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Prevalence of Babesia microti in Ixodes ricinus ticks from Lublin region (eastern Poland).

A total of 1,367 Ixodes ricinus ticks collected from 5 districts of the Lublin region (eastern Poland) were examined for the presence of Babesia microti DNA by PCR and nested-PCR. As many as 74 ticks (5.4 %) were found to be infected with Babesia microti. The infection rate varied significantly with stage/sex of ticks (chi (2) =16.48543, df=2, p < 0.000264). The infection rates in females and males amounted to 6.4 % and 8.8 % respectively and were significantly higher (p=0.006 and p=0.0001 respectively) compared to minimum infection rate in nymphs that was equal to 2.8 %. The prevalence of infection showed also a significant variability depending on geographic location within the Lublin region (chi(2) =18.62812, df=4, p < 0.000932). The highest rates of infection with Babesia microti were noted in ticks collected from the areas of Puławy district situated in the northern part of region and the suburban Lublin district situated in the central part of the region (8.0 % and 7.3 % respectively). Mediocre infection rates (respectively 3.4 % and 3.3 %) were found in ticks from the Parczew and Włodawa districts situated in eastern part of the region and covered with humid lakeland forests. The lowest infection rate (0.5 %) was noted in ticks from the Zamość district situated in southern part of the region. In conclusion, the infection rate of Ixodes ricinus ticks with Babesia microti found in this study is higher compared to the majority of data reported by Polish and other European authors, and indicates a potential risk of human infection during occupational or recreational exposure to tick bite.

Animals↗

IFN-gamma IL-2, and IL-4 mRNA expression in the skin and draining lymph nodes of BALB/c mice repeatedly infested with nymphal Ixodes ricinus ticks.

The skin and draining lymph nodes of BALB/c mice were examined for IFN-gamma, IL-2, and IL-4 mRNA expression by in situ hybridization in three successive infestations with nymphal Ixodes ricinus ticks. IFN-gamma and IL-2 mRNA positive cells were readily detected in lymph node sections during primary antigenic stimulation (72 hr post-tick attachment), whereas hybridization with IL-4 probe yielded no or only a faint positive signal. No changes in the cytokine pattern were observed in lymph node sections from reinfested mice, with IL-4 mRNA always being expressed to a lesser extent than IFN-gamma and IL-2 mRNA. Seventy-two hours post-tick attachment in primary infestation, some infiltrating cells in the skin were positive for IFN-gamma and IL-4 mRNA, but not for IL-2 mRNA. In skin sections of reinfested mice, mRNA coding for IFN-gamma, IL-2, and IL-4 were detected in infiltrating cells. Cells positive for IL-4 mRNA were lower in number than those positive for IFN-gamma and IL-1 mRNA. A significant decrease in the number of IL-4 mRNA positive cells in the tertiary infestation was noted. All together, these results indicate that I. ricinus nymphal ticks antigens are able to elicit expression of IFN-gamma, IL-2 mRNA and to a lesser extent IL-4 mRNA in both skin and draining lymph nodes. In addition, repeated infestations with ticks led to strong expression of IFN-gamma and IL-2 mRNAs in the skin that may be correlated with previous observations showing the occurrence of cutaneous delayed-type hypersensitivity in tick-infested mice. Notably, the cytokine pattern observed in the skin and draining lymph nodes is not associated with a protective immune response in mice against I. ricinus nymphal ticks infestations.

Animals↗

Ticks (Acari:Ixodidae) of the Blue and White Nile ecosystems in the Sudan with particular reference to the Rhipicephalus sanguineus group.

