{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T05:35:54Z","timestamp":1789191354912,"version":"build-2803163510"},"reference-count":54,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2020,10,28]],"date-time":"2020-10-28T00:00:00Z","timestamp":1603843200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>The zoonotic Onchocerca lupi and tick-transmitted filarioids of the genus Cercopithifilaria remain less well known due to the difficulties in accessing to skin samples as target tissues. Here, we proposed a molecular approach reliying on multiplex qPCR assays that allow the rapid identification of filarioids from canine blood, skin, and tick samples. This includes two newly developed duplex qPCR tests, the first one targeting filarial and C. grassii DNA (CanFil-C. grassii). and the second qPCR assay designed for the detection of Cercopithifilaria bainae and Cercopithifilaria sp. II DNAs (C. bainae-C.spII). The third one is a triplex TaqMan cox 1 assay targeting DNA of blood microfilariae (e.g., Dirofilaria immitis, Dirofilaria repens and Acanthocheilonema reconditum). The novel duplex qPCRs developed were validated in silico and by screening of known DNA collection. The qPCR assays were also used for screening the blood and tick samples of 72 dogs from Algeria. This allowed the identification of canine filariasis infection with 100% of specificity and 89.47% and 100% of sensitivity from naturally infected blood and tick samples, respectively. The prevalences of 26.39% for D. immitis and 5.56% for both D. repens and A. reconditum were reported in blood and tick samples. Cercopithifilaria DNAs were detected only in tick samples, with a prevalence of 4.17% and 5.56% for C. bainae and Cercopithifilaria sp. II, respectively. Co-infections were diagnosed in 6.94% and 13.89% of blood and tick samples, respectively. Whereas all samples were negative for C. grassii DNA. The use of engorged ticks instead of blood and skin samples could be an easier option for the surveillance of all canine filarioids herein investigated. The multiplex qPCR assays herein validated were shown to be useful in the detection of filarial co-infections by overcoming sequencing of positive samples.<\/jats:p>","DOI":"10.3390\/microorganisms8111671","type":"journal-article","created":{"date-parts":[[2020,10,28]],"date-time":"2020-10-28T11:43:06Z","timestamp":1603885386000},"page":"1671","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Molecular Approach for the Diagnosis of Blood and Skin Canine Filarioids"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8819-3019","authenticated-orcid":false,"given":"Younes","family":"Laidoudi","sequence":"first","affiliation":[{"name":"IRD, AP-HM, Aix Marseille Univ, 19-21 Boulevard Jean Moulin, 13005 Marseille, France"},{"name":"Microbes, Evolution, Phylogeny and Infection (MEPHI), IHU M\u00e9diterran\u00e9e Infection, 19-21 Boulevard Jean Moulin, 13005 Marseille, France"},{"name":"PADESCA Laboratory, Veterinary Science Institute, University Constantine 1, El Khroub 25100, Algeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samia","family":"Bedjaoui","sequence":"additional","affiliation":[{"name":"Laboratory of Food Hygiene and Quality Insurance System (HASAQ), Higher National Veterinary School, Issad Abbes, Oued Smar, Algiers 16000, Algeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5208-2576","authenticated-orcid":false,"given":"Hac\u00e8ne","family":"Medkour","sequence":"additional","affiliation":[{"name":"IRD, AP-HM, Aix Marseille Univ, 19-21 Boulevard Jean Moulin, 13005 Marseille, France"},{"name":"Microbes, Evolution, Phylogeny and Infection (MEPHI), IHU M\u00e9diterran\u00e9e Infection, 19-21 Boulevard Jean Moulin, 13005 Marseille, France"},{"name":"PADESCA Laboratory, Veterinary Science Institute, University Constantine 1, El Khroub 25100, Algeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9061-437X","authenticated-orcid":false,"given":"Maria Stefania","family":"Latrofa","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Bari, 70010 Valenzano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abdeslam","family":"Mekroud","sequence":"additional","affiliation":[{"name":"PADESCA