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Sample records for garrapata boophilus microplus

  1. Novel approaches for control of Rhipicephalus (Boophilus) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We review recent progress for control of Rhipicephalus (Boophilus) microplus and R. (B.) annulatus. Outbreak infestations in the U.S. have reached alarming levels, due to increased populations of deer and other ungulates serving as alternative hosts. GIS mapping of infestations and deer habitat aids...

  2. Efficacy of Azadirachta indica extracts against Boophilus microplus.

    PubMed

    Srivastava, Rahul; Ghosh, S; Mandal, D B; Azhahianambi, P; Singhal, P S; Pandey, N N; Swarup, D

    2008-12-01

    Extracts were prepared from leaf, bark, and seed of Azadirachta indica, leaf and seed of Prunus persica, bark of Mangifera indica, and leaf of Psidium guajava and were evaluated against Boophilus microplus. Of the eight extracts screened, the extracts prepared from the A. indica seed showed very high level of efficacy (80%) after 5 h of treatment. Besides the immediate effect on adult ticks, the egg-laying properties of the survived ticks was also assessed, and a significant reduction (P < 0.01) in the reproductive index of ticks fed on animals treated with A. indica seed extracts was noted in comparison to control. The efficacy of the neem seed extracts was compared with the commonly used synthetic pyrethroids, and comparable efficacy against B. microplus fed on animals treated with neem seed extracts and acaricide treated was noted. The possibility of using the extracts in IPM format for the management of ticks is discussed.

  3. Immunoprotective potential of a Rhipicephalus (Boophilus) microplus metalloprotease.

    PubMed

    Ali, Abid; Parizi, Luís Fernando; Guizzo, Melina Garcia; Tirloni, Lucas; Seixas, Adriana; Vaz, Itabajara da Silva; Termignoni, Carlos

    2015-01-15

    Ticks have serious impacts on animal and human health, causing significant economic losses in cattle breeding. Besides damage due to the hematophagous behavior, they transmit several pathogens. Low cost and environmental safety have made vaccines a promising alternative control method against tick infestation. Metalloproteases (MPs) have been shown to be essential for diverse biological functions in hematophagous organisms, inhibiting blood clotting, degrading extracellular matrix proteins, and inhibiting host tissue repair via anti-angiogenic activity. In this study, we analyzed the immunoprotective potential of a recombinant MP against Rhipicephalus (Boophilus) microplus infestation. First, a cDNA encoding R. microplus amino acids sequence with highly conserved regions of the metzincin (reprolysin) group of MP was identified (BrRm-MP4). After expression and purification, recombinant BrRm-MP4 was used as a vaccinal antigen against R. microplus infestation in cattle (Bos taurus taurus). All vaccinated bovines developed immune response to the antigen, resulting in increased antibody level throughout the immunization protocol. Immunization with rBrRm-MP4 reduced tick feeding success, decreasing the number of engorged females and their reproduction potential, representing a 60% overall protection. These results show that rBrRm-MP4 provides protection against tick infestation, placing it is a potential candidate for an anti-tick vaccine.

  4. Ixodicide activity of Eysenhardtia polystachya against Rhipicephalus (Boophilus) microplus.

    PubMed

    Gutierrez, L; Sumano, H; Rivero, F; Alcala-Canto, Y

    2015-04-01

    The objective of this study was to evaluate the acaricidal effect of 2 compounds extracted from Eysenhardtia polystachya in vitro, namely coatlines and matlalines. Each extract was obtained by aqueous extraction in a nitrogen atmosphere. Engorged adult females of an amitraz-resistant strain of Rhipicephalus (Boophilus) microplus were used to produce larvae and to carry out the adult immersion test (AIT), while larvae were assayed using the larval packet test (LPT). After exposure to coatlines or matlalines, the mortality of larvae and engorged females, as well as the reproductive parameters of adult ticks were assessed. Coatlines were not lethal against larvae and did not decrease reproductive parameters. Conversely, matlalines showed a 90 to 100% efficacy against adults and larvae at all concentrations tested and a reduction of 76.41 to 80.64% oviposition and a 48.02 to 54.86% reduction in egg hatchability. Therefore, the acaricidal activity of matlalines was more efficient. Further studies are required to elucidate both the mode of action of matlalines as well the structure-activity relationships responsible for the observed differential efficacy between these 2 related isoflavans against the cattle tick.

  5. Status of Nilgai antelope as a host of cattle fever tick (Boophilus microplus) and Babesiosis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In 2014 an outbreak of cattle fever ticks, Boophilus microplus, was discovered in Cameron County Texas. It was suspected that the ticks had spread from Mexico into Texas carried by Nilgai antelope, a bovid game animal that ranges freely, across the river between Mexico and the US. Between 9 October ...

  6. The complexity of Rhipicephalus (Boophilus) microplus genome characterised through detailed analysis of two BAC clones

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Rhipicephalus (Boophilus) microplus (Rmi) a major cattle ectoparasite and tick borne disease vector, impacts on animal welfare and industry productivity. In arthropod research there is an absence of a complete Chelicerate genome, which includes ticks, mites, spiders, scorpions and crusta...

  7. Microarray analysis of gene expression in acaricide-exposed Rhipcephalus (Boophilus) microplus larvae.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Acaricide-inducible differential gene expression was studied in larvae of Rhipicephalus (Boophilus) microplus using a microarray-based approach. The acaricides used were: coumaphos, permethrin, ivermectin, and amitraz. The microarrays contained over 13,000 probes, having been derived from a previous...

  8. Targeted silencing of the aquaporin 2 gene of rhipicephalus (Boophilus) microplus reduces tick fitness

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ticks are blood-feeding arthropods that can affect human and animal health both directly by up taking blood and indirectly by transmitting pathogens. The cattle tick Rhipicephalus (Boophilus) microplus is one of the most economically important ectoparasites of bovines and it is responsible for the t...

  9. Inheritance of pyrethroid resistance and a sodium channel gene mutation in the cattle tick Boophilus microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A single nucleotide polymorphism (SNP) producing a substitution (Phe'Ile), within the S6 transmembrane segment at domain III within the sodium channel gene sequence, has been associated with pyrethroid resistance in the cattle tick Boophilus microplus. The aim of the present study was to analyze the...

  10. New insights from molecular characterization of the tick Rhipicephalus (Boophilus) microplus in Brazil.

    PubMed

    Csordas, Bárbara Guimarães; Garcia, Marcos Valério; Cunha, Rodrigo Casquero; Giachetto, Poliana Fernanda; Blecha, Isabella Maiumi Zaidan; Andreotti, Renato

    2016-01-01

    The Rhipicephalus (Boophilus) microplus complex currently consists of five taxa, namely R. australis, R. annulatus, R. (B.) microplus clade A sensu, R. microplus clade B sensu, and R. (B.) microplus clade C sensu. Mitochondrial DNA-based methods help taxonomists when they are facing the morpho-taxonomic problem of distinguishing members of the R. (B.) microplus complex. The purpose of this study was to perform molecular characterization of ticks in all five regions of Brazil and infer their phylogenetic relationships. Molecular analysis characterized 10 haplotypes of the COX-1 gene. Molecular network analysis revealed that haplotype H-2 was the most dispersed of the studied populations (n = 11). Haplotype H-3 (n = 2) had the greatest genetic differentiation when compared to other Brazilian populations. A Bayesian phylogenetic tree of the COX-1 gene obtained strong support. In addition, it was observed that the population of R. (B.) microplus haplotype H-3 exhibited diverging branches among the other Brazilian populations in the study. The study concludes that the different regions of Brazil have R. (B.) microplus tick populations with distinct haplotypes.

  11. Rhipicephalus (Boophilus) microplus resistant to acaricides and ivermectin in cattle farms of Mexico.

    PubMed

    Rodríguez-Vivas, Róger Iván; Pérez-Cogollo, Luis Carlos; Rosado-Aguilar, José Alberto; Ojeda-Chi, Melina Maribel; Trinidad-Martinez, Iris; Miller, Robert John; Li, Andrew Yongsheng; de León, Adalberto Pérez; Guerrero, Félix; Klafke, Guilherme

    2014-01-01

    Ticks and the diseases they transmit cause great economic losses to livestock in tropical countries. Non-chemical control alternatives include the use of resistant cattle breeds, biological control and vaccines. However, the most widely used method is the application of different chemical classes of acaricides and macrocyclic lactones. Populations of the cattle tick, Rhipicephalus (Boophilus) microplus, resistant to organophosphates (OP), synthetic pyrethroids (SP), amitraz and fipronil have been reported in Mexico. Macrocyclic lactones are the most sold antiparasitic drug in the Mexican veterinary market. Ivermectin-resistant populations of R. (B.) microplus have been reported in Brazil, Uruguay and especially in Mexico (Veracruz and Yucatan). Although ivermectin resistance levels in R. (B.) microplus from Mexico were generally low in most cases, some field populations of R. (B.) microplus exhibited high levels of ivermectin resistance. The CHPAT population showed a resistance ratio of 10.23 and 79.6 at lethal concentration of 50% and 99%, respectively. Many field populations of R. (B.) microplus are resistant to multiple classes of antiparasitic drugs, including organophosphates (chlorpyrifos, coumaphos and diazinon), pyrethroids (flumethrin, deltamethrin and cypermethrin), amitraz and ivermectin. This paper reports the current status of the resistance of R. (B.) microplus to acaricides, especially ivermectin, in Mexican cattle.

  12. Detection of genomic variability in different populations of the cattle tick Boophilus microplus in southern Brazil.

    PubMed

    Passos, D T; Ferreira, C A; da Silva, S S; Richter, M F; Ozaki, L S

    1999-11-01

    DNA from seven isolates of the cattle tick Boophilus microplus was analyzed by restriction fragment length polymorphism (RFLP). Three different cDNA clones, named P-9, P-25 and CP-12, isolated from a B. microplus cDNA library, were used as DNA probes. DNA sequences of P-9 have high similarity to ribosomal genes, whereas P-25 does not show significant homology with known sequences within databases. CP-12 is a cDNA clone encoding a cysteine endopeptidase gene. A limited degree of polymorphism was detected with P-9 and P-25, while CP-12 showed a different pattern of bands for each tick isolate. These findings suggest the existence of a complex genotypic diversity of the tick B. microplus population in endemic regions.

  13. Presumptive evidence for the role of the white-tailed deer in the epidemiology of Rhipicephalus (Boophilus) annulatus and Rhipicephalus (Boophilus) Microplus (Acari: Ixodidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    From 1907 when the fever tick eradication campaign began until 1933 when the tick eradication methods of dipping cattle in an acaricide and “pasture vacation” failed to eradicate Rhipicephalus (Boophilus) microplus from parts of Florida, the two methods for tick eradication were used with success. ...

  14. Immunological control of ticks through vaccination with Boophilus microplus gut antigens.

    PubMed

    De La Fuente, J; Rodríguez, M; García-García, J C

    2000-01-01

    The control of tick infestations and the transmission of tick-borne diseases remain a challenge for the scientific community. Traditional control methods have been only partially successful. Recently, vaccination with recombinant Boophilus microplus gut antigens has been shown to control tick infestations. Our Bm86-containing vaccine formulation (Gavac) has been effective for the control of artificial infestations of B. annulatus, B. decoloratus, and chemically sensitive and resistant B. microplus strains from Australia, Africa, America, and Iran. Preliminary results with Hyalomma spp. and Rhipicephalus spp. suggest partial cross protection. In field trials, vaccination with Gavac controlled B. microplus and B. annulatus infestations and reduced the transmission of babesiosis, resulting in important savings for the cattle industry. Different degrees of susceptibility to the vaccination with Bm86 and sequence variations in the Bm86 locus have been reported. The Bm95 antigen was isolated from the Argentinean Bm86-resistant B. microplus strain A. A Bm95-based vaccine was used to protect cattle against tick infestations under production conditions with similar results to that obtained with Gavac. The Bm95 antigen from strain A was able to protect against infestations with Bm86-sensitive and Bm86-resistant tick strains, thus suggesting that Bm95 could be a more universal antigen in protecting cattle against infestations by B. microplus strains from different geographical areas. These results clearly demonstrate the advantage and possibilities for the immunological control of ticks.

  15. Natural occurrence of lethal aspergillosis in the cattle tick Rhipicephalus (Boophilus) microplus (Acari:Ixodidae).

    PubMed

    Miranda-Miranda, E; Cossio-Bayugar, R; Martínez-Ibañez, F; Casasanero-Orduña, R; Folch-Mallol, J

    2012-02-01

    The purpose of this study was to describe an unreported entomopathogenic fungus that naturally infects the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae). Engorged female ticks, showed symptoms of fungal infection after controlled tick infestation of cattle. Infected ticks developed a distinctive dark colour, a pale mould grew over the cuticle and the ticks eventually died covered with fungal conidiophores. The responsible fungus was isolated and cultured on mycological medium and submitted to microscopic morphology, biochemical phenotyping and 18S rRNA ribotyping analyses, which identified it as aflatoxin-producing Aspergillus flavus. Spores from the cultured fungus were experimentally sprayed over healthy engorged female ticks, obtaining an 80% prevalence of experimental infection of healthy ticks and their egg masses, the larval progeny after incubation under laboratory conditions was also infected. These results demonstrate that A. flavus is the causative agent of the natural fungal disease of the cattle tick R. microplus described here.

  16. Emergence of oriental theileriosis in cattle and its transmission through Rhipicephalus (Boophilus) microplus in Assam, India

    PubMed Central

    Kakati, Parikshit; Sarmah, Prabhat Chandra; Ray, Debdatta; Bhattacharjee, Kanta; Sharma, Rajeev Kumar; Barkalita, Luit Moni; Sarma, Dipak Kumar; Baishya, Bhaben Chandra; Borah, Pranjal; Stanley, Bobitha

    2015-01-01

    Aim: The aim of the present study was to investigate the presence of Theileria in blood samples of crossbred and indigenous adult cows raised under unorganized small scale farming system in a Babesia and Anaplasma endemic geographical area from Assam, India and to see its transmission through Rhipicephalus (Boophilus) microplus ticks. Materials and Methods: For the present study, 57 clinical cases of cattle suspected to be of hemoparasitic infections were taken into consideration. The parasites were identified based on morphology in giemsa stained blood smear followed by polymerase chain reaction (PCR). Sera samples were tested for T. annulata antibodies in plate and Dot-ELISA. PCR was also conducted in eggs of Rhipicephalus (Boophilus) microplus tick collected from a Theileria orientalis positive animal. Results: PCR amplified 1124, 776, and 160 bp DNA fragments of B. bigemina (64.91%), T. orientalis (21.05%) and A. marginale (14.03%), respectively. This assay further conducted in 12 T. orientalis positive blood samples with primers of Buffeli, Chitose, and Ikeda variants of T. orientalis showed 3 samples positive to Ikeda type and none for Buffeli and Chitose. Babesia bovis and Theileria annulata specific primers also did not amplify any fragment during the PCR assay of the blood samples. Further, all sera samples tested negative to T. annulata antibodies in Plate and Dot-ELISA. PCR conducted in eggs of R (B).microplus tick collected from a T. orientalis positive animal revealed presence of the parasite DNA. Gradual improvement in physical condition leading to complete recovery in 10 out of 12 T. orientalis infected clinical cases treated with buparvaquone(at 2.5mg/kg.b.wt I/M) was the feedback obtained from field veterinarians and the cattle owners. Conclusion: The present investigation represents the first report of occurrence of T. orientalis in cattle of Assam with involvement of pathogenic Ikeda strain in clinical outbreaks and its possible natural

  17. Research and development of anti-tick vaccines for use in Texas and Puerto Rico Rhipicephalus (Boophilus) microplus control programs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This year marks the first time anti-tick vaccination will be used in the United States and Puerto Rico to control, Rhipicephalus (Boophilus) microplus and R. annulatus. The 110-year-old Cattle Fever Tick Eradication Program has eradicated the southern cattle fever tick from the majority of the Unite...

  18. Inhibitor Profile of bis(n)-tacrines and N-methylcarbamates on Acetylcholinesterase from Rhipicephalus (Boophilus) microplus and Phlebotomus papatasi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cattle tick, Rhipicephalus (Boophilus) microplus (Bm), and the sand fly, Phlebotomus papatasi (Pp), are disease vectors to cattle and humans, respectively. The purpose of this study was to characterize the inhibitor profile of acetylcholinesterases from Bm (BmAChE1) and Pp (PpAchE) compared to h...

  19. Genetic factors potentially reducing fitness cost of organophosphate-insensitive acetylcholinesterase(s) in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Acaricidal activity of organophosphate (OP) and carbamate acaricides is believed to result from inhibition of acetylcholinesterase (AChE). Previous studies in Rhipicephalus (Boophilus) microplus demonstrated the presence of three presumptive AChE genes (BmAChEs). Biochemical characterization of re...

  20. Acetylcholinesterases of Rhipicephalus (Boophilus) microplus and Phlebotomus papatasi: Gene identification, expression and biochemical properties of recombinant proteins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rhipicephalus (Boophilus) microplus (Bm) is a vector of bovine babesiosis and anaplasmosis. Tick resistance to organophosphate (OP) acaricide involves acetylcholinesterase (AChE) insensitivity to OP and metabolic detoxification. In vitro expression of Bm genes encoding AChE allowed biochemical chara...

  1. Acetylcholinesterase of Rhipicephalus (Boophilus) microplus and Phlebotomus papatasi: Gene identification, expression, and biochemical properties of recombinant proteins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rhipicephalus (Boophilus) microplus (Bm) ticks are vectors of bovine babesiosis and anaplasmosis. Tick resistance to organophosphate (OP) acaricide involves acetylcholinesterase (AChE) insensitivity to OP and metabolic detoxification. Sequencing and in vitro expression of Bm genes encoding AChE allo...

  2. Serial analysis of gene expression in Rhipcephalus (Boophilus) microplus following organophoosphate treatment of larvae from organophosphate resistant and susceptible strains.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Organophosphate resistant and susceptible tick larvae from laboratory strains of the southern cattle tick, Rhipicephalus (Boophilus) microplus were exposed to low doses of the organophosphate (OP) acaricide, coumaphos. Serial analysis of gene expression (SAGE) was used to analyze differential gene e...

  3. Tests to determine LC50 and discriminating concentrations for fipronil against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) and their standardization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Laboratory test were carried out on larvae and adults of the cattle tick, Rhipicephalus (Boophilus) microplus, to determine fipronil toxicity. Adult immersion test (AIT), larval immersion test (LIT) and larval packet test (LPT) were standardized using susceptible strain (Mozo). Curves dose-response ...

  4. Molecular and functional characterization of the first tick CAP2b (periviscerokinin) receptor from Rhipicephalus (Boophilus) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cDNA of the receptor for CAP2b/periviscerokinin (PVK) neuropeptides, designated Rhimi-CAP2b-R, was cloned from synganglia of tick Rhipicephalus (Boophilus) microplus. This receptor is the ortholog of the insect CAP2b/PVK receptor, as concluded from analyses of the predicted protein sequence, ph...

  5. Transcriptome database derived from the Texas Deutsch outbreak strain population of the cattle tick, Rhipicephalus (Boophilus) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Southern cattle tick, Rhipicephalus (Boophilus) microplus, vectors Babesia bovis and B. bigemina, which are the protozoans causing cattle fever, a disease that is responsible for significant production losses to cattle producers in much of Africa, Central and South America, and Australia. We ini...

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

    PubMed Central

    2011-01-01

    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, Serratia marcescens, Corynebacterium glutamicum, and Finegoldia magna. One hundred twenty-one bacterial genera were detected in all the life stages and tissues sampled. The total number of genera identified by tick sample comprised: 53 in adult males, 61 in adult females, 11 in gut tissue, 7 in ovarian tissue, and 54 in the eggs. Notable genera detected in the cattle tick include Wolbachia, Coxiella, and Borrelia. The molecular approach applied in this study allowed us to assess the relative abundance of the microbiota associated with R. microplus. Conclusions This report represents the first survey of the bacteriome in the cattle tick using non-culture based molecular approaches. Comparisons of our results with previous bacterial surveys provide an indication of geographic variation in the assemblages of bacteria associated with R. microplus. Additional reports on the identification of new bacterial species maintained in nature by R. microplus that may be pathogenic to its vertebrate hosts

  7. The mitochondrial genome of a Texas outbreak strain of the cattle tick, Rhipicephalus (Boophilus) microplus, derived from whole genome sequencing Pacific Biosciences and Illumina reads

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cattle fever tick, Rhipicephalus (Boophilus) microplus is one of the most significant medical veterinary pests in the world, vectoring several serious livestock diseases negatively impacting agricultural economies of tropical and subtropical countries around the world. We assembled the complete ...

  8. Virulence of Mexican isolates of entomopathogenic fungi upon Rhipicephalus-Boophilus microplus larvae and the efficacy of conidia formulations to reduce larval tick density under field conditions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The first objective was laboratory evaluation of the virulence of 53 Mexican isolates of fungi against larvae of Rhipicephalus (Boophilus) microplus. Thirty three isolates of Metarhizium anisopliae var. anisopliae (Metschnickoff) Sorokin (Hypocreales: Clavicipitaceae) and 20 isolates of Isaria (Paec...

  9. Allopatric speciation in ticks: genetic and reproductive divergence between geographic strains of Rhipicephalus (Boophilus) microplus

    PubMed Central

    Labruna, Marcelo B; Naranjo, Victoria; Mangold, Atilio J; Thompson, Carolina; Estrada-Peña, Agustín; Guglielmone, Alberto A; Jongejan, Frans; de la Fuente, José

    2009-01-01

    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. PMID:19243585

  10. [Construction of cDNA expression library of salivary gland from Boophilus microplus].

    PubMed

    Tian, Zhan-Cheng; Liu, Guang-Yuan; Xie, Jun-Ren; Gong, Zhen-Li

    2008-10-30

    Total RNA were isolated from salivary gland dissected from partially engorged Boophilus microplus. The mRNA was purified. A library of oligo (dT)-primed cDNA with added directional EcoR I/Hind III linkers was constructed from the purified mRNA. The constructed cDNA was ligated to the EcoR I/Hind III arms of the lambda SCREEN vector. The recombinant phage DNA was packaged by phage-marker packaging extracts, resulting in a primary cDNA library with a size of 1.38x10(6) PFU. Data showed 100% of the library were recombinant and the titer of the amplified library was 2x10(9) PFU/ml. A partial cDNA encoding cytochrome oxidase C subunit II of B. microplus was screened from the expression library with rabbit serum against B. microplus salivary gland proteins. The results is suggested that the cDNA expression library has been constructed.

  11. Control of Boophilus microplus populations in grazing cattle vaccinated with a recombinant Bm86 antigen preparation.

    PubMed

    Rodríguez, M; Penichet, M L; Mouris, A E; Labarta, V; Luaces, L L; Rubiera, R; Cordovés, C; Sánchez, P A; Ramos, E; Soto, A

    1995-04-01

    Current methods for the control of cattle tick Boophilus microplus infestations are not effective and the parasite remains a serious problem for the cattle industry in tropical and subtropical areas. Recently, we developed a vaccine against B. microplus employing a recombinant Bm86 (rBm86) antigen preparation (Gavac, Heber Biotec) and it was shown to induce a protective response in vaccinated animals under controlled conditions. Here we show that, under field conditions in grazing cattle, the vaccine is able to control B. microplus populations. Two parasite-free farms were employed for the study. In the first farm, animals were vaccinated with the recombinant vaccine, while, in the second, animals received a saline injection in adjuvant. After immunization, animals were artificially infected and the infestation rate was recorded. Over the 33 weeks of the experiment, the infestation rate was lower in the vaccinated group compared with the control group. At the end of the experiment it was necessary to use chemicals in the control farm after serious losses in production and animals.

  12. Distribution Patterns of Three Sodium Channel Mutations Associated with Pyrethroid Resistance in Rhipicephalus (Boophilus) Microplus Populations from North and South America, South Africa and Australia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Resistance to synthetic pyrethroids (SP) in the cattle tick Rhipicephalus (Boophilus) microplus is widespread throughout its distribution area. Three single nucleotide substitutions identified in the Domains II and III of the sodium channel gene of R. (B.) microplus are known to be associated with t...

  13. Proteomic analysis of cattle tick Rhipicephalus (Boophilus) microplus saliva: a comparison between partially and fully engorged females.

    PubMed

    Tirloni, Lucas; Reck, José; Terra, Renata Maria Soares; Martins, João Ricardo; Mulenga, Albert; Sherman, Nicholas E; Fox, Jay W; Yates, John R; Termignoni, Carlos; Pinto, Antônio F M; Vaz, Itabajara da Silva

    2014-01-01

    The cattle tick Rhipicephalus (Boophilus) microplus is one of the most harmful parasites affecting bovines. Similarly to other hematophagous ectoparasites, R. microplus saliva contains a collection of bioactive compounds that inhibit host defenses against tick feeding activity. Thus, the study of tick salivary components offers opportunities for the development of immunological based tick control methods and medicinal applications. So far, only a few proteins have been identified in cattle tick saliva. The aim of this work was to identify proteins present in R. microplus female tick saliva at different feeding stages. Proteomic analysis of R. microplus saliva allowed identifying peptides corresponding to 187 and 68 tick and bovine proteins, respectively. Our data confirm that (i) R. microplus saliva is complex, and (ii) that there are remarkable differences in saliva composition between partially engorged and fully engorged female ticks. R. microplus saliva is rich mainly in (i) hemelipoproteins and other transporter proteins, (ii) secreted cross-tick species conserved proteins, (iii) lipocalins, (iv) peptidase inhibitors, (v) antimicrobial peptides, (vii) glycine-rich proteins, (viii) housekeeping proteins and (ix) host proteins. This investigation represents the first proteomic study about R. microplus saliva, and reports the most comprehensive Ixodidae tick saliva proteome published to date. Our results improve the understanding of tick salivary modulators of host defense to tick feeding, and provide novel information on the tick-host relationship.

  14. First report of fipronil resistance in Rhipicephalus (Boophilus) microplus of Mexico.

    PubMed

    Miller, Robert J; Almazán, Consuelo; Ortíz-Estrada, Martín; Davey, Ronald B; George, John E; De León, Adalberto Peréz

    2013-01-16

    Five strains of Rhipicephalus (Boophilus) microplus collected from Tamaulipas Mexico were tested for resistance against several classes of acaricides commonly used. All were resistant to fipronil. Four of five were co-resistant to permethrin and coumaphos in addition to being resistant to fipronil. One strain, El Zamora was found multi-resistant to permethrin, coumaphos, fipronil, and amitraz. Selection with fipronil for 3 generations produced a resistance ratio of 8.3 and 9.4 at the LC(50) and the LC(99) estimates, respectively. Permethrin resistance in El Zamora was possibly linked to elevated esterase (CZEST9) and could be a contributing factor of resistance to fipronil. The implications of resistance for the control of the southern cattle tick in the future are discussed.

  15. [Alcoholic extract of lemongrass (Cymbopogon citratus) on the control of Boophilus microplus in cattle].

    PubMed

    Heimerdinger, Arli; Olivo, Clair J; Molento, Marcelo B; Agnolin, Carlos A; Ziech, Magnos F; Scaravelli, Luciene Fernanda B; Skonieski, Fernando R; Both, José F; Charão, Pablo S

    2006-01-01

    The objective of this study was to determine the effect of lemongrass (Cymbopogon citratus) alcoholic extracts on the control of Boophilus microplus in naturally infested Holstein cows. Twelve animals were allocated in three groups of four animals. Group 1 was treated with amitraz at 0.025%, Group 2 was treated with lemongrass extracts at 1.36% and Group 3 with the same product at 2.72% of the plant. Engorged ticks were evaluated on animals with length superior to 4.0 mm, before (mean of days -3, -2, -1) and at 1, 2, 3, 4, 5, 6, 7 and 14 days after treatment. The mean efficacy of amitraz was 97.93%. Lemongrass extract at 2.72% reduced tick infestation by 40.3, 46.6 and 41.5% on day 3, 7 and 14 post-treatment, respectively.

  16. The complexity of Rhipicephalus (Boophilus) microplus genome characterised through detailed analysis of two BAC clones

    PubMed Central

    2011-01-01

    Background Rhipicephalus (Boophilus) microplus (Rmi) a major cattle ectoparasite and tick borne disease vector, impacts on animal welfare and industry productivity. In arthropod research there is an absence of a complete Chelicerate genome, which includes ticks, mites, spiders, scorpions and crustaceans. Model arthropod genomes such as Drosophila and Anopheles are too taxonomically distant for a reference in tick genomic sequence analysis. This study focuses on the de-novo assembly of two R. microplus BAC sequences from the understudied R microplus genome. Based on available R. microplus sequenced resources and comparative analysis, tick genomic structure and functional predictions identify complex gene structures and genomic targets expressed during tick-cattle interaction. Results In our BAC analyses we have assembled, using the correct positioning of BAC end sequences and transcript sequences, two challenging genomic regions. Cot DNA fractions compared to the BAC sequences confirmed a highly repetitive BAC sequence BM-012-E08 and a low repetitive BAC sequence BM-005-G14 which was gene rich and contained short interspersed elements (SINEs). Based directly on the BAC and Cot data comparisons, the genome wide frequency of the SINE Ruka element was estimated. Using a conservative approach to the assembly of the highly repetitive BM-012-E08, the sequence was de-convoluted into three repeat units, each unit containing an 18S, 5.8S and 28S ribosomal RNA (rRNA) encoding gene sequence (rDNA), related internal transcribed spacer and complex intergenic region. In the low repetitive BM-005-G14, a novel gene complex was found between to 2 genes on the same strand. Nested in the second intron of a large 9 Kb papilin gene was a helicase gene. This helicase overlapped in two exonic regions with the papilin. Both these genes were shown expressed in different tick life stage important in ectoparasite interaction with the host. Tick specific sequence differences were also

  17. Assessment of different Lippia sidoides genotypes regarding their acaricidal activity against Rhipicephalus (Boophilus) microplus.

    PubMed

    Soares, Alexandra Martins Dos Santos; Penha, Tatiane Aranha; Araújo, Sandra Alves de; Cruz, Elizangela Mércia Oliveira; Blank, Arie Fitzgerald; Costa-Junior, Livio Martins

    2016-01-01

    The aim of this study was to select different genotypes of Lippia sidoides with the highest activity against larvae and engorged females of the tick Rhipicephalus (Boophilus) microplus. The germplasms studied were LSID006, LSID102, LSID103 and LSID104. The LSID104 genotype, that presented carvacrol as a major constituent, was the germplasm with the worst larvicide effect (LC50 2.99 mg/mL). The LSID006 genotype was the least effective against engorged females (LC50 12.46 mg/mL), and it was chemically similar to the LSID102 genotype, which presented the highest acaricide activity (LC50 2.81 mg/mL). We conclude that chemical similarity of the germplasms was not correlated with acaricide activity against R. (B.) microplus. The essential oil of L. sidoides is a potent natural agent. However, the findings of this work provide a better understanding for product development based on this natural product, which must consider synergic effects or the action of minor compounds.

  18. Acaricidal activity of aqueous extract and synthesized silver nanoparticles from Manilkara zapota against Rhipicephalus (Boophilus) microplus.

    PubMed

    Rajakumar, G; Abdul Rahuman, A

    2012-08-01

    Traditional parasite control is primarily based on the use of chemical acaricides, which unfortunately have many negative side effects. The aim of the present study was to evaluate the effect of plant synthesized silver nanoparticles (AgNPs) using aqueous leaf extract of Manilkara zapota to control Rhipicephalus (Boophilus) microplus. The synthesized AgNPs were characterized by UV-vis spectrum, scanning electron microscopy (SEM), Fourier transform infrared and X-ray diffraction. The UV-vis spectrum of the aqueous medium containing silver nanostructures showed a peak at 421 nm corresponding to the surface plasmon resonance band of AgNPs. SEM supports the biosynthesis and characterization of AgNPs with spherical and oval in shape and size of 70-140 nm. Acaricidal activity of aqueous leaf extract of M. zapota and synthesized AgNPs were carried out against R. (B.) microplus and the results showed the LC(50) values of 16.72 and 3.44 mg/L; r(2)=0.856 and 0.783), respectively.

  19. Exploring the midgut proteome of partially fed female cattle tick (Rhipicephalus (Boophilus) microplus).

    PubMed

    Kongsuwan, Kritaya; Josh, Peter; Zhu, Ying; Pearson, Roger; Gough, Joanne; Colgrave, Michelle L

    2010-02-01

    The continued development of effective anti-tick vaccines remains the most promising prospect for the control of the cattle tick, Rhipicephalus (Boophilus) microplus. A vaccine based on midgut proteins could interfere with successful tick feeding and additionally interfere with midgut developmental stages of Babesia parasites, providing opportunities for the control of both the tick and the pathogens it transmits. Midgut proteins from partially fed adult female cattle ticks were analysed using a combination of 2-DE and gel-free LC-MS/MS. Analysis of the urea-soluble protein fraction resulted in the confident identification of 105 gut proteins, while the PBS-soluble fraction yielded an additional 37 R. microplus proteins. The results show an abundance of proteins involved in mitochondrial ATP synthesis, electron transport chain, protein synthesis, chaperone, antioxidant and protein folding and transport activities in midgut tissues of adult female ticks. Among the novel products identified were clathrin-adaptor protein, which is involved in the assembly of clathrin-coated vesicles, and membrane-associated trafficking proteins such as syntaxin 6 and surfeit 4. The observations allow the formulation of hypotheses regarding midgut physiology and will serve as a basis for future vaccine development and tick-host interaction research.

  20. Cattle ticks in Cameroon: is Rhipicephalus (Boophilus) microplus absent in Cameroon and the Central African region?

    PubMed

    Awa, D N; Adakal, H; Luogbou, N D D; Wachong, K H; Leinyuy, I; Achukwi, M D

    2015-03-01

    In most parts of the world, ticks are rapidly developing resistance to commonly used acaricides thus rendering control difficult. This constraint is further compounded by the introduction of new species in areas where they did not exist before. Such is the case with the introduction into and rapid spread of Rhipicephalus (Boophilus) microplus in some countries of West Africa. With the looming threat of its further spread in the region, the objective of the present study was to update knowledge on cattle ticks in Cameroon. Among 19,189 ticks collected monthly from 60 animals in 5 herds from March 2012 to February 2013, Rh. (B.) decoloratus was the most abundant species with a relative prevalence of 62.2%, followed by Amblyomma variegatum (28.4%), Rh. (B.) annulatus (0.2%), Rh. (B.) geigyi (0.03%), other Rhipicephalus spp. (8.4%) and Hyalomma spp. (0.3%). Rh. (B.) decoloratus and A. variegatum were also the most widely distributed in space. Infestation rate was generally high, with average tick count/animal of about 80 during peak periods. Tick distribution and abundance in the different sites was as varied as the underlying factors, among which the most important were management systems and climatic factors. The effects of rainfall and temperature were confounded by other factors and difficult to evaluate. However, it appears tick development depends among other factors, on a humidity threshold, above which there is not much more effect. Rh. microplus was not found during this study, but more extensive tick collections have to be done to confirm this. In conclusion, cattle tick infestation in Cameroon remains an important cause for concern. Farmers need assistance in the use and management of acaricides in order to increase their efficiency and reduce the development of resistance. Although Rh. microplus was not found, its introduction from other West African countries is imminent if adequate measures, especially in the control and limitation of animal movements

  1. In vitro activity of pineapple extracts (Ananas comosus, Bromeliaceae) on Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Domingues, Luciana Ferreira; Giglioti, Rodrigo; Feitosa, Karina Alves; Fantatto, Rafaela Regina; Rabelo, Márcio Dias; Oliveira, Márcia Cristina de Sena; Oliveira, Gilson Pereira de; Bechara, Gervasio Henrique; Chagas, Ana Carolina de Souza

    2013-07-01

    Measures to control the cattle tick, Rhipicephalus (Boophilus) microplus, based only on chemical products are becoming unsustainable, mainly because of the development of resistance. The objective of this study was to test the effect of the aqueous extract of pineapple skin (AEPS) and bromelain extracted from the stem (Sigma-Aldrich®, B4882) on engorged females and larvae of R. (B.) microplus in vitro. These substances were diluted in water and evaluated at eight concentrations. Engorged females were collected and distributed in groups of 10, with three repetitions for each treatment. After immersion in the solutions, the females were placed in an incubator for observation of survival, oviposition and larval hatching. The larval packet method was used, also with three repetitions with about 100 larvae each. The packets were incubated and the readings were performed after 24 h. The estimated reproduction and efficacy of the solutions were calculated. The LC(50) and LC(90) were estimated using the Probit procedure of the SAS program. The eight concentrations were compared within each treatment by the Tukey test. For the experiment with engorged females, the most effective concentrations were 125, 250 and 500 mg/mL: 33%, 48% and 59% for the AEPS and 27%, 51% and 55% for the bromelain. The LC(50) and LC(90) values were, respectively, 276 and 8691 mg/mL for AEPS and 373 and 5172 mg/mL for bromelain. None of the dilutions tested was effective against the larvae of R. (B.) microplus. This is the first report of the action of pineapple extracts or their constituents on cattle ticks. The results demonstrate that further studies regarding composition of tick cuticle, with evaluation of other solvents and formulations, should be conducted seeking to enhance the effect of pineapple extracts and compounds against this ectoparasite.

  2. Egg wax from the cattle tick Rhipicephalus (Boophilus) microplus inhibits Pseudomonas aeruginosa biofilm.

    PubMed

    Zimmer, Karine R; Macedo, Alexandre J; Nicastro, Gianlucca G; Baldini, Regina L; Termignoni, Carlos

    2013-09-01

    Rhipicephalus (Boophilus) microplus is constantly challenged during its life cycle by microorganisms present in their hosts or in the environment. Tick eggs may be especially vulnerable to environmental conditions because they are exposed to a rich and diverse microflora in the soil. Despite being oviposited in such hostile sites, tick eggs remain viable, suggesting that the egg surface has defense mechanisms against opportunistic and/or pathogenic organisms. R. microplus engorged females deposit a superficial wax layer onto their eggs during oviposition. This egg wax is essential for preventing desiccation as well as acting as a barrier against attack by microorganisms. In this study, we report the detection of anti-biofilm activity of R. microplus egg wax against Pseudomonas aeruginosa PA14. Genes involved in the functions of production and maintenance of the biofilm extracellular matrix, pelA and cdrA, respectively, were markedly downregulated by a tick egg-wax extract. Moreover, this extract strongly inhibited fliC gene expression. Instead of a compact extracellular matrix, P. aeruginosa PA14 treated with egg-wax extract produces a fragile one. Also, the colony morphology of cells treated with egg-wax extract appears much paler and brownish, instead of the bright purple characteristic of normal colonies. Swarming motility was also inhibited by treatment with the egg-wax extract. The inhibition of P. aeruginosa biofilm does not seem to depend on inhibition of the quorum sensing system since mRNA levels of the 3 regulators of this system were not inhibited by egg-wax extract.

  3. A comparison of three different methodologies for evaluating Rhipicephalus (Boophilus) microplus susceptibility to topical spray compounds.

    PubMed

    Corrêa, Rafael Rodrigues; Lopes, Welber Daniel Zanetti; Teixeira, Weslen Fabricio Pires; Cruz, Breno Cayeiro; Gomes, Lucas Vinicius Costa; Felippelli, Gustavo; Maciel, Willian Giquelin; Fávero, Flávia Carolina; Buzzulini, Carolina; Bichuette, Murilo Abud; Soares, Vando Edésio; da Costa, Alvimar José

    2015-01-15

    This study aimed to compare three different methodologies (Adult Immersion Tests, field trials with naturally infected animals, and a Stall Test using artificially infested cattle) to evaluate the efficacy of two topical formulations that we administered as whole body sprays (15% Cypermethrin+30% Chlorpyriphos+15% Fenthion-Colosso(®) FC 30, Ouro Fino Agronegócios; and 60% Dichlorvos+20% Chlorpyriphos-Ectofós(®), Vallée Saúde Animal Ltd.), against a susceptible strain of Rhipicephalus (Boophilus) microplus. To achieve this objective, two natural infestation trials were conducted, as well as two artificial infestation trials (Stall Tests) and two Adult Immersion Tests (AIT). The AIT results showed that both spray formulations achieved 100% efficacy against R. (B.) microplus fully engorged females. However, when observing results obtained by field trials (natural infestations) and Stall Tests, none of these topically applied compounds reached 100% efficacy or affected the reproductive capacity of the fully engorged female ticks. Additional studies must be conducted to compare these in vivo methodologies with different in vitro techniques, such as the Larval Packet Test. However, based on results obtained here, we can conclude that depending on the spray formulations used, the AIT can overestimate acaricidal efficacy and values of reproductive efficiency of such compounds against R (B.) microplus. Specifically, when dealing with spray formulations in the Stall Tests, the period of residual action can increase because these animals are sheltered from contact with environmental factors that might interfere with the efficacy of the products tested. It may be necessary to take in vivo trial results into consideration (such as field trials with naturally infested animals or Stall Tests) to standardize a specific in vitro assay, such as the Adult Immersion Test.

  4. Acaricidal activity of Ocimum basilicum and Spilanthes acmella against the ectoparasitic tick, Rhipicephalus (Boophilus) microplus (Arachinida: Ixodidae).

    PubMed

    Veeramani, V; Sakthivelkumar, S; Tamilarasan, K; Aisha, S O; Janarthanan, S

    2014-09-01

    The ectoparasitic tick, Rhipicephalus (Boophilus) microplus collected at various cattle farms in and around Chennai was subjected to treatment of different crude solvent extracts of leaves of Ocimum basilicum and Spilanthes acmella for acaricidal activity. Among various solvent extracts of leaves of O. basilicum and S. acmella used, chloroform extract of O. basilicum at concentrations between 6% and 10% exhibited 70% and 100% mortality of ticks when compared to control. The LC50 and LC90 values of the chloroform extract of leaves of O. basilicum treatment on the ticks after 24 h were observed as 5.46% and 7.69%. Quantitative and qualitative analysis of α- and β- carboxylesterase enzymes in the whole gut homogenate of cattle tick, R. microplus treated with chloroform extract of leaves of O. basilicum revealed higher level of activities for the enzymes. This indicated that there was an induced response in the tick, R. microplus against the toxic effects of the extract of O. basilicum.

  5. The mitochondrial genome of a Texas outbreak strain of the cattle tick, Rhipicephalus (Boophilus) microplus, derived from whole genome sequencing Pacific Biosciences and Illumina reads.

    PubMed

    McCooke, John K; Guerrero, Felix D; Barrero, Roberto A; Black, Michael; Hunter, Adam; Bell, Callum; Schilkey, Faye; Miller, Robert J; Bellgard, Matthew I

    2015-10-15

    The cattle fever tick, Rhipicephalus (Boophilus) microplus is one of the most significant medical veterinary pests in the world, vectoring several serious livestock diseases negatively impacting agricultural economies of tropical and subtropical countries around the world. In our study, we assembled the complete R. microplus mitochondrial genome from Illumina and Pac Bio sequencing reads obtained from the ongoing R. microplus (Deutsch strain from Texas, USA) genome sequencing project. We compared the Deutsch strain mitogenome to the mitogenome from a Brazilian R. microplus and from an Australian cattle tick that has recently been taxonomically designated as Rhipicephalus australis after previously being considered R. microplus. The sequence divergence of the Texas and Australia ticks is much higher than the divergence between the Texas and Brazil ticks. This is consistent with the idea that the Australian ticks are distinct from the R. microplus of the Americas.

