Faecal prevalence and histopathological evaluation of coccidiosis in bovine calves

Faecal prevalence and histopathological evaluation of coccidiosis in bovine calves | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Faecal prevalence and histopathological evaluation of coccidiosis in bovine calves Vikas Jaiswal, Bhupinder singh sandhu, Apminderpal singh brar, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2456431/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 24 May, 2023 Read the published version in Journal of Parasitic Diseases → Version 1 posted 4 You are reading this latest preprint version Abstract Bovine coccidiois, caused by Eimeria spp. is widely prevalent around the globe and responsible for huge economic losses by causing morbidity and mortality among young calves.The present study was designed to evaluate the prevalence as well as to evaluate histopathological alteration associated with it. The faecal samples were collected from 700 bovine calves uptotwo month of age from August 2019 to July 2021 and screened for Eimeria oocycts. The intestinal tissue samples of 37 calves were also collected which died during the study period after showing symptoms of diarrhea and examined for histological lesions.The faecal prevalence of Eimeria observed in our study was 2.29% (16/700) while in tissue samples only two out of 37 were foundpositive for Eimeria infection. Tissue sections reveled various stages of Eimeria gametogony, variable congestion, haemorrhage, and necrosis along with cryptic dilatation and mononuclear cell infiltration.Coccidia was not found to be associated with season, age and sex of calf. Bovine coccidiosiswas found to be endemic with low prevalence but severe onset characterized by moderate to severe congestion and inflammatory reaction mainly in the ileum and caecum. Eimeria Prevalence histopathology Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Introduction Eimeria is an obligate, intracellular,monoxenous, protozoan parasites of the phylum Apicomplexa and family Eimeriidae. It is one of the most important parasitoses reported in bovines around the globe and responsible for enormous economic losses in dairy industry (Li et al., 2021 ). E. bovis E. zurenii and E. bareillyi are three most common and most pathogenicspecies(Mundtet al., 2005 ;Manyaet al., 2008 ;Ramirez et al. 2009 ;Das et al. 2015 ;Nain et al. 2017 ;Pyziel et al. 2020 ). It mainly exerts its pathological effect by causing damage to intestinal mucosa of large and small intestineduring its sexual reproduction. The infection occurs through faeco-oral rout by ingestion of sporulated oocysts which are very resistant and remain viable in the environment for months in adverse climatic conditions (Cruvinel et al. 2018 ). Several risk factors such as seasonality, humidity, age, sex, change in feed and management systemetc. were reported by previous researchers (Girma 2017; Das et al. 2015 ). There is paucity of reports especially in regards to histopathological evaluation of naturally infected Eimeria calves in India, so the current study was planned not only to estimate the prevalence but also to investigate the histopathological lesions in naturally infected bovine calves. Material And Methods A total 700 faecal samples were collected from bovine calves from August 2019 to July 2021. The samples were collected from organized and unorganized farms in and around Ludhiana district of Punjab which is located between 30° − 33’, 31° -1’ north latitudes and 75° -25’ and 76° -27’ east longitudes, elevation ranges from about 216 meters in the west to about 268 meters in the east. It lies in humid subtropical climate with three distinct seasons; summer, winter and monsoon. Faecal smears were screened for oocysts after staining with Leishman and modified ZiehlNeelsen stain as per standard protocol. During the study period thirty seven calves were found dead after showing symptoms of diarrhea. Post mortem examination were carried out and gross lesions were recorded. The intestinal tissue samples were collected andfixed in 10% neutral buffered formalin. Paraffin embedded tissue sections (4 micron) were stained with hematoxylin and eosin stain by standard procedure (Luna 1968).The H&E stained slides were examined under microscope (Olympus BX53) and microphotography was carried out by using DP27 Olympus camera and cellsenssoftware. Results The clinical signs like diarrhea, dysentery, dehydration, lethargy, inanition, emaciation, rough hair coat, anemia of varying severity was observed in most of the cases. In the current study, only 2.29% (16) faecal samples were found positive for coccidian oocyst out of 700 samples. Two species were identified on the basis of morphometric examination (Rehmanet al. 2011 ) consisting of E. bovis (Fig. 5 ) and E.subspherica (Fig. 6 ). Mean oocyst size of E. bovis measured in our study was20.24µm X28.29µm while E.subspherica was 10.78µm X12.22µm. We also detected three cases of mixed infection with Cryptosporidium (Fig. 7 ). Prevalence and results of Chi square test in relation to season, age and sex have been shown in the Table 1 and 2 respectively. We observed highest prevalence of Eimeria in winter (2.80%) followed by summer (2.40%), monsoon (2.38%) and post monsoon (1.60%) season. Sex wise prevalence was higher in male calves (2.72%) than female calves (1.80%).Age wise prevalence was highest in age group IV (12.31%) followed by age group V(12.00%), age group III (5.91%), age group II (0.32%) and age group I (0.00%). However, Chi square test has shown non-significant association with all these three variables (Table 2 ). Gross lesions of intestinal tissue samples observed in current study were congestion and haemorrhage in different parts of small and large intestine with highest severity in ileum and caecum. The mucosa was oedematous thickened, and covered with necrotic debris material. Ballooning of intestine with gas and diarrheic faeces was evident. Microscopic examination revealed moderate to high congestion of mucosa and submucosa, goblet cell hyperplasia(Fig. 3 ), infiltration of inflammatory mononuclear cells in lamina propria and Lymphoid cell depletion in payers patches (Fig. 1 ). There was variable necrosis of mucosa and accumulation of debris material in the lumen(Fig. 4 ). Destruction of villi led to stunting and alteration in crypt–villi ratio (Fig. 1 ). In addition, mucosal haemorrhages and disruption of muscularis mucosae were also seen at places. Different stages of parasitic gametogony