Appendiculatus Ticks Research Articles

Codon optimization of voraxin α sequence enhances the immunogenicity of a recombinant vaccine against Hyalomma anatolicum infestation in rabbits

Research has shown that voraxin α derived from male ticks stimulates blood feeding to engorge in female ticks. Whereas, the oviposition rate, egg weight, and body weight of female ticks were reduced in animals vaccinated with recombinant (r-) voraxin α. These data suggest a potential role of r-voraxin α as a functional anti-tick antigen in Rhipicephalus appendiculatus and Amblyomma hebraeum tick infestation. This study investigated the immunogenicity of r-voraxin α protein from Hyalomma anatolicum (H. anatolicum) tick as an anti-tick vaccine in rabbits. The H. anatolicum voraxin α sequence was optimized according to the codon usage in E. coli before being sub-cloned into pQE30. The gene sequence of the voraxin α was synthesized, verified by DNA sequencing, cloned in a pQE30 vector, and transformed into E. coli. Then, the expression of the r-voraxin α protein was confirmed by SDS-PAGE and Western blot analysis. Subsequently, three rabbits were immunized with the r-voraxin α as the vaccinated group, whereas three rabbits without injection were considered the control group. The result indicated the success of cloning of codon-optimized H. anatolicum voraxin α gene. Moreover, the expression of the r-voraxin α protein (approximately 18 kDa) in the bacterial expression system was confirmed by SDS-PAGE and Western blot analysis. The results of this study showed that the mortality rate in vaccine recipients increased compared to the control group (P < 0.01). Also, the egg weight, oviposition rate, and engorgement weight of female ticks fed from vaccinated animals were significantly reduced compared to the control group (P < 0.01). The results confirmed that the codon-optimized H. anatolicum voraxin α gene expressed in the bacterial expression system could be a suitable anti-tick vaccine against H. anatolicum tick infestation.

A longitudinal study of tick infestations and management practices in young stock smallholder dairy cattle in the Tanga region, Tanzania

A longitudinal study involving cohorts of young stock animals (&lt;3 years old) kept on smallholder dairy farms in the Tanga region, Tanzania, was conducted to identify and quantify tick infestations and describe on-farm tick management practices. A total of 549 young stock were followed for 23 months, and 3127 grazing and acaricide treatment observations were obtained. Tick counts were low in the present study, and while more than 30% of animals had a tick on one occasion, most animals had no ticks at most examinations. The most commonly encountered spp. were R. appendiculatus, Boophilus spp., A. variegatum and R. evertsi. Most animals were zero-grazed, a management system that significantly limits animal exposure to ticks; however, grazed calves were more than three times more likely to carry R. appendiclatus and Boophilus spp. ticks than zero-grazed calves were (OR [odds ratio] = 4.4, P &lt; 0.001; 3.6, P &lt; 0.001). There were no significant differences among sampling rounds in the numbers of animals carrying R. appendiculatus ticks (likelihood ratio test P = 0.520). Hand spraying was the common method used, acaricides that require on farm dilution were most commonly used, and most farmers used this method at concentrations considerably less than those recommended by the manufacturer, most likely due to poor understanding, high cost (tempting them to over dilute) and weak extension/veterinary services. Variation in acaricide treatment frequency was not associated with variations in tick attachment, suggesting that most farms were under a strong acaricide regime or at irregular intervals or with tick-building resistance. Our findings suggest that geographic variations in vector densities might partly influence the extent of infection caused by tick-borne pathogens. The applicability of this study to large and extensive cattle production systems is recommended to widen the scope of tick management practices while generating additional data that will inform national tick and tick-borne disease control policy reform in Tanzania.

Assessment of an In Vitro Tick Feeding System for the Successful Feeding of Adult Rhipicephalus appendiculatus Ticks

This study assessed the efficiency of a new in vitro tick feeding system for the adult Rhipicephalus appendiculatus tick and compared the impact of different blood anticoagulating factors on their feeding process. A total of 10 feeders were each seeded with 30 or 60 R. appendiculatus adults. Bovine blood was added into each unit and changed every 12 h for 4 to 10 days during which tick attachment and engorgement was assessed. The tick attachment observed 4 days after feeding was 80.0% (48/60), 75.8% (182/240), and 70.8% (170/240) for lithium heparin, citrate phosphate dextrose, and defibrinated blood, respectively, with no significant difference (p &gt; 0.05) between the anticoagulants used. However, the ticks fed on heparinized and defibrinated blood reached repletion status. The in vitro tick feeding system was successfully used to feed adult R. appendiculatus ticks until repletion. This system could be used to facilitate studies on tick-pathogen interactions, such as those involved in the East Coast fever disease.

