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

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

Life-cycle and host specificity of Amblyomma tigrinum (Acari: Ixodidae) under laboratory conditions

Biological data of three generations of Amblyomma tigrinum in the laboratory are reported and the suitability of different host species for immature ticks are compared. Grouping the three generations, infestations by both the larval and nymphal stages were performed on chickens (Gallus gallus), wistar rats (Rattus norvegicus), rabbits (Oryctolagus cuniculus),wild mice (Calomys callosus), dogs (Canis familiaris) and opossums (Didelphis albiventris). Only dogs were used for infestations by adult ticks. Tick developmental periods were observed in an incubator at 27°C and RH 90%. The proportion of engorged larvae recovered from chickens (21.7% of the exposed larvae) was significantly larger (p<0.001) than those from the five mammal species used in the infestations (maximum of 3.1%). A significant larger (p<0.01) proportion of engorged larvae successfully molted after being fed on chickens than on mammal hosts. The proportion of engorged nymphs recovered from chickens (28.8% of the exposed nymphs) was significantly larger (p<0.001) than those from mammal hosts (range: 0–2.1%). Larvae showed similar feeding periods on exposure to different host species, except for those larvae fed on C. callosus, which showed significantly longer (p<0.001) feeding periods. Engorged larvae detachment peaked on the 5^th feeding day, followed by the 6^th day, on all hosts except for C. callosus. Larval premolt periods were similar for engorged ticks exposed to different host species, except for larvae fed on dogs, which showed significantly longer (p<0.001) premolt periods. Host detachment of engorged nymphs peaked on the 6^th feeding day on chickens. Although nymphal detachment on rats peaked on the 8^th day, only 15 nymphs were recovered from this host species. In a sample of 144 F₃ nymphs fed on chickens no significant difference (p>0.10) was found between the feeding or premolt periods of 82 males and 62 females, but female nymphs were significantly heavier (p<0.005) than male nymphs. Sixteen engorged females (61.5% of the exposed ticks) were recovered after being fed on dogs, and all these females laid viable eggs. Chickens, the only avian host, were the most suitable host when compared with the five mammal species. Dogs were demonstrated to be a suitable host for adults of A. tigrinum, which is consistent with, several reports of adult A. tigrinum ticks parasitizing dogs in different areas of South America. Our results reinforce that in these same areas avian species are the major hosts for immature stages of this tick species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Prevalence of tick-borne pathogens in ixodid ticks (Acari: Ixodidae) collected from European wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>) and Iberian red deer (<Emphasis Type="Italic">Cervus elaphus hispanicus</Emphasis>) in central Spain

Emerging tick-borne diseases of humans and animals have occurred frequently during the past 30 years. These disease outbreaks appear to result from changes in the distribution of tick and vertebrate hosts, and the introduction of humans and domestic animals into tick–pathogen–wildlife cycles. Use of molecular technologies now available for identification of pathogens in ticks can provide valuable information that allows for risk analysis of emerging tick-borne diseases. In this study, the prevalence of selected pathogens in ticks collected in six locations in central Spain from the major wild ungulate species, European wild boar (Sus scrofa) and Iberian red deer (Cervus elaphus hispanicus), was determined by PCR. Tick species collected included Ixodes ricinus, Dermacentor marginatus, Rhipicephalus bursa and Hyalomma m. marginatum. Pathogens identified in ticks included piroplasmids, Anaplasma spp., Ehrlichia spp. and Rickettsia spp. Piroplasmids were identified in all tick species except I. ricinus. Ehrlichia spp. were detected in all tick species and collection locations, while Rickettsia spp., which proved to be R. slovaca and a recently identified Rickettsia sp. DnS28, were identified only in D. marginatus. A. marginale and A. phagocytophilum were detected in D. marginatus, R. bursa and Hy. m. marginatum. Concurrent infections of these pathogens were frequently observed in ticks. Notably, A. phagocytophilum, which is infective for a broad host range that includes humans and domestic and wild animals, was identified in ticks from all collection locations. The variety of ticks and tick-borne pathogens demonstrated in this study suggests a risk in central Spain for the emergence of tick-borne diseases in humans and domestic animals.     

