Leachi Group Research Articles

MALDI-TOF MS identification of cattle ticks from Cameroon

MALDI-TOF MS has recently been proposed as an accurate tool for arthropod identification, including ticks. In this study, we evaluate and confirm the ability of MALDI-TOF MS, to identify different tick species collected in Cameroon, considering other lines of evidence (morphology and molecular). A total of 1483 adult ticks were collected from cattle in five distinct sites in the Western Highland of Cameroon. Because of engorged status and/or absence of some morphological criteria, some Ixodes spp. and Rhipicephalus spp. were identified to the genus level only. Among those, 944 ticks (543 males and 401 females) were selected for the current work. They were classified into 5 genera and 11 species: Rhipicephalus (Boophilus) microplus (31.7%), Rhipicephalus lunulatus (26%), Amblyomma variegatum (23%), Rhipicephalus sanguineus s.l. (4.8%), of Haemaphysalis leachi group (4.6%), Hyalomma truncatum (2.6%), Hyalomma rufipes (1.7%), Rhipicephalus muhsamae (1.1%), Rhipicephalus (Boophilus) annulatus (0.6%), Rhipicephalus (Boophilus) decoloratus (0.3%), Ixodes rasus (0.1%), Ixodes spp. (0.2%) and Rhipicephalus spp. (3.3%). Tick legs were subjected to MALDI-TOF MS analyzes, and the spectra of 929 (98.4%) specimens were of good quality. Analysis of these spectra provided intra-species reproducibility and interspecies specificity of MS profiles obtained from the different species. Our in-house MALDI-TOF MS arthropod database was upgraded with spectra from 44 specimens of 10 different tick species. Blind testing of good quality spectra revealed that 99% agreed with the morphological identification. Of these, 96.9% had log score values (LSVs) between 1.73 and 2.57. MALDI-TOF MS also allowed to correct the morphological misidentification of 7 ticks, and to identify 32 engorged ticks that were not morphologically identifiable to the species level. This study supports MALDI-TOF MS as a reliable tool for tick identification and provides new data on tick species identification in Cameroon.

Ticks (Acari: Ixodida) of the genus Haemaphysalis Koch, 1844 in Senegal: a review of host associations, chorology, and identification

The Haemaphysalis genus (Acari, Ixodidae) in Senegal is reviewed. This embodies a summary of specimens collected from vertebrate hosts over three decades. 454 collections were performed over this period (408 from mammals and 46 from birds), representing a total of 5752 ticks in different developmental stages. Seven Haemaphysalis spp. were collected, identified, and inventoried including: H. (Kaiseriana) rugosa, H. (Ornithophysalis) hoodi, H. (Rhipistoma) houyi, as well as four other species belonging to the leachi group, namely H. (Rhipistoma) leachi, H. (Rh.) moreli, H. (Rh.) muhsamae and H. (Rh.) spinulosa. Vertebrate hosts of Haemaphysalis species were identified and listed in different ecological zones of Senegal. An identification key of the haemaphysalids of Senegal is proposed, which is also applicable to the haemaphysalid fauna of the Occidental sub-region of the Afrotropical zoogeographical region. The role of these species as potential vectors of zoonotic diseases in Senegal is also discussed.

In search of the vector(s) of Babesia rossi in Nigeria: molecular detection of B. rossi DNA in Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) ticks collected from dogs, circumstantial evidence worth exploring.

