Variegatum Nymphs Research Articles

Zoonotic Pathogens Detected in Ticks in Kenyan Game Reserves

Little is known on tick-borne pathogens and their role in disease in game reserves in Kenya. Ticks were collected by sterile forceps from restrained cattle hide and placed into labeled falcon tubes. Ticks were screened for pathogens by High Resolution Melting (HRM) analysis and sequencing of specific RT-PCR products of Anaplasma, Ehrlichia, and Rickettsia species. A total of 317 ticks (281 adult ticks and 36 nymphs) comprising seven species were collected around the Tsavo National Reserve (TNR) in Taita Taveta County with Amblyomma gemma being the most commonly collected species (n = 135, 42.6%). From near Shimba Hill game reserve (SHNR), a total of 240 adult’s ticks were sampled, representing eight species, with again Amblyomma gemma being the most sampled species (n = 156, 65%). From Tsavo, a total of three pools of Rhipicephalus appendiculatus were positive for Theileria parva, two pools of Rhipicephaline evertsi for Anaplasma platys and one pool of Amblyomma variegatum nymphs for Rickettsia africae. Rickettsia africae, which causes African tick-bite fever, was detected in two pools of Am. variegatum and one pool of Amblyomma gemma collected near Shimba Hill game reserve. Rickettsia sp. and Anaplasma sp. were detected in Am. gemma and Rh. evertsi respectively. Rickettsia aeschlimannii was detected in a pool of Am. gemma. These findings highlight the risk of transmission of zoonotic pathogens to humans in regions with high human-wildlife interfaces. Of specific importance, we provide evidence of R. aeschlimannii in A. gemma for the first time, representing a potential new R. aeschlimannii vectors.

The potential of Rhipicephalus microplus as a vector of Ehrlichia ruminantium in West Africa

Heartwater, or cowdriosis, is a virulent tick-borne rickettsial disease of ruminants caused by Ehrlichia ruminantium, biologically transmitted by Amblyomma species (A. variegatum in West Africa). In West Africa, this bacterium was recently reported to naturally infect the invasive cattle tick, Rhipicephalus microplus (Rm) through trans-ovarian transmission from replete adult females to offspring. A ‘sheep-tick-sheep’ cycle was set up to determine whether feeding the progeny of these ticks on naïve sheep could lead to infection, and to compare clinical outcomes resulting from this transmission with those observed following infection by the natural A. variegatum (Av) vector. Using local strains of ticks (KIMINI-Rm and KIMINI-Av) and of E. ruminantium (BK242), we recorded, using the PCR technique, the presence of bacterial DNA in ticks (larvae for Av and females for Rm) engorged on sheep inoculated by BK242-infected blood. The bacterial DNA was also detected in the next stages of the lifecycle of R. microplus (eggs and larvae), and in sheep infested either by those R. microplus larvae or by A. variegatum nymphs moulted from larvae engorged on blood-inoculated sheep. Bacterial infection in these sheep was demonstrated by detecting antibodies to E. ruminantium using the MAP1-B ELISA and by isolation of the bacterium on cell culture from blood. The sequences of PCS20 gene detected in ticks and sheep were identical to that of the BK242 strain. Our results confirm that R. microplus can acquire and transmit E. ruminantium to the next stage. However, this transmission resulted in a mild subclinical disease whereas severe clinical disease was observed in sheep infested by A. variegatum infected nymphs, suggesting differences in the tick/bacteria relationship. Future studies will focus on replicating these findings with ticks of different isolates and life stages to determine if R. microplus is playing a role in the epidemiology of heartwater in West Africa. Additionally, studies will investigate whether sheep that are seropositive due to infestation by E. ruminantium-infected R. microplus are subsequently protected against heartwater. Such data will add to our understanding of the possible impact of R. microplus in areas where it has become recently established.

Prevalence of ectoparasites attacking helmet guineafowl, <i>Numidamele agridis</i>, in Sokoto, north-western Nigeria

