Displacement of Rhipicephalus decoloratus by Rhipicephalus microplus (Acari: Ixodidae) in the Eastern Cape Province, South Africa

The objective of the study was to establish to what extent the native tick species Rhipicephalus decoloratus has been displaced by the invasive introduced tick, Rhipicephalus microplus at two communally grazed areas in the Eastern Cape Province, South Africa. To this end ticks were collected monthly from five cattle over a period of 2 years and from 10 drag-samples of the vegetation over a period of 1 year at each locality. Whereas 10 years previously only R. decoloratus and no R. microplus had been recorded in the vicinity of the two sites, R. microplus now comprised the bulk of collections at both. Furthermore, significantly more R. microplus were collected from cattle at both localities during the 2nd year of the survey than during the 1st. In addition to 83 instances of intraspecific coupling, there were 17 instances of R. microplus males coupled with R. decoloratus females. Collections made from cattle and goats on two farms close to the study sites revealed that R. microplus was present on both host species and that it significantly outnumbered R. decoloratus on one of the farms. Rhipicephalus decoloratus and R. microplus larvae as well as larvae exhibiting characteristics of both species were collected from the vegetation.     

Geographic distribution of the invasive cattle tick Rhipicephalus microplus, a country-wide survey in Benin

The cattle tick Rhipicephalus microplus is currently invading the West African region, and little information is available on the spread of this exotic tick in this region. We set out a country-wide field survey to determine its current distribution in Benin. Ticks were collected on cattle from 106 farms selected by random sampling covering all regions of the country. Rhipicephalus annulatus was found on 70?% of all farms, R. decoloratus on 42?%, R. geigyi on 58?%, and R. microplus on 49?%. There is a clear geographic separation between the indigenous Rhipicephalus species and R. microplus. Rhipicephalus annulatus occurs mainly in the northern departments, but it was also observed in lower numbers in locations in the south. The presence of R. decoloratus is limited to the northern region, and in most locations, this tick makes up a small proportion of the collected ticks. The tick R. geigyi tends to be dominant, but occurs only in the four northern departments. The observations concerning R. microplus are entirely different, this species occurs in the southern and central region. The results of this survey confirm the invasive character and displacement properties of R. microplus, since in less than a decade it has colonized more than half of the country and has displaced indigenous ticks of the same genus in many of the sampled locations.     

<Emphasis Type="Italic">Rhipicephalus microplus</Emphasis> (Acarina,Ixodidae) in Angola: evidence of its establishment and expansion

Livestock constitutes a fundamental asset for the livelihood of rural communities in Angola. Rhipicephalus microplus (Acarina, Ixodidae) is considered to be the most important external parasite impacting the cattle industry in the world. The present work used a cross-sectional study on tick species infesting cattle in smallholder and commercial farms, to investigate the presence of R. microplus in Angola. This species was found to be recently established in certain areas of Cuanza-Sul and Benguela provinces. Furthermore, taking the current understanding of the pattern of dispersal of R. microplus into account, it is probable that this species is already present in other areas of the country. Animal movement without the necessary control measures facilitates the dispersal of this tick species. Consequently, R. microplus is causing rapid displacement of the native tick species, Rhipicephalus decoloratus. The presence of R. microplus in Angola constitutes a serious problem and poses a potential threat to the livestock sector as well as being an important risk factor for the sustainability of the livelihood strategies developed by smallholders.     

Red deer (Cervus elaphus) as a host for the cattle tick Rhipicephalus microplus (Acari: Ixodidae) in Yucatan, Mexico

Rhipicephalus microplus is the most economically important cattle tick in the Mexican tropics. Wild ungulate species, including red deer (Cervus elaphus), are gaining popularity in diversified livestock ranching operations in Mexico. However, there is no information available on the susceptibility of red deer to infestation with the cattle tick, R. microplus, under hot, subhumid tropical conditions in Mexico. Biological data on R. microplus as an ectoparasite of cattle and red deer in a farm in the Mexican tropics are presented here. Ticks collected from red deer were identified as R. microplus (97 %) and Amblyomma cajennense (3 %), and tick species infesting cattle included R. microplus (95 %) and A. cajennense (5 %). Standard counts of R. microplus engorged females on red deer were 11 times higher than on cattle (428 ± 43 vs. 40 ± 18; p < 0.001). The reproductive efficiency index and larval hatching of R. microplus collected from cattle and red deer were similar (p > 0.05). Hemolymph samples of R. microplus collected from cattle were positive for Babesia spp. (10 %, 2/50) and all the samples from ticks infesting red deer were negative. Seventeen and ten percent of the blood samples from cattle and red deer were positive for Anaplasma marginale, respectively. The role of red deer as a host of R. microplus in Yucatan, Mexico and the importance of this host-parasite relationship relative to the epidemiology of R. microplus-borne diseases are discussed.     