Twenty-four adult ixodid tick species, infesting livestock and some wildlife hosts along the Blue and White Nile in the Sudan, were identified. Three species, Boophilus geigyi, Rhipicephalus camicasi and R. bergeoni, were recorded for the first time from the Sudan. Tick numbers on indigenous breeds of cattle (Bos indicus) were relatively low, ranging between 17.1 and 40.5 per animal. Young cattle grazing with the herd carried significantly fewer ticks than older animals. With the exception of B. annulatus and R. simus, which have extended their distribution further north into Blue Nile, Gezira and Khartoum Provinces, the distribution patterns of the most important cattle ticks have been relatively unchanged over the past 30 years. The Rhicicephalus sanguineus group was represented by six species. R. camicasi was the only species present on cattle, sheep and goats in the north in Kassala and Khartoum Provinces, whereas this species occurred sympatrically with R. guilhoni and R. turanicus further south in Gezira and Blue Nile Provinces. In the Southern Region of the Sudan only R. turanicus and R. guilhoni were present, the latter being by far the predominant species, with peak activity towards the end of the rains in the Jonglei Canal Area. R. bergeoni was collected once, from cattle near the Ethiopian border in Blue Nile Province, whereas R. sanguineus sensu stricto was collected throughout the study area, from domestic dogs only. Finally, R. sulcatus was found once on a hare. The distributions of the common tick species are correlated with the occurrence of tick-borne diseases of domestic animals and recommendations for the control of tick-borne diseases and their vectors in the Sudan are given.

Animals↗

Systemic activity of closantel for control of lone star ticks, Amblyomma americanum (L.), on cattle.

Cattle were treated once at 5 mg/kg orally or subcutaneously or daily at 0.1-5 mg/kg orally or 0.1-1 mg/kg subcutaneously with closantel, N-[5-chloro-4-[(4-chlorophenyl)cyanomethyl]-2-methylphenyl]-2-hydroxy-3, 5-diiodobenzamide, and numbers and weights of engorged females, weights of egg masses and hatch of eggs of lone star ticks, Amblyomma americanum, were recorded. Effectiveness of treatments on reproduction was determined by comparing total estimated larvae (EL) (EL = wt. egg mass X est. % hatch X 20000) of ticks from treated cattle with that of ticks from untreated cattle. With certain treatments, we also determined the effect of manure of treated cattle on survival of larvae of the horn fly, Haematobia irritans, or effect on survival and of fecundity of adult horn flies or stable flies, Stomoxys calcitrans, fed on blood from treated animals. The single oral treatment afforded essentially complete control of total EL only of ticks placed on the animal on the day of treatment, while the single subcutaneous treatment afforded greater than 92% control of total EL of ticks placed on animal on treatment day and for 6 weeks posttreatment. Daily treatments of 0.5 mg/kg or greater orally and 0.1 mg/kg or greater subcutaneously afforded essentially complete control of total EL of ticks throughout the treatment period (3-12 weeks) and for 1-7 weeks after treatment was discontinued. An estimated concentration of greater than 9 micrograms/ml of blood was calculated by probit analysis to be necessary to provide greater than 90% control of total EL of lone star ticks; that same concentration also provided greater than 90% control of hatch of eggs laid by treated females. A higher concentration (40 micrograms/ml) was necessary to prevent engorging of the females. No treatments tested were effective against larvae of the horn fly or adult horn flies or stable flies.

Animals↗

Control of induced infestations of three African multihost tick species with sustained-release ivermectin.

The efficacy of ivermectin, released intraruminally from a 28-day-delivery device was evaluated in two titration studies against induced infestations of adult Rhipicephalus appendiculatus, R. evertsi and Hyalomma truncatum on cattle. Cattle were given a sufficient number of devices to release ivermectin at approximately 20, 40, 60 or 80 micrograms kg-1 day-1 at a steady-state rate 7-28 days after administration. Tick mortality was recorded, engorged female ticks were weighed and individually incubated, and reproductive data were recorded to determine a reproductive index for the species at various dose levels. Mortality of male and female ticks compared to that of controls was directly related to the daily dose of ivermectin, as was the number of ticks not engorging. Ticks fed on ivermectin-treated cattle had a smaller mass when engorged and laid smaller egg-masses, both absolutely and as a proportion of engorged mass. The index of reproduction of R. appendiculatus was reduced by more than 99.9% at 20 micrograms kg-1 day-1, and the reproductive indices of R. evertsi and H. truncatum were reduced by more than 99.9% at dose rates of 40 micrograms kg-1 day-1 and above. Practical implications of the application of sustained-release ivermectin for the control of multihost ticks and tick-borne diseases are discussed.

Animals↗

Two-year survey of the incidence of Lyme borreliosis and tick-borne encephalitis in a high-risk population in Sweden.