A cross-sectional survey to establish Theileria parva prevalence and vector control at the wildlife-livestock interface, Northern Tanzania

East Coast fever (ECF) in cattle is caused by the protozoan parasite Theileria parva, transmitted by Rhipicephalus appendiculatus ticks. In cattle ECF is often fatal, causing annual losses >$500 million across its range. The African buffalo (Syncerus caffer) is the natural host for T. parva but the transmission dynamics between wild hosts and livestock are poorly understood. This study aimed to determine the prevalence of T. parva in cattle, in a 30 km zone adjacent to the Serengeti National Park, Tanzania where livestock and buffalo co-exist, and to ascertain how livestock keepers controlled ECF and other vector-borne diseases of cattle.A randomised cross-sectional cattle survey and questionnaire of vector control practices were conducted. Blood samples were collected from 770 cattle from 48 herds and analysed by PCR to establish T. parva prevalence. Half body tick counts were recorded on every animal. Farmers were interviewed (n = 120; including the blood sampled herds) using a standardised questionnaire to obtain data on vector control practices. Local workshops were held to discuss findings and validate results.Overall prevalence of T. parva in cattle was 5.07% (CI: 3.70−7.00%), with significantly higher prevalence in older animals. Although all farmers reported seeing ticks on their cattle, tick counts were very low with 78% cattle having none. Questionnaire analysis indicated significant acaricide use with 79% and 41% of farmers reporting spraying or dipping with cypermethrin-based insecticides, respectively. Some farmers reported very frequent spraying, as often as every four days. However, doses per animal were often insufficient.These data indicate high levels of acaricide use, which may be responsible for the low observed tick burdens and low ECF prevalence. This vector control is farmer-led and aimed at both tick- and tsetse-borne diseases of livestock. The levels of acaricide use raise concerns regarding sustainability; resistance development is a risk, particularly in ticks. Integrating vaccination as part of this community-based disease control may alleviate acaricide dependence, but increased understanding of the Theileria strains circulating in wildlife-livestock interface areas is required to establish the potential benefits of vaccination.

Review of diagnostic procedures and control of some viral diseases causing abortion and infertility in small ruminants in Egypt

Sheep and goats represent an essential source of meat, milk and wool production. Infectious viral diseases of small ruminants hinder the expected benefits from these animals. The aim of the present review was to shed light on diagnostic procedures and monitoring of some important infectious viral diseases that affect small ruminants. Border disease (BDV) is caused by Pestiviruses. The affected herds are usually expressed high rates of infertility and production of underweighted-lambs. Affected lambs usually die within days after lambing. A commercial killed whole virus vaccine was produced for BDV. Louping ill (LI) is caused by the Louping ill virus, which is transmitted by Ixods ricinus ticks. The LI virus typically causes fever, anorexia and encephalomyelitis. Death could occur 1-3 days after the beginning of signs. The available Louping ill vaccine is composed of inactivated killed virus. Nairobi sheep disease (NSD) is a tick-born infectious disease caused by Rhipicephalus appendiculatus ticks, characterized clinically by fever, abortion hemorrhagic gastroenteritis and high mortality. Inactivated killed oil adjuvant virus vaccines are available for the control of Nairobi sheep disease. Akabane disease (AKAV) is a Culicoides borne viral disease belongs to orthobunyavirus that has a teratogenic effect on the fetus of cattle and small ruminants. A live attenuated virus vaccine and inactivated virus are commonly used. This review concluded that the modern diagnostic tools are urgently needed not only for diagnosis but also for monitoring viral disease control and control programs.

First detection of Theileria parva in cattle from Cameroon in the absence of the main tick vector Rhipicephalus appendiculatus.

