Comparison of known and suspected pheromonal constituents in males of the African ticks,Amblyomma hebraeum Koch andAmblyomma variegatum (Fabricius)

Three low molecular weight compounds were found in hexane: diethyl ether extracts of fed males of the African ticks,Amblyomma variegatum (tropical bont tick) andA. hebraeum (bont tick), namely,o-nitrophenol, methyl salicylate and 2,6-dichlorophenol. These same compounds were also found in a rinse of fedA. variegatum males, but were absent or present in only trace amounts in a rinse of fedA. hebraeum males.o-Nitrophenol and methyl salicylate were present in much higher concentrations (i.e., amounts/tick) inA. variegatum than inA. hebraeum. 2,6-Dichlorophenol was also more abundant inA. variegatum than inA. hebraeum, but the differences were not as great as with the former two compounds. Extraction in hexane over a 3-week period revealed four additional compounds, benzaldehyde, benzyl alcohol, benzothiazole and nonanoic acid. The first three compounds were found in males of both species; nonanoic acid was found only inA. hebraeum males. Published reports consistently show strong attraction byo-nitrophenol and methyl salicylate for both sexes of the two bont tick species; 2,6-dichlorophenol and benzaldehyde have been reported to be attractive to both sexes ofA. hebraeum. The possible roles of these compounds, as well as others occasionally reported fromA. hebraeum andA. variegatum, as components of the aggregation/attachment pheromone or other pheromones is discussed.Supported by Cooperative Agreement No. AFR-0435A-00-9084-00 with the U.S. Agency for International Development to the Department of Infectious Diseases, College of Veterinary Medicine, University of Florida, Gainesville, FL.     

Efficacy of tags impregnated with pheromone and acaricide for control of Amblyomma variegatum

The efficacy of tags impregnated with pheromone and acaricide for control of Amblyomma variegatum on cattle in Guadeloupe was determined for a 13-week trial. Comparisons were made between untreated cattle and cattle with tags containing either pheromones alone (o-nitrophenol, methyl salicylate, 2,6-dichlorophenol and phenylacetaldehyde), pheromones plus acaricide (cyfluthrin or deltamethrin), or acaricide alone. Tags were fastened to cattle both on collars on the neck and with adhesive to tail hairs, with over 98 and 90% of tags retained, respectively, during the trial. By the end of the trial, tick infestations increased on untreated cattle (311.7%) and cattle with pheromone tags (154.8%) but decreased on cattle with pheromone/cyfluthrin (–45.0%), cyfluthrin (–42.8%), pheromone/deltamethrin (–68.7%) and deltamethrin tags (–87.6%). Cattle with pheromone tags had greater proportions of ticks on the hind regions (81%) compared to untreated cattle (62.5%) and on the front regions (18.2%) compared to untreated cattle (8.2%) indicating that ticks aggregated in response to pheromones. Analysis of hair samples by gas chromatography indicated that, within 2 days of placement, cyfluthrin spread rapidly over the body from the tags. Both cyfluthrin and deltamethrin were present in detectable levels on all eight body regions examined throughout the trial. Analysis of the tags indicated that they all still contained pheromone components after 13 weeks under field conditions.     

Responses of the ticksAmblyomma hebraeum andA. variegatum to known or potential components of the aggregation-attachment pheromone. III. Aggregation

Ten known or potential components of the aggregation-attachment pheromone (AAP) of the ticksAmblyomma hebraeum andA. variegatum, as well as mixtures of these components, extracts of prefed males and live prefed males, were tested as aggregation stimulants. In field assays, laboratory-reared unfed male and female ticks were released 20 cm downwind of CO₂/pheromone release sites; the numbers of ticks that aggregated at the release sites were recorded after 30 min. InA. variegatum, aggregation was induced by methyl salicylate,o-nitrophenol, 2,6-dichlorophenol, phenylacetaldehyde and some mixtures containing these compounds; a strong aggregation response was induced by an extract of five prefed malesA. variegatum and a weak response was induced by an extract of 50 prefed males ofA. hebraeum. InA. hebraeum, aggregation was induced by phenylacetaldehyde, mixtures of compounds that included phenylacetaldehyde, extracts of 50 prefed males ofA. hebraeum orA. variegatum and 50 live prefed males ofA. hebraeum. InA. variegatum, aggregation was inhibited if compounds that do not occur naturally in the AAP of the species were included in mixtures. InA. hebraeum, phenylacetaldehyde appeared to act as an arrestant for ticks that had been attracted to release sites by other compounds.     

