Abstract
The aim of this study was to estimate the molecular prevalence of Toxoplasma gondii, Neospora caninum and the co-infection by both Apicomplexan parasites in uterus tissues of cows. PCR was used to detect T. gondii and N. caninum DNA in uterus from 140 uteri of slaughtered cows in the regional slaughterhouse of Béja (Northwest Tunisia). Positive PCR products were sequenced and used for the phylogenetic analysis. The overall molecular prevalence of T. gondii and N. caninum in cows’ uterus was 5 and 15.57%, respectively. Co-infection prevalence by the two parasites was estimated to be 2.85%. Risk factors including the age categories significantly affected the molecular prevalence of T. gondii and N. caninum in cows’ uterus. The highest molecular prevalence of T. gondii (11.5 ± 3.1) and N. caninum (21.1 ± 11.1; p = 0.038) was observed in cows aged of more than 8 years. There were no differences depicted according to cow’s breeds and localities. Comparison of the partial sequences of the ITS1 gene revealed 100% similarity among our N. caninum sequence (MW136256) and those deposited in GenBank. The T. gondii sequence described in this study (MW260335) was 99.4–100% homologous to T. gondii sequences published in the GenBank.
To the best of our knowledge, this is the first molecular evidence of N. caninum and T. gondii co-infection in naturally infected cows in North Africa. This information is pertinent in designing control programmes that would reduce economic losses in the livestock industry.
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References
Almería, S., & López-gatius, F. (2013). Bovine neosporosis : clinical and practical aspects. Research in Veterinary Science, 95 (2), 303–309. 6. https://doi.org/10.1016/j.rvsc.2013.04.008
Amdouni, Y., Rjeibi, M. R., Awadi, S., Rekik, M., & Gharbi, M. (2018). First detection and molecular identification of Neospora caninum from naturally infected cattle and sheep in North Africa. Transboundary and Emerging Diseases 1–7. https://doi.org/10.1111/tbed.12828
Amdouni, Y., Rjeibi, M. R., Rouatbi, M., Amairia, S., Awadi, S., & Gharbi, M. (2017). Molecular detection of Toxoplasma gondii infection in slaughtered ruminants (sheep, goats and cattle) in Northwest Tunisia. Meat Science 133, 180–184. https://doi.org/10.1016/j.meatsci.2017.07.004
Amdouni, Y., Rouatbi, M., Lassoued, N., Rekik, M., & Gharbi, M. (2019). Neospora caninum natural infection in Tunisian rams: serological study and molecular identification of infection in semen. Acta Parasitologica 64, 821–828. https://doi.org/10.2478/s11686-019-00105-0
Ancelle, T. (2006). Statistique épidémiologie (2nd ed.). MALOINE. 300, 197-199.
Arbabi, M., Abdoli, A., Dalimi, A., & Pirestani, M. (2016). Identification of latent neosporosis in sheep in Tehran, Iran by polymerase chain reaction using primers specific for the Nc-5 gene. Onderstepoort Journal of Veterinary Research 83, 1–7. https://doi.org/10.4102/ojvr.v83i1.1058
Asgari, Q., & Sarnevesht, J. (2011). Molecular survey of Toxoplasma infection in sheep and goat from Fars province, Southern Iran. Tropical Animal Health and Production 43, 389–392. doi: https://doi.org/10.1007/s11250-010-9704-1
