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Ticks and tick-borne diseases are a major constraint on livestock farming in many developing countries, which has a huge impact on their economies. Genomic information is becoming more abundant for many of the species involved, which if exploited successfully could be used to develop new control strategies. Here, we review the genomic resources that are now available and discuss how this information is currently being harnessed or can be used in the future to explore the complex interplay that occurs between livestock hosts, tick vectors and tick-borne pathogens.

Overwhelming evidence has accumulated of the effectiveness of immunization with live attenuated vaccines to control tick-borne diseases of livestock. Despite several disadvantages, vaccination with live attenuated organisms against tropical theileriosis, babesiosis and possibly heartwater constitutes one of the most cost-effective intervention strategies. Although great advances have been made through genomics and proteomics research, this has not yet translated into effective non-living vaccines. As a result, there is a continuing necessity to use available live vaccines in tick and tick-borne disease-control strategies adapted to conditions prevailing in many parts of the world.

Spotted fever group rickettsioses are emerging and reemerging infectious diseases, some of which are life-threatening. In order to understand how dendritic cells (DCs) contribute to the host resistance or susceptibility to rickettsial diseases, we first characterized the in vitro interaction of rickettsiae with bone marrow-derived DCs (BMDCs) from resistant C57BL/6 (B6) and susceptible C3H/HeN (C3H) mice. In contrast to the exclusively cytosolic localization within endothelial cells, rickettsiae efficiently entered and localized in both phagosomes and cytosol of BMDCs from both mouse strains. Rickettsia conorii-infected BMDCs from resistant mice harbored higher bacterial loads compared to C3H mice. R. conorii infection induced maturation of BMDCs from both mouse strains as judged by upregulated expression of classical major histocompatibility complex (MHC) and costimulatory molecules. Compared to C3H counterparts, B6 BMDCs exhibited higher expression levels of MHC class II and higher interleukin-12 (IL-12) p40 production upon rickettsial infection and were more potent in priming naïve CD4(+) T cells to produce gamma interferon. In vitro DC infection and T-cell priming studies suggested a delayed CD4(+) T-cell activation and suppressed Th1/Th2 cell development in C3H mice. The suppressive CD4(+) T-cell responses seen in C3H mice were associated with a high frequency of Foxp3(+) T regulatory cells promoted by syngeneic R. conorii-infected BMDCs in the presence of IL-2. These data suggest that rickettsiae can target DCs to stimulate a protective type 1 response in resistant hosts but suppressive adaptive immunity in susceptible hosts.

Vaccines based on recombinant Bm86 gut antigen from Boophilus microplus are a useful component of integrated control strategies against B. microplus infestations of cattle. The capacity of such vaccines to control heterologous infestations by two African tick species was investigated. The mean weight of engorged female ticks and mean egg mass per tick were significantly reduced in B. decoloratus infestations, but there was no effect of the vaccine against adult Rhipicephalus appendiculatus. We cloned, sequenced and expressed two Bm86 homologues (Bd86) from B. decoloratus. Amino acid sequence identity between Bd86 homologues (Bd86-1 and Bd86-2) and Bm86 was 86% and 85%, respectively, compared to 93% identity between the variants. Native Bd86 protein in B. decoloratus tick mid-gut sections and recombinant Bd86-1 reacted strongly with sera from TickGARD vaccinated cattle. TickGARD can therefore protect against a heterologous tick species with multiple antigen sequences. Epitope mapping using sera from TickGARD-vaccinated cattle identified two linear peptides conserved between the Bd86 homologues and Bm86. These epitopes represent candidate synthetic peptide vaccines for control of Boophilus spp. and the pathogens transmitted by these tick vectors.

The prevalence and incidences of tick-borne diseases (TBDs) infections in cattle were studied in the western-Kenya highlands. Serological tests, thick-blood and lymph-node smears were used to quantify TBDs during cross-sectional and longitudinal studies. Four hundred and eight and 192 (wet season) and 114 and 46 (dry season) cattle were bled for serology in the rural and peri-urban areas, respectively. Seroprevalences differed significantly between the two areas (p<0.05). The cattle were monitored for 23 months (259,923 and 126,273 cattle days in the rural and peri-urban areas, respectively) from September 1996 to August 1998. The incidences of babesiosis, anaplasmosis and theileriosis were 0.42%, 4.64% and 4.92% and 1.45%, 32.11% and 39.05% in the rural and peri-urban areas, respectively. The difference in the incidence were significantly different for anaplasmosis and theileriosis (p<0.05). The difference in disease frequencies in two areas in close proximity shows the importance of monitoring disease events in order to understand and advice farmers in different production and farming systems appropriately.

This study investigated the seroprevalence of antibodies against Anaplasma phagocytophilum in Berlin/Brandenburg, north-eastern Germany. During 1994-2001, 422 sera from patients with proven tick-exposure (specimens with antibodies against Borrelia burgdorferi) were compared with 249 control sera. Using indirect fluorescent antibody testing, significantly more positive samples were detected among Borrelia antibody-positive specimens (4.5%, 95% CI 2.5-6.5%) than among controls (1.2%, 95% CI 0.5-1.9%; p < 0.05). While six (2.2%, 95% CI 1.3-3.1%) samples were positive among Borrelia antibody-positive sera between 1994 and 1997, 13 (8.7%, 95% CI 6.9-10.5%) were positive between 1998 and 2001 (p < 0.01), indicating an uneven annual seroprevalence.