were distinctly visible (Fig. 2 ) in ileum and caecum. The parasite not only confined to mucosa but also invaded the submucosa and nodules of parasitic aggregation (coccidial nest) were seen in some of the Payer's patches (Fig. 1 ). Both male and female gamonts were detected in the same mucosal crypts of affected area. Sexually differentiated macrogamonts (female gamonts) were identified on the basis of centrally placed nucleus surrounded by large eosinophilic amylopectin granules (wall form bodies) while microgamonts (male gamonts) withnumerous nuclei (Fig. 2 ).Histology of mesenteric lymph node reveled lymhadenitis with infiltration of neutrophils and macropahges at places in one case while varying depletion of lymphoid cells and congestion in both the cases. Discussion In the current study, we observed only 2.27% (16) Eimeria prevalence which was in line with the finding of Brar et al. ( 2017 ) and Hussain et al. ( 2017 ) who reported only 4 and 2.9% prevalence respectively. In contrast to our finding, most of the researchers around the globe reported much higher prevalence ranging from 6.0%to 91.7% (Manya et al. 2008 ;Das et al. 2015 ;Girma 2017;Nain et al. 2017 ;Morgoglione et al. 2020 ; Li et al. 2021 ). The reason for getting very low prevalence in our study may be the age of the calf from which samples were collected and prepatient period of parasite (Mundt et al. 2005 ; Li et al. 2021 ). In our study we collected 87.14% samples from less than three week of age group animals. We did not find any positive case in first week of age and only one case in second week. Another reason may be easy availability of effective drugs against this parasite in this area and increased awareness about hygienic practices. Li et al. ( 2021 ) did meta-analysis of online available data of coccidial prevalence in China upto February 2020 and concluded reducing trend of prevalence year after year (from 57.9% before 2000 to 25.0% after 2015). Morgoglione et al.(2020) reported higher prevalence both in summer (82.4% and winter (82.3%)) season in buffalo calveswhile Das et al ( 2015 ) reported highest prevalence of E. bovis in pre monsoon (62.8%), followed by monsoon (58.6%), winter (55.4%) and summer season(50.8%).Higher prevalence observed in male calves in current study is in agreement with most of the previous researchers (Nain et al. 2017 ;Heidari et al. 2014 ).Though we found higher prevalence in older group of animals and male sex but statistically (Chi square test) it is non-significant. Direct life cycle of this parasite (without any intermediate host) and zoonotic implication explains the non-significant association of the cryptosporidium with above the risk variables. Similar to our study Mundt et al. ( 2005 ) also reported watery to haemorrhagic diarrhea but they observed lesions starting from terminal jejunum upto mid colon in experimentally infected calves. We observed highest severity of gross lesions in ileum and caecum. In contrast, Pyziel et al. ( 2020 ) recorded gross lesions mainly in large intestine. Hussain et al. ( 2017 ) noted specific transverse zebra striping on the mucosal surface in all the case along with haemorrhage and necrosis. White raised nodules have been reported in small ruminants (Satish et al. 2019 ) but non apparent in bovine coccidiosis. Variable pathology of different species of Eimeria has been reported by various scientists (Das et al. 2015 ;Cruvinel et al. 2018 ). Our histological findings are in line with Hussain et al. ( 2017 ) who also documented mononuclear cellular infiltration in the lamina propria, desquamation of epithelial lining, and severe hemorrhages with mucosal scars and coccidial cells nested in the lamina propria in all infected animals. Mundt et al. ( 2005 ) reportedmicroscopic lesions starting from initial mucosal hyperemia to acute typhlitis and necrotising colitis. They also observed infiltration of granulocytic series and proliferation of goblet cell. However, Sultana et al. ( 2015 ) reported highest severity both in caecum and colon. They observed mild hyperemia along with granulocytic mucosal infiltration at 15 day post infection (dpi) and acute sever lesion (degeneration, shortened and necrosis of villi, haemorrhages in mucosa and sub-mucosa, widening of crypts of lieburkuhn and oedema of lamina propria) at 25dpi. Stockdale ( 1977 ) also reported similar findings along with fibrin deposits and diphtheritic membrane on mucosa at 18dpi followed by resolution and mucosal regeneration commencing from 22nd to 32nd dpi but they observed lesions mainly in large intestine.Kawahara ( 2016 ) also observed similar lesion produced by gametogony both in small and large intestine. The lesions observed in our study such as severe congestion destruction of villus epithelium was mainly attributed to gametogony which can be seen in Fig. 1 by presence of different developmental stages of Eimeria deep into submucosa. As a result of gametogony there was massive destruction and stunting of villi. In an effort of regeneration, there was proliferation of crypt epithelium. Oedema of mucosa appeared due damage of villus epithelium as well as leaky blood vessels of lamina propria. The absorptive mature villus epithelium was replaced by immature crypt epithelium with secretory activity which causes diarrhea and when there is destruction of villi, it leads to haemorrhage and dysentery. The invasion of Eimeria to submucosalpayers patches of current study have not been reported by previous researches. Most of the scientists reported mucosal involvement only (Mundt et al. 2005 ;Sultana 2015;Hussain et al. ( 2017 ). Conclusion Bovine coccidiosis is an endemic with low prevalence but with severe onset of disease.Histology revealed its predilection of distal small intestine and proximal large intestine. The gross pathology revealed lesions varying from mild congestion to haemorrhagic enteritis, necrosis, ballooning and enlargement of mesenteric lymph node. Microscopically, lesions include congestion, mucosal oedema, mononuclear cell infiltration, destruction and stunting of villus epithelium. Parasitic gametogony responsible for villus destruction, cryptic dilatation, accumulation of necrotic debris and replacement of mature villus epithelium with immature secretory cryptic epithelium leading to malabsorption, diarrhea, dehydration and death. Coccidial prevalence was not found to be associated with age, sex and season. Declarations Acknowledgement: The authors acknowledge the Science & Engineering Research Board (SERB), Department of Science & Technology, Government of India for providing funds through project entitled “Biomolecular detection of cryptosporidium and associated etiological agents from faecal samples of diarrheic bovine calves” project no. EMR/2017/000540/AS Funding: Science & Engineering Research Board (SERB), Department of Science & Technology, Government of India, Project no. EMR/2017/000540/AS Compliance with ethical standards: The field study was approved by the Institutional ethical committee - IAEC/2019/188-221. Competing Interests: The authors of this research article declare that they have no conflict of interest Undertaking : The manuscript has not been published previously in full form or partially, and is not under consideration for publication elsewhere. Authors contribution: 1. Dr. Vikas Jaiswal: Sample collection, processing and manuscript writing 2. Dr.Apminder Pal Singh Brar: Designed the project, execution, monitoring and statistical analysis (project principal investigator) 3. Dr.Bhupinder Singh Sandhu: Photomicrography, micrometry and histopathological evaluation 4. Lachhman Das Singla: Parasitic identification and manuscript editing References Brar A, Sood NK, Kaur P, Singla LD, Sandhu BS, Gupta K, Narang D, Singh CK, Chandra M (2017) Periurban outbreaks of bovine calf scours in Northern India caused by Cryptosporidium in association with other enteropathogens. Epidemiol Infect 145(13):2717–2726. https://doi.org/10.1017/S0950268817001224 Cruvinel LB, Nicaretta JE, Bastos T, Couto L, Santos J, Zapa D, Cavalcante A, Cruz BC, BorgesD, Borges FA, Soares VE, Lopes W (2018) Eimeria species in dairy and beef cattle of different ages in Goiás state, Brazil. 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Variable No of samples No of positive samples Prevalence (%) Season Winter 178 5 2.80 Summer 125 3 2.40 Monsoon 210 5 2.38 Post monsoon 187 3 1.60 Age group Group I (0–7 day) 221 0 0.00 Group II (8–14 day) 312 1 0.32 Group III (15–21 day) 77 4 5.19 Group IV (22–30 day) 65 8 12.31 Group V (31–60 day) 25 3 12.00 Sex Male 368 10 2.72 Female 332 6 1.80 Total 700 16 2.29 Table 2 Significant relationship of variables (season, age and aex) with Eimeria spp. infection in faecal samples of calves. Etiological agent Season Age Sex Chi-Square p-Value Chi-Square p-Value Chi-Square p-Value Eimeria 8.533 0.036 4.668 0.458 0.528 0.467 P < 0.01 considered significant. Cite Share Download PDF Status: Published Journal Publication published 24 May, 2023 Read the published version in Journal of Parasitic Diseases → Version 1 posted Reviewers agreed at journal 16 Jan, 2023 Reviewers invited by journal 16 Jan, 2023 Editor assigned by journal 09 Jan, 2023 First submitted to journal 08 Jan, 2023 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2456431","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":167960753,"identity":"c35c1623-69a1-4311-826b-a09d4cee5be4","order_by":0,"name":"Vikas Jaiswal","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABEklEQVRIiWNgGAWjYBACxgbGBiBlwcDGw8DA/MNAQg4keuABYS0SEC0MFTbGYC0JhC2TYGAAazmTlggygQGfFub2w82feWokEvt4Dj/+XNh2OH1+2OGHQFvs5HQbcDisJ7FNmueYRGIbb5uZ9My2w7kbb6cZALUkG5sdwOWXxDbmHDYJYzZ+BjMGXpCW2QkgLQcSt+HS0v+w+XPOP5AW9s8fgFrSDWenf8CvZUZig3Rum4QcG2+PgTTPmbQEeekcArbMeNgm/bcPqIXnTJnkjAobww3SOQUHEgxw+8WwP/3xxxnfbHjke9I3f/hgICEvPxvEqLCTw6mlAV3EAKzSALtyEJDHFMEwZBSMglEwCkY6AAADeWFaWH7nlgAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0001-5457-3419","institution":"Sardar Vallabh Bhai Patel University of Agriculture and Technology","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Vikas","middleName":"","lastName":"Jaiswal","suffix":""},{"id":167960754,"identity":"97b1930c-3c7f-496e-bd9d-8e0e0bb8724b","order_by":1,"name":"Bhupinder singh sandhu","email":"","orcid":"","institution":"Guru Angad Dev Veterinary and Animal Sciences University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Bhupinder","middleName":"singh","lastName":"sandhu","suffix":""},{"id":167960755,"identity":"dd568889-9464-44c2-8d60-4e09136015d0","order_by":2,"name":"Apminderpal singh brar","email":"","orcid":"","institution":"Guru Angad Dev Veterinary and Animal Sciences University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Apminderpal","middleName":"singh","lastName":"brar","suffix":""},{"id":167960756,"identity":"0a8298fe-1652-4353-a68b-5c554a740e01","order_by":3,"name":"Lachhman Das Singla","email":"","orcid":"","institution":"Guru Angad Dev Veterinary and Animal Sciences University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Lachhman","middleName":"Das","lastName":"Singla","suffix":""}],"badges":[],"createdAt":"2023-01-08 17:45:25","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2456431/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2456431/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s12639-023-01590-x","type":"published","date":"2023-05-24T20:58:54+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":31710578,"identity":"8ad58515-6969-440f-ba53-cf28878483cf","added_by":"auto","created_at":"2023-01-17 21:17:04","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":2053373,"visible":true,"origin":"","legend":"\u003cp\u003ePhotomicrograph of ileum showing coccidia infection in mucosa \u0026amp;coccidial nest in submucosal payers patches (Blue arrow), depletion of lymphoid cell in payers patches (yellow arrow), congestion of mucosal \u0026amp;submucosal blood vessels (red arrow) and disruption of muscularis mucosae (green arrow), destruction and stunting (red arrow head) of villi (Stain H\u0026amp;E; Scale bar 200 µm).\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-2456431/v1/6b7c7cea93e0c17cb78fdea1.jpeg"},{"id":31710584,"identity":"862ba729-af66-4544-ba2e-4506e68db72b","added_by":"auto","created_at":"2023-01-17 21:17:05","extension":"jpeg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1578618,"visible":true,"origin":"","legend":"\u003cp\u003ePhotomicrograph showing various developmental stages of coccidia. Macrogamonts (red arrow) with large eosinophilic amylopectin granules (wall form bodies), microgamonts (blue arrow) possessed numerous nuclei and mature oocysts (black arrow) (Stain H\u0026amp;E; Scale bar 50 µm).\u003c/p\u003e","description":"","filename":"floatimage2.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-2456431/v1/00b25015bc80856a0c21e055.jpeg"},{"id":31711338,"identity":"c935e9ba-5f7d-4cb3-a8af-88424f9a7295","added_by":"auto","created_at":"2023-01-17 21:25:05","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":2618258,"visible":true,"origin":"","legend":"\u003cp\u003ePhotomicrograph showing goblet cell hyperplasia and mononuclear infiltration of mucosa (Stain H\u0026amp;E; Scale bar 50 µm).