Responses of the ticksAmblyomma hebraeum andA. variegatum to known or potential components of the aggregation-attachment pheromone. I. Long-range attraction

Nine known or potential components of the aggregation-attachment pheromone of the ticksAmblyomma hebraeum andA. variegatum were tested as long-range attractants. In field assays, laboratory-reared unfed male and female ticks were released 4 m downwind of treated filter-paper targets. Carbon dioxide was used to activate the ticks and the numbers attracted to the targets were recorded. Adults ofA. hebraeum were attracted to six compounds (benzaldehyde, 2,6-dichlorophenol, heptadecane, methyl salicylate, nonanoic acid and o-nitrophenol) and adults ofA. variegatum to four compounds (2,6-dichlorophenol, methyl salicylate, nonanoic acid and o-nitrophenol). The most important attractant forA. hebraeum was o-nitrophenol; methyl salicylate and o-nitrophenol were the most important attractants forA. variegatum. Nonanoic acid and o-nitrophenol attracted significantly more female than maleA. hebraeum. Both species were also attracted by a mixture of the nine compounds.     

Responses of the ticksAmblyomma hebraeum andA. variegatum to known or potential components of the aggregation-attachment pheromone. IV. Attachment stimulation of nymphs

Ten known or potential components of the aggregation-attachment pheromone (AAP) of the ticksAmblyomma hebraeum andA. variegatum, as well as a mixture of these components and extracts of prefed males of the two species, were tested as attachment stimulants for nymphs. Unfed nymphs were confined in linen bags on the ears of rabbits that had been treated with the test compounds, mixture or extracts; the numbers attached were recorded after 24 h. InA. hebraeum, attachment was induced by four compounds (2-methyl propanoic acid, methyl salicylate,o-nitrophenol and salicylaldehyde), the mixture and extracts from both species. InA. variegatum, attachment was induced by three compounds (methyl salicylate,o-nitrophenol and salicylaldehyde), the mixture and extracts from both species. Methyl salicylate ando-nitrophenol are primary components of the AAP of bothA. hebraeum andA. variegatum. 2-methyl propanoic acid is a species-specific attachment stimulant forA. hebraeum. Salicylaldehyde, a phenolic compound, is not a naturally occurring AAP component. Nymphs of both species respond to fewer attachment stimulants than the adults and, as shown by their respective host ranges, are less dependent on the AAP in the regulation of attachment than the adults.     

Attraction of Amblyomma variegatum (ticks) to the attraction-aggregation-attachment-pheromone with or without carbon dioxide

The responses of adult Amblyomma variegatum ticks released from various distances to different doses of the synthetic attraction-aggregation-attachment pheromone (AAAP) (made up of ortho-nitrophenol, methyl salicylate and nonanoic acid in paraffin oil), dispensed from the center of circular field plots, were studied in the presence or absence of elevated levels of CO₂. Up to 90% of the ticks released were attracted to the pheromone source in the presence of CO₂ within 3h. CO₂ alone was unattractive, similar to previous findings in Zimbabwe, but unlike results from a Caribbean A. variegatum population, which was significantly attracted to this signal. In the absence of CO₂, smaller but significant proportions of the released ticks were attracted to the pheromone, albeit more slowly, suggesting another variation in the responses of this bont tick to inter- and intra-specific signals. Our results are interpreted in the light of a study undertaken elsewhere demonstrating relatively high heterozygosity among tick populations. Possible directions of further research to explore the use of the pheromone in off-host control of the tick are also highlighted.     

The aggregation-attachment pheromone of the tropical bont tick Amblyomma variegatum Fabricius (Acari,Ixodidae): Isolation,identification and action of its components

Combining a new in vitro bioassay with the analytical method capillary gas chromatography-mass spectrometry, the aggregation-attachment pheromone produced by fed males of the tropical bont tick Amblyomma variegatum was shown to consist of o-nitrophenol, methyl salicylate and pelargonic acid in the approximate amounts of 2/1/8 × 10^?6 g/tick. A synthetic pheromone blend composed of those three volatile compounds evoked an aggregation response in unfed males and females in a bioassay comparable to the response to a natural pheromone source. Of the individual components. only o-nitrophenol induced a significant, although not complete aggregation response. Methyl salicylate and pelargonic acid contribute to complete pheromone activity, but induce no aggregation response at all, when offered separately.     

