Human gnathostomiasis is a food-borne zoonotic helminthic infection widely reported in Latin America, Asia, and Southeast Asia. Consuming raw, or under-cooked fresh-water fish is the leading cause of this helminthic infection, which is clinically characterized by signs of inflammation, itching sensation, or irritation with migratory swelling. Neurological symptoms resulting from neurognathostomiasis vary, and there is scant information due to the rareness of patient brain samples. This study aimed to demonstrate the first evidence of human neurognathostomiasis by the detection of Gnathostoma spinigerum larva in patient's brain during craniotomy, supported by histopathological, immunological and proteomic evidence. Clinical symptoms were obtained from medical history and physical examination with laboratory investigations, including magnetic resonance imaging (MRI), left temporal craniotomy, histopathology of brain tissue, and Western blot analysis, were performed to elucidate the causative pathogens for diagnosis. In addition, the host–parasite interaction of the parasite invading the patient's brain was characterized through proteomics. Histopathology revealed worms with the characteristic cuticular spines of G. spinigerum which were detected and identified. These histopathological findings were consistent with a positive Western blot showing a 24-kDa reactive-band for gnathostomiasis. Proteomic analysis revealed the presence of G. spinigerum serpin and serine protease in the patient's serum. Moreover, the leucine-rich alpha-2-glycoprotein was indicated as a systemic biomarker of early brain injury related to invasion by G. spinigerum. Therefore, our study provides the initial evidence of human neurognathostomiasis due to G. spinigerum larval invasion along with successful craniotomy and proven larval detection including complete follow-up, and the disease prognosis after surgical treatment.
人类钩端螺旋体病是一种食源性人畜共患病,在拉丁美洲、亚洲和东南亚有广泛报道。食用生的或未煮熟的淡水鱼是导致这种蠕虫感染的主要原因,临床表现为发炎、瘙痒或刺激症状,并伴有移行性肿胀。神经性寄生虫病导致的神经症状各不相同,由于患者脑部样本稀少,因此相关信息很少。本研究旨在通过组织病理学、免疫学和蛋白质组学证据,首次证明在开颅手术中在患者大脑中发现了棘尾蚋幼虫。通过病史和体格检查获得临床症状,并进行实验室检查,包括磁共振成像(MRI)、左颞部开颅手术、脑组织组织病理学检查和 Western 印迹分析,以明确诊断的致病病原体。此外,还通过蛋白质组学分析了侵入患者大脑的寄生虫的宿主-寄生虫相互作用特征。组织病理学发现,寄生虫具有刺毛虫特有的角质刺,并对其进行了检测和鉴定。这些组织病理学结果与阳性 Western 印迹显示的 24-kDa 钩端螺旋体反应带一致。蛋白质组分析表明,患者血清中含有刺吸虫血清素和丝氨酸蛋白酶。此外,富含亮氨酸的甲-2-糖蛋白被认为是与刺吸虫入侵有关的早期脑损伤的全身生物标志物。因此,我们的研究提供了因棘尾蝽幼虫入侵导致人类神经寄生虫病的初步证据,以及成功的开颅手术和经证实的幼虫检测,包括完整的随访和手术治疗后的疾病预后。
{"title":"Detection of Gnathostoma spinigerum larva in the brain with complete follow-up after surgical treatment of human neurognathostomiasis","authors":"Chayanuchit Chayangsu , Sumate Ampawong , Onrapak Reamtong , Parnpen Viriyavejakul , Tapanee Kanjanapruthipong , Kamonpan Fongsodsri , Suwaphat Intapun , Pongwat Polpong , Rattanarat Intarat , Prakaykaew Charunwatthana , Abigail Hui En Chan , Dorn Watthanakulpanich","doi":"10.1016/j.fawpar.2024.e00229","DOIUrl":"https://doi.org/10.1016/j.fawpar.2024.e00229","url":null,"abstract":"<div><p>Human gnathostomiasis is a food-borne zoonotic helminthic infection widely reported in Latin America, Asia, and Southeast Asia. Consuming raw, or under-cooked fresh-water fish is the leading cause of this helminthic infection, which is clinically characterized by signs of inflammation, itching sensation, or irritation with migratory swelling. Neurological symptoms resulting from neurognathostomiasis vary, and there is scant information due to the rareness of patient brain samples. This study aimed to demonstrate the first evidence of human neurognathostomiasis by the detection of <em>Gnathostoma spinigerum</em> larva in patient's brain during craniotomy, supported by histopathological, immunological and proteomic evidence. Clinical symptoms were obtained from medical history and physical examination with laboratory investigations, including magnetic resonance imaging (MRI), left temporal craniotomy, histopathology of brain tissue, and Western blot analysis, were performed to elucidate the causative pathogens for diagnosis. In addition, the host–parasite interaction of the parasite invading the patient's brain was characterized through proteomics. Histopathology revealed worms with the characteristic cuticular spines of <em>G. spinigerum</em> which were detected and identified. These histopathological findings were consistent with a positive Western blot showing a 24-kDa reactive-band for gnathostomiasis. Proteomic analysis revealed the presence of <em>G. spinigerum</em> serpin and serine protease in the patient's serum. Moreover, the leucine-rich alpha-2-glycoprotein was indicated as a systemic biomarker of early brain injury related to invasion by <em>G. spinigerum.