A major risk factor for the spread of livestock diseases and their vectors is the uncontrolled transboundary movement of live animals for trade and grazing. Such movements constrain effective control of tick‐transmitted pathogens, including Theileria parva. Only limited studies have been undertaken to identify ticks and tick‐borne diseases (TTBDs) affecting cattle in central African countries, including Cameroon. We hereby report the collection of baseline data on the prevalence of T. parva in Cameroon through a countrywide cross‐sectional survey, conducted in 2016, involving collection of blood samples from cattle from 63 sites across the five agro‐ecological zones (AEZs) of the country. ELISA‐based surveillance of infected cattle was performed on 479 randomly selected samples and revealed specific antibodies to T. parva in 22.7% and T. mutans in 41.1% of cattle. Screening of 1,340 representative DNA samples for the presence of T. parva identified 25 (1.86%) positives using a p104 antigen gene‐based nested PCR assay. The positives were distributed across agro‐ecological zones I, II, III and V. None of the p104 positive cattle exhibited clinical symptoms of East Coast fever (ECF). Using reverse line blot (RLB), 58 (4.3%) and 1,139 (85%) of the samples reacted with the T. parva and T. mutans oligonucleotide probes, respectively. This represents the first report of T. parva from Cameroon. Surprisingly, no Rhipicephalus appendiculatus ticks, the main vector of T. parva, were identified in a parallel study involving comprehensive morphological and molecular survey of tick species present in the country. Only two of the 25 p104 positive cattle were PCR‐positive for the CD8+ T‐cell target schizont‐expressed antigen gene Tp1. Cloning and sequencing of Tp1 amplicons revealed sequence identity with the reference T. parva Muguga. This new finding raises serious concerns of a potential spread of ECF into the central African region.

Rhipicephalus appendiculatus ticks transmit Theileria parva from persistently infected cattle in the absence of detectable parasitemia: implications for East Coast fever epidemiology

BackgroundEast Coast fever (ECF) is a devastating disease of cattle and a significant constraint to improvement of livestock production in sub-Saharan Africa. The protozoan parasite causing ECF, Theileria parva, undergoes obligate sexual stage development in its tick vector Rhipicephalus appendiculatus. Tick-borne acquisition and transmission occurs transstadially; larval and nymphal ticks acquire infection while feeding and transmit to cattle when they feed after molting to the next stage. Much of the current knowledge relating to tick-borne acquisition and transmission of T. parva has been derived from studies performed during acute infections where parasitemia is high. In contrast, tick-borne transmission during the low-level persistent infections characteristic of endemic transmission cycles is rarely studied.MethodsCattle were infected with one of two stocks of T. parva (Muguga or Marikebuni). Four months post-infection when parasites were no longer detectable in peripheral blood by PCR, 500 R. appendiculatus nymphs were fed to repletion on each of the cattle. After they molted to the adult stage, 20 or 200 ticks, respectively, were fed on two naïve cattle for each of the parasite stocks. After adult ticks fed to repletion, cattle were tested for T. parva infection by nested PCR and dot blot hybridization.ResultsOnce they had molted to adults the ticks that had fed as nymphs on Muguga and Marikebuni infected cattle successfully transmitted Theileria parva to all naïve cattle, even though T. parva infection was not detectable by nested PCR on salivary gland genomic DNA of a sample of individual ticks. However, a salivary gland homogenate from a single Marikebuni infected tick was able to infect primary bovine lymphocytes. Infection was detected by nested p104 PCR in 3 of 4 calves and detected in all 4 calves by T. parva 18S nested PCR/dot blot hybridization.ConclusionWe show that R. appendiculatus ticks are able to acquire T. parva parasites from infected cattle even in the absence of detectable parasitemia. Although infection was undetectable in a sample of individual ticks, cumulatively as few as 20 ticks were able to transmit T. parva to naïve cattle. These results have important implications for our understanding of T. parva transmission by R. appendiculatus in ECF endemic regions.

Essential oils of indigenous plants protect livestock from infestations of Rhipicephalus appendiculatus and other tick species in herds grazing in natural pastures in western Kenya

The effects of formulated essential oils of Tagetes minuta and Tithonia diversifolia on Rhipicephalus appendiculatus infesting livestock were evaluated in semi-field experiments. Forty-five zebu cattle naturally infested with ticks were randomly selected from 15 herds, three animals from each. Of the three animals within each herd, one was treated with 1 g of petroleum jelly (control), one with 1 g of essential oil of T. minuta and one with 1 g of essential oil of T. diversifolia on the inner side of ear pinna, the preferred feeding site of R. appendiculatus. Tick infestation on each treated host animal was monitored daily for 18 days by counting the number of ticks attached to the animals. Within 1–4 days post-treatment, the number of ticks on animals treated with essential oils was reduced by more than half the original population. By the 5th day post-treatment, more than 75 and 60% of R. appendiculatus and other tick species, respectively, became dislodged and dropped off. A stronger repellent effect was shown by the essential oil of T. minuta than the essential oil of T. diversifolia. Mean residual protection afforded by T. minuta was 12.5 days and for T. diversifolia 7.9 days. There was no significant difference in the effectiveness of essential oils between male and female R. appendiculatus. Both T. minuta and T. diversifolia essential oils affected several other less dominant but economically important tick species. Results suggest the potential for essential oil formulations in integrated pest management of ticks and associated tick-borne diseases among resource-limited livestock farmers in tropical Africa.