Ticks are distributed worldwide and impact human and animal health, as well as food animal production. Control of ticks has been primarily by application of acaricides, which has resulted in selection of resistant ticks and environmental pollution. Vaccines have been shown to be a feasible tick control method that offers a cost-effective, environmentally friendly alternative to chemical control. However, identification of tick-protective antigens remains the limiting step in vaccine development. Tick antigens exposed naturally to the host during tick feeding and those concealed have both shown promise as candidate vaccine antigens. Development of vaccines against multiple tick species may be possible using highly conserved tick-protective antigens or by antigens showing immune cross-reaction to different tick species. Vaccines made from a combination of key protective antigens may greatly enhance vaccine efficacy. Preliminary studies have suggested the possibility of vaccine strategies directed toward both tick control and the blocking of pathogen transmission. Characterization of the tick genomes will have a great impact on the discovery of new protective antigens. The future of research directed toward tick vaccine development is exciting because of new and emerging technologies for gene discovery, and vaccine formulation and delivery.

Tick-borne pathogen transmission is dependent upon tick number per host and the physical and temporal distribution of each feeding stage. Age-related acquired immunity to tick and pathogen may also be important but has received less attention. In this study we evaluate which of these parameters has the greatest impact on Great Island Virus (GIV) transmission between Ixodes uriae ticks and common guillemots (Uria aalge). The study system is well suited to investigate age-related effects because the guillemot population is naturally divided into 2 groups, older breeding and younger pre-breeding adult birds. The physical distribution and timing of adult and nymphal tick feeding was similar for both guillemot age groups. However, breeding birds were parasitized by significantly more ticks (mainly nymphs). Calculations based on tick number predict virus prevalence should be higher in ticks that have fed on breeding rather than pre-breeding birds. However, empirical evidence indicates the reverse. Protective acquired immunity to GIV infection may be the reason why GIV prevalence is actually significantly lower in ticks that have fed on breeders. Far more breeding (74%) than pre-breeding (12%) guillemots had antibodies that neutralized 1 or more GIV strains. Estimates of the force of infection support the view that pre-breeding birds experience higher rates of virus infection than breeding birds. The results indicate age-related acquired immunity is a key factor in GIV transmission and highlight the need to consider age-related effects and host immunity when undertaking quantitative studies of tick-borne pathogen transmission.

Ornithodoros erraticus is an argasid tick that can transmit severe diseases such as human relapsing fever and African swine fever. In southern Europe O. erraticus lives in close association with swine on free-range pig farms. Application of acaricides for the eradication of O. erraticus from pig farms is inefficient. This is the reason why we tried to develop an anti-O. erraticus vaccine as alternative method of control. Accordingly, we were prompted to investigate the protective possibilities of a midgut membrane extract from the parasite (GME) that has not been studied hitherto. Administration of the GME with Freund's adjuvants (FAs) to pigs and mice induced a protective response able to kill 80% of the immature forms of the parasite in the first 72 h post-feeding and to reduce the fecundity of females by more than 50%. The action of the vaccine is the result of damage to the midgut wall of the argasid, and, in mice, it has been shown that this damage is mediated by activation of the complement system. In pigs, the administration of GME with alum, instead of with FAs, reduced the degree of protection. The protective antigens of the GME were expressed by all the developmental stages examined and are probably proteins from the luminal membrane of midgut epithelial cells. These antigens were seen to be more abundant in recently fed parasites than in fasting specimens, suggesting that their expression is induced after blood ingestion.

A study using a structured questionnaire was conducted to assess the perception of cattle farmers of the efficacy of East Coast fever (ECF) immunization in southern Zambia. One hundred and seventy-nine farmers from five districts in southern Zambia were interviewed. The majority of farmers (85%) perceived ECF immunization as being very effective and about half of them (51.4%) preferred immunization to other ECF control strategies. The study showed that the number of calves immunized was strongly associated with the farmer's perception of the benefits of immunization. There was no association between the number of calves immunized and the number of veterinary assistants in a given district or their transport situation. Overall mortality in ECF-immunized calves from various causes stood at 4.2%. Based on farmers' reports, the majority of these deaths seemed to have been caused by anaplasmosis, another tick-borne disease, and might have resulted from relaxation of tick control after ECF immunization. The reasons identified by farmers for not immunizing their animals included failure by immunizing teams to reach certain areas, not having calves of immunization age, and lack of money. These findings provide valuable information on how livestock farmers perceive and adopt new animal disease control strategies and the information could be useful in their planning and implementation.

Ticks are dangerous blood-sucking ectoparasites of medical and veterinary importance. They serve as reservoirs and vectors pathogens and cause direct skin damage by feeding. Tick saliva contains pharmacologically active molecules directed against host's immune system and effector pathways. Ticks feeding induce hosts' inflammatory and immunological reactions of host comprise both innate and acquired immunity.

cDNA expression library immunization (ELI) and analysis of expressed sequenced tags (EST) in a mouse model of tick infestations was used to identified cDNA clones that affected I. scapularis. Three protective antigens against larval tick infestations, 4F8, with homology to a nucleotidase, and 4D8 and 4E6 of unknown function, were selected for further characterization. All three antigens were expressed in all I. scapularis stages and localized in adult tick tissues. 4D8 was shown to be conserved in six other tick species. Based on immunization trials with synthetic polypeptides against larvae and nymphs and on artificial feeding experiments of adults, these antigens, especially 4D8, appear to be good candidates for continued development of a vaccine for control of tick infestations and may be useful in a formulation to target multiple species of ticks.

Antigens prepared from ovine granulocytes and tick cells infected with ovine strains of Anaplasma phagocytophilum, the causative agent of tick-borne fever, were tested in respect of their suitability for the assay of antibodies in ovine sera by enzyme-linked immunosorbent assay (ELISA) and by indirect immunofluorescent antibody test (IFAT). Antigens prepared from tick cells were as sensitive and specific as those expressed in ovine granulocytes for the detection of specific antibodies by ELISA, but they failed to react in the IFAT with immune sera obtained from sheep previously infected with ovine strains of A. phagocytophilum.