  6. Bmcystatin, a cysteine proteinase inhibitor characterized from the tick Boophilus microplus

    SciTech Connect

    Lima, Cassia A.; Sasaki, Sergio D.; Tanaka, Aparecida S. . E-mail: Tanaka.bioq@epm.br

    2006-08-18

    The bovine tick Rhipicephalus (Boophilus) microplus is a blood-sucking animal, which is responsible for Babesia spp and Anaplasma marginale transmission for cattle. From a B. microplus fat body cDNA library, 465 selected clones were sequenced randomly and resulted in 60 Contigs. An open reading frame (ORF) contains 98 amino acids named Bmcystatin, due to 70% amino acid identity to a classical type 1 cystatin from Ixodes scapularis tick (GenBank Accession No. DQ066227). The Bmcystatin amino acid sequence analysis showed two cysteine residues, theoretical pI of 5.92 and M{sub r} of 11kDa. Bmcystatin gene was cloned in pET 26b vector and the protein expressed using bacteria Escherichia coli BL21 SI. Recombinant Bmcystatin (rBmcystatin) purified by affinity chromatography on Ni-NTA-agarose column and ionic exchange chromatography on HiTrap Q column presented molecular mass of 11kDa, by SDS-PAGE and the N-terminal amino acid sequenced revealed unprocessed N-terminal containing part of pelB signal sequence. Purified rBmcystatin showed to be a C1 cysteine peptidase inhibitor with K{sub i} value of 0.1 and 0.6nM for human cathepsin L and VTDCE (vitellin degrading cysteine endopeptidase), respectively. The rBmcystatin expression analyzed by semi-quantitative RT-PCR confirmed the amplification of a specific DNA sequence (294bp) in the fat body and ovary cDNA preparation. On the other hand, a protein band was detected in the fat body, ovary, and the salivary gland extracts using anti-Bmcystatin antibody by Western blot. The present results suggest a possible role of Bmcystatin in the ovary, even though the gene was cloned from the fat body, which could be another site of this protein synthesis.

  7. Kinetics of energy source utilization in Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) embryonic development.

    PubMed

    Campos, Eldo; Moraes, Jorge; Façanha, Arnoldo R; Moreira, Erica; Valle, Denise; Abreu, Leonardo; Manso, Pedro P A; Nascimento, Aline; Pelajo-Machado, Marcelo; Lenzi, Henrique; Masuda, Aoi; Vaz, Itabajara da Silva; Logullo, Carlos

    2006-06-15

    The present work evaluates the kinetics of utilization of the main potential energy sources throughout the embryonic developmental stages of Boophilus microplus. The embryonic development of this arthropod is completed in 21 days. Cellularization of the blastoderm occurs on the 6th day and is rapidly followed by germ band extension and segmentation, whose first signs are visible on the 7th day. Cellularization is typically a maternal-driven process, carried out by molecular determinants deposited in the oocyte during oogenesis. On the other hand, segmentation is of zygotic nature, being the consequence of the synthesis of various components by the growing embryo. The enhancement in total B. microplus RNA was observed after cellularization, corroborating the replacement of maternal-driven processes by embryonic zygotic expression. An abrupt increase in oxygen consumption was observed from cellularization until the 8th day of development. The reduction in dry weight at the same period and the susceptibility of oxygen consumption to KCN suggest that the respiration process is activated during early embryonic development. A marked decrease in total lipid content occurred between the 5th and 7th days of development, suggesting this is the main energy source for cellularization. A major reduction in carbohydrate content occurred later, between the 7th and 9th days, and it could be assigned to the morphological segmentation of the embryo. Although the total amount of proteins remains unchanged from oviposition to hatching, a 15% reduction in vitellin (VT) content was observed before cellularization, up to the 4th day after egglaying. This observation was correlated to the synthesis of new proteins needed to support early embryo development. Additional 20% of VT was consumed thereafter, mainly at the end of embryogenesis, and in this case VT is probably used as energy source to the older embryo. Altogether, these data indicate different energy sources for maternal and

  8. Genetics and mechanisms of permethrin resistance in the Santa Luiza strain of Boophilus microplus (Acari: Ixodidae).

    PubMed

    Li, Andrew Y; Davey, Ronald B; Miller, Robert J; Guerrero, Felix D; George, John E

    2008-05-01

    The Santa Luiza strain of the southern cattle tick, Boophilus microplus (Canestrini) (Acari: Ixodidae), is resistant to both permethrin and amitraz. A study was conducted at the USDA Cattle Fever Tick Research Laboratory in Texas to investigate the genetic basis of permethrin resistance with cross-mating experiments, and to determine the mechanisms of permethrin resistance through synergist bioassays and biochemical analysis of esterase profiles. The Muñoz strain, an acaricide-susceptible reference strain, was used as the susceptible parent and the Santa Luiza strain, originating in Brazil, was used as the resistant parent. The Food and Agriculture Organization larval packet test was used to measure the levels of susceptibility of larvae of the parental strains, F1, backcross, F2, and F3 generations to permethrin. Results of reciprocal crossing experiments suggested that permethrin resistance was inherited as an incomplete recessive trait. There was no significant maternal effect on larval progeny's susceptibility to permethrin in the F1 and subsequent generations. The values of the degree of dominance were estimated at -0.700 and -0.522 for the F1 larvae with resistant and susceptible female parents, respectively. Results of bioassays on larval progeny of the F1 backcrossed with the resistant parent strain and of the F2 generations suggested that one major gene was responsible for permethrin resistance in the Santa Luiza strain. Selection of F3 larvae with either permethrin or amitraz led to significantly increased resistance to both permethrin and amitraz, indicating a close linkage between genes responsible for permethrin and amitraz resistance. The possible involvement of metabolic enzymes in permethrin resistance in the Santa Luiza strain of B. microplus was dismissed by the lack of enhanced synergism by TPP or PBO, as observed in synergist bioassays, as well as by the lack of enhanced esterase activity in the Santa Luiza strain relative to the susceptible

  9. Acaricidal activity of five essential oils of Ocimum species on Rhipicephalus (Boophilus) microplus larvae.

    PubMed

    Hüe, T; Cauquil, L; Fokou, J B Hzounda; Dongmo, P M Jazet; Bakarnga-Via, I; Menut, C

    2015-01-01

    The aim of this study was to evaluate the acaricidal activity on the cattle tick Rhipicephalus (Boophilus) microplus of essential oils from three Ocimum species. Acaricidal activity of five essential oils extracted from Ocimum gratissimum L. (three samples), O. urticaefolium Roth, and O. canum Sims was evaluated on 14- to 21-day-old Rhipicephalus microplus tick larvae using larval packet test bioassay. These essential oils were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS) showing great variations of their chemical compositions according to the botanical species and even within the O. gratissimum species; the acaricidal activity of their main compounds was also evaluated. The essential oils of O. urticaefolium and O. gratissimum collected in Cameroon were the most efficient with respective LC50 values of 0.90 and 0.98%. The two essential oils obtained from O. gratissimum collected in New Caledonia were partially active at a dilution of 5% while the essential oil of O. canum collected in Cameroon showed no acaricidal activity. The chemical analysis shows five different profiles. Whereas the essential oils of O. urticaefolium from Cameroon and O. gratissimum from New Caledonia contain high amounts of eugenol (33.0 and 22.3-61.0%, respectively), 1,8-cineole was the main component of the oil of an O. canum sample from Cameroon (70.2%); the samples of O. gratissimum oils from New Caledonia are also characterized by their high content of (Z)-β-ocimene (17.1-49.8%) while the essential oil of O. gratissimum collected in Cameroon is mainly constituted by two p-menthane derivatives: thymol (30.5%) and γ-terpinene (33.0%). Moreover, the essential oil of O. urticaefolium showed the presence of elemicin (18.1%) as original compound. The tests achieved with the main compounds confirmed the acaricidal activity of eugenol and thymol with residual activity until 0.50 and 1%, respectively, and revealed the acaricidal property of elemicin

  10. Detection of Babesia bigemina in cattle of different genetic groups and in Rhipicephalus (Boophilus) microplus tick.

    PubMed

    Oliveira, M C S; Oliveira-Sequeira, T C G; Regitano, L C A; Alencar, M M; Néo, T A; Silva, A M; Oliveira, H N

    2008-08-17

    Babesia bigemina infections were investigated in four genetic groups of beef cattle and in Rhipicephalus (Boophilus) microplus engorged female ticks. Blood samples and engorged female ticks were collected from 15 cows and 15 calves from each of the following genetic groups: Nelore, Angus x Nelore, Canchim x Nelore, and Simmental x Nelore. Microscopic examination of blood smears and tick hemolymph revealed that merozoites of B. bigemina (6/60) as well as kinetes of Babesia spp. (9/549) were only detected in samples (blood and ticks, respectively) originated from calves. PCR-based methods using primers for specific detection of B. bigemina revealed 100% infection in both calves and cows, regardless the genetic group. Tick infection was detected by nested-PCR amplifications showing that the frequency of B. bigemina was higher (P<0.01) in female ticks collected from calves (134/549) than in those collected from cows (52/553). The frequency of B. bigemina was similar in ticks collected from animals, either cows or calves, of the four genetic groups (P>0.05).

  11. Immunogenic potential of Rhipicephalus (Boophilus) microplus aquaporin 1 against Rhipicephalus sanguineus in domestic dogs.

    PubMed

    Évora, Patricia Martinez; Sanches, Gustavo Seron; Guerrero, Felix David; León, Adalberto Pérez de; Bechara, Gervásio Henrique

    2017-02-16

    This study evaluated a recombinant aquaporin 1 protein of Rhipicephalus (Boophilus) microplus (RmAQP1) as antigen in a vaccine against R. sanguineus. Five dogs were immunized with RmAQP1 (10 µg) + adjuvant (Montanide) (G1), and five were inoculated with adjuvant only (G2), three times. Twenty-one days after the last immunization, animals of both groups were challenged with R. sanguineus larvae, nymphs and adults, and their biotic potential was compared. Blood samples were collected before each immunization and every 28 days after the last immunization for 10 weeks. Serum antibody titers (IgG) were assessed by ELISA. We observed that: engorgement period of adult females from G1 was 12% shorter than G2; larvae from G1 had 8.7% longer engorgement period than G2 and weighed 7.2% less; nymphs from G1 had 4.5% shorter engorgement period than G2 and weighed 3.6% less; although the antibody titers increased following the second immunization, they rapidly decreased after the third immunization. Results indicated low immunoprotection of RmAQP1 against adult R. sanguineus ticks, and possible efficacy on larvae and nymphs fed on immunized dogs. Further studies should be performed for a full evaluation of the immunoprotection of RmAQP1 against R. sanguineus infestations in dogs.

  12. Structural and biochemical characterization of a recombinant triosephosphate isomerase from Rhipicephalus (Boophilus) microplus.

    PubMed

    Moraes, Jorge; Arreola, Rodrigo; Cabrera, Nallely; Saramago, Luiz; Freitas, Daniela; Masuda, Aoi; da Silva Vaz, Itabajara; Tuena de Gomez-Puyou, Marietta; Perez-Montfort, Ruy; Gomez-Puyou, Armando; Logullo, Carlos

    2011-06-01

    Triosephosphate isomerase (TIM) is an enzyme with a role in glycolysis and gluconeogenesis by catalyzing the interconversion between glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. This enzyme has been used as a target in endoparasite drug development. In this work we cloned, expressed, purified and studied kinetic and structural characteristics of TIM from tick embryos, Rhipicephalus (Boophilus) microplus (BmTIM). The Km and Vmax of the recombinant BmTIM with glyceraldehyde 3-phosphate as substrate, were 0.47 mM and 6031 μmol min⁻¹ mg protein⁻¹, respectively. The resolution of the diffracted crystal was estimated to be 2.4 Å and the overall data showed that BmTIM is similar to other reported dimeric TIMs. However, we found that, in comparison to other TIMs, BmTIM has the highest content of cysteine residues (nine cysteine residues per monomer). Only two cysteines could make disulfide bonds in monomers of BmTIM. Furthermore, BmTIM was highly sensitive to the action of the thiol reagents dithionitrobenzoic acid and methyl methane thiosulfonate, suggesting that there are five cysteines exposed in each dimer and that these residues could be employed in the development of species-specific inhibitors.

  13. Isolation of protective antigens from the gut of Boophilus microplus using monoclonal antibodies.

    PubMed Central

    Lee, R P; Opdebeeck, J P

    1991-01-01

    Monoclonal antibodies (mAb) were produced against midgut membrane (GM) antigens of Boophilus microplus. The isotypes of these mAb were established and their specificity characterized using double diffusion and Western blotting. GM antigens solubilized by Triton X-100 were precipitated by mAb QU13, and the precipitate was then injected into cattle to test for the presence of protective antigens. Vaccinated cattle challenged with 10-day-old larval ticks showed evidence of protection with a 62% reduction in eggs produced by ticks from vaccinated cattle compared to tick eggs from control cattle. In a second vaccine-challenge experiment, the dose of precipitate was increased and greater than 99% protection was provided to these vaccinated cattle following challenge (calculated from tick egg weights compared to the control group). The solubilized antigen(s) precipitated by QU13 were subjected to SDS-PAGE separation and the calculated sizes of these molecules were greater than 200,000, 80,000, 74,000, 62,000 57,000 and less than 30,000 MW. Images Figure 1 Figure 2 PMID:1997395

  14. [Immunization with synthetic peptide anti-tick Rhipicephalus (Boophilus) microplus SBm7462 via mucosal routes].

    PubMed

    Carvalho, Gabriel D; Peconick, Ana P; Medeiros, Carla L; Vargas-V, Marlene I; Patarroyo, Joaquín H

    2008-09-01

    The mucosal immunization consists on antigen administration in these surfaces it is a not invasive method, inductor of local and systemic immune response. This work evaluated the immune response of the synthetic vaccine anti-tick Rhipicephalus (Boophilus) microplus SBm7462®, via oral and nasal routes. Were used 60 BALB/c mice, divided in three groups of 20 animals each (I: oral immunization; II: nasal immunization; III: animals not immunized). Nine and 15 days after the first and the second immunization was collected, separately for each group, T cells from the immunized animals, cultivating them per 10 days. After the incubation, were determined the percentage of cellular viability and the specific nature of this T cells, which had held as memory T cell. The averages of T cell SBm7462-reagents had been submitted to the analysis of variance and comparison for Tukey test, 5% of probability. Group II presented higher cellular proliferation "in vitro". For ELISA test, were positive only two animals in group I and eight in group II. The oral and nasal routs alternatively viable for immunization with the synthetic peptide SBm7462®, however require greater number of doses to induce responses with high levels of immunoglobulins.

  15. Structural and biochemical characterization of a recombinant triosephosphate isomerase from Rhipicephalus (Boophilus) microplus

    SciTech Connect

    Moraes, Jorge; Arreola, Rodrigo; Cabrera, Nallely; Saramago, Luiz; Freitas, Daniela; Masuda, Aoi; da Silva Vaz Jr., Itabajara; Tuena de Gomez-Puyou, Marietta; Perez-Montfort, Ruy; Gomez-Puyou, Armando; Logullo, Carlos

    2012-02-06

    Triosephosphate isomerase (TIM) is an enzyme with a role in glycolysis and gluconeogenesis by catalyzing the interconversion between glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. This enzyme has been used as a target in endoparasite drug development. In this work we cloned, expressed, purified and studied kinetic and structural characteristics of TIM from tick embryos, Rhipicephalus (Boophilus) microplus (BmTIM). The Km and Vmax of the recombinant BmTIM with glyceraldehyde 3-phosphate as substrate, were 0.47 mM and 6031 {micro}mol min{sup -1} mg protein{sup -1}, respectively. The resolution of the diffracted crystal was estimated to be 2.4 {angstrom} and the overall data showed that BmTIM is similar to other reported dimeric TIMs. However, we found that, in comparison to other TIMs, BmTIM has the highest content of cysteine residues (nine cysteine residues per monomer). Only two cysteines could make disulfide bonds in monomers of BmTIM. Furthermore, BmTIM was highly sensitive to the action of the thiol reagents dithionitrobenzoic acid and methyl methane thiosulfonate, suggesting that there are five cysteines exposed in each dimer and that these residues could be employed in the development of species-specific inhibitors.

  16. In vitro activities of plant extracts from the Brazilian Cerrado and Pantanal against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Barbosa, Carolina da Silva; Borges, Ligia Miranda Ferreira; Nicácio, José; Alves, Reginaldo Dias; Miguita, Carlos Henrique; Violante, Ivana Maria Póvoa; Hamerski, Lidilhone; Garcez, Walmir Silva; Garcez, Fernanda Rodrigues

    2013-07-01

    A total of 73 ethanol extracts from different anatomical parts of 44 plant species belonging to 24 families, native to the Mid-Western region of Brazil, were assessed in vitro for their effect on the reproductive cycle of engorged females of the cattle tick Rhipicephalus (Boophilus) microplus, using the adult immersion test. All extracts were evaluated at the concentration of 0.2 % and, among the extracts tested, the one obtained from the fruits of Guarea kunthiana (Meliaceae) proved to be highly efficacious, showing 99.1 % of product effectiveness. Extracts from other three species were shown to be moderately active, namely Nymphaea amazonum trunk (Nymphaeaceae) [51.7 %], Strychnos pseudoquina trunk (Loganiaceae) [48 %] [corrected] and Ocotea lancifolia leaves (Lauraceae) [34.5 %], while the remaining extracts were shown to be weakly active or inactive. This is the first report on the bioactivity of these species on egg production by engorged females of R. microplus.

  17. Acaricidal activity of Palicourea marcgravii, a species from the Amazon forest, on cattle tick Rhipicephalus (Boophilus) microplus.

    PubMed

    Silva, Wilson Castro; Martins, João Ricardo de Souza; Cesio, Maria Veronica; Azevedo, João Lúcio; Heinzen, Horacio; de Barros, Neiva Monteiro

    2011-06-30

    Leaves of Palicourea marcgravii were extracted successively with hexane, ethyl acetate and ethanol in order to evaluate their acaricidal activity on larvae and adult stages of the cattle tick Rhipicephalus (Boophilus) microplus. The ethyl acetate extract showed the highest bioactivity of the tested extracts, which contained 0.12% monofluoroacetic acid. On engorged female, the ethyl acetate extract showed a lethal concentration 50% - LC(50)=30.08 mg ml(-1), inhibitory concentration 50% - IC(50)=5.79 mg ml(-1) and lethal time 50% - LT(50)=4.72 days; 100% reproduction was controlled at concentrations of 50 mg ml(-1) and on larvae the ethyl acetate extract showed a LC(50)=2.46 mg ml(-1). No alkaloids were detected in any of the extracts. This is the first report on the acaricidal activity of P. marcgravii extracts against R. microplus as well as the acaricidal properties of a plant species containing monofluoroacetic acid.

  18. Effectiveness of Beauveria bassiana sensu lato strains for biological control against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in China.

    PubMed

    Sun, Ming; Ren, Qiaoyun; Guan, Guiquan; Li, Yufeng; Han, Xueqing; Ma, Chao; Yin, Hong; Luo, Jianxun

    2013-10-01

    Owing to the need to combat the spread of acaricide-resistant ticks, the development of long-term biological control has become a hot topic for tick control. In this study, we investigated the pathogenicity of three Beauveria bassiana isolates on the engorged female Rhipicephalus (Boophilus) microplus ticks using different conidial concentrations. The results showed that B. bassiana B.bAT17 was highly pathogenic against engorged R. (B.) microplus females, resulting in lethal time (LT50 and LT90) of 7.14 and 9.33 days at a concentration of 10(9)conidia/ml. R. (B.) microplus females treated with B. bassiana B.bAT17 significantly reduced the amount of ovipositioning; and most ticks died before they could begin to oviposit. Proteases and chitinases were analyzed in order to establish a screening method for identification of high virulent strains. This study has confirmed the significant pathogenic effect of entomopathogenic fungi against engorged R. (B.) microplus females in China, and further studies on the efficiency of the fungus against ticks in the field are required.

  19. Molecular cloning and characterization of a glycine-like receptor gene from the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Flores-Fernández, José Miguel; Gutiérrez-Ortega, Abel; Padilla-Camberos, Eduardo; Rosario-Cruz, Rodrigo; Hernández-Gutiérrez, Rodolfo; Martínez-Velázquez, Moisés

    2014-01-01

    The cattle tick Rhipicephalus (Boophilus) microplus is the most economically important ectoparasite affecting the cattle industry in tropical and subtropical areas around the world. The principal method of tick control has relied mainly on the use of chemical acaricides, including ivermectin; however, cattle tick populations resistant to ivermectin have recently been reported in Brazil, Mexico, and Uruguay. Currently, the molecular basis for ivermectin susceptibility and resistance are not well understood in R. microplus. This prompted us to search for potential molecular targets for ivermectin. Here, we report the cloning and molecular characterization of a R. microplus glycine-like receptor (RmGlyR) gene. The characterized mRNA encodes for a 464-amino acid polypeptide, which contains features common to ligand-gated ion channels, such as a large N-terminal extracellular domain, four transmembrane domains, a large intracellular loop and a short C-terminal extracellular domain. The deduced amino acid sequence showed around 30% identity to GlyRs from some invertebrate and vertebrate organisms. The polypeptide also contains the PAR motif, which is important for forming anion channels, and a conserved glycine residue at the third transmembrane domain, which is essential for high ivermectin sensitivity. PCR analyses showed that RmGlyR is expressed at egg, larval and adult developmental stages. Our findings suggest that the deduced receptor is an additional molecular target to ivermectin and it might be involved in ivermectin resistance in R. microplus.

  20. The mitochondrial genome of a Texas outbreak strain of the cattle tick, Rhipicephalus (Boophilus) microplus, derived from whole genome sequencing Pacific Biosciences and Illumina reads

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cattle fever tick, Rhipicephalus (Boophilus) microplus is one of the most significant medical veterinary pests in the world, vectoring several serious livestock diseases negatively impacting agricultural economies of tropical and subtropical countries around the world. In our study, we assembled...

  1. Efficacy of spinosad against acaricide-resistant and -susceptible Rhipicephalus (Boophilus) microplus and acaricide-susceptible Amblyomma americanum and Dermacentor variabilis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Various acaricide-resistant strains of Rhipicephalus (Boophilus) microplus, representative of the major resistance mechanisms found in Mexico and Brazil were exposed to spinosad using the FAO-Larval Packet Test and FAO-Adult Immersion Test (AIT). Larvae of all strains tested were found to be suscep...

  2. Microarray analysis of expressed genes from Rhipcephalus (Boophilus) microplus larvae, adult ovary, and adult female gut associated with Babesia bovis infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Boophilus ticks are vectors of Babesia bovis, the protozoan causative agent of cattle fever, a disease which is responsible for significant production losses to cattle producers in much of Africa, Central and South America and Australia. We utilized R. microplus microarrays, which contained over 13,...

  3. Acaricide Resistance in Rhipicephalus (Boophilus) microplus: Impact on Agro-Biosecurity and Cattle Trade between Mexico and the United States of America

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Animal health issues are important aspects of the bilateral partnership between Mexico and the United States of America (U.S.). Because the U.S. is free of the cattle fever ticks Rhipicephalus (Boophilus) microplus and R. (B.) annulatus, and bovine babesiosis, the widespread distribution of cattle f...

  4. Discovery, adaptation and transcriptional activity of two tick promoters: Construction of a dual luciferase reporter system for optimization of RNA interference in Rhipicephalus (Boophilus) microplus cell lines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Dual luciferase reporter systems are valuable tools for functional genomic studies, but have not previously been developed for use in tick cell culture. We evaluated expression of available luciferase constructs in tick cell cultures derived from Rhipicephalus (Boophilus) microplus, an important vec...

  5. Rhipicephalus (Boophilus) microplus strain Deutsch, 5 BAC clone sequencing, including two encoding Cytochrome P450s and one encoding CzEst9 carboxylesterase

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cattle tick, Rhipicephalus (Boophilus) microplus, has a genome over 2.4 times the size of the human genome, and with over 70% of repetitive DNA, this genome would prove very costly to sequence at today's prices and difficult to assemble and analyze. BAC clones give insight into the genome struct...

  6. Use of a mathematical model to study the control measures of the cattle tick Boophilus microplus population in New Caledonia.

    PubMed

    Beugnet, F; Chalvet-Monfray, K; Sabatier, P

    1998-06-30

    Boophilus microplus is a common cattle tick of great economic importance in various tropical and subtropical countries like New Caledonia. The proposed model describes the population dynamics of female Boophilus microplus in the absence of resistant ticks. It is a system of six difference equations which can be mathematically analyzed. The analysis of the system shows the great importance of the eigenvalue denoted by lambda1. The population of ticks increases if lambda1 < 1 and decreases if lambda1 > 1. The lambda1 eigenvalue depends, in particular, on the parasitic surviving rate and encounter rate between the larvae and the cows. The treatments decrease the parasitic surviving rate as the agronomic measures decrease the encounter rate. This model permits to quantify the conditions of treatments (or of the efficacy of a vaccine) and of agronomic measures by which the populations are controlled. It shows that the different treatment rhythms and the presence or not of the wild or domestic refuges plays a major role on the dynamics of tick population.

  7. Partial characterization of an atypical family I inorganic pyrophosphatase from cattle tick Rhipicephalus (Boophilus) microplus.

    PubMed

    Costa, Evenilton P; Campos, Eldo; de Andrade, Caroline P; Façanha, Arnoldo R; Saramago, Luiz; Masuda, Aoi; Vaz, Itabajara da Silva; Fernandez, Jorge H; Moraes, Jorge; Logullo, Carlos

    2012-03-23

    The present paper presents the partial characterization of a family I inorganic pyrophosphatase from the hard tick Rhipicephalus (Boophilus) microplus (BmPPase). The BmPPase gene was cloned from the tick embryo and sequenced. The deduced amino acid sequence shared high similarity with other eukaryotic PPases, on the other hand, BmPPase presented some cysteine residues non-conserved in other groups. This pyrophosphatase is inhibited by Ca(2+), and the inhibition is antagonized by Mg(2+), suggesting that the balance between free Ca(2+) and free Mg(2+) in the eggs could be involved in BmPPase activity control. We observed that the BmPPase transcripts are present in the fat body, midgut and ovary of ticks, in two developmental stages (partially and fully engorged females). However, higher transcription amounts were found in ovary from fully engorged females. BmPPase activity was considerably abolished by the thiol reagent dithionitrobenzoic acid (DTNB), suggesting that cysteine residues are exposed in its structure. Therefore, these cysteine residues play a critical role in the structural stability of BmPPase. Molecular dynamics simulation analysis indicates that BmPPase is the first Family I PPase that could promote disulfide bonds between cysteine residues 138-339 and 167-295. Finally, we believe that these cysteine residues exposed in the BmPPase structure can play an important controlling role regarding enzyme activity, which would be an interesting mechanism of redox control. The results presented here also indicate that this enzyme can be involved in embryogenesis of this arthropod, and may be useful as a target in the development of new tick control strategies.

  8. In vitro efficacy of plant extracts and synthesized substances on Rhipicephalus (Boophilus) Microplus (Acari: Ixodidae).

    PubMed

    de Souza Chagas, Ana Carolina; de Barros, Luiz Daniel; Cotinguiba, Fernando; Furlan, Maysa; Giglioti, Rodrigo; de Sena Oliveira, Márcia Cristina; Bizzo, Humberto Ribeiro

    2012-01-01

    Herbal drugs have been widely evaluated as an alternative method of parasite control, aiming to slow development of resistance and obtain low-cost biodegradable parasiticides. This study evaluated the in vitro efficacy on Rhipicephalus (Boophilus) microplus of extracts from Carapa guianensis seed oil, Cymbopogon martinii and Cymbopogon schoenanthus leaf essential oil, and Piper tuberculatum leaf crude extract and similar synthesized substances. In the immersion test, engorged females were evaluated in five dilutions ranging from 10% to 0.030625% concentration. In the larval test on impregnated filter paper, the concentration ranged from 10% to 0.02%. The treatments and controls were done in three replicates. Chemical analysis of the oils was performed by gas chromatography. The main compounds were oleic acid (46.8%) for C. guianensis and geraniol for C. martinii (81.4%), and C. schoenanthus (62.5%). The isolated and synthesized substances showed no significant effect on larvae and adult. C. martinii and P. tuberculatum showed the best efficacy on the engorged females. The LC(50) and LC(90) were 2.93% and 6.66% and 3.76% and 25.03%, respectively. In the larval test, the LC(50) and LC(90) obtained for C. martinii, P. tuberculatum, and C. schoenanthus were 0.47% and 0.63%, 0.41% and 0.79%, 0.57% and 0.96%, respectively. The fact that geraniol is present in greater quantities in C. martinii explains its higher activity in relation to C. shoenanthus. It is necessary to validate the in vivo use of safe and effective phytoparasiticidal substances. Efforts should be focused on developing formulations that enhance the efficacy in vivo and lengthen the residual period.

  9. In vitro acaricidal activity of ethanolic and aqueous floral extracts of Calendula officinalis against synthetic pyrethroid resistant Rhipicephalus (Boophilus) microplus.

    PubMed

    Godara, R; Katoch, R; Yadav, A; Ahanger, R R; Bhutyal, A D S; Verma, P K; Katoch, M; Dutta, S; Nisa, F; Singh, N K

    2015-09-01

    Detection of resistance levels against deltamethrin and cypermethrin in Rhipicephalus (Boophilus) microplus collected from Jammu (India) was carried out using larval packet test (LPT). The results showed the presence of resistance level II and I against deltamethrin and cypermethrin, respectively. Adult immersion test (AIT) and LPT were used to evaluate the in vitro efficacy of ethanolic and aqueous floral extracts of Calendula officinalis against synthetic pyrethroid resistant adults and larvae of R. (B.) microplus. Four concentrations (1.25, 2.5, 5 and 10 %) of each extract with four replications for each concentration were used in both the bioassays. A concentration dependent mortality was observed and it was more marked with ethanolic extract. In AIT, the LC50 values for ethanolic and aqueous extracts were calculated as 9.9 and 12.9 %, respectively. The egg weight of the live ticks treated with different concentrations of the ethanolic and aqueous extracts was significantly lower than that of control ticks; consequently, the reproductive index and the percent inhibition of oviposition values of the treated ticks were reduced. The complete inhibition of hatching was recorded at 10 % of ethanolic extract. The 10 % extracts caused 100 % mortality of larvae after 24 h. In LPT, the LC50 values for ethanolic and aqueous extracts were determined to be 2.6 and 3.2 %, respectively. It can be concluded that the ethanolic extract of C. officinalis had better acaricidal properties against adults and larvae of R. (B.) microplus than the aqueous extract.

  10. Acaricidal activity of extracts from Petiveria alliacea (Phytolaccaceae) against the cattle tick, Rhipicephalus (Boophilus) microplus (Acari: ixodidae).

    PubMed

    Rosado-Aguilar, J A; Aguilar-Caballero, A; Rodriguez-Vivas, R I; Borges-Argaez, R; Garcia-Vazquez, Z; Mendez-Gonzalez, M

    2010-03-25

    The acaricidal activity of crude extracts and fractions from stems and leaves of Petiveria alliacea (Phytolaccaceae) was carried out on larvae and adults of the cattle tick Rhipicephalus (Boophilus) microplus using the larval immersion test (LIT) and adult immersion test (AIT), respectively. Methanolic extracts of stems and leaves of P. alliacea showed 100% mortality on the LIT bioassay. On the other hand, methanolic extracts of leaves and stem on the AIT test showed 26% and 86% of mortality, respectively, egg laying inhibition of 40% and 91%, respectively and hatchability inhibition of 26% and 17%, respectively. Purification of the active stem methanolic extract showed that the activity was present in the n-hexane non-polar fraction. Bioassay-guided purification of the n-hexane fraction produced 10 semi-purified fractions; fraction B had the highest activity against tick larvae (100% mortality). Gas chromatography-mass spectrometry demonstrated that the chemical composition of the active fraction B samples were mainly composed of benzyltrisulfide (BTS) and benzyldisulfide (BDS). These metabolites might be responsible for the acaricidal activity of stem extract of P. alliacea. However, further experiments to evaluate the acaricidal activity of BTS and BDS on larvae and adults of R. (B.) microplus are needed. Our results showed that P. alliacea is a promising biocontrol candidate as acaricide against R. (B.) microplus resistant strains.

  11. Integrated control of Boophilus microplus ticks in Cuba based on vaccination with the anti-tick vaccine Gavac.

    PubMed

    Valle, Manuel Rodriguez; Mèndez, Luis; Valdez, Mario; Redondo, Miguel; Espinosa, Carlos Montero; Vargas, Milagro; Cruz, Ricardo Lleonart; Barrios, Humberto Perez; Seoane, Guillermo; Ramirez, Emerio Serrano; Boue, Oscar; Vigil, Jorge Lodos; Machado, Héctor; Nordelo, Carlos Borroto; Piñeiro, Marisdania Joglar

    2004-01-01

    Boophilus microplus has developed resistance against a range of chemical acaricides which has stimulated the development of alternative methods such as vaccination against ticks. In Cuba, the Bm86-based recombinant vaccine Gavac has been successfully used in a number of controlled laboratory and field trials in cattle against B. microplus. In this paper, we have evaluated Gavac in a large scale field trial wherein 588,573 dairy cattle were vaccinated with the aim to reduce the number of acaricidal treatments. It was found that the number of acaricidal treatments could be reduced by 87% over a period of 8 years (1995--2003). Prior to the introduction of the vaccine, 54 clinical cases of babesiosis and six fatal cases were reported per 1000 animals. Six years later, the incidence of babesiosis was reduced to 1.9 cases per 1000 cattle and mortality reduced to 0.18 per 1000. The national consumption of acaricides in Cuba could be reduced by 82% after the implementation of the integrated anti-B. microplus control program.

  12. Efficacy of Rhipicephalus (Boophilus) microplus Bm86 against Hyalomma dromedarii and Amblyomma cajennense tick infestations in camels and cattle.

    PubMed

    Rodríguez-Valle, Manuel; Taoufik, Amar; Valdés, Mario; Montero, Carlos; Ibrahin, Hassan; Hassan, Shawgi Mohammed; Jongejan, Frans; de la Fuente, Jose

    2012-05-14

    The recombinant Bm86-based tick vaccines have shown their efficacy for the control of cattle ticks, Rhipicephalus (Boophilus) microplus and R. annulatus infestations. However, cattle ticks often co-exist with multi-host ticks such as Hyalomma and Amblyomma species, thus requiring the control of multiple tick infestations for cattle and other hosts. Vaccination trials using a R. microplus recombinant Bm86-based vaccine were conducted in cattle and camels against Hyalomma dromedarii and in cattle against Amblyomma cajennense immature and adult ticks. The results showed an 89% reduction in the number of H. dromedarii nymphs engorging on vaccinated cattle, and a further 32% reduction in the weight of the surviving adult ticks. In vaccinated camels, a reduction of 27% and 31% of tick engorgement and egg mass weight, respectively was shown, while egg hatching was reduced by 39%. However, cattle vaccination with Bm86 did not have an effect on A. cajennense tick infestations. These results showed that Bm86 vaccines are effective against R. microplus and other tick species but improved vaccines containing new antigens are required to control multiple tick infestations.

  13. Analysis of Babesia bovis infection-induced gene expression changes in larvae from the cattle tick, Rhipicephalus (Boophilus) microplus

    PubMed Central

    2012-01-01

    Background Cattle babesiosis is a tick-borne disease of cattle that has severe economic impact on cattle producers throughout the world’s tropical and subtropical countries. The most severe form of the disease is caused by the apicomplexan, Babesia bovis, and transmitted to cattle through the bite of infected cattle ticks of the genus Rhipicephalus, with the most prevalent species being Rhipicephalus (Boophilus) microplus. We studied the reaction of the R. microplus larval transcriptome in response to infection by B. bovis. Methods Total RNA was isolated for both uninfected and Babesia bovis-infected larval samples. Subtracted libraries were prepared by subtracting the B. bovis-infected material with the uninfected material, thus enriching for expressed genes in the B. bovis-infected sample. Expressed sequence tags from the subtracted library were generated, assembled, and sequenced. To complement the subtracted library method, differential transcript expression between samples was also measured using custom high-density microarrays. The microarray probes were fabricated using oligonucleotides derived from the Bmi Gene Index database (Version 2). Array results were verified for three target genes by real-time PCR. Results Ticks were allowed to feed on a B. bovis-infected splenectomized calf and on an uninfected control calf. RNA was purified in duplicate from whole larvae and subtracted cDNA libraries were synthesized from Babesia-infected larval RNA, subtracting with the corresponding uninfected larval RNA. One thousand ESTs were sequenced from the larval library and the transcripts were annotated. We used a R. microplus microarray designed from a R. microplus gene index, BmiGI Version 2, to look for changes in gene expression that were associated with infection of R. microplus larvae. We found 24 transcripts were expressed at a statistically significant higher level in ticks feeding upon a B. bovis-infected calf contrasted to ticks feeding on an uninfected calf

  14. In vitro acaricidal effect of tannin-rich plants against the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Fernández-Salas, A; Alonso-Díaz, M A; Acosta-Rodríguez, R; Torres-Acosta, J F J; Sandoval-Castro, C A; Rodríguez-Vivas, R I

    2011-01-10

    The objectives of this study were to evaluate the in vitro acaricidal effects of lyophilized extracts of four tannin rich plants (Acacia pennatula, Piscidia piscipula, Leucaena leucocephala and Lysiloma latisiliquum) against diverse stages of Rhipicephalus (Boophilus) microplus, and to asses whether tannins were involved in the acaricidal effect using polyethylene glycol (PEG) to block tannins. Larval immersion (LIT) and adult immersion (AIT) tests were used to evaluate the acaricidal effect of each of the lyophilized extracts against larval and adult stages of R. microplus respectively. Larvae and adult ticks were exposed to increasing concentrations of each plant extract (0, 1200, 2400, 4800, 9600 and 19,200 μg ml(-1)) for 10 min. Larval mortality was recorded at 48 h post-incubation. Adult mortality was recorded daily over 14 days, at which point their reproductive efficiency was evaluated. PEG was added to the extracts to verify whether tannins were involved in the acaricidal effect. The effect on egg laying inhibition and larval mortality was analyzed using the GLM procedure in SAS. A Kruskal-Wallis test was used to assess the effect of PEG on LIT results. Calculation of the lethal concentration 50 (LC50) was performed using a probit analysis. All extracts reduced the viability of R. microplus larval stages (P<0.001), and viability was restored with the addition of PEG suggesting an important role of tannins in the acaricidal effect (P<0.001). The LC50 values of L. latisiliquum and P. piscipula plant extracts were 6.402 and 2.466 μg ml(-1). None of the tannin-rich plant extracts affected adult mortality (P>0.05). Lysiloma latisiliquum extract inhibited egg hatching of R. microplus (P<0.01). Tannin-rich plant extracts from A. pennatula, P. piscipula, L. leucocephala and L. latisiliquum showed potential acaricidal activity. Further in vivo studies are needed to confirm this finding.

  15. Efficacy of 11 Brazilian essential oils on lethality of the cattle tick Rhipicephalus (Boophilus) microplus.

    PubMed

    Chagas, Ana Carolina de Souza; Oliveira, Márcia Cristina de Sena; Giglioti, Rodrigo; Santana, Raul Costa Mascarenhas; Bizzo, Humberto Ribeiro; Gama, Paola Ervatti; Chaves, Francisco Celio Maia

    2016-04-01

    Herbal extracts have been investigated as an alternative for parasite control, aiming to slow the development of resistance and to obtain low-cost biodegradable parasiticides. The goal of this study was to evaluate the efficacy, in vitro, of 11 essential oils from Brazil on reproductive efficiency and lethality of the cattle tick Rhipicephalus (Boophilus) microplus. The effects of oils extracted from Curcuma longa, Zingiber officinale, Lippia alba, Lippia gracilis, Lippia origanoides, Lippia sidoides, Mentha arvensis, Mentha piperita, Croton cajucara (white and red), and Croton sacaquinha on ticks were investigated by the Immersion Test with Engorged Females (ITEF) and the modified Larval Packet Test (LPT). Distilled water and 2% Tween 80 were used as control treatments. Chemical analysis of the oils was done with gas chromatography coupled with mass spectrometry. Analysis of the in vitro tests using Probit (SAS program) allowed the calculation of lethal concentrations (LCs). Lower reproductive efficiency indexes and higher efficacy percentages in the ITEF were obtained with the oils extracted from C. longa (24 and 71%, respectively) and M. arvensis oils (27 and 73%, respectively). Lower LC50 was reached with C. longa (10.24 mg/mL), L. alba (10.78 mg/mL), M. arvensis (22.31 mg/mL), L. sidoides (27.67 mg/mL), and C. sacaquinha (29.88 mg/mL) oils. In the LPT, species from Zingiberaceae and Verbenaceae families caused 100% lethality at 25 mg/mL, except for L. sidoides. The most effective oils were from C. longa, L. gracilis, L. origanoides, L. alba, and Z. officinale. The LC50 and LC90 were, respectively: 0.54 and 1.80 mg/mL, 3.21 and 7.03 mg/mL, 3.10 and 8.44 mg/mL, 5.85 and 11.14 mg/mL, and 7.75 and 13.62 mg/mL. The efficacy was directly related to the major components in each essential oil, and the oils derived from Croton genus presented the worst performance, suggesting the absence of synergistic effect among its compounds. Since C. longa, containing 62

  16. Inhibition of enzyme activity of Rhipicephalus (Boophilus) microplus triosephosphate isomerase and BME26 cell growth by monoclonal antibodies.

    PubMed

    Saramago, Luiz; Franceschi, Mariana; Logullo, Carlos; Masuda, Aoi; Vaz, Itabajara da Silva; Farias, Sandra Estrazulas; Moraes, Jorge

    2012-10-12

    In the present work, we produced two monoclonal antibodies (BrBm37 and BrBm38) and tested their action against the triosephosphate isomerase of Rhipicephalus (Boophilus) microplus (RmTIM). These antibodies recognize epitopes on both the native and recombinant forms of the protein. rRmTIM inhibition  by BrBm37 was up to 85% whereas that of BrBrm38 was 98%, depending on the antibody-enzyme ratio. RmTIM activity was lower in ovarian, gut, and fat body tissue extracts treated with BrBm37 or BrBm38 mAbs. The proliferation of the embryonic tick cell line (BME26) was inhibited by BrBm37 and BrBm38 mAbs. In summary, the results reveal that it is possible to interfere with the RmTIM function using antibodies, even in intact cells.

