Phylogenetic study of Dipylidium caninum Isolated from cats (Felis domestica) and dogs (Canis familiaris) intestines.

Phylogenetic study of Dipylidium caninum Isolated from cats (Felis domestica) and dogs (Canis familiaris) intestines. | 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 Short Report Phylogenetic study of Dipylidium caninum Isolated from cats (Felis domestica) and dogs (Canis familiaris) intestines. Musafer Al-Ardi This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4535872/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Knowing the parasites' genotypes and establishing their phylogenetic tree help to define their route of evolution and determine their origin. On the other hand, understanding the variation in genome sequences aids in the development of techniques for identifying diseases and selecting the best and most efficient treatments. This study used worms isolated from cats and dogs intestines to genotyping and draw phylogenetic tree of Dipylidium caninum . This study approach involved isolating worms directly from the intestines of dogs and cats, and confirming infection through molecular diagnosis using primers designed by the author. Genotyping using primers for the large subunit rRNA gene revealed sequences of DNA products that were deposited in the GenBank. Finally, these sequences were aligned with previous sequences deposited in the GenBank, and gene sequences in cats were compared with their counterparts in dogs. Infection significantly were higher among dogs than cats and among males than females. Three sequences (OQ987885 to OQ987887) isolated from dogs and another (OQ987888 to OQ987890) isolated from cats have been deposited in the Gen Bank. Phylogenetic tree comparing these sequences and showing the genetic distance between them. Sequences deposited from this study recorded high alignment with (MG587892.1) and (OK523385.1) sequences dating back to D. caninum . The host species influences the species and ability of the parasite that will infect it, and possibly the genotypes as well. Canine fleas can infect dogs and cats, while feline fleas can infect cat only. so the parasite must adapt to continue living, and one of the ways to adapt is heterogeneity of genotypes. Dipylidium caninum Genotyping Felis catus rRNA tapeworm Figures Figure 1 Figure 2 Figure 3 Introduction One of the parasitic worms that has deeply affected animals since ancient times is Dipylidium caninum . Ancient Babylonian humans were the first reported infections (Lefkaditis and Koukeri, 2006 ). As their definitive host, it infects members of the Felidae as cats and Canidae families, including foxes, and dogs (Fourie et al. 2013 ). These worms have a two-stage life cycle, the first stage occurring as a larva in the abdominal of fleas ( Ctenocephalides canis and C. felis ) or lice (Yadav et al. 2019 ). The second stage is the adult, which parasitizes Carnivores (Gutema et al. 2020 ). A mild tapeworm infection is asymptomatic. Anorexia, epigastric pain, diarrhoea, weight loss, pruritus, anal pruritus, urticarial, and constipation are symptoms of moderate infections (Ramos et al. 2020 ). Infected cats can exhibit severe symptoms such as intestinal obstruction, convulsions, and epileptic seizures (Benitez‑Bolivar et al. 2022). D. caninum with the two genital pore loaded with egg pockets gravid proglottids can be easily identified in the sample using the naked eye or a hand lens (Chong et al. 2020 ). However, wet smear microscopy confirms infection more frequently (García-Agudo et al 2014 ). As a result, molecular diagnosis was consider less interesting for D. caninum identification. Despite their accuracy, molecular diagnostic methods are only used to diagnosing infection in the definitive host (Labuschagne et al 2018 ). There are many studies looking at genotypes of the genus Dipylidium (Liu et al. 2021 ). In addition, they look at the mitochondrial genome and not the complete genome of the organism (Beugnet et al. 2014 ) Historically, many species were included in the genus Dipylidium after studying its morphological structure (Labuschagne et al. 2018 ), but the molecular studies were limited to one species, D. caninum , and considered it monotype (Izabela et al. 2020 ). In this same biography, genotyping may have an opinion in proving the existence of many genotypes belonging to this species (Labuschagne et al. 2018 ). Instead of using faeces and fleas, the current study focused on genotyping D. caninum isolated from the guts of dogs and cats. On the other hand, answering the question of whether the genotypes of the worm that infects cats and dogs are identical or distinct. Materials and method From January 2022 to June 2023, under the guidance of a qualified veterinarian, sixty-nine road-killed animals (37 cats and 32 dogs) from various locations in the Al-Qadisiyah Governorate (31.17 N, 44.24 E) were collected. Age and gender for animals were created. Ethics Approval The Ethics Committee at Ministry of Agriculture - Veterinary Department No. 99 at 15/12/2022, accepted ethic statements for collection samples from animals. Parasites collection The intestine was cut longitudinally with scissors after the autopsy by the vet. Some worms were picked up, and washed with distilled water twice time. The naked eye was used to verify the worms. White gravid proglottids with a cucumber-like shape were seen through a hand lens. The parasite genus was identified using a wet smear and a light microscope. All worms were stored at -20°C in distilled water (Izabela et al. 2020 ) . Primers design: Bioneer company, Korea designed all primers that sequenced as following: Primer Sequnces Size Ref. mtDNA F: 5′- CTATTGATTGCGTTTATTGTTTTGTGT-3′ 290 Pb Zhu et al., 2019 R: 