Amphibian survival,growth and development in response to mineral nitrogen exposure and predator cues in the field: an experimental approach

Mineral nitrogen (N) has been suggested as a potential factor causing declines in amphibian populations, especially in agricultural landscapes; however, there is a question as to whether it remains in the water column long enough to be toxic. We explored the hypothesis that mineral N can cause both lethal and sublethal toxic effects in amphibian embryos and larvae in a manipulative field experiment. We sampled 12 ponds, fertilizing half with ammonium nitrate fertilizer early in the spring, and measured hatching, survival, development, growth, and the incidence of deformities in native populations of wood frog (Rana sylvatica) and eastern tiger salamander (Ambystoma tigrinum tigrinum) embryos and larvae held in in situ enclosures. We found that higher ammonium concentrations negatively affect R. sylvatica more strongly than A. tigrinum. R. sylvatica tended to have lower survival as embryos and young tadpoles, slowed embryonic development, and an increased proportion of hatchlings with deformities at experimentally elevated ammonium. A. tigrinum did not experience significantly reduced survival, but their larval development was slowed in response to elevated ammonium and the abundance of large invertebrate predators. Variable species susceptibility, such as that shown by R sylvatica and A. tigrinum, could have large indirect effects on aquatic community structure through modification of competitive or predator-prey relationships. Ammonium and nitrate + nitrite concentrations were not correlated with other measures that might have affected amphibians, such as pH, pond area, depth, or vegetation. Our results highlight the potential importance of elevated ammonium on the growth, development and survival of amphibians, especially those that breed in surface waters receiving anthropogenic N inputs. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The natural hosts for larvae and nymphs of Amblyomma neumanni and Amblyomma parvum (Acari: Ixodidae)

Based on the hypothesis that birds and rodents are important hosts for subadults of the Neotropical Amblyomma neumanni and Amblyomma parvum ticks, a survey of these type of hosts was carried out from July 2004 to March 2006, in Quilino (A. parvum) and Dean Funes (A. neumanni), Córdoba province, Argentina. Additionally, monthly tick counts were performed on cattle and goats with occasional tick search in other domestic hosts. Records of questing height of subadult ticks on vegetation were also carried out monthly. Rodents (n = 123) and birds (n = 122) captured in Dean Funes showed no infestation with A. neumanni. Apart of few nymphs found on horses, all larvae and nymphs of A. neumanni were on cattle with a larval prevalence and mean number of 22.2%, and 7.7 ± 22.52, respectively, and a prevalence of nymphs of 47.8% with a mean of 7.9 ± 18.49. The average questing height of larvae and nymphs of A. neumanni was 23.5 ± 17.1 cm and 30.7 ± 26.7 cm, respectively. A total of 138 rodents and 130 birds were captured in Quilino but the Caviidae rodent Galea musteloides carried 99.3% of larvae and 99.8% of nymphs of A. parvum, and no immature stages were detected on cattle, goat or vegetation. Tick counts on G. musteloides (n = 74) showed a prevalence of 42% and a mean number of 9.9 ± 24.83 for larvae, while nymphal infestation had a prevalence of 56.5% and a mean of 8.7 ± 11.31. Cattle appear to be suitable hosts to sustain the complete cycle of A. neumanni in nature (adult ticks infest cattle too) and questing height of subadults indicates that they are expecting to feed on medium and large-sized mammals, such as cattle and other ungulates. At least in the study site, G. musteloides is the principal host for the survival strategy of A. parvum subadults; adult ticks are common on cattle and goats. These hosts are introduced in the Neotropics but A. neumanni was able to develop a surrogate cycle independent of native hosts while A. parvum still depends on probably primeval hosts to sustain their larvae and nymphs.     