  17. Inhibition of Enzyme Activity of Rhipicephalus (Boophilus) microplus Triosephosphate Isomerase and BME26 Cell Growth by Monoclonal Antibodies

    PubMed Central

    Saramago, Luiz; Franceschi, Mariana; Logullo, Carlos; Masuda, Aoi; Vaz, Itabajara da Silva; Farias, Sandra Estrazulas; Moraes, Jorge

    2012-01-01

    In the present work, we produced two monoclonal antibodies (BrBm37 and BrBm38) and tested their action against the triosephosphate isomerase of Rhipicephalus (Boophilus) microplus (RmTIM). These antibodies recognize epitopes on both the native and recombinant forms of the protein. rRmTIM inhibition by BrBm37 was up to 85% whereas that of BrBrm38 was 98%, depending on the antibody-enzyme ratio. RmTIM activity was lower in ovarian, gut, and fat body tissue extracts treated with BrBm37 or BrBm38 mAbs. The proliferation of the embryonic tick cell line (BME26) was inhibited by BrBm37 and BrBm38 mAbs. In summary, the results reveal that it is possible to interfere with the RmTIM function using antibodies, even in intact cells. PMID:23202941

  18. Differential expression of genes in salivary glands of male Rhipicephalus (Boophilus)microplus in response to infection with Anaplasma marginale

    PubMed Central

    2010-01-01

    Background Bovine anaplasmosis, caused by the rickettsial tick-borne pathogen Anaplasma marginale (Rickettsiales: Anaplasmataceae), is vectored by Rhipicephalus (Boophilus)microplus in many tropical and subtropical regions of the world. A. marginale undergoes a complex developmental cycle in ticks which results in infection of salivary glands from where the pathogen is transmitted to cattle. In previous studies, we reported modification of gene expression in Dermacentor variabilis and cultured Ixodes scapularis tick cells in response to infection with A. marginale. In these studies, we extended these findings by use of a functional genomics approach to identify genes differentially expressed in R. microplus male salivary glands in response to A. marginale infection. Additionally, a R. microplus-derived cell line, BME26, was used for the first time to also study tick cell gene expression in response to A. marginale infection. Results Suppression subtractive hybridization libraries were constructed from infected and uninfected ticks and used to identify genes differentially expressed in male R. microplus salivary glands infected with A. marginale. A total of 279 ESTs were identified as candidate differentially expressed genes. Of these, five genes encoding for putative histamine-binding protein (22Hbp), von Willebrand factor (94Will), flagelliform silk protein (100Silk), Kunitz-like protease inhibitor precursor (108Kunz) and proline-rich protein BstNI subfamily 3 precursor (7BstNI3) were confirmed by real-time RT-PCR to be down-regulated in tick salivary glands infected with A. marginale. The impact of selected tick genes on A. marginale infections in tick salivary glands and BME26 cells was characterized by RNA interference. Silencing of the gene encoding for putative flagelliform silk protein (100Silk) resulted in reduced A. marginale infection in both tick salivary glands and cultured BME26 cells, while silencing of the gene encoding for subolesin (4D8

  19. The ovarian transcriptome of the cattle tick, Rhipicephalus (Boophilus) microplus, feeding upon a bovine host infected with Babesia bovis

    PubMed Central

    2013-01-01

    Background Cattle babesiosis is a tick-borne disease of cattle with the most severe form of the disease caused by the apicomplexan, Babesia bovis. Babesiosis is transmitted to cattle through the bite of infected cattle ticks of the genus Rhipicephalus. The most prevalent species is Rhipicephalus (Boophilus) microplus, which is distributed throughout the tropical and subtropical countries of the world. The transmission of B. bovis is transovarian and a previous study of the R. microplus ovarian proteome identified several R. microplus proteins that were differentially expressed in response to infection. Through various approaches, we studied the reaction of the R. microplus ovarian transcriptome in response to infection by B. bovis. Methods A group of ticks were allowed to feed on a B. bovis-infected splenectomized calf while a second group fed on an uninfected splenectomized control calf. RNA was purified from dissected adult female ovaries of both infected and uninfected ticks and a subtracted B. bovis-infected cDNA library was synthesized, subtracting with the uninfected ovarian RNA. Four thousand ESTs were sequenced from the ovary subtracted library and annotated. Results The subtracted library dataset assembled into 727 unique contigs and 2,161 singletons for a total of 2,888 unigenes, Microarray experiments designed to detect B. bovis-induced gene expression changes indicated at least 15 transcripts were expressed at a higher level in ovaries from ticks feeding upon the B. bovis-infected calf as compared with ovaries from ticks feeding on an uninfected calf. We did not detect any transcripts from these microarray experiments that were expressed at a lower level in the infected ovaries compared with the uninfected ovaries. Using the technique called serial analysis of gene expression, 41 ovarian transcripts from infected ticks were differentially expressed when compared with transcripts of controls. Conclusion Collectively, our experimental approaches provide

  20. Further studies on South African plants: Acaricidal activity of organic plant extracts against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Wellington, Kevin W; Leboho, Tlabo; Sakong, Bellonah M; Adenubi, Olubukola T; Eloff, Jacobus N; Fouche, Gerda

    2017-01-30

    The goal of our research is to develop a lower cost eco-friendly tick control method because acaricides that are commonly used to control ticks are often toxic, harmful to the environment or too expensive for resource-limited farmers. Acetone and ethanol extracts were prepared and their acaricidal activities determined against the southern cattle tick, Rhipicephalus (Boophilus) microplus. A 1% solution of each of the plant extracts was prepared for efficacy testing using the adapted Shaw Larval Immersion Test (SLIT). The acetone stem extract from Cissus quadrangularis (Vitaceae) and the ethanol leaf and flower extract from Calpurnia aurea (Fabaceae) had potent activity like that of the commercial acaricide, chlorfenvinphos [corrected mortality (CM)=100.0%]. The ethanol extracts of the stem of C. quadrangularis (CM=98.9%) and that of the roots, leaves and fruit of Senna italica subsp arachoides (CM=96.7%) also had good acaricidal activity. There is potential for the development of botanicals as natural acaricides against R. (B.) microplus that can be used commercially to protect animals against tick infestation. Further studies to isolate the acaricidal active compounds and to determine the environmental fate, species toxicity and skin toxicity of these plants species are, however, required before they can be considered as a treatment against ticks.

  1. [Selection of isolates of entomopathogenic fungus Beauveria bassiana (Ascomycetes: Clavicipitaceae) for control of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)].

    PubMed

    Barci, Leila A G; de Almeida, José Eduardo M; de Campos Nogueira, Adriana H; Zappelini, Luciano O; do Prado, Angelo P

    2009-12-01

    This study was carried out to select isolates of the entomopathogenic fungus Beauveria bassiana with pathogenic potential to control the Rhipicephalus (Boophilus) microplus tick. The effectiveness of thirty isolates was first tested at a concentration of 5 x 108 conidia.mL(-1). Of these, eight were evaluated (IBCB01, IBCB02, IBCB07, IBCB17, IBCB21, IBCB74, IBCB149, IBCB165) and showed an effectiveness between 90 and 99%; thirteen (IBCB03, IBCB14, IBCB16, IBCB24, IBCB95, IBCB97, IBCB102, IBCB141, IBCB146, IBCB147, IBCB150, IBCB154, IBCB157) between 80 and 89,5%; six (IBCB47, IBCB75, IBCB84, IBCB145, IBCB161, IBCB164) between 70 and 79%, and only two (IBCB13 and IBCB143) had lower pathogenicity (70% or below). In the second step of the study, the five more effective strains in the first phase of the experiment (IBCB01, IBCB07, IBCB21, IBCB66, IBCB165) were analyzed comparatively. Based on in vitro results, it can be concluded that IBCB66 and IBCB21 are the isolates with higher potential for field control of R. (B.) microplus. IBCB01, IBCB07, IBCB21, IBCB66 e IBCB165 isolates were submitted to a conidial production test using a rice-based substrate. The best mass production of the entomopathogenic fungus was obtained with the IBCB66 strain.

  2. Immune response in mice and cattle after immunization with a Boophilus microplus DNA vaccine containing bm86 gene.

    PubMed

    Ruiz, Lina María; Orduz, Sergio; López, Elkin D; Guzmán, Fanny; Patarroyo, Manuel E; Armengol, Gemma

    2007-03-15

    Plasmid pBMC2 encoding antigen Bm86 from a Colombian strain of cattle tick Boophilus microplus, was used for DNA-mediated immunization of BALB/c mice, employing doses of 10 and 50microg, delivered by intradermic and intramuscular routes. Anti-Bm86 antibody levels were significantly higher compared to control mice treated with PBS. In the evaluation of immunoglobulin isotypes, significant levels of IgG2a and IgG2b were observed in mice immunized with 50microg of pBMC2. Measurement of interleukine (IL) levels (IL-4, IL-5, IL-12(p40)) and interferon-gamma (IFN-gamma) in the sera of mice immunized with pBMC2 indicated high levels of IL-4 and IL-5, although there were also significant levels of IFN-gamma. Mice immunized with pBMC2 showed antigen-specific stimulation of splenocytes according to the incorporation of bromodeoxyuridine and IFN-gamma secretion. In all trials, mice injected intramuscularly with 50microg of pBMC2 presented the highest immune response. Moreover, cattle immunized with this DNA vaccine showed antibody production significantly different to the negative control. In conclusion, these results suggest the potential of DNA immunization with pBMC2 to induce humoral and cellular immune responses against B. microplus.

  3. Anaplasma marginale infection in a Japanese Black cow 13 years after eradication of Rhipicephalus (Boophilus) microplus in Okinawa, Japan.

    PubMed

    Ooshiro, Mamoru; Zakimi, Satoshi; Matsukawa, Yoshimasa; Yafuso, Makoto; Katagiri, Yoshito; Inokuma, Hisashi

    2009-03-23

    In October 2007, a 15-year-old Japanese Black cow on Ishigaki Island, Okinawa, Japan, was diagnosed with Anaplasma marginale infection based on clinical symptoms, blood examination, smear observation, 16S rRNA and groEL gene sequence analysis, and the result of a CF test. The cow was introduced into the farm from mainland Japan as a calf in 1993, one year before the eradication of Rhipicephalus (Boophilus) microplus, the main vector of A. marginale in Okinawa Prefecture. It is possible that the cow was first infected with A. marginale as a calf in Ishigaki Island and had been persistently infected since then. This is the first reported clinical case of A. marginale infection of cattle since the eradication of R. microplus in Okinawa Prefecture. Additional analysis of major surface protein 1alpha amino acid sequences revealed that the A. marginale Okinawa strain presented four new repeat forms which were not seen in other strains. This indicates that the Okinawa strain may be a unique geographical variant of A. marginale.

  4. First report of ivermectin resistance in field populations of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in Punjab districts of India.

    PubMed

    Singh, Nirbhay Kumar; Singh, Harkirat; Jyoti; Prerna, Mranalini; Rath, Shitanshu S

    2015-11-30

    The larval immersion test (LIT) was used on the progenies of Rhipicephalus (Boophilus) microplus collected from four districts of the Punjab state, India to test the resistance to ivermectin. The regression graphs of probit mortality of larval ticks plotted against log values of increasing concentrations of ivermectin were utilized for the determination of slope of mortality, lethal concentration for 50%(LC50), 95%(LC95), resistance ratios (RR50, RR95) and the resistance levels (RL). Values of the coefficient of determination (R(2)) for LIT ranged from 0.900 to 0.978, and the RR50 and RR95 values against ivermectin ranged from 2.97 to 8.85 and 2.42 to 8.47, respectively, indicating resistance status in all field isolates. On the basis of RR values, three field isolates (BAT, GUR, HOS) showed level II, whereas PTK isolate showed presence of level I resistance status against ivermectin. This appears to be the first report of ivermectin resistance in R. (B.) microplus from Punjab, India.

  5. Identification of potential plant extracts for anti-tick activity against acaricide resistant cattle ticks, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Ghosh, Srikanta; Tiwari, Shashi Shankar; Kumar, Bhanu; Srivastava, Sharad; Sharma, Anil Kumar; Kumar, Sachin; Bandyopadhyay, A; Julliet, Sanis; Kumar, Rajesh; Rawat, A K S

    2015-05-01

    To develop an eco-friendly tick control method, seven plant extracts were prepared using 50 and 95% ethanol and evaluated for acaricidal activity against cattle tick, Rhipicephalus (Boophilus) microplus. The adult immersion test was adopted for testing different extracts. Based on 72 h screening criterion, 95% ethanolic extracts of Datura metel fruits and Argemone mexicana whole plant were found effective showing more than 50% mortality of treated ticks. The 95% ethanolic extracts of D. metel fruits and A. mexicana whole plant exhibited acaricidal and reproductive inhibitory effects on treated ticks. The LC90 values of D. metel and A. mexicana extracts were determined as 7.13 and 11.3%, respectively. However, although both the extracts were found efficacious against deltamethrin-resistant IVRI-4 and multi-acaricide resistant IVRI-5 lines of R. (B.) microplus, they caused less mortality than treated ticks of the reference IVRI-I line. Phytochemical studies indicated the presence of alkaloids and glucosides in D. metel fruits and alkaloids, terpenoids, flavonoids and phenolics in A. mexicana whole plant extracts. The results indicated that these botanicals may play an important role in reducing the use of chemicals for tick control and possibly to manage resistant tick population in environment friendly manner.

  6. Acaricidal activity of essential oils from five endemic conifers of New Caledonia on the cattle tick Rhipicephalus (Boophilus) microplus.

    PubMed

    Lebouvier, Nicolas; Hue, Thomas; Hnawia, Edouard; Lesaffre, Leïla; Menut, Chantal; Nour, Mohammed

    2013-04-01

    The aim of the present study was to demonstrate acaricidal activity on the cattle tick Rhipicephalus (Boophilus) microplus of essential oils from endemic conifers of New Caledonia in the context of the development of natural alternatives. Acaricidal activity of essential oils extracted from resin and heartwood of five endemic conifers of New Caledonia (Araucaria columnaris, Agathis moorei, Agathis ovata, Callitris sulcata, and Neocallitropsis pancheri) was evaluated on 14- to 21-day-old Rhipicephalus microplus tick larvae using the Larval Packal Test bioassay. A first screening with 5% dilute solution was carried out and the oils with 100% of mortality at this rate were diluted until no activity was shown. The heartwood oils of the two Cupressaceae were the most active with LC50 value of 0.65% for C. sulcata and 0.55% for N. pancheri while resin oil of A. columnaris (LC50=1.62%) was the most active of the Araucariaceae family. Negative control (ethanol) was not toxic to the larvae. The chemical composition of essential oil from resin of A. columnaris was analyzed by gas chromatography-mass spectrometry. The essential oil was characterized by high level of sesquiterpene hydrocarbons and oxygenated sesquiterpenes and was composed mainly of aromadendrene (23.1%) and bicyclogermacrene (16.0%). In order to compare different plant resources in a sustainable program of natural acaricide, an "essential oil efficiency EOE" can be measured as the ratio between the yield of extraction and LC50 value. This study shows that A. columnaris (EOE=2.36) and N. pancheri (EOE=3.51) could provide valuable and effective natural acaricides for control of the cattle tick R. microplus.

  7. Identification of immunogenic proteins from ovarian tissue and recognized in larval extracts of Rhipicephalus (Boophilus) microplus, through an immunoproteomic approach.

    PubMed

    Ramírez Rodríguez, Patricia Berenice; Rosario Cruz, Rodrigo; Domínguez García, Delia Inés; Hernández Gutiérrez, Rodolfo; Lagunes Quintanilla, Rodolfo Esteban; Ortuño Sahagún, Daniel; González Castillo, Celia; Gutiérrez Ortega, Abel; Herrera Rodríguez, Sara Elisa; Vallejo Cardona, Adriana; Martínez Velázquez, Moisés

    2016-11-01

    Rhipicephalus (Boophilus) microplus ticks are obligatory hematophagous ectoparasites of cattle and act as vectors for disease-causing microorganisms. Conventional tick control is based on the use of chemical acaricides; however, their uncontrolled use has increased tSresistant tick populations, as well as food and environmental contamination. Alternative immunological tick control has shown to be partially effective. The only anti-tick vaccine commercially available at present in the world is based on intestinal Bm86 protein, and shows a variable effectiveness depending on tick strains or geographic isolates. Therefore, there is a need to characterize new antigens in order to improve immunological protection. The aim of this work was to identify immunogenic proteins from ovarian tissue extracts of R. microplus, after cattle immunization. Results showed that ovarian proteins complexed with the adjuvant Montanide ISA 50 V generated a strong humoral response on vaccinated cattle. IgG levels peaked at fourth post-immunization week and remained high until the end of the experiment. 1D and 2D SDS-PAGE-Western blot assays with sera from immunized cattle recognized several ovarian proteins. Reactive bands were cut and analyzed by LC-MS/MS. They were identified as Vitellogenin, Vitellogenin-2 precursor and Yolk Cathepsin. Our findings along with bioinformatic analysis indicate that R. microplus has several Vitellogenin members, which are proteolytically processed to generate multiple polypeptide fragments. This apparent complexity of vitellogenic tick molecular targets gives the opportunity to explore their potential usefulness as vaccine candidates but, at the same time, imposes a challenge on the selection of the appropriate set of antigens.

  8. Resistance to coumaphos and diazinon in Boophilus microplus (Acari: Ixodidae) and evidence for the involvement of an oxidative detoxification mechanism.

    PubMed

    Li, Andrew Y; Davey, Ronald B; Miller, Robert J; George, John E

    2003-07-01

    The levels of resistance to two organophosphate acaricides, coumaphos and diazinon, in several Mexican strains of Boophilus microplus (Canestrini) were evaluated using the FAO larval packet test. Regression analysis of LC50 data revealed a significant cross-resistance pattern between those two acaricides. Metabolic mechanisms of resistance were investigated with synergist bioassays. Piperonyl butoxide (PBO) reduced coumaphos toxicity in susceptible strains, but synergized coumaphos toxicity in resistant strains. There was a significant correlation between PBO synergism ratios and the coumaphos resistance ratios. The results suggest that an enhanced cytochrome P450 monooxygenase (cytP450)-mediated detoxification mechanism may exist in the resistant strains, in addition to the cytP450-mediated metabolic pathway that activates coumaphos. PBO failed to synergize diazinon toxicity in resistant strains, suggesting the cytP450 involved in detoxification were specific. Triphenylphosphate (TPP) synergized toxicity of both acaricides in both susceptible and resistant strains, and there was no correlation between TPP synergism ratios and the LC50 estimates for either acaricide. Esterases may not play a major role in resistance to coumaphos and diazinon in those strains. Bioassays with diethyl maleate (DEM) revealed a significant correlation between DEM synergism ratios and LC50 estimates for diazinon, suggesting a possible role for glutathione S-transferases in diazinon detoxification. Resistance to coumaphos in the Mexican strains of B. microplus was likely to be conferred by both a cytP450-mediated detoxification mechanism described here and the mechanism of insensitive acetylcholinesterases reported elsewhere. The results of this study also underscore the potential risk of coumaphos resistance in B. microplus from Mexico to the U.S. cattle fever tick eradication program.

  9. Integrated control of acaricide-resistant Boophilus microplus populations on grazing cattle in Mexico using vaccination with Gavac and amidine treatments.

    PubMed

    Redondo, M; Fragoso, H; Ortíz, M; Montero, C; Lona, J; Medellín, J A; Fría, R; Hernández, V; Franco, R; Machado, H; Rodríguez, M; de la Fuente, J

    1999-10-01

    Throughout most of the twentieth century, tick infestations on cattle have been controlled with chemical acaricides, typically administered by dipping or spraying. This approach can cause environmental and residue problems and has created a high incidence of acaricide resistance within tick populations in the field. Recently we developed a vaccine against Boophilus microplus employing a recombinant Bm86 antigen preparation (Gavac), (Heber Biotec S.A., Havana, Cuba) which has been shown to induce a protective response in vaccinated animals. Here we show for the first time under field conditions a near 100% control of B. microplus populations resistant to pyrethroids and organophosphates, by an integrated system employing vaccination with Gavac and amidine treatments. This method effectively controls tick infestations while reducing the number of chemical acaricide treatments and consequently the rise of B. microplus populations resistant to chemical acaricides.

  10. In vitro evaluation of acaricidal activity of novel green silver nanoparticles against deltamethrin resistance Rhipicephalus (Boophilus) microplus.

    PubMed

    Avinash, B; Venu, R; Alpha Raj, M; Srinivasa Rao, K; Srilatha, Ch; Prasad, T N V K V

    2017-04-15

    An investigation was undertaken to study, for the first time, in vitro acaricidal activity of green silver nanoparticles on deltamethrin resistance Rhipicephalus (Boophilus) microplus. The compounds tested were neem coated silver nanoparticles (N-Ag NPs), deltamethrin neem coated silver nanoparticles (DN-Ag NPs), 2, 3 dehydrosalannol (2,3 DHS), 2, 3 DHS coated silver nanoparticles (2, 3-DHS-Ag NPs), Quercetin dihydrate (QDH) and QDH coated silver nanoparticles (QDH-Ag NPs). Also included in this study, for the purpose of comparison, were neem leaf extract (NLE), silver nitrate (AgNO3) and deltamethrin (D). Acaricidal activity on larvae and adults of R. (B.) microplus was tested by larval packet test (LPT) and adult immersion test (AIT) respectively. In the LPT, 100% mortality was obtained at concentrations (ppm) of 360, 6000, 260, 200, 50, 300, 85, 600 and 200 for the compounds, D, NLE, Ag NO3, N-Ag NPs, DN-Ag NPs, 2, 3 DHS, 2, 3 DHS-Ag NPs, QDH, QDH-Ag NPs respectively. In AIT, the proportions of mortality and oviposition inhibition were proportionate but the reproductive index was inversely proportional to the concentration of the compounds used. The effect of DN-Ag NPs on mortality was the highest (93.33%) at 50ppm concentration. The mean reproductive index (0.01) and oviposition inhibition (99.16%) values were statistically significant when compared to control group. DN-Ag NPs showed significantly (P<0.05) lower LC50 (3.87ppm; 21.95ppm) and LC99 (53.05ppm; 90.06ppm) values against both the larvae and adults of R. (B.) microplus. The oviposition inhibiting ability of various compounds was determined to assess the reproductive performance of adult female ticks. The DN-Ag NPs had potent oviposition inhibitory activity with significantly lower IC50 and IC99 values compared to the rest of the treatments at 0.034 and 51.07ppm respectively. These results showed that the DN-Ag NPs had significant acaricidal activity against R. (B.) microplus.

  11. Invitro acaricidal activity of ethnoveterinary plants and green synthesis of zinc oxide nanoparticles against Rhipicephalus (Boophilus) microplus.

    PubMed

    Banumathi, Balan; Malaikozhundan, Balasubramanian; Vaseeharan, Baskaralingam

    2016-01-30

    The present study was designed to investigate the invitro acaricidal effects of seven ethnoveterinary plants, zinc acetate and green synthesized zinc oxide nanoparticles against the Southern cattle tick, Rhipicephalus (Boophilus) microplus. The selected ethnoveterinary plants were extracted using ethanol and aqueous (water) solvents at 0.02mg/ml and 0.04mg/ml concentrations. Of these seven plants, Lobelia leschenaultiana showed the highest percentage of tick mortality. The ethanol extracts of L. leschenaultiana showed 93.33% mortality at 0.04mg/ml and its LC50 was 0.05mg/ml. However, zinc acetate exhibited 70% mortality at 0.04mg/ml (LC50: 0.0192mg/ml). Further, we synthesized ZnO nanoparticle using the leaf extracts of L. leschenaultiana and zinc acetate as the precursor material to control R. (B.) microplus. The structural characterization of the synthesized ZnO nanoparticles (Ll-ZnO NPs) was performed by UV-vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM) and transmission electron microsopy (TEM). UV-vis spectra showed the absorption band at 383nm. XRD analysis clearly showed the crystalline nature of Ll-ZnO NPs with various Bragg's reflection peaks at 100, 002, 101, 102, 110, 103, 200, 201 and 202 planes. FTIR analysis showed the possible functional groups of Ll-ZnO NPs with strong band at 3420.63 and 2922.48cm(-1). SEM and TEM analysis revealed that the Ll-ZnO NPs were spherical and hexagonal in shape with particle size ranging between 20 and 65nm. The mortality of R. (B.) microplus after treatment with Ll-ZnO NPs was 35, 57.5 and 82.5% at 0.001, 0.002 and 0.004mg/ml. On the otherhand, 100% mortality of R. (B.) microplus was observed at 0.008mg/ml (LC50: 0.0017mg/ml). The results indicated that the Ll-ZnO NPs have good acaricidal properties compared to L. leschenaultiana leaf extract and zinc acetate.

  12. Comparative microarray analysis of Rhipicephalus (Boophilus) microplus expression profiles of larvae pre-attachment and feeding adult female stages on Bos indicus and Bos taurus cattle

    PubMed Central

    2010-01-01

    Background Rhipicephalus (Boophilus) microplus is an obligate blood feeder which is host specific to cattle. Existing knowledge pertaining to the host or host breed effects on tick transcript expression profiles during the tick - host interaction is poor. Results Global analysis of gene expression changes in whole R. microplus ticks during larval, pre-attachment and early adult stages feeding on Bos indicus and Bos taurus cattle were compared using gene expression microarray analysis. Among the 13,601 R. microplus transcripts from BmiGI Version 2 we identified 297 high and 17 low expressed transcripts that were significantly differentially expressed between R. microplus feeding on tick resistant cattle [Bos indicus (Brahman)] compared to R. microplus feeding on tick susceptible cattle [Bos taurus (Holstein-Friesian)] (p ≤ 0.001). These include genes encoding enzymes involved in primary metabolism, and genes related to stress, defence, cell wall modification, cellular signaling, receptor, and cuticle formation. Microarrays were validated by qRT-PCR analysis of selected transcripts using three housekeeping genes as normalization controls. Conclusion The analysis of all tick stages under survey suggested a coordinated regulation of defence proteins, proteases and protease inhibitors to achieve successful attachment and survival of R. microplus on different host breeds, particularly Bos indicus cattle. R. microplus ticks demonstrate different transcript expression patterns when they encounter tick resistant and susceptible breeds of cattle. In this study we provide the first transcriptome evidence demonstrating the influence of tick resistant and susceptible cattle breeds on transcript expression patterns and the molecular physiology of ticks during host attachment and feeding. The microarray data used in this analysis have been submitted to NCBI GEO database under accession number GSE20605 http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE20605. PMID:20637126

  13. Analysis of Babesia bovis-induced gene expression changes in the cattle tick, Rhipcephalus (Boophilus) microplus.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Boophilus ticks are vectors of Babesia bovis, the protozoan causative agent of cattle fever, a disease which is responsible for significant production losses to cattle producers in much of Africa, Central and South America and Australia. We utilized subtractive cDNA library synthesis techniques to o...

  14. Immunogenic potential of Rhipicephalus (Boophilus) microplus aquaporin 1 against Rhipicephalus sanguineus in domestic dogs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study evaluated a recombinant aquaporin 1 protein of Rhipicephalus microplus (RmAQP1) as antigen in a vaccine against R. sanguineus. Five dogs were vaccinated with RmAQP1 (10 µg) + adjuvant (Montanide) (G1), and five were inoculated with adjuvant only (G2), three times. Twenty-one days after th...

  15. CattleTickBase: An integrated Internet-based bioinformatics resource for Rhipicephalus (Boophilus) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Rhipicephalus microplus genome is large and complex in structure, making a genome sequence difficult to assemble and costly to resource the required bioinformatics. In light of this, a consortium of international collaborators was formed to pool resources to begin sequencing this genome. We have...

  16. Rhipicephalus (Boophilus) microplus aquaporin as an effective vaccine antigen to protect against cattle tick infestations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A cDNA encoding an aquaporin from the cattle tick, Rhipicephalus microplus, was isolated from transcriptomic studies. Bioinformatic analysis indicates this aquaporin, designated RmAQP1, shows greatest amino acid similarity to the human aquaporin 7 family. Members of this family of water-conducting c...

  17. Pharmacological characterization of a tyramine receptor from the southern cattle tick, Rhipicephalus (Boophilus) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The southern cattle fever tick (Rhipicephalus microplus) is a hematophagous external parasite that vectors the causative agents (Babesia spp.), which results in cattle fever or red water fever. The southern cattle fever tick is a threat to the cattle industry in many locations throughout the world. ...

  18. Rhipicephalus (Boophilus) microplus strain Deutsch, whole genome shotgun sequencing project first submission of genome sequence

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The size and repetitive nature of the Rhipicephalus microplus genome makes obtaining a full genome sequence difficult. Cot filtration/selection techniques were used to reduce the repetitive fraction of the tick genome and enrich for the fraction of DNA with gene-containing regions. The Cot-selected ...

  19. In vitro acaricidal activity of Murraya koenigii (L.) Spreng (Rutaceae) extracts against synthetic pyrethroid-resistant Rhipicephalus (Boophilus) microplus.

    PubMed

    Singh, Nirbhay Kumar; Jyoti; Vemu, Bhaskar; Singh, Harkirat; Prerna, Mranalini; Daundkar, Prashant S; Sharma, S K; Dumka, V K

    2015-04-01

    Larval packet test was used for detection of resistance status against cypermethrin and deltamethrin, the most commonly used synthetic pyrethroids in Rhipicephalus (Boophilus) microplus collected from Faridkot district, Punjab (India). The slope of mortality, lethal concentration for 50 % (LC50) and resistance levels were determined from the regression graphs of probit mortality of ticks plotted against log values of increasing concentrations of cypermethrin and deltamethrin. Results indicated presence of resistance of levels I and II against cypermethrin (resistance factor (RF) = 2.82) and deltamethrin (RF = 8.44), respectively. Adult immersion test was used to assess the acaricidal activity of aqueous (MLAq), ethanol (MLE), chloroform (MLC), acetone (MLA) and hexane (MLH) extracts of leaves of Murraya koenigii against these synthetic pyrethroid (SP)-resistant engorged adult females of R. (B.) microplus by determination of per cent adult mortality, reproductive index (RI), per cent inhibition of oviposition (%IO) and hatching rate. The per cent mortality caused by various extracts at concentrations ranging from 0.625 to 10.0% varied from 0.0 to 100.0% with maximum per cent mortality of 10.0, 100.0, 70.0, 40.0 and 10.0 recorded against MLAq, MLE, MLC, MLA and MLH, respectively. Among all extracts, the highest acaricidal property against SP-resistant R. (B.) microplus was exhibited by the MLE as it showed the minimum LC50 [95% confidence limit (CL)] values of 2.97% (2.82-3.12%), followed by MLC as 10.26% (8.84-11.91 %) and MLA as 18.22% (16.18-20.52%). The average egg mass weight recorded in live ticks treated with various concentrations of different extracts was lower than the respective control group ticks and was significantly (p < 0.01) lower in ticks treated with MLH extract. However, no significant effect on hatchability of eggs of treated groups when compared to control was recorded. A significant (p < 0.05) decrease in the RI was recorded in

  20. Acaricidal activity of the essential oil from Tetradenia riparia (Lamiaceae) on the cattle tick Rhipicephalus (Boophilus) microplus (Acari; Ixodidae).

    PubMed

    Gazim, Zilda Cristiani; Demarchi, Izabel Galhardo; Lonardoni, Maria Valdrinez Campana; Amorim, Ana Carolina L; Hovell, Ana Maria C; Rezende, Claudia Moraes; Ferreira, Gilberto Alves; de Lima, Edson Luiz; de Cosmo, Fábio Antunes; Cortez, Diogenes Aparício Garcia

    2011-10-01

    Tetradenia riparia (Lamiaceae) is a well-known herbal medicine with a variety of useful properties, including its acaricidal effect. This experiment was carried out to study the bioacaricidal activity of T. riparia essential oil (EO) against engorged females of Rhipicephalus (Boophilus) microplus (Acari; Ixodidae). For this purpose, nine serial concentrations (12.50%, 6.25%, 3.75%, 1.80%, 0.90%, 0.45%, 0.22%, 0.11%, and 0.056% w/v) of T. riparia were used for the adult immersion test (AIT). For the larval packet test (LPT), we used 14 serial concentrations (100.00%, 50.00%, 25.00%, 12.50%, 6.25%, 3.65%, 1.82%, 0.91%, 0.45%, 0.228%, 0.114%, 0.057%, 0.028%, and 0.014% w/v). The results for AIT showed 100.00% and 2.05% mortality, 19.00 and 90.20% for the total number of eggs, egg-laying inhibition of 0.00% and 90.20%, hatchability inhibition of 0.00% and 70.23%, and product effectiveness of 100.00% and 2.89%, respectively. The AIT indicated that the LC(50) and LC(99.9), calculated using the Probit test, were for mortality (%) 0.534g/mL (0.436-0.632) and 1.552g/mL (1.183-1.92); for total number of eggs were 0.449g/mL (0.339-0.558) and 1.76g/mL (1.27-2.248); and for hatchability inhibition were 0.114g/mL (0.0-0.31) and 2.462g/mL (1.501-3.422), respectively. Larvae between 14 and 21days old were fasted and placed in each envelope. Bioassays were performed at 27°±1°C, RH⩾80%. Larval mortality was observed 24h after treatment and showed 10.60-100% mortality in the LPT bioassay. The LPT showed that the LC(50) and LC(99.9) were 1.222g/mL (0.655-1.788) and 11.382g/mL (7.84-14.91), respectively. A positive correlation between T. riparia EO concentration and tick control, was observed by the strong acaricidal effects against R. (B.) microplus, and the mortality rate of ticks was dose-dependent. Our results showed that T. riparia is a promising candidate as an acaricide against resistant strains of R. (B.) microplus.

  1. In Vitro Acaricidal Activity of Atropa belladonna and Its Components, Scopolamine and Atropine, against Rhipicephalus (Boophilus) microplus

    PubMed Central

    Godara, R.; Katoch, M.; Yadav, Anish; Parveen, S.; Vij, Bhavna; Khajuria, Varun; Singh, G.; Singh, Nirbhay K.

    2014-01-01

    In vitro efficacy of methanolic extract of Atropa belladonna and its components scopolamine and atropine was assessed against Rhipicephalus (Boophilus) microplus. Five concentrations of the extract (1.25%, 2.5%, 5%, 10%, and 20%) were used whereas scopolamine and atropine were each tested at 0.1%. In adult immersion test, the extract was lethal to ticks at 20% concentration. The LC50 and LC95 values were determined as 6.875% and 17.306%, respectively. The extract caused a significant reduction (P < 0.05) in egg mass production at 10% concentration. In larval packet test, the extract was lethal to larvae in the concentrations of 10% and 20% after 24 h, with LC50 and LC95 values of 1.321% and 4.935%, respectively. Scopolamine and atropine showed 93.3% and 60.0% mortality of adult ticks, respectively, but they caused complete (100%) blocking of hatching as well as 100% larval mortality. Scopolamine and atropine were observed to be more potent than the crude extract at an equivalent concentration in both the bioassays. PMID:25516877

  2. Characterization of proteinases from the midgut of Rhipicephalus (Boophilus) microplus involved in the generation of antimicrobial peptides

    PubMed Central

    2010-01-01

    Background Hemoglobin is a rich source of biologically active peptides, some of which are potent antimicrobials (hemocidins). A few hemocidins have been purified from the midgut contents of ticks. Nonetheless, how antimicrobials are generated in the tick midgut and their role in immunity is still poorly understood. Here we report, for the first time, the contribution of two midgut proteinases to the generation of hemocidins. Results An aspartic proteinase, designated BmAP, was isolated from the midgut of Rhipicephalus (Boophilus) microplus using three chromatographic steps. Reverse transcription-quantitative polymerase chain reaction revealed that BmAP is restricted to the midgut. The other enzyme is a previously characterized midgut cathepsin L-like cysteine proteinase designated BmCL1. Substrate specificities of native BmAP and recombinant BmCL1 were mapped using a synthetic combinatorial peptide library and bovine hemoglobin. BmCL1 preferred substrates containing non-polar residues at P2 subsite and polar residues at P1, whereas BmAP hydrolysed substrates containing non-polar amino acids at P1 and P1'. Conclusions BmAP and BmCL1 generate hemocidins from hemoglobin alpha and beta chains in vitro. We postulate that hemocidins may be important for the control of tick pathogens and midgut flora. PMID:20663211

  3. Acaricidal activity of Lippia gracilis essential oil and its major constituents on the tick Rhipicephalus (Boophilus) microplus.

    PubMed

    Cruz, Elizangela Mércia de Oliveira; Costa, Livio Martins; Pinto, Jessika Andreza Oliveira; Santos, Darlisson de Alexandria; de Araujo, Sandra Alves; Arrigoni-Blank, Maria de Fátima; Bacci, Leandro; Alves, Péricles Barreto; Cavalcanti, Sócrates Cabral de Holanda; Blank, Arie Fitzgerald

    2013-07-01

    The present study aimed to evaluate the activity of Lippia gracilis Schauer essential oil obtained from different L. gracilis genotypes and their major components, carvacrol and thymol against Rhipicephalus (Boophilus) microplus (cattle tick) larvae and engorged females. The larval test was performed parallel to the adult immersion test for engorged females for four L. gracilis genotypes. Similar tests were further performed for their major compounds carvacrol and thymol. Carvacrol (LC50 of 0.22 and 4.46 mg/mL, to larvae and engorged females, respectively) was more efficient than thymol (LC50 of 3.86 and 5.50 mg/mL, to larvae and engorged females, respectively). The lethal concentrations obtained for the isolated essential oil from genotypes LGRA-201 against larvae (1.31 mg/mL) and LGRA-106 against engorged females (4.66 mg/mL) confirmed the acaricidal activity of L. gracilis essential oil and its effectiveness in controlling the southern cattle tick.

  4. Acaricidal activity of leaf extracts of Dalbergia sissoo Roxb. (Fabaceae) against synthetic pyrethroid resistant Rhipicephalus (Boophilus) microplus.

    PubMed

    Singh, Nirbhay Kumar; Jyoti; Vemu, Bhaskar; Prerna, Mranalini; Singh, Harkirat; Dumka, V K; Sharma, S K

    2016-06-01

    Resistance status of Rhipicephalus (Boophilus) microplus against synthetic pyrethroids was assessed by larval packet test which revealed level I and II resistance against cypermethrin and deltamethrin, respectively. Adult immersion test was employed to study the acaricidal activity of leaf extracts of Dalbergia sissoo (sheesham) against these ticks. Mortality and fecundity of ticks exposed to sheesham leaf aqueous (SLA) and ethanolic (SLE) extracts were evaluated at concentrations of 0.625, 1.25, 2.5, 5.0 and 10.0% and controls (distilled water and 10% ethanol). Higher acaricidal activity was recorded in SLA with a lower LC50 (95% CL) value of 1.58% (0.92-2.71%) than SLE [5.25% (4.91-5.63%)]. A significant decrease in egg mass weight and reproductive index was recorded in treated ticks along with an increase in percent inhibition of oviposition. A complete inhibition of hatching was recorded in eggs laid by ticks treated with higher concentrations of SLA, whereas, SLE exhibited no effect on hatching percentage.

  5. Simulation of control strategies for the cattle tick Boophilus microplus employing vaccination with a recombinant Bm86 antigen preparation.

    PubMed

    Labarta, V; Rodríguez, M; Penichet, M; Lleonart, R; Luaces, L L; de la Fuente, J

    1996-05-01

    Current strategies for the control of the cattle tick Boophilus microplus include the use of chemicals as the principal control method. These methods, however, have met with partially successful results. The recent development of immunological methods for the control of the cattle tick has opened new possibilities for the design of control strategies. Employing the results obtained by us in experiments testing the effect of vaccination with the recombinant vaccine, Gavac (Heber Biotec S.A.), on tick populations, we have developed a model to evaluate, through a computer program, the efficacy of the vaccine as a control method. The action of the vaccine on the control of tick populations was simulated and the specific serum antibody titers required to decrease the tick population in the field were calculated. The specific serum antibody titer required to decrease the tick population in the field after the first vaccination scheme was found to be > or = 57,200 and the antibody titer required to maintain this effect when the vaccine is already acting and after successive revaccinations was found to be > or = 27,500. Considerations about revaccination schemes and combination between vaccination and acaricide treatments as possible control strategies are discussed.

  6. Detection of amitraz and malathion resistance in field populations of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in Jammu region of India.

    PubMed

    Dutta, S; Godara, R; Katoch, R; Yadav, A; Katoch, M; Singh, N K

    2017-03-01

    Detection of resistance levels against amitraz and malathion in Rhipicephalus (Boophilus) microplus collected from four districts of Jammu region (India) was carried out using the adult immersion test. The regression graphs of probit mortality of ticks plotted against log values of concentration of drugs were utilised for the determination of slope of mortality, lethal concentration for 50% (LC50), 95% (LC95) and resistance factor (RF). On the basis of the data generated on variables (mortality, egg mass weight, reproductive index and percentage inhibition of oviposition) the resistance level was categorised as I, II, III and IV. Out of these four districts, resistance to amitraz was detected at level I in Udhampur (RF = 2.81), Jammu (RF = 2.53) and Samba isolates (RF = 2.24) whereas Rajouri isolate was found susceptible (RF = 1.0). Resistance to malathion was detected at level I in Udhampur (RF = 4.01) and Jammu isolates (RF = 1.76) whereas Rajouri (RF = 0.472) and Samba (RF = 0.199) isolates were found susceptible. The data generated on amitraz and malathion resistance status will help in formulating a tick control strategy in the region.

  7. Acaricidal activity of extracts from the leaves and aerial parts of Neoglaziovia variegata (Bromeliaceae) on the cattle tick Rhipicephalus (Boophilus) microplus.

    PubMed

    Dantas, A C S; Machado, D M R; Araujo, A C; Oliveira-Junior, R G; Lima-Saraiva, S R G; Ribeiro, L A A; Almeida, J R G S; Horta, M C

    2015-06-01

    This experiment was carried out to study the bioacaricidal activity of Neoglaziovia variegata against engorged females of Rhipicephalus (Boophilus) microplus. The mortality and fecundity of groups of engorged adult females exposed to different concentrations of ethanol, hexane, chloroform, and ethyl acetate extracts obtained from the leaves and aerial parts of N. variegata were evaluated, using three treatments with concentrations of 5, 10 e 25 mg/ml; two controls (distilled water and distilled water with drops of cremophor); with three replicates. The hexane extract of the leaves demonstrated significant results, presenting 94.1% inhibition of oviposition; 0.33% the average percentage of eclosion of eggs; and 99.8% of effectiveness. These results indicate N. variegata, particularly the hexane extract of leaves, as potential alternative control agents of R. (B.) microplus. Pharmacological and chemical studies are continuing in order to characterize the mechanism responsible for this effect.

  8. Effect of deforestation and introduction of exotic grasses as livestock forage on the population dynamics of the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in northern Argentina.

    PubMed

    Nava, Santiago; Mastropaolo, Mariano; Guglielmone, Alberto A; Mangold, Atilio J

    2013-12-01

    The effect of deforestation and the introduction of exotic grasses on the population dynamics of Rhipicephalus (Boophilus) microplus in northern Argentina was analysed. Biological parameters that were measured included proportion of females ovipositing, pre-oviposition period, incubation period of eggs, proportion of egg clusters hatching, larval longevity and total non-parasitic period. No significant differences were observed in proportion of females ovipositing and in pre-oviposition period between forested and grassland areas. Regarding the other parameters, in the majority of the temporal series there were no significant differences. In the cases where differences with statistical significance were detected, they were not unidirectional. The replacement of native forest by grasses can potentially increase tick abundance not by the modification of microclimatic conditions, but by increasing the tick-host encounter rate due to a higher cattle density. The hypothesis that deforestation and introduction of exotic grasses affects the non-parasitic phase of R. microplus in northern Argentina was not supported.

  9. Acaricidal effect and chemical composition of essential oils extracted from Cuminum cyminum, Pimenta dioica and Ocimum basilicum against the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Martinez-Velazquez, Moises; Castillo-Herrera, Gustavo Adolfo; Rosario-Cruz, Rodrigo; Flores-Fernandez, Jose Miguel; Lopez-Ramirez, Julisa; Hernandez-Gutierrez, Rodolfo; Lugo-Cervantes, Eugenia del Carmen

    2011-02-01

    Acaricidal activity of essential oils extracted from cumin seeds (Cuminum cyminum), allspice berries (Pimenta dioica) and basil leaves (Ocimum basilicum) were tested on 10-day-old Rhipicephalus (Boophilus) microplus tick larvae using the LPT. Two-fold dilutions of the three essential oils were tested from a starting dilution of 20% down to 1.25%. Results showed a high toxicological effect for cumin, producing 100% mortality in all tested concentrations on R. microplus larvae. Similarly, allspice essential oil produced 100% mortality at all concentrations with the exception of a dramatic decrease at 1.25% concentration. Conversely, basil essential oil was not shown to be toxic against R. microplus larvae. The most common compounds detected by gas chromatography-mass spectrometry were as follows: cumin: cuminaldehyde (22.03%), γ-terpinene (15.69%) and 2-caren-10-al (12.89%); allspice: methyl eugenol (62.7%) and eugenol (8.3%); basil: linalool (30.61%) and estragole (20.04%). Results clearly indicate that C. cyminum and P. dioica essential oils can be used as an effective alternative for R. microplus tick control, and there is a high probability they can be used for other ticks affecting cattle in Mexico and throughout the world, thereby reducing the necessity for traditional and unfriendly synthetic acaricides.