</em> Therefore, our study provides the initial evidence of human neurognathostomiasis due to <em>G. spinigerum</em> larval invasion along with successful craniotomy and proven larval detection including complete follow-up, and the disease prognosis after surgical treatment.</p></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"35 ","pages":"Article e00229"},"PeriodicalIF":2.5,"publicationDate":"2024-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405676624000118/pdfft?md5=fcf7652d053b0e92153302481a3da542&pid=1-s2.0-S2405676624000118-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140947155","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Enterocytozoon bieneusi is one of the most prevalent microsporidia species, responsible for more than 90% of human and animal microsporidiosis. Microsporidia species, particularly E. bieneusi, are frequently reported from waterborne and foodborne outbreaks. Therefore, early detection is crucial in clinics and outbreak investigations. This study aimed to design a loop-mediated isothermal amplification (LAMP) for rapid detection of E. bieneusi. Total DNA was extracted from 30 E. bieneusi –positive samples, which had been confirmed with nested PCR. LAMP primers were designed based on the identical fragment of small subunit ribosomal RNA (SSU rRNA) gene. LAMP reactions were performed at 63 °C for 60 min. The sensitivity and specificity of the assay were analyzed and the results of amplification were compared to real-time PCR. Our results showed that the LAMP assay successfully amplified 25/30 (83.3%) samples. The specificity results indicated no false positive with other microorganisms. Furthermore, the LAMP method exhibited a sensitivity (limit of detection, LoD) as low as 34 ag/μL of total DNA. Compared to the LAMP assay, real-time PCR was able to detect all 30 nested PCR-positive samples. Our findings showed that the LAMP assay was able to detect 83.3% of E. bieneusi-positive samples. Although the current assay was not able to detect all nested PCR-positive samples, the lack of need for specific instruments, rapid processes, and high specificity makes LAMP assay a suitable tool for screening.
{"title":"Design and evaluation of loop-mediated isothermal amplification for rapid detection of Enterocytozoon bieneusi","authors":"Fatemeh Mahdavi , Hamed Mirjalali , Maryam Niyyati , Seyyed Javad Seyyed Tabaei , Amir Shamloo , Hamid Asadzadeh Aghdaei","doi":"10.1016/j.fawpar.2024.e00225","DOIUrl":"https://doi.org/10.1016/j.fawpar.2024.e00225","url":null,"abstract":"<div><p><em>Enterocytozoon bieneusi</em> is one of the most prevalent microsporidia species, responsible for more than 90% of human and animal microsporidiosis. Microsporidia species, particularly <em>E. bieneusi,</em> are frequently reported from waterborne and foodborne outbreaks. Therefore, early detection is crucial in clinics and outbreak investigations. This study aimed to design a loop-mediated isothermal amplification (LAMP) for rapid detection of <em>E. bieneusi.</em> Total DNA was extracted from 30 <em>E. bieneusi</em> –positive samples, which had been confirmed with nested PCR. LAMP primers were designed based on the identical fragment of small subunit ribosomal RNA (<em>SSU rRNA</em>) gene. LAMP reactions were performed at 63 °C for 60 min. The sensitivity and specificity of the assay were analyzed and the results of amplification were compared to real-time PCR. Our results showed that the LAMP assay successfully amplified 25/30 (83.3%) samples. The specificity results indicated no false positive with other microorganisms. Furthermore, the LAMP method exhibited a sensitivity (limit of detection, LoD) as low as 34 ag/μL of total DNA. Compared to the LAMP assay, real-time PCR was able to detect all 30 nested PCR-positive samples. Our findings showed that the LAMP assay was able to detect 83.3% of <em>E. bieneusi-</em>positive samples. Although the current assay was not able to detect all nested PCR-positive samples, the lack of need for specific instruments, rapid processes, and high specificity makes LAMP assay a suitable tool for screening.</p></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"35 ","pages":"Article e00225"},"PeriodicalIF":2.5,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405676624000076/pdfft?md5=b0e8a221af6d8b2c0a4de899ce0e7bc5&pid=1-s2.0-S2405676624000076-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140138187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-26DOI: 10.1016/j.fawpar.2024.e00222
Maria Francesca Sini , Maria Manconi , Antonio Varcasia , Giovanna Massei , Ramona Sandu , Naunain Mehmood , Fahad Ahmed , Carlo Carta , Cinzia Cantacessi , Christian Scarano , Antonio Scala , Claudia Tamponi
