• Users Online: 338
  • Home
  • Print this page
  • Email this page
Home About us Editorial board Ahead of print Current issue Search Archives Submit article Instructions Subscribe Contacts Login 

Table of Contents
Year : 2021  |  Volume : 58  |  Issue : 3  |  Page : 183-192

Current status and future prospects of multi-antigen tick vaccine

1 Entomology Laboratory, Parasitology Division, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, India
2 Department of Veterinary Parasitology, College of Veterinary Science & Animal Husbandry, Junagadh Agricultural University, Junagadh, Gujarat, India

Date of Submission05-Mar-2020
Date of Decision28-Dec-2020
Date of Web Publication15-Feb-2022

Correspondence Address:
Srikant Ghosh
Entomology Laboratory, Parasitology Division, ICAR-Indian Veterinary Research Institute, Izatnagar-243122, Bareilly
Login to access the Email id

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0972-9062.321739

Rights and Permissions

Ticks are blood sucking ectoparasite that transmit several pathogens to humans and animals. Tick management focusing on use of chemicals has several drawbacks including development of multi-acaricide resistant tick populations. To minimize the use of chemicals on animals and on the environment, immunization of natural hosts is considered a viable component of Integrated Tick Management System. Most of the tick vaccine trials are focused on single antigen immunization directed against homologous challenge. From commercial point of view, vaccination against one given tick species is not a feasible option. In this context, multi-antigen vaccines comprising of candidate antigens of multiple tick species or both ticks and tick-borne pathogens have commercial potential. Different strategies are considered for the development of multi-antigen tick and/or tick-borne pathogen vaccines. Further, the efficacy of vaccine can be improved by adopting the ‘omics’ tools and techniques in selection of novel antigens and efficient delivery like Lipid Nano Particle (LNP)-mRNA vaccines, viral vector vaccine, live vector vaccine etc. into the host. The subject has been reviewed to address the current status of multi antigen tick vaccines and formulations of the future strategies for the control of TTBDs of human and animals.

Keywords: Tick; tick-borne pathogen; multi-antigen tick vaccine; chimeric vaccine; co-vaccination; multi-antigen oral vaccines.

How to cite this article:
Parthasarathi B C, Kumar B, Ghosh S. Current status and future prospects of multi-antigen tick vaccine. J Vector Borne Dis 2021;58:183-92

How to cite this URL:
Parthasarathi B C, Kumar B, Ghosh S. Current status and future prospects of multi-antigen tick vaccine. J Vector Borne Dis [serial online] 2021 [cited 2023 Mar 29];58:183-92. Available from: http://www.jvbd.org//text.asp?2021/58/3/183/321739

  Introduction Top

Ticks with a worldwide distribution are exclusively hematophagous ectoparasite. In addition to inflicting direct damage by their voracious blood feeding activities, ticks are ranked second after mosquitoes as the vector of different kind of pathogens to humans, domestic and wild animals[1]. In recent years, the number of cases of Kyasanur Forest Disease (KFD)[2], Crimean Congo Hemorrhagic fever (CCHF)[3],[4], Lyme disease[5] and Indian Tick Typhus (ITT)[6], Tick relapsing fever[7],[8] have been on the rise in India. These changes are also reported from USA[9], Europe[10] and Australia[11], largely due to spreading of tick vectors in new areas[12]. Tick control is the one of the key solutions to prevent the spreading of tick-borne diseases (TBDs). Currently, the tick control method is focused on the use of chemical acaricides on animals and on animals shed usually located adjacent to the humans and animal habitat. However, indiscriminate use of chemical acaricides led to the development and establishment of acaricide-resistant tick populations[13],[14]. These groups of chemicals also create environmental concerns and the effects on human health have been reported[15],[16],[17].

On broad literature analysis, fourteen major TBDs have been reported from India. The vector potential of ticks and epidemiological pattern of TTBDs are likely to change with the changing environmental conditions and the failure to manage tick infestations in sustainable manner is expected to impact TBDs scenarios significantly[18]. Among the existing tick management methods, immunization of hosts against targeted tick species is an environmentally friendly, cost-effective and easy to use approach to minimize the use of chemical acaricides for the management of TTBDs[19],[20]. One of the success stories linked with parasite management programme is the identification and establishment of Bm86 protein (tick midgut antigen) based commercial vaccines, TickGARD and Gavac, which were utilized in the integrated tick management program in Australia and in Latin American countries against Rhipicephalus microplus infestations[20],[21],[22].

Every year there is an increasing trend of tick-borne human disease outbreak being reported from India and this data are regularly updated by National Centre for Disease Control, New Delhi (compiled in [Figure 1]) and also in other parts of the world[10],[11]. Subsequently, the demand of anti-tick vaccine was increased many folds due to establishment of multi-acaricide resistant tick populations in different parts of the globe[23],[24],[25],[26],[27],[28],[29],[30],[31]. Development of acaricide resistant tick populations, increased public awareness towards food safety and non-sustainable nature of chemical-based tick management system, prompted an effective environment friendly approach involving cross-protective vaccines as a sustainable tick management option[32],[33],[34],[35].
Figure 1: Number of tick-borne human disease outbreaks per year

Click here to view

Accordingly, for vaccine development, the first challenge is the identification of antigen(s) which should be accessible to the host immune system and are conserved across the tick species. Various molecules were identified in different tick species viz., Bm86[36], Bm91[37], Bm95[38], Calreticulin[39], Subolesin[40], Ferritin-2[41],[42] Glutathione S-transferase[43], Aquaporin-1[44], Reprolysin [BrRm-MP4][45], Glutathione s-transferase mu (DmGstm1)[46], cystatin 2a (Racys2a)[47], but, none of these antigens provide significant level of immunity against multi-ticks infestations and tick-borne pathogens.