  10. Acaricidal activity of Cymbopogon winterianus, Vitex negundo and Withania somnifera against synthetic pyrethroid resistant Rhipicephalus (Boophilus) microplus.

    PubMed

    Singh, Nirbhay Kumar; Jyoti; Vemu, Bhaskar; Nandi, Abhijit; Singh, Harkirat; Kumar, Rajender; Dumka, V K

    2014-01-01

    Detection of resistance levels against cypermethrin and deltamethrin, the most commonly used synthetic pyrethroids (SPs) against Rhipicephalus (Boophilus) microplus collected from Moga, Punjab (India) was carried out using larval packet test. Results indicated the presence of resistance of level I and III against cypermethrin (resistance factors (RF) = 4.67) and deltamethrin (RF = 34.2), respectively. Adult immersion test was used to assess the acaricidal activity of aqueous and ethanolic extracts of leaves of Cymbopogon winterianus, Vitex negundo, and Withania somnifera along with roots of V. negundo against the SP resistant engorged females of R. (B.) microplus. The efficacy of various extracts was assessed by estimation of percent adult mortality, reproductive index (RI), percent inhibition of oviposition (%IO), and hatching rate. A concentration dependent increase in tick mortality was recorded which was more marked with various ethanolic extracts, and highest mortality was recorded in ticks treated with ethanolic extract of leaves of C. winterianus. The LC50 values were determined by applying regression equation analysis to the probit transformed data of mortality for various aqueous and ethanolic extracts. Acaricidal property was recorded to be higher in ethanolic extracts, and high activity was found with the ethanolic extract of leaves of C. winterianus with LC50 (95% CL) values of 0.46% (0.35-0.59%), followed by W. somnifera as 5.21% (4.45-6.09%) and V. negundo as 7.02% (4.58-10.74%). The egg mass weight of the live ticks treated with different concentrations of the various extract was significantly (p < 0.01) lower than that of control ticks; consequently, the RI and the %IO value of the treated ticks were reduced. Further, complete inhibition of hatching was recorded in eggs laid by ticks treated with ethanolic extracts of leaves of V. negundo and aqueous extracts of leaves of W. somnifera. The results of the current study indicate that extracts of C

  11. Inhibitor profile of bis(n)-tacrines and N-methylcarbamates on acetylcholinesterase from Rhipicephalus (Boophilus) microplus and Phlebotomus papatasi

    PubMed Central

    Swale, Daniel R.; Tong, Fan; Temeyer, Kevin B.; Li, Andrew; Lam, Polo C-H.; Totrov, Maxim M.; Carlier, Paul R.; Pérez de León, Adalberto A.; Bloomquist, Jeffrey R.

    2013-01-01

    The cattle tick, Rhipicephalus (Boophilus) microplus (Bm), and the sand fly, Phlebotomus papatasi (Pp), are disease vectors to cattle and humans, respectively. The purpose of this study was to characterize the inhibitor profile of acetylcholinesterases from Bm (BmAChE1) and Pp (PpAChE) compared to human and bovine AChE, in order to identify divergent pharmacology that might lead to selective inhibitors. Results indicate that BmAChE has low sensitivity (IC50 = 200 μM) toward tacrine, a monovalent catalytic site inhibitor with sub micromolar blocking potency in all previous species tested. Similarly, a series of bis(n)-tacrine dimer series, bivalent inhibitors and peripheral site AChE inhibitors possess poor potency toward BmAChE. Molecular homology models suggest the rBmAChE enzyme possesses a W384F orthologous substitution near the catalytic site, where the larger tryptophan side chain obstructs the access of larger ligands to the active site, but functional analysis of this mutation suggests it only partially explains the low sensitivity to tacrine. In addition, BmAChE1 and PpAChE have low nanomolar sensitivity to some experimental carbamate anticholinesterases originally designed for control of the malaria mosquito, Anopheles gambiae. One experimental compound, 2-((2-ethylbutyl)thio)phenyl methylcarbamate, possesses >300-fold selectivity for BmAChE1 and PpAChE over human AChE, and a mouse oral LD50 of >1500 mg/kg, thus providing an excellent new lead for vector control. PMID:24187393

  12. Acaricidal efficacies of Lippia gracilis essential oil and its phytochemicals against organophosphate-resistant and susceptible strains of Rhipicephalus (Boophilus) microplus.

    PubMed

    Costa-Júnior, Livio M; Miller, Robert J; Alves, Péricles B; Blank, Arie F; Li, Andrew Y; Pérez de León, Adalberto A

    2016-09-15

    Plant-derived natural products can serve as an alternative to synthetic compounds for control of ticks of veterinary and medical importance. Lippia gracilis is an aromatic plant that produces essential oil with high content of carvacrol and thymol monoterpenes. These monoterpenes have high acaricidal activity against Rhipicephalus (Boophilus) microplus. However, there are no studies that show efficacy differences of essential oils between susceptible and organophosphate resistant strains of R. (B.) microplus. The aim of the present study was to compare acaricidal effects of essential oils extracted from two different genotypes of L. gracilis and the main monoterpenes on larvae of both susceptible and organophosphate resistant R. (B.) microplus larvae. The efficacy of the essential oil of two genotypes of L. gracilis (106 and 201) and their monoterpenes carvacrol and thymol was measured using the larval immersion test on coumaphos-resistant and susceptible strains of R. (B.) microplus. Lethal concentrations were calculated using GraphPad Prism 6.0. Chemical analysis was performed by GC-MS and FID. Thymol and carvacrol were observed to be major constituents in 106 and 201L. gracilis genotype essential oils, respectively. Essential oils of both genotypes were more effective against organophosphate-resistant tick strain than susceptible tick strain. Carvacrol was 3.2 times more toxic to organophosphate resistant strain than to susceptible strain. Thymol was equally toxic to resistant and susceptible tick strains. The significantly higher efficacy monoterpene carvacrol against resistant ticks may lead to development of new natural product acaricide formulations for use to control organophosphate resistant R. (B.) microplus populations.

  13. Evaluation of stem aqueous extract and synthesized silver nanoparticles using Cissus quadrangularis against Hippobosca maculata and Rhipicephalus (Boophilus) microplus.

    PubMed

    Santhoshkumar, Thirunavukkarasu; Rahuman, Abdul Abdul; Bagavan, Asokan; Marimuthu, Sampath; Jayaseelan, Chidambaram; Kirthi, Arivarasan Vishnu; Kamaraj, Chinnaperumal; Rajakumar, Govindasamy; Zahir, Abdul Abduz; Elango, Gandhi; Velayutham, Kanayairam; Iyappan, Moorthy; Siva, Chinnadurai; Karthik, Loganathan; Rao, Kokati Venkata Bhaskara

    2012-10-01

    The present study was to determine the efficacies of anti-parasitic activities of synthesized silver nanoparticles (Ag NPs) using stem aqueous extract of Cissus quadrangularis against the adult of hematophagous fly, Hippobosca maculata (Diptera: Hippoboscidae), and the larvae of cattle tick, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae). Contact toxicity method was followed to determine the potential of parasitic activity. Twelve milliliters of stem aqueous extract of C. quadrangularis was treated with 88 ml of 1mM silver nitrate (AgNO(3)) solution at room temperature for 30 min and the resulting solution was yellow-brown color indicating the formation extracellular synthesis of Ag NPs. The synthesized Ag NPs were characterized with UV-visible spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Field emission scanning electron microscope (FESEM) and energy dispersive X-ray (EDX) spectroscopy. The synthesized Ag NPs were recorded by UV-visible spectrum at 420 nm and XRD patterns showed the nanoparticles crystalline in nature. FTIR analysis confirmed that the bioreduction of Ag((+)) ions to Ag NPs were due to the reduction by capping material of plant extract. FESEM image of Ag NPs showed spherical and oval in shape. By using the Bragg's Law and Scherrer's constant, the average mean size of synthesized Ag NPs was 42.46 nm. The spot EDX analysis showed the complete chemical composition of the synthesized Ag NPs. The mortality obtained by the synthesized Ag NPs from the C. quadrangularis was more effective than the aqueous extract of C. quadrangularis and AgNO(3) solution (1 mM). The adulticidal activity was observed in the aqueous extract, AgNO(3) solution and synthesized Ag NPs against the adult of H. maculata with LC(50) values of 37.08, 40.35 and 6.30 mg/L; LC(90) values of 175.46, 192.17 and 18.14 mg/L and r(2) values of 0.970, 0.992 and 0.969, respectively. The maximum efficacy showed in the aqueous extract, AgNO(3

  14. In vitro assessment of the acaricidal activity of computer-selected analogues of carvacrol and salicylic acid on Rhipicephalus (Boophilus) microplus.

    PubMed

    Concepción, Ramírez L; Froylán, Ibarra V; Herminia I, Pérez M; Norberto, Manjarrez A; Héctor J, Salgado Z; Yeniel, González C

    2013-10-01

    Rhipicephalus (Boophilus) microplus is a tick that causes huge economic losses in cattle. The indiscriminate use of acaricides has generated resistance to most compounds present on the market. This makes further investigation on other potential acaricides necessary, the in silico assay being an alternative to the design of new compounds. In the present study a biosilico assay was performed using TOMOCOMD-CARDD (TOpological MOlecular COMputer Design-Computer-Aided Rational Drug Design) and WEKA (Waikato Environment for Knowledge Analysis) software. Two carvacrol and four salicylic acid derivatives, synthesized by conventional methods and evaluated with the larval packet test on larvae of R. (B.) microplus were selected. All evaluated compounds presented acaricidal activity; however, ethyl 2-methoxybenzoate (91.8 ± 1.7 % mortality) and ethyl 2,5-dihydroxybenzoate (89.1 ± 1.6 % mortality) showed greater activity than salicylic acid. With regard to the carvacrol analogues, carvacrol acetate (67.8 ± 2.1 % mortality) and carvacrol methyl ether (71.7 ± 1.6 % mortality) also showed greater activity than carvacrol (35.9 ± 3.2 % mortality). TOMOCOMD-CARDD and WEKA software were helpful tools in the search for alternative structures with potential acaricidal activity on R. (B.) microplus.

  15. [Efficacy of extracts of plants in engorged females of Boophilus microplus from the mesoregion West of Maranhão, Brazil.].

    PubMed

    Costa, Francisco B; Vasconcelos, Pádua Suely Da S; Silva, Ana Maria M; Brandão, Vivian M; Da Silva, Iran A; Teixeira, Whaubtyfran C; Guerra, Rita De Maria S N; Dos Santos, Ana Clara G

    2008-09-01

    The objective was to evaluate the efficacy of hydro-alcoholic extracts of nim and citronela at 20% and eucalipto at 10% in Boophilus microplus engorged females collected in cattle naturally infected from the mesoregion West of Maranhão. At the laboratory the females were separated, weighted and distributed in six groups of 10, in duplicate. Each group was immersed in 10mL of the solution of the extracts, for two minutes. In the nim and citronela extracts there was 32% e 17%, respectively, while larval emergence the eucalipto extracts demonstrated 96% of efficacy. In the groups treated by Cipermetrina + Clorpirifós + Citronetal and Deltametrina (positive controls) the mortality occurred after 48h of treatment, while the groups immersed in distilled water (negative control) showed 100% of eggs mass and larval emergence. According to the results, it can be concluded that the extract of eucalipto could be used as acaricide in the control of B. microplus females since it was efficient in vitro, however to nim and citronela showed not efficacy. B. microplus females were not resistant to the chemical compounds used in this experiment.

  16. Molecular cloning and characterization of two novel autophagy-related genes belonging to the ATG8 family from the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Flores Fernández, José Miguel; Gutiérrez Ortega, Abel; Rosario Cruz, Rodrigo; Padilla Camberos, Eduardo; Alvarez, Angel H; Martínez Velázquez, Moisés

    2014-12-01

    Rhipicephalus (Boophilus) microplus is an obligate haematophagous arthropod and the major problem for cattle industry due to economic losses it causes. The parasite shows a remarkable adaptability to changing environmental conditions as well as an exceptional ability to survive long-term starvation. This ability has been related to a process of intracellular protein degradation called autophagy. This process in ticks is still poorly understood and only few autophagy-related (ATG) genes have been characterized. The aim of the present study was to examine the ESTs database, BmiGI, of R. microplus searching for ATG homologues. We predicted five putative ATG genes, ATG3, ATG4, ATG6 and two ATG8s. Further characterization led to the identification of RmATG8a and RmATG8b, homologues of GABARAP and MAP1LC3, respectively, and both of them belonging to the ATG8 family. PCR analyses showed that the expression level of RmATG8a and RmATG8b was higher in egg and larval stages when compared to ovary and midgut from adult ticks. This up-regulation coincides with the period in which ticks are in a starvation state, suggesting that autophagy is active in R. microplus.

  17. Reassociation kinetics-based approach for partial genome sequencing of the cattle tick, Rhipicephalus (Boophilus) microplus

    PubMed Central

    2010-01-01

    Background The size and repetitive nature of the Rhipicephalus microplus genome makes obtaining a full genome sequence fiscally and technically problematic. To selectively obtain gene-enriched regions of this tick's genome, Cot filtration was performed, and Cot-filtered DNA was sequenced via 454 FLX pyrosequencing. Results The sequenced Cot-filtered genomic DNA was assembled with an EST-based gene index of 14,586 unique entries where each EST served as a potential "seed" for scaffold formation. The new sequence assembly extended the lengths of 3,913 of the 14,586 gene index entries. Over half of the extensions corresponded to extensions of over 30 amino acids. To survey the repetitive elements in the tick genome, the complete sequences of five BAC clones were determined. Both Class I and II transposable elements were found. Comparison of the BAC and Cot filtration data indicates that Cot filtration was highly successful in filtering repetitive DNA out of the genomic DNA used in 454 sequencing. Conclusion Cot filtration is a very useful strategy to incorporate into genome sequencing projects on organisms with large genome sizes and which contain high percentages of repetitive, difficult to assemble, genomic DNA. Combining the Cot selection approach with 454 sequencing and assembly with a pre-existing EST database as seeds resulted in extensions of 27% of the members of the EST database. PMID:20540747

  18. Survey of acaricides resistance status of Rhipiciphalus (Boophilus) microplus collected from selected places of Bihar, an eastern state of India.

    PubMed

    Ghosh, Srikant; Kumar, Rinesh; Nagar, Gaurav; Kumar, Sachin; Sharma, Anil Kumar; Srivastava, Aman; Kumar, Suman; Ajith Kumar, K G; Saravanan, B C

    2015-07-01

    Monitoring acaricide resistance in field ticks and use of suitable managemental practices are essential for controlling tick populations infesting animals. In the present study, the acaricide resistance status in Rhipicephalus (Boophilus) microplus ticks infesting cattle and buffaloes of five districts located in the eastern Indian state, Bihar were characterized using three data sets (AIT, Biochemical assays and gene sequences). Adult immersion test (AIT) was adopted using seven field isolates and their resistance factor (RF) was determined. Six isolates (DNP, MUZ, BEG, VSH, DRB and SUL) were found resistant to both deltamethrin and diazinon and except VSH all were resistant to cypermethrin. One isolate (PTN) was susceptible with a RF below 1.5. To understand the possible mode of resistance development, targeted enzymes and gene sequences of the para sodium channel and achetylcholinesterase 2 (AChE2) were analyzed. The esterase, monooxygenase and glutathione-S-transferase (GST) activity of reference susceptible IVRI-I line was determined as 2.47 ± 0.007 nmol/min/mg protein, 0.089 ± 0.0016 nmol/mg of protein and 0.0439 ± 0.0003 nmol/mg/min respectively, which increased significantly in the resistant field isolates. However, except esterases, the fold increase of monooxygenase (1.14-2.27 times) and GST (0.82-1.53 times) activities were not very high. A cytosine (C) to adenine (A) nucleotide substitution (CTC to ATC) at position 190 in domain II S4-5 linker region was detected only in one isolate (SUL) having RF of 34.9 and in the reference deltamethrin resistant line (IVRI-IV). However, the T2134A mutation was not detected in domain IIIS6 transmembrane segment of resistant isolates and also in reference IVRI-IV line despite of varying degree of resistance. The flumethrin specific G215T and the recently identified T170C mutations were also absent in domain II sequences under study. Four novel amino acid substitutions in AChE2 gene of field isolates and in

  19. Phenotype changes inherited by crossing pyrethroid susceptible and resistant genotypes from the cattle tick Riphicephalus (Boophilus) microplus.

    PubMed

    Aguilar-Tipacamú, G; Rosario-Cruz, R; Miller, Robert J; Guerrero, Felix D; Rodriguez-Vivas, R I; García-Vázquez, Z

    2011-07-01

    Dialelic crosses and backcrosses of pyrethroid resistant (RR) and susceptible (SS) Rhipicephalus (Boophilus) microplus tick strains were carried out and the substitution (Phe-Ile) within the sodium channel gene was monitored in order to analyze the effects of the genotype on the pyrethroid resistance phenotype as measured by the larval packet test (LPT). Parental strains: susceptible (SS) and resistant (RR); dialelic crosses: RS (♂RR × ♀SS), and SR (♂SS × ♀RR); and backcrosses: RS × SS, RS × RR, SR × SS and SR × RR were infested on 280 kg calves. Resistance type (monogenic or polygenic) and effective dominance were determined based on the discriminant concentration (DC) for cipermethrine (0.5%), deltamethrine (0.09%) and flumethrine (0.01%). Allele specific PCR (AS-PCR) was used for genotyping, looking at a sodium channel mutation (Phe-Ile substitution). The mortality rates and allele frequency of susceptible and pyrethroid resistant reference strains were 0% mortality and 90% RR alleles for resistant strain, and 100% mortality and 0% RR alleles as measured by the larval packet test (LPT) and allele specific PCR (AS-PCR) respectively. Backcrossed strain SR × RR showed an effective dominance (D(ML)) of 0.605 for cypermethrin, 0.639 for deltamethrin and 0.498 for flumethrin, while survival of backcrosses RS × SS, RS × RR and SR × SS showed a significant tendency to recesivity. Backcrossed strain SR × RR (69.4%) also showed a higher RR genotype frequency with regards to RS × SS (25.5%), RS × RR (36.7%) and SR × SS (32.0%), however, susceptible allele was inherited in general as an incomplete dominant trait. Monogenic inheritance hypothesis was tested and the results showed monogenic inheritance for cypermethrin and flumethrin (P < 0.05) but not for deltamethrin (P > 0.05). However, significant correlation was found between RR genotype and the survival rate for all three pyrethroids used (P < 0.05), suggesting

  20. Acaricide rotation strategy for managing resistance in the tick Rhipicephalus (Boophilus) microplus (Acarina: Ixodidae): laboratory experiment with a field strain from Costa Rica.

    PubMed

    Thullner, Friederike; Willadsen, Peter; Kemp, David

    2007-09-01

    During the past two decades, resistance to pyrethroids within the cattle tick genus Boophilus has caused tick control problems in various tropical countries, mainly in Latin America, southern Africa, Australia, and New Caledonia. A Rhipicephalus (Boophilus) microplus (Canestrini) strain from Costa Rica, exhibiting resistance to the pyrethroid deltamethrin but only a very low resistance to organophosphates (OP) was kept under selection pressure for 9 to 11 generations by using deltamethrin or coumaphos (OP), either exclusively or in rotation. The objective of this acaricide rotation was to examine the possibility of delaying or reducing the full emergence of pyrethroid resistance. In the substrain selected with deltamethrin at the LD50 concentration, resistance to deltamethrin was measured after five generations (resistance factor [RF] = 9.2) and very high resistance after 11 generations (RF = 756), compared with the starting field strain from Costa Rica. In the substrain selected with deltamethrin then coumaphos in rotation, resistance to deltamethrin was variable from one generation to the next (RF = 1-4.2), but no high, stable resistance developed. After 10 generations of rotation, the deltamethrin RF was 1.6. In the substrains selected continuously with coumaphos or coumaphos and deltamethrin in rotation, no consistent change in resistance to coumaphos was observed. Rotation of deltamethrin with coumaphos seems to delay the development of strong resistance to deltamethrin in a population that had initially a low level of deltamethrin resistance.

  1. Two initial vaccinations with the Bm86-based Gavacplus vaccine against Rhipicephalus (Boophilus) microplus induce similar reproductive suppression to three initial vaccinations under production conditions

    PubMed Central

    2010-01-01

    Background The cattle tick, Rhipicephalus (Boophilus) microplus, affects livestock production in many regions of the world. Up to now, the widespread use of chemical acaricides has led to the selection of acaricide-resistant ticks and to environmental contamination. Gavacplus is a subunit vaccine based on the recombinant Bm86 tick antigen expressed in yeast, capable to control infestations of R. microplus under controlled and production conditions. The vaccine constitutes the core element of broad control programs against this ectoparasite, in which acquired immunity in cattle to Bm86 is combined with a rational use of acaricides. At present, the conventional vaccine scheme consists of three doses that should be administered at weeks 0, 4 and 7, followed by a booster every six months. Results In this study we assayed a reduction in the number of the initial doses of Gavacplus, evaluated the time course and the level of bovine anti-Bm86 antibodies elicited, and analyzed the vaccine effect on ticks engorging on immunized cattle under production conditions. Following three different immunization schemes, the bovines developed a strong and specific immune response characterized by elevated anti-Bm86 IgG titers. A reduction in the weight of engorging female ticks, in the weight of the eggs laid and also in R. microplus viable eggs percentage was obtained by using only two doses of Gavacplus administered at weeks 0 and 4, followed by a booster six months later. This reduction did not differ from the results obtained on ticks engorging on cattle immunized at weeks 0, 4 and 7. It was also demonstrated that anti-Bm86 antibody titers over 1:640, measured in bovines immunized at weeks 0 and 4, were sufficient to affect weight and reproductive potential of female ticks as compared with ticks engorging on unvaccinated animals. In addition, no statistically significant differences were detected in the average weight of eggs laid by ticks engorged on immunized cattle that showed

  2. Strategic applications of long-acting acaricides against Rhipicephalus (Boophilus) microplus in northwestern Argentina, with an analysis of tick distribution among cattle.

    PubMed

    Nava, Santiago; Mangold, Atilio J; Canevari, José T; Guglielmone, Alberto A

    2015-03-15

    Strategic applications of long-acting acaricides for the control of Rhipicephalus (Boophilus) microplus in northwestern Argentina were evaluated for one year. In addition, tick distribution among cattle was analyzed to evaluate if partial selective treatment or culling the small proportion of most heavily infested animals were feasible options to control R. (B.) microplus. Two different treatments schemes based on two applications of fluazuron and one application of 3.15% ivermectin were performed. Treatments were made in late winter and spring so as to act on the small 1st spring generation of R. (B.) microplus, in order to preclude the rise of the larger autumn generation. The overall treatment effect was positively significant in both schemes. The number of ticks observed in the control group was significantly higher than in the treated groups on all post-treatment counts. Group 2 exhibited more than 80% of efficacy almost throughout the study period, whereas Group 1 exhibited an efficacy percentage higher than 80% in September, October, December, February, April and May, but not in November (73.4%), January (58.3%), March (45.2%) or June (53.4%). Absolute control was observed in Group 2 in the counts of September and October, and in Group 1 in the count of February. The control strategies evaluated in this work provide an acceptable control level with only three applications of acaricides; at the same time, they prevent the occurrence of the autumn peak of tick burdens, which is characteristic of R. (B.) microplus in northwestern Argentina. Tick distribution was markedly aggregated in all counts. Although ticks were not distributed evenly among calves, the individual composition of the most heavily infested group was not consistent throughout the study period. In addition, the level of aggregation varied with tick abundance. These results suggest that applying acaricides to a portion of the herd or culling the most infested individuals at a given moment of the

  3. Molecular characterization and expression analysis of three novel autophagy-related genes from the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Flores Fernández, José Miguel; Barragán Álvarez, Carla Patricia; Sánchez Hernández, Carla Vanessa; Padilla Camberos, Eduardo; González Castillo, Celia; Ortuño Sahagún, Daniel; Martínez Velázquez, Moisés

    2016-11-01

    The cattle tick Rhipicephalus (Boophilus) microplus is a hematophagous ectoparasite of major importance for the livestock industry. It shows a remarkable ability to survive over long periods without feeding. However, the mechanisms used to endure long-term starvation are poorly understood. It is believed that autophagy, a process of intracellular protein degradation, may play a significant role to confront adverse environmental conditions. To advance our understanding of autophagy in R. microplus, in the present study we report the molecular characterization of three autophagy-related (ATG) genes, namely, RmATG3, RmATG4 and RmATG6, as well as their expression profiles in different developmental stages and organs of the parasite. The deduced amino acid sequences derived from the characterized gene sequences were subjected to Basic Local Alignment Search Tool analysis. The testing produced significant alignments with respective ATG proteins from Haemaphysalis longicornis and Ixodes scapularis ticks. Real-time polymerase chain reaction assays revealed that RmATG4 and RmATG6 transcripts were elevated in egg and ovary tissue, when compared with larva and midgut samples, while RmATG3 expression in midgut was 2-fold higher than in egg, larva and ovary samples.

  4. Resistance to cypermethrin, deltamethrin and chlorpyriphos in populations of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) from small farms of the State of São Paulo, Brazil.

    PubMed

    Mendes, M C; Lima, C K P; Nogueira, A H C; Yoshihara, E; Chiebao, D P; Gabriel, F H L; Ueno, T E H; Namindome, A; Klafke, G M

    2011-06-10

    A field survey of resistance was conducted based on the larval packet test technique with synthetic pyrethroids (cypermethrin and deltamethrin) and organophosphates (chlorpyriphos) in Rhipicephalus (Boophilus) microplus field populations from six different regions of the State of São Paulo (Brazil). 82.6% of the populations showed resistance to cypermethrin, 86.36% to deltamethrin and 65.25% to chlorpyriphos, with 50% presenting resistance to both SP and OP acaricide. According to the questionnaires completed by the producers, OP+SP mixtures followed by SP-only formulations were the products most commonly used for controlling the cattle tick in the surveyed areas. The present study showed high occurrence of resistance to SP and OP in the State of São Paulo, Brazil and revealed the type of strategy adopted by small dairy farms in this state. This information is fundamental in order to establish the monitoring of resistance on each farm individually, contributing to the rational use of acaricides for the control of R. (B.) microplus.

  5. [Determination of LC 90 and LT 90 of IBCB66 Beauveria bassiana (Ascomycetes: Clavicipitaceae) isolate for Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) control].

    PubMed

    Barci, Leila A G; de Almeida, José Eduardo M; de Campos Nogueira, Adriana H; do Prado, Angelo P

    2009-12-01

    The objective of this research was to evaluate the pathogenicity and the virulence of the IBCB66 isolate of Beauveria bassiana on infected larvae of Rhipicephalus (Boophilus) microplus. The IBCB66 fungus strain was used as standard isolates of B. bassiana against R. (B.) microplus larvae. The larval bioassay tests using the IBCB66 isolate were carried out to determine the (Lethal Concentration) LC50, LC90, (Lethal Time) LT50 and LT90. The IBCB66 fungus strain was tested at six different concentrations (5x10(6), 10(7), 5x10(7), 10(8), 5x10(8) and 10(9) conidia.mL(-1)) to determine the percentage of larval mortality. In addition, a Probit analysis was also performed. Total larval mortality was observed eighteen days after the beginning of the test in the group treated with 5x10(9) conidia.mL(1). The LC50 and LC90 were 3x10(7) and 5x10(8) respectively and the LT50 and LT90 were 10 and 16 days.

  6. Activation of several key components of the epidermal differentiation pathway in cattle following infestation with the cattle tick, Rhipicephalus (Boophilus) microplus.

    PubMed

    Kongsuwan, Kritaya; Josh, Peter; Colgrave, Michelle L; Bagnall, Neil H; Gough, Joanne; Burns, Brian; Pearson, Roger

    2010-03-15

    The cattle tick, Rhipicephalus (Boophilus) microplus, and the diseases it transmits pose a persistent threat to tropical beef production. Genetic selection of host resistance has become the method of choice for non-chemical control of cattle tick. Previous studies have suggested that larval stages are most susceptible to host resistance mechanisms. To gain insights into the molecular basis of host resistance that occurs during R. microplus attachment, we assessed the abundance of proteins (by isobaric tag for relative and absolute quantitation (iTRAQ) and Western blot analyses) and mRNAs (by quantitative reverse transcription PCR (qRT-PCR)) in skin adjacent to tick bite sites from high tick-resistant (HR) and low tick-resistant (LR) Belmont Red cattle following challenge with cattle tick. We showed substantially higher expression of the basal epidermal keratins KRT5 and KRT14, the lipid processing protein, lipocalin 9 (LCN9), the epidermal barrier catalysing enzyme transglutaminase 1 (TGM1), and the transcriptional regulator B lymphocyte-induced maturation protein 1 (Blimp1) in HR skin. Our data reveals the essential role of the epidermal permeability barrier in conferring greater resistance of cattle to tick infestation, and suggest that the physical structure of the epidermal layers of the skin may represent the first line of defence against ectoparasite invasion.

  7. Efficiency of sulphur in garlic extract and non-sulphur homeopathy in the control of the cattle tick Rhipicephalus (Boophilus) microplus.

    PubMed

    Costa, L M; Furlong, J

    2011-03-01

    The objective of the present work was to evaluate the efficacy of a non-sulphur-based homeopathic preparation and a sulphur-containing natural product derived from Allium sativum (Linnaeus) against infestation by the cattle tick Rhipicephalus (Boophilus) microplus (Canestrini) (Acari: Ixodidae). A total of 24 crossbred calves (7 : 8 Holstein : Zebu), aged 6-8 months and maintained in individual stables under tick-free conditions, were divided into three groups. Group 1 (control group) received no treatment; Group 2 was treated with 0.01 g/day of the homeopathic preparation Fator C&MC(®), and Group 3 was treated with 20 g/day of Enxofre-Allium sativum(®). After adaptation to the diet for 1 month, each calf was subjected to artificial infestation with 8000 R. (B.) microplus larvae (aged 7-14 days) twice per week over a 5-month period. Numbers of engorged females were recorded every 14 days and samples of freely released engorged females were collected at 14-day intervals commencing 3 months after the start of the experiment. The engorged females were weighed, incubated for 15 days under biochemical oxygen demand conditions at 27 ± 1 °C and relative humidity >85%, and the weights of the egg masses produced were recorded. Other biological parameters, including reproduction estimate, reduction in oviposition and efficiency of treatment, were determined. A significant reduction in the number of engorged females was detected on animals treated with Enxofre-Allium sativum(®) (Group 3) in comparison with the other two groups. The overall efficiency of the treatment with the sulphur-containing product was 64%, whereas that of the homeopathic preparation was 26%. Under the experimental conditions established, Enxofre-Allium sativum(®) can reduce the intensity of the R. (B.) microplus infestation.

  8. Soil quality influences efficacy of Melia azedarach (Sapindales: Meliaceae), fruit extracts against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Hexane extract of chinaberry, Melia azedarach L., unripe fruits obtained from different municipalities of Goias state in Brazil were evaluated on the southern cattle fever tick, Rhipicephalus microplus (Canestrini), engorged females. Hexanic extracts were assayed in decreasing concentrations from 0....

  9. Geostatistics and remote sensing using NOAA-AVHRR satellite imagery as predictive tools in tick distribution and habitat suitability estimations for Boophilus microplus (Acari: Ixodidae) in South America. National Oceanographic and Atmosphere Administration-Advanced Very High Resolution Radiometer.

    PubMed

    Estrada-Peña, A

    1999-02-01

    Remote sensing based on NOAA (National Oceanographic and Atmosphere Administration) satellite imagery was used, together with geostatistics (cokriging) to model the correlation between the temperature and vegetation variables and the distribution of the cattle tick, Boophilus microplus (Canestrini), in the Neotropical region. The results were used to map the B. microplus habitat suitability on a continental scale. A database of B. microplus capture localities was used, which was tabulated with the AVHRR (Advanced Very High Resolution Radiometer) images from the NOAA satellite series. They were obtained at 10 days intervals between 1983 and 1994, with an 8 km resolution. A cokriging system was generated to extrapolate the results. The data for habitat suitability obtained through two vegetation and four temperature variables were strongly correlated with the known distribution of B. microplus (sensitivity 0.91; specificity 0.88) and provide a good estimation of the tick habitat suitability. This model could be used as a guide to the correct interpretation of the distribution limits of B. microplus. It can be also used to prepare eradication campaigns or to make predictions about the effects of global change on the distribution of the parasite.

  10. In vitro and in vivo evaluation of cypermethrin, amitraz, and piperonyl butoxide mixtures for the control of resistant Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in the Mexican tropics.

    PubMed

    Rodriguez-Vivas, R I; Li, A Y; Ojeda-Chi, M M; Trinidad-Martinez, I; Rosado-Aguilar, J A; Miller, R J; Pérez de León, A A

    2013-10-18

    A study was conducted to evaluate the efficacy of cypermethrin, amitraz, and piperonyl butoxide (PBO) mixtures, through in vitro laboratory bioassays and in vivo on-animal efficacy trials, for the control of resistant Rhipicephalus (Boophilus) microplus on cattle in the Mexican tropics. Also, to examine mechanisms of resistance to cypermethrin in this tick population, the frequency of a mutated sodium channel gene (F1550I) was determined using a PCR assay. Results of laboratory bioassays using modified larval packet tests revealed that cypermethrin toxicity was synergized by PBO (from 46.6-57.0% to 83.7-85.0% larval mortality; P<0.05). The cypermethrin and amitraz mixture showed an additive effect (from 46.6-57.0% to 56.0-74.3% larval mortality). Strong synergism was observed with the mixture of cypermethrin+amitraz+PBO and this mixture was the most effective killing resistant tick larvae in vitro (96.7-100% of larval mortality). Tick larvae surviving exposure to cypermethrin or mixtures either with amitraz and PBO in vitro showed 2.9-49.6 higher probability to present the mutated allele than those killed by acaricide treatment (P<0.05). In the in vivo trial, the mixtures containing cypermethrin+PBO (80.6-97.3%), and cypermethrin+amitraz (87.0-89.7%) were more efficacious than cypermethrin alone (76.3-80.5%). The highest level of efficacy was obtained with the mixture of cypermethrin+amitraz+PBO, which yielded >95% control that persisted for 28 days post-treatment against R. microplus infesting cattle when tested under field conditions in the Mexican tropics. Although this mixture is a potentially useful tool to combat pyrethroid resistance, a product based on an acaricide mixture like the one tested in this study has to be used rationally.

  11. Integrated control of the southern cattle fever tick, Rhipicephalus (Boophilus) microplus, in Puerto Rico using new and alternative products

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Puerto Rico (PR) is plagued by the southern cattle fever tick (SCFT), Rhipicephalus microplus, which is considered the most economically important external parasite of livestock worldwide. A research coalition involving the livestock industry in PR, the PR Department of Agriculture (PR-DA), and the...

  12. Aquaporin 2 of Rhipicephalus (Boophilus) microplus as a potential target to control ticks and tick-borne parasites

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a collaboration with Washington State University and ARS-Pullman, WA researchers, we identified and sequenced a 1,059 base pair Rhipicephalus microplus transcript that contained the coding region for a water channel protein, Aquaporin 2 (RmAQP2). The clone sequencing resulted in the production of...

  13. New in vitro tests to evaluate the resistance level of the cattle tick, Rhipicephalus (Boophilus) microplus, against acaricides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of the present study was to compare two new in vitro tests designed to evaluate the resistance level of R. microplus with two tests currently recommended by the Food and Agriculture Organization of the United Nations (FAO): one performed on larvae, the Larval Packet Test (LPT) and one ...

  14. BmSI-7, a novel subtilisin inhibitor from Boophilus microplus, with activity toward Pr1 proteases from the fungus Metarhizium anisopliae.

    PubMed

    Sasaki, Sergio D; de Lima, Cássia A; Lovato, Diogo V; Juliano, Maria A; Torquato, Ricardo J S; Tanaka, Aparecida S

    2008-02-01

    BmSI-7 and BmSI-6, two Boophilus microplus subtilisin inhibitors (BmSI) were purified and characterized from eggs. The inhibitors isolated by classical purification methods presented molecular masses of 7408 and 7271Da, respectively, by MALDI-TOF-MS. Both BmSI-7 and BmSI-6 inhibited neutrophil elastase (K(i) 0.4 and 0.3nM) and subtilisin A (K(i) 1.4nM for both inhibitors). They also strongly inhibited Pr1 proteases from the fungus Metarhizium anisopliae; BmSI-7 (K(i) 50nM) and BmSI-6 (K(i) 2.2nM). The BmSI-7 full length cDNA was obtained using amino acid sequence information of BmSI-7 peptides generated by proteolytic digestion. BmSI-7 belongs to trypsin inhibitor like cysteine rich domain family (TIL), and it is transcribed in ovary, fat body, gut, salivary gland and haemocytes. BmSI-7 is the first TIL inhibitor described with inhibitory activity toward subtilisin A and Pr1 proteases of entomopathogenic fungi.

  15. A theoretical analysis of codon adaptation index of the Boophilus microplus bm86 gene directed to the optimization of a DNA vaccine.

    PubMed

    Ruiz, Lina María; Armengol, Gemma; Habeych, Edwin; Orduz, Sergio

    2006-04-21

    DNA vaccines utilize host cell molecules for gene transcription and translation to proteins, and the interspecific difference of codon usage is one of the major obstacles for effective induction of specific and strong immune response. In an attempt to improve codon usage effects of DNA vaccine on protein expression, a quantitative study was conducted to clarify the relationship of codon usage in the tick gene bm86 and its potential expression in bovine cells. The calculated relative synonymous codon usage (RSCU) and codon adaptation index (CAI) values of bm86 from Boophilus microplus and a set of 14 highly expressed genes from Bos taurus indicated that some codons utilized frequently in bm86 are rarely used in B. taurus genes and vice versa. The different translational efficiencies obtained suggested that after DNA vaccination using the wild bm86 gene, the protein Bm86 would be expressed in bovines, but it would not be the optimum sequence. However, using the codon-optimized bm86 gene to bovines, whose sequence was theoretically designed, would probably improve the level of the immune response generated against ticks.

  16. [Elaboration and evaluation of a candidate to the DNA vaccine using synthetic genes derived from the peptídeo SBm7462 against the carrapato Rhiphicephalus (Boophilus) microplus].

    PubMed

    Medeiros, Carla L; Mendonça, Bianca G; Tavares, Larissa C; Girão, Flávia A; Sossai, Sidimar; Peconick, Ana P; Carvalho, Gabriel D; Patarroyo, Joaquín H

    2008-09-01

    Rhiphicephalus (Boophilus) microplus is one of the most important arthropods in veterinary medicine due economic losses and health problems caused in cattle production. The vaccination represents optimum method evaluated with effective cost to prevent economic losses and to increase the duration and quality of life of the production animals. A synthetic peptide, SBm 7462, derived from Bm86, has been shown great results in control of ticks. The construction and synthesis of one nucleotide sequence based on this peptide might be useful for design a DNA vaccine that has many advances than peptide vaccine. A gene, called seq1, was constructed with a three repetition of nucleotide sequence of SBm 7462. It was cloned into a pCIneo vector expression in mammals and injected in BALB/c mouse. When mice were inoculated with the expression cassette they did not response in ELISA. They elevated antibody titles only when vaccinated with the synthetic peptide SBm7462®. And, the best titles of immunoglobulins were seen when the SBm7462® was administered subcutaneously.

  17. Identification and Structural-Functional Analysis of Cyclin-Dependent Kinases of the Cattle Tick Rhipicephalus (Boophilus) microplus

    PubMed Central

    Gomes, Helga; Romeiro, Nelilma C.; Braz, Gloria R. C.; de Oliveira, Eduardo Alves Gamosa; Rodrigues, Camilla; da Fonseca, Rodrigo Nunes; Githaka, Naftaly; Isezaki, Masayoshi; Konnai, Satoru; Ohashi, Kazuhiko; da Silva Vaz, Itabajara; Logullo, Carlos; Moraes, Jorge

    2013-01-01

    Cyclin-dependent kinases (CDKs) are a family of serine/threonine kinases essential for cell cycle progression. Herein, we describe the participation of CDKs in the physiology of Rhipicephalus microplus, the southern cattle tick and an important disease vector. Firstly, amino acid sequences homologous with CDKs of other organisms were identified from a R. microplus transcriptome database in silico. The analysis of the deduced amino acid sequences of CDK1 and CDK10 from R. microplus showed that both have caspase-3/7 cleavage motifs despite their differences in motif position and length of encoded proteins. CDK1 has two motifs (DKRGD and SAKDA) located opposite to the ATP binding site while CDK10 has only one motif (SLLDN) for caspase 3–7 near the ATP binding site. Roscovitine (Rosco), a purine derivative that inhibits CDK/cyclin complexes by binding to the catalytic domain of the CDK molecule at the ATP binding site, which prevents the transfer of ATP's γphosphoryl group to the substrate. To determine the effect of Rosco on tick CDKs, BME26 cells derived from R. microplus embryo cells were utilized in vitro inhibition assays. Cell viability decreased in the Rosco-treated groups after 24 hours of incubation in a concentration-dependent manner and this was observed up to 48 hours following incubation. To our knowledge, this is the first report on characterization of a cell cycle protein in arachnids, and the sensitivity of BME26 tick cell line to Rosco treatment suggests that CDKs are potential targets for novel drug design to control tick infestation. PMID:24146826

  18. Inorganic Polyphosphates Regulate Hexokinase Activity and Reactive Oxygen Species Generation in Mitochondria of Rhipicephalus (Boophilus) microplus Embryo

    PubMed Central

    Fraga, Amanda; Moraes, Jorge; da Silva, José Roberto; Costa, Evenilton P.; Menezes, Jackson; da Silva Vaz Jr, Itabajara; Logullo, Carlos; da Fonseca, Rodrigo Nunes; Campos, Eldo

    2013-01-01

    The physiological roles of polyphosphates (poly P) recently found in arthropod mitochondria remain obscure. Here, the possible involvement of poly P with reactive oxygen species generation in mitochondria of Rhipicephalus microplus embryos was investigated. Mitochondrial hexokinase and scavenger antioxidant enzymes, such as superoxide dismutase, catalase, and glutathione reductase were assayed during embryogenesis of R. microplus. The influence of poly P3 and poly P15 were analyzed during the period of higher enzymatic activity during embryogenesis. Both poly Ps inhibited hexokinase activity by up to 90% and, interestingly, the mitochondrial membrane exopolyphosphatase activity was stimulated by the hexokinase reaction product, glucose-6-phosphate. Poly P increased hydrogen peroxide generation in mitochondria in a situation where mitochondrial hexokinase is also active. The superoxide dismutase, catalase and glutathione reductase activities were higher during embryo cellularization, at the end of embryogenesis and during embryo segmentation, respectively. All of the enzymes were stimulated by poly P3. However, superoxide dismutase was not affected by poly P15, catalase activity was stimulated only at high concentrations and glutathione reductase was the only enzyme that was stimulated in the same way by both poly Ps. Altogether, our results indicate that inorganic polyphosphate and mitochondrial membrane exopolyphosphatase regulation can be correlated with the generation of reactive oxygen species in the mitochondria of R. microplus embryos. PMID:23983617

  19. [In Vitro efficacy of comercial formulations of ixocidides in engorged female of Boophilus microplus collected of dairy cattle at Paraíba Valey in the state of São Paulo].