Toxoplasma gondii is a zoonotic parasite able of infecting all warm-blooded animals. Toxoplasmosis is one of the major foodborne diseases globally. The consumption of wild boar (Sus scrofa) meat from recreational hunting has been linked to outbreaks of human toxoplasmosis. The island of Sardinia (Italy) contains a large wild boar population, thus providing an opportunity to assess the distribution of Toxoplasma in this species and the associated risks of transmission to humans. A total of 562 wild boars were screened: heart and meat juice samples were tested for T. gondii DNA via nested-PCR and IgG anti-Toxoplasma by commercial ELISA. Anti-Toxoplasma IgG were detected in 24.6% (138/562) of animals, while 37.2% (209/562) of the heart samples were PCR positive. The prevalence of T. gondii antibodies and DNA highlights the potential role of wild boar as an important reservoir for this parasite. The study suggests that wild boar could play a significant role in spreading the parasite to humans. As wild boar numbers are increasing throughout their range, their potential role in transmitting toxoplasmosis should be communicated to stakeholders, and the impact of different population control methods on disease transmission should be thoroughly assessed to mitigate potential threats effectively.
{"title":"Seroepidemiological and biomolecular survey on Toxoplasma gondii in Sardinian wild boar (Sus scrofa)","authors":"Maria Francesca Sini , Maria Manconi , Antonio Varcasia , Giovanna Massei , Ramona Sandu , Naunain Mehmood , Fahad Ahmed , Carlo Carta , Cinzia Cantacessi , Christian Scarano , Antonio Scala , Claudia Tamponi","doi":"10.1016/j.fawpar.2024.e00222","DOIUrl":"10.1016/j.fawpar.2024.e00222","url":null,"abstract":"<div><p><em>Toxoplasma gondii</em> is a zoonotic parasite able of infecting all warm-blooded animals. Toxoplasmosis is one of the major foodborne diseases globally. The consumption of wild boar (<em>Sus scrofa</em>) meat from recreational hunting has been linked to outbreaks of human toxoplasmosis. The island of Sardinia (Italy) contains a large wild boar population, thus providing an opportunity to assess the distribution of <em>Toxoplasma</em> in this species and the associated risks of transmission to humans. A total of 562 wild boars were screened: heart and meat juice samples were tested for <em>T. gondii</em> DNA via nested-PCR and IgG anti-<em>Toxoplasma</em> by commercial ELISA. Anti-<em>Toxoplasma</em> IgG were detected in 24.6% (138/562) of animals, while 37.2% (209/562) of the heart samples were PCR positive. The prevalence of <em>T. gondii</em> antibodies and DNA highlights the potential role of wild boar as an important reservoir for this parasite. The study suggests that wild boar could play a significant role in spreading the parasite to humans. As wild boar numbers are increasing throughout their range, their potential role in transmitting toxoplasmosis should be communicated to stakeholders, and the impact of different population control methods on disease transmission should be thoroughly assessed to mitigate potential threats effectively.</p></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"34 ","pages":"Article e00222"},"PeriodicalIF":2.5,"publicationDate":"2024-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405676624000040/pdfft?md5=a967dae9850c0babb09bc56f9c833778&pid=1-s2.0-S2405676624000040-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139634133","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-26DOI: 10.1016/j.fawpar.2024.e00223
Azdayanti Muslim , Shafiq Aazmi , Yi Xian Er , Shezryna Shahrizal , Yvonne Ai Lian Lim
In indigenous populations where soil-transmitted helminths (STH) infections are endemic, STH parasites (i.e., Ascaris lumbricoides, Trichuris trichiura, hookworms) often co-exist and co-evolve with the gut microbiota of their human hosts. The association between STH infections and the gut microbiota of the colonized human hosts has been established, but few studies explored the gut microbiota of the parasites. This preliminary study aimed to characterize the microbiota of the STH parasite for further understanding the STH parasite-host relationship. The gut microbial genomic DNA from four adult A. lumbricoides worms recovered from a six-year-old indigenous Negrito boy living in an STH-endemic village in Perak, Peninsular Malaysia was extracted and sequenced for the V3-V4 region of the 16S rRNA. The microbiota profiles of these worms were characterized and compared with the gut microbiota of their human host, including the profiles from four STH-positive and three STH-negative individuals from the same tribe and village. The gut microbial structure of A. lumbricoides was found to be differed significantly from their human host. The worms contained lower gut bacterial abundance and diversity than human. This difference was evident in the beta