Historical dispersal and host-switching formed the evolutionary history of a globally distributed multi-host parasite - The Ligula intestinalis species complex.

Studies on parasite biogeography and host spectrum provide insights into the processes driving parasite diversification. Global geographical distribution and a multi-host spectrum make the tapeworm Ligula intestinalis a promising model for studying both the vicariant and ecological modes of speciation in parasites. To understand the relative importance of host association and biogeography in the evolutionary history of this tapeworm, we analysed mtDNA and reduced-represented genomic SNP data for a total of 139 specimens collected from 18 fish-host genera across a distribution range representing 21 countries. Our results strongly supported the existence of at least 10 evolutionary lineages and estimated the deepest divergence at approximately 4.99-5.05 Mya, which is much younger than the diversification of the fish host genera and orders. Historical biogeography analyses revealed that the ancestor of the parasite diversified following multiple vicariance events and was widespread throughout the Palearctic, Afrotropical, and Nearctic between the late Miocene and early Pliocene. Cyprinoids were inferred as the ancestral hosts for the parasite. Later, from the late Pliocene to Pleistocene, new lineages emerged following a series of biogeographic dispersal and host-switching events. Although only a few of the current Ligula lineages show narrow host-specificity (to a single host genus), almost no host genera, even those that live in sympatry, overlapped between different Ligula lineages. Our analyses uncovered the impact of historical distribution shifts on host switching and the evolution of host specificity without parallel host-parasite co-speciation. Historical biogeography reconstructions also found that the parasite colonized several areas (Afrotropical and Australasian) much earlier than was suggested by only recent faunistic data.

Ascaris lumbricoides and ticks associated with sensitization to galactose α1,3-galactose and elicitation of the alpha-gal syndrome.

BACKGROUND: IgE to galactose alpha-1,3 galactose (alpha-gal) causes alpha-gal syndrome (delayed anaphylaxis after ingestion of mammalian meat). Development of sensitization has been attributed to tick bites; however, the possible role of other parasites has not been well studied. OBJECTIVE: Our aims were to assess the presence, relative abundances, and site of localization of alpha-gal-containing proteins in common ectoparasites and endoparasites endemic in an area of high prevalence of alpha-gal syndrome, as well as to investigate the ability of ascaris antigens to elicit a reaction in a humanized rat basophil in vitro sensitization model. METHODS: Levels of total IgE, Ascaris-specific IgE, and alpha-gal IgE were measured in sera from patients with challenge-proven alpha-gal syndrome and from controls without allergy. The presence, concentration, and localization of alpha-gal in parasites were assessed by ELISA, Western blotting, and immunohistochemistry. The ability of Ascaris lumbricoides antigen to elicit IgE-dependent reactivity was demonstrated by using the RS-ATL8 basophil reporter system. RESULTS: Alpha-gal IgE level correlated with A lumbricoides-specific IgE level. Alpha-gal protein at 70 to 130 kDa was detected in A lumbricoides at concentrations higher than those found in Rhipicephalus evertsi and Amblyomma hebraeum ticks. Immunohistochemistry was used to localize alpha-gal in tick salivary acini and the helminth gut. Non-alpha-gal-containing A lumbricoides antigens activated RS-ATL8 basophils primed with serum from subjects with alpha-gal syndrome. CONCLUSION: We demonstrated the presence, relative abundances, and site of localization of alpha-gal-containing proteins in parasites. The activation of RS-ATL8 IgE reporter cells primed with serum from subjects with alpha-gal syndrome on exposure to non-alpha-gal-containing A lumbricoides proteins indicates a possible role of exposure to A lumbricoides in alpha-gal sensitization and clinical reactivity.

28S rRNA sequences for Linguatula spp.

Identification of specimens belonging to the genus Linguatula (Pentastomida) is relatively easy due to their unique morphology. However, differentiation between species of Linguatula can be challenging for several reasons, including considerable differences between different developmental stages of the parasite within and between species. Currently, 18S rRNA and Cox1 sequences are the only available comparable sequences in GenBank, but recent research has discussed the utility of 28S rRNA for pentastomid phylogenetics. This study presents 28S rRNA gene sequences for two members of the genus Linguatula. Sequences of 28S rRNA were successfully obtained from well-identified samples of L. serrata (collected in Australia) and L. nuttalli (collected in South Africa), with voucher specimens. Phylogenetic analysis of the 28S rRNA region showed 6% difference between L. serrata and L. nuttalli, with low levels of intraspecific variation. In comparison, 18S rRNA and Cox1 sequences from the same specimens showed 0.23% and 13% interspecific differences, respectively. The results of this study show that 28S rRNA has greater genetic diversity to allow for improved differentiation between species of Linguatula than 18S rRNA but is on par with Cox1. Records that do not provide adequate morphological or molecular data to justify independent specific diagnoses must be regarded cautiously, and the need for continued research on species of Linguatula, using a combined morphological and molecular analysis, across a number of different hosts, development stages, geographical regions and molecular markers is highlighted.

