Ixodid ticks feeding on humans in South Africa: with notes on preferred hosts,geographic distribution,seasonal occurrence and transmission of pathogens

This paper records the identities of 558 ixodid ticks feeding on 194 humans in South Africa. These ticks belonged to 20 species in six genera and those most frequently encountered were Amblyomma hebraeum, Haemaphysalis leachi, Hyalomma marginatum rufipes, Hyalomma truncatum, Rhipicephalus appendiculatus, Rhipicephalus gertrudae and Rhipicephalus simus. With the exception of the larvae of R. appendiculatus, the incidents of these ticks feeding on humans correlated well with their seasonal occurrences on preferred hosts. Ticks were also collected at monthly intervals, for 14 consecutive months, from the clothing of a game-guard providing protection for field-workers engaged in the collection, by means of flannel strips, of free-living ticks from the vegetation of four localities in the southern region of the Kruger National Park. In addition, with the exception of 3 months when the particular worker was absent, ticks that fed on one of the field-workers were collected over the same 14-month period. A total of 54,429 free-living ticks belonging to 14 species and six genera were collected from the vegetation at the four localities during this time and 3751 ticks belonging to 11 species and six genera from the clothing of the game-guard. The larvae of A. hebraeum and Boophilus decoloratus were the most numerous of the immature ticks, and H. leachi and R. simus of the adults on both the vegetation and the guard’s clothing. Ticks fed on the field-worker on six occasions and 14 were collected, all of which were A. hebraeum larvae. This revised version was published online in July 2006 with corrections to the Cover Date.     

Collection of Free-Living Nymphs and Adults of Amblyomma Hebraeum (Acari: Ixodidae) with Pheromone/Carbon Dioxide Traps at 5 Different Ecological Sites in Heartwater Endemic Regions of South Africa

The capture of free-living adults and nymphs of Amblyomma hebraeum, the main vector of heartwater in domestic and wild ruminants in South Africa, by means of attraction–aggregation–attachment–pheromone/carbon dioxide traps at five endemic localities in South Africa is described. Although the traps were used successfully at each of the localities, no determination of their efficiency in relation to the actual abundance of ticks at a particular site was made. This study confirmed that the traps could be used in a variety of ecological areas to locate populations of free-living adult A. hebraeum.     

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.     

Field sampling of unfed adults ofAmblyomma hebraeum Koch

No methods have previously been available for the field sampling of unfed adults ofAmblyomma hebraeum. We released 2000 unfed adults into a 5-m² area in mopane woodland in southeastern Zimbabwe. The ticks sought shelter beneath the debris on the soil surface but emerged and became active in host seeking when stimulated. Effective stimuli were the odours of cattle and sheep and high concentrations of carbon dioxide from dry ice. The ticks did not respond to low concentrations of carbon dioxide, vibrations generated by stamping the ground with poles, or two volatile components of ox breath known to attract tsetse flies. The presence of humans in the release area also had little or no effect in stimulating the ticks. As adults ofA. hebraeum are large and conspicuous they can easily be collected from the soil surface, either manually or using a vacuum device, after stimulation by cattle or the release of high concentrations of carbon dioxide.     

Detection of lumpy skin disease virus in saliva of ticks fed on lumpy skin disease virus-infected cattle

Lumpy skin disease is an economically important disease of cattle that is caused by the lumpy skin disease virus (LSDV), which belongs to the genus Capripoxvirus. It is endemic in Africa and outbreaks have also been reported in the Middle-East. Transmission has mostly been associated with blood-feeding insects but recently, the authors have demonstrated mechanical transmission by Rhipicephalus appendiculatus as well as mechanical/intrastadial and transstadial transmission by Amblyomma hebraeum. Saliva is the medium of transmission of pathogens transmitted by biting arthropods and, simultaneously, it potentiates infection in the vertebrate host. This study aimed to detect LSDV in saliva of A. hebraeum and R. appendiculatus adult ticks fed, as nymphs or as adults, on LSDV-infected animals, thereby also demonstrating transstadial or mechanical/intrastadial passage of the virus in these ticks. Saliva samples were tested for LSDV by real-time PCR and virus isolation. Supernatants obtained from virus isolation were further tested by real-time PCR to confirm that the cytopathic effects observed were due to LSDV. Lumpy skin disease virus was detected, for the first time, in saliva samples of both A. hebraeum and R. appendiculatus ticks. At the same time, mechanical/intrastadial and transstadial passage of the virus was demonstrated and confirmed in R. appendiculatus and A. hebraeum.     