diversity analysis which showed a clear separation between the two sample types. While both Firmicutes (52.3%) and Bacteroidetes (36.6%) are the predominant phyla followed by Proteobacteria (7.2%) in the human gut, the microbiota of Ascaris gut is highly dominated by Firmicutes, constituting 84.2% relative abundance (mainly from the genus Clostridium), followed by Proteobacteria (11.1%), Tenericutes (1.8%) and Bacteroidetes (1.5%). The parasites were also found to alter the microbial structure of the human gut following infection based on the relatively higher bacterial abundance in STH-positive versus STH-negative participants. Further studies with a greater number of Ascaris adults and human hosts are needed to confirm the gut microbiota profiles.
{"title":"Ascaris lumbricoides harbors a distinct gut microbiota profile from its human host: Preliminary insights","authors":"Azdayanti Muslim , Shafiq Aazmi , Yi Xian Er , Shezryna Shahrizal , Yvonne Ai Lian Lim","doi":"10.1016/j.fawpar.2024.e00223","DOIUrl":"10.1016/j.fawpar.2024.e00223","url":null,"abstract":"<div><p>In indigenous populations where soil-transmitted helminths (STH) infections are endemic, STH parasites (i.e., <em>Ascaris lumbricoides</em>, <em>Trichuris trichiura</em>, hookworms) often co-exist and co-evolve with the gut microbiota of their human hosts. The association between STH infections and the gut microbiota of the colonized human hosts has been established, but few studies explored the gut microbiota of the parasites. This preliminary study aimed to characterize the microbiota of the STH parasite for further understanding the STH parasite-host relationship. The gut microbial genomic DNA from four adult <em>A. lumbricoides</em> worms recovered from a six-year-old indigenous Negrito boy living in an STH-endemic village in Perak, Peninsular Malaysia was extracted and sequenced for the V3-V4 region of the 16S rRNA. The microbiota profiles of these worms were characterized and compared with the gut microbiota of their human host, including the profiles from four STH-positive and three STH-negative individuals from the same tribe and village. The gut microbial structure of <em>A. lumbricoides</em> was found to be differed significantly from their human host. The worms contained lower gut bacterial abundance and diversity than human. This difference was evident in the beta diversity analysis which showed a clear separation between the two sample types. While both Firmicutes (52.3%) and Bacteroidetes (36.6%) are the predominant phyla followed by Proteobacteria (7.2%) in the human gut, the microbiota of <em>Ascaris</em> gut is highly dominated by Firmicutes, constituting 84.2% relative abundance (mainly from the genus <em>Clostridium</em>), followed by Proteobacteria (11.1%), Tenericutes (1.8%) and Bacteroidetes (1.5%). The parasites were also found to alter the microbial structure of the human gut following infection based on the relatively higher bacterial abundance in STH-positive versus STH-negative participants. Further studies with a greater number of <em>Ascaris</em> adults and human hosts are needed to confirm the gut microbiota profiles.</p></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"34 ","pages":"Article e00223"},"PeriodicalIF":2.5,"publicationDate":"2024-01-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405676624000052/pdfft?md5=2735f290487fed1829432dd4d110a9b3&pid=1-s2.0-S2405676624000052-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139634341","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-21DOI: 10.1016/j.fawpar.2024.e00221
Oleg V. Shcherbakov , Sargis A. Aghayan , Hasmik Sh. Gevorgyan , Tigran A. Abgaryan , Ruzanna H. Gevorgyan , Alejandro Jiménez-Meléndez , Lucy J. Robertson
The intestinal protozoan parasites, Cryptosporidium and Giardia, are known to have a global distribution, infecting and causing disease in a range of hosts, including people, livestock, pets, and wildlife. However, data from some regions is very sparse. In Armenia, in the Caucasus region of West Asia, only scanty data are available, with just a few surveys on Cryptosporidium infections in livestock, and no available data on human infections or environmental contamination. As part of implementation of water analysis methods for these parasites in Armenia, 24 raw water samples and two sediment samples were analysed for these parasites using a range of approaches, including modified Ziehl-Neelsen, Lugol stain, immunofluorescent antibody test (IFAT), qPCR and, on sediment samples, immunomagnetic separation and IFAT. Results suggest substantial contamination of raw water sources and indicate the need for further targeted studies using appropriate methods and collecting data on host infections in catchment areas.