New foci of <Emphasis Type="Italic">Rhipicephalus microplus</Emphasis> in West Africa

The invasive character of Rhipicephalus microplus was observed in Benin, the second West-African country from which this ticks species has been collected after the initial confirmed record in Ivory Coast in 2007. A cross-sectional study was carried out in the Department of Mono to examine the presence of the tick R. microplus. The survey covered 9 herds (villages) in an agro-ecological zone inhabited by agro-pastoralists, including the State Farm of Kpinnou that imported Girolando cattle from Brazil. Almost 800 ticks were sampled from 36 cattle, on average four cattle per village. The morphological identification revealed ticks of two different genera: Rhipicephalus and Amblyomma. Rhipicephalus microplus was the only representative of the species previously known as Boophilus or blue ticks. Its taxonomic identity was confirmed molecularly by PCR–RFLP. A comparison was made with the situation of R. microplus in Brazil.     

Control of tick populations by spraying <Emphasis Type="Italic">Metarhizium anisopliae</Emphasis> conidia on cattle under field conditions

Conidia of the entomopathogenic fungus, Metarhizium anisopliae, in oil/water formulation (1 × 10⁸ conidia/ml) were sprayed at 3 weekly intervals on Rhipicephalus evertsi evertsi and Rhipicephalus (Boophilus) decoloratus ticks while feeding on Afrikana bulls grazing in paddocks for a period of 1 year. The fungus reduced the on-host tick populations by 83% 3 month after commencement of the experiment. The formulation by itself had only minimal effect on the tick population. Tick populations and fungal efficacy were highest at the peaks of rainfall and relative humidity or soon thereafter. Fed and unfed adult R. e. evertsi and R. (B.) decoloratus collected at the end of the experiment from the fungus-sprayed and from the control cattle and incubated in the laboratory exhibited a mortality of 93% in oil formulated conidia and 14% in oil control. The corresponding mortality in R. (B.) decoloratus was 100% in fungus and 11% in oil control. Ticks on the fungus-sprayed groups had significantly higher mortality (P < 0.05) than on the control groups. Furthermore, no significant difference (P < 0.05) was observed in fungus-induced mortality between the two tick species. Mortalities induced by Triton X-100 (0.05%), sunflower oil (20%) and water alone were low, suggesting that they were non-toxic to ticks at the concentrations used and no significant difference was observed among them. No physical or behavioral abnormalities were observed in the fungus-sprayed cattle at any time during the course of the experiment. All groups of cattle gained weights during the experimental period.     

Survival and behaviour of unfed stages of the ticksRhipicephalus appendiculatus,Boophilus decoloratus andB. microplus under field conditions in Zimbabwe

The survival and behaviour of the unfed stages ofRhipicephalus appendiculatus, Boophilus decoloratus andB. microplus in gauze columns were observed in long and short grass in the highveld of Zimbabwe. Ticks were exposed in the cool, hot and rainy seasons of 1980 and 1981. All species and stages survived longer in long grass than in short grass. Larvae from engorged female ticks released in the cool season hatched much later than incubator-reared controls. They were consequently not present during the cold weather and survived longer than larvae subjected to the low temperatures, in which the shortest survival-times were recorded. The survival of nymphs was insensitive to season. The longest survival-times were recorded in adults. Median survival-times of incubator-reared adults ranged from 165 to 375 days in short grass and from 333 to 493 days in long grass. These times were usually longer than those for adults which moulted in the field. Larvae of the three species and nymphs ofR. appendiculatus were active soon after hatching or moulting, irrespective of the season. In contrast, adults ofR. appendiculatus showed different patterns of activity in different seasons. Adults first appeared at the base of the columns in October/November and then gradually ascended to reach a maximum height in December/January. They remained high up in the columns until May/June when the weather became increasingly cold and dry. Larvae ofB. decoloratus climbed higher up in the columns in the long grass than did the larvae of the other two species.Larvae and nymphs ofR. appendiculatus and larvae ofB. microplus migrated up and down the columns daily, but larvae ofB. decoloratus and adults ofR. appendiculatus did not migrate.     