Detection of “<Emphasis Type="Italic">Candidatus</Emphasis> Rickettsia sp. strain Argentina”and <Emphasis Type="Italic">Rickettsia bellii</Emphasis> in <Emphasis Type="Italic">Amblyomma</Emphasis> ticks (Acari: Ixodidae) from Northern Argentina

Ixodid ticks were collected from vegetation and from humans, wild and domestic mammals in a rural area in the semi-arid Argentine Chaco in late spring 2006 to evaluate their potential role as vectors of Spotted Fever Group (SFG) rickettsiae. A total of 233 adult ticks, identified as Amblyomma parvum, Amblyomma tigrinum and Amblyomma pseudoconcolor, was examined for Rickettsia spp. We identified an SFG rickettsia of unknown pathogenicity, “Candidatus Rickettsia sp. strain Argentina”, in A. parvum and A. pseudoconcolor by PCR assays targeting gltA, ompA, ompB and 17-kDa outer membrane antigen rickettsial genes. Rickettsia bellii was detected in a host-seeking male of A. tigrinum. Amblyomma parvum is widespread in the study area and is a potential threat to human health.     

The effect of rainfall on tick challenge at Kyle Recreational Park,Zimbabwe

The effect of rainfall pattern on tick challenge was investigated at Kyle Recreational Park, Zimbabwe, from 1991 to 1992 using drag and removal plot methods to sample environmental tick density. The abundance of adults and nymphs of the brown ear-tick Rhipicephalus appendiculatus and larvae of the bont tick Amblyomma hebraeum was positively correlated with monthly rainfall, whereas no relationship with rainfall was revealed for larval R. appendiculatus, adults of the red-legged tick R. evertsi, or larvae of the blue tick Boophilus decoloratus. A comparison between 1991 (490 mm rainfall) and the drought year of 1992 (161 mm) revealed significant differences in the abundance of R. appendiculatus, A. hebraeum, and B. decoloratus. During the wet season, R. appendiculatus adults were 2–3 time more numerous in the environment during the higher rainfall year of 1991. A. hebraeum larval abundance exhibited a similar pattern to that of R. appendiculatus adults, but B. decoloratus larvae were more abundant in the drought year of 1992 during both the wet and dry seasons. Comparable tick abundance data collected at Kyle during the above-average rainfall years of 1975–1977 (mean = 1029 mm) were compared with tick challenge during the below-average rainfall years of 1991–1992 (mean = 326 mm). In grassland sand habitat and all habitats combined R. appendiculatus adults, nymphs, and larvae were much more abundant during the high rainfall years. In contrast, larvae of B. decoloratus were more numerous during the drier years. A. hebraeum larvae were also more abundant during the drier years. The strong positive correlation of adult R. appendiculatus abundance with rainfall and the coincidence of increased adult tick challenge with increased rainfall indicates that adult R. appendiculatus tick burden on hosts would be heaviest during the wet season and high rainfall years.     

Tick prevalence and species diversity on Aldabran giant tortoises (Dipsochelys dussumieri) in relation to host range and host size in a restored ecosystem, Kenya

Tick species density and diversity on Aldabran tortoises was investigated in relation to the habitat range and size of each tortoise. Identification of tick infestation patterns forms an important aspect of effective tick control. Ten Aldabran tortoises were de‐ticked and monitored over the course of 2 months. Tick species found were Amblyomma sparsum, Amblyomma nuttalli, Amblyomma hebraeum and Boophilus decoloratus, with the most prevalent species found being A. sparsum. Tick loads varied considerably from 20 to 214 ticks per tortoise, with most ticks collected from the head/neck region. Tortoises ranging outside Haller Park had higher tick loads (70–214) compared with tortoises ranging within Haller Park (20–99). Tick load was not correlated with tortoise size. Results indicate that tick loads are related to the habitat range of the tortoises and may indirectly also be related to food preference and host food availability. Implications of the findings and appropriate tick control measures are discussed.     

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.     

Candidatus Rickettsia andeanae,a spotted fever group agent infecting Amblyomma parvum ticks in two Brazilian biomes

Adult ticks of the species Amblyomma parvum were collected from the vegetation in the Pantanal biome (state of Mato Grosso do Sul) and from horses in the Cerrado biome (state of Piauí) in Brazil. The ticks were individually tested for rickettsial infection via polymerase chain reaction (PCR) targeting three rickettsial genes, gltA, ompA and ompB. Overall, 63.5% (40/63) and 66.7% (2/3) of A. parvum ticks from Pantanal and Cerrado, respectively, contained rickettsial DNA, which were all confirmed by DNA sequencing to be 100% identical to the corresponding fragments of the gltA, ompA and ompB genes of Candidatus Rickettsia andeanae. This report is the first to describe Ca. R. andeanae in Brazil.     

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.     

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.     

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.

     

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. 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.     

The impact of RNA interference of the subolesin and voraxin genes in male <Emphasis Type="Italic">Amblyomma hebraeum</Emphasis> (Acari: Ixodidae) on female engorgement and oviposition

Reducing or replacing the use of chemical pesticides for tick control is a desirable goal. The most promising approach would be to develop vaccines that protect hosts against tick infestation. Antigens suitable for the development of anti-tick vaccines will likely be those essential for vital physiological processes, and in particular those directly involved in feeding and reproduction. In this study genes from Amblyomma hebraeum Koch that encode for subolesin and voraxin were studied in male ticks by RNA interference (RNAi). Males (unfed or fed) were injected with dsRNA of (1) subolesin, (2) voraxin, (3) subolesin plus voraxin or (4) injection buffer, after which they were held off-host overnight and then allowed to feed on rabbits together with normal female A. hebraeum. Females that fed together with male ticks injected with subolesin or subolesin + voraxin dsRNA had a higher rate of mortality, weighed substantially less and produced a smaller egg mass than the controls. However, females feeding with males injected with voraxin dsRNA alone were not significantly different from the controls with respect to mortality, engorged weight or fecundity. However, as assessed by semi-quantitative RT-PCR, voraxin was not silenced in this study, the reasons for which remain unknown. The results of this study suggest that A. hebraeum subolesin is worthy of further testing as a candidate tick vaccine antigen.     

