Rhipicephalus

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. Results Raw data from bTEFAP were screened and trimmed based upon quality scores and binned into individual sample collections. Bacteria identified to the species level include Staphylococcus aureus, Staphylococcus chromogenes, Streptococcus dysgalactiae, Staphylococcus sciuri, Serr...

Green nanoparticle synthesis has been achieved using environmentally acceptable plant extract and eco-friendly reducing and capping agents. The present study was based on assessments of the antiparasitic activities to determine the efficacies of synthesized silver nanoparticles (AgNPs) using aqueous leaf extract of Mimosa pudica Gaertn (Mimosaceae) against the larvae of malaria vector, Anopheles subpictus Grassi, filariasis vector Culex quinquefasciatus Say (Diptera: Culicidae), and Rhipicephalus (Boophilus) microplus Canestrini (Acari: Ixodidae). Parasite larvae were exposed to varying concentrations of aqueous extract of M. pudica and synthesized AgNPs for 24 h. AgNPs were rapidly synthesized using the leaf extract of M. pudica and the formation of nanoparticles was observed within 6 h. The results recorded from UV–vis spectrum, Fourier transform infrared, X-ray diffraction, scanning electron microscopy, and transmission electron microscopy support the biosynthesis and characterization of AgNPs. The maximum efficacy was observed in synthesized AgNPs against the larvae of A. subpictus, C. quinquefasciatus, and R. microplus (LC50 = 13.90, 11.73, and 8.98 mg/L, r 2 = 0.411, 0.286, and 0.479), respectively. This is the first report on antiparasitic activity of the plant extract and synthesized AgNPs.

Rhipicephalus sanguineus, commonly known as “brown dog tick” or “kennel tick”, is a three-host tick that parasitizes mainly dogs and occasionally humans. This arthropod is worldwide distributed and is a vector of some zoonosis, as Rickettsia conorii (pathogen of Mediterranean spotted fever).
In Portugal, R. turanicus present high morphological resemblances to R. sanguineus, and as both populations are sympatric and very genetically similar, are easily misidentified. However, through a careful morphological analysis, it is possible to distinguish them, especially by the use of electronic microscopy.
Although some prior molecular and morphological studies have already been performed on these Portuguese populations, the results differ, which means there is still much speculation and disagreement around this taxonomic classification. 
In this context, this study was conducted using morphological (stereoscopic microscopy), ultramorfological (scanning electron microscopy - SEM) and molecular analysis (COI was the molecular marker of choice) in order to clarify the taxonomic status of these populations. For that purpose, three R. sanguineus s.l. populations collected from the districts of Óbidos, Santarém and Caldas da Rainha were used.
The species identification was performed based on morphology characteristics, using standard taxonomic keys (Dias, 1994; Papadopoulos et al., 1992; Walker et al., 2003). Ticks that could not be morphological identified as R. sanguineus or R. turanicus were defined as intermediate species. Some R. pusillus specimens were included in the analysis as an out-group.
The results obtained with the mentioned methods (except for the molecular analysis), allowed to conclude that there is a great morphological diversity within R. sanguineus s.l. Portuguese populations, and that the presence of R. turanicus in Portugal is grounded. Thus, it is suggested a new taxonomic classification based in observed variability within the group. It also suggested the future use of the statistical methodology employed as a technique for morphological variability evaluation within a representative population.