In India, initially, native tick proteins were used for experimental immunization of laboratory and natural hosts with some encouraging results[48],[49],[50],[51],[52],[53],[54],[55]. However, due to non-availability of funding support to most of the laboratories working on the subject and also due to non-availability of trained manpower on the area, the positive results obtained using native antigens could not be progressed further. Later, Azhahianambi et al[56] while working on Hyalomma anatolicum cloned and expressed recombinant Haa 86 (homolougus to R. microplus Bm86 gene) of H. anatolicum in E. coli and tested against challenge conditions. The efficacy of the immunogen against adults was 61.4-82.3 % and 47% against larvae of H. anatolicum. Subsequently, the post challenge efficacy of immunization of calves with recombinant Subolesin (rHa-SUB), Calreticulin (rHa-CRT) and Cathepsin L (rHa-CathL) were recorded 65.4%, 41.3% and 30.2% against H. anatolicum and 54.0%, 37.6% and 22.2%, respectively against R. microplus[57]. Further, the Ferritin2 (FER2) and Tropomysin (TPM) genes of H. anatolicum were cloned, expressed in E. coli system and tested. The efficacy of the antigens was 51.78% and 63.77%, respectively, against the challenge infestation[58]

In most of the laboratories the tick vaccine research is focused on single antigen immunization with limited or no cross protection. Another approach to enhance the efficacy of tick vaccines is use of a combination of protective antigens. It has been reported that combination of two tick/ pathogen antigens can significantly increase the vaccine efficacy[37],[38],[39],[40],[41],[42],[43],[44],[45],[46],[47],[59],[60],[61],[62],[63],[64],[65]. Some of the tick protective antigens producing long-lasting immunity can be used to prevent or reduce tick infestations and pathogen infection and transmission in domestic animals, wild reservoir hosts and humans[32].

Livestock population of India, Africa and other south Asian countries are infested with more than one tick species. Till date none of the identified antigens have been shown to have significant cross-protective efficacy, and cross-protective efficacy is one of the key characteristics of ticks for commercial exploitation. Hence, to develop sustainable solution to fight against multiple tick species, conserved multi antigen tick vaccine is the best possible option. The advantages of multi antigen tick vaccine over single antigen vaccines are:

  1. Immunity against different tick species/ stages can be achieved following immunization.
  2. Vaccines consisting of multi tick and tick transmitting pathogen antigens can block host pathogen multiplication cycle.
  3. The vaccines will be commercially viable and can be incorporated in integrated tick management system in countries where animals are infested with multiple tick species.

However, identification of ideal vaccine candidates targeting multiple physiological functions of ticks is most challenging. This review is focused on the updated status of development of multi-antigenic vaccine for the control of TTBDs.

Multi-antigen tick vaccines

In most countries, multi-tick infestations on animals are the major issues. A universal anti-tick vaccine that renders protection against multiple tick species is the commercial need posing a steep challenge before the scientific community working in this area. Majority of the literature addresses the single-antigen immunization protocol for development of prophylactic measures against ticks. However, no single-antigen vaccine is providing appreciable cross-protection to establish a commercially successful entity. Willadsen et al[66] and Parizi et al[60] are of the opinion that an anti-tick vaccine containing multiple antigens is more likely to be more effective than single antigen vaccine. Accordingly, focus has been directed to study the synergistic effect of multi-antigens tick vaccine which have different mechanisms of action to produce broad-spectrum effects on tick biology. In the process of identification of antigens, initially, RNAi technology was used to study the gene function, characterization of tick-pathogen interface, and to establish the feasibility and practicality of multi tick vaccine[67]. The salient observation of the experiments is compiled in [Table 1].
Table 1: Effects of multi genes RNAi studies on tick survival.

Click here to view

Chimeric vaccine

“Chimera” means combination of two different forms/ antigens. This technology is used to enhance the immunization efficacy or to develop new vaccine constructs targeting multiple functions. It was proposed that different tick antigens/epitopes can be expressed as a single protein which can elicit cross-protection against heterologous tick challenges. However, chimeric tick vaccine development is still in its early stage and few attempts were made involving vector and pathogen components.

BM95-MSP1a construct of chimeric protein using BM95 antigen of R. microplus and Major Surface Protein 1a (MSP1a) of Anaplasma marginale expressed in E. coli. This vaccine successfully induced significant antibody titers in rabbits[71]. Almazan et al[59] conducted vaccine trial using this vaccine in cattle and observed 54% reduction of R. microplus infestations and overall efficacy of 64%. A Subolesin-MSP1a chimeric construct of subolesin protein of R. microplus and Major Surface Protein 1a (MSP1a) of A. marginale expressed in E. coli. Vaccination of cattle showed 34% reduction of R. microplus infestation and overall efficacy of 81%[59]. The vaccination of cattle showed 8-fold reduction in infestation percent of animals[64] and vaccination of sheep reduced tick infestations by 63%. Weight of the female ticks was reduced by 32–55% compared to control ticks and Babesia bigemina seroprevalence was lowered by 30% in vaccinated cattle[72]. The Elongation Factor 1a (EF1a) - MSP1a construct involved EF1a of R. microplus and MSP1a of A. marginale protective epitopes were fused and vaccination of cattle resulted in 38% reduction of R. microplus infestation, 7% reduction in tick weight, 22% reduction in egg fertility and 38% overall efficacy against tick infestations[59]. Similarly, the Q41 Subolesin-Akirin construct consists of fused protective epitopes of Ixodes scapularis subolesin antigen and protective epitopes of Aedes albopictus akirin antigen. The Cysteine encoding nucleotides were inserted in between the selected subolesin and akirin epitopes to form disulphide bridges which helps in epitope conformation. Vaccination of mice with Q41 showed reduction of female mosquito survivability and fertility and overall, 99 % efficacy[61]. Lastly, the Q38 Subolesin-Akirin construct comprised of fused protective epitopes of I. scapularis subolesin (SUB) and protective epitopes of A. albopictus akirin antigens was developed. A GGGS amino acids spacer was introduced for better exposure of epitopes. Vaccination with Q38 reduced oviposition of both mosquitoes (28%) and sand flies (26%) fed on vaccinated mice[61]. The efficacy of Q38 vaccination against larvae of I. ricinus was 99.9% and 46.4% against D. reticulatus[72]. Contreras et al[76] conducted a vaccination study in the field condition using Q38 and showed promising results in controlling multiple tick species infesting roe deer (Capreolus capreolus) population in Spain. Unfortunately, except Q38 none of the above-mentioned chimeric constructs was tested in natural host against experimental challenge condition.