A survey was made over a two-year period (September 1987 to August 1989) of a population living in an area endemic for Lyme borreliosis and tick-borne encephalitis in Sweden. For each patient a blood sample was collected and a questionnaire completed annually. All sera were tested for an antibody response to Borrelia burgdorferi in an EIA using sonicated antigen and for an antibody response to the tick-borne encephalitis virus using an EIA and a haemagglutination inhibition test. Antibodies to Borrelia burgdorferi and tick-borne encephalitis virus were detected in 89 (25.7%) and 40 (11.6%) respectively of 346 samples collected in August 1987. In the first year of the study 14 of 303 subjects (4.6%) developed Lyme borreliosis and in the second year 9 of 277 subjects (3.2%). A significant increase in the antibody titre for Borrelia burgdorferi was seen in 14 of 303 (4.6%) subjects in the first year and 8 of 277 (2.9%) subjects in the second year. An earlier episode of Lyme borreliosis or an elevated antibody titre did not seem to protect against reinfection. One case of tick-borne encephalitis was seen each year. Seroconversion for tick-borne encephalitis virus was found in 3 of 258 (1.2%) subjects in the first year and 5 of 211 (2.4%) in the second year, excluding subjects who had undergone successful immunisation or had earlier been hospitalised for tick-borne encephalitis. The study thus demonstrated a high yearly incidence of tick-borne infections in a population at risk.

Adolescent↗

Amblyomma americanum: requirement for host Fc receptors in antibody-mediated acquired immune resistance to ticks.

Guinea pig recipients of anti-tick immune serum or immune peritoneal exudate cells expressed 25 and 30% tick rejection, respectively, when challenged with Amblyomma americanum larval ticks. Previous studies have shown that IgG1 antibodies are responsible for the ability of immune serum to transfer resistance to ticks and to mediate the accompanying, and required, cutaneous basophil response. Since IgG1 antibodies induce mast cell-mediated passive cutaneous anaphylaxis and cutaneous basophil responses by interaction with cell surface Fc receptors, we investigated whether host Fc receptors were involved in the mechanism of antibody-mediated immune resistance to ticks. Recipients of immune serum pretreated intravenously with rabbit IgG failed to express resistance when challenged. In contrast, recipients of immune peritoneal exudate cells similarly pretreated expressed normal resistance. Sheep IgG had no inhibitory effect on the transfer of resistance by either immune serum or peritoneal exudate cells. Furthermore, recipients of immune serum pretreated with the Fc fragment from papain digestion of rabbit IgG failed to express resistance when challenged with ticks. Rabbit Fab and sheep Fc and Fab had no effect on the transfer of resistance by immune serum. Purity of rabbit Fc preparations was verified by the ability to inhibit mast cell-mediated passive cutaneous anaphylaxis due to high-titered IgG1 antiovalbumin antibodies. Rabbit Fab and sheep Fc fractions did not inhibit passive cutaneous anaphylaxis reactions. These findings suggest that immunoglobulin Fc receptors on host cells, such as mast cells and basophils, are required for antibody-mediated immune rejection of ticks from guinea pigs.

Animals↗

Loss of Lyme disease spirochetes from Ixodes ricinus ticks feeding on European blackbirds.

To determine whether blackbirds (Turdus merula), the most abundant and most abundantly tick-infested ecotonal bird of Central Europe, may contribute to the transmission of the Lyme disease spirochete (Borrelia burgdorferi), we compared the infectivity to ticks of naturally as well as experimentally infected blackbirds and rodents. European blackbirds experience intense exposure to Ixodes ricinus ticks and to the pathogens that they transmit. In nature, subadult I. ricinus ticks found feeding on these birds generally contain no spirochetes, although infection is universal in those found on black-striped mice (Apodemus agrarius). Those found on yellow-necked mice (A. flavicollis) are less frequently infected. Ticks lose infection in the course of feeding on blackbirds and fail to infect them. Subadult I. ricinus ticks readily feed on blackbirds, black-striped mice, and jirds (Meriones unguiculatus), but engorge less fully on the bird than on the rodents. Although birds may burden human health by establishing new infestations of I. ricinus ticks, our observations indicate that particular birds may benefit health by locally diminishing transmission of the Lyme disease spirochete.

Animals↗

Efficacy of ivermectin delivered from an intraruminal sustained-release bolus against natural infestations of five African tick species on cattle.