    PubMed

    Pereira, José Roberto

    2006-01-01

    Laboratory trials were realized between the year 2001 at 2004 to evaluate the efficacy of commercial ixodicides against samples of Boophilus microplus collected of dairy bovines in 17 properties of eight cities at Paraíba Valey region, State of São Paulo Brazil. The trials was performed against engorged female using immersion technique. It was evidenced low efficacy of products of the chemical groups synthetic pyrethroid (28.24%) and amitraz (47.19%). The best efficacy was presented by products of the association between synthetic pyrethroid with organophosphorus (88.64%).

  20. Non-parasitic life cycle of the cattle tick Rhipicephalus (Boophilus) microplus in Panicum maximum pastures in northern Argentina.

    PubMed

    Mastropaolo, Mariano; Mangold, Atilio J; Guglielmone, Alberto A; Nava, Santiago

    2017-03-21

    The aim of this work was to study the non-parasitic phase of the Rhipicephalus microplus life cycle in Panicum maximum grasses from northern Argentina, in order to provide ecological information for designing methods of tick control. Four localities were chosen as replicates. The biological parameters measured were proportion of females ovipositing, the pre-oviposition period, the proportion of egg clusters hatching, the incubation period of eggs, larval longevity, and the total non-parasitic period (time from the exposure of the female to the date of death of the last larva) (TNPP)). The following general trends were observed: I) a longer TNPP occurred when female ticks were exposed in mid- and late summer and early spring; II) the shortest TNPP occurred when female ticks were exposed from late winter to late spring; III) larvae that were active in early and mid-summer had the shortest longevity; IV) incubation periods of eggs, which originated from females exposed in late summer, early autumn and mid-spring, were longer than the incubation period of eggs produced by females exposed in late spring and early summer; V) eggs did not hatch when the engorged females were exposed in the pastures in mid- and late autumn and winter. The spelling period of the P. maximum grasses that is needed to ensure total control of R. microplus consists of 19-20weeks if the spelling starts in late spring and early summer, and 27-28weeks if the spelling begins in mid- and late summer or in autumn.

  1. In vitro acaricidal properties of Semecarpus anacardium fruit and Datura stramonium leaf extracts against acaricide susceptible (IVRI-I line) and resistant (IVRI-V line) Rhipicephalus (Boophilus) microplus.

    PubMed

    Ghosh, Srikant; Tiwari, Shashi Shankar; Srivastava, Sharad; Kumar, Sachin; Sharma, Anil Kumar; Nagar, Gaurav; Kumar, K G Ajith; Kumar, Rajesh; Rawat, A K S

    2015-08-01

    In an attempt to identify plants having anti-tick properties, the 95% ethanolic and 50% hydro-ethanolic extracts of the fruits of Semecarpus anacardium and leaves of Datura stramonium were evaluated against reference tick lines of Rhipicephalus (Boophilus) microplus. The 95% ethanolic extracts of S. anacardium and D. stramonium caused 50% and 20% mortality, respectively, within 72 h of treatment by adult immersion test. The LC90 value of the ethanolic fruit extract of S. anacardium was determined as 13.5% (CI 12.05-15.12). The extract was also found efficacious (73.3%±3.3%) against the multi-acaricide-resistant IVRI-V line of R.(B.) microplus. The S. anacardium extract significantly affected the reproductive physiology of treated ticks by inhibiting the oviposition and was found safe. The HPTLC fingerprinting profile revealed the presence of pyrocatechol as a marker compound. The acaricidal property of S. anacardium against chemical acaricide-resistant R. (B.) microplus was discussed.

  2. Effect of vaccination with a recombinant Bm86 antigen preparation on natural infestations of Boophilus microplus in grazing dairy and beef pure and cross-bred cattle in Brazil.

    PubMed

    Rodríguez, M; Massard, C L; da Fonseca, A H; Ramos, N F; Machado, H; Labarta, V; de la Fuente, J

    1995-12-01

    Current methods for the control of the cattle tick Boophilus microplus infestations are not effective and the parasite remains a serious problem for the cattle industry in tropical and sub-tropical areas. Recent advances have introduced the possibility for the immunological control of the parasite through the use of recombinant vaccines. Recently, it was shown that the recombinant vaccine Gavac (Heber Biotec S.A.) is able to control B. microplus populations in artificially infected grazing dairy cattle in Cuba. To assay the effect of the vaccine on a different B. microplus strain and under different ecological conditions, we conducted a trial in Brazil on grazing dairy and beef pure and cross-bred cattle under natural infestation conditions. A farm in the northeast of the state of Sao Paulo was selected and two groups of animals per breed were included in the experiment and were maintained grazing on separate but similar pastures. For each breed, one group was vaccinated with the vaccine Gavac and the second group was not vaccinated and was employed as a control. In vaccinated cattle, during 36 weeks of experiment, the average infestation rate was maintained below 78 ticks per animal while average infestation peaks (mean +/- S.E.) of 144 +/- 44 ticks per animal (for dairy cross-bred cattle) and 195 +/- 42 ticks per animal (for beef cross-bred cattle) were recorded in the control groups. Tick infestation rates showed statistical significant differences (p = 0.04) between both experimental groups throughout the experiment. These results clearly showed, as in the Cuban study, that the vaccine controlled tick numbers in successive generations in the field.

  3. Population dynamics of the cattle tick Rhipicephalus (Boophilus) microplus in a subtropical subhumid region of Argentina for use in the design of control strategies.

    PubMed

    Canevari, J T; Mangold, A J; Guglielmone, A A; Nava, S

    2017-03-01

    The population dynamics of Rhipicephalus microplus (Ixodida: Ixodidae) in northwest Argentina was analysed to support the design of strategic methods for its control. Both parasitic and non-parasitic phases were studied. The seasonal activity of R. microplus in its parasitic phase was characterized by three peaks in abundance: the first in mid-late spring; the second in summer, and the third in autumn. The non-parasitic phase of R. microplus was characterized by a long total non-parasitic period observed after exposures of females from mid-summer to early autumn, a short total non-parasitic period observed after exposures of females from late winter to late spring, a short period of larval longevity in early and mid-summer, and no hatch of the eggs produced by females exposed in mid- and late autumn and winter. Treatments of cattle administered during the period from late winter to late spring will act on small cohorts of R. microplus, preventing the emergence of larger generations in summer and autumn. A 17-week spelling period starting in late spring and early summer will be necessary to achieve optimal control of R. microplus free-living larvae. If spelling begins in mid- or late summer or in autumn, the required period will be 26-27 weeks.

  4. In vitro activity of 3β-O-tigloylmelianol from Guarea kunthiana A. Juss (Meliaceae) on oogenesis and ecdysis of the cattle tick Rhipicephalus (Boophilus) microplus (Canestrini) (Acari: Ixodidae).

    PubMed

    Barbosa, Carolina da Silva; Borges, Lígia Miranda Ferreira; Louly, Carla Cristina Braz; Rocha, Thiago Lopes; de Sabóia-Morais, Simone Maria Teixeira; Miguita, Carlos Henrique; Garcez, Walmir Silva; Garcez, Fernanda Rodriguez

    2016-05-01

    We evaluated the effects of 3β-O-tigloylmelianol from Guarea kunthiana A. Juss (Meliaceae) on oogenesis, as a larvicide and on ecdysis of the larvae and the nymphs of the cattle tick Rhipicephalus (Boophilus) microplus (Canestrini) (Acari: Ixodidae). On the oogenesis' test, 48 engorged females were divided into three groups, evaluated at 24, 48 and 72 h post-treatment. Half of the females were treated with 0.01% 3β-O-tigloylmelianol diluted in distilled water and 5% dimethyl sulfoxide (DMSO), while the other half (controls) were exposed to distilled water and 5% DMSO. After treatment, the ovaries were weighed in order to measure the gonadosomatic index (GSI) and were also subjected to standard histological technical tests. On the larvicide and ecdysis' tests, 3β-O-tigloylmelianol was tested at concentrations of 0.01, 0.005, 0.0025 and 0.00125%. Compared with the controls, there was a reduction of GSI of approximately 50% on the treated group, which started at 48 h post treatment. Overall, the protolimonoid 3β-O-tigloylmelianol has caused a significant reduction in the number of oocytes. It has also caused alteration of the cytoplasmic and germinal vesicle diameters. Morphological changes, such as vacuolization, chorion irregularity which has modified the oocytes' morphology as well as alterations on the yolk's granules were also observed. The compound was not larvicide, however, interfered in the ecdysis of the larvae and the nymphs. This study shows that the protolimonoid 3β-O-tigloylmelianol from G. kunthiana acts on oogenesis and ecdysis of R. (B.) microplus, but not as larvicide, indicating that it acts on the endocrine system of the tick.

  5. Expressed gene sequences from adult ovary and adult female gut genes over-expressed upon Babesia bovis infection of Rhipcephalus (Boophilus) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Boophilus ticks are vectors of Babesia bovis, the protozoan causative agent of cattle fever, a disease which is responsible for significant production losses to cattle producers in much of Africa, Central and South America and Australia. We utilized subtractive cDNA library synthesis techniques to o...

  6. Simulated interactions of white-tailed deer (Odocoileus virginianus), climate variation and habitat heterogeneity on southern cattle tick (Rhipicephalus (Boophilus) microplus) eradication methods in South Texas, USA

    Technology Transfer Automated Retrieval System (TEKTRAN)

    White-tailed deer (odocoileus Virginiamis) is a host for cattle fever ticks, Rhipicephalus (Boophilus) sp.), which are vectors of the pathogens causing bovine babesiosis and anaplasmosis in cattle. Thus the potential role of white-tailed deer in compromising tick eradication efforts focused on cattl...

  7. Evolutionary conserved microRNAs are ubiquitously expressed compared to tick-specific miRNAs in the cattle tick Rhipicephalus (Boophilus) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    MicroRNAs (miRNAs) are small non-coding RNAs that act as regulators of gene expression in eukaryotes modulating a large diversity of biological processes. The discovery of miRNAs has provided new opportunities to understand the biology of a number of species. The cattle tick Rhipicephalus (Boophilus...

  8. In vitro and in vivo evaluation of deltamethrin and amitraz mixtures for the control of Rhipicephalus (Boophilus) microplus (Acari:Ixodidae) in New Caledonia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Deltamethrin and amitraz have been used to control R. microplus in New Caledonia for the past decade, and tick populations have developed resistance to both acaricides. A study was conducted to evaluate the effectiveness of deltamethrin and amitraz mixtures, through in vitro laboratory bioassays an...

  9. Molecular and pharmacological characterization of the first Chelicerata pyrokinin receptor from a worldwide tick vector of zoonotic pathogens, Rhipicephalus (Boophilus) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We identified the first pyrokinin receptor (Rhimi-PK-R) from the Chelicerata and analyzed structure-activity relationships of cognate ligand neuropeptides and their analogs. This receptor, which we cloned from larvae of the tick Rhipicephalus microplus (Acari: Ixodidae), is the ortholog of the inse...

  10. The expression of genes coding for distinct types of glycine-rich proteins varies according to the biology of three metastriate ticks, Rhipicephalus (Boophilus) microplus, Rhipicephalus sanguineus and Amblyomma cajennense

    PubMed Central

    2010-01-01

    Background Ticks secrete a cement cone composed of many salivary proteins, some of which are rich in the amino acid glycine in order to attach to their hosts' skin. Glycine-rich proteins (GRPs) are a large family of heterogeneous proteins that have different functions and features; noteworthy are their adhesive and tensile characteristics. These properties may be essential for successful attachment of the metastriate ticks to the host and the prolonged feeding necessary for engorgement. In this work, we analyzed Expressed Sequence Tags (ESTs) similar to GRPs from cDNA libraries constructed from salivary glands of adult female ticks representing three hard, metastriate species in order to verify if their expression correlated with biological differences such as the numbers of hosts ticks feed on during their parasitic life cycle, whether one (monoxenous parasite) or two or more (heteroxenous parasite), and the anatomy of their mouthparts, whether short (Brevirostrata) or long (Longirostrata). These ticks were the monoxenous Brevirostrata tick, Rhipicephalus (Boophilus) microplus, a heteroxenous Brevirostrata tick, Rhipicephalus sanguineus, and a heteroxenous Longirostrata tick, Amblyomma cajennense. To further investigate this relationship, we conducted phylogenetic analyses using sequences of GRPs from these ticks as well as from other species of Brevirostrata and Longirostrata ticks. Results cDNA libraries from salivary glands of the monoxenous tick, R. microplus, contained more contigs of glycine-rich proteins than the two representatives of heteroxenous ticks, R. sanguineus and A. cajennense (33 versus, respectively, 16 and 11). Transcripts of ESTs encoding GRPs were significantly more numerous in the salivary glands of the two Brevirostrata species when compared to the number of transcripts in the Longirostrata tick. The salivary gland libraries from Brevirostrata ticks contained numerous contigs significantly similar to silks of true spiders (17 and 8 in

  11. In vitro and in vivo acaricide action of juvenoid analogs produced from the chemical modification of Cymbopogon spp. and Corymbia citriodora essential oil on the cattle tick Rhipicephalus (Boophilus) microplus.

    PubMed

    Chagas, Ana Carolina S; Domingues, Luciana F; Fantatto, Rafaela R; Giglioti, Rodrigo; Oliveira, Márcia C S; Oliveira, Daniela H; Mano, Renata A; Jacob, Raquel G

    2014-09-15

    The present study aimed to evaluate the acaricidal action of the chemically modified essential oil of Cymbopogon spp. and Corymbia citriodora on Rhipicephalus (Boophilus) microplus. Citronellal was converted into N-butylcitronellylamine and in N-prop-2-inylcitronellylamine, analogs of juvenoids, by reductive amination using butylamine (N1 to N3) and propargylamine (N4 to N7). In vitro assays included the adult immersion, and larval packet tests. Engorged females were weighed in groups of 10 and tested in three replicates for six concentrations. They were immersed in the modified oils or control solution and incubated. In the larval packet test, the same substances and concentrations were evaluated in three replicates. In the in vivo test, six pastured heifers naturally infested with R. (B.) microplus were used per treatment: negative control, positive control (amitraz, Triatox(®)), original oil of C. citriodora at 1.5%, and modified oil containing 0.9% N-prop-2-inylcitronellylamine (N7). Ticks were counted in the right side of the body in 24 animals from day D-3 to D21. LC50 and LC90 were obtained by Probit analysis, while the in vivo results were log transformed and compared using the Tukey test. Among the nitrocellylamines tested in vitro, N6 was most effective on the engorged females (100% efficacy at 50mg/mL) and N7 on the larvae (100% efficacy at 6.25mg/mL). In the test with larvae, the original oil of C. citriodora was less effective than the counterpart modified oil (N7), proving that the chemical modification optimized its effect. In the in vivo test, no significant difference was observed between N7 and the negative control. The average numbers of ticks on the animals' right side were 32.8, 8.1, 37.9 and 35.4 for the negative control, positive control, original oil and N7, respectively. The chemical modification improved the efficacy in vitro, but it was not observed in vivo, perhaps due to the low stability of the amines under field conditions. The

  12. Rhipicephalus (Boophilus) microplus: Clotting time in tick-infested skin varies according to local inflammation and gene expression patterns in tick salivary glands

    PubMed Central

    Carvalho, Wanessa Araújo; Maruyama, Sandra Regina; Franzin, Alessandra Mara; Abatepaulo, Antônio Roberto Rodrigues; Anderson, Jennifer M.; Ferreira, Beatriz Rossetti; Ribeiro, José Marcos Chaves; Moré, Daniela Dantas; Maia, Antonio Augusto Mendes; Valenzuela, Jesus G.; Garcia, Gustavo Rocha; de Miranda Santos, Isabel K. Ferreira

    2010-01-01

    Ticks deposit saliva at the site of their attachment to a host in order to inhibit haemostasis, inflammation and innate and adaptive immune responses. The anti-haemostatic properties of tick saliva have been described by many studies, but few show that tick infestations or its anti-haemostatic components exert systemic effects in vivo. In the present study, we extended these observations and show that, compared with normal skin, bovine hosts that are genetically susceptible to tick infestations present an increase in the clotting time of blood collected from the immediate vicinity of haemorrhagic feeding pools in skin infested with different developmental stages of Rhipicepahlus microplus; conversely, we determined that clotting time of tick-infested skin from genetically resistant bovines was shorter than that of normal skin. Coagulation and inflammation have many components in common and we determined that in resistant bovines, eosinophils and basophils, which are known to contain tissue factor, are recruited in greater numbers to the inflammatory site of tick bites than in susceptible hosts. Finally, we correlated the observed differences in clotting times with the expression profiles of transcripts for putative anti-haemostatic proteins in different developmental stages of R. microplus fed on genetically susceptible and resistant hosts: we determined that transcripts coding for proteins similar to these molecules are overrepresented in salivary glands from nymphs and males fed on susceptible bovines. Our data indicate that ticks are able to modulate their host’s local haemostatic reactions. In the resistant phenotype, larger amounts of inflammatory cells are recruited and expression of anti-coagulant molecules is decreased tick salivary glands, features that can hamper the tick’s blood meal. PMID:20045690

  13. Acaricidal properties of vetiver essential oil from Chrysopogon zizanioides (Poaceae) against the tick species Amblyomma cajennense and Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Campos, Roseane Nunes de Santana; Nascimento Lima, Cecília Beatriz; Passos Oliveira, Alexandre; Albano Araújo, Ana Paula; Fitzgerald Blank, Arie; Barreto Alves, Péricles; Nascimento Lima, Rafaely; Albano Araújo, Vinícius; Santana, Alisson Silva; Bacci, Leandro

    2015-09-15

    Ticks are arthropods widely distributed in tropical and subtropical regions, which can transmit infectious agents also responsible for zoonoses. Excessive use of conventional acaricides has resulted in the onset of drug resistance by these parasites, thus the need to use alternative methods for their control. This study evaluated the acaricidal activities of Chrysopogon zizanioides (vetiver) essential oils containing different zizanoic and khuzimol (high and low acidity) acid concentrations on Amblyomma cajennense and Rhipicephalus microplus (Acari: Ixodidae). To this aims, toxicity tests of different concentrations of examined essential oils were conducted on adult females and larval stages. Results showed that the essential oils of C. zizanioides with high and low acidity reduced oviposition of females, eggs hatch and larval survival, being more effective than some commercial products widely used to control these ectoparasites. These results indicate that the C. zizanoides essential oils are promising candidates as acaricidal agents and represent also an add value to vetiver oil with high acidity, which is commercially undervalued in the cosmetic industry.

  14. Rhipicephalus microplus strain Deutsch, 10 BAC clone sequences

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cattle tick, Rhipicephalus (Boophilus) microplus, has a genome over 2.4 times the size of the human genome, and with over 70% of repetitive DNA, this genome would prove very costly to sequence at today's prices and difficult to assemble and analyze. We used labeled DNA probes from the coding reg...

  15. Rhipicephalus microplus strain Deutsch, whole genome shotgun sequencing project Version 2

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cattle tick, Rhipicephalus (Boophilus) microplus, has a genome over 2.4 times the size of the human genome, and with over 70% of repetitive DNA, this genome would prove very costly to sequence at today's prices and difficult to assemble and analyze. Cot filtration/selection techniques were used ...

  16. Reduced efficacy of commercial acaricides against populations of resistant cattle tick Rhipicephalus microplus from two municipalities of Antioquia, Colombia

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two distant Antioquian cattle farms where systemic and topical acaricides had previously failed to control infestations by Rhipicephalus (Boophilus) microplus were studied. An initial in vivo study was conducted using single subcutaneous injections with a long-acting formulation of ivermectin (630 µ...

  17. Prediction of G protein-coupled receptor encoding sequences from the synganglion transcriptome of the cattle tick, Rhipicephalus microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cattle tick, Rhipicephalus (Boophilus) microplus, is a pest which causes multiple health complications in cattle. The G-protein coupled receptor (GPCR) super-family presents an interesting target for developing novel tick control methods. However, GPCRs share limited sequence similarity among or...

  18. Exploring the use of anti-tick vaccine as tool for integrated eradication of the cattle fever tick, Rhipicephalus (Boophilus) annulatus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bovine babesiosis, also known as cattle fever, is a tick-borne protozoal disease foreign to the United States. It was eradicated by eliminating the vector species, Rhipicephalus (Boophilus) annulatus and R. (B.) microplus, through the efforts of the Cattle Fever Tick Eradication Program (CFTEP), wit...

  19. 9 CFR 72.1 - Ticks [Boophilus annulatus (Margaropus annulatus), Boophilus microplus, or Rhipicephalus evertsi...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Ticks ; interstate movement of... POULTRY) AND ANIMAL PRODUCTS TEXAS (SPLENETIC) FEVER IN CATTLE § 72.1 Ticks ; interstate movement of infested or exposed animals prohibited. No animals infested with ticks or exposed to tick infestation...

  20. 9 CFR 72.1 - Ticks [Boophilus annulatus (Margaropus annulatus), Boophilus microplus, or Rhipicephalus evertsi...

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Ticks ; interstate movement of... POULTRY) AND ANIMAL PRODUCTS TEXAS (SPLENETIC) FEVER IN CATTLE § 72.1 Ticks ; interstate movement of infested or exposed animals prohibited. No animals infested with ticks or exposed to tick infestation...

  1. 9 CFR 72.1 - Ticks [Boophilus annulatus (Margaropus annulatus), Boophilus microplus, or Rhipicephalus evertsi...

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Ticks ; interstate movement of... POULTRY) AND ANIMAL PRODUCTS TEXAS (SPLENETIC) FEVER IN CATTLE § 72.1 Ticks ; interstate movement of infested or exposed animals prohibited. No animals infested with ticks or exposed to tick infestation...

  2. 9 CFR 72.1 - Ticks [Boophilus annulatus (Margaropus annulatus), Boophilus microplus, or Rhipicephalus evertsi...

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Ticks ; interstate movement of... POULTRY) AND ANIMAL PRODUCTS TEXAS (SPLENETIC) FEVER IN CATTLE § 72.1 Ticks ; interstate movement of infested or exposed animals prohibited. No animals infested with ticks or exposed to tick infestation...

  3. Prediction of G protein-coupled receptor encoding sequences from the synganglion transcriptome of the cattle tick, Rhipicephalus microplus

    PubMed Central

    Guerrero, Felix D.; Kellogg, Anastasia; Ogrey, Alexandria N.; Heekin, Andrew M.; Barrero, Roberto; Bellgard, Matthew I.; Dowd, Scot E.; Leung, Ming-Ying

    2016-01-01

    The cattle tick, Rhipicephalus (Boophilus) microplus, is a pest which causes multiple health complications in cattle. The G protein-coupled receptor (GPCR) super-family presents a candidate target for developing novel tick control methods. However, GPCRs share limited sequence similarity among orthologous family members, and there is no reference genome available for R. microplus. This limits the effectiveness of alignment-dependent methods such as BLAST and Pfam for identifying GPCRs from R. microplus. However, GPCRs share a common structure consisting of seven transmembrane helices. We present an analysis of the R. microplus synganglion transcriptome using a combination of structurally-based and alignment-free methods which supplement the identification of GPCRs by sequence similarity. TMHMM predicts the number of transmembrane helices in a protein sequence. GPCRpred is a support vector machine-based method developed to predict and classify GPCRs using the dipeptide composition of a query aminoacid sequence. These two bioinformatic tools were applied to our transcriptome assembly of the cattle tick synganglion. Together, BLAST and Pfam identified 85 unique contigs as encoding partial or full length candidate cattle tick GPCRs. Collectively, TMHMM and GPCRpred identified 27 additional GPCR candidates that BLAST and Pfam missed. This demonstrates that the addition of structurally-based and alignment-free bioinformatic approaches to transcriptome annotation and analysis produces a greater collection of prospective GPCRs than an analysis based solely upon methodologies dependent upon sequence alignment and similarity. PMID:26922323

  4. Vaccination against ticks (Boophilus spp.): the experience with the Bm86-based vaccine Gavac.

    PubMed

    de la Fuente, J; Rodríguez, M; Montero, C; Redondo, M; García-García, J C; Méndez, L; Serrano, E; Valdés, M; Enríquez, A; Canales, M; Ramos, E; Boué, O; Machado, H; Lleonart, R

    1999-11-01

    The control of tick infestations and the transmission of tick-borne diseases remain a challenge for the cattle industry in tropical and subtropical areas of the world. Traditional control methods have been only partially successful and the parasites continue to result in significant losses for the cattle industry. Recently, vaccines containing the recombinant B. microplus gut antigen Bm86 have been developed. Our vaccine formulation (Gavac, Heber Biotec S.A., Havana, Cuba) has been registered and is commercially available in Cuba, Colombia, Dominican Republic, Brazil and Mexico. In controlled pen trials, Gavac has been effective for the control of artificial infestations of B. annulatus, B. decoloratus and chemical-sensitive and resistant B. microplus strains from Australia, Africa, America and Iran. In controlled field trials in Cuba, Brazil, Argentina and Mexico, Gavac has shown a 55-100% efficacy in the control of B. microplus infestations in grazing cattle 12-36 weeks after the first vaccination. Field trials under production conditions have been conducted in Cuba, Colombia, Brazil and Mexico in pure and cross-bred cattle herds. The application of Gavac has increased the time between acaricide treatments by an average of 32 /-21 days (P = 0.0005) resulting in important savings for the cattle industry. In Cuba, a cost-effectiveness analysis was conducted in more than 260000 animals. The cost-effectiveness analysis showed a 60% reduction in the number of acaricide treatments, together with the control of tick infestations and transmission of babesiosis, which resulted in savings of 23.4 dollars animal(-1) year (-1). These results clearly demonstrate the advantage of vaccination and support the application of Gavac for the control of Boophilus spp. infestations.

  5. Cloning and expression of a protective antigen from the cattle tick Boophilus microplus.

    PubMed Central

    Rand, K N; Moore, T; Sriskantha, A; Spring, K; Tellam, R; Willadsen, P; Cobon, G S

    1989-01-01

    Glycoproteins located on the luminal surface of the plasma membrane of tick gut epithelial cells, when used to vaccinate cattle, are capable of stimulating an immune response that protects cattle against subsequent tick infestation. One such tick gut glycoprotein, designated Bm86, has been purified to homogeneity and the amino acid sequences of peptide fragments generated by endoproteinase Lys-C digestion have been determined. We report here the isolation and characterization of a cDNA that encodes Bm86. The nucleotide sequence of the cDNA contains a 1982-base-pair open reading frame and predicts that Bm86 contains 650 amino acids including a 19-amino acid signal sequence and a 23-amino acid hydrophobic region adjacent to the carboxyl terminus. The main feature of the deduced protein sequence is the repeated pattern of 6 cysteine residues, suggesting the presence of several epidermal growth factor-like domains. A fusion protein consisting of 599 amino acids of Bm86 and 651 amino acids of beta-galactosidase was expressed in Escherichia coli as inclusion bodies. Ticks engorging on cattle vaccinated with these inclusion bodies were significantly damaged as a result of the immune response against the cloned antigen. Images PMID:2690068

  6. Acaricidal properties of Ricinus communis leaf extracts against organophosphate and pyrethroids resistant Rhipicephalus (Boophilus) microplus.

    PubMed

    Ghosh, Srikanta; Tiwari, Shashi Shankar; Srivastava, Sharad; Sharma, Anil Kumar; Kumar, Sachin; Ray, D D; Rawat, A K S

    2013-02-18

    Indian cattle ticks have developed resistance to commonly used acaricides and an attempt has been made to formulate an ecofriendly herbal preparation for the control of acaricide resistant ticks. A 95% ethanolic extract of Ricinus communis was used to test the efficacy against reference acaricide resistant lines by in vitro assay. In in vitro assay, the extract significantly affects the mortality rate of ticks in dose-dependent manner ranging from 35.0 ± 5.0 to 95.0 ± 5.0% with an additional effect on reproductive physiology of ticks by inhibiting 36.4-63.1% of oviposition. The leaf extract was found effective in killing 48.0, 56.7 and 60.0% diazinon, deltamethrin and multi-acaricide resistant ticks, respectively. However, the cidal and oviposition limiting properties of the extract were separated when the extract was fractionated with hexane, chloroform, n-butanol and water. The HPTLC finger printing profile of R. communis leaf extract under λ(max.) - 254 showed presence of quercetin, gallic acid, flavone and kaempferol which seemed to have synergistic acaricidal action. In vivo experiment resulted in 59.9% efficacy on Ist challenge, however, following 2nd challenge the efficacy was reduced to 48.5%. The results indicated that the 95% ethanolic leaf extract of R. communis can be used effectively in integrated format for the control of acaricide resistant ticks.

  7. Rhipicephalus (Boophilus) microplus resistant to acaricides and ivermectin in cattle farms of Mexico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ticks and the diseases they transmit cause great economic losses to livestock in tropical countries. Non-chemical control alternatives include the use of resistant cattle breeds, biological control, and vaccines. However, the most widely used method is the application of different chemical classes o...

  8. Development of a Spatially Targeted Field Sampling Technique for the Southern Cattle Tick, Rhipicephalus microplus, by Mapping Whitetailed Deer, Odocoileus virginianus, Habitat in South Texas

    PubMed Central

    Phillips, Pamela L.; Welch, John B.; Kramer, Matthew

    2014-01-01

    The objective of our study was to determine whether satellite remote sensed data could be used to identify white-tailed deer, Odocoileus virginianus (Zimmerman) (Artiodactyla: Cervidae), habitat and target locations for sampling free-living larvae of the southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodida: Ixodidae) in South Texas. Two methods for mapping white-tailed deer habitat were used, an object-oriented method to identify closed canopies and waterways for deer movement and two vegetation indices: the Normalized Difference Vegetation Index and the Modified Soil Adjusted Vegetation Index to identify forage for deer. These two data sets of favorable white-tailed deer habitat were combined within a geographic information system to identify locations for sampling ticks. Larvae of R. (B.) microplus, were sampled in Zapata County, Texas, by walking transects with attached flannel panels to jeans. Although the data set and sampling period were limited, data analysis demonstrated that sampling of free-living larvae of R. (B.) microplus can be conducted in South Texas, and larvae were most abundant in areas that harbored O. virginianus. Spatial analysis of satellite imagery to classify white-tailed deer/southern cattle tick habitat proved efficacious and may be useful in directing sampling activities in the field. PMID:25368044

  9. Development of a spatially targeted field sampling technique for the southern cattle tick, Rhipicephalus microplus, by mapping white-tailed deer, Odocoileus virginianus, habitat in South Texas.

    PubMed

    Phillips, Pamela L; Welch, John B; Kramer, Matthew

    2014-01-01

    The objective of our study was to determine whether satellite remote sensed data could be used to identify white-tailed deer, Odocoileus virginianus (Zimmerman) (Artiodactyla: Cervidae), habitat and target locations for sampling free-living larvae of the southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodida: Ixodidae) in South Texas. Two methods for mapping white-tailed deer habitat were used, an object-oriented method to identify closed canopies and waterways for deer movement and two vegetation indices: the Normalized Difference Vegetation Index and the Modified Soil Adjusted Vegetation Index to identify forage for deer. These two data sets of favorable white-tailed deer habitat were combined within a geographic information system to identify locations for sampling ticks. Larvae of R. (B.) microplus, were sampled in Zapata County, Texas, by walking transects with attached flannel panels to jeans. Although the data set and sampling period were limited, data analysis demonstrated that sampling of free-living larvae of R. (B.) microplus can be conducted in South Texas, and larvae were most abundant in areas that harbored O. virginianus. Spatial analysis of satellite imagery to classify white-tailed deer/southern cattle tick habitat proved efficacious and may be useful in directing sampling activities in the field.

  10. Laboratory evaluation of verbutin as a synergist of acaricides against larvae of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Synergistic effects of verbutin, a member of aryl alkynyl derivatives, to three commonly used acaricides were evaluated with the modified Food and Agricultural Organization Larval Packet Test (FAO-LPT) against both susceptible and resistant strains of the southern cattle tick Rhipicephalus (Boophilu...

  11. Inhibitor Profile of bis(n)-tacrines and N-methylcarbamates on Acetylcholinesterase from Rhipicephalus (Boophilus) Microplus and Phlebotomus Papatasi

    DTIC Science & Technology

    2013-03-28

    against rBmAChE (Table 2), but is 43-fold less effective against rPpAChE (Table 2). Commercial carbamate insecticides ( propoxur , carbaryl , and...85–92 87 the most active carbamate was carbofuran, whereas in the tick propoxur was the least active, and for the sandfly it was carbaryl . Although...populations. 2. Methods 2.1. Inhibitors, solvents, and assay reagents Propoxur (purity P 99%), bendiocarb (purity P 99%), edropho- nium (purity P 98

  12. Association of BoLA-DRB3.2 alleles with tick (Boophilus microplus) resistance in cattle.

    PubMed

    Martinez, M L; Machado, M A; Nascimento, C S; Silva, M V G B; Teodoro, R L; Furlong, J; Prata, M C A; Campos, A L; Guimarães, M F M; Azevedo, A L S; Pires, M F A; Verneque, R S

    2006-08-31

    Losses caused by bovine tick burdens in tropical countries have a tremendous economic impact on production systems. Besides reducing production, this parasite can cause death in the most susceptible animals. The use of commercial acaricides has been the major method of control, but their misuse has led to tick resistance to many chemicals. More recently, vaccines have been used in some countries without solving the problem completely. An alternative could be the development of resistant animals and the use of genetic markers and candidate genes that could help with the enormous task of selecting resistant animals. The bovine lymphocyte antigen genes (BoLA) have been shown to be associated with some parasitic infestations and disease incidence. Thus, the objective of the present study was to determine the association of BoLA-DRB3.2 alleles with tick resistance in cattle. The study was conducted on 231 F2 (Gyr x Holstein) animals that were artificially infested with 10,000 tick larvae. Log of tick count +1 was used as the dependent variable in a mixed animal model with allele substitution effects in addition to fixed effects of year and season at tick count, sex of calves, age of animal at tick count, hair type (short-straight, short-curl, long-straight, and long-curl), coat color (white, >75% white, 50- 75% white, and 25-50% white), and additive genetic, permanent environmental and residual effects as random. Females showed fewer ticks than males. Animals with short-straight hair were more resistant to tick infestation than animals with long-curl hair, and animals with whiter coat color also had fewer ticks. An association between BoLA alleles and lower tick number was found for alleles DRB3.2 *18, *20 and *27 at the 5% significance level. Also, one allele (DRB3.2*16) showed an association at the 10% level. Allele *27 was the most frequent in the population (30.7%), followed by alleles *16 (10.8%), *20 (8.7%) and *18 (2.4%). These results suggest that BoLA-DRB3.2 alleles could be used to help in the selection of animals resistant to tick infestation. However, further studies involving a larger population of cattle in combination with other BoLA genes may help to understand the mechanisms of resistance to parasites.

  13. Acetylcholinesterase of Rhipicephalus (Boophilus) microplus and Phlebotomus papatasi: Gene Identification, Expression, and Biochemical Properties of Recombinant Proteins

    DTIC Science & Technology

    2013-01-01

    Bushland U.S. Livestock Insects Research Laboratory, United States Department of Agricultural-Agricultural Research Service2, 2700 Fredericksburg Road...stranded RNA ; OP, organophosphate; OP-R, organophosphate resistant; OP-S, organophosphate susceptible; PCR, polymerase chain reaction; PpAChE, Phlebotomus... RNA , ribonucleic acid; RNAi, RNA interference (gene silencing). q Disclaimer: The views expressed in this article are those of the authors and do

  14. Suppressive subtractive hybridization analysis of Rhipicephalus (Boophilus) microplus larval and adult transcript expression during attachment and feeding

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ticks, as blood-feeding ectoparasites, affect their hosts both directly and as vectors of viral, bacterial and protozoal diseases. The tick’s mode of feeding means it must maintain intimate contact with the host in the face of host defensive responses for a prolonged time. The parasite: host interac...

  15. Reduced Efficacy of Commercial Acaricides Against Populations of Resistant Cattle Tick Rhipicephalus microplus from Two Municipalities of Antioquia, Colombia.

    PubMed

    Lopez-Arias, Anderson; Villar-Argaiz, David; Chaparro-Gutierrez, Jenny J; Miller, Robert J; Perez de Leon, Adalberto A

    2014-01-01

    Two distant Antioquian cattle farms where systemic and topical acaricides had previously failed to control infestations by Rhipicephalus (Boophilus) microplus were studied. An initial in vivo study was conducted using single subcutaneous injections with a long-acting formulation of ivermectin (630 μg/kg). Injections were made at 3-month intervals on animals at each farm to evaluate the therapeutic and persistent efficacy of ivermectin against R. microplus. Body tick counts and reproductive parameters of semi- or fully engorged females (≥5 mm) were assessed at 10-day intervals, and since no negative control group could be included, values were compared against those for day 0. Although there was an overall reduction of 50%-75% in tick numbers that persisted for 30-40 days, it was not significantly different at one of the farms and not enough to afford protection from severe infestations. The engorgement weight and egg mass weight of ticks from treated animals were significantly lower throughout the 50-day posttreatment period. Egg hatch was not significantly reduced posttreatment and remained at levels of 80%-90%. A random selection of 9 out of 28 commercial formulations of ivermectin sold in Colombia were analyzed by High Performance Liquid Chromatography (HPLC). All were within the expected labeled concentration (±15% deviation) of 1% and 3.15% ivermectin except for one. A popular unregistered injectable widely used in both farms and labeled as "natural pyrethrin", was found to contain 10.5% ivermectin. An adult immersion test was conducted to evaluate the efficacy of topical acaricides to recommended concentrations of five commercial products and/or their combinations. Efficacy was determined by comparing the reproductive index of each treated group to that of the control group. Cypermethrin (150 ppm) was completely ineffective at both farms. Amitraz (208 ppm) exhibited low and intermediate efficacies of 14% and 56%. The combination of amitraz (100 ppm) and

  16. Reduced Efficacy of Commercial Acaricides Against Populations of Resistant Cattle Tick Rhipicephalus microplus from Two Municipalities of Antioquia, Colombia

    PubMed Central

    Lopez-Arias, Anderson; Villar-Argaiz, David; Chaparro-Gutierrez, Jenny J; Miller, Robert J; Perez de Leon, Adalberto A

    2014-01-01

    Two distant Antioquian cattle farms where systemic and topical acaricides had previously failed to control infestations by Rhipicephalus (Boophilus) microplus were studied. An initial in vivo study was conducted using single subcutaneous injections with a long-acting formulation of ivermectin (630 μg/kg). Injections were made at 3-month intervals on animals at each farm to evaluate the therapeutic and persistent efficacy of ivermectin against R. microplus. Body tick counts and reproductive parameters of semi- or fully engorged females (≥5 mm) were assessed at 10-day intervals, and since no negative control group could be included, values were compared against those for day 0. Although there was an overall reduction of 50%–75% in tick numbers that persisted for 30–40 days, it was not significantly different at one of the farms and not enough to afford protection from severe infestations. The engorgement weight and egg mass weight of ticks from treated animals were significantly lower throughout the 50-day posttreatment period. Egg hatch was not significantly reduced posttreatment and remained at levels of 80%–90%. A random selection of 9 out of 28 commercial formulations of ivermectin sold in Colombia were analyzed by High Performance Liquid Chromatography (HPLC). All were within the expected labeled concentration (±15% deviation) of 1% and 3.15% ivermectin except for one. A popular unregistered injectable widely used in both farms and labeled as “natural pyrethrin”, was found to contain 10.5% ivermectin. An adult immersion test was conducted to evaluate the efficacy of topical acaricides to recommended concentrations of five commercial products and/or their combinations. Efficacy was determined by comparing the reproductive index of each treated group to that of the control group. Cypermethrin (150 ppm) was completely ineffective at both farms. Amitraz (208 ppm) exhibited low and intermediate efficacies of 14% and 56%. The combination of amitraz (100

  17. A GPCR-focused investigation of the R. microplus transcriptome

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rhipicephalus microplus, also known as the southern cattle tick, has been found in tropical and subtropical regions all over the world, including Mexico. It is a vector for parasites responsible for cattle diseases that can lead to decreased weight, anemia, loss of milk/meat production, and death. T...

  18. Generation of Full-Length cDNAs for Eight Putative GPCnR from the Cattle Tick, R. microplus Using a Targeted Degenerate PCR and Sequencing Strategy

    PubMed Central

    Corley, Sean W.; Piper, Emily K.; Jonsson, Nicholas N.

    2012-01-01

    We describe here a rapid and efficient method for the targeted isolation of specific members of gene families without the need for cloning. Using this strategy we isolated full length cDNAs for eight putative G-protein coupled neurotransmitter receptors (GPCnR) from the cattle tick Rhipicephalus (Boophilus) microplus. Gene specific degenerate primers were designed using aligned amino acid sequences of similar receptor types from several insect and arachnid species. These primers were used to amplify and sequence a section of the target gene. Rapid amplification of cDNA ends (RACE) PCR was used to generate full length cDNA sequences. Phylogenetic analysis placed 7 of these sequences into Class A G-protein coupled receptors (GPCR) (Rm_α2AOR, Rm_β2AOR, Rm_Dop1R, Rm_Dop2R, Rm_INDR, Rm_5-HT7R and Rm_mAchR), and one into Class C GPCR (Rm_GABABR). Of the 7 Class A sequences, only Rm_mAchR is not a member of the biogenic amine receptor family. The isolation of these putative receptor sequences provides an opportunity to gain an understanding of acaricide resistance mechanisms such as amitraz resistance and might suggest possibilities for the development of new acaricides. PMID:22403662

  19. Nomuraea rileyi as biological control agents of Rhipicephalus microplus tick.

    PubMed

    Perinotto, W M S; Terra, A L M; Angelo, I C; Fernandes, É K K; Golo, P S; Camargo, M G; Bittencourt, V R E P

    2012-10-01

    Nomuraea rileyi, a fungus pathogenic to insects, has been widely used for biological control of agricultural pests in Brazil. This study investigates the effects of N. rileyi, isolates Nr 138, Nr 151, and Nr 177, to eggs, larvae, and engorged females of Rhipicephalus microplus tick. Specimens were immersed in 1 ml of conidial suspension for 3 min, whereas the control group was immersed in 0.01% Tween 80 water solution. The isolate Nr 138 controlled 67.37% of ticks when the highest conidial concentration was used, 10(8) conidia ml(-1). The isolate Nr 177 significantly reduced the percentage of hatch of larvae from eggs treated with 10(8) conidia ml(-1). Conversely, the isolate Nr 151 was not virulent to eggs, larvae, or adults. Variability in virulence was observed among the N. rileyi isolates investigated in the current study-Nr 138 was more virulent to engorged females, while Nr 177 was more virulent to unfed larvae. Although N. rileyi proved to be virulent to several stages of R. microplus, the results obtained in this study indicate that N. rileyi does not appear to be a remarkable biological control agent for R. microplus.

  20. Acaricide activity in vitro of Acmella oleracea against Rhipicephalus microplus.

    PubMed

    Castro, K N C; Lima, D F; Vasconcelos, L C; Leite, J R S A; Santos, R C; Paz Neto, A A; Costa-Júnior, L M

    2014-10-01

    Cattle tick control has been limited by the resistance of these parasites to synthetic acaricides. Natural products are a possible alternative as they have different mechanisms of action. Acmella oleracea is a native plant with a large cultivated area in the Amazon region and could be easily used for large-scale preparation of a commercial product. This study evaluated the in vitro action of the hexane extract of the aerial parts of A. oleracea on larvae and engorged females of the cattle tick Rhipicephalus microplus. Spilanthol was the major constituent with a content of 14.8% in the extract. The hexane extract of A. oleracea was highly effective against larvae of R. microplus with an LC50 of 0.8 mg mL(-1). Against engorged females, hexane extract of A. oleracea reduced oviposition and hatchability of eggs with an LC50 of 79.7 mg mL(-1). Larvae and engorged females were killed by the hexane extract with high efficiency (>95%) at concentrations of 3.1 and 150.0 mg mL(-1), respectively. These results demonstrate that the hexane extract of A. oleracea has significant activity against R. microplus and has potential to be developed into formulations for tick control.