A new species of Clinostomum Leidy, 1856 based on molecular and morphological analysis of metacercariae from African siluriform fishes.

In the Afrotropic region, the genus Clinostomum is represented by four accepted and four unnamed species distinguished using molecular data. Here, we describe one of the four unnamed species as Clinostomum ukolii n. sp. based on metacercariae from siluriform fishes (Synodontis batensoda, Schilbe intermedius) collected in Nigeria and South Africa. The new species is distinguished by molecular data (39 new sequences of partial cytochrome c oxidase I ≥ 6.7% divergent from those of other species) and morphological differences from named and unnamed species in the same region. Metacercariae of C. ukolii n. sp. can be distinguished based on size, tegumental spines, and various aspects of the genital complex, including its position, lobation of the anterior testis, and the disposition and shape of the cirrus pouch. Although descriptions of new species of digeneans are typically based on the morphology of adults, we argue that in cases where data are available from metacercariae from regionally known species, new species can be described based on metacercariae, particularly when supported by molecular data, as here. Moreover, sub-adult reproductive structures can be clearly visualized in metacercaria of Clinostomum. Considering metacercariae as potential types for new species could advance clinostome systematics more rapidly, because metacercariae are encountered much more often than adults in avian definitive hosts.

Phylogeny and systematics of the Proterodiplostomidae Dubois, 1936 (Digenea: Diplostomoidea) reflect the complex evolutionary history of the ancient digenean group.

The Proterodiplostomidae Dubois, 1936 is a relatively small family of diplostomoidean digeneans parasitising the intestines of reptilian hosts associated with freshwater environments in tropical and subtropical regions. The greatest diversity of proterodiplostomids is found in crocodilians, although some parasitise snakes and turtles. According to the most recent revision, the Proterodiplostomidae included 17 genera within 5 subfamilies. Despite the complex taxonomic structure of the family, availability of testable morphology-based phylogenetic hypotheses and ancient hosts, molecular phylogenetic analyses of the group were practically lacking. Herein, we use novel DNA sequence data of the nuclear lsrRNA gene and mitochondrial cox1 gene from a broad range of proterodiplostomid taxa obtained from crocodilian, fish, and snake hosts on four continents to test the monophyly of the family and evaluate the present morphology-based classification system of the Proterodiplostomidae in comparison with the molecular phylogeny. This first detailed phylogeny for the Proterodiplostomidae challenges the current systematic framework. Combination of molecular phylogenetic data with examination of freshly collected quality specimens and re-evaluation of morphological criteria resulted in a number of systematic and nomenclatural changes along with a new phylogeny-based classification of the Proterodiplostomidae. As the result of our molecular and morphological analyses: (i) the current subfamily structure of the Proterodiplostomidae is abolished; (ii) three new genera, Paraproterodiplostomum n. g., Neocrocodilicola n. g. and Proteroduboisia n. g., are described and Pseudoneodiplostomoides Yamaguti, 1954 is restored and elevated from subgenus to genus level; (iii) two new species, Paraproterodiplostomum currani n. g., n. sp. and Archaeodiplostomum overstreeti n. sp., are described from the American alligator in Mississippi, USA. Comparison of the structure of terminal ducts of the reproductive system in all proterodiplostomid genera did not support the use of these structures for differentiation among subfamilies (or major clades) within the family, although they proved to be useful for distinguishing among genera and species. Our study includes the first report of proterodiplostomids from Australia and the first evidence of a snake acting as a paratenic host for a proterodiplostomid. A key to proterodiplostomid genera is provided. Questions of proterodiplostomid-host associations parasitic in crocodilians are discussed in connection with their historical biogeography. Our molecular phylogeny of the Proterodiplostomidae closely matches the current molecular phylogeny of crocodilians. Directions for future studies of the Proterodiplostomidae are outlined.

Molecular data show Clinostomoides Dollfus, 1950 is a junior synonym of Clinostomum Leidy, 1856, with redescription of metacercariae of Clinostomum brieni n. comb.