Specificity and effects of host resistance on the African tickRhipicephalus zambeziensis (Acarina: Ixodidae)

Repeated infestations of the rabbit with adultRhipicephalus zambeziensis ticks resulted in a tenfold reduction in engorgement weight of adult ticks and a significant decline in engorgement weight of nymphs. There was no evidence of cross-resistance against the South African bont tick,Amblyomma hebraeum. Immunization of rabbits with crude extracts of whole-tick protein significantly influenced engorgement weights and oviposition rates of adultR. zambeziensis.Histological studies of tissues fromR. zambeziensis adults fed on resistant rabbits showed severe gut damage; this was associated with the binding of host IgG to mid-gut cells.     

Use of carbon dioxide and natural or synthetic aggregation-attachment pheromone of the bont tick,Amblyomma hebraeum,to attract and trap unfed adults in the field

It was determined previously that unfed adults ofAmblyomma hebraeum become active in response to carbon dioxide but are not attracted by it. We have now shown in field experiments that males and females which are activated by carbon dioxide are attracted to the aggregation-attachment pheromone produced by males after a period of feeding on the host. Unfed adults were attracted too-nitrophenol, a component of the aggregation-attachment pheromone ofAmblyomma variegatum, pre-fed males ofA. hebraeum, and extracts of such males. Live males were more attractive than the extracts which, in turn, were more attractive thano-nitrophenol. The ticks were attracted to sources ofo-nitrophenol from as far away as 11 m in 90 min. Simple traps, baited with carbon dioxide and aggregation-attachment pheromone (from pre-fed males or extracts of pre-fed males), were used to capture unfed adults that had been released in the field. The response of unfed adults to carbon dioxide and the aggregation-attachment pheromone presents an obvious host-location mechanism and a means for the ticks to discriminate between suitable hosts (those with males already attached) and potentially unsuitable hosts (those with no males attached).     

Responses of the ticksAmblyomma hebraeum andA. variegatum to known or potential components of the aggregation-attachment pheromone. II. Attachment stimulation

Nine known or potential components of the aggregation-attachment pheromone of the ticksAmblyomma hebraeum andA. variegatum were tested as attachment stimulants. Unfed adult male and female ticks were confined in linen bags on the ears of rabbits that had been treated with solutions of the test compounds; the numbers attached were recorded after 24 h. In both species, males attached more rapidly and more readily than females. InA. hebraeum, attachment of males was induced by eight compounds (benzaldehyde, benzyl alcohol, 2,6-dichlorophenol, heptadecane, 2-methyl propanoic acid, methyl salicylate, o-nitrophenol and salicylaldehyde) and attachment of females by six compounds (benzaldehyde, benzyl alcohol, 2,6-dichlorophenol, 2-methyl propanoic acid, o-nitrophenol and salicylaldehyde). InA. variegatum, attachment of males was induced by seven compounds (benzaldehyde, benzyl alcohol, 2,6-dichlorophenol, heptadecane, methyl salicylate, o-nitrophenol and salicyladeehyde) and attachment of females by two compounds (methyl salicylate and o-nitrophenol). A mixture of the nine compounds and extracts of prefed males ofA. herbraeum andA. variegatum induced attachment in males of both species and in femaleA. hebraeum; in these tests, attachment of femaleA. variegatum was only induced by extracts of conspecific males.     

Resistance of Brahman and Hereford Cattle to African ticks with reference to serum gamma globulin levels and blood composition

Field collections of ticks from two breeds of cattle showed that the common species of ticks wereAmblyomma hebraeum, Rhipicephalus appendiculatus andRhipicephalus evertsi evertsi. The density of these species was higher on Hereford than on Brahman cattle. The results also indicated that the density of immature stages of the above-mentioned ticks is higher on Hereford than on Brahman cattle.A positive correlation was found between the number of ticks on the cattle and the serum gamma globulin levels, an indication of an increase in the production of antibodies. An inverse relationship was found between tick burden and red blood cell count and hemoglobin concentration. Other ablood components, such as basophils, eosinophils and lymphocytes were not affected by the changes in the density of the tick populations. It appears that resistance may have been acquired by the hosts and that Brahman cattle may acquire resistance to a higher degree than Herefords.     

