</em> in a model of <em>C. sinensis</em> infection, investigating genes with altered gene expression after treatment with CsEVs and during progression to liver fibrosis. CsEVs were successfully taken up by KCs to regulate gene expression. RNA-sequencing analysis identified 694 differentially expressed genes (DEGs), including upregulation of those encoding <em>colony-stimulating factor</em> 3 (<em>CSF</em>3), <em>IL1b</em>, and others. Further validation showed increased expressions of these genes in mice infected with <em>C. sinensis</em>. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses suggested that these DEGs were involved in pathways such as TNF signaling, and NF-κB signaling. During liver fibrosis progression, <em>CSF</em>3 expression positively correlated with alpha smooth muscle actin (α-SMA) levels in the liver, both of which were higher compared with negative controls. <em>CSF</em>3 inhibition caused a significant decrease in α-SMA expression. This study was the first to report differential gene expression changes in ImKCs stimulated by CsEVs, with <em>CSF</em>3, the most significantly upregulated gene, having a potential role in liver fibrosis. These findings provide important data for the pathology of clonorchiasis and could identify new targets for treatment.</div></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"41 ","pages":"Article e00304"},"PeriodicalIF":3.1,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145623731","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01DOI: 10.1016/j.fawpar.2025.e00305
P. Ramos , C. Ayra-Pardo , M.M. Oliveira , L.F. Rangel , F. Atroch , C. Sirin , M.J. Santos
The black scabbardfish, Aphanopus carbo, is a deep-sea Atlantic fish with socioeconomic value, but it is often heavily infected with Anisakis spp. larvae. The aim of the present work was to ascertain the distribution and quantification of Anisakis spp. larvae in the viscera and muscle of A. carbo, and to evaluate whether larval intensity in the viscera can be used to predict parasite load in the muscle. UV-press and pepsin-HCl digestion methods were complemented by molecular techniques to detect and identify the larvae. Four batches of four individuals each, one batch per season (n = 64), were obtained. Each batch, with a different parasite intensity based on visual inspection, was selected for comparison. Larval subsamples randomly selected from the viscera, belly flaps, and dorsal muscle were identified by PCR-RFLP, marking the first study of this kind in this species. Data showed that the higher the larval load in the visceral cavity and internal organs, the greater the number of larvae in the belly flaps, allowing us to predict Anisakis burden in this site. The low number and unpredictability of the zoonotic A. simplex s.s in the dorsal muscle pose a risk to consumers. Epidemiological data support discarding the belly flaps from fish with relatively high parasite intensity. Additional recommendations to mitigate anisakids presence in fishery products are presented for improvement and implementation along the black scabbardfish value chain.
{"title":"Anisakis spp. larvae in black scabbardfish (Aphanopus carbo) muscle: Consumer health risk and recommendations along the value chain","authors":"P. Ramos , C. Ayra-Pardo , M.M. Oliveira , L.F. Rangel , F. Atroch , C. Sirin , M.J. Santos","doi":"10.1016/j.fawpar.2025.e00305","DOIUrl":"10.1016/j.fawpar.2025.e00305","url":null,"abstract":"<div><div>The black scabbardfish, <em>Aphanopus carbo</em>, is a deep-sea Atlantic fish with socioeconomic value, but it is often heavily infected with <em>Anisakis</em> spp. larvae. The aim of the present work was to ascertain the distribution and quantification of <em>Anisakis</em> spp. larvae in the viscera and muscle of <em>A. carbo</em>, and to evaluate whether larval intensity in the viscera can be used to predict parasite load in the muscle. UV-press and pepsin-HCl digestion methods were complemented by molecular techniques to detect and identify the larvae. Four batches of four individuals each, one batch per season (<em>n</em> = 64), were obtained. Each batch, with a different parasite intensity based on visual inspection, was selected for comparison. Larval subsamples randomly selected from the viscera, belly flaps, and dorsal muscle were identified by PCR-RFLP, marking the first study of this kind in this species. Data showed that the higher the larval load in the visceral cavity and internal organs, the greater the number of larvae in the belly flaps, allowing us to predict <em>Anisakis</em> burden in this site. The low number and unpredictability of the zoonotic <em>A. simplex s.s</em> in the dorsal muscle pose a risk to consumers. Epidemiological data support discarding the belly flaps from fish with relatively high parasite intensity. Additional recommendations to mitigate anisakids presence in fishery products are presented for improvement and implementation along the black scabbardfish value chain.</div></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"41 ","pages":"Article e00305"},"PeriodicalIF":3.1,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145690793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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