A study was conducted to determine the prevalence of ectoparasites infesting the grey-breasted guinea fowls in Sokoto, north-western Nigeria for a period of one year, July 2010 to June 2011. In the study, the ectoparasites were recovered from the birds by dust-ruffling technique and identified using light microscope following clearing in lactophenol. From a total of 165 birds of various ages examined, 19 species of ectoparasites were recovered. They included 9 species of chewing lice as follows: Menacanthus cornutus with a prevalence of 84%, Lipeurus tropicalis 68%, Lipeurus caponis 20%, Amyrsidea powelli 80%, Goniodes gigas 70%, Goniodes fimbriatus 70%, Goniocotes gallinae 68%, Cuclotogas teroccidentalis 10%, Numidicolaan tennatus 45%;six species of mites; Rivoltasiabi furcata 20%, Epidermoptes bilobatus 20%, Megniniacu bitalis/Megninia tetraonis 65%, Bdellonys susbursa 45%, Cnemidocop tesmutans 15%; three species of ticks: Argaspersicus (Iarvae) 30%, Amblyomma variegatum (larvae and nymphs) 6%, Haemphy salishoodi 4%, and a species of flea: Echidno phagagallinacea 12%. Ectoparatises fluctuated seasonally with overall prevalence higher in the hot dry season ( p <0.01) and warm wet season ( p <0.05) than the cool dry harmattan season. It was striking that classical clinical infestations were seldom seen despite widespread occurrence of heavy burdens. Keywords: ectoparasites, guineafowls, semi-arid area, Sokoto, seasons

Survival and moulting of Amblyomma variegatum nymphs under cold conditions of the Malagasy highlands

Although Amblyomma variegatum is now regularly recorded up to 1600m in altitude in the Malagasy highlands, where it was previously reported not to persist without a constant supply of ticks introduced from lower infested regions, some parts of the highlands remain tick-free. Studies were carried out to verify whether the cold climate prevailing in these areas in June–September could prevent the survival and moulting of nymphs, the tick life stage present in the environment at this period. Cohorts of engorged A. variegatum nymphs were released from June to August in six different sites (three in 2010, altitudes 1200–1415m; three in 2011, altitudes 1585–1960m) which were reported to be either tick-infested (two in 2010, one in 2011) or tick-free. The ticks were placed in cages driven into the soil and open at the bottom so that they could hide in the soil or root network. Of the 1975 nymphs released in 2010 and the 1494 released in 2011, 86% and 85% were recovered, respectively. Twenty to 23% of the recovered ticks were dead, and some of them were obviously predated; predation also likely contributed to the disappearance of the non-recovered ticks. When the rainy season started in October, 59% of the newly moulted adults were still alive in the cages. The moulting period lasted up to 20weeks, depending on the site and release period. As verified in 2011, unfed nymphs could also survive the cold season. Various A. variegatum life stages are thus able to survive the adverse cold and/or dry seasons: unfed nymphs, engorged nymphs in developmental diapause, moulted adults in behavioural diapause as observed previously. Strong variation in mortality and recovery rates was observed between cages, highlighting the importance of the micro-environment and micro-climate for tick survival. The minimum temperature recorded in the field sites varied from 1.1°C to 6.8°C, but the tick-free sites were not the coldest ones; they were, however, those for which the temperature remained below 10°C for the longest time over the study period. Recovery and mortality rates in the tick-free sites were similar to those of the tick-infested sites: the temperatures recorded during the study periods did not prevent ticks from surviving and moulting although it did delay the metamorphosis. Low temperature alone can therefore not explain the persistence of tick-free areas in the highlands. To further monitor survival, cohorts of engorged nymphs were also maintained in an incubator at 3.6°C, 6.2°C or 12.8°C. More than 50% mortality was observed after 6days at 3.6°C, and after 15days at 6.2°C, whereas 18days at 12.8°C only delayed moulting. The collected survival, moulting and climatic data presented in this study should help to develop a predictive model to assess the distribution of A. variegatum according to climate characteristics.

Moulting and survival of Amblyomma variegatum (Acari: Ixodidae) nymphs in quasi-natural conditions in Burkina Faso; tick predators as an important limiting factor

The survival rate of Amblyomma variegatum engorged nymphs was studied in 9 cohorts released during the first part of the dry season (i.e. the nymph infestation period in the region) and placed on a natural savannah, either in plastic containers or in cages driven into the soil and open at the bottom. All nymphs put in the containers placed in the sun died rapidly, while a high moulting rate (94%) was observed in containers placed in the shade. This rate reached 63% for the ticks released in the cages. The moulting period lasted from less than 4 to more than 6 weeks, depending on the release period: unfed adults are therefore present in the pasture 4-5 months before the onset of the adult infestation period. The survival rate of the unfed adults did not vary according to the release period of the nymphs: at the beginning of the rainy season, 20-25% of the adult ticks were still alive in the cages. Very important variations in survival and disappearance rates were observed according to the place where the cages had been installed: within a few metres, the survival rate could vary from 0 to 90%, presumably because of the activity of predators which removed also dead tick from the plastic containers. Predators may then have a great but highly variable and unpredictable impact on the natural population of A. variegatum adults. Engorged nymphs released on pasture were able to find suitable micro-habitats allowing survival, covered up to 2.8 metres to reach them, and grouped together at the neck or root network of bushes and grass clumps, sometimes burrowing to a depth of 10 cm.