Laboratory, Veterinary Science Institute, University Constantine 1, El Khroub 25100, Algeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Idir","family":"Bitam","sequence":"additional","affiliation":[{"name":"IRD, AP-HM, SSA, VITROME, Aix-Marseille Univ, 13385 Marseille, France"},{"name":"Superior School of Food Sciences and Food Industries, Algiers 16004, Algeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7836-6809","authenticated-orcid":false,"given":"Bernard","family":"Davoust","sequence":"additional","affiliation":[{"name":"IRD, AP-HM, Aix Marseille Univ, 19-21 Boulevard Jean Moulin, 13005 Marseille, France"},{"name":"Microbes, Evolution, Phylogeny and Infection (MEPHI), IHU M\u00e9diterran\u00e9e Infection, 19-21 Boulevard Jean Moulin, 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7518-476X","authenticated-orcid":false,"given":"Domenico","family":"Otranto","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine, University of Bari, 70010 Valenzano, Italy"},{"name":"Department of Pathobiology, Faculty of Veterinary Science, Bu-Ali Sina University, Felestin Sq, 6517658978 Hamedan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6039-2008","authenticated-orcid":false,"given":"Oleg","family":"Mediannikov","sequence":"additional","affiliation":[{"name":"IRD, AP-HM, Aix Marseille Univ, 19-21 Boulevard Jean Moulin, 13005 Marseille, France"},{"name":"Microbes, Evolution, Phylogeny and Infection (MEPHI), IHU M\u00e9diterran\u00e9e Infection, 19-21 Boulevard Jean Moulin, 13005 Marseille, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,10,28]]},"reference":[{"key":"ref_1","unstructured":"Anderson, R.C. (1992). Nematode Parasites of Vertebrates: Their Development and Transmission, C.A.B. International."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1155\/2014\/630160","article-title":"Canine filarial infections in a human Brugia malayi endemic area of India","volume":"2014","author":"Ravindran","year":"2014","journal-title":"BioMed Res. Int."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1371\/journal.pntd.0006402","article-title":"A real-time PCR tool for the surveillance of zoonotic Onchocerca lupi in dogs, cats and potential vectors","volume":"12","author":"Latrofa","year":"2018","journal-title":"PLoS Negl. Trop. Dis."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Otranto, D., Brianti, E., Abramo, F., Gaglio, G., Napoli, E., Latrofa, M.S., Ramos, R.A., Dantas-Torres, F., and Bain, O. (2012). Cutaneous distribution and localization of Cercopithifilaria sp. microfilariae in dogs. Vet. Parasitol.","DOI":"10.1016\/j.vetpar.2012.05.016"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13071-020-04185-0","article-title":"Development of a multiplex qPCR-based approach for the diagnosis of Dirofilaria immitis, D. repens and Acanthocheilonema reconditum","volume":"13","author":"Laidoudi","year":"2020","journal-title":"Parasites Vectors"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.vetpar.2007.09.008","article-title":"Onchocercosis: A newly recognized disease in dogs","volume":"151","year":"2008","journal-title":"Vet. Parasitol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1017\/S0031182012001357","article-title":"Species diversity of dermal microfilariae of the genus Cercopithifilaria infesting dogs in the Mediterranean region","volume":"140","author":"Otranto","year":"2013","journal-title":"Parasitology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1756-3305-7-261","article-title":"Diversity of Cercopithifilaria species in dogs from Portugal","volume":"7","author":"Cortes","year":"2014","journal-title":"Parasites Vectors"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"778","DOI":"10.1093\/cid\/civ983","article-title":"The emergence of zoonotic Onchocerca lupi infection in the United States\u2014A case-series","volume":"62","author":"Cantey","year":"2016","journal-title":"Clin. Infect. Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1756-3305-5-1","article-title":"On a Cercopithifilaria sp. transmitted by Rhipicephalus sanguineus: A neglected, but widespread filarioid of dogs","volume":"5","author":"Otranto","year":"2012","journal-title":"Parasites Vectors"},{"key":"ref_11","first-page":"715","article-title":"A new species of Nematode, Onchocerca lupi n. sp., from Canis lupus cubanensis","volume":"45","author":"Rodonaja","year":"1967","journal-title":"Soobscenija Akademii Nauk Gruzinskoj SSR"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13071-015-0699-3","article-title":"Clinical case presentation and a review of the literature of canine onchocercosis by Onchocerca lupi in the United States","volume":"8","author":"Otranto","year":"2015","journal-title":"Parasites Vectors"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1136\/vr.156.14.450","article-title":"First autochthonous case of canine ocular onchocercosis in Germany","volume":"156","author":"Hermosilla","year":"2005","journal-title":"Vet. Rec."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1016\/S0304-4017(01)00397-1","article-title":"Canine ocular onchocercosis in Hungary","volume":"97","author":"Albert","year":"2001","journal-title":"Vet. Parasitol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Labelle, A.L., Daniels, J.B., Dix, M., and Labelle, P. (2011). Onchocerca lupi causing ocular disease in two cats. Vet. Ophthalmol., 14.","DOI":"10.1111\/j.1463-5224.2011.00911.x"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Labelle, A.L., Maddox, C.W., Daniels, J.B., Lanka, S., Eggett, T.E., Dubielzig, R.R., and Labelle, P. (2013). Canine ocular onchocercosis in the United States is associated with Onchocerca lupi. Vet. Parasitol., 193.","DOI":"10.1016\/j.vetpar.2012.12.002"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2252","DOI":"10.3201\/eid2112.150061","article-title":"Onchocerca lupi nematode in cat, Portugal","volume":"21","author":"Maia","year":"2015","journal-title":"Emerg. Infect. Dis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2000","DOI":"10.3201\/eid1912.130264","article-title":"Zoonotic Onchocerca lupi infection in dogs, Greece and Portugal, 2011\u20132012","volume":"19","author":"Otranto","year":"2013","journal-title":"Emerg. Infect. Dis."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Mir\u00f3, G., Montoya, A., Checa, R., G\u00e1lvez, R., M\u00ednguez, J.J., Marino, V., and Otranto, D. (2016). First detection of Onchocerca lupi infection in dogs in southern Spain. Parasites Vectors, 9.","DOI":"10.1186\/s13071-016-1587-1"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Otranto, D., Dantas-Torres, F., Giannelli, A., Abramo, F., Ignjatovi\u0107 \u0106upina, A., Petri\u0107, D., Cardoso, L., Mutafchiev, Y., and Cortes, H. (2013). Cutaneous Distribution and Circadian Rhythm of Onchocerca lupi Microfilariae in Dogs. PLoS Negl. Trop. Dis.","DOI":"10.1371\/journal.pntd.0002585"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Bain, O., Aeschlimann, A., and Chatelanat, P. (1982). Pr\u00e9sence, chez des tiques de la r\u00e9gion de Gen\u00e8ve, de larves infestantes qui pourraient se rapporter \u00e0 la filaire de chien Dipetalonema grassii. Ann. Parasitol. Hum. Compar\u00e9e.","DOI":"10.1051\/parasite\/1982576643"},{"key":"ref_22","first-page":"316","article-title":"Ritrovamento di Diptalonema grassii (No\u00e8, 1907) in Rhipicephalus sanguineus su cane in Italia e descrizione di alcuni suoi stadi larvali","volume":"25","author":"Pampiglione","year":"1983","journal-title":"Parassitologia"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Otranto, D., Varcasia, A., Solinas, C., Scala, A., Brianti, E., Dantas-Torres, F., Annoscia, G., Martin, C., Mutafchiev, Y., and Bain, O. (2013). Redescription of Cercopithifilaria bainae Almeida & Vicente, 1984 (Spirurida, Onchocercidae) from a dog in Sardinia, Italy. Parasites Vectors.","DOI":"10.1186\/1756-3305-6-132"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Otranto, D., Latrofa, M.S., Brianti, E., Annoscia, G., Parisi, A., Dantas-Torres, F., Bain, O., and Gasser, R.B. (2012). An assessment of genetic variability in the mitochondrial cytochrome c oxidase subunit 1 gene of Cercopithifilaria sp. (Spirurida, Onchocercidae) from dog and Rhipicephalus sanguineus populations. Mol. Cell. Probes.","DOI":"10.1016\/j.mcp.2011.12.002"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Solinas, C., Varcasia, A., Brianti, E., Giannetto, S., Pipia, A.P., Columbano, N., Tosciri, G., Dantas-Torres, F., Garippa, G., and Otranto, D. (2014). Cercopithifilaria spp. in dogs in Sardinia Island (Italy). Parasitol. Res.","DOI":"10.1007\/s00436-013-3695-6"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1742-9994-6-1","article-title":"Integrated taxonomy: Traditional approach and DNA barcoding for the identification of filarioid worms and related parasites (Nematoda)","volume":"6","author":"Ferri","year":"2009","journal-title":"Front. Zool."