Methods currently used to control Ixodes scapularis ticks rely principally on acaricidal applications which suffer from a number of limitations. Recently, host vaccination against ticks has been shown to be a promising alternative tick control method. In tick salivary glands, numerous genes are induced during the feeding process. Many of these newly expressed proteins are secreted in tick saliva and may play a role in modulating host immune responses and pathogen transmission. We have performed suppression subtraction hybridization to identify unique I. scapularis salary gland proteins specifically expressed during engorgement. We have cloned and sequenced ten unique salivary gland-associated cDNAs that are up-regulated during feeding. The protein products of these genes represent potential vaccine candidates for use in the control of ticks and to prevent transmission of tick-borne diseases.

Babesiosis is a tick-borne disease of cattle caused by Babesia bigemina and Babesia bovis and is transmitted by the tick vector Boophilus microplus. In this study, we investigate B. bigemina infection regarding the clinical infection, T cell distribution, and cytokine profile during the acute phase of an experimental infection in pregnant cows.

Boophilus microplus is an important vector of bovine disease agents having a major economic impact on cattle production in many tropical and subtropical countries. Components of tick saliva that enable ticks to feed may also facilitate establishment of tick-borne pathogens in the vertebrate host. It has been suggested that acquired resistance against molecules in tick saliva could inhibit parasite transmission, and there is increasing evidence to support this hypothesis. The effect of immune resistance to B. microplus on the incidence of tick-transmitted pathogens was the focus of this experiment. Groups of four dairy cows were injected with antigen extracts of tick salivary glands, midgut, adjuvant only, or PBS, prior to a grazing period in a pasture in Thailand where ticks are abundant and babesiosis is enzootic. These animals were then observed for evidence of babesiosis throughout the rainy season. A reduction in the incidence of clinical babesiosis was observed among cattle immunized with salivary gland preparations compared to nonimmunized controls (P < 0.05). Immunization with midgut or adjuvant only both resulted in a slight reduction in observed disease compared to the same negative control group. B. bigemina was detected in fewer ticks (24.43%) collected from salivary gland-immunized cattle than those collected from the remaining groups (> or =44.57%). These results indicated that immunization with salivary gland antigens could affect pathogen transmission and appears promising for control of tick-borne diseases of cattle.

In testing paired serum samples from 40 consecutive cases of African tick bite fever, we detected diagnostic antibodies against spotted fever group rickettsiae in 45% of the patients by immunofluorescence assay (IFA) and in 100% of the patients by Western blotting (WB) (P < 0.01). A specific diagnosis of Rickettsia africae infection could be established in 15% of the patients by IFA and in 73% of the patients by a combination of WB and cross-adsorption assays (P < 0.01).

Immunization against the protozoan Theileria parva by infection and treatment has proved to be very efficient for the control of East Coast fever, an acute and often-fatal lymphoproliferative tick-borne disease of cattle in Eastern, Central and Southern Africa. The immunizing dose of live T. parva sporozoites used in this method is usually determined by in vivo titration. An alternative in vitro method of quantification of sporozoites in whole tick-derived stabilates is proposed. The method consists of incubating serially diluted T. parva stabilates with bovine peripheral blood lymphocytes, the host cell that is infected naturally. Allowing the cultures to incubate undisturbed for the full cultivation period (10 days) reduced the variability among replicate titrations. Fungal contaminations were avoided by centrifuging stabilates at 400 g prior to the incubation, which did not precipitate sporozoites significantly. Fungistatics, Nystatin and Flucytosine, did not appear to interfere with the in vitro development of T. parva but their effect on fungal growth was limited. In vitro titration data were compared to in vivo infection data for 2 stabilates. In vitro titration of T. parva sporozoites should allow more ethical and efficient research on the preparation and storage of T. parva tick-derived stabilates.

Between August and October 2000, a cross-sectional study was conducted in smallholder dairy farms in Mbale District, Uganda to assess the prevalence of ticks and tick-borne diseases under different grazing systems and agro-ecological zones and understand the circumstances under which farmers operated. A questionnaire was administered to obtain information on dairy farm circumstances and practices. A total of 102 farms were visited and sera and ticks were collected from 478 animals. Sero-prevalence of tick-borne diseases was determined using an enzyme-linked immunoassay. Acaricides were used indiscriminately but the intensity of their use varied with the grazing system and zone. Cattle from different farms mixed for various reasons. During the dry seasons farmers have to get additional fodder from outside their farms that can result in importation of ticks. The prevalence of ticks and serum antibodies to tick-borne infections differed across the grazing systems and zones. The highest serum antibody prevalence (>60%) was recorded in the lowland zone under the free range and tethering grazing systems. The lowest tick challenge and serum antibody levels (<50%) were recorded in the midland and upland zones under a zero-grazing system. These findings suggest that endemic stability to East Coast Fever, babesiosis and anaplasmosis is most likely to have existed in the lowland zone, particularly, under the tethering and free-range grazing systems. Also, endemic stability for babesiosis existed in the upland zones. Endemic instability for East Coast Fever existed in the midland and upland zones. These structured observational studies are instrumental in planning of control strategies for ticks and tick borne diseases since production systems and the cattle population at high risk of the diseases in the district have been identified.

Ticks are of vast medical and veterinary public health importance due to direct damage caused by feeding and their roles in transmitting well known and emerging infectious agents. Ticks and tick-borne pathogens stimulate the immune system of the host. Those immune interactions are of importance in tick biology, pathogen transmission and control of ticks and tick-borne diseases. Both innate and specific acquired immune defenses are involved in the responses of vertebrate hosts to infestation. Ticks have evolved countermeasures to circumvent host immune defenses. This review addresses the immunobiology of the tick-host interface from the perspectives of the pharmacology of tick saliva; relationship of tick saliva to pathogen transmission; host immune responses to infestation; tick modulation of host immune defences; and genomic/proteomic strategies for studying tick salivary gland molecules.