5′-GAAAAGAAATCAAATACAGTTAAACGGT-3′ DFC28S-1R rRNA F: 5′-CAC ATT CAA CGC CCGACT CCT GTA G-3′ 653 Pb Benitez‑Bolivar et al , 2022 R: 5'GAAAAGAAAA- AATACAGTTACG 3' Molecular analysis Bioneer Korean company developed all Primer. The infestation and the parasite genus were verified by the mtDNA gene amplification in accordance with Zho, et al ., (2019). The large subunit rRNA gene was amplified in the positive samples using its primers, and the outcomes were transferred to the Bioneer Company for sequencing using the sanger dideoxy method. Phylogenetic tree analysis With specific accession numbers, the genotypes were deposited in a gene bank. After then, these genotypes were aligned to those that had been deposited in the gene bank. Then, using MEGA version 6 to draw phylogenetic tree using the likelihood maximum tree test (Kumar et al. 2016 ). Statistical analysis The mean, standard deviation, p values, and confidence intervals were extracted using the statistical software spss24. Results The infection rates for cats and dogs were (8.1%) and (31.25%), respectively, as shown in Tables 1 and 2 . According to the two tables, it is evident that older cats had greater infection rates than younger (20%), however in dogs, 2–3 year olds had contracted the disease (40%). With the naked eye and a hand lens, intestinal debris was discovered to contain a white, one-centimetre-long structure that like a rice grain. An examination under a microscope made it clear that there were D. caninum infestations. The gravid proglottids had lengths of 12 to 15 mm and a thickness of 2 to 4 mm. When the gravid segments were crushed, it was discovered that they contained 8–12 eggs that were maintained in pockets and covered by a thin membrane. The results of a molecular analysis demonstrated that all D. caninum infections were present (Fig. 1 ). For sequencing, a large number of DNA amplification products were provided. Isolated sequences from dogs and cats were each given the sequence numbers (OQ987885 to OQ987887) in dogs and (OQ987888 to OQ987890) in cats, which deposited in the Gen Bank, respectively. The deposited sequences from this study recorded high alignment with (MG587892.1) and (OK523385.1) (Fig. 2 , Table 3 ). On the other hand, genotyping showed that the genotypes that affect cats are not identical to those that affect dogs despite the presence of a close relationship between them, which indicates the presence of two distinct genotypes or even two diverse species (Fig. 3 , Table 4 ). Table 1 Infection rates and cat traits in the experiment Parameters No. of case infected Odd ratio 95% IC: p.value Gender Male Female Age > 1year 1–2 years > 2 years 15(40.5%) ± 1.17 22(59.5%) ± 1.12 14(37.8) ± 1.42 10(27%) ± 1.33 13(35.2%) ± 2.06 1(6%) 2(9%) 0(0%) 2(20%) 1(7%) 1.36 2.6154 0.113– 16.42 0.205–32.09 0.083 0.048 Total 37(100%) 3(8.1%) Table 2 Infection rates and dog traits in the experiment Parameters No. of case infected Odd ratio 95% IC: p.value Gender Male Female Age 1–2 year 2–3 years > 3 years 15(40.5%) ± 1.17 17(59.5%) ± 1.12 14(37.8) ± 1.42 10(27%) ± 1.33 8(35.2%) ± 2.06 5(33.3%) 5(29.4%) 3(21.4%) 4(40%) 3(37.5%) 0.88 0.93 0.213– 3.65 0.16–546 0.083 0.094 Total 32(100%) 10(31.25%) Table 3 The genotypes of D. caninum in dogs and cats using partial sequence of rRNA Gene according to phylogenetic tree analysis and NCBI- BLAST alignment tool. Dogs isolates Isolate 1 Isolate 2 Isolate 3 Max Score Per. Ident. Max Score Per. Ident. Max Score Per. Ident. MH045481.1 242 97.87 231 95.77 245 99.28 OL460638.1 153 85.31 359 88.33 159 86.52 Cats Isolates Isolate 1 Isolate 2 Isolate 3 Per. Ident. Max Score Per. Ident. Per. Ident. Max Score Per. Ident. MH045481.1 250 98.58 244 97.87 244 97.87 OL460638.1 161 88.32 156 87.68 156 87.59 genotypes among other genotypes. Table 4 The percentage of genetic distances among sequences of cats and dogs. Isolate No. Dog1 Dog2 Dog3 Cat1 Cat2 Cat3 Dog1 Dog2 0.00 Dog3 0.00 0.00 Cat1 3.00 3.00 6.25 Cat2 3.25 3.25 3.25 3.00 Cat3 3.00 3.00 6.25 0.00 3.00 Discussion External environmental factors and host preferences influence D. caninum infection (Liuet al. 2021 ). Consequently, even though males are more likely to develop an infection than females are, this difference is an insignificant (Fourie et al. 2013 ). Similar to how infection varies with age, it actually increases slightly in younger animals (Trasviña-Muñoz et al. 2020 ) due to older enhanced immune activity (East et al. 2103). In spite some studies, the prevalence is rising among the elderly (Cantó et al. 2011 ). The final host's lifestyle has an impact on the infection rate. Lack of care for stray and shelter animals causes infection rates to grow among them (Zottler et al. 2019 ) especially when intermediate hosts (Abdullah et al. 2019 ). Because of cats' self-care behaviour, dogs may consume fewer fleas (Beugnet et al. 2014 ), but cats experience fewer flea infestations than dogs (García-Agudo et al. 2014 ). The hand lens or a microscope can be utilized to set eye on the worm, but a molecular diagnostic using specialised primers for specific genes yields more accurate and trustworthy results. While prior research were intended to identify the parasite in isolated flea samples (Low et al. 2017 ), Zhu et al. ( 2019 ) confirmed the feasibility of recovering DNA from parasite eggs isolated from dog feces. Beugnet et al. ( 2014 ) revealed the presence of two unique sequences from 28S rDNA region when amplified a region DNA extracted from canine and feline fleas by PCR technique. He explained this by the existence of two distinct genotypes, each of which infects t flea species ( C. canis or C. felis ) (Beugnet et al. 2014 ). In line with this, Labuschagne et al. ( 2018 ) created genotyping assays based on hydrolysis probes and