Morphological characteristics and molecular identification of Heterakis sp. peacock origin

doi:10.12140/j.issn.1000-7423.2021.06.013

Abstract

Abstract:

Objective To analyze taxonomic characteristics of Heterakis sp. from peacocks collected from Hefei Wildlife Zoo using morphological and molecular identification methods. Methods The worms were isolated from the cecum of peacocks and were indentified according to the morphological characteristics. The genomic DNA of the worms was extracted, then 28S rRNA D2-D3 gene and second internal transcribed spacer (ITS2) were amplified by PCR, and sequenced, the homology were analyzed by BLAST tool in GenBank, the phylogenetic tree was constructed by using the neighbour-joining method. Results A total of 48 nematodes were isolated. Male worm was 5.34-7.38 mm in length, while female 7.30-10.28 mm, which was about 2-5 mm longer than male. The copulatory spines in male was 80.00-322.73 μm in length, the pubic valve in female located near the caudal end of the worm. Both males and females contain three lamellae on the head, with a short oral cavity and a esophagus with cylindrical anterior and expanded posterior esophageal bulb. The male contains two unequal copulatory spines at the caudal end, while the female has a thin and smooth caudal tip. The egg is oval-shaped and gray-brown in color, covered by two layers of egg shell, which is fairly thick and smooth-surfaced, containing a single embryonic cell inside. PCR results showed that the 28S rRNA D2-D3 and ITS-2 amplification products were 750 bp and 1 053 bp in length respectively. BLAST showed that the 28S rRNA D2-D3 sequence was 84.59% homologous to S. calotis (GenBank Acession No. LC133189.1, LC133188.1) and 84.46% to H. gallinae (GenBank Acession No. LC185992.1, LC185989.1, LC186015.1, LC186006.1); the ITS-2 sequences was 85.29% homologous to H. beramporia (GenBank Acession No. KU529974, LC592732, LC592731), 80.32% to H. gallinarum (GenBank Acession No. LC59279, LC5928003, LC592805, LC592797) and 79.85% to H. indica (GenBank Acession No. LC592808, LC592809, LC592809). Based on the 28S rRNA D2-D3 gene phylogenetic tree, the Heterakis isolated in this study was genetically closer to S. calotis and the H. gallinae, but not on the same branch; based on the ITS-2 gene phylogenetic tree, it was genetically closer to the H. beramporia, H. gallinarum and H. indica, but not on the same branch. Conclusion The Heterakis worm isolated from peacock is morphologically highly consistent with the reported finding, thus, is preliminarily cassified as the Heterakis sp., but the phylogenetic trees based on the marker 28S rRNA D2-D3 and ITS-2 present certain variance with the reported.

Key words: Heterakis, Morphological identification, Molecular identification, Phylogenetic tree, Peacock

CLC Number:

R383.19

LIU Lu-yao, YANG Cong-shan, ZHANG Xiang, CHENG Rong-rong, HONG Yu-hang, WU Cheng-long, SUN Meng, CHEN Mu-xin, AI Lin, XU Qian-ming. Morphological characteristics and molecular identification of Heterakis sp. peacock origin[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021, 39(6): 816-820.

Figures/Tables 6

Table 1

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特征Characteristics	雄虫 Male	雌虫 Female
体长/mm Body length/mm	5.34~7.38	7.30~10.28
最大体宽/μm Body width of maximum/μm	280.31~361.24	322.45~378.59
食道长度/μm Length of esophagus/μm	680.00~920.33	810.63~1 085.47
排泄孔到头端距离/μm Distance from drain hole to tip/μm	310.08~362.53	418.92~540.00
尾长/μm Caudal length/μm	254.30~350.12	703.78~56.22
交合刺长度/μm Copulatory spine length/μm	80.00~322.73
阴门到尾端距离/mm Distance from vulva to caudal end/mm		3.76~5.41