Exploiting the heterogeneous drop-off rhythm of Amblyomma variegatum nymphs to reduce pasture infestation by adult ticks

Tick distribution depends on the drop-off rhythm of the previous stage and on the suitability for tick survival of the environment where they are disseminated. Studies were implemented in Burkina Faso to assess detachment pattern of engorged Amblyomma variegatum nymphs. Experiments were carried out with naturally infested cattle kept in a paddock or monitored when grazing community pasture. In the pasture, 80% of the nymphs detached between 14.00 h and 17.00 h while less than 25% did so in the paddock. Further investigation was implemented to assess whether the density of adult ticks might be modified by herd management. During the early dry season, zebus grazed in 4 plots fenced in natural savannahs and fallows. Two of the plots were used in the morning and the two others in the afternoon. Six months later, zebus were put in these plots, in turn, on 9 occasions. The number of A. variegatum adults picked up by the cattle in each plot was highly variable: they captured more ticks in the plots installed on good lush savannah and 3-fold more ticks in those where the herd had grazed in the afternoon during the previous dry season. An integrated tick control strategy taking these results into account is proposed.

Immunomodulatory arsenal of nymphal ticks

Ticks have developed their own immunomodulatory mechanisms to inhibit the host inflammatory response. One of them involves the ability to subvert the cytokine network at the site of tick feeding by secreting cytokine binding molecules. Most studies have focused on the immunomodulatory prowess of adult female ticks. Here we describe anti-cytokine activity in salivary gland extracts (SGEs) prepared from 2-day-fed nymphs of Dermacentor reticulatus Fabricius, Ixodes ricinus L., Rhipicephalus appendiculatus Neumann and Amblyomma variegatum Fabricius. Anti-CXCL8 activity was detected in nymphs of all species. Relatively high activity against CCL2, CCL3 and CCL11 was observed in SGEs of R. appendiculatus and A. variegatum nymphs, whereas SGEs of I. ricinus nymphs showed comparatively high anti-interleukin-2 (-IL-2) and anti-IL-4 activities. These data show that nymphs, which epidemiologically are usually more important than adults as disease vectors, possess a range of anti-cytokine activities that may facilitate pathogen transmission.

Life history of the tick parasitoid Ixodiphagus hookeri (Hymenoptera: Encyrtidae) in Kenya

We conducted laboratory and field experiments to elucidate the life history of Ixodiphagus hookeri, a parasitoid of the ixodid tick Amblyomma variegatum in Western Kenya. Ixodiphagus hookeri females oviposited in unfed host nymphs as well as engorged nymphs, but rarely in engorged larvae. While I. hookeri developed to adults in engorged nymphs, the eggs laid in unfed nymphs disappeared within 2 days after oviposition. As temperature increased, development time of I. hookeri from oviposition to adult emergence in engorged nymphs decreased from 46 days at 23 °C to 35 days at 28 °C, and their immature survival in engorged nymphs decreased from 67% at 23 °C to 22% at 28 °C. No parasitoid adult emerged from hosts at 30 °C. Individual hosts parasitized by single females produced 42–53 adult wasps, 73% of which were females. As a typical pro-ovigenic species, I. hookeri females had an average of 84 mature eggs at emergence and lived only for a few days. When laboratory-reared, unfed nymphs of A. variegatum were attached to cattle for 4–9 days in subsistence farmers’ fields in Western Kenya, 25% of the engorged nymphs and 4% of the unfed nymphs on cattle were parasitized by I. hookeri, demonstrating that I. hookeri females search for and oviposit in A. variegatum nymphs on cattle. Unlike other strains of I. hookeri that overwinter as eggs in unfed nymphs, I. hookeri could continuously reproduce throughout the year in Western Kenya.