Cocktails of protein as vaccine

The vaccine formulation prepared using functionally different antigens with adjuvant is referred to as cocktails. This kind of vaccine is formulated to get broad spectrum activity or to reach the threshold level of protection where single antigen-based vaccines cannot achieve. The broad-spectrum vaccines are always a choice over the single species vaccine to address the multi-species infestations on animals in the field situation. A few immunization trials have been conducted using cocktails of protein and are listed [Table 2].
Table 2: The efficacy of cocktail vaccines reported

Click here to view

Recently, Ndawula Jr. et al[65] worked on to develop the broad-spectrum cocktail vaccine contains recombinant GST (glutathione S-transferases) proteins of different tick species. They expressed the targeted proteins from five different tick species viz., R. appendiculatus, R. decoloratus, R. microplus, Amblyomma variegatum and Haemaphysalis longicornis in E. coli as rGST-Ra, rGST-Rd, rGST-Rm, rGST-Av, rGST-Hl. Immunization of animals using cocktails of all the recombinants showed significant humoral immune response and stronger cross-recognition of sera produced against rGST-Rd and rGST-Av to heterologous rGST compared to sera produced against rGST-Ra, rGST-Hl or rGST-Rm. Moreover, sera against all the rGSTs cross-recognize R. sanguineus crude egg protein extracts, however, sera against rGST-Ra and rGST-Rd showed highest reactivity. Accordingly, the cocktail vaccine of rGST-Rd and rGST-Av was formulated, which showed 35% reduction of female R. sanguineus infestations on challenged rabbits. The antigen selection is the crucial part in cocktail vaccine design. Though, it is reported that one individual antigen may interfere with the protection conferred by the other antigen and this can lead to reduction of overall protection of the cocktail vaccine, a systematic approach using combinations of different technologies like in-silico analysis using online software’s like VaxiJen needs to be followed for constituting cocktail-antigen vaccines.


In contrast to cocktail vaccine, multiple antigens prepared separately and injected separately as a vaccine may elicit better response and chances of interference of one antigen with other can be avoided. Only three reports of such vaccination study are available. A preliminary study of co-vaccination with Bm86 and Bm91 induced significantly higher immunogenicity than vaccination with Bm86 alone but no synergy was observed[37]. McKenna et al[77] purified BMA7 and Bm86 from R. microplus and following co-vaccination of cattle elicited strong Anti-Bm86 (4200-94000) and Anti-BmA7(1600-22000) antibody titer resulted in reduction of egg masses to 2.09–6.04 g/day of the feeding ticks. Co-vaccination of yeast expressed Bm86 and E. coli expressed Subolesin antigens in cattle showed 97% reduction against challenge infestations[78]. Most of the co-vaccination trials were conducted in small number of animals. The dose rate, species of animals, type of tick antigens, adjuvants and site of vaccine administration needs to be standardized before going for co-vaccination using a greater number of animals.

Multi antigen oral vaccine

Oral vaccines against ticks are still in their early stage of development [Table 3]. Oral vaccines have several advantages over other vaccine delivery routes, viz., no need of adjuvants, easy to administer, speed of vaccine delivery is very quick, trained personnel is not required for administration of vaccine, no pain during vaccine administration, less risk of contamination or infection at the injection site[79],[80]. Multi-antigens based oral vaccine formulations can be used for the immunization of wild life reservoirs against tick infestations to prevent tick borne pathogens from overpass the inter species barrier and cause disease in animals and humans[81]. Two experiments were conducted, and the results are compiled in [Table 3].
Table 3: Compiled results of oral vaccine against tick infestations.

Click here to view

The orally administered SUB-MSP1a is the first evidence of the protective capacity of the membrane antigen administered orally to immunize hosts and supported the use of membrane-bound antigens in vaccine formulations. Oral vaccines also have some disadvantages such as low immunogenicity and antigen stability after immunization. To counteract these disadvantages vaccines formulations with selected combinations of pre tested antigens, antigen stability enhancers in acidic stomach environment and vaccine delivery systems needs to be standardized before initiation of vaccine trial[79],[80],[81],[82]. The protection observed using oral SUB+IV vaccine provided new path for future experiments but a number of variables are to be optimized for conducting oral vaccine trials in larger number of animals.

Future strategies

In the past 20 years, research on development of immunoprophylactic measure against ticks has grown rapidly along with technological development. But during the same period, several new TTBDs have been emerged worldwid[69],[70]. To counter the emerging tread of TTBDs, there is a need to adopt technologies which are ecofriendly, less expensive, easy to use and have broader activity.

Omics approach

de la Fuente et al[85] opined that identification of new target antigens is possible through tick-pathogen interaction study using systems biology approach that helps in selection of the best targets to control tick infestations and pathogen transmission. Vaccinomics involved integrated product of “omics” technologies such as transcriptomics, proteomics, immunogenomics and with systems biology and bioinformatics for the development of next-generation vaccines. With the advancement of new technologies, it has been hypothesized that for designing effective multi antigens vaccine targeting against both ticks and the pathogens it transmits, a combination of new tools viz., interactomics with proteomics, transcriptomics, metabolomics, regulomics together with Big Data analytic techniques are to be used strategically[86]. Omics-based approach of selection of different adjuvants has already been utilized in human vaccine research[87],[88] so, there is a strong need of omics-based selection of adjuvants in tick vaccinology research too.

Lipid nano particle (LNP)-mRNA vaccines

The mRNA encoding two genes of the Powassan virus, an emerging tick-borne virus, captured in lipid nanoparticles and reported that one dose of the vaccine was enough to induce robust immunity. No viremia or significant anamnestic antibody response was observed following POWV challenge. This immunization protocol produced cross protecting antibodies against other flavivirus viz., OHFV (Omsk hemorrhagic fever virus), KFDV (Kyasanur forest disease virus) and AHF (Alkhurma hemorrhagic fever virus)[89]. The mRNA encoding multiple selected tick and/or pathogen genes can be incorporated in lipid nanoparticles and this multi tick and/or pathogen antigen LNP-mRNA vaccine can provide significant protection against TTBDs.

Viral vector vaccines

The gene sequence of protective tick antigen can be incorporated in the genome of vector-borne virus and such viruses are inoculated in to host either through oral / nasal /subcutaneous routes. The inoculated virus produces tick protein along with viral proteins and these tick proteins induce immunity in host against tick antigens. Viral vector vaccines have several advantages viz.,

  1. Induce humoral as well as cell mediated immunity.
  2. No need of adjuvant.
  3. High efficiency of gene transduction.

Oral vaccination with subolesin expressing vaccinia virus inhibited tick infestation by 52% compared to control vaccination with vaccinia virus and reduced uptake of Borrelia burgdorferi by 34% among the surviving ticks that fed to repletion. A 40% reduction in transmission of B. burgdorferi to uninfected vaccinated mice in comparison to controls was recorded. The results of these studies suggest that subolesin incorporation in live vector vaccine may have potential as a component of a reservoir targeted vaccine, to decrease B. burgdorferi, Babesia and Anaplasma species infections in their natural hosts[90].