restriction fragment length polymorphism (RFLP) analyses to identify and classify D. caninum isolated from fleas. In order to find D. caninum DNA in cat’s faeces, he used a mitochondrial genome. He did this to demonstrate that D. caninum infections are highly particular to the host. He confirmed that the feline genotype was more common in cats than in dogs, with the canine genotype being more common in dogs (Labuschagne et al. 2018 ). In a similar manner, none of the gravid proglottids that have been isolated from feces samples exhibits any evidence of hybridization, despite the fact that various adult D. caninum strains are capable of developing in the host intestine (Labuschagne et al. 2019). In Malaysia, it was established that the genotype of D. caninum isolated from cats perfectly matches the DNA obtained from fleas on cats with tapeworm. They are two discrete clades that belong to two phenotypes that infect cats and dogs and are found in different parts of the world with differing morphological characteristics. They did not adhere to the regional distribution pattern or a particular geographic location (Low et al. 2017 ). A brand-new type of tapeworm known as D. carracidoi was discovered in Spain. Its similarity to D. caninum as determined by phenotypic research may indicate that it is a different genotype because of the latter species (Millán and Casanova 2009 ). In conclusion, the gravid proglottids of worms isolated from definitive hosts can be used for molecular diagnosis and genotyping. D. caninum exists in two different genotypes, one infecting cats and the other in dogs. The use of the RFLP approach and restriction enzymes in the three parasite positions (intestine, feces, and fleas) will probably give results that are more accurate. Declarations Author Contribution All paper parts were done y Musafer AlArdi Acknowledgement Al-Hamza Veterinary Hospital staff, especially Dr. Amin Abdulameer, I really appreciate your help. Conflict of interest. The author published this article and has no direct interest in these studies other than publishing the scientific findings. References Abdullah S, Helps C, Tasker S, Newbury H, Wall R (2019) Pathogens in fleas collected from cats and dogs: Distribution and prevalence in the UK. 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Case Rep Pediatr 1–3. 10.1155/2020/4716124 East ML, Kurze C, Wilhelm K, Benhaiem S, Hofer H (2013) Factors influencing Dipylidium sp. infection in a free-ranging social carnivore, the spotted hyaena (Crocuta crocuta). Int J Parasitol Parasites Wildl 2(1):257–265. 10.1016/j.ijppaw.2013.09.003 Fourie JJ, Crafford D, Horak IG, Stanneck D (2013) Prophylactic treatment of flea-infested dogs with an imidacloprid / flumethrin collar (seresto®, bayer) to preempt infection with dipylidium caninum. Parasitol Res 112(1):33–46. 10.1007/s00436-013-3279-5 García-Agudo L, García-Martos P, Rodríguez-Iglesias M (2014) Dipylidium Caninum Infection in an Infant: A Rare Case Report and Literature Review. Asian Pac J Trop Biomed 4:565–567. 10.12980/APJTB.4.2014APJTB-2014-0034 Gutema FD, Yohannes GW, Abdi RD et al (2020) Dipylidium caninum Infection in Dogs and Humans in Bishoftu Town, Ethiopia. Diseases . 2020;9(1):1. 10.3390/diseases9010001 Izabela W, Sławomir AP, Kornás KR (2020) The Prevalence of Endoparasites of Free Ranging Cats. Animals 10(1):748–761. 10.3390/ani10040748 Kumar S, Stecher G, Tamura K (2016) MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol 33(7):1870–1874. 10.1093/MOLBEV/MSW054 Labuschagne M, Beugnet F, Rehbein S, Guillot J, Fourie J, Crafford D (2018) Analysis of Dipylidium caninum tapeworms from dogs and cats, or their respective fleas: Part 1. Molecular characterization of Dipylidium caninum: Genetic analysis supporting two distinct species adapted to dogs and cats. Parasite 25(30):1–17. 10.1051/parasite/2018028 Lefkaditis AM, Koukeri SE (2006) Prevalence of Dipylidium caninum, in domestic cats from the city of Thessaloniki, Greece. Rev Sci Parasitol 7(1/2):100–103. https://www.cabdirect.org/cabdirect/abstract/20073009184 Liu GH, Korhonen PK, Young ND et al (2021) Dipylidium caninum draft genome - a new resource for comparative genomic and genetic explorations of flatworms. Genomics 113(3):1272–1280. 10.1016/j.ygeno.2021.02.019 Low VL, Prakash BK, Tan TK et al (2017) Pathogens in ectoparasites from free-ranging animals: Infection with Rickettsia asembonensis in ticks, and a potentially new species of Dipylidium in fleas and lice. Vet Parasitol 245(1):102–105. 10.1016/j.vetpar.2017.08.015 Millán J, Casanova JC (2009) High prevalence of helminth parasites in feral cats in Majorca Island (Spain). Parasitol Res 106(1):183–188. 10.1007/s00436-009-1647-y Ramos N, de Silva V, Barreto MLE, Barros MS, Mendes-De-almeida LA F (2020) Endoparasites of household and shelter cats in the city of Rio de Janeiro, Brazil. Rev Bras Parasitol Vet 29(1):1–15. 10.1590/S1984-29612019110 Trasviña-Muñoz E, López-Valencia G, Monge-Navarro FJ et al (2020) Detection of intestinal parasites in stray dogs from a farming and cattle region of northwestern mexico. Pathogens 9(7):516–524. 10.3390/pathogens9070516 Zhu GQ, Li L, Ohiolei JA et al (2019) A multiplex PCR assay for the simultaneous detection of Taenia hydatigena, T. multiceps, T. pisiformis , and Dipylidium caninum infections. BMC Infect Dis 19(1):854–867. 10.1186/s12879-019-4512-3 Zottler EM, Bieri M, Basso W, Schnyder M (2019) Intestinal parasites and lungworms in stray, shelter and privately owned cats of Switzerland. Parasitol Int 69(1):75–81. 