Identification, artificial rearing and observation of life cycle of Ixodes persulcatus in Inner Mongolia

doi:10.12140/j.issn.1000-7423.2024.03.010

Abstract

Abstract:

Objective To identify the morphological and molecular characteristics of Ixodes persulcatus collected from Inner Mongolia and to understand their developmental life cycle and biological characteristics through artificial rearing. Methods Ticks were collected from the northeastern region of Inner Mongolia and identified morphologically using a 3D super depth microscope system. Tick DNA was extract, and mitochondrial 12S rDNA and 16S rDNA sequences was amplifed by PCR. After sequencing the positive amplification products, BLAST alignment was performed on the sequences, and the tick mitochondrial 12S rDNA and 16S rDNA genes phylogenetic trees were constructed using the neighbor joining algorithm. Kunming mice were used as blood source to artificially rear the ticks at a temperature of (25 ± 3) ℃ and a relative humidity of 70%-90%. The biological characteristics of the tick life cycle at different stages were observed. In the egg hatching experiment, 30 eggs from each of 10 fully blood-fed female adult ticks were randomly selected, their hatching was observed, and the hatching rate was calculated. A total of 200 engorged larval ticks were randomly selected for the molting experiment, and their molting status was recorded and the molting rate was calculated. 100 engorged nymphal ticks were randomly selected for the nymphal ticks molting experiment and divided into 10 groups, with 10 ticks in each group. The molting situation was observed, and the molting rate was calculated. The comprehensive development rate of each stage after calculating the success rate of development was calculated. Results The morphological identification results showed that both the collected female and male ticks conform to the morphology of Ixodes. The PCR amplification and sequencing results showed that the 12S rDNA sequence with a length of 320 bp and the 16S rDNA sequence with a length of 455 bp were amplified from tick DNA. BLAST sequence alignment analysis showed that the amplified mitochondrial 12S rDNA sequences of female and male ticks had the highest homology with the sequences of I. persulcatus (GenBank: MF095801.1 and JF758624.1), with 99.69% and 99.09%, respectively. BLAST sequence alignment analysis showed that the amplified mitochondrial 16S rDNA sequences of female and male ticks had the highest homology with the sequences of I. persulcatus (GenBank: MH790201.1 and MH790200.1), with 99.75% and 99.50%, respectively. The genetic evolution analysis results showed that both female and male ticks clustered on the same branch as the sequences of the I. persulcatus. The observed life cycle of artificially reared I. persulcatus showed that the oviposition period of engorged female adult ticks is 12-17 days, with an average oviposition period of 14.6 days. The total number of eggs laid is about 1 510-1 970 per tick, with an average oviposition of about 1 817 per tick, and a daily average oviposition of about 124 per tick. The eggs hatched into larval ticks after 21-28 days, with an average hatching period of 24.8 days and a hatching rate of 89.7% (269/300). The blood-feeding period of the larval ticks is 3-5 days, with an average blood-feeding period of 4.5 days. After 18-25 days of molting, engorged larval ticks molt as nymphal ticks, with an average molting period of 22.8 days and a molting rate of 86.5% (173/200). The blood-feeding period of nymphal ticks is 5-8 days, with an average blood-feeding period of 6.3 days. Engorged nymphal ticks molted after 120-170 days to become adult ticks, the average molting period is 157.2 days, and the molting rate is 92.0% (92/100). It took an average of 241.6 days to develop to the next generation of adult ticks from the engorged female adult ticks laying eggs, with a comprehensive development rate of 71.4%. Conclusion The ticks collected from Inner Mongolia is confirmed to be I. persulcatus through morphological and molecular biology identification. Artificial feeding experiments were conducted to obtain the biological characteristics of the life cycle of the I. persulcatus, including eggs, larval ticks, nymphal ticks, and adult ticks.

Key words: Ixodes persulcatus, Morphological identification, Molecular biological identification, Artificial feeding, Life cycle

CLC Number:

R384.4

SUN Lianyang, CUI Hao, DONG Xiaonan, KANG Jiamei, DING Yulin, XI Juan, YANG Yang, HE Zhixiong, LIU Yonghong, ZHAO Li. Identification, artificial rearing and observation of life cycle of Ixodes persulcatus in Inner Mongolia[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2024, 42(3): 345-353.

Figures/Tables 6

Table 1

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基因 Gene	引物序列 Primer seqence	扩增长度/bp Amplification length/bp
12S rDNA	F：AAACTAGGATTAGATACCCT	320
	R：AATGAGAGCGACGGGCGATGT
16S rDNA	F：TTAAATTGCTGTRGTATT	455
	R：CCGGTCTGAACTCASAWC