The brown dog tick Rhipicephalus sanguineus (sensu lato) (Acari: Ixodidae) has a cosmopolitan distribution, is a proven vector of a host of pathogens with emerging evidence incriminating it in the transmission of some others. Specifically it is reputed as the main vector of Babesia vogeli whereas the southern African yellow dog tick Haemaphysalis elliptica, long considered to be H. leachi, is apparently the only proven vector of B. rossi, since the resurrection of the separate species H. elliptica as a member of the leachi-group by Apanaskevich et al. However, recent epidemiological surveys conducted in Nigeria show higher prevalence of B. rossi than B. vogeli infection in dogs most of whom were infested with R. sanguineus and rarely with ticks of the H. leachi group. The discrepancy between tick distribution and Babesia spp. prevalent in dogs stimulated us to investigate the possible role of R. sanguineus (s.l.) in the natural transmission of B. rossi. Out of a total of 66 tick samples identified morphologically and molecularly as R. sanguineus collected from dogs manifesting clinical signs of tick-borne diseases, eight (12%) were positive in nested PCR for Babesia sp. DNA. Sequencing results for these amplified products showed that all of the 18S rDNA sequences (693bp) were identical to each other, and bore 99.3-99.9% identities with those from other B. rossi isolates accessible in GenBank. None of the ticks harbored the DNA of B. vogeli or B. canis. The possible implications for the detection of B. rossi DNA in R. sanguineus (s.l.) ticks collected from dogs in the epidemiology of B. rossi infection of dogs in Nigeria is highlighted.

An update on the ecological distribution of the Ixodidae ticks in Zimbabwe.

In total 7657 ticks were collected from 121 dip tanks in 12 districts representative of Zimbabwe's five ecological regions between September 2013 and May 2014. Based on morphological traits four genera and 13 species of ticks were identified. Amblyomma hebraeum (60.3%), Rhipicephalus microplus (58.7%), Rhipicephalus decoloratus (47.1%), Rhipicephalus appendiculatus (56.2%), Rhipicephalus evertsi evertsi (67.8%), Rhipicephalus (near) punctatus (13.2%), Hyalomma truncatum (38%) and Hyalomma rufipes (46.3%) were found in all the ecological regions of the country. Amblyomma variegatum and Rhipicephalus compositus (0.8%) were only found in the north central part of the country while Rhipicephalus simus (5%) had a sparse distribution. The Haemaphysalis leachi group (1.7%) and Rhipicephalus sanguineus (1.7%) were found whenever dogs were sampled suggesting these could be widespread throughout the country. The study confirmed the continued limited distribution of A. variegatum (3.3%) in the north central parts of the country, whereas A. hebraeum was found to have a wide distribution also encroaching areas of high rainfall and lower temperatures where it was not previously recorded. A parapatric relationship existed between these two Amblyomma species. Rhipicephalus appendiculatus was also widely distributed although its presence was dominant in the cooler and wetter parts of the country. The traditionally held view that Hyalomma species and R. evertsi evertsi can survive well under diverse conditions is upheld in this study. Rhipicephalus microplus was also present in dry regions but its adaptability to these regions requires further investigation.

Notes on AfricanHaemaphysalisticks: XVI.H. (Rhipistoma) subterrasp. n., a new member of theleachigroup (Ixodoidea: Ixodidae)

ABSTRACT The female, male, nymph, and larva of Haemaphysalis (Rhipistoma) subterra sp. n. are described. This species is a small-sized member of the H. leachi group with 3/3 dental formula, curving ventral spurs on palpal segment 3, and a moderately punctated scutum. It is recorded from Ethiopia, Kenya, South Africa, Tanzania, Zaire, Zambia, and Zimbabwe. Hosts of adults are carnivores [Herpestes sanguineus (Ruppell), H. ichneumon (Linnaeus), Ichneumia albicauda (Cuvier), Mungos mungo (Gmelin), Atilax paludinosus (Cuvier), Genetta maculata (Gray)], while immatures are collected chiefly from rodents [Aethomys chrysophilus (De Winton), Tachyoryctes splendens Ruppell, Praomys natalensis (Smith)]. One nymph was collected from H. sanguineus, a carnivore, in Zimbabwe.