The genus Clinostomoides Dollfus, 1950 was erected to accommodate a single worm from Ardea goliath sampled in the Belgian Congo. The specimen was distinguished from other clinostomids by its large size and posterior genitalia. In the following years, metacercariae of Clinostomoides brieni, have been described in Clarias spp. in southern and western Africa. A few authors have referred to Clinostomum brieni, but all such usages appear to be lapsus calami, and the validity of Clinostomoides remains widely accepted. In this study our aim was: position C. brieni among the growing clinostomids molecular database, and redescribe the species with emphasis on characters that have emerged as important in recent work. We sequenced two nuclear (partial 18S and ITS) and one mitochondrial marker (partial cytochrome c oxidase I) and studied morphology in metacercariae from hosts and localities likely to harbour the type species (Clarias spp., Democratic Republic of the Congo, South Africa). Phylogenetic analysis shows C. brieni belongs within Clinostomum Leidy, 1856. We therefore transfer C. brieni to Clinostomum, amend the diagnosis for the genus Clinostomum and provide a critical analysis of other species in Clinostomoides, all of which we consider species inquirendae, as they rest on comparisons of different developmental stages.

Variations in the microbiome due to storage preservatives are not large enough to obscure variations due to factors such as host population, host species, body site, and captivity.

The study of the primate microbiome is critical in understanding the role of the microbial community in the host organism. To be able to isolate the main factors responsible for the differences observed in microbiomes within and between individuals, confounding factors due to technical variations need to be removed. To determine whether alterations due to preservatives outweigh differences due to factors such as host population, host species, body site, and habitat, we tested three methods (no preservative, 96% ethanol, and RNAlater) for preserving wild chimpanzee (fecal), wild lemur (fecal), wild vervet monkey (rectal, oral, nasal, otic, vaginal, and penile), and captive vervet monkey (rectal) samples. All samples were stored below - 20°C (short term) at the end of the field day and then at - 80°C until DNA extraction. Using 16S rRNA gene sequencing, we show a significant preservative effect on microbiota composition and diversity. Samples stored in ethanol and RNAlater appear to be less different compared with samples not stored in any preservative (none). Our differential analysis revealed significantly higher amounts of Enterococcaceae and Family XI in no preservative samples, Prevotellaceae and Spirochaetaceae in ethanol and RNAlater preserved samples, Oligosphaeraceae in ethanol-preserved samples, and Defluviitaleaceae in RNAlater preserved samples. While these preservative effects on the microbiome are not large enough to remove or outweigh the differences arising from biological factors (e.g., host species, body site, and habitat differences) they may promote misleading interpretations if they have large enough effect sizes compared to the biological factors (e.g., host population).

Diversity of haemoprotozoan parasites infecting the wildlife of South Africa.

Tissue samples from wildlife from South Africa were opportunistically collected and screened for haemoprotozoan parasites using nonspecific PCR primers. Samples of 127 individuals were tested, comprising over 50 different species. Haemogregarines were the most commonly identified parasites, but sarcocystids and piroplasmids were also detected. Phylogenetic analyses estimated from the 18S rDNA marker highlighted the occurrence of several novel parasite forms and the detection of parasites in novel hosts. Phylogenetic relationships, which have been recently reviewed, appear to be much more complex than previously considered. Our study highlights the high diversity of parasites circulating in wildlife in this biodiverse region, and the need for further studies to resolve taxonomic issues.

Morphological Studies of Developmental Stages of Oculotrema hippopotami (Monogenea: Polystomatidae) Infecting the Eye of Hippopotamus amphibius (Mammalia: Hippopotamidae) Using SEM and EDXA with Notes on Histopathology.

The present study was performed to observe histopathological effects of Oculotrema hippopotami Stunkard, 1924 infection in the eye of Hippopotamus amphibius, as well as to reveal new details of morphology and structural features of this monogenean and its comparison between 2 age stages of the parasite. This was done using both light and scanning electron microscopy, energy dispersive X-ray analysis (EDXA) and histopathology. The presence of a mixture of different generations (adult and sub-adult) in one host individual is common for Oculotrema Stunkard, 1924 in contrast to Polystoma Zeder, 1800. New metrical and graphical information obtained for adults and sub-adults compared with the previous studies. Here we show the presence of genital papillae in adults, metrical data on the distal part of the vas deferens. SEM micrographs of sperm ejaculatory structures and information about the flattened dorsal side of the body provided for the first time. Histopathological changes, such as necrosis and hemorrhage in host tissues as a result of O. hippopotami attachment structures are described. Structural analysis of different body parts of O. hippopotami of both age groups are also included. We show qualitative differences in the presence of hardening ions (S, P, Ca) in attachment structures (oral and haptor suckers) that increase with the age of the worm. The presence of sub-adults and adults on the same host, together with high levels of infection without high pathogenicity may account for Oculotrema being one of the most successful parasites among the Monogenea.