Rickettsia amblyommii infecting Amblyomma sculptum in endemic spotted fever area from southeastern Brazil

The Rickettsia bacteria include the aetiological agents for the human spotted fever (SF) disease. In the present study, a SF groupRickettsia amblyommii related bacterium was detected in a field collected Amblyomma sculptum (Amblyomma cajennense species complex) tick from a Brazilian SF endemic site in southeastern Brazil, in the municipality of Juiz de Fora, state of Minas Gerais. Genetic analysis based on genes ompA,ompB and htrA showed that the detected strain, named R. amblyommii str. JF, is related to the speciesR. amblyommii.     

Molecular detection of Rickettsia aeschlimannii in Hyalomma spp. ticks from camels (Camelus dromedarius) in Nigeria,West Africa

Several species of the spotted fever group rickettsiae have been identified as emerging pathogens throughout the world, including in Africa. In this study, 197 Hyalomma ticks (Ixodida: Ixodidae) collected from 51 camels (Camelus dromedarius) in Kano, northern Nigeria, were screened by amplification and sequencing of the citrate synthase (gltA), outer membrane protein A (ompA) and 17‐kDa antigen gene fragments. Rickettsia sp. gltA fragments were detected in 43.3% (42/97) of the tick pools tested. Rickettsial ompA gene fragments (189 bp and 630 bp) were detected in 64.3% (n = 27) and 23.8% (n = 10) of the gltA‐positive tick pools by real‐time and conventional polymerase chain reaction (PCR), respectively. The amplicons were 99–100% identical to Rickettsia aeschlimannii TR/Orkun‐H and R. aeschlimannii strain EgyRickHimp‐El‐Arish in GenBank. Furthermore, 17‐kDa antigen gene fragments of 214 bp and 265 bp were detected in 59.5% (n = 25) and 38.1% (n = 16), respectively, of tick pools, and sequences were identical to one another and 99–100% identical to those of the R. aeschlimannii strain Ibadan A1 in GenBank. None of the Hyalomma impressum ticks collected were positive for Rickettsia sp. DNA. Rickettsia sp. gltA fragments (133 bp) were detected in 18.8% of camel blood samples, but all samples were negative for the other genes targeted. This is the first report to describe the molecular detection of R. aeschlimannii in Hyalomma spp. ticks from camels in Nigeria.     

Salivary gland degeneration and vitellogenesis in the ixodid tick Amblyomma hebraeum: Surpassing a critical weight is the prerequisite and detachment from the host is the trigger

The normal engorged body weight of female ixodid ticks (Acari: Ixodidae) is about 100× the unfed weight. Virgin female Amblyomma hebraeum normally do not feed beyond 10× the unfed weight. However, about 10–20% of a population of virgins will feed to perhaps 20× the unfed weight, but not much beyond that. In A. hebraeum, when females surpass about 10× the unfed weight, the following changes in physiology occur if they are removed from the host: (a) they will not reattach if given the opportunity, (b) their salivary glands (SGs) will undergo autolysis within 4 days if they are mated or 8 days if they are virgin, and (c) egg maturation and oviposition will occur in due course. Mated or virgin female ticks removed from the host below about 10× the unfed weight do not experience the latter changes (Kaufman, W.R., Lomas, L., 1996. ‘Male Factors’ in ticks: their role in feeding and egg development. Invertebrate Reproduction and Development 30, 191–198). In 1984 we named this transitional weight, the ‘critical weight’ (CW). Its absolute value is probably a species-specific characteristic (Kaufman, W.R., 2007. Gluttony and sex in female ixodid ticks: how do they compare to other blood-sucking arthropods? Journal of Insect Physiology 53, 264–273). Although mated females tend to engorge within a day of surpassing the CW, virgin females surpassing the CW can remain attached to the host for at least several weeks more. It is not known whether the physiological changes in the SGs and ovaries listed above occur in those large virgins that remain attached, although we suppose that this would be maladaptive. Instead, we hypothesize in this study that surpassing the CW is only a prerequisite for inducing these changes, and that detachment is the actual trigger. We support our hypothesis by demonstrating that large virgins, remaining attached to a host for 8 days, did not undergo SG degeneration nor complete egg maturation during the attachment period. Those changes occurred only within 8 days following detachment. So some type of sensory information associated with attachment to the host, and still undefined, inhibits expression of the physiological changes hitherto associated merely with surpassing the CW.     

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.