Analyses of mitochondrial genes reveal two sympatric but genetically divergent lineages of Rhipicephalus appendiculatus in Kenya.

BackgroundThe ixodid tick Rhipicephalus appendiculatus transmits the apicomplexan protozoan parasite Theileria parva, which causes East coast fever (ECF), the most economically important cattle disease in eastern and southern Africa. Recent analysis of micro- and minisatellite markers showed an absence of geographical and host-associated genetic sub-structuring amongst field populations of R. appendiculatus in Kenya. To assess further the phylogenetic relationships between field and laboratory R. appendiculatus tick isolates, this study examined sequence variations at two mitochondrial genes, cytochrome c oxidase subunit I (COI) and 12S ribosomal RNA (rRNA), and the nuclear encoded ribosomal internal transcribed spacer 2 (ITS2) of the rRNA gene, respectively.ResultsThe analysis of 332 COI sequences revealed 30 polymorphic sites, which defined 28 haplotypes that were separated into two distinct haplogroups (A and B). Inclusion of previously published haplotypes in our analysis revealed a high degree of phylogenetic complexity never reported before in haplogroup A. Neither haplogroup however, showed any clustering pattern related to either the geographical sampling location, the type of tick sampled (laboratory stocks vs field populations) or the mammalian host species. This finding was supported by the results obtained from the analysis of 12S rDNA sequences. Analysis of molecular variance (AMOVA) indicated that 90.8 % of the total genetic variation was explained by the two haplogroups, providing further support for their genetic divergence. These results were, however, not replicated by the nuclear transcribed ITS2 sequences likely because of recombination between the nuclear genomes maintaining a high level of genetic sequence conservation.ConclusionsCOI and 12S rDNA are better markers than ITS2 for studying intraspecific diversity. Based on these genes, two major genetic groups of R. appendiculatus that have gone through a demographic expansion exist in Kenya. The two groups show no phylogeographic structure or correlation with the type of host species from which the ticks were collected, nor to the evolutionary and breeding history of the species. The two lineages may have a wide geographic distribution range in eastern and southern Africa. The findings of this study may have implications for the spread and control of R. appendiculatus, and indirectly, on the transmission dynamics of ECF.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1631-1) contains supplementary material, which is available to authorized users.

English

This study was carried out in Fangak County, Jonglei State, South Sudan, with aim to identify the main ticks species and follow north limit of the theileriosis in Jonglei. Three localities namely; Hai, Toggar and Bichoul kun village in Phoum payam were selected as suspected area for East Coast fever (ECF), Group A) and Kuer kan in Manjang payams were selected as non suspected area for ECF (Group B). These groups A and B were based on animal movement, trade businessand intermarriages (animals paid for dowry). A total of 120 sera were collected from cattle of different age groups. The serum samples were tested using indirect polymorphic immunodominant molecule (PIM) ELISA to detect Theileria parva antibodies. The results indicated that 5/44 (11.4%) samples from non suspected area (group B) and 48/76 (63.2%) samples from suspected area (Group A) revealed antibodies. The overall positivity was 53/120 (44.17%) which was highly significant (P < 0.001) according to the locations. Three tick genera were recorded, Amblyomma, Hyalomma and Rhipicephalus. The species were A. variegatum, A. lepidum, H. rufipes, R. (B.) decoloratus, R. (B.) annulatus, R. e. evertsi and R. sanguineus. The most abundant tick species was A. variegatum, constituting 62%, while the lowest tick recorded was R. e. evertsi with prevalence rate of 2%. No R. appendiculatus tick was seen in Fangak area; while T. parva antibodies were detected. Regarding the fact that T. parva antibodies were present in the area with 44.17% prevalence, more efforts are needed to determine the extension of ECF and its vector R. appendiculatus to the northern parts of Jonglei State and this result of ECF antibodies could be an alarm to migrate cattle owners from South Sudan. Key words: Ticks, east coast fever (ECF), ELISA, South Sudan.