Redescription of Maupasina weissi (Seurat, 1913) (Nematoda: Ascaridida) from sengis, Elephantulus spp. and Macroscelides proboscideus (Shaw) (Macroscelidea), in Africa.

Maupasina weissi (Seurat, 1913), is redescribed from the eastern rock sengi, Elephantulus myurus Thomas & Schwann, from Limpopo Province and compared with material collected from Free State and North West Provinces, South Africa, as well as the description of the type-material from South Tunisia. Distinguishing features of the species include a corona radiata of 12 pointed leaflike elements, a complex bipartite buccal capsule with 3 large bicuspid denticular lobes and 4 smaller club-shaped lobes and 11 (occasionally 10) pairs of cloacal papillae in the male. The complex taxonomic history of the genus Maupasina Seurat, 1917 is discussed. The probable misidentification of Macroscelides proboscideus Shaw, the short eared sengi, from North West Province, South Africa, as a host of M. weissi is explained, indicating that M. weissi occurs only in species of the genus Elephantulus Thomas & Schwann. The widely separated geographical regions, stretching the length of the African continent, in which M. weissi has been found are indicative of a conservative species with a broad geographical distribution. Tenebrionid beetles, up to 50% of stomach contents of E. myurus in Limpopo Province may act as intermediate hosts in the life-cycle of M. weissi.

Expanded description of Lamproglena cleopatra Humes, 1957 (Lernaeidae: Copepoda) from Labeo spp. (Cyprinidae) with a key to species of Lamproglena von Nordmann, 1832.

The occurrence of the copepod Lamproglena cleopatra Humes, 1957, parasitising freshwater fishes in the Limpopo River System is presented, along with new morphological data. This crustacean was originally described parasitising a cyprinid (Labeo forskalii Rüppell) from the River Nile, Egypt. During 2014-2015 crustacean samples were collected from the gills of three cyprinid fish species, Labeo rosae Steindachner from Flag Boshielo Dam, Labeo molybdinus Du Plessis from Nwanedi-Luphephe Dam in South Africa, and Labeo ruddi Boulenger from the River Bubye in Zimbabwe. The specimens from the present study were morphologically similar regardless of the host, but exhibited some morphometric intraspecific differences in comparison with the type-specimen from Egypt. A description of L. cleopatra copepodid III stage and a taxonomic key to Lamproglena spp. is provided.

An annotated list and molecular data on larvae of gryporhynchid tapeworms (Cestoda: Cyclophyllidea) from freshwater fishes in Africa.

An annotated list of larvae (metacestodes) of gryporhynchid tapeworms (Cestoda: Cyclophyllidea) from freshwater fishes in Africa is provided with numerous new host and geographical records. Newly collected materials from Burundi, Democratic Republic of the Congo, Kenya, Madagascar, Namibia, Senegal, South Africa, Sudan and Zimbabwe practically double the total number of species reported from African fish so far. We confirm the occurrence of 16 species (five unidentified to the species level and most likely representing new taxa) belonging to the genera Amirthalingamia Bray, 1974 (1 species), Cyclustera Fuhrmann, 1901 (2 species), Dendrouterina Fuhrmann, 1912 (1 species), Neogryporhynchus Baer & Bona, 1960 (1 species), Paradilepis Hsü, 1935 (4 species), Parvitaenia Burt, 1940 (5 species), and Valipora Linton, 1927 (2 species). Additionally, metacestodes of four unidentified species of Paradilepis and Parvitaenia are reported from fish for the first time. Rostellar hooks of all species are illustrated and their measurements are provided together with a host-parasite list. The molecular phylogenetic analysis based on partial LSU rDNA sequences offers the first insight into the internal phylogenetic relationships within the family. Together with the morphological observations, the present study provides a taxonomic baseline for future studies on this largely neglected, but widely distributed and relatively frequent, group of parasites of African fishes, including economically important cichlids like tilapias and cyprinids.

A morphological and molecular study of Clinostomid metacercariae from African fish with a redescription of Clinostomum tilapiae.