THE EFFECT OF LIVESTOCK GRAZING ON THE RAINBOW GRASSHOPPER: POPULATION DIFFERENCES AND ECOLOGICAL CORRELATES

This study examined the effect of livestock grazing on a common herbivore in semiarid grasslands of the Southwest: the rainbow grasshopper, Dactylotum variegatum. Population attributes and key environmental variables were compared between sites on active cattle ranches and sites on a 3160-ha ungrazed sanctuary. Although density of D. variegatum nymphs did not differ significantly between grazed and ungrazed sites, adult density was significantly lower on grazed sites, and sex ratios differed between grazed and ungrazed sites over time. Grazed sites had higher percentages of bare ground and fewer Baccharis pteronioides, a common shrub. However, only bare ground was highly correlated with adult D. variegatum density; lower adult densities were associated with more bare ground. At a smaller scale, D. variegatum tended to be more abundant in areas with B. pteronioides on grazed sites and less abundant in areas on un-grazed sites. This study suggests that cattle grazing influences ecological variables important in determining density, sex ratio, and spatial distribution of D. variegatum at both large and small spatial scales.

Visual Evaluation and Recognition of Hosts by the Tick Parasitoid, Ixodiphagus hookeri (Hymenoptera: Encyrtidae)

The role of host size, movement, feeding status, color, and species in the visual host evaluation and recognition behavior of the tick parasitoid, Ixodiphagus hookeri Howard was investigated. Freshly emerged female parasitoids were subjected to a choice bioassay, where the test materials were placed in sealed vials and the vials placed in a Petri dish. When presented with A. variegatum live and mummified nymphs, females examined: larger nymphs significantly longer than smaller nymphs, fed nymphs significantly longer than unfed nymphs, dead and live nymphs for similar lengths of time, and grey live nymphs and yellow-brown and dark brown mummified nymphs for similar lengths of time. The total number of visits to vials containing these test materials were also not significantly different, except there were significantly more visits to yellow-brown mummies when compared to the number of visits to dark brown mummies. When presented with A. variegatum (host) and R. appendiculatus (nonhost) nymphs, the females examined A. variegatum nymphs significantly longer than R. appendiculatus nymphs. The total number of visits to vials containing A. variegatum nymphs were significantly more than the visits to the vials containing R. appendicualtus nymphs. Furthermore, females spent significantly more total examination time per visit on larger and fed A. variegatum nymphs when compared to smaller and unfed nymphs, respectively. Direct and indirect detection were significant when females were presented with fed versus unfed A. variegatum nymphs, grey nymphs versus yellow-brown mummies, and R. appendiculatus versus A. variegatum nymphs. Direct and indirect detection for the rest of the bioassays were not significantly different. Finally, The percentages of females contacting large fed A. variegatum nymphs first were significantly different from those of females contacting small unfed R. appendiculatus nymphs first. The firstcontact percentages for the rest of the bioassays were not significantly different.

Cattle and Amblyomma variegatum Odors Used in Host Habitat and Host Finding by the Tick Parasitoid, Ixodiphagus hookeri

The response of mated naive Ixodiphagus hookeri females to cattle and Amblyomma variegatum nymphal odors was tested in a Y-tube olfactometer. I. hookeri females were attracted to cattle urine, dung, and odors from tick-free feeding sites of A. variegatum nymphs on cattle, e.g., dewlaps, front heels, and hind heels. Tick-free scrotal odors did not attract the parasitoids. Furthermore, odors from off-host unfed and fed A. variegatum nymphs did not attract the parasitoids, despite an increase in the number of the nymphs to amplify any odor signal. A blend of odors from feeding on-host nymphs and cattle scrota attracted the parasitoids. In T-tube bioassays, I. hookeri females were attracted to hexane washes and fecal extracts of A. variegatum nymphs.

Parasitism of Amblyomma variegatum by a Hymenopteran Parasitoid in the Laboratory, and Some Aspects of Its Basic Biology

Abstract Unfed Amblyomma variegatum F. nymphs were experimentally infected in the laboratory with a hymenopteran parasitoid, Ixodiphagus hookeri (Howard). The parasitoid was originally obtained from ticks collected from cattle in the Trans-Mara area of Kenya where it naturally infects 50% of A. variegatum nymphs. In the field, nymphs collected from cattle were found to be infected, but not those collected from grass. The optimum temperature for emergence of parasitoids from the nymphs in the laboratory was 28°C, but emerged parasitoids were more active and survived longer at 22°C. There was a higher proportion of parasitoid nonemergence from laboratory-infected than from the field-infected nymphs. This study is the first record of infection of A. variegatum in the laboratory with a parasitoid. Possibilities of mass rearing and use of the parasitoid as a biocontrol agent for ticks are discussed.