Live parasite vaccines

The gene sequence of tick antigen can be incorporated in genomes of low virulent live parasites. These low virulent parasites are given as a vaccine through oral/subcutaneous routes. The parasite in host body can express tick proteins and can induce protection against ticks as well as parasites. Oldiges et al[91] developed live Babesia bovis strain S74-T3B expressing tick antigen, glutathione S-transferase from Haemaphysalis longicornis (HlGST). This type of low virulent Babesia can induce protective antibodies against babesiosis and anti-GST antibodies in host. Immunization trial induced detectable anti-glutathione S-transferase antibodies and showed reduced tick size and fecundity of H. longicornis and R. microplus feeding on experimentally immunized animals. This type of low virulent tick-borne pathogens expressing more than two tick protective antigens can be used for conferring protection against both ticks and pathogens.

New vaccine delivery systems

Amongst the existing antigen delivery systems, nano and micro-particles-based delivery are the most promising since it has better efficiency to enhance the cross-presentation of the antigen, and activate both innate and adaptive immune systems[92]. Recently, Chopenko et al[93] developed an effective nano vaccine against tick borne encephalitis virus. The chimeric antigen is consisting of tick-borne encephalitis (TBE) E protein domain III of virus and OmpFporin of Yersinia pseudotuberculosis incorporated in TI-complexes [TI-complexes are self-organized from mixture of Cucumaria japonica triterpene glycosides and marine macrophytes cholesterol and monogalactosyldiacyl glycerol (MGDG)]. The MGDG plays role in lipid matrix for subunit protein antigen interrupted in TI-complexes. Micro viscosity of MGDG was shown to influence the conformation and immunogenicity of protein antigen and provided a 60% protection. Virus-like immune stimulating complexes (ISCOMs) usually considered as “gold standard” delivery systems, which has greater adjuvant efficacy and stability than liposomes and aluminum. Kostetsky et al[94] reported that the TI-complexes demonstrated high adjuvant efficiency in relation to OmpFporin isolated from Y. pseudotuberculosis in comparison to ISCOM and Freund’s complete adjuvant. Similarly, tick multi antigens can be incorporated in TI-complexes for effective delivery of the antigens. Other strategies like multi antigenic epitope-based DNA vaccines, self-assembling nanoparticle-based vaccines, gold nanoparticle carrying antigen vaccines also can be adopted for multi antigens vaccine development

A significant gap does exist in multi antigens tick vaccines research. For example;

  1. Worldwide different laboratories have identified a number of antigens with limited or no study on cross protective potential of the identified antigens;
  2. Limited studies have been conducted to understand protein-protein interaction which is very crucial while combining multiple antigens;
  3. A number of adjuvants are commercially available to potentiate and maintain immune response for longer duration, but again selection of adjuvants in multi antigens format is very tricky and has to be worked out experimentally;
  4. The different research groups working on tick vaccine are using different animal models mostly rabbits, guinea pigs, mice, rats etc. and in many cases the data generated in these models can not be repeated in the original hosts.

To overcome these drawbacks following strategies can be taken up:

  1. Currently only few tick databases are available Bmi-GI[95], Cattle TickBase[96] and these are not enough to screen the tick vaccine candidates. There should be country specific tick databases which can be accessible by all the tick research groups worldwide;
  2. High throughput discovery and characterization of several tick antigens and pathogen derived protective antigens can be studied using intelligent big data analytic techniques (de la Fuente et al[97]);
  3. Functional genomics studies of tick antigens and vector interface using large scale or high throughput experimental methodologies together with statistical, computational and bioinformatics analysis of the results[98] are essential;
  4. The animal models should be selected based on the type of natural hosts. Initial experiments can be conducted on suitable animal models but in vivo immunization of natural hosts and challenge with suitable stage of the tick species is essential to prove potential of any combination of multiple antigens;
  5. There should be country wise specific bodies mandated to monitor TTBDs status and the consortium of these bodies should exchange important data maintaining IPR portfolio to solve the present problem and to manage future challenges.

In conclusion, multi antigens tick vaccine development is still in the early stage. But the results obtained in the last 5–10 years have demonstrated the possibilities of this approach for the management of TTBDs. A significant research shift towards omics approach has helped in understanding the functional aspects of the targets to be used in multi-targets vaccine formulation.

  Acknowledgements Top

The authors are grateful to the Indian Council of Agricultural Research, New Delhi for funding through the National Agricultural Science Fund [Grant number NASF/ABA-6015/2016-17/357 and NFBSFARA/BSA-4004/2013-14].