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"SantaLucia, J. (1998). A unified view of polymer, dumbbell, and oligonucleotide DNA nearest-neighbor thermodynamics. Proc. Natl. Acad. Sci. USA.","DOI":"10.1073\/pnas.95.4.1460"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Ye, J., Coulouris, G., Zaretskaya, I., Cutcutache, I., Rozen, S., and Madden, T.L. (2012). Primer-BLAST: A tool to design target-specific primers for polymerase chain reaction. BMC Bioinform., 13.","DOI":"10.1186\/1471-2105-13-134"},{"key":"ref_29","unstructured":"Bio-Rad Lab (2006). Real-Time PCR Applications Guide, Bio-Rad Lab. Inc."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Medkour, H., Laidoudi, Y., Lafri, I., Davoust, B., Mekroud, A., Bitam, I., and Mediannikov, O. (2020). Canine vector-borne protozoa: Molecular and serological investigation for Leishmania spp., Trypanosoma spp., Babesia spp., and Hepatozoon spp. in dogs from Northern Algeria. Vet. Parasitol. Reg. Stud. Rep.","DOI":"10.1016\/j.vprsr.2019.100353"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.cimid.2016.01.002","article-title":"Molecular evidence of vector-borne pathogens in dogs and cats and their ectoparasites in Algiers, Algeria","volume":"45","author":"Bessas","year":"2016","journal-title":"Comp. Immunol. Microbiol. Infect. Dis."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Halos, L., Jamal, T., Vial, L., Maillard, R., Suau, A., Le Menach, A., Boulouis, H.J., and Vayssier-Taussat, M. (2004). Determination of an efficient and reliable method for DNA extraction from ticks. Vet. Res., 35.","DOI":"10.1051\/vetres:2004038"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Katoh, K., Misawa, K., Kuma, K.I., and Miyata, T. (2002). MAFFT: A novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res., 30.","DOI":"10.1093\/nar\/gkf436"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Tamura, K., Stecher, G., Peterson, D., Filipski, A., and Kumar, S. (2013). MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol.","DOI":"10.1093\/molbev\/mst197"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Letunic, I., and Bork, P. (2019). Interactive Tree of Life (iTOL) v4: Recent updates and new developments. Nucleic Acids Res., 47.","DOI":"10.1093\/nar\/gkz239"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"579","DOI":"10.2307\/2340126","article-title":"On the methods of measuring association between two attributes","volume":"75","author":"Yule","year":"1912","journal-title":"J. R. Stat. Soc."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1177\/0049124106292358","article-title":"Statistical inference for generalized Yule coefficients in 2 \u00d7 2 contingency tables","volume":"35","author":"Bonett","year":"2007","journal-title":"Sociol. Methods Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"777","DOI":"10.1007\/s11336-008-9070-3","article-title":"On association coefficients for 2 \u00d7 2 tables and properties that do not depend on the marginal distributions","volume":"73","author":"Warrens","year":"2008","journal-title":"Psychometrika"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1756-3305-6-16","article-title":"Vector-borne helminths of dogs and humans in Europe","volume":"6","author":"Otranto","year":"2013","journal-title":"Parasites Vectors"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Dantas-Torres, F., and Otranto, D. (2020). Overview on Dirofilaria immitis in the Americas, with notes on other filarial worms infecting dogs. Vet. Parasitol., 282.","DOI":"10.1016\/j.vetpar.2020.109113"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1016\/j.pt.2009.01.003","article-title":"Managing canine vector-borne diseases of zoonotic concern: Part one","volume":"25","author":"Otranto","year":"2009","journal-title":"Trends Parasitol."