The Brown Ear Tick Rhipicephalus appendiculatus Neumann (Acarina: Ixodidae) and Associated Tick Species on Wild and Domestic Hosts at Muguga, Kenya

During a study of an experimental population of Rhipicephalus appendiculatus and its transmission of bovine theileriosis to cattle, some supplementary information was obtained on the role of rodents and the small carnivores in the development and maintenance of R. appendiculatus populations. Observations were also made on other common tick species found on these hosts. A total of 530 individual hosts were examined during the 4-yr period, of which the majority included, among the carnivores, the white-tailed mongoose (Ichneumia albicauda), the large spotted genet (Genetta tigrina) and the zorilla (Ictonyx striatus). Among the 482 rodents 95% included Otomys irroratus, Rhabdomys pumilio and Lophuromys flavopunctatus. While all stages of R. appendiculatus thrived on cattle the role of rodents and carnivores was negligible. Similarly, while the other tick species like H. leachi group, immatures of R. hurti/jeanneli group and I. thomasae avoided cattle, they thrived on the carnivores and rodents. However, the adults of R. hurti and R. jeanneli preferred cattle to carnivores and rodents.

Ixodid and Argasid ticks in Connecticut, U.S.A.:Aponomma latum, Amblyomma dissimile, Haemaphysalis leachigroup, andOrnithodoros kelleyi(Acari: Ixodidae, Argasidae)

ABSTRACT Three exotic ticks, Aponomma latum (Koch) engorging on the snake Python regius, Amblyomma dissimile Koch on the boa Epicrates cenchria and Haemaphysalis leachi group feeding on a Rhodesian Ridgeback dog, Canis familiaris, were collected in Connecticut, U.S.A. A single specimen of the bat-feeding tick, Ornithodoros kelleyi Cooley and Kohls, was also collected. The Rhodesian Ridgeback imported from the Republic of South Africa, died from a Babesia canis infection 6 days after entering the United States.

Notes on African Haemaphysalis Ticks. XIV. Description of Adults of H. (Rhipistoma) paraleachi sp. n., a Carnivore Parasite of the H. (R.) leachi Group (Ixodoidea: Ixodidae)

Haemaphysalis (Rhipistoma) paraleachi sp. n. is described from males and females taken from a civet, Viverra civetta (Schreber), in Cameroun. This tick is a large-sized member of the H. (R.) leachi group with a 5/5 or 6/6 dental formula; moderately long, curving ventral spurs of palpal segment 3; and numerous, mostly medium-sized scutal punctations. It is widely distributed in Africa. Adults parasitize small- and medium- sized carnivores. Immatures probably feed chiefly on rodents but cannot be identified with confidence until laboratory-reared samples from known parents are available for study. Haemaphysalis (Rhipistoma) paraleachi sp. n. is described from adults taken from a civet, Vi- verra civetta (Schreber), in Cameroun. A member of the leachi subgroup of the H. (R.) leachi group of African carnivore parasites, this new species is related to H. (R.) leachi (Audouin) 1827, H. (R.) moreli Camicas, Hoogstraal, and El Kam- mah (1972), and H. (R.) punctaleachi Camicas, Hoogstraal, and El Kammah (1973). Data are presented herein only for adult H. (R.) paraleachi parasitizing medium-sized car- nivores in Africa. Immature stages probably feed chiefly on rodents but cannot be identified with confidence until laboratory-reared samples from known parents are available for study.

Seasonal and Regional Abundance of Ticks (Ixodidae) on Hares (Leporidae) in Kenya1

Between August 1967 and July 1968, 367 hares, Lepus capensis , were examined for ticks at 3 ecologically different localities in the Rift Valley of Kenya. Among the 3865 ticks collected, the largest number (2122) and the greatest variety of tick species (9) were from Olorgesailie where the mean annual temperature and rainfall were intermediate between the hotter and drier Magadi and the colder and wetter Akira Ranch. The most frequently collected ticks (totalling more than 100) in these areas were as follows: Magadi— Hyalomma sp. (immatures); Olorgesailie— Hyalomma sp. (immatures), Rhipicephalus pravus group (all stages), R. pulchellus (immatures) and Haemaphysalis leachi group (all stages); and Akira Ranch— Hyalomma sp. (immatures), and Rhipicephalus evertsi evertsi (immatures). A total of 759 ticks was also taken from 30 Lepus crawshayi at localities, 5 in Kenya and 1 in Uganda. Of interest was the collection of 143 larvae, 113 nymphs and a single adult of R. appendiculatus from 12 hares taken at Lolgorien-Kilgoris, a much wetter area than the 3 L. capensis study sites. The percentage of tick-infested hares was related to monthly rainfall, and more adult male hares carried ticks than did female or young hares.