When feeding on vertebrate host ticks (ectoparasitic arthropods and potential vectors of bacterial, rickettsial, protozoal, and viral diseases) induce both innate and specific acquired host-immune reactions as part of anti-tick defenses. In a resistant host immune defense can lead to reduced tick viability, sometimes resulting in tick death. Tick responds to the host immune attack by secreting saliva containing pharmacologically active molecules and modulating host immune response. Tick saliva-effected immunomodulation at the attachment site facilitates both tick feeding and enhances the success of transmission of pathogens from tick into the host. On the other hand, host immunization with antigens from tick saliva can induce anti-tick resistance and is seen to be able to induce immunity against pathogens transmitted by ticks. Many pharmacological properties of saliva described in ticks are shared widely among other blood-feeding arthropods.

With a view to determine which tick species bite humans most frequently in the province of Soria (Spain) and to know whether these species act as vectors of Rickettsia conorii and/or Borrelia burgdorferi, we analysed the presence of antibodies against salivary proteins of several tick species and of antibodies against R. conorii and B. burgdorferi sensu lato in 102 samples of serum. The sera were collected from 63 patients who were treated for tick bites at the health services of the Soria Health Area. Thirty-nine percent of the patients displayed antibodies against Dermacentor marginatus; 21% against Ixodes ricinus, and 6% against Rhipicephalus sanguineus. IgM antibodies against R. conorii were found in 15 patients, but none of the patients included in the study displayed IgG/IgM Abs against B. burgdorferi sensu lato. In 30% of the patients bitten by I. ricinus and in 24% bitten by D. marginatus, antibodies against R. conorii were found. None of the patients who had been bitten by R. sanguineus expressed antibodies against this pathogen. The results obtained here suggest that D. marginatus and I. ricinus are the main vectors of the spotted fever group rickettsiae in our environment and that the role of R. sanguineus in the transmission of these rickettsiae is debatable.

The risk of acquiring North Asian tick typhus (infection by Rickettsia sibirica) during travel to regions of Asia where this disease is endemic is unknown. We investigated prospectively 13 paleontologists on expedition to Mongolia. Four paleontologists had acute illness characterised by fever, rash, headache, and lymphadenopathy. All had IgM and IgG antibodies to R sibirica. Paleontologists with no illness and people who went on expeditions in other parts of the world did not have antibodies to R sibirica. Only two of the four infected persons were aware of tick bites. Travellers to regions endemic for R sibirica are at risk of contracting North Asian tick typhus even in the absence of recognised tick-bites.

Anaplasma phagocytophilum infection in sheep is characterized by an immune suppression as indicated by impaired antibody response, reduced lymphocyte response and reduced oxidative burst. The effect of A. phagocytophilum infection on leucocyte populations, especially lymphocytes, was therefore investigated in six sheep experimentally infected with A. phagocytophilum, and compared with leucocyte populations from control animals.To investigate the ability of the infection to interfere with the cellular and humoral responses to specific antigens, the animals were vaccinated with commercial vaccines at the time of experimental infection, and monitored for 56 days. There were reduced percentages of gammadelta T-cells and CD4+ T-cells in peripheral blood of infected animals throughout the study period, and these cell populations showed a down-regulation of CD25 expression; while there was a relative increase in CD8+ T-cells. The reduction in CD25+ gammadelta T-cells involved a subpopulation of WC1+ gammadelta T-cells. During the first 2 weeks of the study there were reduced percentages of B-cells and leukocytes expressing MHC II and CD11b, though this decrease changed to a relative increase later in the study. The relative reductions in leucocyte populations corresponded with the observed leucopenia during the first 3 weeks post-infection, which involved lymphocyte, neutrophil and eosinophil subsets [Vet. Immunol. Immunopathol. 86 (2002) 183]. There was a reduced expression of CD11b and CD14 on granulocytes during the first 2 weeks of the study, which corresponded with the previously reported leucopenia involving neutrophils and eosinophils. Antibody responses to vaccines, lymphocyte in vitro proliferative responses to antigens and mitogens, and in vitro IFN-gamma responses to antigens were reduced up to 4 weeks after infection.

Ticks are economically important ectoparasites of domestic and wild animals and are considered to be second worldwide to mosquitoes as vectors of human pathogens. Current control methods for ticks, based primarily on the use of acaricides, have had limited efficacy in the reduction of tick infestations and the use of acaricides is often accompanied by serious drawbacks, including selection of acaricide-resistant ticks and environmental contamination. Development of improved vaccines against tick infestations offers a cost-effective and environmentally sound control method. Commercial vaccines currently marketed for control of cattle ticks have been effective in field studies when used in concert with integrated control strategies. However, new antigens are needed to increase the efficacy of tick vaccines. Although a limited number of protective antigens against tick infestations have been identified and characterized, discovery of new antigens remains the limiting step for improving the efficacy of tick vaccines. Recent technologies developed for gene discovery, including expression library immunization and evaluation of expressed sequence tags, show promise for rapid, systematic and global antigen screening and should provide a comprehensive approach to selection of candidate vaccine antigens. Design of future tick vaccines should target multiple tick species, as well as interfere with the transmission of pathogens.