Berger-schoch, A.E., Herrmann, D.C., Schares, G., Müller, N., Bernet, D., Gottstein, B., & Frey, C.F. (2011). Prevalence and genotypes of Toxoplasma gondii in feline faeces (oocysts) and meat from sheep, cattle and pigs in Switzerland. Veterinary Parasitology 177, 290–297. https://doi.org/10.1016/j.vetpar.2010.11.046
Bishop, S., King, J., Windsor, P., Reichel, M.P., Ellis, J., & Jan, S. (2010). The first report of ovine cerebral neosporosis and evaluation of Neospora caninum prevalence in sheep in New South Wales. Veterinary Parasitology 170, 137–142. https://doi.org/10.1016/j.vetpar.2010.01.030
Boughattas, S., Ayari, K., Sa, T., Aoun, K., & Bouratbine, A. (2014). Survey of the parasite Toxoplasma gondii in Human consumed ovine meat in Tunis City. PLoS One 9, 1–5. https://doi.org/10.1371/journal.pone.0085044
Burrells, A., Bartley, P.M., Zimmer, I.A., Roy, S., Kitchener, A.C., Meredith, A., Wright, S.E., Innes, E.A., & Katzer, F. (2013). Evidence of the three main clonal Toxoplasma gondii lineages from wild mammalian carnivores in the UK. Parasitology 140, 1768–1776. https://doi.org/10.1017/S0031182013001169
Buxton, D., Maley, S.W., Wright, S., Thomson, K.M., Rae, A.G., & Innes, E.A. (1998). The pathogenesis of experimental neosporosis in pregnant sheep. Journal of Comparative Pathology 118, 267–279. https://doi.org/10.1016/S0021-9975(07)80003-X
Caetano-Da-Silva, A., Ferre, I., Collantes-Fernández, E., Navarro, V., Aduriz, G., Ugarte-Garagalza, C., and Ortega-Mora, L. M. (2004). Occasional detection of Neospora caninum DNA in frozen extended semen from naturally infected bulls. Theriogenology 62 (7), 1329–1336. https://doi.org/10.1016/j.theriogenology.2004.01.010
Cheadle, M.A., Spencer, J.A., & Blagburn, B.L., (1999). Seroprevalences of Neospora caninum and Toxoplasma gondii in nondomestic felids from southern Africa. Journal of Zoo and Wildlife Medicine 30, 248–251. https://www.jstor.org/stable/20095852
Cheah, T. S., Mattsson, J. G., Zaini, M., & Sani, R. A. (2004). Isolation of Neospora caninum from a calf in Malaysia. Veterinary Parasitology 126, 263–269. https://doi.org/10.1016/j.vetpar.2004.08.012
Cosendey-KezenLeite, R.I.J., de Oliveira, F.C.R., Frazão-Teixeira, E., Dubey, J.P., de Souza, G.N., Ferreira, A.M.R., & Lilenbaum, W. (2014). Occurrence and risk factors associated to Toxoplasma gondii infection in sheep from Rio de Janeiro, Brazil. Tropical Animal Health and Production 46, 1463–1466. https://doi.org/10.1007/s11250-014-0667-5
Du, L., Yang, D., Zhai, T., Gong, P., Zhang, X., & Li, J. (2015). Detection of Neospora caninum-DNA in brain tissues from pigeons in Changchun, Jilin (China). Veterinary Parasitology 214 (1–2), 171–173. https://doi.org/10.1016/j.vetpar.2015.09.005
Dubey, J.P., 1986. A review of toxoplasmosis in cattle. Veterinary Parasitology 22, 177–202. https://doi.org/10.1016/0304-4017(86)90106-8
Dubey, J. P., Hemphill, A., Calero-Bernal, R., & Schares, G. (2017). Neosporosis in animals. CRC Press, 530, 133-336
Dubey, J.P., Schares, G., & Ortega-Mora, L.M. (2007). Epidemiology and control of neosporosis and Neospora caninum. Clinical Microbiology Reviews 20, 323–367. https://doi.org/10.1128/CMR.00031-06
Ezatpour, B., Alirezaei, M., & Hassanvand, A. (2013). The first report of Neospora caninum prevalence in aborted and healthy sheep from west of Iran. Comparative Clinical Pathology 1–7https://doi.org/10.1007/s00580-013-1846-x