Efficacy of spot-spray application of Metarhizium anisopliae formulated in emulsifiable extract of Calpurnia aurea in attracting and infecting adult Rhipicephalus appendiculatus ticks in semi-field experiments

The efficacy of spot-spray application of conidia of Metarhizium anisopliae (Metsch.) Sorok. conidia formulated in an emulsifiable extract of Calpurnia aurea (Ait.) Benth in attracting and infecting Rhipicephalus appendiculatus Neumann, 1901 ticks was evaluated in semi-field experiments. Formulation was applied on a 900-cm² spot and ticks released from various distances. Attracted ticks were individually placed in glass vials to determine the number of M. anisopliae conidia picked by a single tick. Infected ticks were either transferred in glass vials or maintained in the laboratory until death or exposed to rabbits in order to investigate the effects of fungal infection on feeding and potential reproduction parameters. Data showed that individual ticks picked between 3.2 and 4.1 × 105 conidia. Mortality of 83 % was observed among ticks maintained in the laboratory conditions. The engorgement period of fungus-infected ticks significantly increased by 16 % compared to the control. A significant reduction (P < 0.0001) was recorded in body weight, egg-mass and egg hatchability from fungus-infected females. It is therefore possible to use M. anisopliae formulated in C. aurea extract for an integrated pest management approach. This study showed that conidia of M. anisopliae formulated in C. aurea extract are effective in attracting, infecting and causing mortalities of R. appendiculatus ticks. Moreover, the reproduction performance of fungus-infected females was significantly reduced.

Demonstration of lumpy skin disease virus infection in Amblyomma hebraeum and Rhipicephalus appendiculatus ticks using immunohistochemistry

Lumpy skin disease (LSD) is caused by lumpy skin disease virus (LSDV), a member of the genus Capripoxvirus. Transmission of the virus has been associated with haematophagous insects such as Stomoxys calcitrans as well as Aedes and Culex species of mosquitoes. Recent studies have reported the transmission of the virus by Amblyomma hebraeum, Rhipicephalus appendiculatus, and Rhipicephalus decoloratus ticks and the presence of LSDV in saliva of A. hebraeum and R. appendiculatus ticks. The aim of this study was to determine which tick organs become infected by LSDV following intrastadial infection and transstadial persistence of the virus in A. hebraeum and R. appendiculatus ticks. Nymphal and adult ticks were orally infected by feeding them on LSDV-infected cattle. Partially fed adult ticks were processed for testing while nymphs were fed to repletion and allowed to moult to adults before being processed for testing. The infection in tick organs was determined by testing for the presence of the viral antigen using monoclonal antibodies with immunohistochemical staining. The viral antigen was detected in salivary glands, haemocytes, synganglia, ovaries, testes, fat bodies, and midgut. Since the virus was shown to be able to cross the midgut wall and infect various tick organs, this may indicate potential for biological development and transmission of LSDV in ticks. This study strengthens the previously reported evidence of the occurrence of LSDV in tick saliva.

Novel Immunomodulators from Hard Ticks Selectively Reprogramme Human Dendritic Cell Responses

Hard ticks subvert the immune responses of their vertebrate hosts in order to feed for much longer periods than other blood-feeding ectoparasites; this may be one reason why they transmit perhaps the greatest diversity of pathogens of any arthropod vector. Tick-induced immunomodulation is mediated by salivary components, some of which neutralise elements of innate immunity or inhibit the development of adaptive immunity. As dendritic cells (DC) trigger and help to regulate adaptive immunity, they are an ideal target for immunomodulation. However, previously described immunoactive components of tick saliva are either highly promiscuous in their cellular and molecular targets or have limited effects on DC. Here we address the question of whether the largest and globally most important group of ticks (the ixodid metastriates) produce salivary molecules that specifically modulate DC activity. We used chromatography to isolate a salivary gland protein (Japanin) from Rhipicephalus appendiculatus ticks. Japanin was cloned, and recombinant protein was produced in a baculoviral expression system. We found that Japanin specifically reprogrammes DC responses to a wide variety of stimuli in vitro, radically altering their expression of co-stimulatory and co-inhibitory transmembrane molecules (measured by flow cytometry) and their secretion of pro-inflammatory, anti-inflammatory and T cell polarising cytokines (assessed by Luminex multiplex assays); it also inhibits the differentiation of DC from monocytes. Sequence alignments and enzymatic deglycosylation revealed Japanin to be a 17.7 kDa, N-glycosylated lipocalin. Using molecular cloning and database searches, we have identified a group of homologous proteins in R. appendiculatus and related species, three of which we have expressed and shown to possess DC-modulatory activity. All data were obtained using DC generated from at least four human blood donors, with rigorous statistical analysis. Our results suggest a previously unknown mechanism for parasite-induced subversion of adaptive immunity, one which may also facilitate pathogen transmission.

