Molecular Detection of Novel Genetic Variants Associated to Anaplasma Ovis Among Dromedary Camels in Iran

Document Type: Original Articles

Author

Department of Veterinary Group of Animal Science Research, Isfahan Agricultural and Natural Resources Research and Education Center, Agricultural Research, Education, and Extension Organization, Isfahan, Iran

Abstract

To the best of our knowledge, little information is available regarding the presence of Anaplasma species in camels in Iran. This study sought to investigate the presence of Anaplasma species by microscopy and polymerase chain reaction (PCR) assays in 100 healthy dromedaries (Camelus dromedarius) arriving for slaughter. The microscopic examination of Giemsa-stained blood films revealed that Anaplasma like structures could be identified in the erythrocytes of two blood smears. To confirm the presence of and to identify the species of Anaplasma spp., a PCR technique was performed using primers amplifying a 750 bp fragment of the 16S rRNA gene of Anaplasma and the PCR products were analyzed by sequencing. The nucleotide sequence was compared to the sequences available in GenBank using the Basic Local Alignment Search Tool (BLAST). According to the results, the sequences of two 16S rRNA PCR products clearly fit within the Anaplasma genus in the family Anaplas mataceae. In this study, phylogenetic analysis using the 16S rRNA gene sequences revealed that two sequences obtained from monophyletic clusters included Anaplasma ovis (A. ovis). The obtained sequences had 99.6-100% similarity with previously published 16S rRNA gene sequences. This study aimed to evaluate the presence of novel genetic variants associated to A. ovis in dromedaries in the world. Further studies are recommended to establish the vector(s), as well as the veterinary and medical significance of these apparently novel variants in Iran.To the best of our knowledge, little information is available regarding the presence of Anaplasma species in camels in Iran. This study sought to investigate the presence of Anaplasma species by microscopy and polymerase chain reaction (PCR) assays in 100 healthy dromedaries (Camelus dromedarius) arriving for slaughter. The microscopic examination of Giemsa-stained blood films revealed that Anaplasma like structures could be identified in the erythrocytes of two blood smears. To confirm the presence of and to identify the species of Anaplasma spp., a PCR technique was performed using primers amplifying a 750 bp fragment of the 16S rRNA gene of Anaplasma and the PCR products were analyzed by sequencing. The nucleotide sequence was compared to the sequences available in GenBank using the Basic Local Alignment Search Tool (BLAST). According to the results, the sequences of two 16S rRNA PCR products clearly fit within the Anaplasma genus in the family Anaplas mataceae. In this study, phylogenetic analysis using the 16S rRNA gene sequences revealed that two sequences obtained from monophyletic clusters included Anaplasma ovis (A. ovis). The obtained sequences had 99.6-100% similarity with previously published 16S rRNA gene sequences. This study aimed to evaluate the presence of novel genetic variants associated to A. ovis in dromedaries in the world. Further studies are recommended to establish the vector(s), as well as the veterinary and medical significance of these apparently novel variants in Iran.

Keywords

Main Subjects


Article Title [French]

Détection moléculaire de nouvelles variantes génétiques associées à Anaplasma ovis chez les dromadaires iraniens (Camelus dromedarius)

Abstract [French]

Peu d'informations sont disponibles concernant la prévalence des infections anaplas miques chez les Camélidés iraniens. Une centaine de dromadaires (Camelus dromedarius) sains, arrivant à l'abattoir pour y être abattus, ont été examinés par microscopie et par des tests de réaction en chaîne de la polymérase (PCR) afin de détecter des contaminations anaplasmiques éventuelles. L'observation microscopique des frottis de sang périphérique colorés au Giemsa a révélé la présence de structures analogues à l’Anaplasma dans deux pourcent des échantillons sanguins. Afin de confirmer ces résultats et d’identifier les espèces d'Anaplasma impliquées, une technique de PCR a été mise au point en utilisant des amorces amplifiant un fragment de 750 pb du gène de l'ARNr 16S de l’Anaplasma. Les produits de cette PCR ont été ensuite analysés par séquençage. La séquence nucléotidique obtenue a été comparée aux séquences disponibles dans GenBank en utilisant le programme BLAST (Basic Local Alignment Search Tool). Ces analyses ont démontré que les séquences de deux produits de PCR ARNr 16S correspondent au genre Anaplasma appartenant à la famille des Anaplas mataceaes. L'analyse phylogénétique utilisant les séquences du gène de l'ARNr 16S classait les deux séquences obtenues dans cette étude dans des groupes monophylétiques comprenant A. ovis. Les résultats ont indiqué une similitude de 100 à 99,6% avec les séquences publiées du gène de l'ARNr 16S d'A. ovis. La présente étude rapporte la présence de nouvelles variantes génétiques associées à A. ovis chez les dromadaires dans le monde. Des recherches supplémentaires sont nécessaires afin d’identifier le ou les vecteur(s) de cette maladie, ainsi que l’impact de ces nouvelles variantes identifiées en Iran aux niveaux vétérinaire, médical et sanitaire.

Keywords [French]

  • Anaplasma ovis
  • dromadaire
  • identification moléculaire
  • gène de l'ARNr 16S
  • Iran

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