Hard tick species (Acari: Ixodidae) and infestation in two livestock agroecosystems from Antioquia,Colombia

Tick infestation affects about 80% of livestock globally while transmitting various pathogens causing high economic losses. This study aimed to determine the degree of tick infestation in two regions, North and Middle Magdalena in Antioquia, Colombia, to identify the ixodid tick species found and the associated risk factors. A cross-sectional study was carried out in 48 farms distributed in six municipalities of Antioquia. Two paddocks and eight bovines per farm were evaluated to estimate tick infestation (adults, nymphs, and larvae). Tick species were identified through a morphological and molecular analysis based on partial sequences of data obtained from DNA molecular markers, two mitochondrial (16S rRNA and COI), and one genomic DNA gene (18S rRNA). A multivariate Poisson regression model was applied to estimate the associated risk factors with ticks in cattle. Rhipicephalus microplus, Amblyomma patinoi and Dermacentor nitens were present in the livestock agroecosystems in the Middle Magdalena region; the highest incidence of tick infestation in cows and paddocks was reported in the municipality of Puerto Triunfo. The livestock agroecosystems in Middle Magdalena were characterized by a higher presence of adult R. microplus in cattle. Larval infestation of R. microplus, followed by D. nitens, was also found in paddocks. The multivariate analysis showed that the origin of cattle was the main risk factor associated with the presence of ticks (i.e., when cattle came from outside the farm). Cattle movement between farms in Middle Magdalena can contribute to the spread of ticks in this region.

     

Electrospun micro- and nanofiber tubes for functional nervous regeneration in sciatic nerve transections

Background

Rhipicephalus (Boophilus) spp. ticks economically impact on cattle production in Africa and other tropical and subtropical regions of the world. Tick vaccines constitute a cost-effective and environmentally friendly alternative to tick control. The R. microplus Bm86 protective antigen has been produced by recombinant DNA technology and shown to protect cattle against tick infestations.

Results

In this study, the genes for Bm86 (R. microplus), Ba86 (R. annulatus) and Bd86 (R. decoloratus) were cloned and characterized from African or Asian tick strains and the recombinant proteins were secreted and purified from P. pastoris. The secretion of recombinant Bm86 ortholog proteins in P. pastoris allowed for a simple purification process rendering a final product with high recovery (35–42%) and purity (80–85%) and likely to result in a more reproducible conformation closely resembling the native protein. Rabbit immunization experiments with recombinant proteins showed immune cross-reactivity between Bm86 ortholog proteins.

Conclusion

These experiments support the development and testing of vaccines containing recombinant Bm86, Ba86 and Bd86 secreted in P. pastoris for the control of tick infestations in Africa.     

Ticks (Acari: Ixodidae) associated with domestic dogs in Franca region,São Paulo,Brazil

Ticks on 140 domestic dogs from both urban and rural areas of Franca region in São Paulo state were identified with 102 dogs from urban areas and 38 from rural areas. Of urban dogs, 27.5% were infested exclusively by Rhipicephalus sanguineus ticks. Of the rural dogs, 36.8% were infested with the following tick species: R. sanguineus, Boophilus microplus, Amblyomma ovale and A. cajennense. Mixed infestations included a dog hosting A. cajennense and A. ovale and another with B. microplus and R. sanguineus. The most intense infestations were detected on urban dogs. Hemolymph tests of these ticks performed to detect rickettsial or Borrelia bacteria yielded negative results.     

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.     

Assessment of bacterial diversity in the cattle tick Rhipicephalus (Boophilus) microplus through tag-encoded pyrosequencing

Background  

Ticks are regarded as the most relevant vectors of disease-causing pathogens in domestic and wild animals. The cattle tick, Rhipicephalus (Boophilus) microplus, hinders livestock production in tropical and subtropical parts of the world where it is endemic. Tick microbiomes remain largely unexplored. The objective of this study was to explore the R. microplus microbiome by applying the bacterial 16S tag-encoded FLX-titanium amplicon pyrosequencing (bTEFAP) technique to characterize its bacterial diversity. Pyrosequencing was performed on adult males and females, eggs, and gut and ovary tissues from adult females derived from samples of R. microplus collected during outbreaks in southern Texas.     

Ticks and Tick-Borne Diseases of Livestock Belonging to Resource-poor Farmers in the Eastern Free State of South Africa