Hosts and distribution of Amblyomma auricularium and Amblyomma pseudoconcolor Aragão, 1908 (Acari: Ixodidae)

Collections of Amblyomma auricularium and A. pseudoconcolor Aragão, 1908 are discussed in relation to distribution and hosts. Three tick collections (two from Argentina and a third from the USA) house a total of 574 A. auricularium (307 males, 162 females, 73 nymphs and 32 larvae) and 179 A. pseudoconcolor (96 males, 74 females, 4 nymphs and 5 larvae). Apart from an adult A. pseudoconcolor found on a bird, Nothura maculosa Temminck, 1815, all ticks were found on mammals. The great majority of specimens of both ticks species were removed from the family Dasypodidae Gray, 1821 (84.9% and 93.8% of A. auricularium and A. pseudoconcolor, respectively). Amblyomma auricularium has also been found on wild hosts of the families Myrmecophagidae and occasionally Didelphidae, Caviidae, Chinchillidae, Hydrochaeridae, Muridae, Canidae, Mustelidae, Procyonidae and domestic animals (cattle, dogs, horses), while A. pseudoconcolor has also been found occasionally on wild hosts of the family Didelphidae and on domestic animals (cattle, dogs). Amblyomma pseudoconcolor appears to be restricted to the Neotropical region, covering northern Argentina and the eastern region of South America from Uruguay to Surinam, including south-eastern Paraguay, eastern Brazil and French Guiana. Amblyomma auricularium is distributed from northern Patagonia in Argentina throughout the Neotropics into the Nearctic region up to the southern USA (Texas, Florida), with collection localities also in Belize, Bolivia, Brazil, Colombia, Costa Rica, French Guiana, Honduras, Mexico, Nicaragua, Panama, Paraguay, Trinidad & Tobago, Uruguay and Venezuela. It is not known whether A. auricularium is an established resident of the USA.     

Cattle ticks of the genera <Emphasis Type="Italic">Rhipicephalus</Emphasis> and <Emphasis Type="Italic">Amblyomma</Emphasis> of economic importance in Tanzania: distribution assessed with GIS based on an extensive field survey

In order to implement a robust integrated tick and tick-borne disease control programme in Tanzania, based on ecological and epidemiological knowledge of ticks and their associated diseases, a national tick and sero-surveillance study was carried out in all 21 regions of the mainland, as well as on Mafia Island, between 1998 and 2001. The current distributions of Rhipicephalus appendiculatus, R. pravus, Amblyomma variegatum, A. gemma, and A. lepidum are illustrated and discussed. Tick distribution maps were assessed using the Weights-of-Evidence method (WofE), and employing temperature, humidity, NDVI, rainfall, and land-cover predictive data. Ground-truthing was done to check correspondence both of the data employed in prediction with land-cover characteristics discerned in the field as well as of the surveyed and predicted tick distributions. Statistical methods were used to analyse associations of the tick species with their environment, cattle density, and other ticks. Except for R. appendiculatus, no appreciable changes were demonstrated in the predicted and observed tick distributions compared to the existing maps that originated in the 1950–1960s. Cattle density influenced the distribution of A. variegatum and, to a certain extent, of A. lepidum, but had no appreciable influence on the distribution of any of the other ticks discussed in this paper, neither did livestock movement. Distinct differences for environmental requirements where observed between different tick species within the same genus. The predictive maps of R. appendiculatus and R. pravus suggest their mutually exclusive distribution in Tanzania, and simultaneous statistical analysis showed R. pravus as a greater specialist. Of the three Amblyomma species, A. variegatum is the most catholic tick species in Tanzania, while both A. gemma and A. lepidum belong to the more specialized species. Despite dissimilar habitat preferences, all three Amblyomma spp. co-exist in central Tanzania, where very heterogeneous habitats may simultaneously satisfy the environmental requirements of all three species. The current study, conducted about 4 decades after the last major survey activities, has shown that changing livestock policies, unrestricted livestock movement and a continuous change in climatic/environmental conditions in Tanzania have brought about only limited changes in the distribution patterns of R. appendiculatus, R. pravus and the three Amblyomma species investigated. Whether this observation indicates a relative indifference of these ticks to environmental and/or climate changes allows room for speculation.     