Demand for live trivalent tick fever vaccine containing Babesia bovis, Babesia bigemina and Anaplasma centrale produced by the Department of Primary Industries, Queensland, has increased from less than 10,000 doses in 1988 to 500,000 doses in 2001. This paper describes a series of trials aimed at overcoming certain constraints to obtain B. bigemina parasitised erythrocytes (PEs) on a large enough scale from infected splenectomised calves to meet the demand. Passage through a series of splenectomised calves failed to increase the yield per calf but we showed that the dose rate of infected cells could be reduced from the long-time standard of 1x10(7) to 2.5x10(6) without affecting immunogenicity and still leaving a safety margin of at least 50-fold for infectivity. This change quadrupled the potential yield of doses per calf and allowed the DPI to meet the increased demand for B bigemina in vaccine. Due to the high cost and limited availability of suitable, health tested donors, calves previously infected with B. bovis or A. centrale were used to provide B. bigemina organisms but the practice resulted in red cell agglutination in some batches of prepared vaccine. A trial is described where B. bigemina-infected red cells were washed by centrifugation to remove agglutinating antibodies. Washing had no effect on parasite viability and this method is now in routine use in the production of trivalent vaccine.

A field isolate of Theileria annulata (Uzbek strain) was obtained from calves infected by Hyalomma anatolicum ticks collected from an endemic region in Uzbekistan. Schizont-infected bovine cells that had been established and propagated in cell culture were examined for attenuation both in vivo, by inoculating cells from various passages into calves, and in vitro for metalloproteinase activity. During serial subcultivation a gradual reduction in virulence and in enzyme activity in cells infected with the Uzbek strain were observed. Complete attenuation of the Uzbek isolate was obtained at about passage 80, and only traces of proteolysis were detected in gelatin substrate gels. In contrast, there was no direct correlation between virulence and enzyme levels in an Israeli strain. While schizonts of the Israeli strain were completely attenuated at passage 80, proteolysis in the substrate gels was detected up to passage 197. Solid immunity was observed in calves immunized with attenuated T. annulata schizonts of the Uzbek strain upon challenge with the homologous H. excavatum sporozoites. For a strain to be used for vaccine production, it appears that animal inoculation still remains the most reliable method to assess the degree of attenuation and protection.

Objective: To evaluate the sequential changes and to estimate the frequencies of abnormalities in some commonly measured biological variables in patients with African tick bite fever (ATBF), an emerging spotted fever group (SFG) rickettsiosis in international travelers to rural sub-Saharan Africa., Methods: A study was done of hemoglobin, total leukocyte count, absolute lymphocyte count, blood platelet count and serum levels of C-reactive protein (S-CRP), alanine aminotransferase (S-ALAT), aspartate aminotransferase, lactic dehydrogenase, gamma-glutamyl transferase, alkaline phosphatase, bilirubin, sodium and creatinine during the first two weeks of illness and prior to the institution of antirickettsial therapy in 108 patients with travel-associated ATBF., Results: There were significant falls in mean total leukocyte count, mean absolute lymphocyte count, and mean platelet count, and significant increases in mean S-CRP and S-ALAT. During the first ten days of illness, elevated S-CRP, lymphopenia and elevated S-ALAT were detected in 91.7%, 73.3% and 40.7% of patients, respectively. Most abnormalities were mild. For 55 patients who underwent both S-CRP and absolute lymphocyte count determination, at least one parameter was abnormal in 52 (94.5%) patients., Conclusions: The sequential changes in many biological parameters during the acute phase of ATBF mimic those reported in other SFG rickettsioses. Mild abnormalities are frequent, with increased S-CRP and lymphopenia being the two most consistent findings.

Information regarding the inflammatory response in African tick-bite fever (ATBF), an emerging spotted-fever-group rickettsiosis, in international travelers to sub-Saharan Africa, is scarce. Plasma/serum levels of von Willebrand factor (vWF), soluble (s) E-selectin, tumor necrosis factor-alpha, interleukin (IL)-6, interferon-gamma, IL-10, IL-13, IL-8, RANTES, macrophage inflammatory protein-1alpha, and C-reactive protein were studied, at both first presentation and follow-up, in 15 patients with travel-associated ATBF and in 14 healthy travelers who served as control subjects. Our main and novel findings are the following: (1) patients with ATBF had increased levels of vWF and sE-selectin, with a subsequent decrease at follow-up; (2) with the exception of IFN-gamma, levels of cytokines and chemokines were also increased in these patients at the first presentation; and (3) IL-10 and IL-13 tended to increase during follow-up, whereas most of the inflammatory cytokines decreased. The induction of these mediators and the balance between them may be critical both for the regulation of inflammation and for protective immunity in ATBF.

A total of 1,335 archived human sera collected in 1985 from an area in Japan where a tick-borne disease is endemic were examined by indirect immunofluorescence antibody test (IFAT) to estimate seroprevalence against three serologically distinct types of Babesia microti-like parasites; namely, Hobetsu, Kobe, and U.S. types. Eighteen sera (1.3%) were found to be IFAT-positive (titer 1:100-1:6,400), of which 14 and three were ascertained by Western blot analysis to be positive against the Hobetsu and Kobe types, respectively. In addition, four sera showed an IFAT titer of 1:100 against the U.S. type, but they appeared to be false-positive because they were cross-reactive against the Hobetsu and Kobe types, and also because a U.S.-type parasite has not been found in Japan. Our results suggest that human babesiosis in Japan occurred prior to the discovery of the index case in 1999 and that the infections were caused mainly by Hobetsu-type parasites.

Heartwater is controlled by frequent application of acaricides, which is costly, creates endemic instability and has the potential of contaminating the environment. The live blood vaccine currently available has limitations because it is laborious and inconvenient to use, difficult to standardise and can transmit other blood-borne pathogens. The UF/USAID/SADC Heartwater Research Project has conducted research on the development of two types of vaccine for heartwater. The first-generation inactivated vaccine has been intensively tested in the laboratory and subsequently field tested in four southern African countries. It protects cattle, sheep and goats against mortality from heartwater challenge. It can be modified to incorporate any Ehrlichia ruminantium strain to provide protection from field challenge. The second-generation DNA vaccine containing genes encoding immunogenic E. ruminantium proteins has been developed and evaluated in the mouse model as well as in cattle and sheep. The use of improved vaccines against heartwater would have a positive impact on livestock farming in sub-Saharan Africa and the Caribbean and could be used to control the spread of heartwater if it were to be introduced into regions such as the United States.

