James E. Keirans

Analysis of the systematic relationships among ticks of the genera Rhipicephalus and Boophilus (Acari: Ixodidae) based on mitochondrial 12S ribosomal DNA gene sequences and morphological characters

A portion of mitochondrial 12S rDNA sequences (337–355 base pairs) and 63 morphological characters of 36 hard-tick species belonging to 7 genera were analyzed to determine the phylogenetic relationships among groups and species of Rhipicephalus and between the genera Rhipicephalus and Boophilus. Molecular and morphological data sets were first examined separately. The molecular data were analyzed by maximum parsimony (MP), maximum likelihood, and neighbor-joining distance methods; the morphological data were analyzed by MP. After their level of congruence was evaluated by a partition homogeneity test, all characters were combined and analyzed by MP. The branches of the tree obtained by combining the data sets were better resolved than those of the trees inferred from the separate analyses. Boophilus is monophyletic and arose within Rhipicephalus. Boophilus species clustered with species of the Rhipicephalus evertsi group. Most of the clustering within Rhipicephalus was, however, consistent with previous classifications based on morphological data. Morphological characters were traced on the molecular reconstruction in order to identify characters diagnostic for monophyletic clades. Within the Rhipicephalus sanguineus complex, the sequences of specimens morphologically identified as Rhipicephalus turanicus were characterized by a high level of variability, indicating that R. turanicus-like morphology may cover a spectrum of distinct species.

MOLECULAR PHYLOGENETIC ANALYSES INDICATE THAT THE IXODES RICINUS COMPLEX IS A PARAPHYLETIC GROUP

The Ixodes ricinus species complex is a group of ticks distributed in almost all geographic regions of the world. Lyme borreliosis spirochetes are primarily transmitted by tick species within this complex. It has been hypothesized that the Lyme vector ticks around the world are closely related and represent a monophyletic group. This implies that vector competence in ixodid ticks for Lyme agents might have evolved only once. To test this hypothesis, we used a molecular phylogenetic approach. Two fragments of mitochondrial 16S ribosomal deoxyribonucleic acid were sequenced from 11 species in the I. ricinus complex and from 16 other species of Ixodes. Phylogenetic analysis using Bayesian methodology indicated that the I. ricinus complex is not a monophyletic group unless 3 additional Ixodes species are included in it. The known major vectors of Lyme disease agents in different areas of the world are not sister taxa. This suggests that acquisition of the ability to transmit borreliosis agents in species of Ixodes may have multiple origins.

First records of Amblyomma americanum, Ixodes (Ixodes) dentatus, and Ixodes (Ceratixodes) uriae (Acari: Ixodidae) from Maine

The first records of 3 ixodid tick species collected in the state of Maine are reported. A total of 23 records of the lone star tick, Amblyomma americanum (L., 1758), in 11 counties from hosts with no history of travel outside the state demonstrates that this tick is now a resident of Maine. Ixodes dentatus Marx, 1899 is recorded from Waldo and Lincoln counties, and Ixodes uriae White, 1852 is recorded from Matinicus Rock in Knox County. This is the first report of I. uriae from the eastern United States. Disease agents such as those causing human monocytic ehrlichiosis, Lyme disease, Rocky Mountain spotted fever, and several arboviruses have been recorded from 1 or more of these tick species.

Clarification of the Rhipicephalus sanguineus group (Acari, Ixodoidea, Ixodidae). II. R. sanguineus (Latreille, 1806) and related species

AbstractIn this, our second study on the biosystematics of the Rhipicephalus sanguineus group, we consider an additional five species found in the Afrotropical region, namely R. sanguineus, R. camicasi, R. guilhoni, R. moucheti and R. bergeoni.Cross-breeding experiments have shown that R. sanguineus (Latreille, 1806) sensu stricto is genetically distinct from R. camicasi Morel, Mouchet & Rodhain, 1976. Further cross-breeding trials confirmed that R. camicasi also occurs in Saudi Arabia: a field strain from this country was more productive than a laboratory-reared strain from Egypt. R. sanguineus, which parasitises mainly dogs, probably occurs throughout the Afrotropical region. R. camicasi parasitises both large and small domestic ruminants and is restricted to the arid and semi-arid lowlands of northeastern Africa and the Arabian peninsula. The larva, nymph, male and female of these two last named species are described with the aid of scanning electron micrographs (SEMs).The two western African species, R. guilhoni Morel & Vassiliades, 1963 and R. moucheti Morel, 1964, are accepted as valid members of the R. sanguineus group, but we have little material available for critical study. The male and female of R. guilhoni are briefly redescribed with the aid of SEMs. R. bergeoni Morel & Balis, 1976 occurs predominantly on cattle in the wetter highlands and subhighlands of Ethiopia and Sudan. The male and female are briefly described. Current morphological and ecological data suggest that the inclusion of this species in the R. sanguineus group is questionable.

