Ticks (Acari: Ixodidae) in the Serra da Canastra National Park in Minas Gerais,Brazil: species,abundance, ecological and seasonal aspects with notes on rickettsial infection

The Cerrado Biome is the second largest in Brazil covering roughly 2 million km², with varying features throughout its area. The Biome is endangered but it is also source of animal species for rural, green urban and degraded rainforest areas. Ticks are among Cerrado species that establish at anthropogenic sites and although information about them is steadily increasing, several features are unknown. We herein report tick species, abundance and some ecological relationships within natural areas of the Cerrado at higher altitudes (800–1500 m) within and around Serra da Canastra National Park, in Minas Gerais State Brazil. In total of 1196 ticks were collected in the environment along 10 campaigns held in 3 years (2007–2009). Amblyomma sculptum was the most numerous species followed by Amblyomma dubitatum and Amblyomma brasiliense. Distribution of these species was very uneven and an established population of A. brasiliense in the Cerrado is reported for the first time. Other tick species (Amblyomma ovale, Amblyomma nodosum, Amblyomma parvum, Ixodes schulzei and Haemaphysalis leporispalustris) were found in lesser numbers. Domestic animals displayed tick infestations of both rural and urban origin as well as from natural areas (Rhipicephalus sanguineus sensu lato, Rhipicephalus microplus, Dermacentor nitens, A. sculptum, A. ovale, Amblyomma tigrinum, Argas miniatus). Amblyomma sculptum had the widest domestic host spectrum among all tick species. DNA of only one Rickettsia species, R. bellii, was found in an A. dubitatum tick. Several biological and ecological features of ticks of the studied areas are discussed.     

Ixodid ticks of African buffalo (<Emphasis Type="Italic">Syncerus caffer</Emphasis>), impala (<Emphasis Type="Italic">Aepyceros melampus</Emphasis>) and elephant (<Emphasis Type="Italic">Loxodonta africana</Emphasis>) in five protected park estates in the Zambezi valley,Zimbabwe

Wildlife hosts many pathogens of economic importance and is considered as a reservoir of important tick-borne diseases of livestock in southern Africa. The species composition of ticks parasitizing buffalo (Syncerus caffer), impala (Aepyceros melampus) and elephant (Loxodonta africana) was investigated in five protected parks in the Zambezi valley, Zimbabwe. A total of 1104 adult ticks was collected from 75 adult animals comprising five buffaloes, five elephants and five impalas drawn from five protected wildlife parks. Five tick species belonging to two genera were recovered, with Rhipicephalus decoloratus being the most prevalent species in all the three animal groups. Amblyomma hebraeum was only recovered from buffaloes whereas Rhipicephalus zambeziensis was recovered from buffalos and elephants. Significant differences in mean tick species distribution and concentration were observed amongst the wildlife parks and these appeared to be influenced by the number of hosts in each park. The study revealed that buffaloes are the major host of R. decoloratus in the Zambezi valley. The presence of these ixodid ticks within the Zambezi valley may have significant ecological and economic impacts on wildlife conservation, domestic animals and human health.     

Phylogenetic insights on Mediterranean and Afrotropical <Emphasis Type="Italic">Rhipicephalus</Emphasis> species (Acari: Ixodida) based on mitochondrial DNA

This study was performed to determine the tick species that infest cattle and humans throughout an altitudinal gradient in the Yungas Biogeographic Province of Argentina. The presence of tick-borne bacteria of the genera Rickettsia, Ehrlichia and Borrelia in the collected ticks was also evaluated. Samples of ticks parasitizing cattle and humans were carried out in different seasons. Questing ticks (adults and nymphs) were collected from vegetation and analyzed to detect the presence of Rickettsia, Ehrlichia and Borrelia by a battery of different PCRs. Five species of hard ticks were found parasitizing cattle: Amblyomma sculptum, Amblyomma tonelliae, Amblyomma hadanii, Haemaphysalis juxtakochi and Ixodes pararicinus. Amblyomma sculptum (immature and adults), A. tonelliae (immature and adults), A. hadanii (larvae) and one nymph of I. pararicinus were found attached to humans. Rickettsia amblyommatis was detected in one nymph of A. hadanii. DNA of a Borrelia genospecies belonging to the B. burgdorferi s.l. complex (phylogenetically related to haplotypes previously reported in Ixodes aragaoi from Uruguay and I. pararicinus from Argentina) was detected in adults of I. pararicinus. Amblyomma sculptum and I. pararicinus appear to be the tick species more frequent on cattle in the YBP from Argentina, and A. sculptum and A. tonelliae, were the main ticks found attached to humans. The medical importance of the bacteria of the genus Rickettsia and Borrelia detected in this work remains unknown.     

