| Ticks as a kind of exclusively hematophagous ectoparasites, is the second important vector of disease transmitted in the world. Of which hard ticks are numerous.hazard and widespread distributing in China. The tick H aemaphysalis longicornis distributed both in the north and south China, however, Haemaphysalis asiaticum is mainly inhabiting the northwest China and Rhipicephalus haemaphysaloides is one of overwhelming superiority ticks of South China. Because of their roles in transmission of a wide range of human or animal pathogens, they had been intensively studied in control strategies, but it’s a shame that we almost knew nothing about the normal midgut microbiota of these ticks. Therefore, the present study was conducted to survey the midgut bacteria flora of three tick species, and also establishes foundation for further study the effect of intestinal bacteria on the tick physiology and pathogen transmission. Finally.these bacteria could be a potential way of reseach for new tick control methods and controlling tick-borne disease.In this study, the bacterial communities in adults gut of non-blood-fed lab-reared hard ticks, H. longicornis, H.asiaticum, R.haemaphysaloides were analysed using the traditional separation culture method and PLEX-ID technology. In addition,we also researched the diversity of the intestinal bacterial communities of blood-fed lab-reared and wild-caughted H. longicorni by16S rDNA sequencing combining with the system classification methods.The main results are as follows:1.A total of22strains were isolated from intestinal of three non-blood-fed Ixodids,of which7from H. longicornis,11from H. asiaticum,6from R. haemaphysaloides.These strains were classified into twelve different genera according to the characteristies of colonial morphology, mycelial, physiological-biochemical identification and16S rDNA phylogeny. They are Kocuria, Staphylococcus, Brevibacterium, Alcaligenes, Caryophanon, Microbacterium, Dielzia, Pseudomona, Corynebacterium, Moraxella, Streptomyces.2.Researching on the growth characteristics and pathogenic experiment of common strains Kocuria and Staphylococcus, which isolation from three hard ticks midgut, and found that both of them had no pathogenic for mice as well as the optimum growth temperature were about30℃. As for the most common genera it remains to clear whether a novel species of the genus Kocuria.3.The total of20strains were isolated from intestinal of blood-fed H. longicornis, of which13laboratory reared from H. longicornis,7from wild-caughted H, longicornis. And the common genus are bacillus, Staphylococcus and Corynebacterhum.4.PLEX-1D analysis the intestinal flora of three hard ticks and demonstrated that there are existed a lot of microorganism in tick’s midgut, especially in the intestinal of H. asiaticum. However.most of them were pathogenic bacteria.The results showed that there have large differences in using two different methods to analysis the midgut bacteria flora in three lab-reared hard ticks. Except for the common poit was numerous bacteria been found in H. asiaticum midgut, whereas relatively found little in the two other ticks. On the one hand it showed the rich microbiota diversity existed in the intestines of ticks. On the other hand,it also showed that each method has certain selectivity and limitations, There are a lot of microorganisms in the intestines of ticks can not be cultured and isolated by tridational method. PLEX-ID technology may also not be suitable for microbial identification in any areas. |
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