Responses of the tropical bont tick, Amblyomma variegatum (Fabricius), to its aggregation-attachment pheromone presented in an air stream on a servosphere

Male Amblyomma variegatum ticks feeding on a host release a mixture of o -nitrophenol and methyl salicylate which serves to attract conspecifics. The behavioural responses of A. variegatum on a servosphere to these volatiles presented in an air stream are detailed here. In still air, ticks walked on all eight legs, but with long halts. In contrast, the air stream caused continuous walking and induced a reaching response where the forelegs actively sampled the air. Such reaching increased the angular velocity and reduced walking speed, effects that were amplified in the presence of vapours from o -nitrophenol and methyl salicylate in the air flowing over the ticks. Vapour from a 1:1 mixture of o -nitrophenol and methyl salicylate was attractive over a 10⁴-fold concentration range providing an increase in upwind displacement of 20–40%, significantly higher than the natural ratio where o -nitrophenol vapour predominates. Although the responses to o -nitrophenol vapour were variable when presented alone, this chemical was consistently attractive when delivered with steer hair odour – unattractive on its own. Moreover, the upwind walk to this combination did not cause a change in speed or angular velocity. This supports the hypothesis that the response to the pheromone is enhanced by host odour. Accepted: 16 October 1999     

An update on the ecological distribution of Ixodid ticks infesting cattle in Rwanda: countrywide cross-sectional survey in the wet and the dry season

As part of the epidemiological studies aimed at developing an East Coast fever (ECF) immunisation control strategy, which combines an infection and treatment method with strategic tick control, a countrywide tick survey was carried out in both the dry and the wet season to determine the abundance and the dynamics of the tick populations infesting cattle in Rwanda. Six Ixodid tick species where identified from a total of 12,814 tick specimens collected. Rhipicephalus appendiculatus, the main vector of ECF was the most abundant (91.8%) followed by Boophilus decoloratus (6.1%) and Ambyomma variegatum (1.2%). Few ticks from the three other less economically important Ixodid species (Rhipicephalus compositus, R. evertsi evertsi and Ixodes cavipalpus) were recovered. Both adult and immature stages of the most dominant tick species were found to be widespread with a year round presence. The numbers of ticks were high in low land and medium zones and declined markedly in the higher regions of Rwanda. The geographical distribution of various tick species throughout the country and their epidemiological implications are discussed.     

Pheromonal composition of two species of african Amblyomma ticks: similarities,differences and possible species specific components

Two species of bont ticks, Amblyomma hebraeum and Amblyomma variegatum, common to Africa were studied to compare types and quantities of compounds known or believed to serve as components of the attraction-aggregation-attachment pheromone (AAAP). A complex of attraction, aggregation and attachment stimulating pheromone components are used by these to detect hosts, mates and, perhaps, minimize interspecific breeding. Solvent extraction of pheromone emitting ticks followed by gas chromatography and mass spectrometry revealed little qualitative difference in the composition of the AAAP in these two species. However, subtle differences in the relative proportions of the pheromonal blend are noted suggesting that such differences in concentration may facilitate species-specific discrimination during aggregation and attachment. Differences in the relative abundance of benzaldchyde and methyl salicylate in the males of the two species were especially noteworthy. Possible means by which such differences in phenolic compound composition may affect the behavior of these ticks are discussed.     

Factors affecting the distributions of the ticks Amblyomma hebraeum and A. variegatum in Zimbabwe: implications of reduced acaricide usage