\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-2456431/v1/5ee6b39edfa37d7a08748f07.png"},{"id":31710582,"identity":"09e58674-7e20-460a-b43c-8f9840de10b3","added_by":"auto","created_at":"2023-01-17 21:17:05","extension":"jpeg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":824255,"visible":true,"origin":"","legend":"\u003cp\u003ePhotomicrograph showing marked accumulation of necrotic debris in the intestinal lumen, destruction of villi, congestion and infiltration of mononuclear inflammatory cells (Stain H\u0026amp;E; Scale bar 200 µm).\u003c/p\u003e","description":"","filename":"floatimage4.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-2456431/v1/7ce6aa12af1b4db038488563.jpeg"},{"id":31710579,"identity":"ac53fd2e-cdf6-4c58-bb8c-0bcefc52b3b9","added_by":"auto","created_at":"2023-01-17 21:17:05","extension":"jpeg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":108594,"visible":true,"origin":"","legend":"\u003cp\u003eFaecal smear showing \u003cem\u003eE. bovis\u003c/em\u003eoocysts in leishman stained smear (Scale bar 10 µm).\u003c/p\u003e","description":"","filename":"floatimage5.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-2456431/v1/592789966ee4d9f2bcde2783.jpeg"},{"id":31710581,"identity":"59840486-41c8-4b44-8dfb-571d731e6b32","added_by":"auto","created_at":"2023-01-17 21:17:05","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":929129,"visible":true,"origin":"","legend":"\u003cp\u003eFaecal smear showing \u003cem\u003eE. subsphherica\u003c/em\u003e oocysts in modified ZN stained smear (Scale bar 10 µm).\u003c/p\u003e","description":"","filename":"floatimage6.png","url":"https://assets-eu.researchsquare.com/files/rs-2456431/v1/0867aa3f06d97436c40fb15e.png"},{"id":31711337,"identity":"feca3938-9a4e-4438-a321-e6b3f9b04e57","added_by":"auto","created_at":"2023-01-17 21:25:05","extension":"jpeg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":75402,"visible":true,"origin":"","legend":"\u003cp\u003eFaecal smear showing mixed infection ofEimeria and Cryptosporidium spp. oocysts stained with modified ZN stain (Scale bar 20 µm).\u003c/p\u003e","description":"","filename":"floatimage7.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-2456431/v1/61d71633c85c1d67280ac2e7.jpeg"},{"id":44730974,"identity":"638be497-e1e8-4f3f-be03-d7dc2e80a28e","added_by":"auto","created_at":"2023-10-16 21:38:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":4466845,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2456431/v1/ee8648c9-e99c-423b-a2ea-e3855f375fae.pdf"}],"financialInterests":"","formattedTitle":"Faecal prevalence and histopathological evaluation of coccidiosis in bovine calves","fulltext":[{"header":"Introduction","content":"\u003cp\u003eEimeria is an obligate, intracellular,monoxenous, protozoan parasites of the phylum Apicomplexa and family Eimeriidae. It is one of the most important parasitoses reported in bovines around the globe and responsible for enormous economic losses in dairy industry (Li et al., \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). \u003cem\u003eE. bovis E. zurenii\u003c/em\u003e and \u003cem\u003eE. bareillyi are three\u003c/em\u003e most common and most pathogenicspecies(Mundtet al., \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2005\u003c/span\u003e;Manyaet al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2008\u003c/span\u003e;Ramirez et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2009\u003c/span\u003e;Das et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e;Nain et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2017\u003c/span\u003e;Pyziel et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). It mainly exerts its pathological effect by causing damage to intestinal mucosa of large and small intestineduring its sexual reproduction. The infection occurs through faeco-oral rout by ingestion of sporulated oocysts which are very resistant and remain viable in the environment for months in adverse climatic conditions (Cruvinel et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Several risk factors such as seasonality, humidity, age, sex, change in feed and management systemetc. were reported by previous researchers (Girma 2017; Das et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). There is paucity of reports especially in regards to histopathological evaluation of naturally infected Eimeria calves in India, so the current study was planned not only to estimate the prevalence but also to investigate the histopathological lesions in naturally infected bovine calves.\u003c/p\u003e"},{"header":"Material And Methods","content":"\u003cp\u003eA total 700 faecal samples were collected from bovine calves from August 2019 to July 2021. The samples were collected from organized and unorganized farms in and around Ludhiana district of Punjab which is located between 30\u0026deg; \u0026minus;\u0026thinsp;33\u0026rsquo;, 31\u0026deg; -1\u0026rsquo; north latitudes and 75\u0026deg; -25\u0026rsquo; and 76\u0026deg; -27\u0026rsquo; east longitudes, elevation ranges from about 216 meters in the west to about 268 meters in the east. It lies in humid subtropical climate with three distinct seasons; summer, winter and monsoon. Faecal smears were screened for oocysts after staining with Leishman and modified ZiehlNeelsen stain as per standard protocol. During the study period thirty seven calves were found dead after showing symptoms of diarrhea. Post mortem examination were carried out and gross lesions were recorded. The intestinal tissue samples were collected andfixed in 10% neutral buffered formalin. Paraffin embedded tissue sections (4 micron) were stained with hematoxylin and eosin stain by standard procedure (Luna 1968).The H\u0026amp;E stained slides were examined under microscope (Olympus BX53) and microphotography was carried out by using DP27 Olympus camera and cellsenssoftware.