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基于18S rRNA基因序列对尼日利亚阿贝奥库塔自然感染犬体内检测到的罗氏巴贝斯虫的遗传多样性研究

Genetic diversity among Babesia rossi detected in naturally infected dogs in Abeokuta, Nigeria, based on 18S rRNA gene sequences.

作者信息

Takeet Michael I, Oyewusi Adeoye J, Abakpa Simon A V, Daramola Olukayode O, Peters Sunday O

出版信息

Acta Parasitol. 2017 Mar 1;62(1):192-198. doi: 10.1515/ap-2017-0023.

Abstract

Adequate knowledge of the genetic diversity among Babesia species infecting dogs is necessary for a better understanding of the epidemiology and control of canine babesiosis. Hence, this study determined the genetic diversity among the Babesia rossi detected in dogs presented for routine examination in Veterinary Hospitals in Abeokuta, Nigeria. Blood were randomly collected from 209 dogs. Field-stained thin smears were made and DNA extracted from the blood. Partial region of the 18S small subunit ribosomal RNA (rRNA) gene was amplified, sequenced and analysed. Babesia species was detected in 16 (7.7%) of the dogs by microscopy. Electrophoresed PCR products from 39 (18.66%) dogs revealed band size of 450 bp and 2 (0.95%) dogs had band size of 430 bp. The sequences obtained from 450 bp amplicon displayed homology of 99.74% (387/388) with partial sequences of 18S rRNA gene of Babesia rossi in the GeneBank. Of the two sequences that had 430 bp amplicon, one was identified as T. annulata and second as T. ovis. A significantly (p<0.05) higher prevalence of B. rossi was detected by PCR compared to microscopy. The mean PCV of Babesia infected dogs was significantly (p<0.05) lower than non-infected dogs. Phylogenetic analysis revealed minimal diversity among B. rossi with the exception of one sequence that was greatly divergent from the others. This study suggests that more than one genotype of B. rossi may be in circulation among the dog population in the study area and this may have potential implication on clinical outcome of canine babesiosis.

摘要

充分了解感染犬类的巴贝斯虫物种之间的遗传多样性,对于更好地理解犬巴贝斯虫病的流行病学和控制方法至关重要。因此,本研究确定了在尼日利亚阿贝奥库塔兽医医院接受常规检查的犬只中检测到的罗氏巴贝斯虫的遗传多样性。从209只犬中随机采集血液。制作现场染色薄涂片,并从血液中提取DNA。对18S小亚基核糖体RNA(rRNA)基因的部分区域进行扩增、测序和分析。通过显微镜检查在16只(7.7%)犬中检测到巴贝斯虫物种。39只(18.66%)犬的电泳PCR产物显示条带大小为450 bp,2只(0.95%)犬的条带大小为430 bp。从450 bp扩增子获得的序列与基因库中罗氏巴贝斯虫18S rRNA基因的部分序列显示出99.74%(387/388)的同源性。在两个具有430 bp扩增子的序列中,一个被鉴定为环形泰勒虫,另一个为绵羊泰勒虫。与显微镜检查相比,PCR检测到的罗氏巴贝斯虫患病率显著更高(p<0.05)。感染巴贝斯虫的犬的平均红细胞压积显著低于未感染犬(p<0.05)。系统发育分析显示,除了一个与其他序列有很大差异的序列外,罗氏巴贝斯虫之间的多样性极小。本研究表明,研究区域的犬群中可能存在不止一种罗氏巴贝斯虫基因型,这可能对犬巴贝斯虫病的临床结果有潜在影响。

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