Distribution of Spotted Fever Group Rickettsiae in Hard Ticks (Ixodida: Ixodidae) from Panamanian Urban and Rural Environments (2007–2013)

Tick-borne rickettsiosis is an important emerging disease in Panama; to date, there have been 12 confirmed cases, including eight fatalities. To evaluate the distribution of rickettsiae in Panamanian ticks, we collected questing and on-host ticks in urban and rural towns in elevations varying between 0 and 2300 m. A total of 63 sites (13 urban and 50 rural towns) were used to develop models of spatial distributions. We found the following tick species: Rhipicephalus sanguineus s.l. (present in 54 of 63 towns and cities), Amblyomma mixtum (45/63), Dermacentor nitens (40/63), A. ovale (37/63), Rhipicephalus microplus (33/63), A. oblongoguttatum (33/63), Ixodes affinis (3/63), and Ixodes boliviensis (2/63). Rhipicephalus sanguineus s.l. was present in urban and rural towns, and other species were present only in rural towns. DNA was extracted from 408 R. sanguineus s.l., 387 A. mixtum, 103 A. ovale, and 11 A. oblongoguttatum and later tested for rickettsiae genes using PCR. Rickettsia DNA was detected in ticks from 21 of 63 localities. Rickettsia rickettsii was detected in five A. mixtum (1.29%), and Candidatus “Rickettsia amblyommii” was found in 138 A. mixtum (35%), 14 R. sanguineus (3.4%), and one A. ovale (0.9%). These results suggest that much of rural Panama is suitable for the expansion of tick populations and could favor the appearance of new tick-borne rickettsiosis outbreaks.     

Prevalence and spatial distribution of Ixodid tick populations in the forest fringes of Western Ghats reported with human cases of Kyasanur forest disease and monkey deaths in South India

Kyasanur forest disease (KFD) is a major tick-borne viral haemorrhagic fever caused by KFD virus (KFDV) (Flaviviridae). The disease was reported to be confined to five districts of Karnataka state India until 2011. During 2012–2016, emergence of KFD has been reported in newer areas of Karnataka and adjoining states. Therefore, survey of tick vectors was carried out in these new areas of Karnataka and adjoining states reported with monkey deaths and human cases of KFD. In all selected sites, ticks from the forest floor were collected by lint clothes using flagging method. Tick samples were tested for KFDV nucleic acid by real-time RT-PCR. A total of 4772 ticks, comprising eight species of genus Haemaphysalis and one species each of genus Amblyomma, Ixodes and Rhipicephalus was collected. Haemaphysalis spinigera, the principal vector of KFDV was the predominant tick species (59.5%) collected followed by H. turturis (8.6%). The abundance of H. spinigera ranged from 9.2 to 33.9 per man-hour in the six districts surveyed. Of 214 (4418 tick samples) pools screened by real-time RT-PCR, two pools of H. spinigera were positive for KFDV. High abundance of Haemaphysalis vectors in the six districts indicated that the districts are receptive for KFD outbreaks. KFDV was detected in the tick vectors in the new foci of the KFD. Data on tick distribution will be useful in creating KFD risk map for strengthening the ongoing preventive measures such as vaccination and supply of insect repellents to the high risk groups and intensive health education.     

Ixodid ticks (Parasitiformes: Ixodidae) from small mammals in reforesting boreal habitats of the northern European Russia

Abundance of 2 ixodid tick species, Ixodes (Exopalpiger) trianguliceps Birula, 1895 and Ixodes (Ixodes) persulcatus Schulze, 1930 was studied during 6 years, in 1998–2001 and 2003–2004, at a research station in Gomselga Village (central Karelia, Kondopoga District, 62°04′N, 33°55′E). New data on host species composition and ixodid tick abundance were obtained from 4 forest plots at different stages of reforestation (secondary succession) following felling that occurred 7–14, 12–19, 25–32, and 80–87 years ago. Individuals of I. persulcatus predominated and constituted 73% of the total ticks collected. The occurrence rates of different developmental stages were shown to fluctuate in the course of reforestation both in I. trianguliceps (larvae, 2.8–5.3; nymphs, 1.5–2.2; adults, 0–0.09) and in I. persulcatus (larvae, 4.3–10.6; nymphs, 0.6–4.2).     

