Survey On Ticks And Detection Of New Bunyavirus In The Endemic Areas Of Fever, Thrombocytopenia And Leukopenia Syndrome In Henan Province

Objective To survey the distribution, speceies of ticks and detect New bunyavirus in the endemic areas of fever, thrombocytopenia and leukopenia syndrome (FTLS) in Henan province.Methods The free ticks were collected manually with white cloth (100cm*100cm) from the grassland and bush, and the parasitic ticks were collected from the host skin of domestic animals by hand searching. All the collected ticks were collected into the same tube by the habitat of each month, then they were identification morphologically in the laboratory and counted the density of the different habitat of each month (only/artifical banner hours or only/each in animals). The New bunyavirus were isolated from the ticks affter grindded on the ice. at the same time. followed by detection of New bunyavirus with PCR. After the PCR products were purified and cloned, the determination nucleotides sequences were assisted proofreading with the SeqMan of DNAStar software. MegAlign software was used to analyzed the homology. The nucleotides sequences were compared and analyzed with the sequences in GenBank using DNAStar7.0software.Results12388ticks were collected in the two survey areas, including7172free ticks and5216parasitic ticks, belonged to2families,5geniuses and6species. The collected ticks were identification morphologically as Haemaphysalis longicornis, H.detritum, H.anatolicum anatolicum, Boophilus microplus, Rhipicephalus sanguineus and Ornithodoros lahorensis. The relative dominance of Haemaphysalis longicornis, H.detritum, H.anatolicum anatolicum, Boophilus microplus and Rhipicephalus sanguineus were65.94%(8169/12388),1.11%(138/12388),1.32%(164/12388),5.13%(635/12388), and0.44%(54/12388) respectively.7172free ticks were collected with white cloth (100cm*100cm) from the grassland and bush in86artificial/hours, and the average density was83.40(labor/hour). The free ticks were active from March to October and in July the density of up was651(manual/h).5216parasitic ticks were collected from the host skin of dornestic animals by hand searching, including36ticks were found in the44buffalo, the average tick density was0.8/buffalo,827ticks were found in the39cattle, the average tick density was21/cattle,3506ticks were found in the163sheep, the average tick density was21.51/sheep, and203ticks were found in the39dogs, the average tick density was5.21/dog. The parasitic ticks were active from March to December and in June the highest density was27.15(only/domestic animals).77groups of tick (825ticks) were cultured and isolated in vero cell. At the same time, they were detected by PCR. None new bunyavirus was isolated and11groups were positive by PCR. After11positive PCR products were sequenced with purified and cloned, the amplified gene fragment was600bps, and nucleic acid similarity of11specimens were93%-100%and there’s also a high similarity (100%) between the samples and new bunyavirus isolates (Accession No. HQ419214.1), followed by99%(Accession No.JF682778.1), and a minimum of93%(Accession No. HQ179742.1). The sequences amplified from the ticks was confirmed that was the new bunyavirus.Conclusion1The ticks in the endemic areas of fever, thrombocytopenia and leukopenia syndrome (FTLS) in Henan province belonged to2families,5geniuses and6species. There were five kinds belonged to Ixodinae, including Haemaphysalis longicornis, H.detritum, H.anatolicum anatolicum, Boophilus microplus and Rhipicephalus sanguineus. The Ornithodoros lahorensis belonged to Argasidae.2Two new records of ticks were found in Henan province, which were Ornithodoros lahorensis and H.anatolicum anatolicum. and the Haemaphysalis longicornis was found as the predominant species.3The free ticks exist in the outside environment from March to October, and the peak appears from May to August, which disappeared after October. The parasitic ticks can parasitic in the host skin of domestic animals throughout the year, and the peak appears from May to September.4The nucleic acid detection of tick was positive by PCR, which means that ticks may be the vector of New bunyavirus.