Friesian cattle were immunized with two inoculations of anti-tick Bm86 (Tick-GARD) vaccine and were challenged 30 or 90 d later with Boophilus annulatus larvae derived from 1.2 g of eggs. No nymphs or adult ticks were found on the immunized cattle during four weeks after challenge. Repeated infestations (2 to 4) with larvae on three other calves during a period of 160 and 390 d after the immunization did not result in development of nymphal and adult stages. In control, non-immunized cattle infested with corresponding batches of larvae 1380 to 4653 replete adult female ticks were collected. Larvae issued from Babesia bovis-infected female ticks transmitted the infection to Bm86-immunized cattle, but the progeny of B. bigemina-infected females did not. Since B. bigemina is transmitted exclusively by nymphal stages of Bo. annulatus these results support the observation that immunity induced by Bm86 affects the larval stage of this tick.

Human granulocytic ehrlichiosis (HGE), a tick-borne zoonosis, is caused by an obligatory intragranulocytic bacterium, the HGE agent, a strain of Anaplasma phagocytophila. The equine model of HGE is considered valuable in understanding pathogenic and immune mechanisms of HGE. In the present study, cytokine mRNA expression by peripheral blood leukocytes (PBLs) in horses was examined during the course of infection by intravenous inoculation of A. phagocytophila or by allowing feeding by infected ticks. The p44 genes encoding the major outer membrane protein P44s of A. phagocytophila were detected by PCR in PBLs of all four horses from 4 to 20 days postexposure. During the 20-day infection period, interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNF-alpha) mRNA expression was upregulated in PBLs of all four horses, and IL-8 mRNA expression was upregulated in three horses. Gamma interferon, IL-10, and IL-12 p35 mRNAs were weakly expressed in only one horse each. IL-2, IL-4, IL-6, and IL-12 p40 mRNA expression, however, could not be detected in the PBLs of any of the four horses. These results suggest that IL-1beta, TNF-alpha, and IL-8 generation during A. phagocytophila infection has a primary role in HGE pathogenesis and immunomodulation.

A comparative study of human sera was conducted to determine which purified preparations of 11 recombinant antigens of Borrelia burgdorferi sensu stricto were diagnostically most important in enzyme-linked immunosorbent assays (ELISAs). To assess sensitivity, 20 serum samples obtained 1-6 weeks after onset of illness from 20 persons who had physician-diagnosed erythema migrans (EM) were tested for IgM and IgG antibodies. In tests for IgM antibody, seropositivity of > or = 25% was recorded when ELISAs had separate preparations of protein (p) 37, p41-G, outer-surface protein (Osp) C, OspE, OspF or VlsE antigens. Sera reacted most frequently (80% positive) with VlsE antigen in analyses for IgG antibodies. When results of both class-specific assays were considered for VlsE, OspC or OspF, 90% of the EM cases were serologically confirmed. Results of specificity testing with a further 59 sera from persons who had syphilis, louse-borne relapsing fever, oral infections, rheumatoid arthritis or human granulocytic ehrlichiosis and 28 normal sera indicated no false positive reactions when VlsE antigen was used in tests for IgM antibody. One of the 11 louse-borne relapsing fever sera cross-reacted with VlsE antigen in tests for IgG antibodies. Minor cross-reactivity also occurred when p37, OspC, OspE or OspF antigens were used. Overall, VlsE was the most suitable antigen for laboratory diagnosis of Lyme borreliosis during the early weeks of B. burgdorferi infection because of its high sensitivity and specificity.

Enzyme-linked immunosorbent assay (ELISA) was used to investigate the antibody responses of control sheep, and sheep naturally exposed to Ixodes ricinus Linné (Acari: Ixodidae) ticks, to salivary gland extract (SGE) proteins of partially fed, adult I. ricinus. Comparisons between responses of control sheep and naturally infested sheep by Western blot analysis suggested that variations in IgG responses of I. ricinus-exposed sheep were mostly associated with specific responses to I. ricinus SGE antigens. Sheep IgG responses were positively related to the numbers of adult ticks feeding per sheep at the time samples were collected, were greater during the spring than the autumn periods of I. ricinus activity and were inversely related to sheep resistance to ticks measured by the weights of nymphal I. ricinus that engorged on the sheep. These findings suggest that sheep lose their resistance to ticks due to polarization of a Th1 type response to some tick antigens towards a Th2 type response when sheep are exposed to high, natural tick infestations, or to seasonal conditions of relative nutritional stress. Potential consequences for the epidemiology of tick-borne diseases are discussed.

To determine whether a unique group of clinical and laboratory manifestations characterize certain major deer tick-transmitted human pathogens in North America, we compared the symptoms, short-term complications, and laboratory test results of New England residents who became ill due to > or =1 of these pathogens. Patients completed a uniformly structured questionnaire and submitted blood samples for serologic and polymerase chain reaction (PCR) testing after developing symptoms of Lyme disease, human babesiosis, or human granulocytic ehrlichiosis (HGE). Complete blood count with thin blood smear, PCR, and immunoglobulin M antibody tests helped differentiate the acute manifestations of these diseases. Physicians should consider use of tests designed to diagnose babesiosis and HGE in patients with Lyme disease who experience a prolonged flulike illness that fails to respond to appropriate antiborrelial therapy.