In vitro efficacy of <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> sensu lato against unfed <Emphasis Type="Italic">Amblyomma parvum</Emphasis> (Acari: Ixodidae)

Amblyomma parvum Aragão (Acari: Ixodidae) is a tick species found with wide distribution in the Neotropical region. Even though it is a wildlife-related tick, it is also a frequent parasite of domestic animals, is aggressive to human beings and may harbor pathogenic microorganisms. Therefore, it is a target species for control on domestic animals, particularly those at the rural–wildlife interface. Herein, the efficacy of two isolates (E9 and IBCB 425) of an entomopathogenic fungus, Metarhizium anisopliae sensu lato, already evaluated for ticks that parasitize domestic animals, was tested against unfed A. parvum adults. Both isolates displayed high acaricidal efficacy after immersion in fungal conidial suspensions for 5 min. Isolate E9 killed all ticks by the 7th day post-treatment, and isolate IBCB 425 did so by the 11th day. Tick mortality of 80 and 90% was achieved as early as the 3rd and 4th days, respectively, with both treatments. Thus, if a commercial M. anisopliae s.l. acaricide against domestic animal ticks is developed, it would also be effective against A. parvum.     

Detection of <Emphasis Type="Italic">Rickettsia</Emphasis> spp. in ticks associated to wild mammals in Northeastern Brazil,with notes on an undetermined <Emphasis Type="Italic">Ornithodoros</Emphasis> sp. collected from marsupials

We report tick infestations and rickettsial detection in ticks infesting free-living wild mammals (Monodelphis domestica, Tolypeutes tricinctus, Thrichomys inermis and Kerodon rupestris) captured in the Caatinga ecoregion of Bahia state, northeastern Brazil, during September to December 2016. Overall, 117 ticks (61 larvae, 25 nymphs, 25 males, 6 females) belonging to two genera, and at least three species were collected: Amblyomma auricularium, Amblyomma parvum, Amblyomma sp., Ornithodoros rietcorreai and an unidentified Ornithodoros sp. We provide new host records to the rodent T. inermis parasitized by larva and nymphs of A. auricularium and to the marsupial M. domestica infested by larvae of A. auricularium. Furthermore, we describe new tick-host association for larvae of O. rietcorreai on the rodents K. rupestris and T. inermis. Concerning tick-Rickettsia associations, we detected Rickettsia amblyommatis and an uncharacterized species of Rickettsia belonging to the spotted fever group (SFG) in both A. auricularium and A. parvum. Additionally, ‘Candidatus Rickettsia andeanae’ was detected in A. parvum as well.     

Detection of <Emphasis Type="Italic">Rickettsia</Emphasis> spp. in ticks parasitizing toads (<Emphasis Type="Italic">Rhinella marina</Emphasis>) in the northern Brazilian Amazon

This study evaluated rickettsial infection in ticks collected on toads from the northern Brazilian Amazon (Amapá state), where to our knowledge there are neither records of ticks from amphibians nor rickettsial infections in ticks. During 2016–2017, a total of 22 free-living toads were captured and identified as Rhinella marina. Overall, 12 (54.5%) toads were parasitized by a total of 97 ticks (6 males, 39 females, 31 nymphs, 21 larvae), giving a mean intensity of 8.1 ticks per infested toad. Two tick species were morphologically identified: Amblyomma rotundatum Koch (31 females, 14 nymphs), and Amblyomma dissimile Koch (6 males, 8 females, 17 nymphs). The 21 larvae were morphologically denoted as Amblyomma sp. Five toads were co-infested by A. rotundatum and A. dissimile. Morphological identifications were confirmed by nucleotide sequencing of fragments of the mitochondrial genes 16S rDNA, 12S rDNA and/or COX1. A total of 54 ticks were analyzed for the presence of rickettsial DNA. Eleven (9 females and 2 nymphs) out of 14 A. rotundatum ticks contained Rickettsia bellii. None of the 25 specimens of A. dissimile (6 males, 6 females, 13 nymphs) contained amplifiable rickettsial DNA. From 15 Amblyomma sp. larvae, a pool of 10 individuals contained Rickettsia sp. strain Colombianensi. Sequencing of the 16S rDNA amplicon derived from the positive pool yielded a sequence of A. dissimile. We detected Rickettsia sp. strain Colombianensi for the first time in Brazil. Prior records of this agent were restricted to Colombia and Honduras. In addition, we report the presence of A. rotundatum for the first time in the state of Amapá, where the only other record of A. dissimile was registered over 20 years ago.     