10.1016/j.parint.2018.12.005 Yadav S, Kumar Patel P, Deepak D, Bhatt S, Kumari Patel S, Dixit S (2019) Medical Management of Dipylidium Caninum Infected Female Cat: A Case Report. Int J Chem Stud 7:550–552. https://www.chemijournal.com/archives/2019/vol7issue1/PartJ/6-6-339-723.pdf Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted 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-4535872","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Short Report","associatedPublications":[],"authors":[{"id":312990644,"identity":"10ce1496-b89a-4165-b21b-97ffb861d63a","order_by":0,"name":"Musafer Al-Ardi","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA10lEQVRIiWNgGAWjYNACGwkG++PNBxgYG4jWkibBwHDmWAJJWoD4Ro4BcVp0Z6Q//vAjwSKfsefMN4mfO2zkGNgPH92AT4vZjRwzyZ4ECctm9t5tkr1n0owZeNLSbhDQwsbA+0PCgI3n7DYJ3rbDiQ0SPGYEtKQ//vgnQcKARyLnmeRf4rQkGEjzALVISOSwSRNny5k3ZtIyQC0GPMeMrWXb0ozZCPrlONBhbxLqDAzYmx/efNtmI8fPfvgYXi3IgEUCRLIRqxwEmD+QonoUjIJRMApGDgAAi11KVwe+5ekAAAAASUVORK5CYII=","orcid":"","institution":"Ministry of Higher Education and Scientific Research","correspondingAuthor":true,"prefix":"","firstName":"Musafer","middleName":"","lastName":"Al-Ardi","suffix":""}],"badges":[],"createdAt":"2024-06-05 18:48:56","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4535872/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4535872/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":59045646,"identity":"37f150f2-0674-4897-91a2-ad21b06bfad8","added_by":"auto","created_at":"2024-06-25 18:19:24","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":12726,"visible":true,"origin":"","legend":"\u003cp\u003emt DNA gene amplification products after electrophoresis using agarose gel.\u003c/p\u003e","description":"","filename":"Picture1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4535872/v1/4e49980bf774a67804b17ea9.jpg"},{"id":59045855,"identity":"9da8ae35-6de2-4eb0-ae8d-aade6bc410ad","added_by":"auto","created_at":"2024-06-25 18:27:24","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":23383,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree analysis by Maximum Likelihood test \u0026nbsp;illustrates position of study genotypes among other genotypes.\u003c/p\u003e","description":"","filename":"Picture2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4535872/v1/01658e1fd7b251b08b90906a.jpg"},{"id":59045648,"identity":"ba96d702-926c-40bc-aa32-db4eb0e6530b","added_by":"auto","created_at":"2024-06-25 18:19:24","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":11180,"visible":true,"origin":"","legend":"\u003cp\u003ePhylogenetic tree analysis by Maximum Likelihood test \u0026nbsp;illustrates divergent cats and dogs \u0026nbsp;\u003cem\u003eD. caninum\u003c/em\u003egenotypes.\u003c/p\u003e","description":"","filename":"Picture3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4535872/v1/24cdfb61c4906ecae1f5f4c8.jpg"},{"id":73912813,"identity":"dffa145d-00fa-4569-8880-56256eaec487","added_by":"auto","created_at":"2025-01-16 00:01:22","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":611453,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4535872/v1/25d0c09c-f075-4d55-b334-ed70d2ecd6e4.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Phylogenetic study of Dipylidium caninum Isolated from cats (Felis domestica) and dogs (Canis familiaris) intestines.","fulltext":[{"header":"Introduction","content":"\u003cp\u003eOne of the parasitic worms that has deeply affected animals since ancient times is \u003cem\u003eDipylidium caninum\u003c/em\u003e. Ancient Babylonian humans were the first reported infections (Lefkaditis and Koukeri, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). As their definitive host, it infects members of the Felidae as cats and Canidae families, including foxes, and dogs (Fourie et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). These worms have a two-stage life cycle, the first stage occurring as a larva in the abdominal of fleas (\u003cem\u003eCtenocephalides canis\u003c/em\u003e and \u003cem\u003eC. felis\u003c/em\u003e) or lice (Yadav et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The second stage is the adult, which parasitizes Carnivores (Gutema et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eA mild tapeworm infection is asymptomatic. Anorexia, epigastric pain, diarrhoea, weight loss, pruritus, anal pruritus, urticarial, and constipation are symptoms of moderate infections (Ramos et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Infected cats can exhibit severe symptoms such as intestinal obstruction, convulsions, and epileptic seizures (Benitez‑Bolivar et al. 2022).\u003c/p\u003e \u003cp\u003e \u003cem\u003eD. caninum\u003c/em\u003e with the two genital pore loaded with egg pockets gravid proglottids can be easily identified in the sample using the naked eye or a hand lens (Chong et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). However, wet smear microscopy confirms infection more frequently (Garc\u0026iacute;a-Agudo et al \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). As a result, molecular diagnosis was consider less interesting for \u003cem\u003eD. caninum\u003c/em\u003e identification. Despite their accuracy, molecular diagnostic methods are only used to diagnosing infection in the definitive host (Labuschagne et al \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThere are many studies looking at genotypes of the genus \u003cem\u003eDipylidium\u003c/em\u003e (Liu et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In addition, they look at the mitochondrial genome and not the complete genome of the organism (Beugnet et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) Historically, many species were included in the genus \u003cem\u003eDipylidium\u003c/em\u003e after studying its morphological structure (Labuschagne et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), but the molecular studies were limited to one species, \u003cem\u003eD. caninum\u003c/em\u003e, and considered it monotype (Izabela et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). In this same biography, genotyping may have an opinion in proving the existence of many genotypes belonging to this species (Labuschagne et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eInstead of using faeces and fleas, the current study focused on genotyping \u003cem\u003eD. caninum\u003c/em\u003e isolated from the guts of dogs and cats. On the other hand, answering the question of whether the genotypes of the worm that infects cats and dogs are identical or distinct.