Detection of lumpy skin disease virus in saliva of ticks fed on lumpy skin disease virus-infected cattle.

Lumpy skin disease is an economically important disease of cattle that is caused by the lumpy skin disease virus (LSDV), which belongs to the genus Capripoxvirus. It is endemic in Africa and outbreaks have also been reported in the Middle-East. Transmission has mostly been associated with blood-feeding insects but recently, the authors have demonstrated mechanical transmission by Rhipicephalus appendiculatus as well as mechanical/intrastadial and transstadial transmission by Amblyomma hebraeum. Saliva is the medium of transmission of pathogens transmitted by biting arthropods and, simultaneously, it potentiates infection in the vertebrate host. This study aimed to detect LSDV in saliva of A. hebraeum and R. appendiculatus adult ticks fed, as nymphs or as adults, on LSDV-infected animals, thereby also demonstrating transstadial or mechanical/intrastadial passage of the virus in these ticks. Saliva samples were tested for LSDV by real-time PCR and virus isolation. Supernatants obtained from virus isolation were further tested by real-time PCR to confirm that the cytopathic effects observed were due to LSDV. Lumpy skin disease virus was detected, for the first time, in saliva samples of both A. hebraeum and R. appendiculatus ticks. At the same time, mechanical/intrastadial and transstadial passage of the virus was demonstrated and confirmed in R. appendiculatus and A. hebraeum.

Identification of hard ticks (Acari: Ixodidae) and seroprevalence to Theileria parva in cattle raised in North Kivu Province, Democratic Republic of Congo

This study aimed to identify tick species and to determine their relationship with the Theileria parva seroprevalence in cattle raised under an extensive farming system in North Kivu Province, Democratic Republic of Congo in two agro-ecological zones namely medium (1,000-1,850 m) and high (>1,850 m) altitude. Among the 3,215 ticks collected on 482 animals, from February to April 2009, Rhipicephalus appendiculatus (64.26 %), the main vector of T. parva, was the most abundant species followed by Rhipicephalus decoloratus (35.49 %) and Amblyomma variegatum (0.25 %). The mean burden of R. appendiculatus tick per infested animal appeared significantly higher at medium (6.5 ± 0.22 ticks) than at high (0.07 ± 0.3 ticks) altitude (P < 0.05). However, an indirect fluorescent antibody test carried out on 450 blood samples revealed a global T. parva seroprevalence of 43 % (95 % CI: 38-47) which was not significantly (P > 0.05) different between medium (48.4 %; 95 % CI: 38-49) and high (41.9 %; 95 % CI: 35-49) altitude. These relatively low seroprevalences suggest that there is a state of endemicity to T. parva infection in the study area. The presence of the tick vector on animals was associated with an increased risk of being seropositive to T. parva infection (odds ratio = 2.04; 95 % CI: 1.8-2.3; P < 0.001). The results suggest the need for a longitudinal study to investigate the seasonal dynamics of tick species and T. parva infection. The rate of tick infection should also be evaluated in order to determine the intensity of T. parva transmission to cattle.

Population attributable fractions of farm vector tick (Rhipicephalus appendiculatus) presence on Theileria parva infection seroprevalence under endemic instability

The primary objective of this study was to assess the impact of Rhipicephalus appendiculatus tick presence (exposure variable) on Theileria parva infection seroprevalence (outcome variable) in a group of cattle belonging to a farm using population attributable fractions (PAF). The analyses were based on a representative sample of 80 traditional smallholder mixed farms. The farms were selected by first stratifying the population administratively and implementing a multistage random sampling in Mbeere district in Kenya. The PAFs were estimated using the stratified, Bruzzi, and sequential partitioned PAF approaches. A secondary objective was, thus, to evaluate the impact of the approaches on the PAF estimates. The stratified and Bruzzi approaches estimated proportion of T. parva infection cases directly attributable to the exposure after controlling for confounding by agro-ecological zone (AEZ). The sequential partitioned PAF approach estimated a PAF associated with exposure after adjusting for any effect that the AEZ may have had by influencing the prevalence of the exposure. All analyses were carried out at the farm level where a farm was classified as infested if the tick was found on cattle on a farm, and infected if at least one animal on a farm was positive for T. parva antibodies. Variance estimation for PAFs was implemented using ‘delete-a-group’ jackknife re-sampling method. The stratified PAF (26.7% [95% CI: 9.0%, 44.4%]) and Bruzzi PAF (26.4% [95% CI: 9.6%, 43.2%]) were consistent in estimating a relatively low impact of farm vector tick presence with a relatively high level of uncertainty. The partitioned PAF (15.5% [95% CI: 1.5%, 29.6%]) suggested that part of the impacts estimated using the stratified PAF and Bruzzi approaches was driven by AEZ effects. Overall, the results suggested that under endemic instability in Mbeere district, (1) presence of R. appendiculatus was not a good indicator of T. parva infection occurrence on a farm; (2) ecological variation could play a role in determining infection impacts. This study provides a preliminary basis for evaluating the potential value and utility of estimating PAFs for variables amenable to control in tick-borne diseases (TBDs) epidemiological studies.