African tick-bite fever, caused by Rickettsia africae, is the most common tick-borne rickettsiosis in sub-Saharan Africa. Mediterranean spotted fever due to Rickettsia conorii also occurs in the region but is more prevalent in Mediterranean countries. Using microimmunofluorescence, we compared the development of immunoglobulin G (IgG) and IgM titers in 48 patients with African tick-bite fever and 48 patients with Mediterranean spotted fever. Doxycycline treatment within 7 days from the onset of disease significantly prevented the development of antibodies to R. africae. In patients with African tick-bite fever, the median times to seroconversion with IgG and IgM were 28 and 25 days, respectively, after the onset of symptoms. These were significantly longer by a median of 6 days for IgG and 9 days for IgM than the times for seroconversion in patients with Mediterranean spotted fever (P < 10(-2)). We recommend that sera collected 4 weeks after the onset of signs of patients with suspected African tick-bite fever should be used for the definitive serological diagnosis of R. africae infections.

With the development and licensure of a recombinant vaccine for the tick-borne infection Lyme disease, more attention has been paid to other vaccines that have been used or are being developed for the prevention of other tick-borne infections. This review highlights vaccine information for Lyme borreliosis, tick-borne encephalitis (TBE), Rocky Mountain spotted fever, tularaemia, Query (Q) fever, Kyasanur Forest disease (KFD) and tick paralysis. Additionally, discussion on the use of immunisation against the tick itself is included which not only can decrease veterinary tick burdens but may also decrease the transmission of arthropod-transmitted diseases.

Inoculation of sheep with Ehrlichia (previously Cowdria) ruminantium which has been cultivated in mammalian endothelial cell cultures is almost always followed by a severe clinical reaction, whereas inoculation of the agent cultivated in tick cell lines usually does not provoke a clinical response, but may result in seroconversion and/or protection against subsequent challenge with virulent stabilates. A quantitative, real-time PCR assay was developed to determine the kinetics of infection (rickettsaemia) in sheep inoculated with tick cell- and mammalian cell-derived E. ruminantium (Gardel isolate). The method and initial results are described, and the significance of the findings is discussed in relation to the clinical responses of the sheep.

Ehrlichia (previously Cowdria) ruminantium, the pathogen which causes heart-water in domestic and wild ruminants, can now be propagated in cell lines from one vector (Amblyomma variegatum) and five non-vector (Ixodes scapularis, I. ricinus, Boophilus decoloratus, B. microplus and Rhipicephalus appendiculatus) tick species. E. ruminantium isolates from West and South Africa and the Caribbean vary in their cell line preference, growth patterns and immunogenic capability. In laboratory trials, certain combinations of tick cell line and E. ruminantium isolate were highly immunogenic in sheep. These trial vaccines were grown under specific in vitro conditions and administered as a single intravenous dose of freshly harvested whole, live culture. Following immunisation and subsequent exposure to virulent E. ruminantium, protected sheep showed no clinical response and a range of serological responses.

Rickettsia africae is the causative agent of African tick bite fever (ATBF), an acute febrile illness frequently accompanied by inoculation eschars, regional lymphadenitis, myalgia and severe headache. Recently, ATBF has been recognized as an emerging health problem for international travellers to rural sub-Saharan Africa. To estimate the incidence, risk factors for and proportion of symptomatic cases of travel-associated R. africae infection, we performed a seroepidemiological study of 152 first-time Norwegian travellers to rural areas in sub-Equatorial Africa. Seropositivity was based on the detection of specific antibodies to R. africae in microimmunofluorescence and/or Western blotting assays. Thirteen (8.6%) travellers were seropositive to R. africae. Eight (62%) seropositive travellers reported symptoms consistent with ATBF; of these, 2 had received antirickettsial therapy. Using multiple logistic regression, the following factors were found to be significantly associated with seropositivity: hunting as the purpose of travel [odds ratio (OR) 10.1; 95% confidence interval (CI) 1.5-69; p=0.019] and stay in rural areas of > 7 d (OR 6.0; 95% CI 1.5-24; p=0.012). This first seroepidemiological study on travel-associated R. africae infection suggests that the infection may be common in international travellers to rural sub-Saharan Africa but that most cases are asymptomatic or clinically mild and self-limited.

Objective: To review the evidence available on the degree and duration of immunity provided by Australian tick fever vaccines against Babesia bovis, B. bigemina and Anaplasma marginale infections in Australia and overseas., Background: Vaccines containing attenuated strains of B bovis and B bigemina as well as A. centrale grown in splenectomised calves have been used in Australia since 1964 to immunise cattle against tick fever. About 800,000 doses of vaccine are supplied annually and much of the evidence for protection is field evidence rather than conventional immunological measures or pen trials., Conclusions: Immunity to Babesia bovis and B. bigemina--A single inoculation generally provides sound, long-lasting protection both in Australia and overseas. No evidence was found of a loss of immunity with time. Vaccine failures to B. bovis do occur, but are uncommon and evidently caused by a number of factors, including immune responsiveness of the vaccinated animals, and immunogenicity of the vaccine strain. Immunity to Anaplasma marginale--The vaccine containing A. centrale provides partial, variable protection against A. marginale. Protection against challenge in Australia is adequate in most cases to prevent disease and use of the vaccine in this country appears to be justified. Protection against antigenically diverse, highly virulent stocks of A. marginale in other countries is, at times, clearly inadequate and better vaccines are required in situations where the challenge is severe.