\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003eThe clinical signs like diarrhea, dysentery, dehydration, lethargy, inanition, emaciation, rough hair coat, anemia of varying severity was observed in most of the cases. In the current study, only 2.29% (16) faecal samples were found positive for coccidian oocyst out of 700 samples. Two species were identified on the basis of morphometric examination (Rehmanet al. \u003cspan class=\"CitationRef\"\u003e2011\u003c/span\u003e) consisting of \u003cem\u003eE. bovis\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e) and \u003cem\u003eE.subspherica\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e). Mean oocyst size of \u003cem\u003eE. bovis\u003c/em\u003emeasured in our study was20.24\u0026micro;m X28.29\u0026micro;m while \u003cem\u003eE.subspherica\u003c/em\u003e was 10.78\u0026micro;m X12.22\u0026micro;m. We also detected three cases of mixed infection with Cryptosporidium (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e). Prevalence and results of Chi square test in relation to season, age and sex have been shown in the Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e respectively. We observed highest prevalence of Eimeria in winter (2.80%) followed by summer (2.40%), monsoon (2.38%) and post monsoon (1.60%) season. Sex wise prevalence was higher in male calves (2.72%) than female calves (1.80%).Age wise prevalence was highest in age group IV (12.31%) followed by age group V(12.00%), age group III (5.91%), age group II (0.32%) and age group I (0.00%). However, Chi square test has shown non-significant association with all these three variables (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\n\u003cp\u003eGross lesions of intestinal tissue samples observed in current study were congestion and haemorrhage in different parts of small and large intestine with highest severity in ileum and caecum. The mucosa was oedematous thickened, and covered with necrotic debris material. Ballooning of intestine with gas and diarrheic faeces was evident. Microscopic examination revealed moderate to high congestion of mucosa and submucosa, goblet cell hyperplasia(Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e), infiltration of inflammatory mononuclear cells in lamina propria and Lymphoid cell depletion in payers patches (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). There was variable necrosis of mucosa and accumulation of debris material in the lumen(Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). Destruction of villi led to stunting and alteration in crypt\u0026ndash;villi ratio (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). In addition, mucosal haemorrhages and disruption of muscularis mucosae were also seen at places. Different stages of parasitic gametogony were distinctly visible (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e) in ileum and caecum. The parasite not only confined to mucosa but also invaded the submucosa and nodules of parasitic aggregation (coccidial nest) were seen in some of the Payer\u0026apos;s patches (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). Both male and female gamonts were detected in the same mucosal crypts of affected area. Sexually differentiated macrogamonts (female gamonts) were identified on the basis of centrally placed nucleus surrounded by large eosinophilic amylopectin granules (wall form bodies) while microgamonts (male gamonts) withnumerous nuclei (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e).Histology of mesenteric lymph node reveled lymhadenitis with infiltration of neutrophils and macropahges at places in one case while varying depletion of lymphoid cells and congestion in both the cases.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn the current study, we observed only 2.27% (16) Eimeria prevalence which was in line with the finding of Brar et al. (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) and Hussain et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) who reported only 4 and 2.9% prevalence respectively. In contrast to our finding, most of the researchers around the globe reported much higher prevalence ranging from 6.0%to 91.7% (Manya et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2008\u003c/span\u003e;Das et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e;Girma 2017;Nain et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2017\u003c/span\u003e;Morgoglione et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The reason for getting very low prevalence in our study may be the age of the calf from which samples were collected and prepatient period of parasite (Mundt et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In our study we collected 87.14% samples from less than three week of age group animals. We did not find any positive case in first week of age and only one case in second week. Another reason may be easy availability of effective drugs against this parasite in this area and increased awareness about hygienic practices. Li et al. (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) did meta-analysis of online available data of coccidial prevalence in China upto February 2020 and concluded reducing trend of prevalence year after year (from 57.9% before 2000 to 25.0% after 2015). Morgoglione et al.(2020) reported higher prevalence both in summer (82.4% and winter (82.3%)) season in buffalo calveswhile Das et al (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) reported highest prevalence of \u003cem\u003eE. bovis\u003c/em\u003e in pre monsoon (62.8%), followed by monsoon (58.6%), winter (55.4%) and summer season(50.8%).Higher prevalence observed in male calves in current study is in agreement with most of the previous researchers (Nain et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2017\u003c/span\u003e;Heidari et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).Though we found higher prevalence in older group of animals and male sex but statistically (Chi square test) it is non-significant. Direct life cycle of this parasite (without any intermediate host) and zoonotic implication explains the non-significant association of the cryptosporidium with above the risk variables.