The efficacy of ivermectin delivered by an orally administered prototype sustained-release bolus over approximately 90 days was evaluated against natural infestations of five African tick species. Twenty cattle, allocated by restricted randomization based on counts of standard Boophilus decoloratus, were allocated to two groups and were either given an ivermectin bolus or designated as non-medicated controls. All cattle grazed a single pasture of native grasses for 20-40 days before treatment and until trial termination. Starting on Days 27, 40, 68 and 82 after bolus administration, four replicates were confined to individual tick-collection stanchions for 4 to 5-day periods. Ticks recovered from these cattle were counted by species, sex, and stage and degree of repletion; engorged females were weighed and incubated to determine the number which oviposited. For the other replicates, half-body counts of adult ticks (classified by species, sex and degree of repletion by females) were made at 1- and 2-week intervals through Day 90. Among replicates confined to stanchions periodically, fewer (P less than 0.05) engorged adult female B. decoloratus, Hyalomma spp., Rhipicephalus appendiculatus and Rhipicephalus evertsi evertsi were collected from bolus-treated cattle than from controls. Numbers of engorged adult female Amblyomma hebraeum were reduced, but differences were not statistically significant (P greater than 0.10). Among cattle maintained continuously on pasture, tick numbers were reduced on the ivermectin-treated groups. A significant (P less than 0.05) treatment by linear time effect was seen for all adult ticks counted except R. appendiculatus. A significant (P less than 0.05) treatment by quadratic time effect was seen for A. hebraeum, B. decoloratus and R. evertsi evertsi, and overall treatment differences were significantly different (P less than 0.05) for these species. The differences tended to increase with time. Except for Boophilus, reductions in tick numbers on treated animals relative to controls were not readily apparent. There were no adverse reactions attributable to ivermectin treatment or the presence of the bolus. Each treated animal retained its bolus throughout the trial, based on metal detection.

Administration, Oral↗

Histopathology of attachment sites of Amblyomma variegatum and Rhipicephalus appendiculatus on zebu cattle of varying resistance to ticks.

Mature, unimproved East African Zebu cattle (Bos indicus) naturally exposed to ticks from birth were ranked for resistance to ticks by repeated whole-body counts of infesting ticks. Four cattle of high resistance, four of low resistance and two of intermediate resistance were artificially infested with nymphs of Amblyomma variegatum and Rhipicephalus appendiculatus. Two Friesian cattle (Bos taurus) with less exposure to ticks were similarly infested. Biopsies of tick attachment sites were examined histologically. All attachment sites showed acute inflammatory lesions, and sites of both tick species on high resistance cattle showed delayed hypersensitive reactions with intra-epidermal pustulation and significant increases in the numbers of granulocytes. The predominant cells infiltrating attachment sites on high resistance cattle were eosinophils with A. variegatum and neutrophils with R. appendiculatus. Such differences need to be taken into account in developing immunological tests for selecting cattle for resistance but there are sufficient common features of reactions to the two infesting tick species to justify dermal hypersensitivity tests.

Africa, Eastern↗

Laboratory and field observations on anti-tick properties of the plant Gynandropsis gynandra (L.) Brig.

A shrubby plant, abundant in east Kenya, Gynandropsis gynandra (L.) Brig., was shown to exhibit repellent and acaricidal properties to larvae, nymphs and adult Rhipicephalus appendiculatus and Amblyomma variegatum ticks. All stages of ticks avoided the leaves of the plant and a high percentage of the ticks which were continuously exposed to its leaves died; surviving ticks were weak and inactive but regained activity when exposed to fresh air. The effectiveness of the plant's leaves as a repellent and acaricide was most pronounced on nymphs and least pronounced on adults. Field investigations indicated that ticks were not found up to 2-5 m from the plant in areas where the plant was predominant. The potential of using the plant for tick control within an integrated tick management system in the resource-poor farming context in Africa was highlighted.

Animals↗

Relationships and influences between Boophilus microplus characteristics in tick-naive or repeatedly infested cattle.