\u003c/p\u003e"},{"header":"Materials and method","content":"\u003cp\u003eFrom January 2022 to June 2023, under the guidance of a qualified veterinarian, sixty-nine road-killed animals (37 cats and 32 dogs) from various locations in the Al-Qadisiyah Governorate (31.17 N, 44.24 E) were collected. Age and gender for animals were created.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eEthics Approval\u003c/h2\u003e \u003cp\u003eThe Ethics Committee at Ministry of Agriculture - Veterinary Department No. 99 at 15/12/2022, accepted ethic statements for collection samples from animals.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eParasites collection\u003c/h2\u003e \u003cp\u003eThe intestine was cut longitudinally with scissors after the autopsy by the vet. Some worms were picked up, and washed with distilled water twice time. The naked eye was used to verify the worms. White gravid proglottids with a cucumber-like shape were seen through a hand lens. The parasite genus was identified using a wet smear and a light microscope. All worms were stored at -20\u0026deg;C in distilled water (Izabela et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) .\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003ePrimers design:\u003c/h2\u003e \u003cp\u003eBioneer company, Korea designed all primers that sequenced as following:\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Taba\" border=\"1\"\u003e \u003ccolgroup cols=\"4\"\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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePrimer\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSequnces\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSize\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eRef.\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003emtDNA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eF: 5\u0026prime;- CTATTGATTGCGTTTATTGTTTTGTGT-3\u0026prime;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e290 Pb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eZhu et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eR: 5\u0026prime;-GAAAAGAAATCAAATACAGTTAAACGGT-3\u0026prime;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eDFC28S-1R rRNA\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eF: 5\u0026prime;-CAC ATT CAA CGC CCGACT CCT GTA G-3\u0026prime;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e653 Pb\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eBenitez‑Bolivar \u003cem\u003eet al\u003c/em\u003e, 2022\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eR: 5'GAAAAGAAAA- AATACAGTTACG 3'\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\u003eMolecular analysis\u003c/p\u003e \u003cp\u003eBioneer Korean company developed all Primer. The infestation and the parasite genus were verified by the mtDNA gene amplification in accordance with Zho, \u003cem\u003eet al\u003c/em\u003e., (2019). The large subunit rRNA gene was amplified in the positive samples using its primers, and the outcomes were transferred to the Bioneer Company for sequencing using the sanger dideoxy method.\u003c/p\u003e \u003cp\u003ePhylogenetic tree analysis\u003c/p\u003e \u003cp\u003eWith specific accession numbers, the genotypes were deposited in a gene bank. After then, these genotypes were aligned to those that had been deposited in the gene bank. Then, using MEGA version 6 to draw phylogenetic tree using the likelihood maximum tree test (Kumar et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eThe mean, standard deviation, p values, and confidence intervals were extracted using the statistical software spss24.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eThe infection rates for cats and dogs were (8.1%) and (31.25%), respectively, as shown in Tables\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. According to the two tables, it is evident that older cats had greater infection rates than younger (20%), however in dogs, 2\u0026ndash;3 year olds had contracted the disease (40%).\u003c/p\u003e \u003cp\u003eWith the naked eye and a hand lens, intestinal debris was discovered to contain a white, one-centimetre-long structure that like a rice grain. An examination under a microscope made it clear that there were \u003cem\u003eD. caninum\u003c/em\u003e infestations. The gravid proglottids had lengths of 12 to 15 mm and a thickness of 2 to 4 mm. When the gravid segments were crushed, it was discovered that they contained 8\u0026ndash;12 eggs that were maintained in pockets and covered by a thin membrane.\u003c/p\u003e \u003cp\u003eThe results of a molecular analysis demonstrated that all \u003cem\u003eD. caninum\u003c/em\u003e infections were present (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). For sequencing, a large number of DNA amplification products were provided. Isolated sequences from dogs and cats were each given the sequence numbers (OQ987885 to OQ987887) in dogs and (OQ987888 to OQ987890) in cats, which deposited in the Gen Bank, respectively.