Notes on African Haemaphysalis Ticks. XII. H. (Rhipistoma) zumpti sp. n., a Parasite of Small Carnivores and Squirrels in Southern Africa (Ixodoidea: Ixodidae)

Haeniaphysalis (Rhipistoma) zumpti sp. n., a small, distinctive member of the H. (R.) leachi group, is described from numerous adult and immature samples from a variety of ecological conditions up to 5,000 ft altitude between 33 and 13? S in eastern regions of the Republic of South Africa, Botswana, Zambia, and Malawi. Host records include 10 species in 9 genera: a ground squirrel (Xerus), a bush squirrel (Paraxerus), 4 viverrids (Cynictis, Suricata, Genetta, Herpestes), a mustelid (Ictonyx), a protelid (Proteles), and a felid (Felis). In certain areas, this species appears to be specially adapted to life in burrow systems excavated by ground squirrels and inhabited also by the yellow mongoose and other small carnivores. Here H. (R.) zumpti has optimal opportunities to acquire and transmit pathogens among 2 or more kinds of mammals. Haemaphysalis (Rhipistoma) zumpti sp. n., a distinctive member of the H. (R.) leachi group, is described from adults and immatures from the Republic of South Africa, Botswana, Zambia, and Malawi. In some areas, this species appears to be specially adapted to life in burrow systems excavated by ground squirrels and inhabited also by the yellow mongoose and other small carnivores. From the nature and variety of recorded hosts, more detailed biological and epidemiological investigations of H. (R.) zumpti should be especially rewarding. Haemaphysalis (Rhipistoma) zumpti sp. n. Zumpt's southern Africa haemaphysalid (Figs. 1-35)

Notes on African Haemaphysalis Ticks. VIII. H. (Rhipistoma) moreli sp. n., a Carnivore Parasite of the H. (R.) leachi Group (Ixodoidea: Ixodidae)

