Molecular identification of trypanosoma species infecting dogs in nigeria by analysis of partial internal transcribed spacer-1 of RRNA gene

P. U. Umeakuana, O. A. Adepoju, M. Takeet, G. Chechet, Lamin B. S. Dibba, R. Ezeokonkwo, B.M. Anene, E. Balogun
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Abstract

Trypanosomosis is becoming a major health challenge to dogs in the sub-Sahara Africa including Nigeria, but minimal reports are available on the  molecular characteristic of trypanosomes infecting dogs in Nigeria. We characterized trypanosomes detected in naturally infected dogs by PCR and  sequences analysis of the partial region of the trypanosomes internal transcribed spacer-1of ribosomal RNA (ITS1 rRNA). Animals presented to the  University of Nigeria Veterinary Teaching Hospital (UNVTH) in Nsukka, Nigeria for examination and treatment were sampled for laboratory tests. DNA was  extracted from 21 blood samples obtained from dogs that were confirmed positive for trypanosome infection by microscopy. ITS-1 was PCR  amplified and sequenced bi-directionally. Sixteen samples have good bands, though one of them had unreadable sequence. Analyses of the sequence  data by BLAST search on NCBI identified T. congolense, T. brucei gambiense, and T. evansi in 4.8, 4.8, and 91.4%, respectively, from the analysed samples  from the infected dogs. Although the top BLAST hits for T. brucei group were due to T. evansi and T. b. gambiense, there is not enough discriminatory  power in ITS-1 to distinguish subspecies. The aligned sequences of the trypanozoon were less polymorphic. Phylogenetic trees inferred by unweighted  pair group method with arithmetic mean (UPGMA) algorithms separated trypanozoon group from the T. congolense into two distinct clades. In conclusion,  this study suggests that the trypanozoon group of trypanosomes cause more canine trypanosomosis in the study area and suggests inclusion of dogs in strategic planning for control and eradication of trypanosomosis in sub-Sahara African countries. 
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用RRNA部分内转录间隔-1基因分析尼日利亚犬感染锥虫的分子鉴定
锥虫病正在成为包括尼日利亚在内的撒哈拉以南非洲地区犬类面临的主要健康挑战,但关于尼日利亚感染犬类的锥虫分子特征的报告很少。我们通过聚合酶链反应(PCR)和对锥虫核糖体RNA (ITS1 rRNA)内部转录间隔-1部分区域的序列分析,对自然感染犬中检测到的锥虫进行了鉴定。赠送给尼日利亚恩苏卡的尼日利亚大学兽医教学医院(UNVTH)进行检查和治疗的动物取样进行实验室检测。从显微镜下确认为锥虫感染阳性的21只狗的血液样本中提取DNA。对ITS-1进行PCR扩增和双向测序。16个样本条带较好,但其中一个条带序列不可读。NCBI上BLAST检索序列数据分析发现,从感染犬的分析样本中,刚果体、布氏冈比亚体和伊文氏体的感染率分别为4.8%、4.8%和91.4%。虽然前爆炸冲击t brucei组由于t . evansi和t . b . gambiense没有足够的歧视性权力区分亚种1。锥虫的排列序列多态性较小。采用UPGMA算法推导的系统发育树将锥虫类群与刚果锥虫类群划分为两个不同的支系。总之,本研究表明,在研究地区,锥虫的锥虫群导致更多的犬锥虫病,并建议将犬纳入撒哈拉以南非洲国家控制和根除锥虫病的战略规划中。
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