The relationships between Ixodes ricinus and small mammal species at the woodland–pasture interface

Ixodes ricinus, as vector, and small mammals, as reservoirs, are implicated in pathogen transmission between wild fauna, domestic animals and humans at the woodland–pasture interface. The ecological relationship between ticks and small mammals was monitored in 2005 on four bocage (enclosed pastureland) sites in central France, where questing ticks were collected by dragging and small mammals were trapped. Questing I. ricinus tick and small mammal locations in the environment were assessed through correspondence analysis. I. ricinus larval burden on small mammals was modeled using a negative binomial law. The correspondence analyses underlined three landscape features: grassland, hedgerow, and woodland. Seven small mammal species were trapped, while questing ticks were all I. ricinus, with the highest abundance in woodland and the lowest in pasture. The small mammals were overall more abundant in hedgerow, less present in woodland and sparse in grassland. They carried mainly I. ricinus, and secondarily I. acuminatus and I. trianguliceps. The most likely profile for a tick-infested small mammal corresponded to a male wood mouse (Apodemus sylvaticus) in woodland or hedgerow during a dry day. A. sylvaticus, which was the only species captured in grassland, but was also present in hedgerow and woodland, may be a primary means of transfer of I. ricinus larvae from woodland to pasture.     

The seasonal occurrence of ticks (Acari: Ixodidae) on sheep and in the field in the Judean area of Israel

A 2 year survey of ixodid ticks in the Judean area of Israel between 1983 and 1985 showed that sheep were parasitized by the following species:Rhipicephalus sanguineus group (30.6%),Rhipicephalus bursa (25.3%),Haemaphysalis cretica (20.5%),Haemaphysalis otophila (20.1%) andHyalomma anatolicum excavatum (3.1%). Three other species,Hyalomma marginatum rufipes, Boophilus annulatus andAmblyomma lepidum were also present but in very low numbers. A total of 50 816 specimens (78.1% adult ticks) were collected from the sheep. In the field, a total of 4780 ticks (8.7% adults) were collected and 64.3% consisted ofR. bursa. Other species found wereH. cretica (14.5%),H. a. excavatum (13.7%),H. otophila (3.4%),R. sanguineus group (1.9%) and 1.7% wasIxodes eldaricus. The number of ticks collected varied from year to year and seasonal activity was observed in all species except forH. a. excavatum. In all species, males outnumbered females, with the exception ofH. cretica. The seasonal occurrence, tick ecology and disease relationship is discussed for each species.     

Acquisition of Borrelia burgdorferi by Ixodes ricinus ticks fed on the European hedgehog,Erinaceus europaeus L

A hedgehog, Erinaceus europaeus, was found to be heavily infested with larval and nymphal Ixodes ricinus in a forest park in Co. Galway, Ireland. A large proportion of the ticks that engorged and detached were infected with the spirochacte, Borrelia burgdorferi, the causative agent of human Lyme borreliosis. The identity of these spirochaetes was confirmed by immunofluorescent assay with B. burgdorferi-specific monoclonal antibody and by polymerase chain reaction test and they were transmitted from the hedgehog to laboratory-reared ticks and from the ticks obtained from the hedgehog to gerbils (Meriones unguiculatus). The high infection rate of the larvae that fed on the hedgehog in comparison with unfed larvae from the same habitat was interpreted as strong evidence that this host species is reservoir competent. Since hedgehogs can evidently feed adult ticks as well as many immature stages, they may well have an important role in the ecology of Lyme borreliosis in some habitats.     