The genus Clinostomum Leidy, 1856 (Digenea: Clinostomidae) has been reported in all ecozones of the world and a clear separation between the species of the 'Old World' and 'New World' has been recognized based on molecular studies. Recent works on Afrotropical species include redescriptions of C. cutaneum and C. phalacrocoracis, while C. tilapiae has yet to be studied using modern taxonomic approaches. In the present research, morphological redescription of C. tilapiae metacercariae from a new host, Synodontis batensoda sampled at Anambra River Basin, Nigeria, together with molecular analysis of nuclear internal transcribed spacer rDNA and cytochrome c oxidase 1 mtDNA are reported. We also provide morphological and molecular data from four further putative species of Clinostomum (morphotypes 1-4) from different areas of Africa, as well as the first report of C. phalacrocoracis in South Africa.

Guppies (Poecilia reticulata) introducing an alien parasite, Camallanus cotti (Nematoda: Camallanidae) to Africa, the first report.

Introduced alien fish species and their associated parasites may result in a serious threat to indigenous biodiversity. Furthermore, this may have negative impacts on cultured fish as well as on native parasitic fauna. In the present study, the invasive Asian nematode, Camallanus cotti Fujita, 1927 (Nematoda: Camallanidae), is reported from the guppy (Poecilia reticulata) for the first time in Africa. This parasite is assumed to be introduced into Africa along with the introduction of exotic poeciliid fishes, which are known to be the most common hosts of C. cotti in ornamental fish industry worldwide.The presence of this parasite in both aquarium-cultured fish as well as fish from natural waterbodies is evidence of the introduction of the alien organisms due to insufficient prophylactic veterinary control during transfer of non-native hosts between countries and the spread of them by the anthropogenic introduction to natural systems.

Description and phylogenetic position of a new species of Rhabdias Stiles et Hassall, 1905 (Nematoda: Rhabdiasidae) from the banded rubber frog, Phrynomantis bifasciatus (Smith) (Amphibia: Microhylidae), in South Africa.

The lung-dwelling nematode Rhabdias engelbrechti sp. n. was found in five of eight examined banded rubber frogs in Limpopo Province, South Africa. The species is differentiated from species of Rhabdias Stiles et Hassall, 1905 occurring in the Afrotropical Realm based on the presence of a globular cuticular inflation at the anterior end, the buccal capsule walls being distinctly divided into anterior and posterior parts, the buccal capsule size (6-9 µm × 16-18 µm), and the body length (3.8-6.1 mm). Rhabdias engelbrechti is the tenth species of the genus found in Afrotropical anurans. Our molecular phylogenetic analysis based on the complete sequences of the ITS region and partial sequences of large subunit (28S) gene of the nuclear ribosomal RNA demonstrates that the new species is more closely related to the Eurasian species Rhabdias bufonis (Schrank, 1788) than to two other species from sub-Saharan Africa represented in the tree. In addition, partial sequences of the mitochondrial protein coding cox1 and ribosomal 12S genes of the new species have shown significant differences from all previously published sequences of these genes from African species of Rhabdias.

A new species of Centrorhynchus Lühe, 1911 (Acanthocephala: Centrorhynchidae) from the lizard buzzard Kaupifalco monogrammicus (Temminck) (Aves: Accipitridae) in South Africa.

Centrorhynchus sarehae n. sp. is described from the lizard buzzard Kaupifalco monogrammicus (Temminik) in Louis Trichardt City, Limpopo Province, South Africa. The new species can be distinguished from all other species of Centrorhynchus Lühe, 1911 except C. gendrei (Golvan, 1957) and C. mariauxi Smales, 2011 in having a dilated region in the posterior trunk of the female. Centrorhynchus sarehae differs from both these species in the characters of the proboscis armature, particularly the number of hooks per row and the lengths of the longest hooks. This is the first record of a fully identified species of Centrorhynchus in South Africa.

The invasive fish tapeworm Atractolytocestus huronensis (Cestoda), a parasite of carp, colonises Africa.

Biological invasions represent a serious threat for aquaculture because many of introduced parasites may negatively affect the health state of feral and cultured fish. In the present account, the invasive tapeworm Atractolytocestus huronensis Anthony, 1958 (Cestoda: Caryophyllidea), which was originally described from North America and has been introduced to Europe including the British Isles with its specific host, common carp (Cyprinus carpio L.), is reported from Africa for the first time. Its recent introduction to South Africa, where it was found in four localities where common carp is cultured, is another evidence of insufficient prophylactic measures and inadequate veterinary control during transfers of cultured fish, especially common carp, between continents. Together with the Asian fish tapeworm, Bothriocephalus acheilognathi, A. huronensis is another fish tapeworm with ability to spread throughout the globe as a result of man-made introductions of its fish hosts.