The paper provides a summary of three studies conducted in the eastern Free State of South Africa between 1998 and 2000. In a questionnaire-based study approximately 21% of interviewed resource-poor farmers (n = 150) indicated that they experienced problems with ticks and tick-borne diseases. About 56% of farmers indicated that tick-related problems were most severe in summer, while 32% indicated that the most problems were encountered in winter. About 12% indicated that the tick problems were experienced throughout the year. Farmers also indicated that the highest tick burdens were experienced between spring and early winter. The principal ticks infesting cattle (n = 30) were found to be Boophilus decoloratus (53.1%), Rhipicephalus evertsi evertsi (44.7%), Rhipicephalus follis (1.0%), Rhipicephalus gertrudae (0.7%) and Rhipicephalus warburtoni (0.4%). On small stock (n = 188), R. evertsi evertsi (68%) and B. decoloratus (32%) were recorded as the main ticks in the study area. A sero-epidemiological survey of cattle (n = 386) showed that 94% of cattle were sero-positive for Babesia bigemina by IFAT, while 87% were sero-positive for Anaplasma by indirect ELISA. All the animals were sero-negative for Babesia bovis and this is probably because the tick vector, Boophilus microplus, is not present in the study area. All sheep and goats were sero-positive for Theileria species by IFAT while 85% of sheep and 100% of goats tested positive for Anaplasma species by competition inhibition ELISA. The high incidence of positive serological results for B. bigemina and Anaplasma in cattle, and Theileria and Anaplasma in sheep and goats and the absence of clinical cases would indicate that this area is endemically stable for these diseases. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cross immunity with Haemaphysalis longicornis glutathione S-transferase reduces an experimental Rhipicephalus (Boophilus) microplus infestation

Recombinant Glutathione S-transferase of Haemaphysalis longicornis (rGST-Hl) was expressed in Escherichia coli, purified by affinity chromatography and used in the immunization of cattle. Western blot analysis showed positive antibody response in cattle immunized with rGST-Hl. The tests also showed that immunized bovine sera recognize native Rhipicephalus microplus proteins in different tissue extracts. Furthermore, the vaccine potential of rGST-Hl was investigated against infestation of Hereford cattle by R. microplus. Vaccination of cattle with rGST-Hl conferred partial cross-protection immunity against R. microplus. Considering the effect on number of engorged ticks, egg laying capacity and egg fertility, the overall efficacy of vaccination was of 57%, as compared with control group.     

Genetic diversity and phylogenetic analyses of ixodid ticks infesting cattle in northeast Thailand: the discovery of Rhipicephalus microplus clade C and the rarely detected R. haemaphysaloides

In total, 160 ticks infesting cattle in the northeast region of Thailand were collected and used for molecular investigation. Three tick species—Rhipicephalus microplus Canestrini, Rhipicephalus haemaphysaloides Supino and Haemaphysalis bispinosa Neumann—were identified based on morphology and DNA sequences of mitochondrial cytochrome c oxidase subunit 1 (CO1) and 16S ribosomal RNA (16S rRNA). In total, 26 and seven unique haplotypes of the CO1 and 16S rRNA genes, respectively, were recovered. Phylogenetic analysis using the CO1 sequence revealed that the R. microplus from northeastern Thailand were grouped into the previously described clades A and C, whereas the 16S rRNA phylogenetic tree assigned all isolates of R. microplus from Northeast Thailand into the previously described clade B. Clade C of the CO1 phylogenetic tree is a new genetic assemblage recently discovered from India and Malaysia, which has now been detected in our study. The haplotype network also demonstrated that R. microplus is divided into two haplogroups corresponding to the assemblage of the CO1 phylogenetic tree. Our findings strongly support the previous genetic assemblage classification and evidence that R. microplus from Northeast Thailand is a species complex comprising at least two genetic assemblages, i.e., clades A and C. However, further investigation is needed and should involve more comprehensive genetic and morphological analyses and cover a larger part of their distributional range throughout Southeast Asia.

     

Shifts in the distributional ranges of <Emphasis Type="Italic">Boophilus</Emphasis> ticks in Tanzania: evidence that a parapatric boundary between <Emphasis Type="Italic">Boophilus microplus</Emphasis> and <Emphasis Type="Italic">B. decoloratus</Emphasis> follows climate gradients

Based on a national tick survey conducted in Tanzania between 1998 and 2001, predictive maps of tick distribution for B. microplus and B. decoloratus were computed and compared with historical maps that originated about 40 years ago. The new data collected showed that except for extremely cold and dry areas B. microplus has extended its distribution range and is now present in all the northern regions of Tanzania, and that high suitability is currently recorded for most of the previously non-occupied areas. In contrast, B. decoloratus, once abundant in Tanzania, has largely retreated to highlands in north and central Tanzania and several other high-altitude refuges. Geostatistical analysis revealed a dissimilar character of distribution for the two species at a local and regional scale, as well as an antagonistic relationship between them in a spatial context. The Tanzania data suggest that an equilibrium phase with a stable parapatric boundary that follows a climate gradient has been achieved. The zone of overlap in the distribution ranges of B. microplus and B. decoloratus is characterized by a decreased infestation rate as well as mutual exclusion on infested cattle. This finding matches studies on parapatry with hybrid zones, where the latter zones that separate species tend to rest in density troughs. Analysis of the Tanzania data shows that the critical factor for the advance of B. microplus and the retreat of B. decoloratus is associated with the 58 mm isohyet and the 22–23°C isotherm and indicates a well developed higher-temperature tolerance for B. microplus. It can be anticipated that climate changes may enhance the spread of B. microplus and consequently Babesia bovis into new areas of the African continent.     