Conflict of interest: None

  References Top

de la Fuente J, Estrada-Pena A, Venzal JM, Kocan KM, Sonen-shine DE. Overview: ticks as vectors of pathogens that cause disease in humans and animals. Front Biosci 2008; 13(13): 693–846.  Back to cited text no. 1
Sadanandane C, Elango A, Marja N, Sasidharan PV, Raju KH, Jambulingam P. An outbreak of Kyasanur forest disease in the Wayanad and Malappuram districts of Kerala, India. Ticks Tick Borne Dis 2017; 8(1): 25–30.  Back to cited text no. 2
Yadav PD, Patil DY, Shete AM, Kokate P, Goyal P, Jadhav S, Sinha S, Zawar D, Sharma SK, Kapil A, Sharma DK. Nosocomial infection of CCHF among health care workers in Rajasthan, India. BMC Infect Dis 2016; 16(1): 624.  Back to cited text no. 3
Gurav YK, Yadav PD, Deostwar AR, Dhruwey VS, Shete AM, Chaubal GY. Sero-survey of Crimean Congo Haemorrhagic fever (CCHF) among high risk group during 2011 and 2013 CCHF outbreak in Gujarat, India. Indian J Applied Res 2014; 4(12): 430.  Back to cited text no. 4
Jairath V, Sehrawat M, Jindal N, Jain VK, Aggarwal P. Lyme disease in Haryana, India. Indian J Dermatol Venereol Leprol 2014; 80(4): 320.  Back to cited text no. 5
Tirumala S, Behera B, Jawalkar S, Mishra PK, Patalay PV et al.. Indian tick typhus presenting as Purpura fulminans. Indian J Crit Care Med 2014; 18(7): 476.  Back to cited text no. 6
Aher AR, Shah H, Rastogi V, Tukaram PK, Choudhury RC. A case report of relapsing fever. Indian J Pathol Microbiol 2008; 51(2):292.  Back to cited text no. 7
Veena S, Seema V, Babu R. Borreliosis: Recurrent fever due to spirochetes. Ann Trop Med Public Health 2013; 6(4):482.  Back to cited text no. 8
Centre for Disease Control, lyme disease statistics, USA. [Internet]. 2020. Available from:https://www.cdc.gov/lyme/stats/ tables.html?CDC_AA_refVal=https%3A%2F%2Fwww.cdc.go v%2Flyme%2Fstats%2Fchartstables%2Freportedcases_state-locality.html (Accessed on)  Back to cited text no. 9
El-Sayed A, Kamel M. Climatic changes and their role in emergence and re-emergence of diseases. Environ Sci Pollut Res 2020:1–17.  Back to cited text no. 10
Dehhaghi M, Kazemi Shariat Panahi H, Holmes EC, Hudson BJ, Schloeffel R, Guillemin GJ. Human tick-borne diseases in Australia. Front Cell Infect Microbiol 2019: 9: 3.  Back to cited text no. 11
Chakraborty S, Andrade FC, Ghosh S, Uelmen J, Ruiz MO. Historical expansion of Kyasanur forest disease in India from 1957 to 2017: a retrospective analysis. GeoHealth. 2019; 3(2): 44–55.  Back to cited text no. 12
Chigure GM, Sharma AK, Kumar S, Fular A, Sagar SV, Nagar G, et al. Role of metabolic enzymes in conferring resistance to synthetic pyrethroids, organophosphates, and phenylpyrazole compounds in Rhipicephalus microplus. Int J Acarol 2018; 44(1): 28–34.  Back to cited text no. 13
Nandi A, Sagar SV, Chigure G, Fular A, Sharma AK, Nagar G, et al. Determination and validation of discriminating concentration of ivermectin against Rhipicephalus microplus. Vet Parasitol 2018; 250: 30–4.  Back to cited text no. 14
Kaushik A, Sharma HR, Jain S, Dawra J, Kaushik CP. Pesticide pollution of river Ghaggar in Haryana, India, Environ Monit Assess 2010; 160(1-4): 61.  Back to cited text no. 15
Jayaraj R, Megha P, Sreedev P. Organochlorine pesticides, their toxic effects on living organisms and their fate in the environment. Interdiscip Toxicol 2016; 9(3-4): 90–100.  Back to cited text no. 16
Anju A, Ravi S P, Bechan S. Water pollution with special reference to pesticide contamination in India. Water Resour Prot 2010; 02(05): 432–448.  Back to cited text no. 17
Ghosh S, Azhahianambi P, de la Fuente J. Control of ticks of ruminants, with special emphasis on livestock farming systems in India: present and future possibilities for integrated control— a review. Exp Appl Acarol 2006; 40(1): 49–66.  Back to cited text no. 18
Contreras Rojo M, Kasija PD, Merino O, de la Cruz Hernandez NI, Gortazar C, De La Fuente J. Oral vaccination with a formulation combining Rhipicephalus microplus Subolesin with heat inactivated Mycobacterium bovis reduces tick infestations in cattle. Front Cell Infect Microbiol 2019; 9: 45.  Back to cited text no. 19
Valle MR, Mèndez L, Valdez M, Redondo M, Espinosa CM, Vargas M, et al. Integrated control of Boophilus microplus ticks in Cuba based on vaccination with the anti-tick vaccine Gavac TM. Exp Appl Acarol 2004; 34(3-4): 375–82.  Back to cited text no. 20
de la Fuente J, Almazán C, Canales M, de la Lastra JM, Kocan KM, Willadsen P. A ten-year review of commercial vaccine performance for control of tick infestations on cattle. Anim Health Res Rev 2007; 8(1): 23–8.  Back to cited text no. 21
Willadsen P. Anti-tick vaccines. In: Bowman, A.S and Nuttal. P.A. (eds) Ticks: biology, disease and control. Cambridge (UK): Cambridge University Press; 2008. 424-446.  Back to cited text no. 22
Li AY, Davey RB, Miller RJ, George JE. Detection and characterization of amitraz resistance in the southern cattle tick, Boophilus microplus (Acari: Ixodidae). J Med Entomol 2004; 41(2): 193–200.  Back to cited text no. 23
Perez-Cogollo LC, Rodriguez-Vivas RI, Ramirez-Cruz GT, Miller RJ. First report of the cattle tick Rhipicephalus microplus resistant to ivermectin in Mexico. Vet Parasitol 2010; 168(1-2): 165–9.  Back to cited text no. 24
Lopez-Arias A, Villar-Argaiz D, Chaparro-Gutierrez JJ, Miller RJ, De Leon AA. Reduced efficacy of commercial acaricides against populations of resistant cattle tick Rhipicephalus microplus from two municipalities of Antioquia, Colombia. Environ health insights 2014; 8: EHI–S16006.  Back to cited text no. 25