Wildlife as Hosts for Ticks (Acari) in Antigua, West Indies

A survey was conducted to determine the status of wild mammals and birds as hosts for Amblyomma variegatum (F.) and other tick species in Antigua. Surveys of wild mammals and birds were conducted periodically from September 1988 through May 1991. Wild mammals surveyed included the small Indian mongoose (Herpestes auropunctatus Hodgson), Norway rat (Rattus norvegicus Berkenhout), and house mouse (Mus musculus L.), but only mongooses were surveyed intensively. Larvae and nymphs of A. variegatum, larvae of Boophilus microplus (Canestrini), and larvae of Ornithodoros puertoricensis (Fox) were recovered. The mean prevalences of infestation of mongooses by A. variegatum larvae and nymphs were 4.7 and 1.3%, respectively; maximums were 16.1 and 5.0%, respectively. The mean prevalence of infestation of mongooses by B. microplus was 3.2%. O. puertoricensis is reported from Antigua for the first time. The mean prevalence of infestation of mongooses by O. puertoricensis larvae was 41.2%, but seasonal prevalences ranged from 27.8 to 55.0%. Of 610 birds representing 16 species, three Carib grackles (Quiscalus lugubris Swainson) were each infested with one larva of A. variegatum.

The local effect of hypersensitive or inflammatory reactions to nymphal Amblyomma variegatum on simultaneous infections with Dermatophilus congolensis

Amblyomma variegatum nymphs were applied to sites infected with Dermatophilus congolensis on eight rabbits. Four rabbits were previously sensitized to the feeding of nymphal A. variegatum to produce hypersensitive reactions to the tick feeding; the remaining four rabbits had no previous exposure to nymphal A. variegatum and produced inflammatory reactions to the tick feeding. The resulting dermatophilosis infections were assessed for three weeks and there was a correlation between the position of the inflammatory tick attachment sites and the foci of infection. There was a significant increase in the lesions at sites with inflammatory reactions to the ticks, compared with sites not exposed to tick feeding; these differences appeared to be due to individual variation in the host response and were not sustained throughout the assessment.

Le portage asymptomatique de bovins et chèvres Créole guéris de la cowdriose en Guadeloupe

Creole goats and cattle in Guadeloupe can be carriers of cowdriosis (heartwater: Cowdria ruminantium) after recovery for a period as long as 11 months in goats and 2 months in cattle. The carrier status was demonstrated by feeding Amblyomma variegatum nymphs on recovered animals and the resulting adult ticks on susceptible goats. Cowdria ruminantium was not detected permanently during the carrier status.

A cloned DNA probe identifies Cowdria ruminantium in Amblyomma variegatum ticks

Heartwater, caused by Cowdria ruminantium and transmitted by ticks of the genus Amblyomma, is a constraint to ruminant animal production in sub-Saharan Africa. This rickettsial disease could spread from endemically infected areas of sub-Saharan Africa and certain Caribbean islands to other countries, including the United States, in which Amblyomma ticks exist. To detect C. ruminantium in tick vectors and animals, we made DNA probes from C. ruminantium DNA isolated from endothelial cell cultures. Two clones were evaluated; pCS20 from Crystal Springs (Zimbabwe) strain DNA had a 1,306-bp insert, and pCR9 from Kiswani (Kenya) strain DNA had a 754-bp insert. Both DNA probes detected 1 ng of Crystal Springs DNA; however, the pCS20 probe had a 10-fold-greater ability to discriminate between C. ruminantium DNA and DNA from other organisms. Also, the pCS20 probe did not hybridize to 400 ng (highest amount tested) of DNA from bovine cells, 3 protozoa, 3 rickettsiae, and 12 bacteria. In all experiments, C. ruminantium DNA was detected in midguts from 99 of 160 Amblyomma variegatum nymphs infected as larvae and in midguts from 38 of 80 adult ticks infected as nymphs but not in midguts from control nymphs and adults. The presence of C. ruminantium in nymphs and adults was confirmed by transmission of heartwater to goats. The DNA sequences of both probes were determined; synthetic oligonucleotides from pCS20 are recommended as DNA probes for C. ruminantium.