The ticks Amblyomma hebraeum and A. variegatum are the main vectors of heartwater, a disease of ruminants caused by Cowdria ruminantium, in the agricultural areas of Zimbabwe. At present, A. hebraeum is widely distributed in the dry southern lowveld, and occurs in at least seven foci in the higher rainfall highveld. Amblyomma variegatum occurs in the Zambezi valley and surrounding dry lowveld areas in the northwest. The distribution of A. hebraeum has changed considerably over the past 70 years, while that of A. variegatum appears to have remained fairly static. The distribution patterns of both species in Zimbabwe display anomalous features; the ticks occur in areas of lowest predicted climatic suitability for survival and development and in areas where the densities of cattle, the most important domestic host, are lowest. The only factor favouring the survival of the species in the lowveld habitats in which they occur is the presence of alternative wildlife hosts for the adult stage. Their absence from more climatically favourable highveld habitats appears to have been the result of intensive acaricide treatment of cattle over a long period and a historic absence of significant numbers of wildlife hosts. Eradication of A. hebraeum and A. variegatum by intensive acaricide treatment of cattle can be achieved in the absence of significant numbers of alternative hosts, because of the long attachment and feeding periods of the adults of these tick species. However, eradication becomes impossible when alternative hosts for the adult stage are present, because a pheromone emitted by attached males attracts the unfed nymphal and adult stages to infested hosts. The unfed ticks are not attracted to uninfested hosts, such as acaricide-treated cattle.Regular acaricide treatment of cattle is expensive and so, for economic reasons, the Government of Zimbabwe is no longer enforcing a policy of strict tick control. It is likely that reduced tick control will result in the spread of Amblyomma ticks to previously uninfested areas. Added to this, recent introductions of various wildlife species to highveld commercial farming areas have created conditions in which the ticks could become established in higher rainfall areas. Amblyomma hebraeum is more likely to spread than A. variegatum, because its adults parasitize a wider range of wildlife hosts (warthogs, medium to large-sized antelope, giraffe, buffalo and rhinoceros), whereas adults of A. variegatum appear to be largely restricted to one wildlife species (buffalo) in Zimbabwe, the distribution of which is now confined to very limited areas of the country, as part of foot and mouth disease control measures. A model to predict the rate of spread of A. hebraeum through the highveld is described.Possible control options for dealing with the spread of Amblyomma ticks and heartwater to previous unaffected highveld areas, include (1) continuation of intensive acaricide treatment of cattle to prevent the spread, (2) establishment of a buffer zone of intensive tick control around affected areas to contain the spread and (3) allow the spread to occur and control heartwater by means of immunization. An economic analysis to determine the costs and benefits of the control options, which takes into account the development of Amblyomma-specific tick control technologies and improved heartwater vaccines, is recommended.Deceased.     

Morphological identification of ticks (Acari: Ixodidae) infesting donkeys (Equus asinus) in Maputo Province,Mozambique

Donkeys (Equus asinus) are rustic animals, but in Africa’s poorest regions, they can present multiple health problems, including tick infestation. The study’s objective was to determine the species composition of ticks that infest donkeys in Maputo Province (Mozambique). Ticks were collected in five conveniently selected southern districts of Maputo Province (Moamba, Matutuíne, Marracuene, Boane, and Matola) and were identified to species level using a stereoscopic microscope with the aid of dichotomous identification keys. In total, 500 ticks were collected from all 88 selected donkeys. Three genera of ticks were identified, namely Rhipicephalus (97.2%; 486/500), Amblyomma (2.2%; 11/500), and Hyalomma (0.6%; 3/500). Seven species were identified, of which Rhipicephalus evertsi evertsi with 50.4% (252/500) was the most prevalent, followed by Rhipicephalus appendiculatus (27.4%; 137/500), Rhipicephalus turanicus (11.6; 10/500), Rhipicephalus (boophilus) microplus (6.8; 20/500), Amblyomma hebraeum (2.2%; 11/500), Rhipicephalus sanguineus (1%; 5/500) and Hyalomma truncatum (0.6%; 3/500). Rhipicephalus evertsi evertsi occurred in all locations, whereas Hyalomma truncatum occurred only in the Boane district. Males were the most prevalent (67.2%; 336/500). The study revealed that donkeys in Maputo Province were infested with seven tick species of which R. evertsi evertsi was the main species.

     

Comparison of resistance in three breeds of cattle against African ixodid ticks

Tick resistance in three breeds of cattle, two indigenous breeds (Arssi and Boran) and one Boran x Friesian cross-breed, were compared following natural tick infestations at Abernossa ranch in Ethiopia. The local Arssi breed was found to have the highest tick resistance, followed by the Boran breed, whereas the Boran x Friesian was the least resistant. Over a period of 12 months, from October 1991 to September 1992, a total of 32 897 ticks composed of four genera were collected from the animals. The four most abundant tick species were Amblyomma variegatum (61.7%), Boophilus decoloratus (16%); Rhipicephalus evertsi evertsi (16.3%) and Hyalomma marginatum rufipes (3.7%). Furthermore, 63.5% of all ticks were collected from cross-breed cattle, and 26.2% from the Boran, whereas the local Arssi breed carried only 10.3%. The results indicated that cattle resistant to one species of tick were also resistant to other tick species.     

