International Livestock Research Institute (ILRI), Nairobi, Kenya.
Washington State University, Pullman, WA, USA.
Parasit Vectors. 2022 May 7;15(1):161. doi: 10.1186/s13071-022-05279-7.
Among protozoan parasites in the genus Babesia, Babesia bigemina is endemic and widespread in the East African region while the status of the more pathogenic Babesia bovis remains unclear despite the presence of the tick vector, Rhipicephalus microplus, which transmits both species. Recent studies have confirmed the occurrence of R. microplus in coastal Kenya, and although B. bovis DNA has previously been detected in cattle blood in Kenya, no surveillance has been done to establish its prevalence. This study therefore investigated the occurrence of B. bovis in cattle in Kwale County, Kenya, where R. microplus is present in large numbers.
A species-specific multiplex TaqMan real-time PCR assay targeting two Babesia bovis genes, 18S ribosomal RNA and mitochondrially-encoded cytochrome b and B. bigemina cytochrome b gene was used to screen 506 cattle blood DNA samples collected from Kwale County for presence of Babesia parasite DNA. A sub-set of 29 B. bovis real-time PCR-positive samples were further amplified using a B. bovis-specific spherical body protein-4 (SBP-4) nested PCR and the resulting products sequenced to confirm the presence of B. bovis.
A total of 131 animals (25.8%) were found to have bovine babesiosis based on real-time PCR. Twenty-four SBP4 nucleotide sequences obtained matched to B. bovis with a similarity of 97-100%. Of 131 infected animals, 87 (17.2%) were positive for B. bovis while 70 (13.8%) had B. bigemina and 26 (5.1%) were observed to be co-infected with both Babesia species. A total of 61 animals (12.1%) were found to be infected with B. bovis parasites only, while 44 animals (8.7%) had B. bigemina only. Babesia bovis and B. bigemina infections were detected in the three Kwale sub-counties.
These findings reveal high prevalence of pathogenic B. bovis in a Kenyan area cutting across a busy transboundary livestock trade route with neighbouring Tanzania. The Babesia multiplex real-time PCR assay used in this study is specific and can detect and differentiate the two Babesia species and should be used for routine B. bovis surveillance to monitor the spread and establishment of the pathogen in other African countries where B. bigemina is endemic. Moreover, these findings highlight the threat of fatal babesiosis caused by B. bovis, whose endemic status is yet to be established. GRAPHICAL ABTRACT.
在巴贝虫属的原生动物寄生虫中,巴贝虫双芽型在东非地区流行且广泛存在,而更具致病性的巴贝虫牛型的状况仍不清楚,尽管存在传播这两种物种的壁虱媒介锐缘蜱。最近的研究证实了锐缘蜱在肯尼亚沿海地区的存在,尽管以前在肯尼亚牛的血液中检测到了巴贝虫牛型 DNA,但尚未进行监测以确定其流行率。因此,本研究调查了肯尼亚夸莱县牛中是否存在巴贝虫牛型,在该地区大量存在锐缘蜱。
使用针对两种巴贝虫牛型基因,18S 核糖体 RNA 和线粒体编码细胞色素 b 以及巴贝虫双芽型细胞色素 b 基因的种特异性多重 TaqMan 实时 PCR 检测法,筛选来自夸莱县的 506 份牛血 DNA 样本中是否存在巴贝虫寄生虫 DNA。从 29 份实时 PCR 阳性的巴贝虫牛型样本中选择了一个亚组,使用巴贝虫牛型特异性的球形蛋白 4(SBP-4)巢式 PCR 进一步扩增,然后对所得产物进行测序以确认存在巴贝虫牛型。
根据实时 PCR,共有 131 只动物(25.8%)被发现患有牛巴贝斯虫病。获得的 24 个 SBP4 核苷酸序列与巴贝虫牛型完全匹配,相似度为 97-100%。在 131 只感染动物中,87 只(17.2%)为巴贝虫牛型阳性,70 只(13.8%)为巴贝虫双芽型阳性,26 只(5.1%)同时感染了两种巴贝虫。共有 61 只动物(12.1%)仅感染了巴贝虫牛型,而 44 只动物(8.7%)仅感染了巴贝虫双芽型。在夸莱县的三个分区均检测到了巴贝虫牛型和巴贝虫双芽型感染。
这些发现揭示了肯尼亚一个地区存在高致病性巴贝虫牛型的流行率,该地区横跨与邻国坦桑尼亚的繁忙跨界牲畜贸易路线。本研究中使用的巴贝斯虫多重实时 PCR 检测法具有特异性,可检测和区分两种巴贝斯虫,应用于常规巴贝虫牛型监测,以监测该病原体在其他巴贝虫双芽型流行的非洲国家的传播和建立。此外,这些发现强调了由巴贝虫牛型引起的致命巴贝斯虫病的威胁,其流行状况尚未确定。