Haenmaphysalis (Rhipistoma) moreli sp. n., described from wild-caught adult and laboratory-reared immature samples, is a member of the African H. (R.) leachi group, which also includes H. (R.) leachi (Audouin) and H. (R.) spinulosa Neumann. Rhipistoma ellipticum Koch is a nomen nudum and H. muhsamae Santos Dias and H. ethiopica Santos Dias are junior synonyms of H. (R.) spinulosa Neumann. Records are provided for 701 adult and 126 immature H. (R.) moreli taken from 128 hosts in savannas of western and eastern Africa between about 15? N and 06? S. Hosts of adults are carnivores, chiefly genets and civets, and also the lion, leopard, wildcat, serval, hyena, jackal, fox, and domestic cats and dogs. Immature stages are recorded from rodents and the nest of a bushbaby (Galago senegalensis subsp.). The African Haemaphysalis (Rhipistoma) leachi group appears to have evolved from an Asian progenitor similar to H. (R.) canestrinii (Supino) (Hoogstraal, 1971). The African group has long been the most taxonomically troublesome and biologically confusing assemblage within the genus Haemaphysalis. These problems are of more than academic interest since "H. leachi" has been incriminated in transmission of numerous pathogens causing diseases in man and animals (Neitz, 1956; Hoogstraal, 1967). The incorrect identification of many tick species has often seriously reduced the practical value of literature and of field and laboratory investigations and has caused suggested measures for prevention and control to be questionable. This report is the first in a series in which we hope to increase the understanding of the H. (R.) leachi group Received for publication 9 May 1972. *From Research Project MF12.524.009-3010B, Bureau of Medicine and Surgery, Department of the Navy, Washington, D. C. The opinions and assertions contained herein are the private ones of the authors and are not to be construed as official or as reflecting the views of the Department of the Navy or of the naval service at large. The illustrations in this report were prepared under the auspices of Agreement 03-005-01 between the NIAID (NIH) and NAMRU-3. t Medical entomologist, ORSTOM (Office de la Recherche Scientifique et Technique Outre-Mer), c/o Institut Pasteur, B.P. 220, Dakar, Senegal. t Medical Zoology Department, United States Naval Medical Research Unit Number Three (NAMRU-3), U. S. Interests Section, c/o Spanish Embassy, Cairo, Arab Republic of Egypt. and to stabilize its nomenclature. A species common on carnivores in middle Africa is described. Four taxa have previously been proposed for the H. (R.) leachi group in Africa and another (Rhipistoma ellipticum) has also been referred to this group by certain authors. These are: H. leachi (Audouin, 1827), as redescribed and redefined by Hoogstraal (1958). We consider this to be a valid name for the taxon. H. spinulosa Neumann, 1906, as redescribed and redefined by Hoogstraal (1964). We consider this to be a valid name for the taxon. [Rhipistoma ellipticum Koch, 1844. See Nuttall and Warburton, 1915, p. 517, for discussion of this taxon, which these and earlier authors considered to be a junior synonym of H. leachi (Audouin, 1827). The type material (a single male) appears to have been lost, and we consider R. ellipticuin to be a nomen nudum.] H. muhsamae Santos Dias (1954) was described from one male, one female, and one nymph. Santos Dias (1955) redescribed the male from another specimen, provided new descriptions for the female and nymph, and stated that the original descriptions of the female and nymph were in error and represented a different species (see H. ethiopica). As stated below, we consider this taxon to be a junior synonym of H. spinulosa Neumann, 1906. H. ethiopica Santos Dias (1958) was described from male specimens; the female and nymph of this taxon were stated to be those described under H. muhsamae Santos Dias

Notes on African Haemaphysalis Ticks. IX. H. (Rhipistoma) pedetes sp. n., a Relict Parasite of the Springhare (Rodentia: Pedetidae) in Transvaal (Ixodoidea: Ixodidae)

The male and female of Haemaphysalis (Rhipistoma) pedetes sp. n. are described. This distinctive parasite of the springhare, Pedetes capensis (Pallas) (Rodentia: Pedetidae), in Transvaal appears to represent a relict member of the H. (R.) asiatica group of relatively uncommon species scattered through the Oriental, Palearctic, and Ethiopian Faunal Regions. Factors influencing the wide distribution of numerous tickborne viruses remain unclear (Hoogstraal, 1972) but are obviously associated with historical and ecological factors controlling the distribution of ticks and vertebrate hosts. In this context, knowledge of the presence of relict tick species such as Haemaphysalis (Rhipistoma) pedetes sp. n. may contribute to understanding the history, relationships, and distribution of tickborne viruses. H. (R.) pedetes sp. n. is an easily recognized species appearing to be a member of the H. (R.) asiatica group (Hoogstraal and Morel, 1970) of relatively uncommon species scattered through the Oriental, Palearctic, and Ethiopian Faunal Regions. This tick also shows certain relationships to the H. (R.) erinacei group of the Palearctic Region. It differs distinctly from the well known, more recent H. (R.) leachi group, which is commonly associated as immatures with rodents and as adults with carnivores throughout the Ethiopian Region. The two specimens recorded here were reported by Warburton (1933) and Cooley (1934) as H. numidiana Nuttall, and by Hoogstraal (1955, p. 230, footnote) as an undescribed species. Despite rather intensive Received for publication 9 May 1972. *From Research Project MF12.524.009-3010B, Bureau of Medicine and Surgery, Department of the Navy, Washington, D. C. The opinions and assertions contained herein are the private ones of the author and are not to be construed as official or as reflecting the views of the Department of the Navy or of the naval service at large. The illustrations in this report were prepared under the auspices of Agreement 03-005-1 between the NIAID (NIH) and NAMRU-3. investigation of African ticks by numerous workers during the past two decades, no other samples of this new species have been observed. Therefore we now describe this small sample in the hope of stimulating further study of this exceptionally interesting species and its biology and vector potential. Haemaphysalis (Rhipistoma) pedetes sp. n. South African springhare haemaphysalid (Figs. 1-20)