Studies to evaluate the effectiveness of sex pheromone-impregnated formulations for control of populations of the American dog tick,Dermacentor variabilis (Say) (Acari: Ixodidae)

The sex pheromone, 2,6-dichlorophenol, was combined with a pesticide to control populations of the American dog tick,Dermacentor variabilis (Say). This pheromone persisted in the fur of treated dogs for at least 18 days. The mixture of pheromone and pesticide was much more effective in reducing mating among the surviving ticks than the treatments without pheromone. The pheromone-pesticide mixture also killed significantly more ticks than the treatment without pheromone. This increased effectiveness was due almost entirely to the significantly greater kill of male ticks. Combining the sex pheromone with pesticide treatments offers a means of suppressing tick populations by curtailing their mating and subsequent reproductive success. The potential applications of this technique and the benefits to be expected are discussed.     

Pheromone/acaricide mixtures in the control of the tickAmblyomma hebraeum: Effects of acaricides on attraction and attachment

Unfed adults and nymphs of the bont tickAmblyomma hebraeum Koch are attracted to hosts on which fed males, emitting an aggregation-attachment pheromone (AAP), are attached. Pheromone/acaricide mixtures have the potential to selectively attract and kill these ticks. We have investigated the effects of three acaricides, amitraz (an amidine), flumethrin (a synthetic pyrethroid) and chlorfenvinphos (an organophosphate), combined with AAP, on the attraction and attachment of the unfed adults. Attraction, measured in field assays involving AAP and CO₂, was not inhibited by any of the acaricides. Attachment was measured over 24 h on the ears of rabbits which had been treated with AAP and mixtures of AAP and the acaricides. None of the acaricides inhibited initial attachment. Flumethrin caused rapid and high mortality in attached and unattached ticks. Chlorfenvinphos caused little mortality in the first 24 h on the ears of the rabbits, but 75% of the exposed ticks died over the next 14 days while held in an incubator. Amitraz caused the ticks to detach after an initial period of attachement; there was little mortality in the detached ticks over the next two weeks and they could later be induced to re-attach to other rabbits. Flumethrin was considered to be the compound of choice for use in pheromone/acaricide mixtures.     

Ticks and Tick-Borne Diseases of Livestock Belonging to Resource-poor Farmers in the Eastern Free State of South Africa

The paper provides a summary of three studies conducted in the eastern Free State of South Africa between 1998 and 2000. In a questionnaire-based study approximately 21% of interviewed resource-poor farmers (n = 150) indicated that they experienced problems with ticks and tick-borne diseases. About 56% of farmers indicated that tick-related problems were most severe in summer, while 32% indicated that the most problems were encountered in winter. About 12% indicated that the tick problems were experienced throughout the year. Farmers also indicated that the highest tick burdens were experienced between spring and early winter. The principal ticks infesting cattle (n = 30) were found to be Boophilus decoloratus (53.1%), Rhipicephalus evertsi evertsi (44.7%), Rhipicephalus follis (1.0%), Rhipicephalus gertrudae (0.7%) and Rhipicephalus warburtoni (0.4%). On small stock (n = 188), R. evertsi evertsi (68%) and B. decoloratus (32%) were recorded as the main ticks in the study area. A sero-epidemiological survey of cattle (n = 386) showed that 94% of cattle were sero-positive for Babesia bigemina by IFAT, while 87% were sero-positive for Anaplasma by indirect ELISA. All the animals were sero-negative for Babesia bovis and this is probably because the tick vector, Boophilus microplus, is not present in the study area. All sheep and goats were sero-positive for Theileria species by IFAT while 85% of sheep and 100% of goats tested positive for Anaplasma species by competition inhibition ELISA. The high incidence of positive serological results for B. bigemina and Anaplasma in cattle, and Theileria and Anaplasma in sheep and goats and the absence of clinical cases would indicate that this area is endemically stable for these diseases. This revised version was published online in July 2006 with corrections to the Cover Date.