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

    PubMed

    Rodríguez-Vivas, R I; Ojeda-Chi, M M; Rosado-Aguilar, J A; Trinidad-Martínez, I C; Torres-Acosta, J F J; Ticante-Perez, V; Castro-Marín, J M; Tapia-Moo, C A; Vázquez-Gómez, G

    2013-08-01

    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.

  2. Toxic effects of various solvents against Rhipicephalus (Boophilus) annulatus.

    PubMed

    Ravindran, Reghu; Juliet, Sanis; Kumar, K G Ajith; Sunil, A R; Nair, Suresh N; Amithamol, K K; Rawat, Ajay Kumar Singh; Ghosh, Srikanta

    2011-09-01

    The current need of identification of a new acaricidal agent which is acceptable to public as environmentally safe is a daring task. Use of herbal acaricides is such an alternative. Most of the herbal extracts or fractions are dissolved in polar or non-polar solvents or detergents before tested for acaricidal activity. The solvent or detergent to be used for dissolving the herbal extract should be of little acaricidal activity. In the present study, experimentations were carried out on adult engorged female ticks to detect the toxicity of different solvents, viz. hexane, petroleum ether, n-butanol, isopropyl alcohol, chloroform, glycerol, ethyl acetate, acetone, ethanol, and methanol. The study revealed that methanol was the least toxic solvent against Rhipicephalus (Boophilus) annulatus.

  3. Pyrethroid resistance in Iranian field populations of Rhipicephalus (Boophilus) annulatus.

    PubMed

    Ziapour, Seyyed Payman; Kheiri, Sadegh; Fazeli-Dinan, Mahmoud; Sahraei-Rostami, Farzaneh; Mohammadpour, Reza Ali; Aarabi, Mohsen; Nikookar, Seyed Hassan; Sarafrazi, Mohammad; Asgarian, Fatemeh; Enayati, Ahmadali; Hemingway, Janet

    2017-03-01

    Resistance to acaricides in ticks is becoming increasingly widespread throughout the world; therefore, tick control requires resistance monitoring for each tick species. The aims of this study were to monitor the susceptibility status of the cattle tick, Rhipicephalus (Boophilus) annulatus (Acari: Ixodidae), against pyrethroid acaricides from Mazandaran Province, northern Iran, and where resistance was evident, and establish the possible underlying mechanisms. Fully engorged adult R. (B.) annulatus females collected on cattle from Mazandaran Province. Twenty-nine tick populations produced 10-18days old larvae and bioassayed with cypermethrin and λ-cyhalothrin by larval packet test and the levels of detoxification enzymes were measured. Population AM-29 had a maximum resistance ratio (RR99) of 20.21 to cypermethrin and 53.57% of the tick populations were resistant at LC99 level. With λ-cyhalothrin, 17.86% of the tick populations were resistant and AM-29 was the most resistant population with RR99=4.54. AM-29 also showed significant elevation of cytochrome P450 monooxygenase (P450) and glutathione S-transferase (GST) (2.76- and 2.39-fold, respectively) (P<0.001). Rhipicephalus (Boophilus) annulatus showed resistance to pyrethroid insecticides with elevated levels of P450, GST and para-nitrophenyl acetate (p-NPA) in resistant populations. Operational failure was noted in controlling R. (B.) annulatus by pyrethroid insecticides, therefore alternative pest management measures should be adopted in Iran. For the first time, a new estimate of insecticide resistance based on effective dose recommended by the pesticide manufacturer termed Operational Dose Ratio (ODR) is defined and discussed.

  4. In vivo efficacy of a biotherapic and eugenol formulation against Rhipicephalus microplus.

    PubMed

    Valente, Paula Pimentel; Moreira, Gustavo Henrique Ferreira Abreu; Serafini, Matheus Ferreira; Facury-Filho, Elias Jorge; Carvalho, Antônio Último; Faraco, André Augusto Gomes; Castilho, Rachel Oliveira; Ribeiro, Múcio Flávio Barbosa

    2017-03-01

    The control of Rhipicephalus microplus is essential to prevent cattle discomfort and economic losses. However, increased resistance and acaricides inefficiency lead producers to adopt strategies that could result in the accumulation of chemical residues in meat and milk with possibilities of poisoning in animals and people. This scenario demonstrates the necessity of research into the identification of novel, effective and environmentally safe therapeutic options for cattle tick control. The objectives of this study were to develop and assess the efficacy of R. microplus biotherapic and of 5% eugenol for the control of R. microplus in artificially infested calves. Eighteen male 6-month-old Holstein calves were divided into three groups of six animals. In Group 1, the animals did not receive medication (control group); in Group 2, the animals received 1 mL of R. microplus biotherapic at dilution 6CH (centesimal Hahnemannian), orally administered twice daily. And in Group 3, they received a single application of eugenol 5% in the pour-on formulation. The median efficacy for biotherapy and eugenol 5% was respectively 10.13 and 13.97%; however, upon analyzing reproductive efficiency, it is noteworthy that the biotherapic had 45.86% efficiency and was superior to the action of eugenol (12.03%) after 37 days of treatment. The ultrastructural study provided information about the effects of R. microplus biotherapic on the ovaries of engorged females and showed disorganization in the deposition of the oocyte exochorion. The results suggest hatchability inhibition of larvae, interference in R. microplus reproduction and future possibilities for eco-friendly control of R. microplus with biotherapic 6CH.

  5. Minimum infection rate of Ehrlichia minasensis in Rhipicephalus microplus and Amblyomma sculptum ticks in Brazil.

    PubMed

    Carvalho, Izabelle T S; Melo, Andréia L T; Freitas, Leodil C; Verçoza, Rodolfo V; Alves, Alvair S; Costa, Jackeliny S; Chitarra, Cristiane S; Nakazato, Luciano; Dutra, Valéria; Pacheco, Richard C; Aguiar, Daniel M

    2016-07-01

    A new genotype phylogenetically close to Ehrlichia canis named as Ehrlichia minasensis was identified infecting cattle and deer in Canada, as well as Rhipicephalus microplus ticks and cattle in Brazil. Although it was detected in R. microplus, little is known about the epidemiology of this ehrlichiosis, especially in other tick species. This study evaluated the minimum infection rate of E. minasensis in Amblyomma sculptum and R. microplus ticks from locations where naturally infected cattle were previously detected. Overall, 45 engorged R. microplus ticks after molting [43 pools of adults (13.4%), and 2 pools of nymphs (4%)], and 42 engorged females post-oviposition (30.6%) (p=0.008) were positive by PCR for Ehrlichia sp. using the dsb, 16S rRNA and TRP36 genes, making a total of 87 R. microplus samples positive for Ehrlichia spp. (17.1%, IC 95% 14.01-20.75%). The partial sequences generated in the present study were 99-100% similar to the dsb DNA sequence of E. minasensis genotypes UFMG-EV and UFMT-BV, respectively, 100% similar to the 16S rRNA sequence of the E. minasensis genotype BOV2010 from Canada, and 99% similar to the TRP36 sequence of the Ehrlichia sp. UFMT-BV. The results of this study confirm the occurrence of transstadial transmission of this agent in R. microplus ticks and highlight the importance of R. microplus in the epidemiology and transmission of ehrlichiosis in cattle. No A. sculptum ticks were positive by PCR for E. minasensis.

  6. Acetylcholinesterases of Rhipicephalus (Boophilus) microplus – Multiple gene expression presents an opportune model system for elucidation of multiple functions of AChEs.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Acetylcholinesterase (AChE) is a key neural enzyme of both vertebrates and invertebrates, and is the biochemical target of organophosphate and carbamate pesticides for invertebrates, as well as vertebrate nerve agents, e.g., soman, tabun, VX, and others. AChE inhibitors are also key drugs among thos...

  7. The penultimate arginine of the carboxy terminus determines slow desensitisation in a P2X receptor from the cattle tick Boophilus microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    P2X ion channels have been functionally characterised from a range of eukaryotes. Whilst these receptors can be broadly classified into fast and slow desensitising, the molecular mechanisms underlying current desensitisation are not fully understood. Here we describe the characterisation of a P2X ch...

  8. Acaricidal efficacies of Lippia gracilis essential oil and its phytochemicals against organophosphate-resistant and susceptible strains of Rhipicephalus (Boophilus) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plant-derived natural products can serve as an alternative to synthetic compounds for control of ticks of veterinary and medical importance. Lippia gracilis is an aromatic plant that produces essential oil with high content of carvacrol and thymol monoterpenes. These monoterpenes have high acaricida...

  9. Acaricidal activity of eugenol on Rhipicephalus microplus (Acari: Ixodidae) and Dermacentor nitens (Acari: Ixodidae) larvae.

    PubMed

    de Monteiro, Caio Márcio; Maturano, Ralph; Daemon, Erik; Catunda-Junior, Francisco Eduardo Aragão; Calmon, Fernanda; Senra, Tatiane de Souza; Faza, Aline; de Carvalho, Mário Geraldo

    2012-09-01

    The aim of this study was to evaluate the acaricidal activity of eugenol, with different solubilizations and concentrations, on Rhipicephalus microplus and Dermacentor nitens larvae and to determine the lethal time. The study consisted of four experiments, and the mortality was assessed using the larval packet test with adaptations. The mortality observed in the first experiment was 100 % for all the groups treated with eugenol solubilized in different solvents. In the second, the hydroethanolic formulation of eugenol was used, and the mortality rates for R. microplus and D. nitens was 100 % starting from the concentration of 5.0 μl/ml. In the third experiment, the mortality was 100 % for larvae of both R. microplus and D. nitens after 1 h of contact. And in the fourth experiment, the mortality was above 90 % and statistically similar (p > 0.05) for the four methods the test evaluated.

  10. Genetic Mutations Associated with Pesticide Resistance in Rhipicephalus microplus and Haematobia irritans

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A number of gene mutation in various arthropods have been found to be associated with pesticide resistance. Some of these mutations have been found in the two cattle pests, Rhipicephalus microplus and Haematobia irritans. Sodium channel gene mutations have been associated with pyrethroid resistance ...

  11. Molecular and functional characterization of Bm05br antigen from Rhipicephalus microplus.

    PubMed

    Alzugaray, María Fernanda; Parizi, Luís Fernando; Seixas, Adriana; Benavides, Uruguaysito; da Silva Vaz, Itabajara

    2017-02-01

    Rhipicephalus microplus is a cattle-specific tick, causing considerable losses in the livestock industry. The identification of molecules responsible for modulation of host defenses during different parasite stages can help in the development of alternative methods, such as vaccination, to control tick infestations. Hq05, a protein of unknown function identified in the tick Haemaphysalis qinghaiensis, induced a significant protective immune response when used as a vaccine in sheep. In the present study, we investigated Bm05br, the Hq05 homologous gene from R. microplus. Besides H. qinghaiensis, Bm05br homologous found in other tick species such as Rhipicephalus annulatus, Rhipicephalus sanguineus sensu lato, Haemaphysalis longicornis and Ixodes scapularis were comparatively analyzed. Bm05br expression profile in different R. microplus tissues and life-stages was determined by qRT-PCR and Western blot. Bm05br was detected in ovaries, salivary glands and the fat body of both partially and fully engorged females. The highest transcription levels were observed in partially engorged females fat body and salivary glands. Gene knockdown by RNAi reduced egg hatching rate and the weight of tick larvae obtained from treated group, when compared to controls. These results indicate that Bm05br may be involved in R. microplus reproduction. Together with its distribution and high sequence conservation across different tick species, our data suggest Bm05br as a potential antigen for development of a multispecies anti-tick vaccine.

  12. Unveiling the oxidative metabolism of Rhipicephalus microplus (Acari: Ixodidae) experimentally exposed to entomopathogenic fungi.

    PubMed

    Tunholi-Alves, Vinícius Menezes; Tunholi Alves, Victor Menezes; da Silva, Jairo Pinheiro; Nora Castro, Rosane; Salgueiro, Fernanda Barbosa; Perinotto, Wendell Marcelo de Souza; Gôlo, Patrícia Silva; Camargo, Mariana Guedes; Angelo, Isabele da Costa; Bittencourt, Vânia Rita Elias Pinheiro

    2016-10-01

    Rhipicephalus microplus is an important tick in tropical regions due to the high economic losses caused by its parasitism. Metarhizium anisopliae and Beauveria bassiana are well-known entomopathogenic fungi that can afflict R. microplus ticks. The development of new targets and strategies to control this parasite can be driven by studies of this tick's physiology. Recently, it was reported that when exposed to adverse physiological conditions, ticks can activate fermentative pathways, indicating transition from aerobic to anaerobic metabolism. Nevertheless, the precise mechanism by which entomopathogenic fungi influence R. microplus metabolism has not been clarified, limiting understanding of the tick-fungus association. Thus, the present study aimed to evaluate the effect of infection of ticks by M. anisopliae and B. bassiana on the amount of selected carboxylic acids present in the hemolymph, enabling increased understanding of changes previously reported. The results showed preservation in the concentrations of oxalic, lactic, and pyruvic acids in the hemolymph 24 and 48 h after dropping from cattle; while there were variations in the concentration of these carboxylic acids after infection of female ticks to M. anisopliae and B. bassiana. Significant increases were observed in the concentration of oxalic and lactic acids and significant reduction of pyruvic acid for both observation times (24 and 48 h) after infection by entomopathogenic fungi. These results indicate that B. bassiana and M. anisopliae infection alters the basal metabolism of R. microplus females, resulting in the activation of fermentative pathways.

  13. Role of Rhipicephalus microplus cheliceral receptors in gustation and host differentiation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rhipicephalus microplus is considered the most economically important ectoparasite of cattle worldwide. It is known that zebuine breeds of cattle are less susceptible to tick infestation than taurine breeds. Contact chemoreceptors in the cheliceral pit sensilla of ticks respond selectively to phagos...

  14. Gene's Organ Transcriptome of the southern Cattle Fever Tick, Rhipicephalus microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The southern cattle fever tick (SCFT), Rhipicephalus microplus, is considered the most economically important ectoparasite of livestock worldwide and ranks sixth among the most pesticide resistant arthropods globally. This one-host hard tick is a vector of the infectious agents causing bovine babesi...

  15. Assembled contigs of the synganglion transcriptome from an Australian population of the cattle tick, Rhipicephalus microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a collaboration with National Center for Genome Resources and University of Texas at El Paso researchers, we sequenced and assembled the transcriptome of the synganglion of the Texas strain (Deutsch) of the cattle tick Rhipicephalus microplus. This transcriptome contains 43, 468 sequences and wa...

  16. Assembled contigs of the synganglion transcriptome from a Texas population of the cattle tick, Rhipicephalus microplus.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a collaboration with National Center for Genome Resources and University of Texas at El Paso researchers, we sequenced and assembled the transcriptome of the synganglion of the Texas strain (Deutsch) of the cattle tick Rhipicephalus microplus. This transcriptome contains 43, 468 sequences and wa...

  17. Knockdown of the rhipicephalus microplus cytochrome c oxidase subunit III gene is associated with a failure of anaplasma marginale transmission

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rhipicephalus microplus is an obligate hematophagous ectoparasite of cattle and an important biological vector of Anaplasma marginale in tropical and subtropical regions. The primary determinants for Anaplasma transmission are infection of tick gut epithelial cells followed by infection of salivary ...

  18. An intensive search for promising fungal biological control agents of ticks, particularly Rhipicephalus microplus.

    PubMed

    Fernandes, Everton K K; Angelo, Isabele C; Rangel, Drauzio E N; Bahiense, Thiago C; Moraes, Aurea M L; Roberts, Donald W; Bittencourt, Vânia R E P

    2011-12-15

    Entomopathogenic fungi have been investigated worldwide as promising biological control agents of the cattle tick Rhipicephalus microplus. The current study evaluates the virulence of several fungal isolates to R. microplus larva in the laboratory as part of an effort to identify isolates with promise for effective biocontrol of R. microplus in the field. Sixty fungal isolates, encompassing 5 Beauveria spp. and 1 Engyodontium albus (=Beauveria alba), were included in this study. In addition to bioassays, the isolates were characterized morphologically and investigated as to their potential for conidial mass production. These findings were correlated with previous reports on the same fungal isolates of their natural UV-B tolerance (Fernandes et al., 2007), thermotolerance and cold activity (Fernandes et al., 2008), and genotypes (Fernandes et al., 2009). R. microplus larvae obtained from artificially infested calves were less susceptible to Beauveria bassiana infection than ticks acquired from naturally infested cattle from a different location. Isolates CG 464, CG 500 and CG 206 were among the most virulent Beauveria isolates tested in this study. All fungal isolates presented morphological features consistent with their species descriptions. Of the 53 B. bassiana isolates, five (CG 481, CG 484, CG 206, CG 235 and CG 487) had characteristics that qualified them as promising candidates for biological control agents of R. microplus, viz., mean LC(50) between 10(7) and 10(8)conidiaml(-1); produced 5000 conidia or more on 60mm(2) surface area of PDAY medium; and, in comparison to untreated (control) conidia, had the best conidial tolerances to UV-B (7.04 kJ m(-2)) and heat (45°C, 2h) of 50% or higher, and conidial cold (5°C, 15d) activity (mycelial growth) higher than 60%. The current study of 60 Beauveria spp. isolates, therefore, singles out a few (five) with high potential for controlling ticks under field conditions.

  19. Role of Rhipicephalus microplus cheliceral receptors in gustation and host differentiation.

    PubMed

    Ferreira, Lorena Lopes; Soares, Sara Fernandes; de Oliveira Filho, Jaires Gomes; Oliveira, Thaynara Tatielly; Pérez de León, Adalberto A; Borges, Lígia Miranda Ferreira

    2015-04-01

    Rhipicephalus microplus is considered the most economically important ectoparasite of cattle worldwide. It is known that zebuine breeds of cattle are less susceptible to tick infestation than taurine breeds. Contact chemoreceptors in the cheliceral pit sensilla of ticks respond selectively to phagostimulant compounds, however their role in blood feeding relative to host susceptibility to infestation remains to be fully understood. We addressed this topic by conducting taste electrophysiology experiments with cheliceral pit sensilla preparations of R. microplus females. Solutions of five known ixodid tick phagostimulants were tested at different concentrations: sodium (NaCl), and potassium chloride (KCl) (10(-3)-10(-1)M); glucose (10(-4)-10(-1)M); adenosine triphosphate (ATP) (10(-6)-10(-2)M); and reduced l-glutathione (GSH) (10(-6)-10(-2)M). Serum samples from six susceptible animals of the Girolando breed (5/8 Bos indicus×3/8 B. taurus) and six resistant Nelore bovines (pure B. indicus) were also tested. A dose-dependent response of gustatory neurons associated with the chelicerae sensillum to NaCl, glucose, GSH, and ATP were observed. Responses by the cheliceral inner digit pit sensilla of R. microplus to KCl and glucose were also observed and they are reported here for the first time. In addition to an electrophysiological response to known phagostimulants, chemoreceptors in the chelicera of R. microplus responded differently to serum from cattle susceptible and resistant to infestation. The cheliceral pit neurons were more responsive to serum of R. microplus resistant bovines with a higher mean spike frequency (53.5±2spikess(-1)) than to serum samples from susceptible cattle (40.3±2spikess(-1)). The implications of chemosensation during tick blood feeding are discussed.

  20. SNP Analysis Infers that Recombination Is Involved in the Evolution of Amitraz Resistance in Rhipicephalus microplus

    PubMed Central

    Baron, Samantha; van der Merwe, Nicolaas A.; Madder, Maxime; Maritz-Olivier, Christine

    2015-01-01

    Rhipicephalus microplus, better known as the Asiatic cattle tick, is a largely invasive ectoparasite of great economic importance due to the negative effect it has on agricultural livestock on a global scale, particularly cattle. Tick-borne diseases (babesiosis and anaplasmosis) transmitted by R. microplus are alarming as they decrease the quality of livestock health and production. In sub-Saharan Africa, cattle represent a major source of meat and milk, but this region of the world is severely affected by the Rhipicephalus microplus tick. The principal method for tick control is the use of chemical acaricides, notably amitraz, which was implemented in the 1990’s after resistance to other acaricides surfaced. However, the efficiency of chemical control is hindered by an increase in the frequency of mutant resistance alleles to amitraz in tick populations. Presently, the only way to assess amitraz resistance is by means of larval packet tests, but this technique is time-consuming and not particularly cost effective. The main aims of this study were three-fold. First, we attempted to correlate two known SNPs in the octopamine/tyramine (OCT/Tyr) receptor with amitraz resistance in South African field samples of R. microplus. Second, we calculated gametic disequilibrium for these SNPs to determine whether they are randomly associated. Lastly, we conducted a study to assess the evolutionary effects of recombination within the OCT/Tyr receptor. Our results confirmed that the two SNPs are associated with amitraz resistance in the South African tick strain, and that they are in gametic disequilibrium. Additionally, recombination was detected in the OCT/Tyr receptor generating two recombinant haplotypes. These results are of concern to farmers in sub-Saharan Africa, and the emergence of amitraz resistance should be closely monitored in future. Therefore, we present a quick and affordable RFLP based diagnostic technique to assess amitraz resistance in field samples of R

  1. Multiple mutations in the para-sodium channel gene are associated with pyrethroid resistance in Rhipicephalus microplus from the United States and Mexico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rhipicephalus microplus is an invasive tick vector that transmits the protozoan parasites Babesia bovis and B. bigemina, the causative agents of bovine babesiosis (cattle fever). Acaricide resistant R. microplus populations have become a major problem for many cattle producing areas of the world. Py...

  2. An update on distribution models for Rhipicephalus microplus in West Africa.

    PubMed

    De Clercq, Eva M; Estrada-Peña, Agustin; Adehan, S; Madder, Maxime; Vanwambeke, Sophie O

    2013-11-01

    The cattle tick, Rhipicephalus microplus, which reached the West African region approximately 8 years ago, has established viable populations in Côte d'Ivoire and Benin and spread rapidly from the assumed points of introduction. However, existing maps of its distribution range do not agree on the areas at risk, most probably due to suboptimal modelling approaches. Therefore, we undertook a re-investigation of the potential distribution range based on a high-quality dataset from West Africa that includes information on 104 farms located all over Benin. Focussing on climate suitability and applying advanced modelling, a subset of representative and uncorrelated climate variables was selected and fed into Maxent software to obtain an estimate of climate suitability for West Africa. The resulting map was validated using an independent dataset of 13 farms along the apparent distribution edge. The entire southern part of West Africa (covering southern Nigeria, Benin, Togo and Ghana) features high climate suitability for R. microplus. All of Côte d'Ivoire is inside the distribution range of this tick and the southern rim of Burkina Faso is expected to be suitable for the establishment of R. microplus populations. The validation of the distribution, dated one year after the initial field visit, confirmed the predicted distribution range, although a small number of individuals of R. microplus were found north of the predicted limit. These low numbers might indicate that the climate is not suitable for the establishment of a viable tick population. An alternative explanation is the recent introduction by nomadic cattle herds passing through this location. In this region of the world, it is quite common for cattle owners to lead their livestock over distances of more than 500 km in search of food and water.

  3. Synergistic action of fatty acids, sulphides and stilbene against acaricide-resistant Rhipicephalus microplus ticks.

    PubMed

    Arceo-Medina, G N; Rosado-Aguilar, J A; Rodríguez-Vivas, R I; Borges-Argaez, R

    2016-09-15

    Six compounds in a methanolic extract of Petiveria alliacea stem (cis-stilbene; benzyl disulphide; benzyl trisulphide; and methyl esters of hexadecanoic acid, octadecadienoic acid and octadecenoic acid) are known to exercise acaricide activity against cattle tick Rhipicephalus microplus larvae and adults. The synergistic effect of 57 combinations of these six compounds on acaricide activity against R. microplus was evaluated. Larvae immersion tests produced the lethal concentrations needed to kill 50% (LC50) and 99% (LC99) of the population. Adult immersion tests produced rates (%) for mortality, oviposition inhibition and eclosion inhibition. Individually, none of the compounds (1% concentration) exhibited acaricide activity (mortality ≤2.3%). When combined, however, nine mixtures exhibited a synergistic increase in activity, with high mortality rates (≥92%) in larvae. Values for LC50 ranged from 0.07 to 0.51% and those for LC99 from 0.66 to 5.16%. Thirty six compound mixtures had no significant activity (mortality ≤30%) against larvae. Two mixtures exhibited synergism against adults, with high rates (≥92%) of oviposition inhibition. The mixtures based on the benzyl disulphide+benzyl trisulphide pairing produced a synergistic effect against acaricide-resistant R. microplus larva and adults, and are therefore the most promising combination for controlling this ubiquitous ectoparasite.

  4. Reprolysin metalloproteases from Ixodes persulcatus, Rhipicephalus sanguineus and Rhipicephalus microplus ticks.

    PubMed

    Ali, Abid; Tirloni, Lucas; Isezaki, Masayoshi; Seixas, Adriana; Konnai, Satoru; Ohashi, Kazuhiko; da Silva Vaz Junior, Itabajara; Termignoni, Carlos

    2014-08-01

    Metalloproteases (MPs) have been considered essential for blood feeding and other physiological functions in several hematophagous animals, including ticks. We report the characterization of MP sequences of three important ticks from Asia, Africa and America: Ixodes persulcatus (Ip-MPs), Rhipicephalus sanguineus (Rs-MPs) and R. microplus (BrRm-MPs). Amino acid sequence identity between R. microplus and R. sanguineus MPs ranged from 76 to 100 %, and identities among I. persulcatus, I. ricinus and I. scapularis MP sequences ranged from 88 to 97 %. This high sequence identity and typical functional motifs show that all sequences are MPs. The presence of a zinc binding site, a Met-turn and cysteine rich domain at the C-terminal region indicates that these proteins belong to the reproplysin family of MPs. Differences in amino acid sequences of BrRm-MP1, BrRm-MP2, BrRm-MP4 and BrRm-MP5 (from Porto Alegre strain ticks) were 6, 2, 7 and 5 %, respectively, when compared with sequences deposited in GenBank for the same genes from other R. microplus isolates. Analyses of MPs predicted that they have various highly antigenic regions. Semi-quantitative RT-PCR analysis revealed the presence of transcripts in salivary glands of partially and fully fed female ticks. None of these transcripts were observed in males (except BrRm-MP4) and eggs. These enzymes may be functional components required during tick feeding to manipulate host defenses and support tick hematophagy.

  5. The action of two ethyl carbamates on acetylcholinesterase and reproductive organs of Rhipicephalus microplus.

    PubMed

    Prado-Ochoa, M G; Ramírez-Noguera, P; Díaz-Torres, R; Garrido-Fariña, G I; Vázquez-Valadez, V H; Velázquez-Sánchez, A M; Muñoz-Guzmán, M A; Angeles, E; Alba-Hurtado, F

    2014-01-31

    The effects produced by the new synthetic carbamates ethyl-(4-bromophenyl) carbamate and ethyl-(4-chlorophenyl) carbamate on the acetylcholinesterase (AChE) activity, egg structure and reproductive organs of two Rhipicephalus microplus strains were evaluated. Inhibition kinetic parameters showed that the studied carbamates are weak inhibitors and have a low affinity for R. microplus AChE. Histologically, in oocytes from carbamate-treated engorged female ticks, a loss of shape, cytoplasmic vacuoles, decreased chorion deposition, alterations in cytoplasmic granularity and irregular membranes were observed. In oocyte germinal vesicles, a loss of shape, nucleolar fragmentation and membrane alterations with degenerative signs were observed. The ovarian epithelium was vacuolated, flattened, eroded and contained pyknotic nuclei. These alterations were observed from the first day and persisted and increased in severity until day 7 post-treatment. The ovaries from carbamate-treated ticks had fewer stage IV-V oocytes and more stage I-II oocytes. Additionally, eggs produced by the treated ticks had a modified appearance, decreased size, a reduced superficial waxy layer and a loss of viability. The results of this study show that the effects of carbamates on R. microplus were independent of AChE inhibition and show that the morphological alterations in the reproductive organs were due to carbamate actions on the vitellogenesis and viability of the ovarian cells.

  6. In vitro acaricidal efficacy of plant extracts from Brazilian flora and isolated substances against Rhipicephalus microplus (Acari: Ixodidae).

    PubMed

    Valente, Paula Pimentel; Amorim, Juliana Mendes; Castilho, Rachel Oliveira; Leite, Romário Cerqueira; Ribeiro, Múcio Flávio Barbosa

    2014-01-01

    The tick Rhipicephalus microplus causes significant losses in livestock cattle and has developed increasing resistance to the primary acaricides that are used to treat these infections. The objective of this study was to identify new biomolecules or isolated substances showing acaricidal activity from plants. Larval packet tests were conducted to evaluate the effects of 11 species of plants and three isolated substances (betulinic acid, eugenol, and nerolidol) on R. microplus. An adult female immersion test was performed with the substance that showed the highest larvicidal activity, which was evaluated for inhibition of reproduction. Tests using Licania tomentosa, Hymenaea stigonocarpa, Hymenaea courbaril, Stryphnodendron obovatum, Jacaranda cuspidifolia, Jacaranda ulei, Struthanthus polyrhizus, Chrysobalanus icaco, Vernonia phosphorea, Duguetia furfuracea, and Simarouba versicolor extracts as well as the isolated substance betulinic acid indicated lower acaricidal effects on R. microplus larvae. The extract displaying the best larvicidal activity was the ethanolic extract from L. tomentosa at a concentration of 60%, resulting in a mortality rate of 40.3%. However, nerolidol and eugenol showed larvicidal activity, which was highest for eugenol. Nerolidol caused a 96.5% mortality rate in the R. microplus larvae at a high concentration of 30%, and eugenol caused 100% mortality at a concentration of 0.3%. In the adult immersion test, 5% eugenol was identified as a good biomolecule for controlling R. microplus, as demonstrated by its high acaricidal activity and inhibition of oviposition.

  7. Identification of intestinal bacterial flora in Rhipicephalus microplus ticks by conventional methods and PCR-DGGE analysis.

    PubMed

    Xu, Xing-Li; Cheng, Tian-Yin; Yang, Hu; Yan, Fen

    2015-06-01

    In this study, we have analyzed the intestinal microbial flora associated with Rhipicephalus microplus ticks using both culture-dependent and independent methods based on PCR and denaturing gradient gel electrophoresis (PCR-DGGE). The R. microplus ticks were collected from cattle and goats in Jiangxi, Hunan and Guizhou Provinces of China. Three distinct strains of bacteria were isolated using culture-dependent methods: Staphylococcus simulans, Bacillus subtilis and Bacillus flexus strain. Nineteen distinct DGGE bands were found using PCR-DGGE analysis, and their search for identity shows that they belonged to Rickettsiaceae, Xanthomonadaceae, Coxiella sp., Ehrlichia sp., Pseudomonas sp., Ehrlichia sp., Orphnebius sp., Rickettsia peacockii, Bacillus flexus. Rickettsia peacockii and Coxiella genus were the dominant strain of the R. microplus ticks from cattle, Pseudomonas sp. and B. flexus strain were the most common species in all tick samples from goats. Ehrlichia canis were detected only in R. microplus ticks from Yongshun area in Hunan Province. The results indicate that the intestinal microbial diversity of R. microplus ticks was influenced by tick hosts and local differences in the sampling location and these two aspects may affect transmission of pathogen to humans and animals.

  8. Commercial formulation of Metarhizium anisopliae for the control of Rhipicephalus microplus in a pen study.

    PubMed

    Camargo, Mariana G; Marciano, Allan F; Sá, Fillipe A; Perinotto, Wendell M S; Quinelato, Simone; Gôlo, Patrícia S; Angelo, Isabele C; Prata, Márcia C A; Bittencourt, Vânia R E P

    2014-09-15

    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-based formulation was 47.74% and 40.89% in comparison with control and oil control groups, respectively. Changes in the biological parameters of engorged R. microplus females were observed in the first three days after treatment, with a significant reduction in hatching percentage and egg production index. We concluded that Metarril(®) SP Organic plus 10% mineral oil was efficient against R. microplus in pen studies. However, further in vivo studies are required to increase the efficacy and to establish a protocol for the use of this product in the field against the cattle tick.

  9. Toxicity of DMSO, Triton X 100 and Tween 20 against Rhipicephalus (Boophilus) annulatus.

    PubMed

    Ravindran, Reghu; Juliet, Sanis; Gopalan, Ajith Kumar Karapparambu; Kavalimakkil, Amithamol Krishnan; Ramankutty, Sunil Athalathil; Nair, Suresh Narayanan; Narayanan, Priya Manakkulamparambil; Ghosh, Srikanta

    2011-10-01

    The problems associated with synthetic chemical pesticides include resistance, residues, pest resurgence and the adverse effects on non-target organisms. Hence, many ecofriendly newer alternatives are being evaluated among which one of the most important is the use of herbal agents. Many of the plant extracts or its fractions are dissolved in polar or non polar solvents or detergents before tested for acaricidal activity. The solvent or detergent used for dissolving the herbal extract should have little or no acaricidal effects. In the present study, laboratory tests were carried out on engorged female Rhipicephalus (Boophilus) annulatus ticks to detect the toxicity of common diluents such as dimethyl sulfoxide (DMSO), Tween 20 and Triton X 100 at 1% concentration. The result of the study revealed that least acaricidal activity was with Triton X 100 while the other two inhibited the hatching of eggs laid by treated ticks.

  10. Adaptation of in vitro bioassays for the diagnosis of resistance to Fipronil and Ivermectin in R. (B.) microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    R. microplus represents the most important pathological constraint to livestock production in Brazil and Uruguay. The infestation of cattle by ticks is controlled by chemical applications on a regular basis. Fipronil and ivermectin have been widely used in recent years to the benefit of cattle produ...

  11. Acetylcholinesterase 1 in populations of organophosphate-resistant North American strains of the cattle tick, Rhipicephalus microplus (Acari: Ixodidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rhipicephalus microplus, the cattle fever tick, is a global economic problem to the cattle industry due to direct infestation of cattle and pathogens transmitted during feeding. Cattle fever tick outbreaks continue to occur along the Mexico-U.S. border even though the tick has been eradicated from t...

  12. Acetylcholinesterase 1 in populations of organophosphate resistant North American strains of the cattle tick, Rhipicephalus microplus (Acari: Ixodidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a collaboration with Purdue University researchers, we sequenced a 143,606 base pair Rhipicephalus microplus BAC library clone that contained the coding region for acetylcholinesterase 1 (AChE1). Sequencing was by Sanger protocols and the final assembly resulted in 15 contigs of varying length, e...

  13. Assembled contigs of the Haller's organ transcriptome from an Australian population of the cattle tick, Rhipicephalus microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In a collaboration with National Center for Genome Resources and University of Texas at El Paso researchers, we sequenced and assembled the transcriptome of the Haller's organ of an Australian strain (NRFS) of the cattle tick Rhipicephalus microplus (recently reclassified as Rhipicephalus australis...

  14. Acaricide resistance and strategies to mitigate economic impact of the southern cattle fever tick (Rhipicephalus microplus) on livestock production systems in the Americas

    Technology Transfer Automated Retrieval System (TEKTRAN)

    BACKGROUND: The southern cattle fever tick (SCFT), Rhipicephalus microplus, is considered the most economically important external parasite of livestock worldwide. SCFT populations resistant to acaricides complicate efforts to enhance the productivity of livestock. Here, acaricide resistance is summ...

  15. In vitro cultivation and cryopreservation of Babesia bigemina sporokinetes in hemocytes of Rhipicephalus microplus.

    PubMed

    de Rezende, Jania; Rangel, Charles P; McIntosh, Douglas; Silveira, Júlia A G; Cunha, Nathalie C; Ramos, Carlos A N; Fonseca, Adivaldo H

    2015-09-15

    Cultures of tick hemocytes represent alternative cell lines for the isolation and cultivation of a variety of hemoparasites. The present study reports the development and evaluation of methods for the in vitro culture and maintenance of sporokinetes of Babesia bigemina in association with hemocytes of the tick Rhipicephalus microplus. Hemolymph, from engorged females infected with B. bigemina sporokinetes, was incubated at 28 °C in L15 culture medium supplemented with 40% fetal bovine serum. Adherence of hemocytes to flask surfaces and the development of B. bigemina sporokinetes commenced on the first day of cultivation. The protozoa demonstrated clear motility and the capacity to adhere to hemocyte membranes for up to 25 days, at which time the hemocytes began to show signs of degeneration. Examination of Giemsa stained hemocyte cultures, revealed the presence of pyriformis forms, as well as mature and immature sporokinetes with dark red nuclei, centralized or near the apical extremities. Sporokinetes harvested from culture supernatants were cryopreserved in liquid nitrogen. Inoculation of parasite-free hemocyte cultures with defrosted sporokinetes, demonstrated the viability and interaction of the protozoa with the hemocytes over 21 days. Cultured hemocytes of R. microplus hold potential for development as a tool in the study of host parasite interactions and as a substrate for the in vitro maintenance of B. bigemina sporokinetes.

  16. Influence of tea tree oil (Melaleuca alternifolia) on the cattle tick Rhipicephalus microplus.

    PubMed

    Pazinato, Rafael; Klauck, Vanderlei; Volpato, Andreia; Tonin, Alexandre A; Santos, Roberto C; de Souza, Márcia E; Vaucher, Rodrigo A; Raffin, Renata; Gomes, Patrícia; Felippi, Candice C; Stefani, Lenita M; Da Silva, Aleksandro S

    2014-05-01

    The aim of this study was to verify the influence of tea tree oil (TTO) (Melaleuca alternifolia) tested in its pure and nanostructured (TTO nanoparticles) forms on the reproduction of female Rhipicephalus microplus. For our purpose, female ticks were collected from naturally infected animals and treated in vitro with TTO (1, 5, and 10 %) and TTO nanoparticles (0.075, 0.375, and 0.75 %). In order to validate the tests, they were performed in triplicate using positive (amitraz) and negative (untreated) controls. It was possible to observe that pure TTO (5 and 10 %) and TTO nanoparticles (0.375 and 0.75 %) showed 100 % reproductive inhibition on female ticks. Additionally, pure TTO (1 %) also showed an acaricide effect (70 %), similarly to the positive control (78.3 %). This is the first study demonstrating the activity of pure TTO and TTO nanoparticles on female ticks. Therefore, based on these results, we were able to show that both forms and all concentrations of M. alternifolia affected tick reproduction by inhibiting egg laying and hatching. We were also able to show that TTO nanoparticles potentiated the inhibitor effect of pure TTO on the reproduction of R. microplus.

  17. First report of pyrethroid resistance in Rhipicephalus (Boophilus) annulatus larvae (Say, 1821) from Iran.

    PubMed

    Ziapour, Seyyed Payman; Kheiri, Sadegh; Asgarian, Fatemeh; Fazeli-Dinan, Mahmoud; Yazdi, Fariborz; Mohammadpour, Reza Ali; Aarabi, Mohsen; Enayati, Ahmadali

    2016-04-01

    Rhipicephalus (Boophilus) annulatus is one of the most important hard ticks parasitizing cattle in northern Iran. The aim of this study was to evaluate pyrethroid resistance levels of this species from Nur County, northern Iran. The hard ticks were collected through a multistage cluster randomized sampling method from the study area and fully engorged female R. (B.) annulatus were reared in a controlled insectary until they produced larvae for bioassay. Seventeen populations of the hard ticks were bioassayed with cypermethrin and 12 populations with lambda-cyhalothrin using a modified larval packet test (LPT). Biochemical assays to measure the contents/activity of different enzyme groups including mixed function oxidases (MFOs), glutathione S-transferases (GSTs) and general esterases were performed. Population 75 showed a resistance ratio of 4.05 with cypermethrin when compared with the most susceptible field population 66 at the LC50 level. With lambda-cyhalothrin the resistance ratio based on LC50 was 3.67 when compared with the susceptible population. The results of biochemical assays demonstrated significantly elevated levels of GSTs and esterases in populations tested compared with the heterozygous susceptible filed population and a correlation coefficient of these enzymes was found in association to lambda-cyhalothrin resistance. Based on the results, pyrethroid acaricides may operationally fail to control R. (B.) annulatus in North of Iran. This study is the first document of pyrethroid resistance in R. (B.) annulatus populations from Iran.

  18. Acaricidal activity of alkaloid fractions of Leucas indica Spreng against Rhipicephalus (Boophilus) annulatus tick.

    PubMed

    Divya, T M; Soorya, V C; Amithamol, K K; Juliet, S; Ravindran, R; Nair, S N; Ajithkumar, K G

    2014-03-01

    The acaricidal activity from alkaloid and non-alkaloid fractions of Leucas indica were studied against Rhipicephalus (Boophilus) annulatus tick using adult immersion test under laboratory conditions. For this purpose, the engorged female R.(B.) annulatus tick were exposed to two fold serial dilutions of alkaloid extract (50 mg/ml, 25 mg/ml, 12.5 mg/ml, 6 mg/ml and 3 mg/ml) using 'dipping method' in vitro. The efficacy was assessed by measuring the percentage of adult mortality, inhibition of fecundity and hatching rate. The alkaloid fraction of the extract produced concentration dependent delayed adult tick mortality. The extract at a concentration of 50 mg/ml demonstrated 66.67 per cent mortality and 55.16 per cent inhibition of fecundity. Nicotine was identified as one of the compounds of alkaloid fraction. However, it did not reveal any acaricidal activity when tested in vitro at concentrations ranging from 62.5-1000 μg/mL. Hence, the acaricidal action of L. indica is not due to nicotine. Non alkaloid fraction also did not reveal any acaricidal effects against R. (B.) annulatus tick.

  19. Identification of Tropomyosin and Its Immunological Properties from Larvae of Cattle Tick, Boophilus annulatus

    PubMed Central

    Nabian, S; Taheri, M; Fard, R Mazaheri Nezhad; Aramoon, M

    2013-01-01

    Background Boophilus annulatus is an obligate blood feeder tick that can cause great losses in animals due to anemia and its ability to injure its host skin directly. The aim of this study was identification of cattle humoral immune response to some tick proteins during experimental infestation. Methods Immune sera against tick were collected from experimentally infested cattle with ticks. One and two-dimensional electrophoresis and Western blotting methods were used for the detection of immunogenic proteins in larval tick extract and eight of these proteins were identified by MALDITOF and MALDI-TOF-TOF mass spectrometry. Results In non-reducing one-dimensional SDS-PAGE, some bounds between 12 to more than 250-kDa appeared. In two-dimensional SDS-PAGE, numerous spot appeared and the identified immunogenic proteins by parallel immunoblotting weighted between 14 and 97 kDa. Amino acid sequences of protein spot with 37-kDa molecular weight had identity to tropomyosin based on Mascot search in NCBI. Conclusion Anti tropomyosin antibodies can be induced in experimentally infested hosts with ticks and it seems that tropomyosin can be useful for the development of anti tick vaccines. PMID:23914237

  20. Acaricidal activity of thymol against larvae of Rhipicephalus microplus (Acari: Ixodidae) under semi-natural conditions.