PCR identification and phylogenetic analysis of the medically important dust mite <Emphasis Type="Italic">Suidasia medanensis</Emphasis> (Acari: Suidasiidae) in Malaysia

Hard ticks are a perennial problem in livestock systems throughout Mexico. No data are currently available on tick prevalence, infestation intensity and population dynamics for sheep in the humid tropics of Mexico. Blackbelly sheep (n = 30) in Tabasco state, Mexico, were examined every 3 weeks for 1 year, and all Ixodidae ticks were counted and removed for analysis. Tick species were identified, infestation prevalence and intensity calculated per animal, and infestation per body zone determined. Overall infestation on the studied animals was 51.9%. Four tick species were identified (Amblyomma mixtum, A. imitator, Rhipicephalus microplus and R. annulatus), the most abundant being A. mixtum (94.9%). Prevalence was highest during the northwinds season (61.9%), followed by the dry (48.5%) and rainy (47.3%) seasons; however, the intensity (2.01–2.07 ticks/sheep) did not differ between seasons. Infestation was most frequent in certain zones including the axilla, crotch, udder, abdomen, thorax and rib area. Sheep in the humid tropics of Tabasco, Mexico, are parasitized by four hard tick species and prevalence is highest in the northwinds season. The present data constitute an important baseline for developing sustainable tick control programs for sheep in the humid tropics of Mexico.     

Birds as hosts of ixodid ticks (Acarina,Ixodidae)

rophic relations between ixodid ticks of the world fauna parasitizing as adults on birds are analyzed. These ticks include 60 out of 650 species of the family Ixodidae and belong to the genera Ixodes, Haemaphysalis, and Amblyomma. Relations between birds and ticks of the genus Ixodes are most ancient. Colonial marine birds seem to be the first hosts of ixodid ticks in the class of birds. Trophic specialization of ticks is not associated with bird taxa, because in most cases no tick specificity is caused by either phylogenetic features of hosts, or specific environmental conditions.     

Ticks collected on birds in the state of São Paulo,Brazil

The present study reports a collection of Amblyomma spp. ticks in birds from several areas of the state of São Paulo, Brazil. A total of 568 tick specimens (404 larvae, 164 nymphs) were collected from 261 bird specimens. From these ticks, 204 (36%) specimens (94 larvae, 110 nymphs) were reared to the adult stage, being identified as Amblyomma longirostre (94 larvae, 90 nymphs), Amblyomma calcaratum (13 nymphs), Amblyomma nodosum (2 nymphs), and Amblyomma cajennense (5 nymphs). Additionally, 39 larvae reared to the nymphal stage and 8 nymphs that died before reaching the adult stage were identified as A. longirostre according to peculiar characters inherent to the nymphal stage of this species: scutum elongate, and hypostome pointed. The remaining 271 larvae and 46 nymphs were identified as Amblyomma sp. Ticks were collected from 51 species of birds distributed in 22 bird families and 6 orders. The order Passeriformes constituted the vast majority of the records, comprising 253 (97%) out of the 261 infested birds. Subadults of A. longirostre were identified from 35 species of Passeriformes, comprising 11 families (Cardinalidae, Dendrocolaptidae, Fringillidae, Furnariidae, Parulidae, Pipridae, Thamnophilidae, Thraupidae, Turdidae, Tyrannidae, and Vireonidae), and from 1 species of a non-passerine bird, a puffbird (Bucconidae). Subadults of A. calcaratum were identified from 5 species of Passeriformes, comprising 5 families (Cardinalinae, Conopophagidae, Pipridae, Thamnophilidae and Turdidae). Subadults of A. nodosum were identified from 2 species of Passeriformes, comprising two bird families (Thamnophilidae and Pipridae). Subadults of A. cajennense were identified from 2 species of non-passerine birds, belonging to 2 different orders (Ciconiiformes: Threskiornithidae, and Gruiformes: Cariamidae). Birds were usually infested by few ticks (mean infestation of 2.2 ticks per bird; range: 1–16). Currently, 82 bird species are known to be infested by immature stages of A. longirostre, with the vast majority [74 (90%)] being Passeriformes. Our results showed that Passeriformes seems to be primary hosts for subadult stages of A. longirostre, A. calcaratum, and A. nodosum. However, arboreal passerine birds seem to be the most important hosts for A. longirostre whereas ground-feeding passerine birds seem to be the most important for both A. calcaratum and A. nodosum. In contrast, the parasitism of birds by subadults of A. cajennense has been restricted to non-passerine birds.     