Clarification of the Rhipicephalus sanguineus group (Acari, Ixodoidea, Ixodidae). I. R. sulcatus Neumann, 1908 and R. turanicus Pomerantsev, 1936

AbstractFor many years the biosystematic status of several species in the Rhipicephalus sanguineus group has been confused, with the result that they have often been misidentified. Over the years some 18 of these species have been synonymised with R. sanguineus (Latreille, 1806) itself. More recently four new species in the group have been described: R. guilhoni Morel & Vassiliades, 1963; R. moucheti Morel, 1964; R. bergeoni Morel & Balis, 1976; and R. camicasi Morel, Mouchet & Rodhain, 1976.The literature on this group is critically reviewed and the main systematic problems analysed. R. sulcatus Neumann, 1908 and R. turanicus Pomerantsev, 1936 are two species that were confused both morphologically and ecologically. This has resulted in erroneous conclusions regarding their host relationships and distributions. Integrated biological, morphological and ecological studies on these two species have been conducted. Cross-breeding experiments have proved that both are distinct taxonomic entities. Interbreedings between African and Cypriot strains of R. turanicus demonstrated marked heterosis.A scanning electron microscope was used to determine the main diagnostic morphological differences between the larvae, nymphae and adults of R. sulcatus and R. turanicus, and the morphological similarities between African and Cypriot strains of R. turanicus. Rhipicephalus sulcatus occurs widely in the Afrotropical region in wetter ecological habitats and most frequently parasitises hares, dogs and jackals. R. turanicus occurs more-or-less throughout the Afrotropical region in a wide range of climatic biotopes, as well as in parts of southern Europe, Arabia and Asia, and is most abundant in the late rainy/early dry seasons. It occurs on a wide range of domesticated and wildlife hosts, including ground-feeding birds.

Invasion: Exotic Ticks (Acari: Argasidae, Ixodidae) Imported into the United States. A Review and New Records

Abstract A review of the literature and unpublished records from the U.S. National Tick Collection on the importation of ticks from foreign lands reveals that at least 99 exotic tick species assignable to 11 genera have been either detected and destroyed at ports of entry or inadvertently imported into the United States in the past half century. This number includes four argasid and 95 ixodid species, some of which are important vectors of agents that cause disease to both man and animals. If one includes Aponomma sp. and Hyalomma sp. and the subspecies of Rhipicephalus, the total exceeds 100 taxa. It is notable that the number of imported tick species recorded herein exceeds the total number of tick species native to the United States. It appears that the soft tick genera Argas, Antricola and Nothoaspis have not been imported, although at some point in time Argas persicus (Oken) was introduced because it is resident although not often collected. The hard tick genera Anomalohimalaya, Cosmiomma, Margaropus, Nosomma and Rhipicentor, and the nuttalliellid genus Nuttalliella have also not been imported.

Establishment of the foreign parthenogenetic tick Amblyomma rotundatum (acari : ixodidae) in Florida

The parthenogenetic tick Amblyomma rotundatum, a Central and South American species, has become established in southern Florida. The date of introduction is unknown, but it is suspected to be either during the 1930s, when 1 of its natural hosts, the giant or marine toad, Bufo marinus, was introduced to southern Florida as a potential biological control of pest beetles in sugar cane fields, or between 1955 and 1964 when specimens of B. marinus were accidentally or deliberately released in the greater Miami area. Several museum specimens of this toad collected in the Miami area 25 April 1979 had nymphal and adult A. rotundatum attached. Subsequent examination of living giant toads collected at another Miami area site from 1983 through 1985 revealed larval, nymphal, and adult A. rotundatum and confirmed colonization of this tick. Under laboratory conditions, another neotropical amphibian and reptile tick, Amblyomma dissimile, is capable of transmitting Cowdria ruminantium, the causative agent of heartwater, a disease present in the Caribbean area. Therefore, we suggest that A. rotundatum should also be tested for vectorial competence.