Histological Reactions to Bites of Amblyomma variagatum and Rhipicephalus appendiculatus (Acari: Ixodidae) Fed Simultaneously on Naive or Sensitized Rabbits

Histopathological studies on attachment sites of nymphs, 48 h after attachment on three groups of rabbits, have revealed differences that were related to the rabbits' previous tick experience. Feeding lesions caused by Amblyomma variegatum (F.) in tick-naive rabbits were extensive and the total number of inflammatory cells was about 10 times greater than that in the feeding lesions caused by Rhipicephalus appendiculatus Neumann fed simultaneously on contralateral ears. Rabbits that were previously sensitized either to A. variegatum or R. appendiculatus by repeated tick infestations showed epidermal vesiculation and significant mobilization of eosinophils at the homologous tick feeding sites, events that did not occur with tick-naive rabbits. The feeding of A. variegatum nymphs on rabbits sensitized to R. appendiculatus produced a similar type of intense reaction, but the cellular responses to R. appendiculatus in the skin of rabbits sensitized to the A. variegatum were negligible.

Difference in vector competence of two species of sympatric ticks, Amblyomma variegatum and Rhipicephalus appendiculatus, for Dugbe virus (Nairovirus, Bunyaviridae)

Amblyomma variegatum was shown to be a competent vector of Dugbe (DUG) virus whereas Rhipicephalus appendiculatus was not. When DUG virus was taken up orally by A. variegatum nymphs, during capillary feeding, the virus replicated and persisted through moulting to the following adult stage. In contrast, although DUG virus replicated in capillary fed R. appendiculatus nymphs, the virus did not persist trans-stadially into the adult stage. If the oral route of infection was by-passed by direct inoculation into the haemocoel, DUG virus replicated and survived trans-stadially in both tick species, and was subsequently transmitted during feeding. The different responses of R. appendiculatus to oral and intra-coelomic routes suggest that DUG virus is able to infect the gut of this tick species, but that release of the virus from the gut is inhibited.

Monitoring of naturally acquired and artificially induced immunity to Amblyomma variegatum and Rhipicephalus appendiculatus ticks under field and laboratory conditions.

The ability of rabbits, goats and cattle to acquire immunity to the ixodid ticks Amblyomma variegatum and Rhipicephalus appendiculatus was studied under laboratory and field conditions. Rabbits were successfully immunized with crude salivary gland extract (SGE) and midgut extract (ME) obtained from flat or partly fed female R. appendiculatus ticks. The lowest numbers of larvae were produced by females fed on rabbits immunized with unfed midgut extract. Similar reductions in larval production could be induced after three infestations of rabbits with adult R. appendiculatus. Also, successive feedings of nymphs of R. appendiculatus on rabbits resulted in significantly reduced engorgement weights. Skin testing with SGE induced delayed-type hypersensitivity reactions, which could be correlated with immunity to R. appendiculatus in rabbits. Moreover, circulating antibodies were detected in rabbits with an ELISA using SGE of R. appendiculatus. Immunity to A. variegatum nymphs could be induced in rabbits by repeated infestations, but this failed in goats. Immunization of goats with midgut extract from adult A. variegatum did not protect against subsequent nymphal challenge, but strong skin reactions were noticed when adults ticks fed on immunized goats. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of SGE and ME from A. variegatum revealed the presence of 48 protein bands in SGE and 29 bands in midgut extract. Western blotting employing serum from a rabbit immune to R. appendiculatus recognized a number of bands in SGE from R. appendiculatus, but also in SGE of A. variegatum. Immunity acquired by cattle to ixodid tick infestations under field conditions was monitored by skin testing with SGE and western blot analysis. In general, cattle with the lowest tick numbers manifested the strongest delayed-type hypersensitivity responses. Finally, western blot analysis employing sera from tick-infested and tick-naive cattle could not be related to actual immune status.

Vector capacity of Rhipicephalus appendiculatus and Amblyomma variegatum for Thogoto and Dhori viruses

Experimental studies were undertaken to ascertain the vector potential of Rhipicephalus appendiculatus Neumann and Amblyomma variegatum Fabricius for Thogoto (THO) and Dhori (DHO) viruses, candidate members of the Orthomyxoviridae. In the first set of experiments, ticks were infected orally by feeding on viraemic hamsters. THO virus replicated in R. appendiculatus and A. variegatum, persisted trans-stadially in both tick species, and was transmitted to susceptible hosts during feeding. In contrast, both R. appendiculatus and A. variegatum were refractory to per os infection by DHO virus. In the second set of experiments, engorged R. appendiculatus and A. variegatum nymphs were parenterally inoculated with DHO virus. The virus persisted trans-stadially in both tick species and was transmitted by bite to susceptible hosts. These results indicate that the midgut acts as a barrier to per os infection of R. appendiculatus and A. variegatum by DHO virus. However, when this barrier is bypassed, i.e. by parenteral inoculation, both R. appendiculatus and A. variegatum can serve as efficient vectors of DHO virus.