| (一) Objective1.To research and establish comprehensive multi-disease surveillance mechanism of tick and host animals-based infected with pathogen of relevant tick-borne infectious disease.2.To describe the distribution pattern of major host animal of tick-borne infectious disease in Zhejiang province.3.To describe the epidemiology of infection of tick-borne disease of the rodents.4.To describe the epidemiology of infection of tick-borne disease in Zhejiang province.5.To analyze the epidemiology of distribution of tick-borne disease in different seasons.6.To analyze the geographic distribution of major tick-borne infectious disease.(二) Content and Method1.Conduct the field survey on types of major tick-borne host of vector tick in different geographic areas.2.Sample collected on site for classification and verification, liver and spleen sample of the rodents are collected and grinded for DNA extraction. Samples of Vector tick are grinded for DNA extraction; adopt methodology of molecular biology to test infection in Zhejiang province.3.Positive samples with clearer Electrophoresis bands selected for sequencing for each kind of disease in each area. Identify the disease category of positive sample after online comparison.4.Construct phylogenetic tree with MEGA5.0based on sequencing for systematic analysis. 5.Analysis of the pathogen infection in hosts and ticks in various seasons and areas with SAS9.0.(三) Results1.Host animal surveillance:We have deployed squirrel cage at six surveillance sites in four quarters for6755cage times and captured630rodents. Capture rate is9.33%.1st quarter118;2nd quarter148;3rd quarter;4th quarter127. It shows that Podiums agrarius and Niviventer confucianus are the dominant species outdoor, capture295Apodemus agrarius, which accounts for46.83%of the total,179Niviventer confucianus, which accounts for28.41%.2.Vector surveillance:Haemaphysalis have various parasitic hosts, including cow, sheep, dog and rat; Boophilus microplus has sole parasitic host, which only is captured in cow. The types of parasitic tick to the rodents are Haemaphysalis, Ixodes sinensis, lxodes granulatus Supino; domestic sheep has various of parasitic tick, including Haemaphysalis, Rhipicephalus haemaphysaloides tick, Ixodes sinensis, Ixodes granulatus Supino. Also, sole parasitic host to dog, including Rhipicephalus haemaphysaloides tick, Rhipicephalus Sanguineu, Haemaphysalis longicornis and Boophilus microplus are parasitic to the cow. Due to few tick number captured at monitoring site, so we didn’t conduct free vector surveillance all year round. In the first quarter, free tick surveillance conducted at six monitoring sites with the white cloth method, the density of free tick in the province is8piece/hour, the density of Xianju tops of all, which is32piece/hour, Haining, Longyou and Jiande is lowest, no free tick is captured yet.3.Laboratory testing:The comprehensive surveillance for study of the tick-borne infectious disease adopted the methodology of molecular biology to test each sample for six pathogens, including Bartonella, Babesia sp, Haemaphysalis longicornis, E. chaffeensis, Lyme spirochetes and rickettsia. All of which has been detected from the rodents. Babesia sp, Haemaphysalis longicornis, E. chaffeensis and rickettsia are only detected from tick.From the pathogen carried by the rat species, all the6pathogens are detected from Rattus confucianus from the rodents captured at each surveillance sites within the province. Whereas, three pathogens are found from Apodemus agrarius, Rattus Edwards and Microtus fortis respectively; one pathogen is found from Rattus rattoides and sewer rat. For the tick species with positive result, four pathogens are found in Haemaphysalis longicornis,l pathogen from Rhipicephalus Sanguineus.28tested for Bartonella positive from630samples of rodents. The positive rate is4.44%, The highest positive rate of Rattus confucianus is5.59%(10samples).22Babesia sp positive, the rate is3.49%. the highest rate is7.82%from Rattus confucianus (14samples);26Haemaphysalis longicornis positive, the rate is4.13%, The highest positive rate of Rattus confucianus is11.17%(20samples);8E. chaffeensis positive, the rate is1.27%. the highest positive rate of Rattus Edwards is11.76%(2samples);5Lyme spirochetes positive, rate is0.79%, Rattus confucianus tested for positive, rate is2.79%(5samples);4is rickettsia positive, the rate is0.63%, the highest positive rate is Rattus Edwards, which is up to5.88%(1sample). No Bartonella and Lyme positive has not been detected from779tick samples.6Babesia sp.positive found in Haemaphysalis longicornis. Total positive rate is0.77%,sub-tick