\u003c/p\u003e \u003cp\u003eSimilar to our study Mundt et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) also reported watery to haemorrhagic diarrhea but they observed lesions starting from terminal jejunum upto mid colon in experimentally infected calves. We observed highest severity of gross lesions in ileum and caecum. In contrast, Pyziel et al. (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) recorded gross lesions mainly in large intestine. Hussain et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) noted specific transverse zebra striping on the mucosal surface in all the case along with haemorrhage and necrosis. White raised nodules have been reported in small ruminants (Satish et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) but non apparent in bovine coccidiosis. Variable pathology of different species of Eimeria has been reported by various scientists (Das et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2015\u003c/span\u003e;Cruvinel et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eOur histological findings are in line with Hussain et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) who also documented mononuclear cellular infiltration in the lamina propria, desquamation of epithelial lining, and severe hemorrhages with mucosal scars and coccidial cells nested in the lamina propria in all infected animals. Mundt et al. (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) reportedmicroscopic lesions starting from initial mucosal hyperemia to acute typhlitis and necrotising colitis. They also observed infiltration of granulocytic series and proliferation of goblet cell. However, Sultana et al. (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) reported highest severity both in caecum and colon. They observed mild hyperemia along with granulocytic mucosal infiltration at 15 day post infection (dpi) and acute sever lesion (degeneration, shortened and necrosis of villi, haemorrhages in mucosa and sub-mucosa, widening of crypts of lieburkuhn and oedema of lamina propria) at 25dpi. Stockdale (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e1977\u003c/span\u003e) also reported similar findings along with fibrin deposits and diphtheritic membrane on mucosa at 18dpi followed by resolution and mucosal regeneration commencing from 22nd to 32nd dpi but they observed lesions mainly in large intestine.Kawahara (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) also observed similar lesion produced by gametogony both in small and large intestine. The lesions observed in our study such as severe congestion destruction of villus epithelium was mainly attributed to gametogony which can be seen in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e1\u003c/span\u003e by presence of different developmental stages of Eimeria deep into submucosa. As a result of gametogony there was massive destruction and stunting of villi. In an effort of regeneration, there was proliferation of crypt epithelium. Oedema of mucosa appeared due damage of villus epithelium as well as leaky blood vessels of lamina propria. The absorptive mature villus epithelium was replaced by immature crypt epithelium with secretory activity which causes diarrhea and when there is destruction of villi, it leads to haemorrhage and dysentery. The invasion of Eimeria to submucosalpayers patches of current study have not been reported by previous researches. Most of the scientists reported mucosal involvement only (Mundt et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2005\u003c/span\u003e;Sultana 2015;Hussain et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eBovine coccidiosis is an endemic with low prevalence but with severe onset of disease.Histology revealed its predilection of distal small intestine and proximal large intestine. The gross pathology revealed lesions varying from mild congestion to haemorrhagic enteritis, necrosis, ballooning and enlargement of mesenteric lymph node. Microscopically, lesions include congestion, mucosal oedema, mononuclear cell infiltration, destruction and stunting of villus epithelium. Parasitic gametogony responsible for villus destruction, cryptic dilatation, accumulation of necrotic debris and replacement of mature villus epithelium with immature secretory cryptic epithelium leading to malabsorption, diarrhea, dehydration and death. Coccidial prevalence was not found to be associated with age, sex and season.\u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgement:\u0026nbsp;\u003c/strong\u003eThe authors acknowledge the Science \u0026amp; Engineering Research Board (SERB), Department of Science \u0026amp; Technology, Government of India for providing funds through project entitled \u0026ldquo;Biomolecular detection of cryptosporidium and associated etiological agents from faecal samples of diarrheic bovine calves\u0026rdquo; project no. EMR/2017/000540/AS\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003eScience \u0026amp; Engineering Research Board (SERB), Department of Science \u0026amp; Technology, Government of India, Project no. EMR/2017/000540/AS\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompliance with ethical standards:\u003c/strong\u003e\u0026nbsp; The field study was approved by the Institutional ethical committee - IAEC/2019/188-221.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting Interests:\u0026nbsp;\u003c/strong\u003eThe authors of this research article declare that they have no conflict of interest\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eUndertaking\u003c/strong\u003e: The manuscript has not been published previously in full form or partially, and is not under consideration for publication elsewhere.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors contribution:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e1. Dr. Vikas Jaiswal: Sample collection, processing and manuscript writing\u003c/p\u003e\n\u003cp\u003e2. Dr.Apminder Pal Singh Brar: Designed the project, execution, monitoring and statistical analysis (project principal investigator)\u003c/p\u003e\n\u003cp\u003e3. Dr.Bhupinder Singh Sandhu: Photomicrography, micrometry and histopathological evaluation\u003c/p\u003e\n\u003cp\u003e4. Lachhman Das Singla: Parasitic identification and manuscript editing\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBrar A, Sood NK, Kaur P, Singla LD, Sandhu BS, Gupta K, Narang D, Singh CK, Chandra M (2017) Periurban outbreaks of bovine calf scours in Northern India caused by Cryptosporidium in association with other enteropathogens. Epidemiol Infect 145(13):2717\u0026ndash;2726. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1017/S0950268817001224\u003c/span\u003e\u003cspan address=\"10.1017/S0950268817001224\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCruvinel LB, Nicaretta JE, Bastos T, Couto L, Santos J, Zapa D, Cavalcante A, Cruz BC, BorgesD, Borges FA, Soares VE, Lopes W (2018) Eimeria species in dairy and beef cattle of different ages in Goi\u0026aacute;s state, Brazil. 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Vet World 14(9):2339\u0026ndash;2345. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.14202/vetworld.2021.2339-2345\u003c/span\u003e\u003cspan address=\"10.14202/vetworld.2021.2339-2345\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGirmaM (2017) Prevalence of Eimeria infection in calves in and around Sekota town, North Wollo, Ethiopia. Int j adv res 4(8):169\u0026ndash;175\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHeidari H, Sadeghi-Dehkordi Z, Moayedi R, Gharekhani J (2014) Occurrence and diversity of Eimeria species in cattle in Hamedan province. Iran Vet Med 59:271\u0026ndash;275\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHussain R, Mahmood F, Khan A, Mehmood K (2017) Prevalence and pathology of bovine coccidiosis in Faisalabad district, Pakistan. Thai J Vet Med 47(3):401\u0026ndash;406\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKawahara F (2016) Parasitological analyses of Eimeria infections in domestic animals and the development of molecular methods for species discrimination. Doctoral thesis Hokkaido University.\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.14943/doctoral.r6982\u003c/span\u003e\u003cspan address=\"10.14943/doctoral.r6982\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi DL, Gong QL, Ge GY, Wang Q, Sheng CY, Ma BY, Chen ZY, Yang Y, Li JM, Shi K, Leng X, Du R (2021) Prevalence and infection risk factors of bovine Eimeria in China: a systematic review and meta-analysis. Parasite 28:61. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1051/parasite/2021055\u003c/span\u003e\u003cspan address=\"10.1051/parasite/2021055\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eManya P, Sinha SRP, Sinha Sucheta, Verma SB, Sharma SK, Mandal KG (2008) Prevalence of bovine coccidiosis at Patna. Vet Parasitol 22(2):73\u0026ndash;76\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMorgoglione ME, Bosco A, Maurelli MP, Alves LC, Saralli G, Bruni G, Cringoli G, Rinaldi L (2020) A 10-Year Surveillance of Eimeria spp. in Cattle and Buffaloes in a Mediterranean Area. Front vet sci 7:410. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.3389/fvets.2020.00410\u003c/span\u003e\u003cspan address=\"10.3389/fvets.2020.00410\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMundt HC, Bangoura B, Rinke M, Rosenbruch M, Daugschies A (2005) Pathology and treatment of Eimeria zuerniicoccidiosis in calves: investigations in an infection model. Parasitol Int 54(4):223\u0026ndash;230. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.parint.2005.06.003\u003c/span\u003e\u003cspan address=\"10.1016/j.parint.2005.06.003\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNain N, Gupta SK, Sangwan AK, Gupta S (2017) Prevalence ofeimeria species in buffalo calves of Haryana Haryana Vet. 56:5\u0026ndash;81\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eOlabode VB, Gunya DY, Alsea UM, Choji TPP, Barde IJ (2020) Histopathological Lesions of Coccidiosis Natural Infestation in Chickens. Asian J Res Anim Veterinary Sci 5(2):41\u0026ndash;45\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePyziel AM, Demiaszkiewicz AW, Osińska B, Dolka I, Anusz K, Laskowski Z (2020) Usefulness of PCR-RFLP of 18S rRNA gene for rapid post-mortem diagnostics of highly pathogenic Eimeria spp. (Apicomplexa: Eimeriidae) of European bison, Bison bonasus L. with histopathological correlation. Int J Parasitol Parasites Wildl 12:13\u0026ndash;18. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1016/j.ijppaw.2020.03.008\u003c/span\u003e\u003cspan address=\"10.1016/j.ijppaw.2020.03.008\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRamirez L, Berto B, Filho W, Flausino W, Meireles G, Rodrigues J, Almeida C, Lopes C (2009) Eimeria bareillyi from the domestic water buffalo, Bubalusbubalis, in the State of Rio de Janeiro, Brazil. Revistabrasileira de medicinaveterinaria. 31. 261\u0026ndash;264\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRehman TU, Khan MN, Sajid MS, Abbas RZ, Arshad M, Iqbal Z (2011) Epidemiology of Eimeria and associated risk factors in cattle of district Toba Tek Singh, Pakistan. Parasitol Res 108(5):1171\u0026ndash;1177. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://dx.doi.org/10.1007/s00436-010-2159-5\u003c/span\u003e\u003cspan address=\"10.1007/s00436-010-2159-5\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSatish A, Nagarajan K, Balachandran C, Soundararajan C, Legadevi R (2019) Gross and Histopathology of Coccidiosis in Small Ruminants in Tamil Nadu. Int J Livest Res 9(2):225\u0026ndash;235. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.5455/ijlr.20180603054413\u003c/span\u003e\u003cspan address=\"10.5455/ijlr.20180603054413\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStockdale PH( (1977) The pathogenesis of the lesions produced by Eimeria zuernii in calves. Can J Comp medicine: Revue canadienne de medecinecomparee 41(3):338\u0026ndash;344\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSultana R, Maqbool A, Ahmad MZ, Ahmed MD, Ch IZ (2015) Histopathological Studies of Eimeria bovis Infection in Calves. J Veterinary Sci Technol 06(04). doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4172/2157-7579.1000235\u003c/span\u003e\u003cspan address=\"10.4172/2157-7579.1000235\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTamrat H, Mekonnen N, Ferede Y, Cassini R, Belayneh N (2020) Epidemiological study on calf diarrhea and coccidiosis in dairy farms in Bahir Dar, North West Ethiopia. Ir veterinary J 73:14. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s13620-020-00168-w\u003c/span\u003e\u003cspan address=\"10.1186/s13620-020-00168-w\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWalker RA, Ferguson DJ, Miller CM, Smith NC (2013) Sex and Eimeria: a molecular perspective. Parasitology 140(14):1701\u0026ndash;1717. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1017/S0031182013000838\u003c/span\u003e\u003cspan address=\"10.1017/S0031182013000838\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePrevalence of Eimeria in different season, sex and age group of bovine calves.