Ferre, I., Aduriz, G., Del-Pozo, I., Regidor-Cerrillo, J., Atxaerandio, R., Collantes- -Fernández, E., Ana, H., Carlos, U.G., & Ortega-Mora, L. M. (2005). Detection of Neospora caninum in the semen and blood of naturally infected bulls. Theriogenology 63 (5), 1504–1518. https://doi.org/10.1016/j.theriogenology.2004.07.002
Figliuolo, L P C, Kasai, N., Ragozo, A.M.A., Paula, V.S.O. De, Dias, R.A., Souza, S.L.P., & Gennari, S.M. (2004a). Prevalence of anti- Toxoplasma gondii and anti- Neospora caninum antibodies in ovine from São Paulo State, Brazil. Veterinary Parasitology 123, 161–166. https://doi.org/10.1016/j.vetpar.2004.06.006
Figliuolo, L. P C, Rodrigues, A.A.R., Viana, R.B., Aguiar, D.M., Kasai, N., & Gennari, S.M., (2004b). Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in goat from São Paulo State, Brazil. Small Ruminant Research 55, 29–32. https://doi.org/10.1016/j.smallrumres.2003.12.013
Franco-Hernandez, E.N., Acosta, A., Cortés-Vecino, J., & Gómez-Marín, J.E. (2016). Survey for Toxoplasma gondii by PCR detection in meat for human consumption in Colombia. Parasitology Research 115, 691–695. https://doi.org/10.1007/s00436-015-4790-7
Gharbi, M., Zribi, L., Jedidi, M., Chakkhari, H., Hamdi, S., R’hayem, S., Zribi, N., Souli, M., & Darghouth, M.A. (2013). Prévalence d’infection des ovins par Toxoplasma gondii en Tunisie. Bulletin de la Société de Pathologie Exotique 106, 184–187. https://doi.org/10.1007/s13149-013-0290-4
Ghoneim, N.H., Shalaby, S.I., Hassanain, N.A., Zeedan, G.S.G., Soliman, Y.A., & Abdalhamed, A.M. (2010). Comparative study between serological and molecular methods for diagnosis of toxoplasmosis in women and small ruminants in Egypt. Foodborne Pathogens and Disease 7, 17–22. https://doi.org/10.1089/fpd.2008.0223
Goodswen, S.J., Kennedy, P.J., & Ellis, J.T. (2013). A review of the infection, genetics, and evolution of Neospora caninum: from the past to the present. Infection, Genetics and Evolution 13, 133–150. https://doi.org/10.1016/j.meegid.2012.08.012
Gottstein, B., Hentrich, B., Wyss, R., Thür, B., Bruckner, L., Müller, N., Kaufmann, H., & Waldvogel, A. (1999). Molecular and immunodiagnostic studies of bovine neosporosis in Switzerland. International Journal of Parasitology 28 (4), 679-691. https://doi.org/10.1016/S0020-7519(98)00006-X
Hamilton, C.M., Gray, R., Wright, S.E., Gangadharan, B., Laurenson, K., & Innes, E.A. (2005). Prevalence of antibodies to Toxoplasma gondii and Neospora caninum in red foxes (Vulpes vulpes) from around the UK. Veterinary Parasitology 130, 169–173. https://doi.org/10.1016/j.vetpar.2005.03.020
Hässig, M., Sager, H., Reitt, K., Ziegler, D., Strabel, D., & Gottstein, B. (2003). Neospora caninum in sheep : a herd case report. Veterinary Parasitology 117, 213–220. https://doi.org/10.1016/j.vetpar.2003.07.029
Hilali, M., Romand, S., Thulliez, P., Kwok, O.C.H., & Dubey, J.P. (1998). Prevalence of Neospora caninum and Toxoplasma gondii antibodies in sera from camels from Egypt. Veterinary Parasitology 75, 269–271. https://doi.org/10.1016/S0304-4017(97)00181-7
Hughes, J.M., Thomasson, D., Craig, P.S., Georgin, S., Pickles, A., & Hide, G. (2008). Neospora caninum: Detection in wild rabbits and investigation of co-infection with Toxoplasma gondii by PCR analysis. Experimental Parasitology 120, 255–260. https://doi.org/10.1016/j.exppara.2008.07.011