positive rate is1.58%.12is Haemaphysalis longicornis positive, rate is1.54%, positive results were found in Ixodes sinensis and Haemaphysalis longicornis. The rate is8.99%(8samples);8E. chaffeensis positive, rate is1.03%; positive result found in Haemaphysalis longicornis, rate is2.11%(8samples);7samples are positive, rate is0.9%, positive results were found in Haemaphysalis longicornis, Ixodes sinensis and Rhipicephalus Sanguineus, rate is0.79%(3samples)、3.37%(3samples)'1.03(1sample)4.Do the sequencing for all the samples tested, and then analyze evolution on sequence data. The tested Haemaphysalis longicornis and E. chaffeensis are distributed comparatively scatted in the evolutionary trees. Whereas, partial sequencing is genetically related with neighboring countries, Yunan and U.S. Bartonella and Bartonella Henselae, is genetically related. It is found that pathogen species of micro-Babesia sp. and Phosphorus, via sequencing comparison, exist in the province. Borrelia burgdorferi sensu lato is genetically related with the sequencing tested in the province.5.Comparison of Positive samples tested among different kinds of rodents Perform Fisher exact statistical analysis for all the pathogens tested in different types of rodents, Babesia sp. positive rate in Rattus has significance with other rats species.(P<0.05), HGA positive rate in Rattus has significance with other rats species(P<0.05), HME positive rate in Rattus Edwards has significance with other rates species (P<0.05).6.Comparing positive results at different surveillance sites, Bartonella positive rate in Haining has significance with other surveillance sites. HGA positive rate in Jiande has significance with other surveillance sites. HME positive rate in Xianju has significance with other surveillance sites.7. Comparison of Positive results in different season, Samples with positive result tested in different season has been analyzed statistically, it shows that positive rate of bartonella and rickettsia has no significance, positive rate of Babesia sp.in the first quarter has significance compared with other seasons (P<0.05), HGA positive rate in the2nd and4th quarter has significance with other seasons (X2=15.261, P<0.05), HME positive rate in the first quarter has significance with other seasons (P<0.05), positive rate of Lyme disease in the first quarter has significance with other seasons (P<0.05)(四) Conclusion1.Implementation of tick-borne infectious disease surveillance system makes surveillance activities more standard and integrative. Furthermore, all the samples have been taken fully utilized, multi-pathogen test has been performs for each sample, including Bartonella, Babesia sp., Haemaphysalis longicornis, E. chaffeensis Lyme disease and rickettsia.2.Pathogen of Bartonella, Babesia sp., Haemaphysalis longicornis, E. chaffeensis Lyme disease and rickettsia exist in Zhejiang province. Furthermore, there is certain infection prevalence among rodents and tick species, it is possible that natural foci exists in Zhejiang province.3.Rattus is dominant species outdoor in Zhejiang province, the infection spectrum and rate of tick-borne infectious disease ranks first among the rodents captured in this study. Other rats species such as Rattus Edwards and Apodemus agrarius are infected also.4.Compare positive rate of those infected with tick-borne diseases in different areas, it is found that high positive rate of Bartonella happened in Haining, which locates at northern plain area in Zhejiang province. High HGA positive rate is found in Jiande, HME positive rate is high in Xianju, has significance with other areas. It indicates that there are various types of tick-borne infectious disease may exist in different geographical area in the province.5.Compare different tick-borne infectious disease in different seasons, it is found that positive rate of Bartonella and rickettsia has no difference in seasonality, positive rate of Babesia sp.in the first quarter has significance compared with other seasons. HGA positive rate in2nd and4th quarter has significance with other seasons. HME positive rate in the first quarter has significance with other seasons. Positive rate of Lyme disease in the first quarter has significance with other seasons. It demonstrates that tick-borne infection host animals and infection medium most happened in the first quarter in the province, there might some detected in other seasons, but with lower positive rate.6.1t is necessary to conduct health communication among high risk population such as working staff in forestry area to prevent the disease. |
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