Natural attachment duration of nymphs of the tick Ixodes uriae (Acari: Ixodidae) on kittiwake Rissa tridactyla nestlings

Daily searches of the plumage of 62 kittiwake (Rissa tridactyla) chicks were carried out for 3 weeks to determine the attachment duration of the tick Ixodes uriae. All ticks recorded were nymphs and their mean duration of attachment was 5.2±1.7 days (n=93). Seventy-four per cent of the ticks survived to engorgement and were assumed to have fed successfully. The mean duration of attachment of successful ticks was 5.8±1.0 days (n=69), significantly longer than that of unsuccessful ticks (3.3±2.1 days, n=24). There was no difference in survival rates to engorgement between ticks attached to feathered and unfeathered parts of the body (78.3 and 79.2%, respectively), but the duration of attachment of successful ticks was significantly longer on unfeathered compared to feathered areas. There were no significant differences in the survival to engorgement or duration of attachment between ticks found on young (c.11 days) and old (>11 days) kittiwake nestlings. The attachment duration was not related to the total number of ticks found on the host or the number of ticks present in the immediate vicinity of the site of attachment. This work provides important data on the parameter of attachment durations of nymphal I. uriae on free-living kittiwakes; the data can be used for incorporation into the application of population modelling.     

How Specific are Host-Produced Kairomones to Host-Seeking Ixodid Ticks?

Ixodid ticks respond to host-produced substances (kairomones) that influence the ticks’ host-finding behavior. In the laboratory adult blacklegged ticks, Ixodes scapularis Say, lone star ticks, Amblyomma americanum L., and American dog ticks, Dermacentor variabilis (Say) became akinetic on residues rubbed from their principal hosts (deer for the former two species and dogs for the latter). However, arrestment also occurred when adults of these species were tested using the same method bioassay, but with host substances reversed (i.e., I. scapularis and A. americanum against canine substances, and D. variabilis against deer gland substances). Although adult D. variabilis exhibited arrestant responses to deer substances and are often found along trails used by deer, they apparently make little use of deer as hosts. It is unclear whether responding to deer-produced kairomones may have disadvantages for D. variabilis. Until the active components of host-produced arrestment kairomones are isolated, identified and evaluated in behavioral tests, this host-finding strategy remains only partially understood. This revised version was published online in July 2006 with corrections to the Cover Date.     

Experimental infection of the rabbit tick,<Emphasis Type="Italic"> Haemaphysalis leporispalustris</Emphasis>, with the bacterium<Emphasis Type="Italic"> Rickettsia rickettsii</Emphasis>, and comparative biology of infected and uninfected tick lineages

The present study consisted of two experiments that evaluated experimental infections of Haemaphysalis leporispalustris ticks by a Brazilian strain of Rickettsia rickettsii, and their effect on tick biology. In experiment I, ticks were exposed to R. rickettsii during the larval, nymphal or adult stages by feeding on rabbits (Oryctolagus cuniculus) needle-inoculated with R. rickettsii, and thereafter reared on uninfected rabbits for the entire next tick generation. Regardless of the tick stage that acquired the infection, all subsequent tick stages were shown to be infected by PCR (infection rates varying from 1.3 to 41.7%), and were able to transmit R. rickettsii to uninfected rabbits, as demonstrated by rabbit seroconversion, guinea pig inoculation with rabbit blood, and PCR on rabbit blood. In Experiment II, ticks were exposed to R. rickettsii during the larval stage by feeding on rabbits co-infested with R. rickettsii-infected adult ticks, and thereafter reared on uninfected rabbits until the next generation of larvae. Again, all subsequent tick stages were shown to be infected by PCR (infection rates varying from 3.0 to 40.0%), and were able to transmit R. rickettsii to uninfected rabbits. Thus, it was demonstrated that larvae, nymphs, and adults of H. leporispalustris were able to acquire and maintain the R. rickettsii infection by transstadial and transovarial transmissions within the tick population, with active transmission of the bacterium to susceptible rabbits by all parasitic stages. Analyses of biological parameters of uninfected and R. rickettsii-infected tick lineages were performed in order to evaluate possible deleterious effects of R. rickettsii to the infected tick lineages. Surprisingly, all but one of the four R. rickettsii-experimental groups of the present study showed overall better biological performance than their sibling uninfected control ticks. Results of the present study showed that H. leporispalustris could support infection by a high virulent strain of R. rickettsii for at least two generations, in which infected tick lineages tended to have better performance than uninfected ticks. Our results support a possible role of H. leporispalustris in the enzootic maintenance of R. rickettsii in Latin America, as previously suggested by earlier works.     