Six tick-naive male Hereford calves were infested once a month for 6 months with 18,000 Boophilus microplus larvae on the back and with 400 larvae in a cloth bag glued on the lumbar region. Working with the bag ticks, 12 tick characteristics were recorded for each infestation. Each tick attribute was analyzed for significant differences with those of the first infestation (analysis of variance), and for similarity (clustering), degree of relationship (correlation), and concomitant variation (regression) against all the other attributes during the first, third, and sixth infestations. Some attributes were affected maximally by host immunity about the third infestation but recovered later (length of feeding, detachment weight, egg weight, start of oviposition, fertility efficiency index), whereas others continued to be affected until the last infestation (length of oviposition, corpse weight, start of hatching, feeding efficiency index). All analyses showed that weight at detachment and egg weight were closely related, and corpse weight was partially related to these two. All other natural characteristics were largely independent. Length of feeding showed no significant relation with weight at detachment nor length of oviposition with egg weight. These findings suggest that different tick functions are independently affected by host immunity and recommends against estimating general anti-tick resistance by the evaluation of only a few tick characteristics.

Analysis of Variance↗

Threats to international travellers posed by tick-borne diseases.

To date, 14 tick-borne diseases have been reported in international travellers, the majority of cases being Lyme borreliosis caused by Borrelia burgdorferi sensu lato in North America and Eurasia, African tick bite fever caused by Rickettsia africae in sub-Saharan Africa and eastern Caribbean, and Central European encephalitis caused by tick-borne encephalitis virus in Europe. The clinical presentation is frequently non-specific, and tick-borne diseases should always, in the absence of other likely diagnoses, be suspected in travellers with flu-like symptoms following a recent visit to tick-infested areas. Feasible microbiological diagnostic tests are widely unavailable, at least outside areas of endemicity where many infected travellers present. Empiric treatment with doxycycline should be considered in suspected cases of tick-borne bacterial diseases. Since ecotourism and adventure travel are increasingly popular worldwide, the incidence of travel-associated tick-borne diseases is likely to increase in the future.

Animals↗

Two serine protease inhibitors (serpins) that induce a bovine protective immune response against Rhipicephalus appendiculatus ticks.

We have previously undertaken preliminary characterization of two Rhipicephalus appendiculutus serine protease inhibitors (RAS-1 and -2) as anti-tick vaccine candidates. In this study, to clarify this hypothesis, we generated and further characterized recombinant RAS-1 and -2 (rRAS-1 and -2) and tested their potency as a cocktail anti-tick vaccine in cattle. RT-PCR analysis showed that RAS-1 and -2 mRNA transcripts are expressed during all life cycle stages of ticks, independent of sex. As judged by SDS-PAGE rRAS-1 and -2 migrated as a molecular weight of around 64 and 60 kDa protein, respectively, considering that the expression vector produced a recombinant protein fused with 18-22 kDa TRX protein. RAS-1 and -2 were found not to be secreted into the bite site as determined by the reactivity of anti-tick saliva sera to rRAS-1 and -2, suggesting that both proteins are concealed antigens. Vaccination of cattle with a combination of rRAS-1 and -2 conferred significant protective immunity against ticks, resulting in 61.4% reduction in nymph engorgement rate, and in 28 and 43% increased mortality rate in adult female and male ticks, respectively. This is the first report on an anti-tick vaccine trial using a combination of two different serpins derived from R. appendiculatus, and using cattle as a natural host.

Animals↗

A cement protein of the tick Rhipicephalus appendiculatus, located in the secretory e cell granules of the type III salivary gland acini, induces strong antibody responses in cattle.

Protein components of the cement cone of ixodid ticks are candidates for inclusion in vaccines against tick infestation, since they are essential for tick attachment and feeding. We describe here the cloning of a cDNA encoding a 36 kDa protein, designated Rhipicephalus Immuno-dominant Molecule 36 (RIM36), present in salivary glands and the cement cone material secreted by Rhipicephalus appendiculatus. The 334-amino-acid sequence of RIM36 has a high content of glycine, serine and proline. The protein contains a predicted N-terminal signal peptide and two classes of glycine-rich amino acid repeats, a GL[G/Y/S/F/L] tripeptide and a GSPLSGF septapeptide. Comparison of genomic and cDNA sequences reveals a 597 bp intron within the 3' end of the RIM36 gene. Immuno-electron microscopy demonstrates that RIM36 is predominantly located in the e cell granules of the type III salivary gland acini. An Escherichia coli recombinant form of the proline-rich C-terminal domain of RIM36 reacts with antisera from Bos indicus cattle, either experimentally infested with R. appendiculatus, or exposed to ticks in the field. The 36 kDa protein is strongly recognised on Western blots of salivary gland lysates and soluble extracts of purified R. appendiculatus cement cones by polyclonal antibodies generated against recombinant RIM36, and by antisera from cattle experimentally infested with ticks. The data indicate that this tick cement component is a target of strong antibody responses in cattle exposed to feeding ticks.