The present study was based on assessments of the anti-parasitic activities to determine the efficacies of synthesized zinc oxide nanoparticles (ZnO NPs) prepared by wet chemical method using zinc nitrate and sodium hydroxide as precursors and soluble starch as stabilizing agent against the larvae of cattle tick Rhipicephalus (Boophilus) microplus, Canestrini (Acari: Ixodidae); head louse Pediculus humanus capitis, De Geer (Phthiraptera: Pediculidae); larvae of malaria vector, Anopheles subpictus, Grassi; and filariasis vector, Culex quinquefasciatus, Say (Diptera: Culicidae). R. microplus larvae were exposed to filter paper envelopes impregnated with different ZnO NP concentrations. Direct contact method was conducted to determine the potential of pediculocidal activity. Parasite larvae were exposed to varying concentrations of synthesized ZnO NPs for 24 h. The results suggested that the mortality effects of synthesized ZnO NPs were 43% at 1 h, 64% at 3 h, 78% at 6 h, and 100% after 12 h against R. microplus activity. In pediculocidal activity, the results showed that the optimal times for measuring mortality effects of synthesized ZnO NPs were 38% at 10 min, 71% at 30 min, 83% at 1 h, and 100% after 6 h against P. humanus capitis. One hundred percent lice mortality was observed at 10 mg/L treated for 6 h. The mortality was confirmed after 24 h of observation period. The larval mortality effects of synthesized ZnO NPs were 37%, 72%, 100% and 43%, 78% and 100% at 6, 12, and 24 h against A. subpictus and C. quinquefasciatus, respectively. It is apparent that the small size and corresponding large specific surface area of small nanometer-scale ZnO particles impose several effects that govern its parasitic action, which are size dependent. ZnO NPs were synthesized by wet chemical process, and it was characterized with the UV showing peak at 361 nm. X-ray diffraction (XRD) spectra clearly shows that the diffraction peaks in the pattern indexed as the zinc oxide with lattice constants a = 3.249 and c = 5.206 Å. The FTIR spectrum showed the range of 400–4,000 cm−1. The band at 899.56 cm−1; 1,151.87 cm−1; 1,396 cm−1; and these bands showed the complete composition of ZnO NPs. SEM micrograph showed 60–120-nm size and aggregates of spherical shape nanoparticles. EDX showed the complete chemical composition of the synthesized nanoparticles of zinc oxide. The maximum efficacy was observed in zinc oxide against the R. microplus, P. humanus capitis, and the larvae of A. subpictus, C. quinquefasciatus with LC50 values of 29.14, 11.80, 11.14, and 12.39 mg/L; r 2 = 0.805, 0.876, 0.894, and 0.904, respectively. The synthesized ZnO NPs showed the LC50 and r 2 values against the R. microplus (13.41 mg/L; 0.982), P. humanus capitis (11.80 mg/L; 0.966), and the larvae of A. subpictus (3.19; 0.945 mg/L), against C. quinquefasciatus (4.87 mg/L; 0.970), respectively. The control (distilled water) showed nil mortality in the concurrent assay. This is the first report on anti-parasitic activity of the synthesized ZnO NPs.

The formulations of acaripathogenic fungi to control ticks have been widely studied. The present study evaluated the efficacy of oil-based formulations of Metarhizium anisopliae sensu lato (s.l.), isolate Ma 959, and Beauveria bassiana, isolate Bb 986, on different Rhipicephalus microplus stages, comparing the efficacy between aqueous suspensions and 10, 15 and 20% mineral oil formulations. Twelve groups were formed: one aqueous control group; three mineral oil control groups, at 10, 15 or 20%; two aqueous fungal suspensions of M. anisopliae s.l. or B. bassiana; and three formulations of M. anisopliae (s.l.) or B. bassiana containing 10, 15, and 20% mineral oil. To prepare aqueous suspensions and oily formulations, fungal isolates were cultivated on rice grains in polypropylene bags. The conidial suspensions and formulations had a concentration of 10(8)conidia/mL. Bioassays were repeated twice. After treatment, the following biological parameters of engorged females were evaluated: ...

Ticks are obligate haematophagous ectoparasites considered the principal vectors of disease among animals. Rhipicephalus microplus and R. annulatus ticks are the most important vectors for Babesia bigemina and B. bovis, two of the most important intraerythrocytic protozoan parasites species in cattle, responsible for babesiosis which together with anaplasmosis account for substantial economic losses in the livestock industry worldwide. Anti-tick vaccines are a proved alternative to traditional tick and tick borne diseases control methods but are still limited primarily due to the lack of effective antigens. Subsequently to the identification of antigens the validation is a laborious work often expensive. Tick artificial feeding, is a low cost alternative to test antigens allowing achieving critical data. Herein, R. microplus females were successfully artificially fed using capillary tubes. Calreticulin (CRT) protein, which in a previous study has been identified as being involved in...

Acaricide resistance of the cattle tick Rhipicephalus (Boophilus) microplus is widespread in most of the countries where this parasite is present. Bioassays are used to diagnose the level and pattern of resistance in tick populations. In the present study, we describe a detailed protocol of the Larval Tarsal Test (LTT) using simplified equipment and data on the resistance of 17 tick field populations originating from 5 Brazilian states. Nine acaricidal compounds from 5 major classes were tested: organophosphates (OP), synthetic pyrethroids (SP), macrocyclic lactones (ML), phenylpyrazols (PYZ) and amidines. For comparison, four of the tick populations were also tested with the Larval Packet Test (LPT) with one compound per class. The most common resistances were to SP, amitraz and OP, with frequencies of 94%, 88% and 82%, respectively. Resistance to PYZ was also found to be widespread (65%), suggesting a rapid development of fipronil resistance in Brazil. One case of ML resistance an...

The present study evaluated, for the first time, the effect of the commercial formulation Metarril(®) SP Organic of Metarhizium anisopliae plus 10% mineral oil to control Rhipicephalus microplus in a pen study. Three groups were formed with six animals each: the first group was exposed to Metarril(®) plus 10% mineral oil and 1% Tween 80; the second group was exposed to sterile distilled water, mineral oil and Tween 80 (oil control group); and the third group received no treatment (control group). The fungal formulation contained 1 × 10(8)conidiaml(-1). Each animal was sprayed with 3L of formulation. Fallen ticks were counted daily and a sample of 20 engorged females per day was incubated for assessment of biological parameters. Throughout the study period, Metarril(®) oil-based formulation showed an efficacy ranging from 19.20% to 67.39% in comparison with the control group; and from 8.18% to 61.38% in comparison with the oil control group. The average efficacy of Metarril(®) oil-ba...