Ticks on captive and free-living wild animals in northeastern Brazil

From 2005 to 2009, 147 ticks found on 32 wild animals from or referred to two zoobotanical parks (Parque Zoobotânico Arruda Câmara and Parque Estadual Dois Irmãos) located in northeastern Brazil were identified. Ticks found on two veterinarians working in one of the parks (i.e., Parque Estadual Dois Irmãos), after return from forested areas within the park’s territory, were also identified. The following tick-host associations were recorded: Amblyomma fuscum Neumann on Boa constrictor L.; Amblyomma longirostre Koch on Ramphastos vitellinus ariel Vigors and Coendou prehensilis (L.); Amblyomma varium Koch on Bradypus variegates Schinz; Amblyomma rotundatum Koch on Chelonoidis carbonaria (Spix), Chelonoidis denticulata (L.), Micrurus ibiboboca (Merrem), Python molurus bivittatus Kuhl, Iguana iguana (L.) and B. variegatus; Amblyomma nodosum Neumann on Myrmecophaga tridactyla L. and Tamandua tetradactyla (L.); and Rhipicephalus sanguineus (Latreille) on Nasua nasua (L.). The ticks found on the veterinarians were identified as nine Amblyomma larvae. The presence of Am. nodosum in Pernambuco and Am. rotundatum and Am. varium in Paraíba is recorded for the first time and the occurrence of Am. longirostre in Pernambuco is confirmed. Ramphastos vitellinus ariel is a new host record for Am. longirostre whereas M. ibiboboca and B. variegatus are new host records for Am. rotundatum. Finally, the human parasitism by Amblyomma ticks is reported for the first time in Pernambuco, highlighting the potential of tick-borne pathogen transmission in this state.     

First molecular detection of <Emphasis Type="Italic">Anaplasma phagocytophilum</Emphasis> in the hard tick <Emphasis Type="Italic">Rhipicephalus haemaphysaloides</Emphasis> in Taiwan

Anaplasma phagocytophilum is transmitted mainly by hard ticks and can cause potentially fatal granulocytic anaplasmosis in humans, but its occurrence in ticks in Taiwan has never been investigated although this pathogen has been detected in Taiwanese rodents before. Ticks collected from small mammals in Hualien, eastern Taiwan, were assayed for Anaplasma infections; infections of Rickettsia and Apicomplexa protozoans were also studied. Of the 270 individually assayed Rhipicephalus haemaphysaloides ticks, A. phagocytophilum was identified in a nymphal tick. Parasites most similar to Anaplasma bovis, Rickettsia rickettsii, Rickettsia sp. TwKM01, and at least seven apicomplexan species (including genera Cryptosporidium, Hepatozoon, and Theileria) were also identified. This study shows that A. phagocytophilum does occur in the hard tick in Taiwan, although whether R. haemaphysaloides can vector this pathogen remains to be determined. This work also reveals a high diversity of tick-borne bacteria and protozoans circulating in a small region and calls for further research on their potential risks for human health.     

A morphological and phylogenetic analysis of <Emphasis Type="Italic">Ornithodoros marinkellei</Emphasis> (Acari: Argasidae), with additional notes on habitat and host usage

Ornithodoros marinkellei was described from larvae collected on Pteronotus spp. bats in Colombia and Panama. More recently, this tick was reported in the Brazilian Amazon. Because some morphometric differences were observed between O. marinkellei larvae from Colombia and Brazil, it was proposed that further investigations were needed to assess whether the differences could be attributed to intra- or inter-specific polymorphism. Herein, we collected O. marinkellei specimens in the type locality of Colombia, in Brazil, and in a new locality in Nicaragua, expanding the distribution of the species to Nicaragua. Morphometric analysis of larvae and adults, corroborated by a principal component analysis (PCA), indicated that the Brazilian specimens were larger than specimens from Colombia and Nicaragua. Phylogenetic analysis inferred from the mitochondrial 16S rRNA gene showed ticks from Colombia and Nicaragua more genetically related than any of them with ticks from Brazil, although ticks from the three countries grouped in a clade sister to a major clade containing sequences of various Neotropical Ornithodoros species. We concluded that ticks identified as O. marinkellei from Colombia, Nicaragua, and Brazil represent the same taxon, and that the genetic and morphological differences between them are likely to have a geographical bias. We redescribed the nymph of O. marinkellei, which has a vestigial hypostome, probably incompatible with blood feeding. We also report human infestation by O. marinkellei adults. As all reports of O. marinkellei adults have been from hot caves (temperature?>?35 °C), this abiotic condition could be a limiting factor for the occurrence of this tick species.     

