山羊源3种不同形态阴门盖捻转血矛线虫雌虫的多位点序列分析

doi:10.11843/j.issn.0366-6964.2022.09.028

Abstract

Abstract: To elucidate the genetic and evolutionary differences of Haemonchus contortus with morphological differences in the vulval flap, in this study, fresh worms were isolated from the abomasum of infected goats. The morphological characteristics of the vulval flap of 71 female worms were observed by microscope. Based on the ITS-1, ITS-2 and nad4 gene loci, PCR was used to amplify H. contortus with morphological differences in the vulval flap, and phylogenetic analysis was performed by comparing the sequences of multiple loci. Morphological observation showed there existed three morphologic types of vulval flap for female H. contortus isolated in this study, namely linguiform, spheroid-shaped and ligule-spheroid form, and statistical analysis showed that the proportion of the three types of the vulval flap was 45.07% (32/71), 50.70% (36/71) and 4.23% (3/71), respectively. The body length and the length from the vulva to the posterior of 43 females (20 linguiform, 20 spheroid-shaped and 3 ligule-spheroid form) were measured and analyzed. Statistical analysis showed that there was no significant difference among the body length of the H. contortus with different vulval flap (P>0.05), but a significant difference was found in the length from the vulva to the posterior of the nematode with spheroid-shaped vulval flap compared with the other two types (P<0.05). Sequence analysis of the ITS-1, ITS-2 and nad4 genes of H. contortus showed there existed different levels of base differences at the three gene loci among 9 samples with different vulval flaps, that there were 10, 13 and 43 polymorphic sites, and the nucleotide diversity was 0.85%, 1.34% and 1.97%, respectively. The phylogenetic analysis based on ITS-1 and ITS-2 suggested that the samples here and reference sequence of H. contortus in GenBank were grouped into the same evolutionary branch, indicating that the parasites isolated here belonged to H. contortus. The morphological difference on vulval flap of female H. contortus was observed here and these parasites, on ITS-1, ITS-2 and nad4 gene locus, shown different degrees of base mutation. The results of this study should provide reference data for species identification and molecular epidemiological studies of H. contortus.

Key words: Haemonchus contortus, vulval flap, morphology, genetic locus, sequence analysis

CLC Number:

S852.731

LIANG Gaoxing, RONG Shiqi, WANG Junwei, LI Yuan, YANG Xin, ZHAO Guanghui, SONG Junke. Multi-locus Gene Sequence Analysis of Female Haemonchus contortus with Morphological Differences in Vulval Flap from Goat[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(9): 3140-3148.

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Multi-locus Gene Sequence Analysis of Female Haemonchus contortus with Morphological Differences in Vulval Flap from Goat

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