Phylogenetic affinities and systematic position of Entomelas sylvestris Baker, 1982 (Nematoda: Rhabdiasidae), a parasite of Breviceps sylvestris FitzSimons (Amphibia: Brevicipitidae) in South Africa.

The genus Entomelas Travassos, 1930 currently includes nine species of rhabdiasid nematodes, eight of them parasitic in lizards and only one, Entomelas sylvestris Baker, 1982, parasitic in amphibians. Entomelas sylvestris was originally described from the Forest Rain Frog Breviceps sylvestris FitzSimons in South Africa and was not reported since. It was placed in the genus Entomelas without any specific arguments for this taxonomic decision, presumably mainly based on details of the buccal capsule morphology. We have found this species in the same host in Limpopo province, South Africa. Molecular phylogenetic analysis based on the newly-obtained sequence of complete ITS region and partial nuclear large ribosomal subunit (28S) gene of E. sylvestris and previously published sequences of a variety of other rhabdiasid taxa, has convincingly demonstrated that this species does not belong in Entomelas. Instead, it clustered together with the members of Rhabdias Stiles & Hassall, 1905 from amphibian hosts. Therefore, we transfer E. sylvestris into Rhabdias as Rhabdias sylvestris (Baker, 1982) n. comb. In our analysis E. sylvestris appears, albeit with weak support, as a basal/sister taxon to the rest of Rhabdias spp. which explains to some extent the differences in the buccal capsule morphology between this species and other Rhabdias spp.

First detection of Toxoplasma gondii Africa 4 lineage in a population of carnivores from South Africa.

Introduction: There have only been a few molecular studies conducted on the detection of T. gondii in tissues of carnivores in South Africa, with no data on the genetic diversity of this parasite. That is why the aim of this study was to detect and genotype T. gondii DNA in tissues of selected wild and domestic carnivores in South Africa. Methods: Samples were collected from 80 animals of 20 species (mainly road-killed) in the four provinces of Limpopo (n=57), Mpumalanga (n=21), Gauteng (n=1) and Free State (n=1) during the period 2014-2018. Samples of brain (n=31), heart (n=4), liver (n=40), spleen (n=2) and lung (n=3) were used to detect T. gondii by real-time PCR targeting a 529 bp repeating fragment of T. gondii DNA. Samples that were positive in real-time PCR were genotyped using 15 microsatellite markers. Results: T. gondii DNA was detected in 4 (5 %) samples: in the brain from a Black-backed Jackal (Canis mesomelas), in the liver from a African Wildcat (Felis silvestris lybica) and in the liver and heart of two Rusty-spotted Genets (Genetta maculata) respectively. The DNA sample from Black-backed Jackal was genotyped and characterized as belonging to the type Africa 4 lineage (equivalent to RFLP genotype ToxoDB#20), that is a widespread lineage in Africa. Discussion: This is the first genetic characterization of T. gondii isolated from a wild carnivore on the African continent and the first report of T. gondii in Black-backed Jackal. The Africa 4 lineage was also confirmed in the region of Southern Africa for the first time.

Hard ticks (Acari: Ixodidae) parasitizing bushbabies (Mammalia: Galagidae) in a biodiversity hotspot of northern South Africa.

South Africa has six species of primates, three of which are bushbabies (family Galagidae). Very little information is available on their parasites due to the lack of longitudinal studies, although Rhipicephalus appendiculatus, Amblyomma hebraeum and Haemaphysalis elliptica were previously reported from the brown greater galago (Otolemur crassicaudatus) in South Africa. During 2014-2019, 83 O. crassicaudatus (70 live-trapped and 13 deceased animals) were checked for the presence of hard ticks, all from Limpopo Province, South Africa. Seventy-three of 83 (88 %) galagos were found to be tick-infested. Among ixodid genera, Haemaphysalis had the highest prevalence (46 % of the bushbabies), followed by Rhipicephalus (25 %) and Ixodes (18 %). In total, ten tick species were identified. Importantly, all infestations were monospecific. Ticks occurred on various body parts of bushbabies, thus no predilection site was noted. In conclusion, while previously only three ixodid species were known to infest bushbabies in South Africa, the present study showed that these animals can be parasitized by a much broader range of hard ticks.