    PubMed

    Araújo, Laryssa Xavier; Novato, Tatiane Pinheiro Lopes; Zeringota, Viviane; Matos, Renata Silva; Senra, Tatiane Oliveira Souza; Maturano, Ralph; Prata, Márcia Cristina Azevedo; Daemon, Erik; Monteiro, Caio Márcio Oliveira

    2015-09-01

    This is the first study to investigate the activity of thymol on Rhipicephalus microplus larvae under semi-natural conditions. For this purpose, tests were conducted in pots with Brachiaria decumbens seedlings containing cattle tick larvae. Thymol, diluted in ethanol 50° GL, was tested at concentrations of 2.5, 5.0, 10.0, 15.0, and 20.0 mg/mL, along with the control group treated with the solvent alone. Each treatment was composed of five pots (1 pot = a repetition). The experiment was performed in three steps. On the first day, the larvae were applied at the base of the signalgrass. Twenty-four hours later, approximately 25 mL of the solution was applied with thymol on the top of the vegetation in each pot. The survival of the larvae was measured 24 h after application of the solutions. Each pot was analyzed individually, and the grass fillets contained larvae were cut with scissors, placed in Petri dishes, and taken to the laboratory to count the number of living larvae. At the highest concentrations (10, 15, and 20 mg /mL), the number of live larvae declined by more than 95 % in relation to the control group. The lethal concentration 50 % (LC50) and LC90 values were 3.45 and 9.25 mg/ml, respectively. The application of thymol in semi-natural conditions starting concentration of 10 mg/mL significantly reduced the number of living R. microplus larvae.

  1. Evaluation of tea tree oil for controlling Rhipicephalus microplus in dairy cows.

    PubMed

    Pazinatto Boito, Jhonatan; Santos, Roberto C; Vaucher, Rodrigo A; Raffin, Renata; Machado, Gustavo; Tonin, Alexandre A; Da Silva, Aleksandro S

    2016-07-30

    Our research aimed to test the effects of Melaleuca alternifolia oil (pure and in nanocapsules) in the control of Rhipicephalus microplus in dairy cattle. For this purpose, the in vivo studies used 15 cows distributed in three different groups with the same number of animals. Five cows remained untreated (Group A), representing the control group; other five cows were sprayed with TTO (at 5%) in its pure form (Group B); and five cows were sprayed with nanocapsules of TTO (at 0.75%) (Group C). On days 1 and 4 post-treatments (PT), all cows had their ticks counted. On day 1 PT, two ticks from each cow were collected to evaluate the effect of the treatment on ticḱs reproduction (in vitro assays). The pure form of TTO caused a significant reduction (P<0.05) in the number of ticks from the Group B compared to the Group A on day 4 PT. However, there was no significant difference in the number of ticks on cows from Groups A and C after treatment (P>0.05). Treatment with TTO in nanocapsules (Group C) interfered with R. microplus reproduction, leading to lower oviposition by female ticks and hatchability (34.5% of efficacy). On the other hand, TTO oil (Group B) did not interfere on ticḱs reproduction, i.e. showed higher hatchability than the control group. Therefore, it is possible to conclude that pure TTO has an acaricidal effect in dairy cows, in addition to an effect on ticḱs reproduction when used its nanocapsulated form.

  2. Synergism of thymol, carvacrol and eugenol in larvae of the cattle tick, Rhipicephalus microplus, and brown dog tick, Rhipicephalus sanguineus.

    PubMed

    Araújo, L X; Novato, T P L; Zeringota, V; Maturano, R; Melo, D; DA Silva, B C; Daemon, E; DE Carvalho, M G; Monteiro, C M O

    2016-12-01

    The effects of combinations of the monoterpenes thymol and carvacrol and the phenylpropanoid eugenol in larvae of Rhipicephalus microplus (Canestrini, 1888) (Acari: Ixodidae) and Rhipicephalus sanguineus sensu lato (s.l.) (Acari: Ixodidae) were assessed by the larval packet test. The CompuSyn program was used to make qualitative assessments of the effects (synergistic, additive and antagonistic) of the associations. The effects of all combinations tested against R. microplus larvae were synergistic, with combination indices (CIs) <0.70. When tested against R. sanguineus, eight of the mixtures showed a synergistic effect (CI < 0.70); only the carvacrol + thymol mixture at LC50 presented a moderate synergistic effect, with CIs between 0.70-0.90. This study is the first to determine the effects of the interactions of these substances in the control of these two tick species. The combinations of carvacrol + thymol, carvacrol + eugenol and thymol + eugenol have synergistic effects in R. microplus and R. sanguineus s.l. larvae.

  3. Assessment of the repellent effect of Lippia alba essential oil and major monoterpenes on the cattle tick Rhipicephalus microplus.

    PubMed

    Lima, A da Silva; Carvalho, J F de; Peixoto, M G; Blank, A F; Borges, L M F; Costa Junior, L M

    2016-03-01

    The control of Rhipicephalus microplus (Ixodida: Ixodidae) is achieved using synthetic acaricides. However, resistant tick populations are widespread around the world. Plant essential oils can act as repellents, keeping ticks away from hosts and decreasing the selection pressure on synthetic acaricides. The aim of this study was to evaluate the in vitro repellent effect of Lippia alba essential oil on R. microplus larvae. Leaves from two L. alba genotypes maintained under the same agronomic and environmental conditions were collected. Essential oil was extracted by hydrodistillation and analysed by gas chromatography-mass spectrometry (GC-MS). The major monoterpenes detected in the chemical analysis were commercially acquired and tested. For the repellency test, a glass rod was vertically fixed to measure active climbing of approximately 30 R. microplus larvae aged 14-21 days in response to essential oils and monoterpenes. Repellency was evaluated at 1 h, 3 h and 5 h after treatment. Variation in repellent action was detected between the genotypes. The major monoterpenes identified in the essential oils (limonene and carvone) showed low repellent effects in comparison with intact essential oils. Thus, the present results showed that L. alba essential oil contains bioactive compounds with great repellent activity against ticks that varies according to the plant genotype.

  4. Artificial feeding of Rhipicephalus microplus female ticks with anti calreticulin serum do not influence tick and Babesia bigemina acquisition.

    PubMed

    Antunes, Sandra; Merino, Octávio; Lérias, Joana; Domingues, Nuno; Mosqueda, Juan; de la Fuente, José; Domingos, Ana

    2015-02-01

    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 B. bigemina infection in R. annulatus ticks, was expressed as recombinant protein (rCRT) in an E. coli expression system and antibodies raised against rCRT. Anti-rCRT serum was supplemented to a blood meal, offered to partially engorged R. microplus females and their effect in feeding process as well as infection by B. bigemina was analyzed. No significant reductions in tick and egg weight were observed when ticks fed with anti-rCRT serum. Furthermore, B. bigemina infection levels did not show a statistically significant decrease when ticks fed with anti-rCRT antibodies. Results suggest that CRT is not a suitable candidate for cattle vaccination trials.

  5. Comparison of insect kinin analogs with cis-peptide bond, type VI-turn motifs identifies optimal stereochemistry for interaction with a recombinant arthropod kinin receptor from the Southern cattle tick, Boophilus microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The multifunctional ‘insect kinins’ share the evolutionarily conserved C-terminal pentapeptide motif Phe-X1-X2-Trp-Gly-NH2, where X1 = His, Asn, Ser, or Tyr and X2 = Ser, Pro, or Ala; and are associated with the regulation of diuresis in a variety of species of insects. We previously reported the f...

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

    PubMed

    Fernández-Salas, A; Alonso-Díaz, M A; Alonso-Morales, R A; Lezama-Gutiérrez, R; Rodríguez-Rodríguez, J C; Cervantes-Chávez, J A

    2017-03-01

    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 × 10(8) 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.

  7. Molecular cloning, expression and characterization of a functional GSTmu class from the cattle tick Boophilus annulatus.

    PubMed

    Shahein, Yasser Ezzat; El Sayed El-Hakim, Amr; Abouelella, Amira Mohamed Kamal; Hamed, Ragaa Reda; Allam, Shaimaa Abdul-Moez; Farid, Nevin Mahmoud

    2008-03-25

    A full-length cDNA of a glutathione S-transferase (GST) was cloned from a cDNA library of the local Egyptian cattle tick Boophilus annulatus. The 672 bp cloned fragment was sequenced and showed an open reading frame encoding a protein of 223 amino acids. Comparison of the deduced amino acid sequence with GSTs from other species revealed that the sequence is closely related to the mammalian mu-class GST. The cloned gene was expressed in E. coli under T7 promotor of pET-30b vector, and purified under native conditions. The purified enzyme appeared as a single band on 12% SDS-PAGE and has a molecular weight of 30.8 kDa including the histidine tag of the vector. The purified enzyme was assayed upon the chromogenic substrate 1-chloro-2,4-dinitrobenzene (CDNB) and the recombinant enzyme showed high level of activity even in the presence of the beta-galactosidase region on its 5' end and showed maximum activity at pH 7.5. The Km values for CDNB and GSH were 0.57 and 0.79 mM, respectively. The over expressed rBaGST showed high activity toward CDNB (121 units/mg protein) and less toward DCNB (29.3 units/mg protein). rBaGST exhibited peroxidatic activity on cumene hydroperoxide sharing this property with GSTs belonging to the GST alpha class. I50 values for cibacron blue and bromosulfophthalein were 0.22 and 8.45 microM, respectively, sharing this property with the mammalian GSTmu class. Immunoblotting revealed the presence of the GST molecule in B. annulatus protein extracts; whole tick, larvae, gut, salivary gland and ovary. Homologues to the GSTmu were also detected in other tick species as Hyalomma dromedarii and Rhipicephalus sp. while in Ornithodoros moubata, GSTmu homologue could not be detected.

  8. Molecular cloning and expression of a larval immunogenic protein from the cattle tick Boophilus annulatus.

    PubMed

    Shahein, Yasser Ezzat

    2008-02-15

    A full-length cDNA of an immunogenic protein was cloned from a cDNA library of the local Egyptian cattle tick Boophilus annulatus. Antibodies raised against B. annulatus larval proteins were used to screen a cDNA expression library. A 936bp cloned fragment was sequenced and showed an open reading frame of 516bp encoding a protein of 171 amino acids. Comparison of the deduced amino acid sequence with protein data bank revealed that the sequence is related to a sequence isolated from the hard tick Haemaphysalis qinghaiensis (Hq05). Southern blot analysis of B. annulatus genomic DNA showed that the cloned cDNA hybridized to double bands per restriction digest, suggesting that the cloned cDNA is a double copy gene. Amino acid analysis of the cloned gene revealed the presence of two casein kinase II phosphorylation sites in the N-terminal domain suggesting that this molecule may be involved in the signal transduction or gene expression pathways. RT-PCR and northern blotting revealed the presence of two isoforms of the Ba05 gene in salivary glands and in the 3-day-old eggs. The cloned gene without the signal peptide, was expressed in Escherichia coli under T7 promotor of pET-30b vector, and purified under denaturation conditions. The purified protein appeared as a single band on 12% SDS-PAGE with a molecular weight around 22.8kDa including the histidine tag of the vector. Antibodies raised against the purified molecule were used to detect the B. annulatus homologue to the Hq05 gene in whole tick, larvae and gut protein extracts. Immunoblotting revealed the presence of this molecule Ba05 only in whole tick and larval protein extracts and not in the gut protein extract. Using the same antibodies, homologues to the Ba05 gene were detected in other tick species as Hyalomma dromedarii and Rhipicephalus sp. but not in Ornithodoros moubata.

  9. The host status of African buffaloes, Syncerus caffer, for Rhipicephalus (Boophilus) decoloratus.

    PubMed

    Horak, I G; Golezardy, H; Uys, A C

    2006-09-01

    The objective of this study was to assess the host status of African buffaloes, Syncerus caffer, for the one-host tick Rhipicephalus (Boophilus) decoloratus. To this end the R. (B.) decoloratus burdens of ten buffaloes examined in three north-eastern KwaZulu-Natal Province (KZN) nature reserves were compared with those of medium-sized to large antelope species in these reserves and in the southern Kruger National Park (KNP), Mpumalanga Province. The R. (B.) decoloratus burdens of the buffaloes were considerably smaller than those of the antelopes in the KNP, but not those in the KZN reserves. The life-stage structure of the R. (B.) decoloratus populations on the buffaloes, in which larvae predominated, was closer to that of this tick on blue wildebeest, Connochaetes taurinus, a tick-resistant animal, than to that on other antelopes. A single buffalo examined in the KNP was not infested with R. (B.) decoloratus, whereas a giraffe, Giraffa camelopardalis, examined at the same locality and time, harboured a small number of ticks. In a nature reserve in Mpumalanga Province adjacent to the KNP, two immobilized buffaloes, from which only adult ticks were collected, were not infested with R. (B.) decoloratus, whereas greater kudus, Tragelaphus strepsiceros, examined during the same time of year in the KNP harboured large numbers of adult ticks of this species. African buffaloes would thus appear to be resistant to infestation with R. (B.) decoloratus, and this resistance is expressed as the prevention of the majority of tick larvae from developing to nymphs.

  10. Effect of Allium sativum and Allium cepa oils on different stages of Boophilus annulatus.

    PubMed

    Aboelhadid, S M; Kamel, A A; Arafa, W M; Shokier, K A

    2013-05-01

    The study is aimed to investigate the acaricidal effect of Allium sativum (garlic) and Allium cepa (onion) oils on different stages of Boophilus annulatus hard tick. Engorged B. annulatus females were collected from naturally infected cattle. A number of engorged ticks were incubated at 28 °C and 85 % relative humidity to lay eggs, which were incubated to obtain larvae that were used in the study. The used garlic and onion oils were prepared by steam distillation and were analyzed by gas chromatography. These oils were dissolved in ethanol, methanol alcohols, and, partially, in water. The oils were tested in different concentrations; 1, 2, 5, 10, and 20 %. These concentrations were applied on adult ticks by adult immersion test; on larvae by larval immersion technique and on eggs. The 20, 10, and 5 % of garlic oil dilutions in ethanol and methanol killed all adult ticks and larvae within 24 h. Similar results were obtained for 10 and 20 % garlic oil dissolved in water. The effect of 10 % aqueous solution of garlic oil on embryonated eggs was clear as its addition to these eggs led to their in ability to hatch, deformity in shape, and change in color. The 10 and 20 % onion oil in ethanol and methanol alcohols killed 76-86 % of the adult ticks within 72 h post-application. While, all larvae died within 24 h postsubjected to these two concentrations. These concentrations (10 and 20 %) of onion oil in water killed 56-80 % of the treated ticks. Moreover, 10 % aqueous solution of onion oil prevented hatching of embyonated eggs. We concluded that garlic and onion oils have acaricidal effect on all stages of B. annulatus at concentrations higher than 5 %. Only garlic oil could kill 100 % of adult ticks at concentrations from 5 % in alcohols.

  11. Use of Ivermectin as Endoparasiticide in Tropical Cattle Herds Generates Resistance in Gastrointestinal Nematodes and the Tick Rhipicephalus microplus (Acari: Ixodidae).

    PubMed

    Alegría-López, M A; Rodríguez-Vivas, R I; Torres-Acosta, J F J; Ojeda-Chi, M M; Rosado-Aguilar, J A

    2015-03-01

    The objective of the present study was to determine simultaneously the status of resistance against ivermectin (IVM) in gastrointestinal nematodes (GIN) and Rhipicephalus microplus (Canestrini, 1888) ticks in 12 cattle farms where IVM was used for the control of GIN in the Mexican tropics. Six farms had frequent use of IVM (≥ 4 times per year) and six farms had low frequency of IVM use (1-2 times per year). The fecal egg count reduction test and the larval immersion test were used to determine the resistant status of GIN and R. microplus against IVM, respectively. The results indicated that 100% of the surveyed farms had IVM-resistant GIN (reduction % from 0 to 67%). The genera involved were Haemonchus, Cooperia, Ostertagia, Trichostrongylus, and Oesophagostomum. Although the IVM was never used for the control of ticks, 50% of the surveyed farms presented GIN and R. microplus simultaneously resistant to IVM. Furthermore, two R. microplus populations showed high resistance ratio (RR) to IVM (farm TAT: RR50% = 7 and RR99% = 40.1; and farm SLS: RR50% = 2.4; RR99% = 11.0). A high frequency of IVM use (≥ 4 times per year) seemed to promote IVM resistance amongst R. microplus ticks compared with the farms with low frequency of IVM use (1-2 times per year; 66.6 vs. 25.0%, respectively). However, the number of surveyed farms was insufficient to show clear statistical inferences (odds ratio = 6.00; 95% CI = 0.341-105.5). The use of IVM for the control of GIN promoted simultaneously the development of IVM resistance in the GIN and R. microplus populations of the cattle herds surveyed.

  12. Genetic diversity, acaricide resistance status and evolutionary potential of a Rhipicephalus microplus population from a disease-controlled cattle farming area in South Africa.

    PubMed

    Robbertse, Luïse; Baron, Samantha; van der Merwe, Nicolaas A; Madder, Maxime; Stoltsz, Wilhelm H; Maritz-Olivier, Christine

    2016-06-01

    The Southern cattle tick, Rhipicephalus microplus is a hematophagous ectoparasite of great veterinary and economic importance. Along with its adaptability, reproductive success and vectoring capacity, R. microplus has been reported to develop resistance to the major chemical classes of acaricides currently in use. In South Africa, the Mnisi community in the Mpumalanga region offers a unique opportunity to study the adaptive potential of R. microplus. The aims of this study therefore included characterising acaricide resistance and determining the level and pattern of genetic diversity for R. microplus in this region from one primary population consisting of 12 communal dip-stations. The level of acaricide resistance was evaluated using single nucleotide polymorphisms (SNPs) in genes that contribute to acaricide insensitivity. Additionally, the ribosomal internal transcribed spacer 2 (ITS2) gene fragments of collected individuals were sequenced and a haplotype network was constructed. A high prevalence of alleles attributed to resistance against formamidines (amitraz) in the octopamine/tyramine (OCT/Tyr) receptor (frequency of 0.55) and pyrethroids in the carboxylesterase (frequency of 0.81) genes were observed. Overall, the sampled tick population was homozygous resistant to pyrethroid-based acaricides in the voltage-gated sodium channel (VGS) gene. A total of 11 haplotypes were identified in the Mnisi R. microplus population from ITS2 analysis with no clear population structure. From these allele frequencies it appears that formamidine resistance in the Mnisi community is on the rise, as the R. microplus populations is acquiring or generating these resistance alleles. Apart from rearing multi-resistant ticks to commonly used acaricides in this community these ticks may pose future problems to its surrounding areas.

  13. Acaricide and ivermectin resistance in a field population of Rhipicephalus microplus (Acari: Ixodidae) collected from red deer (Cervus elaphus) in the Mexican tropics.

    PubMed

    Rodríguez-Vivas, R I; Miller, R J; Ojeda-Chi, M M; Rosado-Aguilar, J A; Trinidad-Martínez, I C; Pérez de León, A A

    2014-02-24

    In the Neotropics the control of tick infestations in red deer (Cervus elaphus) is achieved primarily through the use of acaricides and macrocyclic lactones. In Mexico, resistance to one or multiple classes of acaricides has been reported in Rhipicephalus microplus infesting cattle, but information on acaricide susceptibility in R. microplus infesting red deer is lacking. In this study we report the level of resistance to different classes of acaricides and ivermectin in R. microplus collected from red deer in the Mexican tropics. Engorged R. microplus females were collected from a red deer farm in Yucatan, Mexico. The larval packet test was used to detect resistance to the organophosphates (OPs) chlorpyrifos and coumaphos, synthetic pyrethroids (SPs) cypermethrin and permethrin, and the phenylpyrazol, fipronil. Resistance to the formamidine amitraz (Am), and ivermectin was ascertained using the larval immersion test. Data were subjected to probit analysis to determine lethal concentrations and resistance ratios to kill 50% (RR50) and 99% (RR99) of the tick population under evaluation in relation to susceptible reference strains. Additionally, allele specific polymerase chain reaction was used to detect the sodium channel F1550I mutation associated with SP resistance in R. microplus. The R. microplus population from red deer in Yucatan showed very high resistance to the two SPs evaluated (RRs>72.2 for cypermethrin; RR for permethrin resistance was so high a dose-response curve was not possible). All individual larvae tested to detect the sodium channel F1550I mutation associated with SP resistance in R. microplus were homozygous. The same tick population showed different levels of resistance to OPs (chlorpyrifos: RR50=1.55, RR99=0.63; coumaphos: RR50=6.8, RR99=5.9), fipronil (RR50=1.8, RR99=0.9), and amitraz (RR50=2.3, RR99=4.4). Resistance to ivermectin was regarded as moderate (RR50=7.1, RR99=5.0). This is the first report of R. microplus ticks collected from red

  14. Adrenergic ligands that block oviposition in the cattle tick Rhipicephalus microplus affect ovary contraction.

    PubMed

    Cossío-Bayúgar, Raquel; Miranda-Miranda, Estefan; Fernández-Rubalcaba, Manuel; Narváez Padilla, Verónica; Reynaud, Enrique

    2015-10-12

    The tyraminergic/octopaminergic system is central for the control of arthropod oviposition. Previous works demonstrated that the pharmacological perturbation of this system inhibits oviposition in the cattle tick Rhipicephalus microplus. In this work, we describe a physiologically active whole-mount preparation of the contractile tick ovary that allows the quantitative videometrical analysis of ovary contraction in response to different compounds. Eight adrenergic ligands known to inhibit oviposition, including octopamine and tyramine were tested. These compounds exhibited antagonistic effects; octopamine relaxes the ovary preparation while tyramine induces a very strong contraction. The other adrenergic compounds tested were classified as able to contract or relax ovary muscle tissue. Isoprotenerol has a stronger relaxative effect than octopamine. Tyramine induces the biggest contraction observed of all the compounds tested, followed, in descending amount of contraction, by salbutamol, prazosin, epinastine, clonidine and the acaricide amitraz. The effect of these adrenergic ligands on the ovary preparation, explains why these molecules inhibit tick oviposition and suggest a regulatory mechanism for ovary contraction and relaxation during oviposition. Our results also provide a physiological explanation of the egg-laying inhibition effect of amitraz when used on the cattle tick.

  15. Adrenergic ligands that block oviposition in the cattle tick Rhipicephalus microplus affect ovary contraction

    PubMed Central

    Cossío-Bayúgar, Raquel; Miranda-Miranda, Estefan; Fernández-Rubalcaba, Manuel; Narváez Padilla, Verónica; Reynaud, Enrique

    2015-01-01

    The tyraminergic/octopaminergic system is central for the control of arthropod oviposition. Previous works demonstrated that the pharmacological perturbation of this system inhibits oviposition in the cattle tick Rhipicephalus microplus. In this work, we describe a physiologically active whole-mount preparation of the contractile tick ovary that allows the quantitative videometrical analysis of ovary contraction in response to different compounds. Eight adrenergic ligands known to inhibit oviposition, including octopamine and tyramine were tested. These compounds exhibited antagonistic effects; octopamine relaxes the ovary preparation while tyramine induces a very strong contraction. The other adrenergic compounds tested were classified as able to contract or relax ovary muscle tissue. Isoprotenerol has a stronger relaxative effect than octopamine. Tyramine induces the biggest contraction observed of all the compounds tested, followed, in descending amount of contraction, by salbutamol, prazosin, epinastine, clonidine and the acaricide amitraz. The effect of these adrenergic ligands on the ovary preparation, explains why these molecules inhibit tick oviposition and suggest a regulatory mechanism for ovary contraction and relaxation during oviposition. Our results also provide a physiological explanation of the egg-laying inhibition effect of amitraz when used on the cattle tick. PMID:26456007

  16. Inhibition of the recombinant cattle tick Rhipicephalus (Boophilus) annulatus glutathione S-transferase.

    PubMed

    Guneidy, Rasha A; Shahein, Yasser E; Abouelella, Amira M K; Zaki, Eman R; Hamed, Ragaa R

    2014-09-01

    Rhipicephalus (Boophilus) annulatus is a bloodsucking ectoparasite that causes severe production losses in the cattle industry. This study aims to evaluate the in vitro effects of tannic acid, hematin (GST inhibitors) and different plant extracts (rich in tannic acid) on the activity of the recombinant glutathione S-transferase enzyme of the Egyptian cattle tick R. annulatus (rRaGST), in order to confirm their ability to inhibit the parasitic essential detoxification enzyme glutathione S-transferase. Extraction with 70% ethanol of Hibiscus cannabinus (kenaf flowers), Punica granatum (red and white pomegranate peel), Musa acuminata (banana peel) (Musaceae), Medicago sativa (alfalfa seeds), Tamarindus indicus (seed) and Cuminum cyminum (cumin seed) were used to assess: (i) inhibitory capacities of rRaGST and (ii) their phenolic and flavonoid contents. Ethanol extraction of red pomegranate peel contained the highest content of phenolic compounds (29.95mg gallic acid/g dry tissue) compared to the other studied plant extracts. The highest inhibition activities of rRaGST were obtained with kenaf and red pomegranate peel (P. granatum) extracts with IC50 values of 0.123 and 0.136mg dry tissue/ml, respectively. Tannic acid was the more effective inhibitor of rRaGST with an IC50 value equal to 4.57μM compared to delphinidine-HCl (IC50=14.9±3.1μM). Gossypol had a weak inhibitory effect (IC50=43.7μM), and caffeic acid had almost no effect on tick GST activity. The IC50 values qualify ethacrynic acid as a potent inhibitor of rRaGST activity (IC50=0.034μM). Cibacron blue and hematin showed a considerable inhibition effect on rRaGST activity, and their IC50 values were 0.13μM and 7.5μM, respectively. The activity of rRaGST was highest for CDNB (30.2μmol/min/mg protein). The enzyme had also a peroxidatic activity (the specific activity equals 26.5μmol/min/mg protein). Both tannic acid and hematin inhibited rRaGST activity non-competitively with respect to GSH and

  17. Comparison of in vitro acaricidal effects of commercial preparations of cypermethrin and fenvalerate against Rhipicephalus (Boophilus) annulatus.

    PubMed

    Ravindran, Reghu; Ramankutty, Sunil Athalathil; Juliet, Sanis; Palayullaparambil, Adarsh Krishna Thumadath; Gopi, Jyothimol; Gopalan, Ajith Kumar Karapparambu; Nair, Suresh Narayanan; Ghosh, Srikanta

    2014-01-01

    Commercially available preparations of cypermethrin (Clinar and Ectomin) and fenvalerate (Flytik and Ticomax, 20% E.C) were compared for their acaricidal activity against Rhipicephalus (Boophilus) annulatus using adult immersion test. Adult tick mortality was higher with Ectomin compared to Clinar. Complete eclosion blocking was observed at all the tested concentrations with Ectomin while it was observed only at the highest concentration tested for Clinar. Compared to Flytik, adult tick mortality was higher with Ticomax at the tested concentrations. Complete blocking of hatching of laid ova was observed with Flytik at the highest concentration tested. At the manufacture recommended dosage of 200 ppm Ectomin elicited 93.37 per cent inhibition of fecundity, while it was 91.7 per cent for Clinar. For fenvalerate, the recommended concentration was 1200 ppm at which Ticomax showed 86 per cent and Flytik produced 80.05 per cent inhibition of fecundity respectively.

  18. Widespread Movement of Invasive Cattle Fever Ticks (Rhipicephalus microplus) in Southern Texas Leads to Shared Local Infestations on Cattle and Deer

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Rhipicephalus microplus is a highly-invasive tick that vectors cattle fever parasites (Babesia bovis and B. bigemina). In North America, ticks and Babesia are endemic in Mexico and ticks persist in the United States inside a narrow permanent quarantine zone (PQZ) along the Rio Grande. Th...

  19. Acaricide and Ivermectin resistance in a field population of Rhipicephalus microplus (Acari: Ixodidae) collected from Red Deer (Cervus elaphus) in the Mexican Tropics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Red deer (Cervus elaphus), taurine (Bos taurus) and zebuine (Bos indicus) breeds of cattle and their crosses, and the southern cattle fever tick (Rhipicephalus microplus) are non-native species that were introduced to Mexico through the livestock trade. Red deer raised in the Neotropics can die from...

  20. High resolution predictive mapping of Rhipicephalus microplus and R. annulatus in south Texas after vaccination with the anti-tick vaccine Gavac

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Conventional anti-tick vaccines based on the tick gut antigen Bm86 exist commercially (TickGARD (TM) and Gavac (R)) and could serve as an alternative to the use of acaricides to eradicate ticks, but their level of efficacy against R. microplus is too low for eradication if used alone. Therefore, the...

  1. Acaricidal activity of methanol extract of Acmella oleracea L. (Asteraceae) and spilanthol on Rhipicephalus microplus (Acari: Ixodidae) and Dermacentor nitens (Acari: Ixodidae).

    PubMed

    Cruz, Paula Barroso; Barbosa, Alan Franco; Zeringóta, Viviane; Melo, Diego; Novato, Tatiane; Fidelis, Queli Cristina; Fabri, Rodrigo Luiz; de Carvalho, Mário Geraldo; Oliveira Sabaa-Srur, Armando Ubirajara; Daemon, Erik; Monteiro, Caio Márcio Oliveira

    2016-09-15

    We evaluated the acaricidal activity of Acmella oleracea methanol extract and spilanthol on Rhipicephalus microplus and Dermacentor nitens. The extract was made through maceration with methanol. From this extract, a dichloromethane fraction with 99% spilanthol was obtained and tested on R. microplus larvae and engorged females and D. nitens larvae. For evaluation against larvae, the modified larval packet test was used, and both the methanol extract and dichloromethane fraction were tested at concentrations of 0.2-50mg/mL. The modified larval packet test was also used in the lethal time (LT) test, with the methanol extract at a concentration of 12.5mg/mL and the percentage mortality was assessed after 15, 30, 45, 60, 75, 90, 120min and 24h. The 50% lethal time calculation (LT50) was performed in this test. The engorged female test was performed with R. microplus only, at concentrations of 25-200mg/mL for methanol extract and 2.5-20.0mg/mL for spilanthol. The methanol extract caused 100% mortality of the R. microplus and D. nitens larvae at concentrations of 3.1 and 12.5mg/mL, respectively. Spilanthol resulted in 100% mortality of R. microplus larvae at concentration of 1.6mg/mL and of D. nitens at 12.5mg/mL. In the lethal time assay using the methanol extract, the mortality rate was 100% for R. microplus and D. nitens larvae after 120min and 24h, with LT50 values of 38 and 57min, respectively. In the test of females, the egg mass weight and the hatching percentage of the groups treated with concentrations equal to or higher than 50.0mg/mL of methanol extract were significantly reduced (p<0.05), while for spilanthol, the reduction of the egg mass weight and hatching percentage occurred from concentrations of 10.0mg/mL and 2.5mg/mL, respectively. Females treated with 200.0mg/mL of extract died before starting oviposition, resulting in 100% effectiveness, while the best efficacy for spilanthol was 92.9% at a concentration of 20.0mg/mL. Thus we conclude that the

  2. Mutation in the RmβAOR gene is associated with amitraz resistance in the cattle tick Rhipicephalus microplus

    PubMed Central

    Corley, Sean W.; Jonsson, Nicholas N.; Piper, Emily K.; Cutullé, Christian; Stear, Michael J.; Seddon, Jennifer M.

    2013-01-01

    We aimed to describe the evolution of resistance to amitraz in Rhipicephalus microplus in the field and to test the association between amitraz resistance and the frequency of a mutation in the β-adrenergic octopamine receptor gene (RmβAOR). We established six populations of Rhipicephalus microplus ticks in similar paddocks by the admixture of ticks from strains known to be susceptible and resistant to amitraz and synthetic pyrethroids. Each population was managed using one of three acaricide treatment regimes: always amitraz, always spinosad, or rotation between amitraz and spinosad. We used microsatellites to elucidate population structure over time, an SNP in the para-sodium channel gene previously demonstrated to confer resistance to synthetic pyrethroids to quantify changes in resistance to synthetic pyrethroids over time, and a nonsynonymous SNP in the RmβAOR, a gene that we proposed to confer resistance to amitraz, to determine whether selection with amitraz increased the frequency of this mutation. The study showed panmixia of the two strains and that selection of ticks with amitraz increased the frequency of the RmβAOR mutation while increasing the prevalence of amitraz-resistance. We conclude that polymorphisms in the RmβAOR gene are likely to confer resistance to amitraz. PMID:24082133

  3. Bacteriological analysis of saliva from partially or fully engorged female adult Rhipicephalus microplus by next-generation sequencing.

    PubMed

    Xiang, Liangliang; Poźniak, Błażej; Cheng, Tian-Yin

    2017-01-01

    Tick-borne diseases are a major epidemiological problem worldwide. The aim of this study was to investigate the bacterial composition of saliva obtained from engorged adult Rhipicephalus microplus females. Saliva samples collected from partially or fully engorged adult female ticks were analysed using an ultra-high-throughput Illumina HiSeq 2500 sequencing system. To elucidate the possible routes of bacterial transmission, the bacterial flora from whole ticks were also investigated. Proteobacteria, Firmicutes, and Actinobacteria were the predominant phyla in all samples, and Acinetobacter, Rickettsia, Escherichia and Coxiella were the major genera. Microbial diversity in saliva samples from partially engorged ticks was more complex than that of samples from fully engorged individuals. The comparison of saliva and whole-tick samples suggests that bacteria in saliva also colonize the tick's body. We believe that some bacterial genera, such as Dermacoccus, Achromia, SMB53, Sutterella, Providencia, Mycoplana, Oscillospira, and Agrobacterium, were found and reported in ticks for the first time. The Coxiella and Rickettsia detected in this study might be tick-borne pathogens, suggesting health risks associated with exposure to R. microplus in humans and animals. These findings may serve as the basis for developing strategies to control ticks and tick-borne diseases.

  4. Evidence of a tick RNAi pathway by comparative genomics and reverse genetics screen of targets with known loss-of-function phenotypes in Drosophila

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  5. Global change and integrated approach for sustainable cattle fever tick eradication in the United States of America

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ticks Rhipicephalus (Boophilus) annulatus and R. (B.) microplus, commonly known as cattle and southern cattle tick, respectively, impede the development and sustainability of livestock industries throughout the tropics and other world regions. They affect animal productivity and wellbeing direct...

  6. Challenges with the southern cattle fever tick in Puerto Rico: Then and now

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bovine babesiosis and anaplasmosis are deadly cattle diseases caused by microorganisms transmitted by the southern cattle fever tick (SCFT), Rhipicephalus (Boophilus) microplus, which is considered the most economically important ectoparasite of livestock worldwide. Humans brought animals infested w...

  7. Species distribution modelling for Rhipicephalus microplus (Acari: Ixodidae) in Benin, West Africa: comparing datasets and modelling algorithms.

    PubMed

    De Clercq, E M; Leta, S; Estrada-Peña, A; Madder, M; Adehan, S; Vanwambeke, S O

    2015-01-01

    Rhipicephalus microplus is one of the most widely distributed and economically important ticks, transmitting Babesia bigemina, B. bovis and Anaplasma marginale. It was recently introduced to West Africa on live animals originating from Brazil. Knowing the precise environmental suitability for the tick would allow veterinary health officials to draft vector control strategies for different regions of the country. To test the performance of modelling algorithms and different sets of environmental explanatory variables, species distribution models for this tick species in Benin were developed using generalized linear models, linear discriminant analysis and random forests. The training data for these models were a dataset containing reported absence or presence in 104 farms, randomly selected across Benin. These farms were sampled at the end of the rainy season, which corresponds with an annual peak in tick abundance. Two environmental datasets for the country of Benin were compared: one based on interpolated climate data (WorldClim) and one based on remotely sensed images (MODIS). The pixel size for both environmental datasets was 1 km. Highly suitable areas occurred mainly along the warmer and humid coast extending northwards to central Benin. The northern hot and drier areas were found to be unsuitable. The models developed and tested on data from the entire country were generally found to perform well, having an AUC value greater than 0.92. Although statistically significant, only small differences in accuracy measures were found between the modelling algorithms, or between the environmental datasets. The resulting risk maps differed nonetheless. Models based on interpolated climate suggested gradual variations in habitat suitability, while those based on remotely sensed data indicated a sharper contrast between suitable and unsuitable areas, and a patchy distribution of the suitable areas. Remotely sensed data yielded more spatial detail in the predictions. When

  8. First report of fluazuron resistance in Rhipicephalus microplus: a field tick population resistant to six classes of acaricides.

    PubMed

    Reck, José; Klafke, Guilherme Marcondes; Webster, Anelise; Dall'Agnol, Bruno; Scheffer, Ramon; Souza, Ugo Araújo; Corassini, Vivian Bamberg; Vargas, Rafael; dos Santos, Julsan Silveira; Martins, João Ricardo de Souza

    2014-03-17

    The control of the cattle tick Rhipicephalus microplus is based mainly on the use of chemical acaricides, which has contributed to the emerging problem of selection of resistant tick populations. Currently, there are six main classes of acaricides commercially available in Brazil to control cattle ticks, with fluazuron, a tick growth regulator with acaricidal properties, being the only active ingredient with no previous reports of resistance. Ticks (designated the Jaguar strain) were collected in a beef cattle ranch located at Rio Grande do Sul state, Southern Brazil, after a complaint of fluazuron treatment failure. To characterise the resistance of this strain against acaricides, larval tests were performed and showed that the Jaguar strain was resistant to all of the drugs tested: cypermethrin (resistance ratio, RR=31.242), chlorpyriphos (RR=103.926), fipronil (RR=4.441), amitraz (RR=11.907) and ivermectin (3.081). A field trial was conducted to evaluate the efficacy of fluazuron treatment in heifers that had been experimentally infested with the Jaguar or a susceptible strain. Between 14 and 28 days after treatment, the average efficacy in cattle experimentally infested with the susceptible strain was 96%, while for the Jaguar strain the efficacy was zero. Additionally, the Jaguar strain response to fluazuron was evaluated in vitro using a modified adult immersion test (AIT) and the artificial feeding assay (AFA). With the AIT, 50 ppm of fluazuron inhibited 99% of larvae hatching in the susceptible strain (POA) and less than 50% in the Jaguar strain. Results of the AFA showed a larval hatching rate of 67% at 2.5 ppm of fluazuron with the Jaguar strain; conversely, only 3% of larvae of the susceptible strain hatched at the same fluazuron concentration. The results showed here demonstrated the first case of fluazuron resistance in R. microplus and the first tick population resistant to six classes of acaricides in Brazil.

  9. Assessment of the acaricidal activity of carvacrol, (E)-cinnamaldehyde, trans-anethole, and linalool on larvae of Rhipicephalus microplus and Dermacentor nitens (Acari: Ixodidae).

    PubMed

    de Oliveira Souza Senra, Tatiane; Zeringóta, Viviane; de Oliveira Monteiro, Caio Márcio; Calmon, Fernanda; Maturano, Ralph; Gomes, Geovany Amorim; Faza, Aline; de Carvalho, Mario Geraldo; Daemon, Erik

    2013-04-01

    The acaricidal activity of carvacrol, (E)-cinnamaldehyde, trans-anethole, and linalool was studied on Rhipicephalus microplus and Dermacentor nitens larvae. All the substances were tested at concentrations of 2.5, 5.0, 10.0, 15.0, and 20.0 μl/ml, with 10 repetitions per treatment. The modified larval packet technique was employed in the tests and the mortality was evaluated after 24 h. In the groups treated with carvacrol, the lowest concentration (2.5 μl/ml) was sufficient to cause 100% death of the R. microplus and D. nitens larvae. The same concentration of (E)-cinnamaldehyde resulted in death of approximately 99% of the larvae of both tick species and reached 100% at the other concentrations. For trans-anethole, mortality rates above 90% of the R. microplus and D. nitens larvae were only observed starting at the concentration of 15.0 μl/ml and reached 100% at the highest concentration (20.0 μl/ml). Finally, the mortality rates of the groups treated with linalool were low, only reaching 8.4 and 14.5% at the highest concentration (20.0 μl/ml) for larvae of D. nitens and R. microplus, respectively. These results show that carvacrol, (E)-cinnamaldehyde, and trans-anethole have acaricidal activity, particularly carvacrol and (E)-cinnamaldehyde, both of which resulted in high mortality rates for the larvae of these two tick species even at the lowest concentration.

  10. Acaricidal activity of the organic extracts of thirteen South African plants against Rhipicephalus (Boophilus) decoloratus (Acari: Ixodidae).

    PubMed

    Fouche, Gerda; Ramafuthula, Mary; Maselela, Vusi; Mokoena, Moses; Senabe, Jeremiah; Leboho, Tlabo; Sakong, Bellonah M; Adenubi, Olubukola T; Eloff, Jacobus N; Wellington, Kevin W

    2016-07-15

    The African blue tick, Rhipicephalus (Boophilus) decoloratus, is a common tick species found in South Africa and affects cattle production as well as vectoring pathogens in regions of Africa and Asia. In an attempt to develop a non-toxic, lower cost and environmentally friendly tick control method, twenty-six plant extracts were prepared from thirteen plant species using 99.5% acetone and 99% ethanol. The adapted Shaw Larval Immersion Test (SLIT) was used to test the efficacy of the extracts. A 1% solution of each of the plant extracts was prepared for efficacy testing and the ethanol extracts were found to have better acaricidal activity than the acetone extracts. The ethanol extract from the leaves and flowers of Calpurnia aurea had the best activity [corrected mortality (CM)=82.9%] which was followed by the stem extract of Cissus quadrangularis (CM=80.4%). The plant species were screened against Vero cells and were found to have low toxicity. From this study it is apparent that there is potential for the development of botanicals as natural acaricides against R. (B.) decoloratus.

  11. Acaricidal activity of petroleum ether extract of leaves of Tetrastigma leucostaphylum (Dennst.) Alston against Rhipicephalus (Boophilus) annulatus.

    PubMed

    Krishna, T P Adarsh; Krishna, T P Ajeesh; Chithra, N D; Deepa, P E; Darsana, U; Sreelekha, K P; Juliet, Sanis; Nair, Suresh N; Ravindran, Reghu; Kumar, K G Ajith; Ghosh, Srikanta

    2014-01-01

    The acaricidal activity of the petroleum ether extract of leaves of Tetrastigma leucostaphylum (Dennst.) Alston (family: Vitaceae) against Rhipicephalus (Boophilus) annulatus was assessed using adult immersion test (AIT). The per cent of adult mortality, inhibition of fecundity, and blocking of hatching of eggs were studied at different concentrations. The extract at 10% concentration showed 88.96% inhibition of fecundity, 58.32% of adult tick mortality, and 50% inhibition of hatching. Peak mortality rate was observed after day 5 of treatment. Mortality of engorged female ticks, inhibition of fecundity, and hatching of eggs were concentration dependent. The LC50 value of the extract against R. (B.) annulatus was 10.46%. The HPTLC profiling of the petroleum ether extract revealed the presence of at least seven polyvalent components. In the petroleum ether extract, nicotine was identified as one of the components up to a concentration of 5.4%. However, nicotine did not reveal any acaricidal activity up to 20000 ppm (2%). Coconut oil, used as diluent for dissolving the extract, did not reveal any acaricidal effects. The results are indicative of the involvement of synergistic or additive action of the bioactive components in the tick mortality and inhibition of the oviposition.

  12. Tick salivary acetylcholinesterase: A probable immunomodulator of host-parasite interactions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini), is the most economically important ectoparasite affecting cattle in the world. Although eradicated from the United States, R. microplus and R. annulatus (Say) continue to threaten U.S. cattle producers despite maintenance of...

  13. Mitigation of Resistance Through Mixtures of Traditional Pesticides, Anti-tick Vaccines, and New Acaricides Developed by the Pharmaceutical Industry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Over the past 70 years, the southern cattle tick, Rhipicephalus (Boophilus) microplus, has developed resistance to every acaricide available for its control. Recently, populations of R. microplus have evolved simultaneous resistance to multiple classes of acaricides. These multi-resistant population...

  14. Acaricidal effect of Pelargonium roseum and Eucalyptus globulus essential oils against adult stage of Rhipicephalus (Boophilus) annulatus in vitro.