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Lineberry, M.W., Sundstrom, K.D., Little, S.E., Stayton, E.M., and Allen, K.E. (2020). Detection of Cercopithifilaria bainae infection in shelter dogs and ticks in Oklahoma, USA. Parasites Vectors, 13.","DOI":"10.1186\/s13071-020-04089-z"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Laidoudi, Y., Mari, J., Tahir, D., and Watier-grillot, S. (2020). Detection of Canine Vector-Borne Filariasis and Their Wolbachia Endosymbionts in French Guiana. Microorganisms, 8.","DOI":"10.3390\/microorganisms8050770"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"De Almeida, M., Nascimento, F.S., Mathison, B.A., Bishop, H., Bradbury, R.S., Cama, V.A., and da Silva, A.J. (2020). Duplex Real-Time PCR Assay for Clinical Differentiation of Onchocerca lupi and Onchocerca volvulus. Am. J. Trop. Med. Hyg.","DOI":"10.4269\/ajtmh.20-0113"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s13071-015-0783-8","article-title":"Detection of Dirofilaria immitis and other arthropod-borne filarioids by an HRM real-time qPCR, blood-concentrating techniques and a serological assay in dogs from Costa Rica","volume":"8","author":"Rojas","year":"2015","journal-title":"Parasites Vectors"},{"key":"ref_46","first-page":"5","article-title":"Filarial nematode infection in Ixodes scapularis ticks collected from Southern Connecticut","volume":"1","author":"Namrata","year":"2014","journal-title":"Vet. Sci."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Beaver, P.C., and Burgdorfer, W. (1984). A microfilaria of exceptional size from the ixodid tick, Ixodes dammini, from Shelter Island, New York. J. Parasitol., 70.","DOI":"10.2307\/3281647"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Voigt, C.C., Peschel, U., Wibbelt, G., and Fr\u00f6lich, K. (2006). An alternative, less invasive blood sample collection technique for serologic studies utilizing Triatomine bugs (Heteroptera; Insecta). J. Wildl. Dis., 42.","DOI":"10.7589\/0090-3558-42.2.466"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1186\/1756-3305-3-30","article-title":"A survey of canine filarial diseases of veterinary and public health significance in India","volume":"3","author":"Irwin","year":"2010","journal-title":"Parasites Vectors"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1017\/S0022149X11000113","article-title":"Canine filarial infections in Tuscany, central Italy","volume":"86","author":"Magi","year":"2012","journal-title":"J. Helminthol."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Latrofa, M.S., Dantas-Torres, F., Giannelli, A., and Otranto, D. (2014). Molecular detection of tick-borne pathogens in Rhipicephalus sanguineus group ticks. Ticks Tick Borne Dis., 5.","DOI":"10.1016\/j.ttbdis.2014.07.014"},{"key":"ref_52","first-page":"1097","article-title":"Prevalence of canine Dirofilaria immitis infection in the city of Algiers, Algeria","volume":"4","author":"Mohamed","year":"2009","journal-title":"Afr. J. Agric. Res."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Tahir, D., Damene, H., Davoust, B., and Parola, P. (2017). First molecular detection of Dirofilaria immitis (Spirurida: Onchocercidae) infection in dogs from Northern Algeria. Comp. Immunol. Microbiol. Infect. Dis., 51.","DOI":"10.1016\/j.cimid.2017.04.001"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Henning, T.C., Orr, J.M., Smith, J.D., Arias, J.R., Rasgon, J.L., and Norris, D.E. (2016). Discovery of filarial nematode DNA in Amblyomma americanum in Northern Virginia. Ticks Tick Borne Dis., 7.","DOI":"10.1016\/j.ttbdis.2015.11.007"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/8\/11\/1671\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:29:53Z","timestamp":1760178593000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/8\/11\/1671"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,10,28]]},"references-count":54,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2020,11]]}},"alternative-id":["microorganisms8111671"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms8111671","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,10,28]]}}}