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eVariable\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo of samples\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eNo of positive samples\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePrevalence\u003c/p\u003e \u003cp\u003e(%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSeason\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWinter\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e178\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.80\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSummer\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e125\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMonsoon\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e210\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.38\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePost monsoon\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e187\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003e\u003cb\u003eAge group\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup I (0\u0026ndash;7 day)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e221\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup II (8\u0026ndash;14 day)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e312\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.32\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup III (15\u0026ndash;21 day)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.19\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup IV (22\u0026ndash;30 day)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.31\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup V (31\u0026ndash;60 day)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.00\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSex\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e368\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.72\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e332\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.80\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eTotal\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e700\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e16\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e2.29\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eSignificant relationship of variables (season, age and aex) with Eimeria spp. infection in faecal samples of calves.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eEtiological agent\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eSeason\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eAge\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eSex\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eChi-Square\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ep-Value\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eChi-Square\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ep-Value\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eChi-Square\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003ep-Value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eEimeria\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8.533\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.036\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4.668\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.458\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.528\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.467\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.01 considered significant.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"journal-of-parasitic-diseases","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jopd","sideBox":"Learn more about [Journal of Parasitic Diseases](https://www.springer.com/journal/12639)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/jopd/default.aspx","title":"Journal of Parasitic Diseases","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Eimeria, Prevalence, histopathology","lastPublishedDoi":"10.21203/rs.3.rs-2456431/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2456431/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eBovine coccidiois, caused by Eimeria spp. is widely prevalent around the globe and responsible for huge economic losses by causing morbidity and mortality among young calves.The present study was designed to evaluate the prevalence as well as to evaluate histopathological alteration associated with it. The faecal samples were collected from 700 bovine calves uptotwo month of age from August 2019 to July 2021 and screened for Eimeria oocycts. The intestinal tissue samples of 37 calves were also collected which died during the study period after showing symptoms of diarrhea and examined for histological lesions.The faecal prevalence of Eimeria observed in our study was 2.29% (16/700) while in tissue samples only two out of 37 were foundpositive for Eimeria infection. Tissue sections reveled various stages of Eimeria gametogony, variable congestion, haemorrhage, and necrosis along with cryptic dilatation and mononuclear cell infiltration.Coccidia was not found to be associated with season, age and sex of calf. Bovine coccidiosiswas found to be endemic with low prevalence but severe onset characterized by moderate to severe congestion and inflammatory reaction mainly in the ileum and caecum.\u003c/p\u003e","manuscriptTitle":"Faecal prevalence and histopathological evaluation of coccidiosis in bovine calves","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-01-17 21:17:00","doi":"10.21203/rs.3.rs-2456431/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"reviewerAgreed","content":"","date":"2023-01-16T11:09:01+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2023-01-16T05:53:09+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2023-01-09T06:08:12+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Parasitic Diseases","date":"2023-01-08T12:45:18+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-parasitic-diseases","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"jopd","sideBox":"Learn more about [Journal of Parasitic Diseases](https://www.springer.com/journal/12639)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/jopd/default.aspx","title":"Journal of Parasitic Diseases","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"bae30060-8420-45a1-a5d9-309f01337c1c","owner":[],"postedDate":"January 17th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2023-10-16T21:20:53+00:00","versionOfRecord":{"articleIdentity":"rs-2456431","link":"https://doi.org/10.1007/s12639-023-01590-x","journal":{"identity":"journal-of-parasitic-diseases","isVorOnly":false,"title":"Journal of Parasitic Diseases"},"publishedOn":"2023-05-24 20:58:54","publishedOnDateReadable":"May 24th, 2023"},"versionCreatedAt":"2023-01-17 21:17:00","video":"","vorDoi":"10.1007/s12639-023-01590-x","vorDoiUrl":"https://doi.org/10.1007/s12639-023-01590-x","workflowStages":[]},"version":"v1","identity":"rs-2456431","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2456431","identity":"rs-2456431","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}