Hughes, J.M., Williams, R.H., Morley, E.K., Cook, D. a N., Terry, R.S., Murphy, R.G., Smith, J.E., & Hide, G. (2006). The prevalence of Neospora caninum and co-infection with Toxoplasma gondii by PCR analysis in naturally occurring mammal populations. Parasitology 132, 29–36. https://doi.org/10.1017/S0031182005008784
Iovu, A., Györke, A., Mircean, V., Gavrea, R., & Cozma, V. (2012). Seroprevalence of Toxoplasma gondii and Neospora caninum in dairy goats from Romania. Veterinary Parasitology 186, 470–474. https://doi.org/10.1016/j.vetpar.2011.11.062
Leblebicier, A., & Yildiz, K. (2014). Seroprevalence of Toxoplasma gondii in sheep in Silopi district by using indirect fluorescent antibody test (IFAT). Türkiye Parazitoloji Dergisi 38, 1–4. https://doi.org/10.5152/tpd.2014.3329
Lindsay, D.S., Kelly, E.J., McKown, R.D., Stein, F.J., Plozer, J., Herman, J., Blagburn, B.L., & Dubey, J.P. (1996). Prevalence of Neospora caninum and Toxoplasma gondii antibodies in coyotes (Canis latrans) and experimental infections of coyotes with Neospora caninum. Journal of Parasitology 82, 657–659. https://doi.org/10.2307/3283798
Lopes, A.P., Vilares, A., Neto, F., Rodrigues, A., Martins, T., Ferreira, I., Gargaté, M.J., Rodrigues, M., Cardoso, L., & Article, O. (2015). Genoty** characterization of Toxoplasma gondii in cattle, sheep, goats and swine from the North of Portugal. Iranian Journal of Parasitology 10, 465–472. http://ijpa.tums.ac.ir
Opsteegh, M., Teunis, P., Mensink, M., Züchner, L., Titilincu, A., Langelaar, M., van der & Giessen, J. (2010). Evaluation of ELISA test characteristics and estimation of Toxoplasma gondii seroprevalence in Dutch sheep using mixture models. Preventive Veterinary Medicine 96, 232–240. https://doi.org/10.1016/j.prevetmed.2010.06.009
Ortega-Mora, L. M., Ferre, I., Del-Pozo, I., Caetano-Da-Silva, A., Collantes- Fernández, E., Regidor-Cerrillo, J., Ugarte-Garagalza, C., & Aduriz, G. (2003). Detection of Neospora caninum in semen of bulls. Veterinary Parasitology 117(4), 301–308. https://doi.org/10.1016/j.vetpar.2003.09.015
Pan, Y., Jansen, G.B., Duffield, T.F., Hietala, S., Kelton, D., Lin, C.Y., & Peregrine, A.S. (2004). Genetic susceptibility to Neospora caninum infection in Holstein cattle in Ontario. Journal of Dairy science 87, 3967–3975. https://doi.org/10.3168/jds.S0022-0302(04)73537-7
Perry, B.D., Grieve, A.S., Mogollon, J.D., & de Galvis, A.L. (1978). Serological study of ovine toxoplasmosis in Colombia: prevalence of haemagglutinating antibodies to toxoplasma in sheep. Veterinary Record 103, 584–585. https://doi.org/10.1136/vr.103.26-27.584
Reichel, M.P., Alejandra Ayanegui-Alcérreca, M., Gondim, L.F.P., & Ellis, J.T. (2013). What is the global economic impact of Neospora caninum in cattle — the billion dollar question. International Journal of Parasitology 43, 133–142. https://doi.org/10.1016/j.ijpara.2012.10.022
Reid, A.J., Vermont, S.J., Cotton, J.A., Harris, D., Hill-Cawthorne, G.A., Könen-Waisman, S., Latham, S.M., Mourier, T., Norton, R., Quail, M.A., Sanders, M., Shanmugam, D., Sohal, A., Wasmuth, J.D., Brunk, B., Grigg, M.E., Howard, J.C., Parkinson, J., Roos, D.S., Trees, A.J., Berriman, M., Pain, A., & Wastling, J.M. (2012). Comparative genomics of the apicomplexan parasites Toxoplasma gondii and Neospora caninum: Coccidia differing in host range and transmission strategy. PLoS Pathogens 8. https://doi.org/10.1371/journal.ppat.1002567