Ectoparasitic arthropods and vector-borne infectious agents are global medical and veterinary public health concerns. Economic impact due to direct effects of infestation and disease transmission are significant. These problems are increased by development of arthropod resistance to insecticides/acaricides; drug resistance of vector-borne pathogens; and, lack of effective vaccines to prevent many of these diseases. There is much to be gained from understanding the complex array of immunological interactions occurring at the arthropod-host-pathogen interface. One application of that knowledge is the development of novel vaccines for the control of both ectoparasitic arthropods and the diseases they transmit. We now realize that blood-feeding arthropods are not simply flying or crawling hypodermic needles and syringes. Ectoparasitic arthropods are not passive partners in their relationships with the immune systems of their hosts. These clever invertebrates produce numerous pharmacologically active molecules that help them migrate through tissues of their hosts or to successfully obtain blood meals. Arthropod parasites stimulate a spectrum of host immune responses that could potentially impair development, reduce feeding success, or kill the ectoparasite. Not unexpectedly, arthropods have developed sophisticated arsenals of countermeasures that modulate or deviate host immune responses. Not only does arthropod modulation of host immunity facilitate survival in tissues or increase the likelihood of obtaining a blood meal, but it is increasingly recognized as a critical factor in pathogen transmission. Those countermeasures to host immune defenses are the topics of this review. Emphasis is placed on our current understanding of the molecular bases of those changes; the molecules responsible for host immunomodulation; contemporary approaches for studying these complex relationships; and, the potential for using this information to develop innovative vaccine-based control strategies.

A cross-sectional study of the seroprevalence to Borrelia burgdorferi sensu lato and granulocytic Ehrlichia spp. in Swedish horses was conducted to evaluate associations with demographic, clinical and tick-exposure factors. From September 1997-1998, blood samples from 2018 horses were collected from the animals presented to veterinary clinics affiliated with the Swedish Horserace Totalizator Board (regardless of the primary cause for consultation). Standardized questionnaires with information both from owners and attending veterinarians accompanied each blood sample. The apparent seroprevalences to B. burgdorferi s. l. and granulocytic Ehrlichia spp. were 16.8 and 16.7%, respectively. The northern region had the lowest seroprevalences. Four logistic models were developed (controlling for demographic variables). In the disease model of seropositivity to B. burgdorferi s. l., age, breed, geographic region, the serologic titer to granulocytic Ehrlichia spp., season and the diagnosis coffin-joint arthritis were significant. In the tick-exposure model of B. burgdorferi s. l., pasture access the previous year and gender were significant. Age, racing activity, geographic region, season and the serologic titer to B. burgdorferi s. l. were associated with positivity to granulocytic Ehrlichia spp. In the tick-exposure model of granulocytic Ehrlichia spp., pasture access was a risk factor. An interaction between racing activity and geographic region showed that the risk of positive serologic reactions to Ehrlichia spp. was increased in the horse population in the south and middle of Sweden, but only among horses not used for racing. Except for the positive association between coffin-joint arthritis and serologic reactions to B. burgdorferi s. l., there were no significant associations in the multivariable models between non-specific or specific clinical sign or disease with seropositivity to either of these agents.

Theileria annulata, the cause of tropical theileriosis is propagated in cattle with stage-to-stage transmission by Hyalomma ticks. Three stages in the life cycle of the parasite--tick-derived sporozoites, intramononuclear schizonts, and erythrocytic merozoites--infect cattle. When cattle are inoculated with schizont-infected cells, the parasite is transferred from the donor cell to the recipient. The main pathological damage in cattle is induced by the schizont stage. Each development stage of T. annulata elicits a specific immune response. Schizont-infected lymphoid cells can be grown indefinitely in culture and prolonged cultivation results in loss of virulence. Blood-derived schizonts induce stronger immunity than culture-derived schizonts, which suggests that restrictions on the parasite population or antigenic variation occur during prolonged cultivation. The duration of immunity following sporozoite or schizont infections has not yet been determined, but does not appear to be lifelong. The attenuated, culture-derived anti-theileria vaccine proved to be safe and effective in prevention of field theileriosis in large enzootic areas.

In Sweden, 2 tick-borne zoonotic diseases, granulocytic ehrlichiosis and borreliosis, are frequently diagnosed in dogs, using serological assays. The aims of this study were to determine the sero-prevalences of antibodies to Ehrlichia spp. and Borrelia burgdorferi sensu lato during 1991-94 in dogs, not clinically suspected to be infected with either of the 2 agents. Samples (n = 611) were selected from a serum bank using a systematic sampling strategy, stratified across the 4-y period. The stored sera had originally been submitted in order to verify or rule out infection with Sarcoptes scabiei. The overall sero-prevalence for Ehrlichia spp. was 17.7% and for Borrelia burgdorferi sensu lato 3.9% (n = 588). Only a few dogs in the northern part of Sweden were sero-positive for Ehrlichia spp. and none were positive for Borrelia burgdorferi sensu lato. An increased sero-prevalence of Ehrlichia spp. was seen during the years studied. The sero-prevalence of Ehrlichia spp. varied with season. Sero-positivity to both agents increased with age. Both diseases are considered zoonotic, and the increase in sero-prevalence of Ehrlichia spp. over the years may reflect the degree of infection in ticks and may have implications for human health.

Anaplasmosis is widespread in South Africa with more than 99% of the total cattle population at risk. Five tick species have been experimentally shown to be capable of transmitting Anaplasma in South Africa. Mechanical transmission through blood contaminated instruments and biting flies also occurs. Vaccination against Anaplasma marginale by administration of an Anaplasma centrale live-blood vaccine has been practiced in this country since 1912. Although generally a mild pathogen, Anaplasma centrale can cause severe clinical reactions following vaccination and also does not afford complete protection against all A. marginale isolates. Anaplasmosis vaccine is routinely available in a deep-frozen form and approximately 220,000 doses of vaccine are sold per annum. Microscopic examination of stained thin blood smears is still the most reliable and cost effective method of confirming a clinical diagnosis of anaplasmosis. Several diagnostic tests, such as the complement fixation tests, card agglutination test, and enzyme-linked immunosorbent assay (ELISA) have been developed to identify carrier cattle. A competitive inhibition ELISA test, based on antibody binding to a recombinant MSP-5 protein conserved among Anaplasma species, is routinely used at this laboratory.