{"title":"Preliminary investigations of parasite contamination of water sources in Armenia","authors":"Oleg V. Shcherbakov , Sargis A. Aghayan , Hasmik Sh. Gevorgyan , Tigran A. Abgaryan , Ruzanna H. Gevorgyan , Alejandro Jiménez-Meléndez , Lucy J. Robertson","doi":"10.1016/j.fawpar.2024.e00221","DOIUrl":"10.1016/j.fawpar.2024.e00221","url":null,"abstract":"<div><p>The intestinal protozoan parasites, <em>Cryptosporidium</em> and <em>Giardia</em>, are known to have a global distribution, infecting and causing disease in a range of hosts, including people, livestock, pets, and wildlife. However, data from some regions is very sparse. In Armenia, in the Caucasus region of West Asia, only scanty data are available, with just a few surveys on <em>Cryptosporidium</em> infections in livestock, and no available data on human infections or environmental contamination. As part of implementation of water analysis methods for these parasites in Armenia, 24 raw water samples and two sediment samples were analysed for these parasites using a range of approaches, including modified Ziehl-Neelsen, Lugol stain, immunofluorescent antibody test (IFAT), qPCR and, on sediment samples, immunomagnetic separation and IFAT. Results suggest substantial contamination of raw water sources and indicate the need for further targeted studies using appropriate methods and collecting data on host infections in catchment areas.</p></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"34 ","pages":"Article e00221"},"PeriodicalIF":2.5,"publicationDate":"2024-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405676624000039/pdfft?md5=ff125e65ff5c5057ab01e767ed73396d&pid=1-s2.0-S2405676624000039-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139632252","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-01-19DOI: 10.1016/j.fawpar.2024.e00220
Selene Rubiola , Gastón Moré , Tiziana Civera , Andrew Hemphill , Caroline F. Frey , Walter Basso , Irene Colasanto , Davide Vercellino , Marta Fidelio , Mauro Lovisone , Francesco Chiesa
<div><p>Bovine eosinophilic myositis is an inflammatory myopathy characterized by multiple focal or diffuse grey to green patches leading to condemnation of affected carcasses. Although its etiology is still uncertain, there is evidence that <em>Sarcocystis</em> species may play a role in the development of eosinophilic myositis. The goal of the present study was to identify <em>Sarcocystis</em> spp. in intralesional and extralesional tissues of condemned cattle carcasses, in order to evaluate the possible role of different bovine <em>Sarcocystis</em> spp. in the etiology of bovine eosinophilic myositis. Muscle samples (<em>n</em> = 100) of 26 affected carcasses were collected in Northern Italy. One to five samples with lesions and two aliquots of tissue without lesions were collected from each carcass; lesions were grossly categorized in green focal lesions and green diffuse patches. Genomic DNA was extracted and analyzed by multiplex-PCR targeting different <em>Sarcocystis</em> spp. Unidentified species were characterized morphologically (light microscopy, histology), ultrastructurally (scanning and transmission electron microscopy) and on the molecular level (complete 18S rRNA gene and partial <em>cox1</em> gene sequencing). A bovine eosinophilic myositis prevalence of 0.017% was visually assessed by routine carcass inspection between 2014 and 2019 in Italy (184/1,108,150 slaughtered cattle). Out of 26 carcasses, 25 revealed the presence of at least one <em>Sarcocystis</em> species (96.2%). The presence of <em>Sarcocystis</em> spp. DNA was significantly more frequent in intralesional than in extralesional samples. Considering the different species, <em>Sarcocystis bovifelis</em> and <em>Sarcocystis hominis</em> were significantly more frequent in intralesional (41.7% and 50%, respectively) than in extralesional samples (1.9% and 15.4%, respectively), while there was no significant difference between the presence of <em>Sarcocystis cruzi</em> and <em>Sarcocystis hirsuta</em> in intralesional (27.1% and 2.1%, respectively) and extralesional (30.8% and 1.9%, respectively) samples. The presence of an unnamed <em>Sarcocystis</em> sp. showing thick-walled (3.7–5.4 μm) cysts