Ghosh S, Kumar R, Nagar G, Kumar S, Sharma AK, Srivastava A, et al. Survey of acaricides resistance status of Rhipiciphalus (Boophilus) microplus collected from selected places of Bihar, an eastern state of India. Ticks Tick-Borne Dis 2015; 6(5): 668–75.  Back to cited text no. 26
Nandi A, Singh H, Singh NK. Esterase and glutathione S-transferase levels associated with synthetic pyrethroid resistance in Hyalomma anatolicum and Rhipicephalus microplus ticks from Punjab, India. Exp Appl Acarol 2015; 66(1): 141–57.  Back to cited text no. 27
Nandi A, Singh H, Singh NK, Rath SS. Fenvalerate resistance status in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) from Punjab, India. J Parasit Dis 2016; 40(3): 694–8.  Back to cited text no. 28
Upadhaya D, Kumar B, Kumar S, Sharma AK, Fular A, et al. Characterization of acaricide resistance in Rhipicephalus microplus populations infesting cattle in northeastern India and assessment of local plant extracts for tick management. Vet Parasitol 2020; 277: 109011.  Back to cited text no. 29
Vudriko P, Okwee-Acai J, Tayebwa DS, Byaruhanga J, Kakooza S, Wampande E, et al. Emergence of multi-acaricide resistant Rhipicephalus ticks and its implication on chemical tick control in Uganda. Parasite Vectors 2016; 9(1): 4.  Back to cited text no. 30
Sagar SV, Saini K, Sharma AK, Kumar S, Kumar R, Fular A, et al. Acaricide resistance in Rhipicephalus microplus collected from selected districts of Madhya Pradesh, Uttar Pradesh and Punjab states of India. Trop Anim Health Prod 2020; 52(2): 611–8.  Back to cited text no. 31
De la Fuente J, Contreras M. Tick vaccines: current status and future directions. Expert Rev Vaccines 2015; 14(10): 1367–76.  Back to cited text no. 32
Lambertz C, Chongkasikit N, Jittapalapong S, Gauly M. Immune response of Bos indicus cattle against the anti-tick antigen Bm91 derived from local Rhipicephalus (Boophilus) microplus ticks and its effect on tick reproduction under natural infestation. J Parasitol Res2012: 907607.  Back to cited text no. 33
De la Fuente J, Contreras M, Estrada-Peña A, Cabezas-Cruz A. Targeting a global health problem: vaccine design and challenges for the control of tick-borne diseases. Vaccine 2017; 35(38): 5089–94.  Back to cited text no. 34
De la Fuente J. Controlling ticks and tick-borne diseases… looking forward. Ticks Tick Borne Dis 2018; 9(5): 1354–7.  Back to cited text no. 35
Willadsen P, Bird P, Cobon GS, Hungerford J. Commercialisation of a recombinant vaccine against Boophilus microplus. Parasitology 1995; 110(S1): 43–50.  Back to cited text no. 36
Willadsen P, Smith D, Cobon G, McKenna RV. Comparative vaccination of cattle against Boophilus microplus with recombinant antigen Bm86 alone or in combination with recombinant Bm91. Parasite Immunol 1996; 18(5): 241–6.  Back to cited text no. 37
García-García JC, Montero C, Redondo M, Vargas M, Canales M, Boue O, et al. Control of ticks resistant to immunization with Bm86 in cattle vaccinated with the recombinant antigen Bm95 isolated from the cattle tick, Boophilus microplus. Vaccine 2000; 18(21): 2275–87.  Back to cited text no. 38
Parizi LF, Rech H, Ferreira CA, Imamura S, Ohashi K, Onuma M, et al. Comparative immunogenicity of Haemaphysalis longicornis and Rhipicephalus (Boophilus) microplus calreticulins. Vet Parasitol 2009; 164(2-4): 282–90.  Back to cited text no. 39
Kasaija PD, Contreras M, Kabi F, Mugerwa S, de la Fuente J. Vaccination with recombinant subolesin antigens provides cross-tick species protection in Bos indicus and crossbred cattle in Uganda. Vaccines 2020; 8(2): 319.  Back to cited text no. 40
Hajdusek O, Almazán C, Loosova G, Villar M, Canales M, Grubhoffer L, et al. Characterization of ferritin 2 for the control of tick infestations. Vaccine. 2010; 28(17): 2993–8.  Back to cited text no. 41
Githaka NW, Konnai S, Isezaki M, Goto S, Xavier MA, Fujisawa S, et al. Identification and functional analysis of ferritin 2 from the Taiga tick Ixodes persulcatus Schulze. Ticks Tick Borne Dis 2020; 11(6): 101547.  Back to cited text no. 42
Parizi LF, Utiumi KU, Imamura S, Onuma M, Ohashi K, Masuda A, et al. Cross immunity with Haemaphysalis longicornis glutathione S-transferase reduces an experimental Rhipicephalus (Boophilus) microplus infestation. Exp Parasitol 2011; 127(1): 113–8.  Back to cited text no. 43
Hussein HE, Scoles GA, Ueti MW, Suarez CE, Adham FK, Guerrero FD, et al. Targeted silencing of the Aquaporin 2 gene of Rhipicephalus (Boophilus) microplus reduces tick fitness. Parasite Vector 2015; 8(1): 618.  Back to cited text no. 44
Ali A, Parizi LF, Guizzo MG, Tirloni L, Seixas A, da Silva Vaz Jr I, Termignoni C. Immunoprotective potential of a Rhipicephalus (Boophilus) microplus metalloprotease. Vet Parasitol 2015; 207(1-2): 107–14.  Back to cited text no. 45
Song R, Li M, Fan X, Hu Z, Wu L, Li Y, et al. Caracterization of glutathione S-transferase of Dermacantor marginatus and effect of the recombinant antigen as a potential anti-tick vaccine. Vet Parasitol 2020; 279: 109043.  Back to cited text no. 46
Parizi LF, Rangel CK, Sabadin GA, Saggin BF, Kiio I, Xavier MA, et al. Rhipicephalus microplus cystatin as a potential cross-protective tick vaccine against Rhipicephalus appendiculatus. Ticks Tick-Borne Dis 2020; 11(3): 101378.  Back to cited text no. 47
Parmar A, Grewal AS, Dhillon P. Immunological cross-reactivity between salivary gland proteins of Hyalomma anatolicum anatolicum and Boophilus microplus ticks. Vet Immunol Immunopathol 1996; 51(3-4): 345–52.  Back to cited text no. 48
Ghosh S, Khan MH. Identification of tick antigen immunogenic in calves. Anim Sci J 1997; 12: 249–52.  Back to cited text no. 49
Ghosh S, Khan MH, Gupta SC. Immunization of rabbits against Hyalomma anatolicum anatolicum using homogenates from unfed immature ticks. Indian J Exp Biol1998; 36(2): 167–70.  Back to cited text no. 50
