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Glutathione S-transferases (GSTs) are phase II detoxification enzymes, which function via combining with pesticidal molecules and catalyzing the conjugation of molecules by thiol of glutathione, so as to protect tissues from oxidative stress damage. In the tick Haemaphysalis longicornis, glutathione S-transferases (HlGST and HlGST2) have been previously identified. However, the relationship between the transcription of glutathione S-transferases and the essential oil treatment in ticks remains unexplored. Hence, in the present study, the transcription profiles of HlGST and HlGST2 mRNAs were evaluated in H. longicornis after exposure to Cymbopogon citratus essential oil. At 24 h post-exposure of H. longicornis to different sublethal concentrations of C. citratus essential oil, there was significant difference (P = 0.0001) in the transcription of HlGST. Tukey’s test showed that HlGST was significantly induced after treatment with 1% C. citratus essential oil (P = 0.0002); whereas no significant difference (P = 0.3551) was detected after treated by 2% C. citratus essential oil. No significant difference (P = 0.4555) in the transcription of HlGST2 between the treatment and the control group of 50% ethanol. Nevertheless, the down-regulation of HlGST2 in the treatment groups versus the untreated control group was not significant (P = 0.1208). The above results imply that the essential oil can modulate the expression of GST mRNA, therefore, further understanding of the underlying mechanisms of the GST at the molecular level could contribute to the development of effective control measures for ticks and tick-borne diseases.

Background Haemaphysalislongicornis is an obligate hematophagous ectoparasite that transmits a variety of pathogens causing life-threatening diseases in humans and animals. Paramyosin (Pmy) is not only an invertebrate-specific myofibrillar protein but also an important immunomodulatory protein. Therefore, it is one of the ideal candidate antigens for vaccines. Methods We conducted two vaccine trials to evaluate the protective efficacy of Pmy recombinant protein (rPmy) and peptide vaccine (KLH-LEE). Each rabbit was immunized with three doses of rPmy or KLH-LEE adjuvanted with Freund’s complete/incomplete at 500 μg/dose at 2-week intervals before challenge with 40 female H.longicornis/rabbit. PBS plus adjuvant, Trx or KLH was used as control group. The antibodies of rabbits were detected by ELISA. Then, female ticks were fed on the rabbits until detachment. Results ELISA results showed that both vaccines induced rabbits to produce antibodies. Compared with the Trx group, the engorgement weight, oviposition and hatchability of the rPmy group decreased by 8.87%, 26.83% and 38.86%, respectively. On the other hand, engorgement weight, oviposition and hatchability of female ticks in the KLH-LEE group correspondingly resulted in 27.03%, 53.15% and 38.40% reduction compared with that of the KLH group. Considering the cumulative effect of vaccination on the evaluated parameters, results showed 60.37% efficacy of the rPmy vaccine formulation and 70.86% efficacy in the KLH-LEE group. Conclusions Pmy and particularly epitope LEE have potential for further development of an effective candidate vaccine to protect the host against tick infection. Graphical Abstarct

Haemaphysalis longicornis is distributed worldwide and transmits a variety of pathogens, causing human and animal disease. Use of chemical acaricides, as a primary tick control method, has several disadvantages, including acaricide resistance, environmental damage and residue accumulation in livestock. Development of a livestock vaccination aimed at a tick protective antigen could be an effective, labor-saving and environmentally-friendly method of reducing tick infestation and transmission of tick-borne pathogens. Lipocalins are low molecular weight proteins that play important roles in blood feeding, immune response and reproduction in ticks. In our study, the open reading frame (ORF) of a lipocalin homologue from H. longicornis (HlLIP) was successfully cloned, which consisted of 387 bp encoding a protein of 128 amino acids. The HlLIP protein sequence showed a close sequence homology with Ixodes persulcatus lipocalin. The HlLIP gene was constitutively detected in all developmental stages and in all tissues of the unfed female tick. The ORF of the HlLIP gene was sub-cloned into pET-32a (+) to obtain the recombinant protein (rHlLIP) and its immunogenicity was comfirmed by western blot. A vaccination trial on rabbits against H. longicornis infestation demonstrated that the rHlLIP protein could significantly prolong the period of tick blood feeding, and reduce tick engorged weight, oviposition and egg hatching rate. The vaccination efficacy of the rHlLIP protein was 60.17 % based on engorged weight, oviposition and egg hatching rate of ticks. The results obtained in this study demonstrate that rHlLIP protein is a promising antigen that could potentially be developed as a vaccine against H. longicornis infestation.