Molecular detection of Dirofilaria immitis and its Wolbachia endosymbionts in dogs from Myanmar

Saw Bawm , Yadanar Khaing , Hla Myet Chel , Myint Myint Hmoon , Shwe Yee Win , Min Bo , Tint Naing , Lat Lat Htun
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Abstract

Heartworm disease in dogs and cats caused by Dirofilaria immitis continues to be a major clinical issue globally. This study focused on dogs suspicious of having tick-borne diseases (TBD) brought to a clinic and a veterinary teaching hospital in Myanmar. Blood samples were collected and initially screened using SNAP® 4Dx® Plus test kit. All dog blood samples were subjected to conventional PCR to detect both Dirofilaria spp. (cox1 gene) and Wolbachia spp. (16S rDNA) infections. Infection with D. immitis was detected in 14 (28.0%) of 50 examined samples, while the detection rate of TBD causative agents, including Anaplasma phagocytophilum and Ehrlichia canis, was 26.0% (13/50) and 26.0% (13/50), respectively, as determined by ELISA rapid test. In this study, D. immitis infection was moderately but significantly correlated with TBD infections (Pearson’s r = 0.397, P = 0.008). Comparative sequence and phylogenetic analyses provided molecular identification of D. immitis in Myanmar and confirmed the identity of its Wolbachia endosymbiont with Wolbachia endosymbionts isolated from D. immitis, Rhipicephalus sanguineus and Aedes aegypti. The present study contributes to our understanding of the coexistence of D. immitis and Wolbachia endosymbiosis in dogs, and the findings may benefit the future prevention and control of dirofilariasis in dogs.

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缅甸犬免疫dirofilia imimtis及其Wolbachia内共生菌的分子检测
狗和猫心丝虫引起的心丝虫病仍然是全球的一个主要临床问题。这项研究的重点是被带到缅甸一家诊所和兽医教学医院怀疑患有蜱传疾病的狗。采集血液样本,并使用SNAP®4Dx®Plus检测试剂盒进行初步筛选。采用常规PCR检测犬血中dirofilia spp (cox1基因)和Wolbachia spp (16S rDNA)感染。50份检测样本中有14份(28.0%)检出免疫弓形虫感染,ELISA快速检测TBD病原体嗜吞噬细胞无原体和犬埃利希体检出率分别为26.0%(13/50)和26.0%(13/50)。本研究中,D. immitis感染与TBD感染呈中度但显著相关(Pearson’s r = 0.397, P = 0.008)。序列比较和系统发育分析证实了缅甸金米特布氏菌的分子鉴定,并证实了其与金米特布氏菌、血根头虫和埃及伊蚊分离的沃尔巴克氏菌内生共生体同源性。本研究有助于我们对犬犬免疫弓形虫病和沃尔巴克氏体内共生共存的认识,并为今后犬双丝虫病的预防和控制提供参考。
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