Preliminary studies on the effect of Anaplasma marginale antibodies ingested by Dermacentor andersoni ticks (Acari:Ixodidae) with their blood meal on infections in salivary glands

The effect of Anaplasma marginale antibodies ingested with the tick blood meal was tested on infected male ticks that were allowed to feed on cattle immunized with the erythrocytic stage of A. marginale. The experiments were done in two trials. Trial 1 was done using splenectomized calves (two calves per treated and control groups) while ticks in trial 2 were fed on intact yearling cattle (four cattle per treated and control groups). The cattle were immunized with purified outer membrane proteins of erythrocyte-derived A. marginale using saponin (trial 1) or monophosphoryl lipid-A-trehalose dicorynomycolate adjuvant (trial 2). The corresponding control cattle received adjuvant only. All cattle were challenged using Dermacentor andersoni males infected as adults that were allowed to feed for 7 days. In trial 1, the ticks were allowed to feed a second time on susceptible calves to test whether exposure of ticks to immunized cattle affected their ability to transmit anaplasmosis. Infections in fed ticks were monitored by determining the infection rates in salivary glands with an A. marginale-specific RNA probe and light microscopy. Vaccine-derived antibodies ingested with the tick blood meal did not appear to affect the development of A. marginale in previously infected ticks. The infection rates in the salivary glands were not significantly different among ticks fed on immunized versus adjuvant control cattle. When the vaccine-exposed ticks in trial 1 were allowed to feed a second time on susceptible calves, the resulting clinical symptoms of anaplasmosis were similar to those of the controls. There was no statistically significant effect of tick exposure to the anti-erythrocytic stage antibody on the development of salivary gland infection or transmission of A. marginale by ticks.     

Ticks (Acari: Ixodidae) within various phytophysiognomies of a Cerrado reserve in Uberlandia, Minas Gerais, Brazil

Cerrado biome, the South American savannah, covers about 2 million km² and is very rich in endemic species but threatened by agriculture. In this report free-living tick species are presented, and their seasonal and relative distribution within the various phytophysiognomies in a small Cerrado reserve in Minas Gerais State, Brazil. Overall 2,694 free-living ticks were found during a 2 years sampling period with CO₂ traps and cloth dragging. Of these, 73.5% were Amblyomma cajennense and 0.6% Amblyomma dubitatum. All other ticks (25.9%) were retained as Amblyomma spp. Adults of A. cajennense peaked in spring, the nymphs in winter of both years. Amblyomma larval clusters were found in autumn and winter. Adult ticks (46.7%) and nymphs (39.5%) were most often found in woodlands, whereas most larval clusters were found in valley-side marshes (39%). Amblyomma cajennense, Anocentor nitens, Rhipicephalus (Boophilus) microplusand Rhipicephalus sanguineus ticks were found on domestic animals from neighboring properties. Search for Rickettsia in the hemolymph of 497 A. cajennense and one A. dubitatum ticks yielded negative results. Results confirmed earlier reports on the overwhelming prevalence of A. cajennense ticks in the Cerrado biome of Brazil and added information to habitat preferences of this tick species, a major vector in Brazil of the Rocky Mountain spotted fever.     

Adaptations of the Tick-Borne Pathogen, Anaplasma marginale, for Survival in Cattle and Ticks