with densely packed, flattened, undulating and narrow protrusions, which showed an S-shape in side view, was recorded in the diaphragm of two carcasses. Genomic DNA from individual sarcocysts isolated from the diaphragm was successfully amplified and further sequenced. Sequence comparison revealed <94.6% and 83.4% identity at 18S rRNA and <em>cox1</em> genes, respectively, with other named <em>Sarcocystis</em> spp., while the phylogenetic analysis clearly separated the unnamed <em>Sarcocystis</em> sp. from the other <em>Sarcocystis</em> spp. using cattle as intermediate hosts. The present study contributes to the understanding of the importance of different <em>Sarcocystis</em> spp. in the pathogenesis of bovine eosinophilic myositis. The results emphasize the association of <
{"title":"Detection of Sarcocystis hominis, Sarcocystis bovifelis, Sarcocystis cruzi, Sarcocystis hirsuta and Sarcocystis sigmoideus sp. nov. in carcasses affected by bovine eosinophilic myositis","authors":"Selene Rubiola , Gastón Moré , Tiziana Civera , Andrew Hemphill , Caroline F. Frey , Walter Basso , Irene Colasanto , Davide Vercellino , Marta Fidelio , Mauro Lovisone , Francesco Chiesa","doi":"10.1016/j.fawpar.2024.e00220","DOIUrl":"https://doi.org/10.1016/j.fawpar.2024.e00220","url":null,"abstract":"<div><p>Bovine eosinophilic myositis is an inflammatory myopathy characterized by multiple focal or diffuse grey to green patches leading to condemnation of affected carcasses. Although its etiology is still uncertain, there is evidence that <em>Sarcocystis</em> species may play a role in the development of eosinophilic myositis. The goal of the present study was to identify <em>Sarcocystis</em> spp. in intralesional and extralesional tissues of condemned cattle carcasses, in order to evaluate the possible role of different bovine <em>Sarcocystis</em> spp. in the etiology of bovine eosinophilic myositis. Muscle samples (<em>n</em> = 100) of 26 affected carcasses were collected in Northern Italy. One to five samples with lesions and two aliquots of tissue without lesions were collected from each carcass; lesions were grossly categorized in green focal lesions and green diffuse patches. Genomic DNA was extracted and analyzed by multiplex-PCR targeting different <em>Sarcocystis</em> spp. Unidentified species were characterized morphologically (light microscopy, histology), ultrastructurally (scanning and transmission electron microscopy) and on the molecular level (complete 18S rRNA gene and partial <em>cox1</em> gene sequencing). A bovine eosinophilic myositis prevalence of 0.017% was visually assessed by routine carcass inspection between 2014 and 2019 in Italy (184/1,108,150 slaughtered cattle). Out of 26 carcasses, 25 revealed the presence of at least one <em>Sarcocystis</em> species (96.2%). The presence of <em>Sarcocystis</em> spp. DNA was significantly more frequent in intralesional than in extralesional samples. Considering the different species, <em>Sarcocystis bovifelis</em> and <em>Sarcocystis hominis</em> were significantly more frequent in intralesional (41.7% and 50%, respectively) than in extralesional samples (1.9% and 15.4%, respectively), while there was no significant difference between the presence of <em>Sarcocystis cruzi</em> and <em>Sarcocystis hirsuta</em> in intralesional (27.1% and 2.1%, respectively) and extralesional (30.8% and 1.9%, respectively) samples. The presence of an unnamed <em>Sarcocystis</em> sp. showing thick-walled (3.7–5.4 μm) cysts with densely packed, flattened, undulating and narrow protrusions, which showed an S-shape in side view, was recorded in the diaphragm of two carcasses. Genomic DNA from individual sarcocysts isolated from the diaphragm was successfully amplified and further sequenced. Sequence comparison revealed <94.6% and 83.4% identity at 18S rRNA and <em>cox1</em> genes, respectively, with other named <em>Sarcocystis</em> spp., while the phylogenetic analysis clearly separated the unnamed <em>Sarcocystis</em> sp. from the other <em>Sarcocystis</em> spp. using cattle as intermediate hosts. The present study contributes to the understanding of the importance of different <em>Sarcocystis</em> spp. in the pathogenesis of bovine eosinophilic