Sangwan AK, Banerjee DP, Sangwan N. Immunization of cattle with nymphal Hyalomma anatolicum anatolicum extracts: effects on tick biology. Trop Anim Health Prod 1998; 30(2): 97–106.  Back to cited text no. 51
Ghosh S, Khan MH, Ahmed N. Cross-bred cattle protected against Hyalomma anatolicum anatolicum by larval antigens purified by immunoaffinity chromatography. Trop Anim Health Prod 1999; 31(5): 263–73.  Back to cited text no. 52
Das G, Ghosh S, Khan MH, Sharma JK. Immunization of crossbred cattle against Hyalomma anatolicum anatolicum by purified antigens. Exp Appl Acarol 2000; 24(8): 645–59.  Back to cited text no. 53
Sharma, J. K., Ghosh, S., Khan, M. H. and Das, G. Immunoprotective efficacy of 39 kDa purified nymphal antigen of Hyalomma anatolicum. Tropical Animal Health and Production 2001; 33(2): 103–113.  Back to cited text no. 54
Ghosh S, Singh NK, Das G. Assessment of duration of immunity in crossbred cattle immunized with glycoproteins isolated from Hyalomma anatolicum anatolicum and Boophilus microplus. Parasitol Res 2005; 95(5): 319–26.  Back to cited text no. 55
Azhahianambi P, De La Fuente J, Suryanarayana VV, Ghosh S. Cloning, expression and immunoprotective efficacy of rHaa86, the homologue of the Bm86 tick vaccine antigen, from Hyalomma anatolicum anatolicum. Parasite Immunol 2009; 31(3): 111–22.  Back to cited text no. 56
Kumar B, Manjunathachar HV, Nagar G, Ravikumar G, de la Fuente J, Saravanan BC, Ghosh S. Functional characterization of candidate antigens of Hyalomma anatolicum and evaluation of its cross-protective efficacy against Rhipicephalus microplus. Vaccine 2017; 35(42): 5682–92.  Back to cited text no. 57
Manjunathachar HV, Kumar B, Saravanan BC, Choudhary S, Mohanty AK, Nagar G, et al. Identification and characterization of vaccine candidates against Hyalomma anatolicum—Vector of Crimean Congo haemorrhagic fever virus. Transbound Emerg Dis 2019; 66(1): 422–34.  Back to cited text no. 58
Almazán C, Moreno-Cantú O, Moreno-Cid JA, Galindo RC, Canales M, Villar M, et al. Control of tick infestations in cattle vaccinated with bacterial membranes containing surface-exposed tick protective antigens. Vaccine 2012; 30(2): 265–72.  Back to cited text no. 59
Parizi LF, Githaka NW, Logullo C, Konnai S, Masuda A, Ohashi K, et al. The quest for a universal vaccine against ticks: cross-immunity insights. Vet J 2012; 194(2): 158–65.  Back to cited text no. 60
Moreno-Cid JA, de la Lastra JM, Villar M, Jiménez M, Pinal R, Estrada-Peña A, et al. SUB/AKR Vaccine Study Group. Control of multiple arthropod vector infestations with subolesin/akirin vaccines. Vaccine 2013; 31(8): 1187–96.  Back to cited text no. 61
Parizi LF, Reck Jr J, Oldiges DP, Guizzo MG, Seixas A, Logullo C, et al. Multi-antigenic vaccine against the cattle tick Rhipicephalus (Boophilus) microplus: a field evaluation. Vaccine 2012; 30(48): 6912–7.  Back to cited text no. 62
Contreras M, Moreno-Cid JA, Domingos A, Canales M, Díez-Delgado I, de la Lastra JM, et al. Bacterial membranes enhance the immunogenicity and protective capacity of the surface exposed tick Subolesin-Anaplasma marginale MSP1a chimeric antigen. Ticks Tick-Borne Dis 2015; 6(6): 820–8.  Back to cited text no. 63
Torina A, Moreno-Cid JA, Blanda V, de Mera IG, de la Lastra JM, Scimeca S, et al.. Control of tick infestations and pathogen prevalence in cattle and sheep farms vaccinated with the recombinant Subolesin-Major Surface Protein 1a chimeric antigen. Parasites Vectors 2014; 7(1): 10.  Back to cited text no. 64
Ndawula Jr C, Sabadin GA, Parizi LF, da Silva Vaz Jr I. Constituting a glutathione S-transferase-cocktail vaccine against tick infestation. Vaccine 2019; 37(14): 1918–27.  Back to cited text no. 65
Willadsen P. Antigen cocktails: valid hypothesis or unsubstantiated hope?. Trends Parasitol 2008; 24(4): 164–7.  Back to cited text no. 66
de la Fuente J, Kocan KM, Almazán C, Blouin EF. RNA interference for the study and genetic manipulation of ticks. Trends Parasitol 2007; 23(9): 427–33.  Back to cited text no. 67
de la Fuente J, Almazán C, Naranjo V, Blouin EF, Kocan KM. Synergistic effect of silencing the expression of tick protective antigens 4D8 and Rs86 in Rhipicephalus sanguineus by RNA interference. Parasitol Res 2006; 99(2): 108–13.  Back to cited text no. 68
Smith A, Guo X, de la Fuente J, Naranjo V, Kocan KM, Kaufman WR. The impact of RNA interference of the subolesin and voraxin genes in male Amblyomma hebraeum (Acari: Ixodidae) on female engorgement and oviposition. Exp Appl Acarol 2009; 47(1): 71–86.  Back to cited text no. 69
Rahman M, Saiful Islam M, You M. Impact of subolesin and cystatin knockdown by RNA interference in adult female Haemaphysalis longicornis (Acari: Ixodidae) on blood engorgement and reproduction. Insects 2018; 9(2): 39.  Back to cited text no. 70
Canales M, Almazán C, de la Lastra JM, de la Fuente J. Anaplasma marginale major surface protein 1a directs cell surface display of tick BM95 immunogenic peptides on Escherichia coli. J Biotechnol 2008; 135(4): 326–32.  Back to cited text no. 71
Contreras M, de la Fuente J. Control of Ixodes ricinus and Dermacentor reticulatus tick infestations in rabbits vaccinated with the Q38 Subolesin/Akirin chimera. Vaccine 2016; 34(27): 3010–3.  Back to cited text no. 72
Almazán C, Kocan KM, Blouin EF, de la Fuente J. Vaccination with recombinant tick antigens for the control of Ixodes scapularis adult infestations. Vaccine 2005; 23(46-47): 5294–8.  Back to cited text no. 73
Imamura S, Namangala B, Tajima T, Tembo ME, Yasuda J, Ohashi K, et al. Two serine protease inhibitors (serpins) that induce a bovine protective immune response against Rhipicephalus appendiculatus ticks. Vaccine 2006; 24(13): 2230–7.  Back to cited text no. 74