\u003c/p\u003e \u003cp\u003eThe deposited sequences from this study recorded high alignment with (MG587892.1) and (OK523385.1) (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, Table \u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). On the other hand, genotyping showed that the genotypes that affect cats are not identical to those that affect dogs despite the presence of a close relationship between them, which indicates the presence of two distinct genotypes or even two diverse species (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, Table \u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \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\u003eInfection rates and cat traits in the experiment\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameters\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo. of case\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003einfected\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eOdd ratio\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e95% IC:\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\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\u003eGender\u003c/p\u003e \u003cp\u003eMale\u003c/p\u003e \u003cp\u003eFemale\u003c/p\u003e \u003cp\u003eAge\u003c/p\u003e \u003cp\u003e\u0026gt;\u0026thinsp;1year\u003c/p\u003e \u003cp\u003e1\u0026ndash;2 years\u003c/p\u003e \u003cp\u003e\u0026gt;\u0026thinsp;2 years\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15(40.5%)\u0026thinsp;\u0026plusmn;\u0026thinsp;1.17\u003c/p\u003e \u003cp\u003e22(59.5%)\u0026thinsp;\u0026plusmn;\u0026thinsp;1.12\u003c/p\u003e \u003cp\u003e14(37.8)\u0026thinsp;\u0026plusmn;\u0026thinsp;1.42\u003c/p\u003e \u003cp\u003e10(27%)\u0026thinsp;\u0026plusmn;\u0026thinsp;1.33\u003c/p\u003e \u003cp\u003e13(35.2%)\u0026thinsp;\u0026plusmn;\u0026thinsp;2.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1(6%)\u003c/p\u003e \u003cp\u003e2(9%)\u003c/p\u003e \u003cp\u003e0(0%)\u003c/p\u003e \u003cp\u003e2(20%)\u003c/p\u003e \u003cp\u003e1(7%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.36\u003c/p\u003e \u003cp\u003e2.6154\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.113\u0026ndash; 16.42\u003c/p\u003e \u003cp\u003e0.205\u0026ndash;32.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.083\u003c/p\u003e \u003cp\u003e0.048\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e37(100%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3(8.1%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c6\" namest=\"c4\"\u003e\u0026nbsp;\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\u003eInfection rates and dog traits in the experiment\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\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 \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameters\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNo. of case\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003einfected\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eOdd ratio\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e95% IC:\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\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\u003eGender\u003c/p\u003e \u003cp\u003eMale\u003c/p\u003e \u003cp\u003eFemale\u003c/p\u003e \u003cp\u003eAge\u003c/p\u003e \u003cp\u003e1\u0026ndash;2 year\u003c/p\u003e \u003cp\u003e2\u0026ndash;3 years\u003c/p\u003e \u003cp\u003e\u0026gt;\u0026thinsp;3 years\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15(40.5%)\u0026thinsp;\u0026plusmn;\u0026thinsp;1.17\u003c/p\u003e \u003cp\u003e17(59.5%)\u0026thinsp;\u0026plusmn;\u0026thinsp;1.12\u003c/p\u003e \u003cp\u003e14(37.8)\u0026thinsp;\u0026plusmn;\u0026thinsp;1.42\u003c/p\u003e \u003cp\u003e10(27%)\u0026thinsp;\u0026plusmn;\u0026thinsp;1.33\u003c/p\u003e \u003cp\u003e8(35.2%)\u0026thinsp;\u0026plusmn;\u0026thinsp;2.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5(33.3%)\u003c/p\u003e \u003cp\u003e5(29.4%)\u003c/p\u003e \u003cp\u003e3(21.4%)\u003c/p\u003e \u003cp\u003e4(40%)\u003c/p\u003e \u003cp\u003e3(37.5%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.88\u003c/p\u003e \u003cp\u003e0.93\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.213\u0026ndash; 3.65\u003c/p\u003e \u003cp\u003e0.16\u0026ndash;546\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.083\u003c/p\u003e \u003cp\u003e0.094\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTotal\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e32(100%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10(31.25%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"3\" nameend=\"c6\" namest=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe genotypes of \u003cem\u003eD. caninum\u003c/em\u003e in dogs and cats using partial sequence of rRNA Gene according to phylogenetic tree analysis and NCBI- BLAST alignment tool.\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\u003eDogs isolates\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eIsolate 1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eIsolate 2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eIsolate 3\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMax Score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePer. Ident.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMax Score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePer. Ident.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMax Score\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003ePer. Ident.\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMH045481.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e242\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e97.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e231\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e95.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e245\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e99.28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOL460638.