Co-feeding as a route for transmission of <Emphasis Type="Italic">Rickettsia conorii israelensis</Emphasis> between <Emphasis Type="Italic">Rhipicephalus sanguineus</Emphasis> ticks

The essential oil from Amyris balsamifera (Rutaceae) and elemol, a principal constituent of the essential oil of Osage orange, Maclura pomifera (Moraceae) were evaluated in in vitro and in vivo laboratory bioassays for repellent activity against host-seeking nymphs of the blacklegged tick, Ixodes scapularis, and the lone star tick, Amblyomma americanum. Both bioassays took advantage of the tendency of these host-seeking ticks to climb slender vertical surfaces. In one bioassay, the central portion of a vertical strip of filter paper was treated with test solution and ticks placed or allowed to crawl onto the untreated lower portion. In the other bioassay, a strip of organdy cloth treated with test solution was doubly wrapped (treatment on outer layer) around the middle phalanx of a forefinger and ticks released on the fingertip. Both amyris oil and elemol were repellent to both species of ticks. Elemol did not differ significantly in effectiveness against A. americanum from the widely used repellent deet. At 2 and 4 h after application to filter paper, 827 μg amyris oil/cm² paper repelled 80 and 55%, respectively, of A. americanum nymphs. Ixodes scapularis was repelled by lower concentrations of amyris oil and elemol than A. americanum.     

Detection of <Emphasis Type="Italic">Anaplasmataceae</Emphasis> agents and co-infection with other tick-borne protozoa in dogs and <Emphasis Type="Italic">Rhipicephalus sanguineus</Emphasis> sensu lato ticks

Anaplasmosis and ehrlichiosis are of serious health concern worldwide for animals and humans. In the present study, we report the occurrence of Anaplasma platys and Ehrlichia canis in dogs and Rhipicephalus sanguineus sensu lato (s.l.) ticks from Peninsular Malaysia using a nested polymerase chain reaction assay based on amplification of the 16S rRNA gene. Anaplasma platys was detected from dogs and ticks with prevalence rates of 3.3% (8/240) and 2.9% (4/140), respectively. On the other hand, 12.9% (31/240) of the dogs and 0.7% (1/140) of the ticks were tested positive for E. canis. Additionally, co-infections of A. platys and E. canis with Babesia or Hepatozoon protozoa were also noted in this study. Double infection (E. canis?+?B. gibsoni) was observed in tick, whereas triple infections (E. canis?+?A. platys?+?B. vogeli and E. canis?+?A. platys?+?H. canis) were found in dogs. This study represents the first evidence of A. platys DNA in R. sanguineus s.l. in Malaysia.     

Ecological preferences and seasonal dynamics of ticks (Acari: Ixodidae) on and off bovine hosts in the Eastern Cape Province,South Africa

A total of 31,425 ticks were collected from 10 cattle and also from six drag-samples during the 12-month study period. Adult ticks were removed from the right hand side of each animal and all instars of ticks were placed in containers filled with 70% ethanol. Based on morphological traits, 10 tick species were identified: Rhipicephalus (Boophilus) decoloratus (32.5%), R. evertsi evertsi (18.8%), R. appendiculatus (17.3%), Amblyomma hebraeum (16.3%), R. simus (7.7%), Ixodes pilosus (3.8%), Hyalomma rufipes (3.5%), R. follis (0.08%), Haemaphysalis elliptica (0.04%), H. silacea (0.02%). The southern African yellow dog tick, H. elliptica, was only found on vegetation. The agro-ecological zones differ significantly in tick species and their distribution. The A. hebraeum and R. evertsi evertsi counts were higher in Kowie Thicket (KT) during summer season (2.05 ± 0.01 and 1.00 ± 0.09, respectively) compared to Bedford Dry Grassland (BDG) and Bhisho Thornveld (BT) veld types. In all vegetation types, R. appendiculatus had higher counts in KT in spring (0.91 ± 0.08), summer (0.78 ± 0.08) and winter (0.78 ± 0.08). Rhipicephalus (Boophilus) decoloratus was more frequent in the BT (1.78 ± 0.11) during the summer season. BDG had lower tick infestation with R. evertsi evertsi being the most frequent species in summer. No H. rufipes was collected in the KT. Of epidemiological interest, R. (B.) microplus was absent in the study area which needs further investigation. Within the context of this study, we found agro-ecological differences and seasonal variations to have influence on tick species distribution.