Imamura S, Konnai S, da Silva Vaz IJ, Yamada S, Nakajima C, Ito Y, et al. Effects of anti-tick cocktail vaccine against Rhipicephalus appendiculatus. Jpn J Vet Res 2008; 56(2): 85–98.  Back to cited text no. 75
Contreras M, San José C, Estrada-Peña A, Talavera V, Rayas E, León CI, Núñez JL, et al. Control of tick infestations in wild roe deer (Capreolus capreolus) vaccinated with the Q38 Subolesin/ Akirin chimera. Vaccine 2020; 38(41): 6450–4.  Back to cited text no. 76
McKenna RV, Riding GA, Jarmey JM, Pearson RD, Willadsen P. Vaccination of cattle against the Boophilus microplus using a mucin-like membrane glycoprotein. Parasite Immunol 1998; 20(7): 325–36.  Back to cited text no. 77
Schetters TP, Jansen T. Vaccine against Rhipicephalus ticks 2017. United States patent US 9,579,369.  Back to cited text no. 78
Wang S, Liu H, Zhang X, Qian F. Intranasal and oral vaccination with protein-based antigens: advantages, challenges and formulation strategies. Protein Cell 2015; 6(7): 480–503.  Back to cited text no. 79
Lawan A, Jesse FF, Idris UH, Odhah MN, Arsalan M, Muhammad NA, et al. Mucosal and systemic responses of immunogenic vaccines candidates against enteric Escherichia coli infections in ruminants: a review. Microb Pathog 2018; 117: 175–83.  Back to cited text no. 80
Gortazar C, Reperant LA, Kuiken T, de la Fuente J, Boadella M, Martínez-Lopez B, et al. Crossing the interspecies barrier: opening the door to zoonotic pathogens. PLoS pathog 2014; 10(6): e1004129.  Back to cited text no. 81
Fry T, Dalen KV, Hurley J, Nash P. Mucosal adjuvants to improve wildlife rabies vaccination. J Wildl Dis 2012; 48(4): 1042–6.  Back to cited text no. 82
Kosoy OI, Lambert AJ, Hawkinson DJ, Pastula DM, Goldsmith CS, Hunt DC, Staples JE. Novel thogoto virus associated with febrile illness and death, United States, 2014. Emerg infect dis 2015; 21(5): 760.  Back to cited text no. 83
Kernif T, Leulmi H, Raoult D, Parola P. Emerging Tick Borne Bacterial Pathogens. Emerg Infect 2016: 295–310.  Back to cited text no. 84
de la Fuente J, Kopácek P, Lew Tabor A, Maritz Olivier C. Strategies for new and improved vaccines against ticks and tick borne diseases. Parasite Immunol 2016; 38(12): 754–69.  Back to cited text no. 85
Artigas-Jerónimo S, De La Fuente J, Villar M. Interactomics and tick vaccine development: new directions for the control of tick-borne diseases. Expert Rev Proteome 2018; 15(8): 627–35.  Back to cited text no. 86
Burny W, Callegaro A, Bechtold V, Clement F, Delhaye S, Fissette L, et al. Different adjuvants induce common innate pathways that are associated with enhanced adaptive responses against a model antigen in humans. Front Immunol 2017; 8: 943.  Back to cited text no. 87
Harandi AM. Systems analysis of human vaccine adjuvants. In Seminars in immunology. Academic Press; 2018; 39: 30–34.  Back to cited text no. 88
VanBlargan LA, Himansu S, Foreman BM, Ebel GD, Pierson TC, Diamond MS. An mRNA vaccine protects mice against multiple tick-transmitted flavivirus infections.Cell Rep 2018; 25(12): 3382–92.  Back to cited text no. 89
Bensaci M, Bhattacharya D, Clark R, Hu LT. Oral vaccination with vaccinia virus expressing the tick antigen subolesin inhibits tick feeding and transmission of Borrelia burgdorferi. Vaccine 2012; 30(42): 6040–6.  Back to cited text no. 90
Oldiges DP, Laughery JM, Tagliari NJ, Leite Filho RV, Davis WC, da Silva Vaz Jr I, et al. Transfected Babesia bovis expressing a tick GST as a live vector vaccine. PLoS Negl Trop Dis 2016; 10(12): e0005152.  Back to cited text no. 91
Tel J, Sittig SP, Blom RA, Cruz LJ, Schreibelt G, Figdor CG, de Vries IJ. Targeting uptake receptors on human plasmacytoid dendritic cells triggers antigen cross-presentation and robust type I IFN secretion. J Immunol 2013; 191(10): 5005–12.  Back to cited text no. 92
Chopenko N, Mazeika A, Davydova L, Stenkova A, Leonova G, Kostetsky E, et al. Effectivity of nanovaccine against tick-borne encephalitis. In J Phys Conference Series 2018; Vol. 1092: 012020.  Back to cited text no. 93
Kostetsky EY, Sanina NM, Mazeika AN, Tsybulsky AV, Vorobyeva NS, Shnyrov VL. Tubular immunostimulating complex based on cucumarioside A 2-2 and monogalactosyldiacylglycerol from marine macrophytes. J Nanobiotechnol 2011; 9(1): 35.  Back to cited text no. 94
Guerrero FD, Miller RJ, Rousseau ME, Sunkara S, Quacken-bush J, Lee Y, Nene V. BmiGI: a database of cDNAs expressed in Boophilus microplus, the tropical/southern cattle tick. Insect Biochem Mol Biol 2005; 35(6): 585–95.  Back to cited text no. 95
Bellgard MI, Moolhuijzen PM, Guerrero FD, Schibeci D, Rodriguez-Valle M, Peterson DG, Dowd SE, Barrero R, Hunter A, Miller RJ, Lew-Tabor AE. CattleTickBase: an integrated Internet-based bioinformatics resource for Rhipicephalus (Boophilus) microplus. Int. J. Parasitol 2012; 42(2): 161–9.  Back to cited text no. 96
de La Fuente J, Villar M, Estrada-Peña A, Olivas JA. High throughput discovery and characterization of tick and pathogen vaccine protective antigens using vaccinomics with intelligent Big Data analytic techniques. Expert Rev Vaccines 2018; 17(7): 569–76.  Back to cited text no. 97
Valle MR, Guerrero FD. Anti-tick vaccines in the omics era. Front Biosci 2018 Jan;10: 122–36  Back to cited text no. 98


  [Figure 1]

  [Table 1], [Table 2], [Table 3]


    Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
    Access Statistics
    Email Alert *
    Add to My List *
* Registration required (free)  

  In this article
Article Figures
Article Tables

 Article Access Statistics
    PDF Downloaded343    
    Comments [Add]    

Recommend this journal