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e153\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e85.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e359\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e88.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e159\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e86.52\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCats Isolates\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eIsolate 1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003eIsolate 2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c7\" namest=\"c6\"\u003e \u003cp\u003eIsolate 3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePer. Ident.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMax Score\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePer. Ident.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003ePer. Ident.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMax Score\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003ePer. Ident.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMH045481.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e250\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e98.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e244\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e97.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e244\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e97.87\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOL460638.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e161\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e88.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e156\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e87.68\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e156\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e87.59\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 \u003c/p\u003e \u003cp\u003egenotypes among other genotypes.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe percentage of genetic distances among sequences of cats and dogs.\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\"\u003e \u003cp\u003eIsolate No.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDog1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDog2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDog3\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eCat1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCat2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eCat3\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDog1\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 \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDog2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.00\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 \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDog3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCat1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCat2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCat3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eExternal environmental factors and host preferences influence \u003cem\u003eD. caninum\u003c/em\u003e infection (Liuet al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Consequently, even though males are more likely to develop an infection than females are, this difference is an insignificant (Fourie et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Similar to how infection varies with age, it actually increases slightly in younger animals (Trasvi\u0026ntilde;a-Mu\u0026ntilde;oz et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) due to older enhanced immune activity (East et al. 2103). In spite some studies, the prevalence is rising among the elderly (Cant\u0026oacute; et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe final host's lifestyle has an impact on the infection rate. Lack of care for stray and shelter animals causes infection rates to grow among them (Zottler et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) especially when intermediate hosts (Abdullah et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Because of cats' self-care behaviour, dogs may consume fewer fleas (Beugnet et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2014\u003c/span\u003e), but cats experience fewer flea infestations than dogs (Garc\u0026iacute;a-Agudo et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe hand lens or a microscope can be utilized to set eye on the worm, but a molecular diagnostic using specialised primers for specific genes yields more accurate and trustworthy results. While prior research were intended to identify the parasite in isolated flea samples (Low et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), Zhu et al. (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) confirmed the feasibility of recovering DNA from parasite eggs isolated from dog feces. Beugnet et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) revealed the presence of two unique sequences from 28S rDNA region when amplified a region DNA extracted from canine and feline fleas by PCR technique. He explained this by the existence of two distinct genotypes, each of which infects t flea species (\u003cem\u003eC. canis or C. felis\u003c/em\u003e) (Beugnet et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn line with this, Labuschagne et al. (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) created genotyping assays based on hydrolysis probes and restriction fragment length polymorphism (RFLP) analyses to identify and classify \u003cem\u003eD. caninum\u003c/em\u003e isolated from fleas. In order to find \u003cem\u003eD. caninum\u003c/em\u003e DNA in cat\u0026rsquo;s faeces, he used a mitochondrial genome. He did this to demonstrate that \u003cem\u003eD. caninum\u003c/em\u003e infections are highly particular to the host. He confirmed that the feline genotype was more common in cats than in dogs, with the canine genotype being more common in dogs (Labuschagne et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). In a similar manner, none of the gravid proglottids that have been isolated from feces samples exhibits any evidence of hybridization, despite the fact that various adult \u003cem\u003eD. caninum\u003c/em\u003e strains are capable of developing in the host intestine (Labuschagne et al. 2019).