Abstract
Ticks are blood-sucking ectoparasites of great medical and veterinary significance that can transmit bacteria, protozoa, fungi and viruses, causing a variety of human and animal diseases worldwide. In the present study, the intestinal bacterial flora associated with Haemaphysalis flava ticks in different developmental stages were analyzed using polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE). Eleven distinct DGGE bands were found using PCR-DGGE method. Sequences analyses indicated that they belonged to Bacillus cereus, Candidatus rickettsia, Erwinia sp., Klebsiella pneumoniae, Pectobacterium carotovorum, Pseudomonas aeruginosa, Rickettsia peacockii, Rickettsia helvetica, Rickettsia slovaca, Staphylococcus simulans and Uncultured bacterium clone. Our find that the K. pneumoniae and P. aeruginosa isolates were presented in all H. flava ticks in different developmental stages. The present results indicated that zoonotic pathogens are present in H. flava ticks in Henan province, China. To our knowledge, this is the first report on intestinal bacterial flora associated with H. flava ticks in China.
Competing interests
The authors declare that they have no competing interests.
Ethic statement
All animals were handled in strict accordance with the Good Animal Practice requirements of the Animal Ethics Procedures and Guidelines of the People’s Republic of China.
Acknowledgments
This research was financially supported by grant from the National Natural Science Foundation of China (No. 31372431).
References
Dantas-Torres F., Chomel B.B., Otranto D. 2012. Ticks and tickborne diseases: a one health perspective. Trends Parasitology, 28, 437–446. 10.1016/j.pt.2012.07.003Search in Google Scholar
Diuk-Wasser M.A., Vannier E., Krause P.J. 2016. Coinfection by Ixodes Tick-Borne Pathogens: Ecological, Epidemiological, and Clinical Consequences. Trends Parasitology, 32, 30–42. 10.1016/j.pt.2015.09.008Search in Google Scholar
Fang L.Q., Liu K., Li X.L., Liang S., Yang Y., Yao H.W., et al. 2015. Emerging tick-borne infections in mainland China: an increasing public health threat. Lancet Infectious Diseases, 15, 1467–1479. 10.1016/S1473-3099(15)00177-2Search in Google Scholar
Gao X., Nasci R., Liang G. 2010. The neglected arboviral infections in mainland China. PLoS Neglect Tropical Diseases, 4, e624. 10.1371/journal.pntd.0000624Search in Google Scholar
Karbowiak G., Biernat B., Werszko J., Rychlik L. 2016. The transstadial persistence of tick-borne encephalitis virus in Dermacentor reticulatus ticks in natural conditions. Acta Parasitologica, 61, 201–203. 10.1515/ap-2016-0028Search in Google Scholar
Kušar D., Avguštin G. 2012. Optimization of the DGGE band identification method. Folia Microbiology, 57, 301–306. 10.1007/s12223-012-0130-9Search in Google Scholar
Liu L., Li L., Liu J., Hu Y., Liu Z., Guo L., Liu J. 2013. Coinfection of Dermacentor silvarum olenev (acari: ixodidae) by Coxiella-Like, Arsenophonus-like, and Rickettsia-like symbionts. Applied and Environment Microbiology, 79, 2450–2454. 10.1128/AEM.03575-12Search in Google Scholar
Liu Q., He B., Huang S.Y., Wei F., Zhu X.Q. 2014. Severe fever with thrombocytopenia syndrome, an emerging tick-borne zoonosis. Lancet Infectious Diseases, 14, 763–772. 10.1016/S1473-3099(14)70718-2Search in Google Scholar
Moreno C.X., Moy F., Daniels T.J., Godfrey H.P., Cabello F.C. 2006. Molecular analysis of microbial communities identified in different developmental stages of Ixodes scapularis ticks from Westchester and Dutchess Counties, New York. Environment Microbiology, 8, 761–77210.1111/j.1462-2920.2005.00955.xSearch in Google Scholar PubMed
Opalińska P., Wierzbicka A., Asman M. 2016. The PCR and nested PCR detection of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Babesia microti in Dermacentor reticulatus F. collected in a new location in Poland (Trzciel, Western Poland). Acta Parasitologica, 61, 849–854. 10. 1515/ap-2016-0117Search in Google Scholar
Schabereiter G.C., Lubitz W., Rölleke S. 2003. Application of broad-range 16S rRNA PCR amplification and DGGE fingerprinting for detection of tick-infecting bacteria. Journal of Microbiology Methods, 52, 251–26010.1016/S0167-7012(02)00186-0Search in Google Scholar
Sun J., Lin J., Gong Z., Chang Y., Ye X., Gu S., et al.2015. Detection of spotted fever group Rickettsiae in ticks from Zhejiang Province, China. Experimental and Applied Acarology, 65, 403–411. 10.1007/s10493-015-9880-9Search in Google Scholar PubMed PubMed Central
Tveten A.K., Riborg A., Vadseth H.T. 2013. DGGE identification of microorganisms associated with Borrelia burgdorferi Sensu Lato-or Anaplasma phagocytophilum-infected Ixodes ricinus ticks from northwest Norway. International Journal of Microbiology, 2013, 805456. 10.1155/2013/805456Search in Google Scholar PubMed PubMed Central
Tveten A.K., Sjåstad K.K. 2011. Identification of bacteria infecting Ixodes ricinus ticks by 16S rDNA amplification and denaturing gradient gel electrophoresis. Vector-Borne and Zoonotic Diseases, 11, 1329–1334. 10.1089/vbz.2011.0657Search in Google Scholar PubMed
Van Overbeek L., Gassner F., Van Der Plas C.L., Kastelein P., Nunesda Rocha U., Takken W. 2008. Diversity of Ixodes ricinus tick-associated bacterial communities from different forests. FEMS Microbiology Ecology, 66, 72–84. 10.1111/j. 1574-6941Search in Google Scholar
Wu X.B., Na R.H., Wei S.S., Zhu J.S., Peng H.J. 2014. Distribution of tick-borne diseases in China. Parasties & Vectors, 6, 119. 10.1186/1756-3305-6-119Search in Google Scholar PubMed PubMed Central
Xu X.L., Cheng T.Y., Yang H., Yan F. 2015. Identification of intestinal bacterial flora in Rhipicephalus microplus ticks by conventional methods and PCR-DGGE analysis. Experimental and Applied Acarology, 66, 257–268. 10.1007/s10493-015-9896-1Search in Google Scholar PubMed
Xu X.L., Cheng T.Y., Yang H., Liao Z.H. 2016. De novo assembly and analysis of midgut transcriptome of Haemaphysalis flava and identification of genes involved in blood digestion, feeding and defending from pathogens. Infectious Genetics and Evolution, 38, 62–72. 10.1016/j.meegid.2015.12.005Search in Google Scholar PubMed
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