细口杯环线虫形态学和分子特征分析

doi:10.16431/j.cnki.1671-7236.2019.01.032

Abstract

Abstract:

In order to investigate the taxonomic status and phylogenetic relationship of Cylicocyclus leptostomum (C.leptostomum),this study used a digital microscope to observe the morphology of C. leptostomum,the ribosomal DNA internal transcribed spacer (rDNA-ITS) sequence was amplified by PCR,and 12 other ITS sequences of Strongylid nematodes were downloaded from GenBank,and the phylogenetic tree was constructed by maximum likelihood (ML) using Strongylus equinus as the outer group.The results showed that the nematode was medium-sized,the cyst was cylindrical,the width was greater than the depth,with small teeth at the bottom;The front end of the pouch wall was thin,and the base of the back end had obvious hoop-shaped thickening;The outer leaf crown was composed of 20 to 24 leaflets,and the inner leaf crown was composed of 50 to 60 leaflets;The esophagus funnel was smaller;Male reproductive cone was longer,conical;The female tail was straight,tail tip was finger-shaped;The total length of the measured ITS sequence was 837 bp,of which ITS1 was 366 bp,5.8S was 153 bp,and ITS2 was 318 bp.The GC content of ITS1 (46.0%) was significantly higher than that of ITS2 (39.8%);By BLAST homology alignment analysis,the ITS sequence of this study was 99.85% homologous with the same nematode sequence registered in GenBank (accession No.:AJ004849 and Y08587),and the homology with C.ashworthi,up to 99.0%,the homology with other nematodes in the genus was only 93.33% to 98.45%.The interspecies differences in ITS sequences were much larger than intraspecies differences.Phylogenetic analysis showed that C.leptostomum was closely related to C.ashworthi,and it was relatively distant from other nematodes in the genus.In summary,the ITS sequence could be used as a molecular genetic marker for identifying parasitic nematodes.It was confirmed that the specimen was a C.leptostomum,and the ITS sequence of C.leptostomum was reported for the first time in China,which would lay a foundation for further studies on this species.

Key words: Cylicocyclus leptostomum; morphological description; rDNA-ITS; phylogenetic analysis

CLC Number:

S852.73⁺1

HU Lili, SUN Yan, LIU Yingying, WANG Chen, BU Yanzhen. Morphological and Molecular Characteristics Analysis of Cylicocyclus leptostomum[J]. , 2019, 46(1): 271-278.