The tick-borne cattle pathogen Anaplasma marginale (Rickettsiales: Anaplasmataceae) multiplies within membrane-bound inclusions in host cell cytoplasm. Many geographic isolates of A. marginale occur that vary in genotype, antigenic composition, morphology and infectivity for ticks. A tick cell culture system for propagation of A. marginale proved to be a good model for study of tick-pathogen interactions. Six major surface proteins (MSPs) identified on A. marginale from bovine erythrocytes were conserved on A. marginale derived from tick cells. MSP1a and MSP1b were adhesins for bovine erythrocytes, while only MSP1a was found to be an adhesin for tick cells. The tandemly repeated portion of MSP1a was found to be necessary and sufficient for adhesion to both tick cells and bovine erythrocytes. Infectivity of A. marginale isolates for ticks was dependent on the adhesive capacity of the isolate MSP1a, which was found to involve both the adhesive properties and sequence of the repeated peptides. Cattle immunized with A. marginale derived from bovine erythrocytes or tick cells demonstrated a differential antibody response to MSP1a and MSP1b that resulted from the differential expression of these proteins in cattle and ticks cells. MSP2, derived from a multi-gene family, was found to undergo antigenic variation in cattle and ticks and may contribute to establishment of persistent A. marginale infections. MSP1a has been used as a stable genetic marker for geographic isolates because the molecular weight varies due to differing numbers of the tandem repeats. However, phylogenetic studies of A. marginale isolates from North America using MSP1a and MSP4 demonstrated that MSP4 was a good biogeographic marker, while MSP1a varied greatly among and within geographic areas. Infection and development of A. marginale in cattle and tick cells appears to differ and to be mediated by several surface proteins encoded from the small genome. This revised version was published online in July 2006 with corrections to the Cover Date.     

Rhipicephalus appendiculatus: characterization of a testis-associated protein

A novel gene coding for Rhipicephalus appendiculatus Male-specific Protein (RAMP) was identified in a cDNA library constructed from the testis/vas deferens of R. appendiculatus ticks. This gene encodes a secreted protein exclusively expressed in the testis/vas deferens. The putative RAMP amino acid sequence contains a signal peptide and has 29% amino acid identity with male-specific Is5 gene of Ixodes scapularis. Gene expression studies revealed that RAMP mRNA was up-regulated in male ticks during blood feeding. RAMP was detected not only in the testis/vas deferens of males but also in postcoitum female ticks based on Western blotting, indicating that this protein is transferred to the female tick during copulation. Virgin female ticks, microinjected with recombinant RAMP, had significantly prolonged attachment duration during feeding, but there was no effect on fed weight. These results suggest that RAMP is a male-specific molecule in the spermatophore, and is related to female attachment behavior in R. appendiculatus ticks.     

Drop off rhythm and survival periods of <Emphasis Type="Italic">Amblyomma lepidum</Emphasis> (Acari: Ixodidae) under field conditions

In this study, engorged Amblyomma lepidum ticks were found to drop off in two peaks, one in the morning and one in the evening. Most larvae and females engorged during the morning hours between 06.00 h and 10.00 h with a peak around 08.00 h, whereas the majority of the nymphs dropped in the evening between 18.00 h and 24.00 h with the peak around 22.00 h. Although the effect of time on drop-off patterns of the ticks was statistically significant (p≤ 0.001), there were no significant seasonal influences. Survival of unfed stages of A. lepidum was also studied and was found to increase from larvae to adult ticks. The longest survival periods of 10, 11 and 14 weeks were recorded during the wet season for larvae, nymphs and adults, respectively. It is concluded that environmental conditions required for survival of A. lepidum are optimal only during the wet season and that during other seasons the tick depends primarily on prevailing micro-climatic conditions for its survival.     

Immunity against Boophilus annulatus induced by the Bm86 (Tick-GARD) vaccine

Friesian cattle were immunized with two inoculations of anti-tick Bm86 (Tick-GARD) vaccine and were challenged 30 or 90 d later with Boophilus annulatus larvae derived from 1.2 g of eggs. No nymphs or adult ticks were found on the immunized cattle during four weeks after challenge. Repeated infestations (2 to 4) with larvae on three other calves during a period of 160 and 390 d after the immunization did not result in development of nymphal and adult stages. In control, non-immunized cattle infested with corresponding batches of larvae 1380 to 4653 replete adult female ticks were collected. Larvae issued from Babesia bovis-infected female ticks transmitted the infection to Bm86-immunized cattle, but the progeny of B. bigemina-infected females did not. Since B. bigemina is transmitted exclusively by nymphal stages of Bo. annulatus these results support the observation that immunity induced by Bm86 affects the larval stage of this tick.     

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

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