Santos, S.L., De Souza Costa, K., Gondim, L.Q., Da Silva, M.S.A., Uzêda, R.S., Abe-Sandes, K., & Gondim, L.F.P. (2010). Investigation of Neospora caninum, Hammondia sp., and Toxoplasma gondii in tissues from slaughtered beef cattle in Bahia, Brazil. Parasitology Research 106, 457–461. https://doi.org/10.1007/s00436-009-1686-4
Samra N.A., McCrindle C.M.E., Penzhorn B.L.& Cenci-Goga B. (2007). Seroprevalence of toxoplasmosis in sheep in South Africa. Journal of the South African Veterinary Association, 78, 116–120. https://doi.org/10.4102/jsava.v78i3.301
Sasani, F., Javanbakht, J., Seifori, P., Fathi, S., Hassan, M.A. (2013). Neospora caninum as causative agent of ovine encephalitis in Iran. Pathology Discovery 3–7https://doi.org/10.7243/2052-7896-1-5
Schwartz D. 1993: Méthodes statistiques à l’usage des médecins et des biologistes, 3ème éd, Flammarion: Paris, France. 83: 170-181
Sundermann, C.A., & Estridge, B.H. (1999). Growth of and competition between Neospora caninum and Toxoplasma gondii in vitro. International Journal of Parasitology 29, 1725–1732. https://doi.org/10.1016/S0020-7519(99)00114-9
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., & Kumar, S. (2011). MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution 28, 2731–2739. https://doi.org/10.1093/molbev/msr121
Thrusfield, M. V. (2005). Veterinary epidemiology (2nd ed.). Blackwell Science.
Wang, Y., Tian, R.M., Gao, Z.M., Bougouffa, S., & Qian, P.Y. (2014). Optimal eukaryotic 18S and universal 16S/18S ribosomal RNA primers and their application in a study of symbiosis. PLoS One 9. https://doi.org/10.1371/journal.pone.0090053
Yanina P. H., Moore, D. P., Manazza, J. A., Unzaga, J. M., Späth, E. J. A., Pardini, L. L., Venturini, M. C., Roberi, J. L., & Campero, C. M. (2013). First report of seroprevalence of Toxoplasma gondii and Neospora caninum in dairy sheep from Humid Pampa, Argentina. Tropical Animal Health and Production 45, 1645–1647. https://doi.org/10.1007/s11250-013-0396-1
Zhang, W., Deng, C., Liu, Q., Liu, J., Wang, M., Tian, K. G., X.L. Yu., & Hu, D. M. (2007). First identification of Neospora caninum infection in aborted bovine foetuses in China. Veterinary Parasitology 149, 72–76. https://doi.org/10.1016/j.vetpar.2007.07.013
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The authors thank Mr. Limam Sassi and Mr. Taoufik Lahmar for their technical support.
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The work was funded by the laboratory of “Laboratoire d’épidémiologie des infections enzootiques des herbivores en Tunisie: application à la lutte’’ (LR16AGR01) (Ministère de l’enseignement supérieure et de la recherche scientifique, Tunisie).
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AM wrote the manuscript and conducted experiments, MRJ did the phylogenetic analysis, AS collected the samples, GM revised the manuscript. All authors read and approved the final manuscript.
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Amdouni, Y., Rjeibi, M.R., Awadi, S. et al. Molecular identification of Neospora caninum and co-infection with Toxoplasma gondii in genital apparatus of naturally infected cows in North Tunisia. Trop Anim Health Prod 53, 528 (2021). https://doi.org/10.1007/s11250-021-02975-x
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DOI: https://doi.org/10.1007/s11250-021-02975-x