Recombinant Rhipicephalus appendiculatus gut (Ra86) and salivary gland cement (Trp64) proteins as candidate antigens for inclusion in tick vaccines: protective effects of Ra86 on infestation with adult R. appendiculatus

Recombinant Rhipicephalus appendiculatus gut (Ra86) and salivary gland cement (Trp64) proteins as candidate antigens for inclusion in tick vaccines: protective effects of Ra86 on infestation with adult R. appendiculatus Margaret Saimo1,2,*, David O Odongo3,4,*, Stephen Mwaura3, Just M Vlak1, Anthony J Musoke5, George W Lubega2, Richard P Bishop3, Monique M van Oers11Laboratory of Virology, Wageningen University, Wageningen, The Netherlands; 2School of Veterinary Medicine, Makerere University, Kampala, Uganda; 3International Livestock Research Institute, Nairobi, Kenya; 4School of Biological Sciences, University of Nairobi, Nairobi, Kenya; 5Onderstepoort Veterinary Institute, Onderstepoort, Pretoria, South Africa *These two authors made an equal contribution to this workAbstract: Rhipicephalus appendiculatus gut protein Ra86 (variants Ra85A and Ra92A) and the salivary gland cement protein (Trp64) were expressed in the baculovirus-insect cell system. The recombinant gut proteins expressed as soluble proteins and the recombinant cement protein, as insoluble inclusion bodies, were used to immunize rabbits, which were then challenged with larval, nymphal, and adult stages of R. appendiculatus ticks. High tick mortality (23.3%) occurred on adult ticks that fed on rabbits vaccinated with the gut proteins, compared with 1.9% mortality in ticks that fed on unvaccinated na&iuml;ve control rabbits. The mean weight of engorged female ticks was significantly reduced by 31.5% in rabbits vaccinated with the Ra86 recombinant protein compared with controls, as was egg production. Marked effects on these parameters were also observed in adult ticks as a result from vaccination using Trp64, but these were not statistically significant. For both antigens, there was no demonstrable effect on larval or nymphal ticks. This study demonstrates for the first time the protective efficacy of a homolog of Boophilus microplus Bm86 in reducing tick infestation by the adult stage of the three-host tick R. appendiculatus. The results demonstrate the potential of Ra86 for vaccine development against this tick and for the control of East Coast fever.Keywords: baculovirus, Ra85A, Ra92A, Boophilus decoloratus, Boophilus microplus

Colonization of Grande Comore Island by a lineage of Rhipicephalus appendiculatus ticks

BackgroundUnion of the Comoros suffered a severe East Coast Fever epidemic in 2004. Rhipicephalus appendiculatus was probably involved in pathogen transmission as this competent tick species, although previously absent from Comoros, was sampled on 4 animals on one geographical site during the epidemic. We carried out an entomological survey on all three islands of Union of the Comoros to establish cattle tick species distribution with a special emphasis on R. appendiculatus. We investigated R. appendiculatus intraspecific diversity as this species has been previously shown to be split off into two main cytoplasmic lineages with different ecology, physiology and vectorial competence. This survey also included sampling of live cattle imported from Tanzania to investigate the possibility of tick introduction through animal trade.ResultsOur data show that Comoros cattle are infested with Amblyomma variegatum, Rhipicephalus microplus and R. appendiculatus. This latter species has established throughout Grande Comore but is absent from Anjouan and Moheli. Interestingly, 43 out of the 47 sequenced R. appendiculatus ticks belong to one single highly competent lineage while ticks from the other lineage where only found on imported cattle or on cattle parked at the vicinity of the harbor. At last, 2 ticks identified as R. evertsi, a species so far virtually absent on Comoros, were sampled on imported cattle.ConclusionsThis survey shows that importation of live cattle is clearly a source of vector introduction in Comoros. The wide distribution of one highly competent R. appendiculatus lineage on Grande Comore, together with the absence of this species on the two neighbouring islands is in accordance with the rapid and disastrous spread of East Coast Fever epidemics on Grande Comore Island only. Whether the other R. appendiculatus lineage as well as R. evertsi species will succeed in establishing permanently on Grande Comore needs to be monitored.