References

[1] 卜艳珍.河南省马圆线虫研究[M].郑州:河南科学技术出版社,2016. BU Y Z.Studies on Strongyloid Nematodes in Henan Province[M].Zhengzhou:Henan Science and Technology Press,2016.(in Chinese)
[2] 张路平,孔繁瑶.马属动物的寄生线虫[M].北京:中国农业出版社,2002. ZHANG L P,KONG F Y.Parasitc nematodes from Equus spp[M].Beijing:China Agriculture Press,2002.(in Chinese)
[3] LICHTENFELS J R,KHARCHENKO V A,DVOJONS G M.Illustrated identification keys to strongylid parasites (Strongylidae:Nematoda) of horses,zebras and asses (Equidae)[J].Veterinary Parasitology,2008,156(1-2):4-161.
[4] LICHTENFELS J R,KHARCHENKO V A,SOMMER C,et al.Key characters for the microscopical identification of Cylicocyclus nassatus and Cylicocyclus ashworthi (Nematoda:Cyathostominae) of the horse,Equus caballus[J].Journal of Helminthological Society of Washington,1997,64(1):120-127.
[5] MORARIU S,MEDERLE N,BADEA C,et al.The prevalence,abundance and distribution of cyathostomins (small stongyles) in horses from Western Romania[J].Veterinary Parasitology,2016,223:205-209.
[6] 沈杰,黄兵.中国家畜家禽寄生虫名录[M].北京:中国农业科学技术出版社,2004. SHEN J,HUANG B.A List of Parasites for Livestock and Poultry in China[M].Beijing:China Agricultural Science and Technology Press,2004.(in Chinese)
[7] ANJOS D H D,RODRIGUES M D L.Diversity of the infracommunities of strongylid nematodes in the ventral colon of Equus caballus from Rio de Janeiro state,Brazil[J].Veterinary Parasitology,2006,136(3-4):251-257.
[8] BU Y Z,NIU H X,ZHANG L P.Phylogenetic analysis of the genus Cylicocyclus (Nematoda:Strongylidae) based on nuclear ribosomal sequence data[J].Acta Parasitologica,2013,58(2):167-173.
[9] 段红,高俊峰,刘泽轩,等.四种三齿线虫核糖体ITS序列的比较分析及亲缘关系探讨[J].黑龙江八一农垦大学学报,2015,27(4):54-57. DUAN H,GAO J F,LIU Z X,et al.Comparative analysis and phylogenetic relationships among four species of genus Triodontophorus based on sequences of internal transcribed spacer of ribosomal DNA[J].Journal of Heilongjiang Bayi Agricultural University,2015,27(4):54-57.(in Chinese)
[10] SÁNDOR D,MOLNÁR K,GIBSON D I,et al.An investigation of the host-specificity of metacercariae of species of Apophallus (Digenea:Heterophyidae) in freshwater fishes using morphological,experimental and molecular methods[J].Parasitology Research,2017,116(11):3065-3076.
[11] 卜艳珍,勾利美,赵鹏飞,等.3种冠环线虫rDNA-ITS的PCR扩增及序列分析[J].中国畜牧兽医,2012,39(10):33-37. BU Y Z,GOU L M,ZHAO P F,et al.PCR amplification and sequence analysis of rDNA-ITS of three species of the genus Coronocyclus[J].China Animal Husbandry & Veterinary Medicine,2012,39(10):33-37.(in Chinese)
[12] REINEMEYER C R,PRADO J C,ANDERSEN U V,et al.Effects of daily pyrantel tartrate on strongylid population dynamics and performance parameters of young horses repeatedly infected with cyathostomins and Strongylus vulgaris[J].Veterinary Parasitology,2014,204(3-4):229-237.
[13] HINNEY B,WIRTHERLE N C,KYULE M,et al.Prevalence of helminths in horses in the state of Brandenburg,Germany[J].Parasitology Research,2011,108(5):1083-1091.
[14] HUNG G C,CHILTON N B,BEVERIDGE I,et al.A molecular systematic framework for equine strongyles based on ribosomal DNA sequence data[J].International Journal for Parasitology,2000,30(1):95-103.
[15] GASSER R B,WOODS W G,BJORN H.PCR-based SSCP to distinguish Oesophagostomum dentatum from O.quadrispinulatum developmental stages[J].International Journal for Parasitology,1998,28(12):1903-1909.
[16] 张路平,孔繁瑶.中国动物志线虫纲杆形目圆线亚目(二)[M].北京:科学出版社,2014. ZHANG L P,KONG F Y.Fauna Sinica Nematoda Rhabditida Strongylata (Ⅱ)[M].Beijing:Science Press,2014.(in Chinese)
[17] LICHTENFELS J R,KHARCHENKO V A,KRECEK R C,et al.An annotated checklist by genus and species of 93 species level names for 51 recognized species of small strongyles (Nematoda:Strongyloidea:Cyathostominea) of horses,asses and zebras of the world[J].Veterinary Parasitology,1998,79(1):65-79.
[18] 张淑熠,吴杰,郑刘梦,等.卡拉干斯齿线虫的形态及ITS和COⅠ序列分析[J].动物医学进展,2016,37(9):21-24. ZHANG S Y,WU J,ZHENG L M,et al.Morphological redescription and sequence analysis of ITS and COⅠ of Skrjabinodentus caragandicus[J].Progress inVeterinary Medicine,2016,37(9):21-24.(in Chinese)
[19] SERKHEL S P,GUPTA K S,KAUSHIK J,et al.Molecular characterization of internal transcribed spacer 1(ITS 1) region of different Trypanosoma evansi isolates of India[J].Journal of Parasitic Diseases,2017,41(2):527-533.
[20] ZHU X Q,D'AMELIO S,PALM H W.SSCP-based identification of members within the Pseudoterranova decipiens complex (Nematoda:Ascaridoidea:Anisakidae) using genetic markers in the internal transcribed spacers of ribosomal DNA[J].Parasitology,2002,124(6):615-623.
[21] 吕继洲,赵丽,赵昱林,等.拉合尔钝缘蜱的形态学及分子生物学鉴定[J].中国畜牧兽医,2017,44(4):1149-1158. LYU J Z,ZHAO L,ZHAO Y L,et al.Morphological and molecular biology identification of Ornithodoros lahorensis[J].China Animal Husbandry & Veterinary Medicine,2017,44(4):1149-1158.(in Chinese)
[22] WONGSAWAD C,WONGSAWAD P,SUKONTASON K,et al.Molecular phylogenetics of Centrocestus formosanus (Digenea:Heterophyidae) originated from freshwater fish from Chiang Mai province,Thailand[J].Korean Journal of Parasitology,2017,55(1):31-37.
[23] HUNG G C,GASSER R B,BEVERIDGE I,et al.Species-specific amplification by PCR of ribosomal DNA from some equine strongyles[J].Parasitology,1999,119(1):69-80.

