Muhammad Faiz RASOOL, Nisar AHMAD, Haroon AKBAR, Tahir YAQUB, Kamran ASHRAF, Aft ab Ahmad ANJUM, Mian Abdul HAFEEZ, Muhammad Muddassir ALI

Bm86 Genetic Diversity of Indigenous Tick Population from Punjab Province Pakistan

Ticks transmit a number of bacterial, protozoal and viral pathogens that cause many diseases like ehrlichiosis, hemorrhagic fever, theileriosis, babesiosis and anaplasmosis in livestock. This study was designed for molecular characterization of BM86 gene of Rhipicephalus (Boophilus) microplus tick. In this study, the BM86 gene was amplified, using primers fl anked by restriction enzyme sites. The molecular detection of R. microplus was studied in three districts belonging to diff erent ecological zones in the province of the Punjab, Pakistan. Tick samples were collected and initially screened through microscopy and further analyzed by PCR and sequencing. The phylogenetic tree was generated by using the MEGA 7 through Neighbor Joining method employing best model through the phylogenetic analysis of R. microplus. Pairwise comparisons of nucleotide sequences showed nucleotide diff erences ranging between 0.007 and 0.01%. Haplotype and nucleotide diversity in Bm86 gene was found among diff erent districts. Six single nucleotide polymorphisms were seen in sequences of BM 86 from indigenous tick populations collected from the Punjab province. More interestingly, out of these 6 polymorphisms, we got 2 from district Okara, 3 from district Sahiwal, and 1 in Mandi Bahauddin. Conserved regions were observed among the local strains for BM86 gene. A common convergence in similar clade was with the local Pakistan R. microplus. Local mean diversity was 0.005 and overall mean diversity was 0.038. Field strain has been isolated as candidate specie for local tick vaccine, which in turn will increase the efficacy of future tick vaccine including reduction of economic burden on the farmer.

Pakistan’ın Punjab Eyaletine Özgü Kene Popülasyonunun Bm86 Genetik Çeşitliliği

Keneler, çiftlik hayvanlarında erlişiyozis, hemorajik ateş, theileriozis, babeziozis ve anaplazmozis gibi birçok hastalığa neden olan bir dizi bakteriyel, protozoal ve viral patojeni bulaştırırlar. Bu çalışma, Rhipicephalus (Boophilus) microplus kenesinin BM86 geninin moleküler karakterizasyonu için tasarlanmıştır. Bu çalışmada, BM86 geni restriksiyon enzim bölgeleri ile çevrili primerler kullanılarak amplifiye edildi. R. microplus’un moleküler tespiti, Pakistan’ın Punjab eyaletindeki farklı ekolojik bölgelere ait üç yörede gerçekleştirildi. Kene örnekleri toplanıp ilk mikroskopik tarama yapıldıktan sonra, PCR ve sekans analizleri gerçekleştirildi. Filogenetik ağaç, R. microplus’un filogenetik analizi için en iyi model olan komşu birleştirme yöntemi kullanılarak MEGA 7 ile oluşturuldu. Nükleotid dizilerinin ikili karşılaştırmaları %0.007 ile %0.01 arasında değişen nükleotid farklılıkları olduğunu saptadı. Çalışılan farklı bölgelerde Bm86 geninde haplotip ve nükleotid çeşitliliği saptandı. Punjab eyaletinden toplanan lokal kene popülasyonunda BM86 geninde altı adet tek nükleotid polimorfizmi belirlendi. Daha da ilginci, bu 6 polimorfizmden 2’si Okara, 3’ü Sahiwal ve 1’i Mandi Bahauddin bölgesine aitti. Yerel suşlara ait BM86 geninde korunaklı bölgeler saptandı. Pakistan’ın lokal R. microplus suşlarında benzer sınıfta ortak bir kümelenme gözlendi. Lokal ortalama çeşitlilik 0.005 ve genel ortalama çeşitlilik 0.038 olarak saptandı. Lokal kene aşı aday türü olarak saha suşunun izolasyonu, çiftçiler üzerindeki ekonomik yükün azaltılması da dahil olmak üzere gelecekteki kene aşısının etkinliğini artıracaktır.

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