The toxic effects of the extracts of Allium sativum (Garlic) were evaluated against adults of Hyalomma marginatum rufipes and Rhipicephalus pulchellus using three types (Types A, B and C) of contact toxicity bioassays. A. sativum bulbs were extracted with acetone, ethanol and dichloromethane (DCM) solvents. Among these three solvents, it is the DCM extract of A. sativum that appears to have anti-tick activity. In the Type A contact toxicity bioassay, DCM extracts of A. sativum demonstrated a high acaricidal bioactivity against H. m. rufipes with 100 % of ticks killed in less than an hour, and toxicity persisted to the second day. A weak acaricidal activity of aqueous extracts of A. sativum was observed in the Type B contact toxicity bioassay. In the Type C contact toxicity bioassay, a concentration of 24 % w/v of DCM extracts of garlic in sunflower oil (Helianthus annuus) had killed 100% of H. m. rufipes (LC50= 5.9 % w/v) and R. pulchellus (LC50 = 10.3 % w/v) by 24 hours post-treatm...

Acaricide resistance has become widespread in countries where cattle ticks, Rhipicephalus (Boophilus) microplus, are a problem. Resistance arises through genetic changes in a cattle tick population that causes modifications to the target site, increased metabolism or sequestration of the acaricide, or reduced ability of the acaricide to penetrate through the outer protective layers of the tick’s body. We review the molecular and biochemical mechanisms of acaricide resistance that have been shown to be functional in R. (B.) microplus. From a mechanistic point of view, resistance to pyrethroids has been characterized to a greater degree than any other acaricide class. Although a great deal of research has gone into discovery of the mechanisms that cause organophosphate resistance, very little is defined at the molecular level and organophosphate resistance seems to be maintained through a complex and multifactorial process. The resistance mechanisms for other acaricides are less well ...

Background and Aim: Tick-borne pathogens (TBPs) are of great concern having the potential to threaten canine health. Dogs infected with Ehrlichia canis, Anaplasma platys, Babesia canis, and Hepatozoon canis are commonly found in Thailand; Rhipicephalus sanguineus tick is the most common vector of diseases. This study aimed to determine the prevalence of common TBPs in dogs and their ticks in Thailand using polymerase chain reaction (PCR) and DNA sequencing methods.

Materials and Methods: Forty-four blood samples were positively diagnosed with TBPs infection by microscopy. Samples were from animal hospitals in Maha Sarakham, Amnat Charoen, Nakhon Ratchasima, and Bangkok, Thailand, during January-June 2020. Five to six ticks were also taken from infected dogs, and then, both blood and tick were analyzed using PCR and DNA sequencing.

Results: PCR results showed that R. sanguineus was the only tick species detected in this study. The appearance of single infection with E. canis was the most common infection found in dogs and ticks (64% and 82%, respectively). Correlation of pathogen infection in hosts and their vector was performed by similarity detection of pathogens between blood and tick samples based on PCR analysis in 29 samples (66%) but there was no significant differentiation.

Conclusion: E. canis appears as the most common canine tick-borne pathogen in Thailand, which was detected in both healthy and sick dogs as well as in R. sanguineus. The findings show the relationships among host dogs, pathogens, and ticks. Veterinarians should be proactive in educating pet owners about the risks associated with ticks and their important pathogens and plan effective control strategies.

Background The cattle tick, Rhipicephalus (Boophilus) microplus, economically impact cattle industry in tropical and subtropical regions of the world. The morphological and genetic differences among R. microplus strains have been documented in the literature, suggesting that biogeographical and ecological separation may have resulted in boophilid ticks from America/Africa and those from Australia being different species. To test the hypothesis of the presence of different boophilid species, herein we performed a series of experiments to characterize the reproductive performance of crosses between R. microplus from Australia, Africa and America and the genetic diversity of strains from Australia, Asia, Africa and America. Results The results showed that the crosses between Australian and Argentinean or Mozambican strains of boophilid ticks are infertile while crosses between Argentinean and Mozambican strains are fertile. These results showed that tick strains from Africa (Mozambique) and America (Argentina) are the same species, while ticks from Australia may actually represent a separate species. The genetic analysis of mitochondrial 12S and 16S rDNA and microsatellite loci were not conclusive when taken separately, but provided evidence that Australian tick strains were genetically different from Asian, African and American strains. Conclusion The results reported herein support the hypothesis that at least two different species share the name R. microplus. These species could be redefined as R. microplus (Canestrini, 1887) (for American and African strains) and probably the old R. australis Fuller, 1899 (for Australian strains), which needs to be redescribed. However, experiments with a larger number of tick strains from different geographic locations are needed to corroborate these results.