myositis. The results emphasize the association of <","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"34 ","pages":"Article e00220"},"PeriodicalIF":2.5,"publicationDate":"2024-01-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405676624000027/pdfft?md5=19a87d4e20e31fa3411277713f06a6a2&pid=1-s2.0-S2405676624000027-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139549106","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alveolar echinococcosis is a zoonotic disease caused by a larval-stage Echinococcus multilocularis infection. Geographical haplotyping targeting the parasite's mitochondrial cytochrome b (cob) gene has been reported for isolates from definitive and intermediate hosts (wild canids and rodents); however, there are limited reports on strain typing for the dead-end host, the horse, which could act as a sentinel for E. multilocularis. Accordingly, we investigated the diversity of E. multilocularis in isolates obtained from slaughtered Japanese and Canadian horses originating from the Iburi and Hidaka regions in Hokkaido and from Alberta, respectively, with PCR and haplogroup analyses targeting cob gene sequences obtained. Seventy horses were diagnosed with alveolar echinococcosis based on histopathology and cob-gene PCR testing. The E. multilocularis detected in these horses was classified as either an Asian (for Hokkaido-raised horses) or a European (for Alberta-raised horses) haplogroup, based on the obtained cob-gene sequence analysis. In addition, haplotype network analysis revealed that E. multilocularis isolated from Hokkaido-raised horses is highly homologous to Kazakhstan isolates, and E. multilocularis isolated from Alberta-raised horses is highly homologous to Austrian isolates. The results of this study suggest that cob-gene-targeted PCR analysis could be useful for the geographical genetic characterization of E. multilocularis isolated from horses.
{"title":"Molecular phylogenetic analysis of Echinococcus multilocularis from horses raised in Canada or Japan, using mitochondrial cytochrome b gene–targeted PCR","authors":"Tatsuro Hifumi , Tetsuya Tanaka , Ichinosuke Suzu , Miho Sato , Kohei Akioka , Chiaki Fujimata , Ryohei Shinkai , Tsutomu Maeda , Kodai Kusakisako , Hiromi Ikadai , Noriaki Miyoshi","doi":"10.1016/j.fawpar.2024.e00219","DOIUrl":"https://doi.org/10.1016/j.fawpar.2024.e00219","url":null,"abstract":"<div><p>Alveolar echinococcosis is a zoonotic disease caused by a larval-stage <em>Echinococcus multilocularis</em> infection. Geographical haplotyping targeting the parasite's mitochondrial cytochrome <em>b</em> (<em>cob</em>) gene has been reported for isolates from definitive and intermediate hosts (wild canids and rodents); however, there are limited reports on strain typing for the dead-end host, the horse, which could act as a sentinel for E<em>. multilocularis</em>. Accordingly, we investigated the diversity of <em>E. multilocularis</em> in isolates obtained from slaughtered Japanese and Canadian horses originating from the Iburi and Hidaka regions in Hokkaido and from Alberta, respectively, with PCR and haplogroup analyses targeting <em>cob</em> gene sequences obtained. Seventy horses were diagnosed with alveolar echinococcosis based on histopathology and <em>cob</em>-gene PCR testing. The <em>E. multilocularis</em> detected in these horses was classified as either an Asian (for Hokkaido-raised horses) or a European (for Alberta-raised horses) haplogroup, based on the obtained <em>cob</em>-gene sequence analysis. In addition, haplotype network analysis revealed that <em>E. multilocularis</em> isolated from Hokkaido-raised horses is highly homologous to Kazakhstan isolates, and <em>E. multilocularis</em> isolated from Alberta-raised horses is highly homologous to Austrian isolates. The results of this study suggest that <em>cob</em>-gene-targeted PCR analysis could be useful for the geographical genetic characterization of <em>E. multilocularis</em> isolated from horses.</p></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"34 ","pages":"Article e00219"},"PeriodicalIF":2.5,"publicationDate":"2024-01-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405676624000015/pdfft?md5=737cbd62f9e959c8ccc698f5e2d0de53&pid=1-s2.0-S2405676624000015-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139480174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-12-09DOI: 10.1016/j.fawpar.2023.e00214