Thrombin induces Egr-1 expression in fibroblasts involving elevation of the intracellular Ca2+ concentration, phosphorylation of ERK and activation of ternary complex factor

Background

The Arthropods are a diverse group of organisms including Chelicerata (ticks, mites, spiders), Crustacea (crabs, shrimps), and Insecta (flies, mosquitoes, beetles, silkworm). The cattle tick, Rhipicephalus (Boophilus) microplus, is an economically significant ectoparasite of cattle affecting cattle industries world wide. With the availability of sequence reads from the first Chelicerate genome project (the Ixodes scapularis tick) and extensive R. microplus ESTs, we investigated evidence for putative RNAi proteins and studied RNA interference in tick cell cultures and adult female ticks targeting Drosophila homologues with known cell viability phenotype.

Results

We screened 13,643 R. microplus ESTs and I. scapularis genome reads to identify RNAi related proteins in ticks. Our analysis identified 31 RNAi proteins including a putative tick Dicer, RISC associated (Ago-2 and FMRp), RNA dependent RNA polymerase (EGO-1) and 23 homologues implicated in dsRNA uptake and processing. We selected 10 R. microplus ESTs with >80% similarity to D. melanogaster proteins associated with cell viability for RNAi functional screens in both BME26 R. microplus embryonic cells and female ticks in vivo. Only genes associated with proteasomes had an effect on cell viability in vitro. In vivo RNAi showed that 9 genes had significant effects either causing lethality or impairing egg laying.

Conclusion

We have identified key RNAi-related proteins in ticks and along with our loss-of-function studies support a functional RNAi pathway in R. microplus. Our preliminary studies indicate that tick RNAi pathways may differ from that of other Arthropods such as insects.     

Babesia bigemina,Babesia argentina, and Anaplasma marginale: Coinfectious immunity in bovines

Forty-eight intact and eight splenectomized cattle were used to evaluate different systems of coinfectious immunization against Babesia bigemina, Babesia argentina, and Anaplasma marginale. Coinfectious immunity was induced by two methods: (1) blood of cattle acutely infected with B. bigemina, B. argentina and A. marginale was used as the source of inoculum and the post vaccination reactions were chemotherapeutically controlled with Imidocarb, Ganaseg, Gloxazone, and Liquamycin, and (2) by artificially inducing babesiosis with the blood of carrier cattle with chronic infections of B. bigemina and B. argentina without chemotherapy. The degree of resistance was determined by bloodborne and tick-borne challenges. Ticks were collected from cattle and identified as Boophilus microplus and Dermacentor nitens. Vaccinated cattle demonstrated a high degree of resistance to babesiosis and anaplasmosis; however, cattle without coinfectious immunity were treated chemotherapeutically to prevent death losses.     

Acaricidal activity of Metarhizium anisopliae isolated from paddocks in the Mexican tropics against two populations of the cattle tick Rhipicephalus microplus

The acaricidal effects of 55 strains of Metarhizium anisopliae (Metschnikoff) Sorokin, 1883 (Hypocreales: Clavicipitaceae) isolated from paddocks of cattle farms were evaluated in two Rhipicephalus microplus (Canestrini 1887) (Ixodida: Ixodidae) populations, of which one was multi‐resistant and one was susceptible to chemical acaricides. Percentage mortality and reproductive efficiency indices in R. microplus were evaluated by adult immersion tests at a dose of 1 × 10⁸ conidia/mL for each fungal strain. Some strains were selected to calculate lethal concentrations to kill 50% (LC₅₀) and 99% (LC₉₉) of engorged ticks. Strains MaV22, MaV26 and MaV55 induced 100% mortality in R. microplus on day 14. Strains MaV05, MaV09 and MaV22 caused mortality of >90% from day 12 onward in both tick populations. The most effective acaricidal fungal strain, MaV55, inhibited egg laying by 54.86 and 55.86% in acaricide‐resistant and ‐susceptible R. microplus populations, respectively. None of the fungal strains had statistically significant effects on larval hatching. In conclusion, nine strains of M. anisopliae demonstrated high acaricidal effects against R. microplus and reduced its egg laying. No differences in acaricidal effects were observed between the two populations of ticks tested.