The Ornithodoros (Alectorobius) capensis group (Acarina: Ixodoidea: Argasidae) of the palearctic and oriental regions. O. (A.) maritimus: identity, marine bird hosts, virus infections, and distribution in western Europe and northwestern Africa.

In 1967, Vermeil and Marguet described Ornithodoros coniceps maritimus from larvae reared from larvae taken from marine birds on Dumet Island (Atlantic Ocean), Basse Bretagne, France. We collected O. (A.) coniceps Canestrini, 1890, from the type locality (Venice, Italy) and determined that the taxons coniceps and maritimus each require full species status. We selected a lectotype and paralectotypes for the taxon maritimus from the original Dumet Island material. The larva of maritimus is redescribed and the nymph, male, and female are described for the first time. Collection data are recorded from Dumet and other islands off France, Aegimures Islands off Tunisia, Puffin Island off northern Wales, and Great Saltee Island off Ireland. This tick infests nesting colonies of the common tern, roseate tern, sandwich tern, herring gull (northern and Mediterranean races), common cormorant, shag, razorbill, common murre, black-legged kittiwake, and probably other marine birds nesting nearby. Adults and nymphs (tentatively identified as maritimus but lacking associated larvae for full confirmation) were taken near nests of the little egret in Lake Tunis, Tunisia. Soldado virus was isolated from Puffin Island tick samples and a Soldado-like virus from Great Saltee Island tick samples. An experimental study of West Nile virus in the Tunisian tick population is reviewed. The birds species associated with maritimus in each collecting locality, and their nesting and resting habits and migration patterns in relation to tick and arbovirus survival and distribution, will be reported in the following paper in this series.

HARD TICK CALRETICULIN (CRT) GENE CODING REGIONS HAVE ONLY ONE INTRON WITH CONSERVED POSITIONS AND VARIABLE SIZES

Calreticulin (CRT ) is a unique eukaryotic gene. The CRT gene product, calreticulin, was first identified as a calcium binding protein in 1974, but further investigations have indicated that CRT protein performs many functions in cells, including involvement in evading the hosts immune system by parasites. Many studies of CRT have been published since the molecule was first discovered; however, the CRT gene exon-intron structure is only known for a limited number of ectoparasite species. In this study, we compared tick CRT genomic sequences to the corresponding cDNA from 28 species and found that 2 exons and 1 intron are present in the tick CRT gene. The intron position is conserved in 28 hard ticks, but intron size and nucleotide sequences vary. Three tick introns possess duplicated fragments and are twice as long as other introns. All tick CRT introns obey the GT-AG rule in the splice-site junctions and are phase 1 introns. By comparing tick CRT introns to those of fruit fly, mouse, and human, we conclude that tick CRT introns belong to the intron-late type. The number and size of CRT introns have increased through the evolution of eukaryotes.

CLONING AND SEQUENCING OF PUTATIVE CALRETICULIN COMPLEMENTARY DNAs FROM FOUR HARD TICK SPECIES

Calreticulin (CRT) is a calcium-binding protein and has many functions in eukaryotic cells. CRT is possibly involved in parasite host immune system evasion. To better understand the molecular basis of CRT in ticks, we cloned and sequenced 4 full-length complementary DNAs (cDNAs) from the hard tick species, Dermacentor variabilis, Haemaphysalis longicornis, Ixodes scapularis, and Rhipicephalus sanguineus, using the technique of rapid amplification of cDNA ends. The deduced amino acid sequences share high identities (between 77 and 98%) with 3 known tick CRT sequences. The major characteristics of known CRTs are observed in all 4 of our deduced tick CRTs. These include 3 major domains, a signal peptide sequence at the beginning of the coding region, 2 triplets of conserved regions, cysteine sites providing disulfide bridges for N-terminal folding, and a nuclear localization signal. Remarkably, the replacement of the endoplasmic reticulum retention signal KDEL by HEEL, which is believed to be associated with secretion of CRT into the host during feeding and was previously recorded only in 2 ticks and a hookworm, is also present in all 4 of our tick putative CRTs. In addition, the CRT gene is potentially useful for tick phylogenetic reconstruction.