Response of Nkedi Zebu and Ankole cattle to tick infestation and natural tick-borne, helminth and trypanosome infections in Uganda

A cross-sectional study was conducted in Soroti district of Uganda to establish important traits of Nkedi Zebu and Ankole cattle regarding their production performance responses to natural infections of trypanosomes, gastrointestinal nematodes, Theileria parva, Babesia bigemina, Anaplasma marginale and tick infestations. Over four visits between October 2006 to August 2007, tick counts were performed and blood, faecal samples and sera were collected from the Nkedi Zebu (295) and Ankole (165) cattle from 86 herds in six locations per visit. Low parasitological prevalence of trypanosome infection (<6%) and high prevalence of gastrointestinal nematode infections (>30%) with low faecal egg counts (110-300 eggs per gramme (EPG)) were observed in the Nkedi Zebu and Ankole cattle. Both breeds had high, moderate and low mean counts of Rhipicephalus appendiculatus (18.0-24.0), Rhipecephalus (Boophilus) decoloratus (3.6-10.3) and Amblyomma variegatum ticks (1.7-4.3), respectively. In addition, both breeds had similar mean packed cell volumes (26.4-31.2) and a similar percentage of animals were anaemic (14.5-36.6%). The Nkedi Zebu cattle further had higher mean optical density (OD) values for antibodies against T. parva (1.093-1.445) and A. marginale infections (0.573-0.583), and significantly (P < 0.001) higher mean OD values of antibodies against B. bigemina infections (1.07-2.175) than the Ankole cattle: T. parva (1.030-1.302); A. marginale (0.442-0.603) and B. bigemina infections (0.863-2.154). The Ankole cows produced significantly more (P < 0.001) milk per day (2.68L) than the Nkedi Zebu cows (1.98L), and the Ankole oxen had significantly higher (P < 0.05) draught power output (2.57days/acre) than the Nkedi Zebu oxen (2.93days/acre). Liveweights of calves aged 0-12months of both breeds were comparable, suggesting that the Nkedi Zebu and Ankole cattle under similar disease challenge exhibited similar growth rates. In conclusion, the Nkedi Zebu cattle seem to possess a higher degree of disease resistance against endemic parasitic diseases, while the Ankole cattle seem to possess a moderate degree of disease resistance coupled with a moderate production potential.

Attraction response of adult Rhipicephalus appendiculatus and Rhipicephalus pulchellus (Acari: Ixodidae) ticks to extracts from Calpurnia aurea (Fabaceae)

Experiments were carried out to investigate the response of two tick species Rhipicephalus pulchellus Gerstaker, 1873 and Rhipicephalus appendiculatus Neumann, 1901 to three different extracts (acetone, aqueous and oil) of the dried leaves of Calpurnia aurea (Aiton) Benth in both an inverted glass tube and a dual choice T-olfactometer. The oil extract at 50 and 100 mg/ml attracted 46.7% and 65.9% of R. appendiculatus, respectively, in the inverted glass tube assay, which was comparable to 47.8% of the attraction-aggregation-attachment pheromone (AAAP) used as positive control. At a dose of 100 mg/ml the oil extract attracted 52.4% of R. pulchellus in the T-olfactometer bioassay. The relative attraction of both tick species to plant extract was also tested in semi-field plot experiments using a trap baited with different concentrations of emulsifiable extract of C. aurea. A dose of 100 mg/ml attracted 52.2% of R. pulchellus and 44.4% of R. appendiculatus from a distance of 1 m while 14.4% of R. pulchellus and 12.2% of R. appendiculatus were attracted from 5 m distance at the same dose. Addition of CO 2 to the plant extract-baited-trap at the dose of 100 mg/ml increased the range of attraction of adult R. pulchellus (44.4% from 5 m distance) and up to 33.3% of adult R. appendiculatus tick from a distance of 4 m. The results of this study suggest that extracts from C. aurea can potentially be used as baits in a trap for the control of ticks in the field.