Amino Acid Sequence↗

Tick-borne encephalitis in Sweden and climate change.

BACKGROUND: The incidence of tick-borne encephalitis (TBE) in Sweden has substantially increased since the mid-1980s. During the same period the climate has become milder and ticks have become more abundant. We investigated whether there is a link between the change in climate and the increase in incidence of TBE. METHODS: Since the late 1950s all cases of encephalitis admitted in Stockholm County have been serologically tested for TBE. We analysed the period 1960-98 with multiple regressions. The number of days per season with temperatures of known importance for tick prevalence and pathogen transmission were studied. 2 years of temperature data were related to each TBE incidence rate to account for the tick's long life-span. FINDINGS: Increases in disease incidence was significantly related (R(2)=0.58; p<0.0001) to a combination of two consecutive mild winters, temperatures favouring spring development (8-10 degrees C) and extended autumn activity (5-8 degrees C) in the year prior to the incidence year, and temperatures allowing tick activity (5-8 degrees C) early in the incidence year. INTERPRETATIONS: The findings indicate that the increase in TBE incidence since the mid-1980s is related to the period's change towards milder winters and early arrival of spring. Other factors may have influenced TBE incidence such as more people in endemic locations, and increases in host animal populations; factors which are partly climate related. Access to TBE vaccination since 1986 and increased awareness of ticks might have caused an underestimation of the links found. Our findings also suggest that the incidence of other tick-borne zoonoses might have been affected by the milder climate.

Animals↗

Effect of controlling natural field-tick infestation on the growth of N'Dama and Gobra zebu cattle in the Gambia.

The effect of tick infestations on liveweight gain (LWG) was assessed by comparison of weight changes in flumethrin-treated N'Dama and Gobra zebu cattle (16-20 months old) with respective control groups submitted to natural tick challenge over 1 year. Flumethrin was applied monthly, fortnightly or weekly. Preventive treatments against anaplasmosis, babesiosis and trypanosomosis were given. Mortality rate was recorded and post-mortem examinations carried out. In both treated and control animals, significantly fewer Hyalomma spp. and Amblyomma variegatum were found on N'Dama than on Gobra zebu cattle. Both breeds are equally susceptible to Rhipicephalus senegalensis infestation. Total annual tick burdens did not cause significant differences in LWG between acaricide-treated and control cattle in either breeds. LWG was also not affected during or after the annual peak of tick infestation (composed mainly by A. variegatum and R. senegalensis). Equally-high mortality (35%), due to unidentified causes, was recorded in acaricide-treated and control Gobra cattle; mortality in N'Dama cattle was 7.5%. In both breeds, about the 90% of mortality occurred at the end of the dry season. Breed differences in tick burden confirm previous results. If tick-borne infections do not influence LWG or mortality, then it is concluded that intensive tick control is not justifiable in Gambian livestock.

Anaplasmosis↗

Field studies and cost-effectiveness analysis of vaccination with Gavac against the cattle tick Boophilus microplus.

The control of tick infestations and the transmission of tick-borne diseases remains a challenge for the cattle industry in tropical and subtropical areas of the world. Traditional control methods have been only partially successful and the parasites continue to result in significant losses for the cattle industry. Recently, vaccines containing the recombinant Boophilus microplus gut antigen Bm86 have been developed. These vaccines have been shown to control tick infestations in the field. However, extensive field studies investigating the efficacy and cost-effectiveness of vaccination have not been reported and are needed to appraise the effect of this new approach for tick control. Here is reported the results of the application of Gavac in a field trial including more than 260,000 animals in Cuba. In this study the correlation between the antibody response to vaccination and the effect on ticks fertility is determined. Physiological status of the animals was found to affect the primary response to vaccination but not the antibody titers after revaccination. A cost-effectiveness analysis showed a 60% reduction in the number acaricide treatments, together with the control of tick infestations and transmission of babesiosis, which resulted in savings of $23.4 animal-1 year-1. These results clearly demonstrate the advantage of vaccination and support the application of Gavac for tick control.

Animals↗