Morphological and Molecular Characteristics Analysis of Cylicocyclus leptostomum

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	LI Zhongbo, HOU Qianghong, DUAN Deyong. Haplotypes Diversity,Genetic Differentiation and Phylogenetic Analysis of Aedes albopictus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1452-1460.
[2]	LIU Yutong, YAO Yanting, LI Ruiling, HAN Wenqi, SHI Ting, WANG Yidan, REN Yupeng. Analysis of Genetic Variation of Porcine Epidemic Diarrhea Virus in Parts of Southwest China from 2020 to 2021 [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2279-2290.
[3]	XU Minsheng, KE Haiyi, YANG Dongxia, SHI Keda, SUN Zeyi, NIU Jiawei, CHANG Xin, ZHAI Shaolun, ZANG Yingan, LI Chunling. Whole Genome Sequencing and Biological Characteristics Analysis of Multidrug-resistant Actinobacillus pleuropneumoniae GD2107 Strain [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(10): 4019-4031.
[4]	QI Yuzhuo, LIANG Lin, HAN Kun, LIANG Ruiying, JIA Yaxiong, CUI Shangjin. Genome Determination and Genetic Evolution Analysis of a Pigeon Circovirus Strain in Beijing [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(1): 32-42.
[5]	XU Ning, GE Guiyang, LI Dongli, LIU Yi, GONG Qinglong, MA Baoyi, SHENG Chenyan, SHI Kun, LI Jianming, LENG Xue, DU Rui. Isolation and Identification of Porcine Pseudorabies Virus and Genetic Variation Analysis of Partial Virulence Gene [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(5): 1794-1803.
[6]	TANG Ye, CHEN Yi, TIAN Xiaoyan, LYU Haifeng, GAN Junji. Molecular Characterization of 3'-terminal Genome of Canine Coronavirus Strains JS1706 and JS1712 [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(11): 4231-4241.
[7]	ZHANG Li, REN Weike, LU Chao, LI Xiuli, LI Fuqiang, TIAN Xiangxue, CHI Jingjing, WANG Lili, JIANG Shan, DONG Zhimin, YAN Minghua. Genetic Variation Analysis of gB, gC and gE Genes of Porcine Pseudorabies Viruses Isolated in Tianjin During 2015-2020 [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3741-3751.
[8]	LI Mingyang, FANG Qingli, HUANG Weijian. Molecular Epidemiology of Bovine Astrovirus in Buffalo from Guangxi [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(9): 2926-2934.
[9]	LI Jiajia, SONG Jianling. Identification and Complete Genomic Analysis of a Newcastle Disease Virus from a Laying Hen [J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(1): 221-228.
[10]	WU Kainian, WANG Li, LI Chenyang. Cloning and Bioinformatics Analysis of BCL2A Gene in Jinchuan Yak [J]. , 2018, 45(6): 1463-1470.
[11]	WANG Lili, ZHENG Li, LU Chao, REN Weike, ZHANG Li, LI Fuqiang, YAN Minghua. Phylogenetic Analysis of Porcine Circovirus Type 2 Isolated from Tianjin and Its Surrounding Areas [J]. , 2018, 45(5): 1331-1340.
[12]	GE Shikun, ZHANG Kaizhao, LU Rong, LIU Jianxin, NING Zhangyong. Genetic and Phylogenetic Analysis of Orf Virus Isolated in China from 2006 to 2016 [J]. , 2018, 45(2): 439-447.
[13]	XU Run, HUANG Huaxu, XU Xiaoming, LI Zhengtai, MA Lin, LAO Xiaoxiang, WU Xianshi. Cloning and Phylogenetic Analysis of VP2 Gene of Canine Parvovirus in Nanning Region [J]. , 2018, 45(12): 3346-3354.
[14]	ZHANG Hui-min, Muhamaitijiang·T, SHI Jun, WU Gui-ling, Gulizhati·Adili, JIA Er-ken, SU Zhan-qiang, RAN Duo-liang, Shayilan·Kayizha, LAN Ling, YAO Gang, LIU Jian-hua. Isolation and Identification, F Gene Cloning and Evolutionary Analysis of Newcastle Disease Virus Isolated from Wild Birds in Xinjiang [J]. , 2017, 44(9): 2755-2760.
[15]	LI Xiao-wen, LIU You-ming, LIANG Guo-zhi, MEI Min-min, HUANG Wen-jing, LI Wen-feng, HUANG Shu-jian, LI Zhi-li. Isolation, Identification of Four Newcastle Disease Virus Strains and Study on Their Biological Characteristics and Genetic Characteristics [J]. , 2017, 44(7): 1925-1933.