Studies on Southeast Asian Haemaphysalis Ticks (Ixodoidea, Ixodidae). Redescription of Type Material of H. (Rhipistoma) heinrichi Schulze, a Parasite of the Ferret-Badger in Burma and Vietnam

The male and female of Haemaphysalis (Rhipistoma) heinrichi Schulze, 1939, are redescribed from material in the type series, from a ferret-badger, Melogale p. personata Geoffroy, near Mandalay, Burma. This poorly known species, which was also recorded parasitizing a human, is easily recognized in the Asiatic segment of the H. (R.) leachi group. A male, which differs slightly in chaetotaxy, and a female, both from Melogale personata tonquina Thomas, in the Danang area, Republic of Vietnam, are also reported. Haemaphysalis (Rhipistoma) heinrichi Schulze is redescribed from male and female syntype material, and a lectotype male is designated. The brief original description and crude accompanying illustrations are insufficient for species recognition and for determination of species relationships. H. (R.) heinrichi is now considered to be a member of the carnivoreinfesting H. (R.) leachi group and related to, though clearly distinct from, H. (R.) indica Warburton of the Indian Subregion and from H. (R.) koningsbergeri Warburton and Nuttall of Southeast Asia. The chief host appears to be the ferret-badger, Melogale personata subspp. Schulze (1939) also reported a male from a human. This species is known only within a 30-km radius of Mandalay, Burma, and from near sea level and 450 m elevation in the Danang area, Republic of Vietnam. Haemaphysalis (Rhipistoma) heinrichi Schulze Heinrich's Ferret-Badger Haemaphysalid (Figs. 1-30) Haemaphysalis heinrichi Schulze, 1939, p. Received for publication 10 May 1968. * From Research Project Number MR005.09-0028, Bureau of Medicine and Surgery, Department of the Navy, Washington, D. C. The opinions and assertions contained herein are the private ones of the authors and are not to be construed as official or as reflecting the views of the Department of the Navy or of the naval service at large. t Head, Department of Medical Zoology, United States Naval Medical Research Unit Number Three, Cairo, Egypt, UAR. + Sanitarian Director, United States Department of Health, Education, and Welfare, PHS, NIH, NIAID, Rocky Mountain Laboratory, Hamilton, Montana 59840. 726 to 728, figs. 3, 4; $ and 9 briefly described and illustrated by crude sketches, nymph briefly described; holotype $ from B rmese Ferret-Badger, Melogale (= Helictis) personata personata Geoffroy, Popa, 600 m altitude Burma, G. Heinrich legit. Schulze stated that the holotype was deposited in the Berlin Zoological Museum but, during a recent visit to this institution, Dr. Harold Trapido (pers. comm., 1965) failed to find specimens labeled H. heinrichi. Schulze also noted a $ taken on a human (together with 19 Dermacentor auratus) at Lamaing, but did not specify the number(s) or disposition of male, female, or nymphal specimens in his possession. The Schulze collection in the Rocky Mountain Laboratory contains 3 $ & and 19 of H. heinrichi with a Schulze label reading Helictis personatus de Ber. Z. M. 147. Popa, 600 m, Burma, leg. G. Heinrich. Thus, the material described herein is apparently from the type series and presumably includes the intended holotype specimen. One $ in good condition is hereby designated as the Lectotype and illustrated (Figs. 1, 2, 5-12); and 2 other $ $ are in fair or poor condition; the 9 is engorged, in fair condition. The same vial also contains 1 Haemaphysalis wellingtoni Nuttall and Warburton and 2 nymphs and 7 larvae of Dermacentor sp. The present whereabouts of the H. heinrichi nymph mentioned by Schulze is unknown.