福建漳州流浪犬、猫芽囊原虫感染调查及基因分型

Investigation of Blastocystis spp. infection in stray dogs and cats and the parasite genotyping in Zhangzhou, Fujian

为了解福建省漳州市部分流浪动物收容中心犬、猫芽囊原虫感染情况,采集漳州地区救助犬、猫粪样并提取DNA,以芽囊原虫核糖体小亚基RNA (SSU rRNA)为引物进行PCR扩增。对阳性样品进行Sanger测序,根据GenBank公共数据库资源进行BLAST序列比对分析,采用MAGE 7.0软件构建系统进化树。共采集猫粪121份、犬粪119份。经PCR检验共7份样品阳性,其中犬芽囊原虫阳性率为5.04%(6/119),猫芽囊原虫阳性率为0.82%(1/121),犬、猫间阳性率差异具有统计学意义(χ2=4.153,P <0.05)。不同月龄的犬间、猫间,以及不同性别的犬间、猫间芽囊原虫阳性率差异均无统计学意义(χ2=2.149、 0.340、0.818、1.176,P> 0.05)。仅1份犬粪样的PCR扩增序列测序成功,为SSU rRNA基因序列YN-145株序列,通过BLAST序列比对,与人兽共患型芽囊原虫ST3亚型(GenBank登录号分别为MW404497.1、MW242639.1、MW767066.1)的一致性为95%~97%。系统进化树中YN-145序列与ST3形成同一聚类分支。

To investigate the prevalence of Blastocystis infections among dogs and cats at stray animal shelters in Zhangzhou City,Fujian Province,fecal samples were collected from rescued dogs and cats in Zhangzhou City,and DNA was extracted.Blastocystis ribosomal small subunit RNA(SSU rRNA)was used as a primer for PCR amplification.The positive samples were sent for Sanger sequencing,followed by BLAST sequence alignment analysis using the GenBank public database resources.A phylogenetic tree was constructed using MEGA 7.0 software.A total of 121 cat fecal samples and 119 dog fecal samples were collected.PCR test showed that the Blastocystis positivity rate of 5.04%(6/119)in dogs and 0.82%(1/121)in cats.The difference between dogs and cats was statistically significant(χ2=4.153,P<0.05).Only one fecal sample from a dog was successfully sequenced by PCR amplification of the SSU rRNA gene,yielding the YN-145 sequence.Sequence alignment using BLAST showed a high similarity of 95%-97%with zoonotic Blastocystis ST3 subtype sequences(GenBank accession numbers MW404497.1,MW242639.1,and MW767066.1).Phylogenetic analysis revealed that the YN-145 sequence formed a separate cluster within the ST3 subtype.