\u003c/p\u003e \u003cp\u003eIn Malaysia, it was established that the genotype of \u003cem\u003eD. caninum\u003c/em\u003e isolated from cats perfectly matches the DNA obtained from fleas on cats with tapeworm. They are two discrete clades that belong to two phenotypes that infect cats and dogs and are found in different parts of the world with differing morphological characteristics. They did not adhere to the regional distribution pattern or a particular geographic location (Low et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). A brand-new type of tapeworm known as \u003cem\u003eD. carracidoi\u003c/em\u003e was discovered in Spain. Its similarity to \u003cem\u003eD. caninum\u003c/em\u003e as determined by phenotypic research may indicate that it is a different genotype because of the latter species (Mill\u0026aacute;n and Casanova \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn conclusion, the gravid proglottids of worms isolated from definitive hosts can be used for molecular diagnosis and genotyping. \u003cem\u003eD. caninum\u003c/em\u003e exists in two different genotypes, one infecting cats and the other in dogs. The use of the RFLP approach and restriction enzymes in the three parasite positions (intestine, feces, and fleas) will probably give results that are more accurate.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eAll paper parts were done y Musafer AlArdi\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eAcknowledgement\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAl-Hamza Veterinary Hospital staff, especially Dr. Amin Abdulameer, I really appreciate your help.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe author published this article and has no direct interest in these studies other than publishing the scientific findings.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAbdullah S, Helps C, Tasker S, Newbury H, Wall R (2019) Pathogens in fleas collected from cats and dogs: Distribution and prevalence in the UK. 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Int J Chem Stud 7:550\u0026ndash;552. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.chemijournal.com/archives/2019/vol7issue1/PartJ/6-6-339-723.pdf\u003c/span\u003e\u003cspan address=\"https://www.chemijournal.com/archives/2019/vol7issue1/PartJ/6-6-339-723.pdf\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Dipylidium caninum, Genotyping, Felis catus, rRNA, tapeworm","lastPublishedDoi":"10.21203/rs.3.rs-4535872/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4535872/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eKnowing the parasites' genotypes and establishing their phylogenetic tree help to define their route of evolution and determine their origin. On the other hand, understanding the variation in genome sequences aids in the development of techniques for identifying diseases and selecting the best and most efficient treatments. This study used worms isolated from cats and dogs intestines to genotyping and draw phylogenetic tree of \u003cem\u003eDipylidium caninum\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eThis study approach involved isolating worms directly from the intestines of dogs and cats, and confirming infection through molecular diagnosis using primers designed by the author. Genotyping using primers for the large subunit rRNA gene revealed sequences of DNA products that were deposited in the GenBank. Finally, these sequences were aligned with previous sequences deposited in the GenBank, and gene sequences in cats were compared with their counterparts in dogs.\u003c/p\u003e \u003cp\u003eInfection significantly were higher among dogs than cats and among males than females. Three sequences (OQ987885 to OQ987887) isolated from dogs and another (OQ987888 to OQ987890) isolated from cats have been deposited in the Gen Bank. Phylogenetic tree comparing these sequences and showing the genetic distance between them. Sequences deposited from this study recorded high alignment with (MG587892.1) and (OK523385.1) sequences dating back to \u003cem\u003eD. caninum\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eThe host species influences the species and ability of the parasite that will infect it, and possibly the genotypes as well. Canine fleas can infect dogs and cats, while feline fleas can infect cat only. so the parasite must adapt to continue living, and one of the ways to adapt is heterogeneity of genotypes.\u003c/p\u003e","manuscriptTitle":"Phylogenetic study of Dipylidium caninum Isolated from cats (Felis domestica) and dogs (Canis familiaris) intestines.","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-25 18:19:19","doi":"10.21203/rs.3.rs-4535872/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"19972d89-ff41-4a0c-8145-bfb7f619b8e0","owner":[],"postedDate":"June 25th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-01-15T23:53:16+00:00","versionOfRecord":[],"versionCreatedAt":"2024-06-25 18:19:19","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4535872","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4535872","identity":"rs-4535872","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}