Acta Parasitologica

, Volume 64, Issue 4, pp 821–828 | Cite as

Neospora caninum Natural Infection in Tunisian Rams: Serological Study and Molecular Identification of Infection in Semen

  • Yosra AmdouniEmail author
  • Mariem Rouatbi
  • Narjess Lassoued
  • Mourad Rekik
  • Mohamed Gharbi
Original Paper



The aim of this study was to estimate the seroprevalence and molecular prevalence and perform a molecular identification of Neospora caninum in semen of Tunisian rams.


A total of 92 blood samples were collected from four farms located in four Tunisian governorates (Jendouba, Kairouan, Zaghouan and Ben Arous) and samples were screened with a commercial ELISA kit for N. caninum antibodies. For the same rams, semen samples were collected and tested for the presence of N. caninum ITS1 gene using PCR. Five amplicons were randomly selected for sequencing. A phylogenetic tree was constructed to compare the partial sequences of the ITS1 gene with sequences deposited in GenBank.


The seroprevalence of N. caninum infection was 25% (23/92) and PCR revealed that the molecular infection prevalence in semen was 11.95% (11/92). Kappa test showed an average agreement between seroprevalence and parasite prevalence in semen (κ = 0.44). The highest molecular prevalence was for rams that accomplished more than two mating seasons (21.0 ± 12.1%) compared to those performed less than two mating seasons and yearling individuals (4.0 ± 5.5%) (P = 0.01). There were no differences in N. caninum molecular prevalence according to either breed or locality. Comparison of the partial sequences of the ITS1 gene revealed 99–100% similarity with those deposited in GenBank.


To the best of our knowledge, this is the first detection and molecular identification of N. caninum in semen from rams in North Africa. Our findings indicate that N. caninum infection rate was high in rams.


Neospora caninum infection Ram Semen PCR ELISA Tunisia 



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) and was partly supported by the CGIAR Research Program on Livestock. The authors thank Mr. Bechir Guesmi, Mr. Limam Sassi and Mr. Taoufik Lahmar for their technical support and the farm owners. We also acknowledge Mustapha Lahmer from the Department of Animal Breeding, Office de l’Elevage et des Pâturages, Sidi Thabet, Tunisia.

Compliance with Ethical Standards

Conflict of Interest

The authors declare no conflicts of interest in relation of this work.


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Copyright information

© Witold Stefański Institute of Parasitology, Polish Academy of Sciences 2019

Authors and Affiliations

  1. 1.Laboratoire de ParasitologieUniv. Manouba, Institution de la Recherche et de l’Enseignement Supérieur Agricoles, Ecole Nationale de Médecine Vétérinaire de Sidi ThabetSidi ThabetTunisia
  2. 2.Laboratory of Animal Production and ForagesNational Institute of Agronomic Research of Tunisia (INRAT), Carthage UniversityTunisTunisia
  3. 3.International Center for Agricultural Research in the Dry Areas (ICARDA)AmmanJordan

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