P. Suarez , A. Vallejos-Almirall , I. Fernández , I. Gonzalez-Chavarria , J.L. Alonso , G. Vidal
Cryptosporidium parvum and Blastocystis hominis are foodborne parasites known for causing diarrhea. They accumulate in mussels grown on contaminated water bodies, due to the discharge of treated sewage from sewage treatment plants (STP). Despite this, some countries like Chile do not include these parasites in the control or monitoring of sewage water. The objective of this research was to evaluate the contamination of C. parvum. and B. hominis from treated sewage (disinfected by chlorination) and Cholga mussels in a touristic rural cove from the bay of Concepción. Cholga mussels from commercial stores and a treated sewage sample were analyzed. Cryptosporidium spp. was identified by Ziehl-Neelsen-Staining (ZNS) and C. parvum by direct-immunofluorescence assay (IFA) from ZNS-positive samples. Blastocystis hominis was identified by PCR using locus SSU rDNA. C. parvum and B. hominis subtype ST3 were found in 40% and 45% of Cholga mussel samples, respectively, and both parasites were identified in the treated sewage. Blastocystis hominis SSU rDNA gene alignment from Cholga mussels and treated sewage showed 89% of similarity, indicating that could be the same parasite in both samples. We describe the first evidence of possible contamination with these parasites from treated sewage to Cholga mussel suggesting an environmental contamination with high human risk. Based on these results, further studies will consider all the rural coves and STP from the bay to prevent possible contamination of these parasites.
{"title":"Identification of Cryptosporidium parvum and Blastocystis hominis subtype ST3 in Cholga mussel and treated sewage: Preliminary evidence of fecal contamination in harvesting area","authors":"P. Suarez , A. Vallejos-Almirall , I. Fernández , I. Gonzalez-Chavarria , J.L. Alonso , G. Vidal","doi":"10.1016/j.fawpar.2023.e00214","DOIUrl":"https://doi.org/10.1016/j.fawpar.2023.e00214","url":null,"abstract":"<div><p><em>Cryptosporidium parvum</em> and <em>Blastocystis hominis</em> are foodborne parasites known for causing diarrhea. They accumulate in mussels grown on contaminated water bodies, due to the discharge of treated sewage from sewage treatment plants (STP). Despite this, some countries like Chile do not include these parasites in the control or monitoring of sewage water. The objective of this research was to evaluate the contamination of <em>C. parvum</em>. and <em>B. hominis</em> from treated sewage (disinfected by chlorination) and Cholga mussels in a touristic rural cove from the bay of Concepción. Cholga mussels from commercial stores and a treated sewage sample were analyzed. <em>Cryptosporidium</em> spp. was identified by Ziehl-Neelsen-Staining (ZNS) and <em>C. parvum</em> by direct-immunofluorescence assay (IFA) from ZNS-positive samples. <em>Blastocystis hominis</em> was identified by PCR using locus SSU rDNA. <em>C. parvum</em> and <em>B. hominis</em> subtype ST3 were found in 40% and 45% of Cholga mussel samples, respectively, and both parasites were identified in the treated sewage. <em>Blastocystis hominis</em> SSU rDNA gene alignment from Cholga mussels and treated sewage showed 89% of similarity, indicating that could be the same parasite in both samples. We describe the first evidence of possible contamination with these parasites from treated sewage to Cholga mussel suggesting an environmental contamination with high human risk. Based on these results, further studies will consider all the rural coves and STP from the bay to prevent possible contamination of these parasites.</p></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"34 ","pages":"Article e00214"},"PeriodicalIF":2.5,"publicationDate":"2023-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405676623000276/pdfft?md5=f45512db938b8382cafd6ab22d608415&pid=1-s2.0-S2405676623000276-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138838855","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}