    PubMed

    Pirali-Kheirabadi, Khodadad; Razzaghi-Abyaneh, Mehdi; Halajian, Ali

    2009-06-10

    In a laboratory trial, in west-central Iran, the acaricidal effects of the essential oils (EOs) prepared from two medicinal plants, i.e. Pelargonium roseum and Eucalyptus globulus on the adult stage of Rhipicephalus (Boophilus) annulatus were evaluated. For this purpose, the engorged females of R. (B) annulatus were exposed to two-fold serial dilutions of oils (0.31-5.0%) using a "dipping method" in vitro. The engorged ticks were immersed in different plant dilutions (eight per dilution) for 1min then each replicate was incubated in separate petri dishes at 26 degrees C and 80% relative humidity. The mortality rate for adult ticks exposed to different dilutions of P. roseum and E. globulus EO's showed a dose-dependent decrease. It was however significant only for the 2.5% and 5.0% dilutions of P. roseum EO, when compared to the non-treated control (P<0.05). The mass of produced eggs in adult female ticks exposed to both P. roseum and E. globulus EOs had decreased dose-dependently. It was significant for only 2.5% and 5.0% dilutions of P. roseum EO, comparing the non-treated control (P<0.05). The highest decrease in egg laying was reported for ticks treated with 5% dilutions of P. roseum (87.5%) and E. globosus (25%) (P<0.05). This is the first report that details the acaricidal activity of EO's obtained from P. roseum and E. globosus against R. (B) annulatus. The results show that both plants, particularly P. Roseum can be considered as potential candidates for biocontrol of R. (B) annulatus in the field.

  15. Lavandula angustifolia essential oil as a novel and promising natural candidate for tick (Rhipicephalus (Boophilus) annulatus) control.

    PubMed

    Pirali-Kheirabadi, Khodadad; Teixeira da Silva, Jaime A

    2010-10-01

    Lavandula angustifolia is a well known herbal medicine with a variety of useful properties, including its acaricidal effect. This experiment was carried out to study the bioacaricidal activity of L. angustifolia essential oil (EO) against engorged Rhipicephalus (Boophilus) annulatus (Acari; Ixodidae) females. For this purpose six serial concentrations (0.5, 1.0, 2.0, 4.0, 6.0 and 8.0% w/v) of L. angustifolia EO were used. There was considerable mortality in concentrations more than 4.0% although there were no differences between 6.0 and 8.0% in all measured criteria. The mortality rate 24 h after inoculation was 73.26 and 100% in groups treated with 4.0 and 8.0% EO, respectively. Lavender EO also reduced tick egg weight in a concentration-dependent manner. The amount of eggs produced varied from 0.12 g (at 0.5% EO) to 0.00 g (at 8.0% EO) but did not differ statistically from the control. L. angustifolia EO caused 100% failure in egg laying at 6.0 and 8.0% whereas this value in the control group was zero. A positive correlation between L. angustifolia EO concentration and tick control, assessed by relative mortality rate and egg-laying weight, was observed by the EO LC/EC(50), which, when calculated using the Probit test, was 2.76-fold higher than the control. Lavender is a promising acaricidal against R. (B.) annulatus in vitro.

  16. Ehrlichia minasensis sp. nov., a new species within the genus Ehrlichia isolated from the tick Rhipicephalus microplus.

    PubMed

    Cabezas-Cruz, Alejandro; Zweygarth, Erich; Vancová, Marie; Broniszewska, Marzena; Grubhoffer, Libor; Passos, Lygia Maria Friche; Ribeiro, Múcio Flávio Barbosa; Alberdi, Pilar; de la Fuente, José

    2016-01-12

    Recently, we obtained a rickettsial isolate (Ehrlichia sp. UFMG-EV) from the hemolymph of Rhipicephalus microplus engorged tick females. On the basis of maximum likelihood phylogenetic analysis using 16S rRNA, groEL, dsb, gltA and trp36 nucleotide sequences we showed that Ehrlichia sp. UFMG-EV belongs to the α-Proteobacteria, family Anaplasmataceae, genus Ehrlichia. Ehrlichia sp. UFMG-EV is a sister taxa of E. canis with 16S rRNA, groEL, dsb, gltA and trp36 nucleotide sequence similarity of 98.3%, 97.2%, 94.7%, 94.3% and 49.1%, respectively. Ehrlichia sp. UFMG-EV has been maintained in the laboratory by continuous passage in the IDE8 tick cell line where the ultrastructure was characterized using electron microscopy and it resembles that of E. canis, E. muris and E. chaffensis, but not E. ruminantium and E. ewingii. We propose the name Ehrlichia minasensis sp. nov. for this bacterium to acknowledge the place from where it was isolate for first time, Minas Gerais, Brazil. The type strain of Ehrlichia minasensis sp. nov. is strain Ehrlichia sp. UFMG-EV (= DSM 100393, = TCB-TBB-0018).

  17. Syringe test (modified larval immersion test): a new bioassay for testing acaricidal activity of plant extracts against Rhipicephalus microplus.

    PubMed

    Sindhu, Zia-ud-Din; Jonsson, Nicholas N; Iqbal, Zafar

    2012-09-10

    We report a new bioassay "syringe test" (modified larval immersion test) for in vitro evaluation of acaricidal activity of crude plant extracts. Prepared syringes, containing eggs of tick, were incubated until 14 d after hatching of eggs, when the bioassay was performed on the larvae. Lethal concentrations for 50% of larvae (LC(50)), LC(90) and LC(99) values were calculated for each tested product. 95% confidence intervals for LC(50) were very narrow, indicating a high degree of repeatability for the new bioassay on larvae of R. microplus. Bioassays were applied to six crude aqueous-methanol extracts from five plants (Acacia nilotica, Buxus papillosa, Fumaria parviflora, Juniperus excelsa, and Operculina turpethum), of which three showed discernible effects. Twenty-four hours post exposure, LC(99) values were 11.9% (w/v) for F. parviflora, 20.8% (w/v) and 29.2% (w/v) for B. papillosa and A. nilotica, respectively. After six days of exposure these values were; 9.1% (w/v), 9.2% (w/v) and 15.5 (w/v) for F. parviflora, A. nilotica and B. papillosa, respectively.

  18. Acaricidal activity of essential oils from Lippia alba genotypes and its major components carvone, limonene, and citral against Rhipicephalus microplus.

    PubMed

    Peixoto, Magna Galvão; Costa-Júnior, Livio Martins; Blank, Arie Fitzgerald; Lima, Aldilene da Silva; Menezes, Thays Saynara Alves; Santos, Darlisson de Alexandria; Alves, Péricles Barreto; Cavalcanti, Sócrates Cabral de Holanda; Bacci, Leandro; Arrigoni-Blank, Maria de Fátima

    2015-05-30

    The goal of the present study was to evaluate the acaricidal potential of Lippia alba essential oil, citral chemotypes (LA-10 and LA-44 genotypes) and carvone chemotypes (LA-13 and LA-57 genotypes), as well as purified citral and enantiomers of carvone and limonene. Efficacy against Rhipicephalus microplus was assessed by the larval packet and the engorged female immersion tests. Citral chemotypes had greater larvicidal activity than carvone chemotypes, and this was further supported by larvicidal and adulticidal activity of purified citral with LC50 values of 7.0 and 29.8 mg/mL, respectively. While purified enantiomers of carvone exhibited greater larvicidal activity than those of limonene, enantioselectivity of limonene was observed with R-(+) displaying significantly higher efficacy (LC50 of 31.2mg/mL) than S-(-) (LC50 of 54.5mg/mL). The essential oils and purified compounds were much less toxic toward engorged adult females, with the exception of citral, and this may be due to limited cuticular penetration.

  19. Chemical composition and acaricidal activity of essential oil from Lippia sidoides on larvae of Dermacentor nitens (Acari: Ixodidae) and larvae and engorged females of Rhipicephalus microplus (Acari: Ixodidae).

    PubMed

    Gomes, Geovany Amorim; Monteiro, Caio Márcio de Oliveira; Senra, Tatiane de Oliveira Souza; Zeringota, Viviane; Calmon, Fernanda; Matos, Renata da Silva; Daemon, Erik; Gois, Roberto Wagner da Silva; Santiago, Gilvandete Maria Pinheiro; de Carvalho, Mario Geraldo

    2012-12-01

    The aim of this work was to identify the compounds and to investigate the acaricidal activity of the essential oil from the leaves of Lippia sidoides on Rhipicephalus microplus and Dermacentor nitens. The oil was obtained by hydrodistillation and analyzed by gas chromatography (GC/FID) and gas chromatography/mass spectrometry. In total, 15 compounds comprising 99.97 % of the total peak area were identified. The main constituent of the essential oil was thymol (67.60 %). The acaricidal activity was assessed by the modified larval packet test, with oil concentrations of 2.5, 5.0, 10.0, 15.0, and 20.0 μl/ml, and by the female immersion test with concentrations of 10.0, 20.0, 40.0, 60.0, and 80.0 μl/ml. The mortality of the R. microplus and D. nitens larvae was greater than 95 % starting at concentrations of 10.0 and 20.0 μl/ml, respectively. In the test with the engorged females, the L. sidoides essential oil starting at a concentration of 40.0 μl/ml caused a significant reduction (p < 0.05) in the values of the egg mass weight and egg production index. The viability of the eggs was affected in all the treated groups, with significantly lower hatching rates (p < 0.05) in relation to the control group. The control percentages at concentrations of 10.0, 20.0, and 30.0 μl/ml were 54, 57, and 72 %, and reached 100 % at the highest two concentrations (60.0 and 80.0 μl/ml). Therefore, it can be concluded that the essential oil from the leaves of L. sidoides has acaricidal activity on R. microplus and D. nitens.

  20. Protective action of Tagetes minuta (Asteraceae) essential oil in the control of Rhipicephalus microplus (Canestrini, 1887) (Acari: Ixodidae) in a cattle pen trial.

    PubMed

    Andreotti, Renato; Garcia, Marcos Valério; Cunha, Rodrigo Casquero; Barros, Jacqueline Cavalcante

    2013-10-18

    The Rhipicephalus microplus tick is globally regarded as the most economically important ectoparasite of livestock, and the evolution of resistance to commercial acaricides among cattle tick populations is of great concern. The essential oil derived from Tagetes minuta may be efficacious against cattle tick infestation, and the results of a cattle pen trial using this essential oil for the control of ticks are reported here. The chemical composition of the essential oil was determined by GC-MS and NMR spectroscopy analyses, which revealed the presence of four major components in the essential oil. These components represent more than 70% of the essential oil: limonene (6.96%), β-ocimene (5.11%), dihydrotagetone (54.10%) and tagetone (6.73%). The results of the cattle pen trial indicated significant differences among the average values of the analyzed biological parameters, including the number of ticks, the average weight of the ticks, the average egg weight per engorged female and larval viability. Treatment with the T. minuta essential oil prepared in this study promoted significant effects on all biological indicators analyzed. Based on the biological indicators, the essential oil showed 99.98% efficacy compared to the control group when used at a 20% concentration. The results obtained in this study suggest that the T. minuta essential oil is a potential R. microplus tick control agent and may be used to mitigate the economic losses caused by tick infestation.

  1. Spectroscopic evaluation of thymol dissolved by different methods and influence on acaricidal activity against larvae of Rhipicephalus microplus (Acari: Ixodidae).

    PubMed

    Daemon, Erik; Monteiro, Caio Márcio Oliveira; Maturano, Ralph; Senra, Tatiane Oliveira Souza; Calmon, Fernanda; Faza, Aline; de Azevedo Prata, Márcia Cristina; Georgopoulos, Stéfanos Leite; de Oliveira, Luiz Fernando Cappa

    2012-11-01

    The acaricidal activity of three thymol formulations was investigated at five concentrations (1.25, 2.5, 5.0, 7.5, and 10.0 mg/ml) on Rhipicephalus microplus larvae, and the behavior of its solubility in these formulations was analyzed. The thymol was dissolved in distilled water plus 1 % dimethylsulfoxide as adjuvant under two heating regimes (water bath in formulation 1 and hot plate in formulation 2) as well as without heating in 50 % ethanol and 50 % water (v/v). The acaricidal activity was assessed by the modified larval packet test, and the solubilization behavior was investigated by ultraviolet-visible spectroscopy, based on the Beer-Lambert law. With formulations 1 and 2, the mortality was greater than 95 % starting at the thymol concentrations of 5.0 and 7.5 mg/ml, respectively, while with formulation 3, this mortality level was reached starting at a concentration of 2.5 mg/ml, showing that the addition of ethanol in the solution enhanced the acaricidal action of thymol. This result was supported by the LC 90 values, which were 3.3, 2.4, and 1.6 mg/ml of thymol for formulations 1, 2, and 3, respectively. This result is related to the better solubility of thymol in the hydroethanolic formulation, since the spectroscopic analysis revealed that the thymol dissolved more completely in this formulation. This fact was evident once the R (2) obtained from the linear regression analysis of the relation absorbance × concentration of the formulations 1, 2, and 3 approached the optimal value (R (2) = 1) in the following sequence: 1, 2, and 3 (0.717, 0.901, and 0.968, respectively).

  2. Prevalence and genetic characterization of Anaplasma marginale in zebu cattle (Bos indicus) and their ticks (Amblyomma variegatum, Rhipicephalus microplus) from Madagascar.

    PubMed

    Pothmann, Daniela; Poppert, Sven; Rakotozandrindrainy, Raphael; Hogan, Benedikt; Mastropaolo, Mariano; Thiel, Claudia; Silaghi, Cornelia

    2016-10-01

    Tick-borne bovine anaplasmosis, caused by the obligate intracellular pathogen Anaplasma marginale (Rickettsiales: Anaplasmataceae), is a major constraint to cattle production in tropical and subtropical regions. From Madagascar, clinical cases were published but data based on molecular methods regarding the prevalence and genetic diversity of this pathogen on the island are lacking. The aims of this study were to investigate (1) the prevalence of A. marginale in Malagasy zebu cattle (Bos indicus) and their ticks with a species-specific real-time PCR, (2) the genetic diversity of A. marginale based on tandem repeats and microsatellites of the msp1α gene, and (3) the phylogenetic relationship between A. marginale isolates from Madagascar and strains found worldwide. Two hundred fourteen blood samples and 1822 ticks from 214 zebu cattle were collected. Rhipicephalus (R) microplus (40.2%) and Amblyomma (A) variegatum (59.8%) were identified on the cattle. A. marginale DNA was found in 89.7% of the examined zebu cattle and in 62.3% of the examined ticks. The tandem repeat and microsatellite analyses of the mspa1 gene showed high genetic diversity among the isolates between and within the different regions and high infection potential. Eighteen of the 25 tandem repeats identified have not been described before. Phylogenetic analysis revealed clustering of A. marginale strains from Madagascar with South Africa, America and Israel. A common ancestor may originate from South Africa and may have evolved due to phylogeographic characteristics or by a history of cattle movement. Its high prevalence in cattle and ticks, together with a low number of clinical manifestations and a high genetic heterogeneity among the investigated strains, confirms endemic stability of A. marginale in cattle from Madagascar.

  3. Evidence for the role of white-tailed deer(Artiodactyla:Cervidae)in the epidemiology of cattle ticks and southern cattle ticks (Acari:Ixodidae)in reinfestations along the Texas/Mexico border in South Texas-A review and update

    Technology Transfer Automated Retrieval System (TEKTRAN)

    From 1907 when the fever tick eradication campaign began until 1933 the tick eradication methods of dipping cattle in an acaricide or "pasture vacation" were enormously successful in eradicating southern cattle ticks [SCT, Rhipicephalus (Boophilus) microplus (Canestrini)], until failures began to oc...

  4. Identification, characterization and use of two tick promoters for construction of a dual luciferase reporter vector

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Dual luciferase reporter systems are valuable tools for functional genomic studies, but have not previously been developed for use in tick cell culture. We evaluated expression of available luciferase constructs in tick cell cultures derived from Rhipicephalus (Boophilus) microplus, an important vec...

  5. Preliminary findings of a molecular survey for the presence of B. bovis and B. bigemina in cattle fever ticks and white-tailed deer from south Texas

    Technology Transfer Automated Retrieval System (TEKTRAN)

    White-tailed deer are an alternative host for Rhipicephalus (Boophilus) microplus and R. (B.) annulatus, collectively referred to as cattle fever ticks. Dense white-tailed deer populations in south Texas complicate efforts by the National Cattle Fever Tick Eradication Program to keep the U.S. free o...

  6. Integrated strategy for sustainable cattle fever tick eradication in U.S.A. is required to mitigate the impact of global change

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ticks Rhipicephalus (Boophilus) annulatus and R.(B.) microplus, commonly known as cattle and southern cattle tick, respectively, impede the development and sustainability of livestock industries throughout tropical and other world regions. They affect animal productivity and wellbeing directly t...

  7. Research update on the potential use of anti-tick vaccines to support the USDA Cattle Fever Tick Eradication Program

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The USDA APHIS VS Cattle Fever Tick Eradication program (CFTEP) was established in 1906 to eliminate populations of the cattle fever ticks (CFT) Rhipicephalus (Boophilus) annulatus and R. microplus as a way to eradicate bovine babesiosis from the continental United States, which is a high-impact dis...

  8. Integration of ecologically-based approaches to re-eradicate cattle fever ticks from the U.S.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Here we summarize highlights of research conducted as part of a NIFA-AFRI funded grant. Cattle fever ticks, Rhipicephalus (Boophilus) microplus and R. (B.) annulatus, have been found on white-tailed deer (Odocoileus virginianus) complicating eradication efforts of the USDA’s Cattle Fever Tick Eradic...

  9. Shift in rules of engagement for eradication of cattle fever ticks in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    With the exception of a systematic quarantine zone in south Texas along the border with Mexico, cattle fever ticks (CFT), i.e. Rhipicephalus (Boophilus) microplus and R. (B.) annulatus, were officially eradicated from the U.S. in 1943. Because of their roles as vectors of bovine babesiosis, the re-e...

  10. Cattle Fever Ticks in the U.S.: Back to 1906?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Keeping cattle fever ticks (CFT), Rhipicephalus (Boophilus) microplus and R. annulatus, eradicated from the United States and thus keeping the national cattle herd free of bovine babesiosis is a current and critical agricultural biosecurity issue of national relevance. Also known as “Texas fever”, ...

  11. Research project for integrated control of the southern cattle fever tick in Puerto Rico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Puerto Rico (PR) is infested with the southern cattle fever tick (SCFT), Rhipicephalus (Boophilus) microplus, which is considered the most economically important external parasite of livestock worldwide. A research coalition involving the livestock industry in PR, the PR Department of Agriculture (...

  12. Chemical composition and acaricidal activity of the essential oil of Baccharis dracunculifolia De Candole (1836) and its constituents nerolidol and limonene on larvae and engorged females of Rhipicephalus microplus (Acari: Ixodidae).

    PubMed

    de Assis Lage, Tiago Coelho; Montanari, Ricardo Marques; Fernandes, Sergio Antonio; de Oliveira Monteiro, Caio Márcio; de Oliveira Souza Senra, Tatiane; Zeringota, Viviane; da Silva Matos, Renata; Daemon, Erik

    2015-01-01

    Baccharis dracunculifolia DC (common name "alecrim-do-campo" in Brazil) is a plant with widespread distribution in South America that is the botanical origin of green propolis. The aim of this study was to evaluate the chemical composition and acaricidal activity of the essential oil of B. dracunculifolia and its constituents nerolidol and limonene on unengorged larvae and engorged females of Rhipicephalus microplus (Acari: Ixodidae). The essential oil yield was 0.8% of dry mass and the major constituents were nerolidol (22.3%), germacrene D (7.2%), limonene (6.9%), β-pinene (6.7) and bicyclogermacrene (6.5%). The acaricidal activity of the essential oil and the pure compounds nerolidol and (R)-(+)-limonene were assessed in the laboratory through the modified larval packet test (LPT) and the female immersion test (FIT). In the LPT, the essential oil and nerolidol were both active, causing more than 90% mortality at concentrations from 15.0 and 10.0 mg mL(-1), respectively, whereas (R)-(+)-limonene was not active. In the FIT, the oil and nerolidol caused reduction in the quantity and quality of eggs produced, with control percentages of 96.3% and 90.3% at concentrations of 60.0 and 50.0 mg mL(-1), respectively. It can be concluded that the essential oil obtained from the aerial parts of B. dracunculifolia and its major component nerolidol have high activity on R. microplus larvae and engorged females.

  13. Effect of Calea serrata Less. n-hexane extract on acetylcholinesterase of larvae ticks and brain Wistar rats.

    PubMed

    Ribeiro, Vera Lucia Sardá; Vanzella, Cláudia; Moysés, Felipe dos Santos; Santos, Jaqueline Campiol Dos; Martins, João Ricardo Souza; von Poser, Gilsane Lino; Siqueira, Ionara Rodrigues

    2012-10-26

    Acetylcholinesterase (AChE), an enzyme that hydrolyses acetylcholine (ACh) at cholinergic synapses, is a target for pesticides and its inhibition by organophosphates leads to paralysis and death of arthropods. It has been demonstrated that the n-hexane extract of Calea serrata had acaricidal activity against larvae of Rhipicephalus (Boophilus) microplus and Rhipicephalus sanguineus. The aim of the present study was to understand the mechanism of the acaricidal action of C. serrata n-hexane extract are specifically to investigate the in vitro anticholinesterase activity on larvae of R. microplus and in brain structures of male Wistar rats. The n-hexane extract significantly inhibited in vitro acetylcholinesterase activity in R. microplus larvae and rat brain structures. The results confirm that inhibition of acetylcholinesterase is a possible mechanism of action of hexane extract at C. serrata.

  14. Detection of Borrelia theileri in Rhipicephalus geigyi from Mali.

    PubMed

    McCoy, Brandi N; Maïga, Ousmane; Schwan, Tom G

    2014-06-01

    Disease burden within cattle is a concern around the world. Bovine borreliosis, one such disease, is caused by the spirochete Borrelia theileri transmitted by the bite of an infected Rhipicephalus (Boophilus) species tick. A number of species within the genus are capable of transmitting the agent and are found on multiple continents. Cattle in the West African nation of Mali are infested with four species of Rhipicephalus ticks of the subgenus Boophilus: Rhipicephalus annulatus, Rhipicephalus microplus, Rhipicephalus decoloratus, and Rhipicephalus geigyi. To date, no reports of B. theileri within Mali have been documented. We tested 184 Rhipicephalus spp. ticks by PCR that were removed from cattle at a market near Bamako, Mali. One tick, R. geigyi, was positive for B. theileri, which confirmed the presence of this spirochete in Mali.

  15. Ixodid ticks on domestic animals in Samut Prakan Province, Thailand.

    PubMed

    Nithikathkul, C; Polseela, P; Changsap, B; Leemingsawat, S

    2002-01-01

    The prevalence of tick-harboring domestic animals, tick density, and the species of ticks were studied throughout the year 2000, in Muang Samut Prakan, Bang Phli and Phra Pradaeng districts of Samut Prakan Province. The animals examined were Canis lupus familiaris (450), Bos indicus (cross-bred) (189), Bos taurus (30), Bubalus bubalis (171) and Sus scrofa domestica (450). The total number of collected ticks was 1,491. The pigs did not harbor ticks. The stages of ticks collected were larvae, nymphs and adults. The prevalence rates of tick-harboring were 46% (Canis lupus familiaris), 42.86% (Bos indicus, cross-bred), 33.33% (Bos taurus) and 9.35% (Bubalus bubalis). The tick densities were 2.22 (Bos indicus, cross-bred), 2.16 (Canis lupus familiaris), 1.16 (Bos taurus) and 0.36 (Bubalus bubalis). Only 2 species of ixodid ticks, Boophilus microplus and Rhipicephalus sanguineus, were found. R. sanguineus was the dominant species of tick. The percentage of R. sanguineus was 65.2% and B. microplus was 34.8%. In Muang district, R. sanguineus was the dominant species in C. lupus familiaris; in Bang Phli district, B. microplus was the dominant species in Bos indicus (cross-bred). The density of B. microplus was high in the summer season; the density of R. sanguineus was high in the winter season. The number of ticks depended on the geographic location, animal host and season.

  16. Cattle tick-associated bacteria exert anti-biofilm and anti-Tritrichomonas foetus activities.

    PubMed

    Zimmer, K R; Seixas, A; Conceição, J M; Zvoboda, D A; Barros, M P; Tasca, T; Macedo, A J; Termignoni, C

    2013-05-31

    Research on microbiota in cattle tick and the evaluation of its activity against other microorganisms can contribute to identify new molecules potentially useful to control infections caused by bacteria and protozoa. Biofilms pose increasing problems worldwide, mainly due to their resistance to antimicrobial therapies and host immune response. In this study we investigate the ability Rhipicephalus (Boophilus) microplus-associated bacteria may exhibit to produce anti-biofilm and trichomonicidal compounds. Gut, ovary, salivary glands, and Gené organ were collected from engorged R. microplus female. Homogenates of each tissue were inoculated onto 15 distinct culture media. Anti-biofilm and trichomonicidal activities were analyzed by culturing each bacterium isolated in a liquid medium. Results showed that R. microplus cattle tick microflora varies for different tissues. Bacteria belonging to different genera (Aeromonas, Bacillus, Brevibacillus, Castelaniella, Comamonas, Kocuria, and Microbacterium) were identified. Interestingly, all bacterial species found displayed pronounced activity against Staphylococcus epidermidis and Pseudomonas aeruginosa biofilms, and also against the cattle pathogen Tritrichomonas foetus, confirming the hypothesis that cattle tick could be a source of bacteria active against pathogens. This is the first study showing that bacteria isolated from a tick exert anti-biofilm and trichomonicidal activities.

  17. Effect of attractant sex pheromone on immature larval stages of ixodid tick species.

    PubMed

    Ranju, R S; Latha, Bhaskaran Ravi; Leela, V; Basith, S Abdul

    2012-10-01

    Attractant sex pheromone (ASP) 2,6-dichloro phenol was used in the current study to evaluate the percentage attraction and the behavioural responses of the five ixodid tick species namely Rhipicephalus sanguineus, Rhipicephalus microplus (Boophilus microplus), Haemaphysalis bispinosa, Rhipicephalus haemaphysaloides and Hyalomma marginatum using petridish bioassay. Two concentrations of 2,6-DCP (0.1 M and 0.05 M) was used for the larval stages of all five ixodid tick species of which 0.1 M concentration was found to have maximum attraction. Trials with 0.1 M ASP revealed highest per cent of attraction in R. sanguineus larvae (71 %) followed by H. bispinosa (55 %) and R. microplus (55 %). With 0.1 M ASP R. haemaphysaloides and H. marginatum showed least attraction (39 % each). However the per cent of attraction of R. haemaphysaloides was higher (46 %) with 0.05 M ASP. Statistical analysis revealed a highly significant difference in per cent of attraction between the different tick larvae using 0.05 and 0.1 M ASP. The larvae also exhibited behavioural responses such as feeding, probing, resting and questing posture.

  18. Interaction of plant essential oil terpenoids with the southern cattle tick tyramine receptor: A potential biopesticide target.

    PubMed

    Gross, Aaron D; Temeyer, Kevin B; Day, Tim A; Pérez de León, Adalberto A; Kimber, Michael J; Coats, Joel R

    2017-02-01

    An outbreak of the southern cattle tick, Rhipicephalus (Boophilus) microplus, (Canestrini), in the United States would have devastating consequences on the cattle industry. Tick populations have developed resistance to current acaricides, highlighting the need to identify new biochemical targets along with new chemistry. Furthermore, acaricide resistance could further hamper control of tick populations during an outbreak. Botanically-based compounds may provide a safe alternative for efficacious control of the southern cattle tick. We have developed a heterologous expression system that stably expresses the cattle tick's tyramine receptor with a G-protein chimera, producing a system that is amenable to high-throughput screening. Screening an in-house terpenoid library, at two screening concentrations (10 μM and 100 μM), has identified four terpenoids (piperonyl alcohol, 1,4-cineole, carvacrol and isoeugenol) that we believe are positive modulators of the southern cattle tick's tyramine receptor.

  19. Chemical identification of Tagetes minuta Linnaeus (Asteraceae) essential oil and its acaricidal effect on ticks.

    PubMed

    Garcia, Marcos Valério; Matias, Jaqueline; Barros, Jacqueline Cavalcante; de Lima, Dênis Pires; Lopes, Rosângela da Silva; Andreotti, Renato

    2012-01-01

    The control of tick species that affect animal production is vital for the economic welfare of the cattle industry. This study focused on testing the acaricidal activity of the essential oil from the leaves and stems of Tagetes minuta against several Brazilian tick species, including Rhipicephalus (Boophilus) microplus, Rhipicephalus sanguineus, Amblyomma cajennense and Argas miniatus. The chemical composition of the essential oil was determined by chromatography and spectroscopy analyses, which revealed the presence of monoterpenes. The adult immersion test (AIT) and the larval packet test (LPT) were used to evaluate the efficacy of T. minuta essential oil in tick management at concentrations of 2.5, 5, 10, 20 and 40%. The results demonstrated that the T. minuta essential oil had over 95% efficacy against four species of ticks at a concentration of 20%. These results suggest that the essential oil of T. minuta could be used as an environmentally friendly acaricide.

  20. Molecular survey and sequence analysis of Anaplasma spp. in cattle and ticks in a Malaysian farm.

    PubMed

    Tay, S T; Koh, F X; Kho, K L; Ong, B L

    2014-12-01

    This study was conducted to determine the occurrence of Anaplasma spp. in the blood samples of cattle, goats, deer and ticks in a Malaysian farm. Using polymerase chain reaction (PCR) and sequencing approach, Anaplasma spp. was detected from 81(84.4%) of 96 cattle blood samples. All blood samples from 23 goats and 22 deer tested were negative. Based on the analysis of the Anaplasma partial 16S ribosomal RNA gene, four sequence types (genotypes 1 to 4) were identified in this study. Genotypes 1-3 showed high sequence similarity to those of Anaplasma platys/ Anaplasma phagocytophilum, whilst genotype 4 was identical to those of Anaplasma marginale/ Anaplasma centrale/ Anaplasma ovis. Anaplasma DNA was detected from six (5.5%) of 109 ticks which were identified as Rhipicephalus (formely known as Boophilus) microplus ticks collected from the cattle. This study reported for the first time the detection of four Anaplasma sequence types circulating in the cattle population in a farm in Malaysia. The detection of Anaplasma DNA in R. microplus ticks in this study provides evidence that the ticks are one of the potential vectors for transmission of anaplasmosis in the cattle.

  1. Evaluation of green synthesized silver nanoparticles against parasites.

    PubMed

    Marimuthu, Sampath; Rahuman, Abdul Abdul; Rajakumar, Govindasamy; Santhoshkumar, Thirunavukkarasu; Kirthi, Arivarasan Vishnu; Jayaseelan, Chidambaram; Bagavan, Asokan; Zahir, Abdul Abduz; Elango, Gandhi; Kamaraj, Chinnaperumal

    2011-06-01

    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 (LC(50) = 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.

  2. Surveys for ectoparasites on wildlife associated with Amblyomma variegatum (Acari: Ixodidae)-infested livestock in St. Croix, U.S. Virgin Islands.

    PubMed

    Corn, Joseph L; Berger, Patrick; Mertins, James W

    2009-11-01

    Surveys in 2001, 2005, and 2006 attempted to determine the role of wildlife in maintenance and dissemination of the tropical bont tick, Amblyomma variegatum (F.) (Acari: Ixodidae), in St. Croix, U.S. Virgin Islands. Small mammals; birds; white-tailed deer, Odocoileus virginianus (Zimmermann); and feral cattle, Bos taurus L., were examined at nine premises, in mountainous rain forest, and in surrounding areas in western St. Croix, an area including and central to all known bont tick-infested premises on the island. Small Asian mongooses, Herpestes javanicus (E. Geoffroy Saint-Hilaire), yielded 1,566 ectoparasite specimens, representing five species, and including larvae of a soft tick, Carios puertoricensis (Fox); the tropical horse tick, Anocentor nitens (Neumann); and the southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini). Black rats, Rattus rattus L., yielded 144 specimens, representing six ectoparasite species, including C. puertoricensis. Of 25 bird species examined, seven yielded 116 ectoparasite specimens representing at least 14 different species of lice and mites, but no ticks. White-tailed deer and feral cattle yielded only various stages of A. nitens and R. microplus ticks. A. variegatum was not encountered on any potential wildlife host sampled, reflecting its low occurrence in St. Croix during the survey period. One collection of chewing lice (Phthiraptera: Philopteridae) from a spotted sandpiper, Actitis macularia (L.), and collections of feather mites (Acari: Astigmata: Trouessartiidae) from both bananaquits, Coereba flaveola (L.), and black-faced grassquits, Tiaris bicolor (L.), may represent new, undescribed species.

  3. Phylogeographic analysis reveals association of tick-borne pathogen, Anaplasma marginale, MSP1a sequences with ecological traits affecting tick vector performance

    PubMed Central

    Estrada-Peña, Agustín; Naranjo, Victoria; Acevedo-Whitehouse, Karina; Mangold, Atilio J; Kocan, Katherine M; de la Fuente, José

    2009-01-01

    Background The tick-borne pathogen Anaplasma marginale, which is endemic worldwide, is the type species of the genus Anaplasma (Rickettsiales: Anaplasmataceae). Rhipicephalus (Boophilus) microplus is the most important tick vector of A. marginale in tropical and subtropical regions of the world. Despite extensive characterization of the genetic diversity in A. marginale geographic strains using major surface protein sequences, little is known about the biogeography and evolution of A. marginale and other Anaplasma species. For A. marginale, MSP1a was shown to be involved in vector-pathogen and host-pathogen interactions and to have evolved under positive selection pressure. The MSP1a of A. marginale strains differs in molecular weight because of a variable number of tandem 23-31 amino acid repeats and has proven to be a stable marker of strain identity. While phylogenetic studies of MSP1a repeat sequences have shown evidence of A. marginale-tick co-evolution, these studies have not provided phylogeographic information on a global scale because of the high level of MSP1a genetic diversity among geographic strains. Results In this study we showed that the phylogeography of A. marginale MSP1a sequences is associated with world ecological regions (ecoregions) resulting in different evolutionary pressures and thence MSP1a sequences. The results demonstrated that the MSP1a first (R1) and last (RL) repeats and microsatellite sequences were associated with world ecoregion clusters with specific and different environmental envelopes. The evolution of R1 repeat sequences was found to be under positive selection. It is hypothesized that the driving environmental factors regulating tick populations could act on the selection of different A. marginale MSP1a sequence lineages, associated to each ecoregion. Conclusion The results reported herein provided the first evidence that the evolution of A. marginale was linked to ecological traits affecting tick vector performance. These

  4. Detection of bacteria related to Candidatus Midichloria mitochondrii in tick cell lines.

    PubMed

    Najm, Nour-Addeen; Silaghi, Cornelia; Bell-Sakyi, Lesley; Pfister, Kurt; Passos, Lygia M Friche

    2012-01-01

    Many ticks have been shown to be infected with intracellular bacteria. One of these bacteria is Candidatus Midichloria mitochondrii which is the only characterized bacterium that has the ability to invade the mitochondria within ovarian cells and consume them without any effect on the female tick's reproduction. In the present study, eight cell lines derived from the ticks Ixodes ricinus, Ixodes scapularis, Rhipicephalus (Boophilus) microplus, and Rhipicephalus (Boophilus) decoloratus were examined for the presence of the bacterium Ca. Midichloria mitochondrii. PCR assays for this bacterium were carried out using two sets of primers targeting the eubacterial 16SrRNA gene and a set of primers specific for the gyrB gene of Ca. Midichloria mitochondrii. With the 16S rRNA primers, DNA was amplified from two cell lines (R. (B.) decoloratus line BDE/CTVM14 and I. ricinus line IRE/CTVM19) on one out of three occasions each. Sequencing of the PCR products showed that the two cell lines gave sequences with 100% similarity to Ca. Midichloria mitochondrii. However, all cell lines, including the two positive cell lines, were negative with the specific primers. Phylogenetic analysis shows that our sequences belong to the subclass α-proteobacteria. They were identical to the sequences amplified from the tick I. ricinus. The results suggest that two cell lines, IRE/CTVM19 and BDE/CTVM14, may contain bacteria closely related to Ca. Midichloria mitochondrii and identical with it in a 350-bp part of the 16S rRNA gene sequence. To our knowledge, this constitutes the first report of the presence of DNA similar to the DNA of Ca. Midichloria mitochondrii in tick cell lines.

  5. Integrated Strategy for Sustainable Cattle Fever Tick Eradication in USA is Required to Mitigate the Impact of Global Change

    PubMed Central

    Pérez de León, Adalberto A.; Teel, Pete D.; Auclair, Allan N.; Messenger, Matthew T.; Guerrero, Felix D.; Schuster, Greta; Miller, Robert J.

    2012-01-01

    The ticks Rhipicephalus (Boophilus) annulatus and R. (B.) microplus, commonly known as cattle and southern cattle tick, respectively, impede the development and sustainability of livestock industries throughout tropical and other world regions. They affect animal productivity and wellbeing directly through their obligate blood-feeding habit and indirectly by serving as vectors of the infectious agents causing bovine babesiosis and anaplasmosis. The monumental scientific discovery of certain arthropod species as vectors of infectious agents is associated with the history of research on bovine babesiosis and R. annulatus. Together, R. microplus and R. annulatus are referred to as cattle fever ticks (CFT). Bovine babesiosis became a regulated foreign animal disease in the United States of America (U.S.) through efforts of the Cattle Fever Tick Eradication Program (CFTEP) established in 1906. The U.S. was declared free of CFT in 1943, with the exception of a permanent quarantine zone in south Texas along the border with Mexico. This achievement contributed greatly to the development and productivity of animal agriculture in the U.S. The permanent quarantine zone buffers CFT incursions from Mexico where both ticks and babesiosis are endemic. Until recently, the elimination of CFT outbreaks relied solely on the use of coumaphos, an organophosphate acaricide, in dipping vats or as a spray to treat livestock, or the vacation of pastures. However, ecological, societal, and economical changes are shifting the paradigm of systematically treating livestock to eradicate CFT. Keeping the U.S. CFT-free is a critical animal health issue affecting the economic stability of livestock and wildlife enterprises. Here, we describe vulnerabilities associated with global change forces challenging the CFTEP. The concept of integrated CFT eradication is discussed in reference to global change. PMID:22712018

  6. Ehrlichiae and ehrlichial diseases in china.

    PubMed

    Wen, Bohai; Cao, Wuchun; Pan, Hua

    2003-06-01

    The various ticks collected from different areas of China were examined for the existence of ehrlichial agents by polymerase chain reaction (PCR) with genus- or species-specific primers designed on the basis of ehrlichial 16S rRNA genes and sequence analyses. In southern China, E. chaffeensis was detected in Amblyomma testudinarium ticks from infested cattle, Haemaphysalis yeni ticks from hare, and Ixodes ovatus ticks from Muntiacus reevesi. E. canis was identified in Rhipicephalus sanguineus ticks from dogs and Boophilus microplus ticks from goats. A new species of the genus Ehrlichia, closely related to E. chaffeensis, and Anaplasma marginale were found in B. microplus ticks from cattle in Tibet. In northern China, E. chaffeensis was detected in Dermacentor silvarum and I. persulcatus ticks; the granulocytic ehrlichial agents were detected in I. persulcatus ticks from an area where Lyme disease is endemic. Canine ehrlichiosis was found in southern China and E. canis and E. platys were identified in dogs; human ehrlichioses were demonstrated by amplifying the 16S rRNA genes of E. chaffeensis and granulocytic ehrlichial agents from patients' blood specimens. In comparison of 16S rRNA gene sequences, the sequences of E. chaffeensis, E. canis, and E. platys in China were found to be different from that in other countries at certain nucleotide positions. These results reveal that a variety of tick-borne ehrlichial agents and diseases exist in China, and the ehrlichial agents and their tick-vectors are same as or different from that in other countries at species or strain levels.

  7. ATP Binding Cassette Transporter Mediates Both Heme and Pesticide Detoxification in Tick Midgut Cells

    PubMed Central

    Lara, Flavio Alves; Pohl, Paula C.; Gandara, Ana Caroline; Ferreira, Jessica da Silva; Nascimento-Silva, Maria Clara; Bechara, Gervásio Henrique; Sorgine, Marcos H. F.; Almeida, Igor C.; Vaz, Itabajara da Silva; Oliveira, Pedro L.

    2015-01-01

    In ticks, the digestion of blood occurs intracellularly and proteolytic digestion of hemoglobin takes place in a dedicated type of lysosome, the digest vesicle, followed by transfer of the heme moiety of hemoglobin to a specialized organelle that accumulates large heme aggregates, called hemosomes. In the present work, we studied the uptake of fluorescent metalloporphyrins, used as heme analogs, and amitraz, one of the most regularly used acaricides to control cattle tick infestations, by Rhipicephalus (Boophilus) microplus midgut cells. Both compounds were taken up by midgut cells in vitro and accumulated inside the hemosomes. Transport of both molecules was sensitive to cyclosporine A (CsA), a well-known inhibitor of ATP binding cassette (ABC) transporters. Rhodamine 123, a fluorescent probe that is also a recognized ABC substrate, was similarly directed to the hemosome in a CsA-sensitive manner. Using an antibody against conserved domain of PgP-1-type ABC transporter, we were able to immunolocalize PgP-1 in the digest vesicle membranes. Comparison between two R. microplus strains that were resistant and susceptible to amitraz revealed that the resistant strain detoxified both amitraz and Sn-Pp IX more efficiently than the susceptible strain, a process that was also sensitive to CsA. A transcript containing an ABC transporter signature exhibited 2.5-fold increased expression in the amitraz-resistant strain when compared with the susceptible strain. RNAi-induced down-regulation of this ABC transporter led to the accumulation of metalloporphyrin in the digestive vacuole, interrupting heme traffic to the hemosome. This evidence further confirms that this transcript codes for a heme transporter. This is the first report of heme transport in a blood-feeding organism. While the primary physiological function of the hemosome is to detoxify heme and attenuate its toxicity, we suggest that the use of this acaricide detoxification pathway by ticks may represent a new

  8. ATP Binding Cassette Transporter Mediates Both Heme and Pesticide Detoxification in Tick Midgut Cells.

    PubMed

    Lara, Flavio Alves; Pohl, Paula C; Gandara, Ana Caroline; Ferreira, Jessica da Silva; Nascimento-Silva, Maria Clara; Bechara, Gervásio Henrique; Sorgine, Marcos H F; Almeida, Igor C; Vaz, Itabajara da Silva; Oliveira, Pedro L

    2015-01-01

    In ticks, the digestion of blood occurs intracellularly and proteolytic digestion of hemoglobin takes place in a dedicated type of lysosome, the digest vesicle, followed by transfer of the heme moiety of hemoglobin to a specialized organelle that accumulates large heme aggregates, called hemosomes. In the present work, we studied the uptake of fluorescent metalloporphyrins, used as heme analogs, and amitraz, one of the most regularly used acaricides to control cattle tick infestations, by Rhipicephalus (Boophilus) microplus midgut cells. Both compounds were taken up by midgut cells in vitro and accumulated inside the hemosomes. Transport of both molecules was sensitive to cyclosporine A (CsA), a well-known inhibitor of ATP binding cassette (ABC) transporters. Rhodamine 123, a fluorescent probe that is also a recognized ABC substrate, was similarly directed to the hemosome in a CsA-sensitive manner. Using an antibody against conserved domain of PgP-1-type ABC transporter, we were able to immunolocalize PgP-1 in the digest vesicle membranes. Comparison between two R. microplus strains that were resistant and susceptible to amitraz revealed that the resistant strain detoxified both amitraz and Sn-Pp IX more efficiently than the susceptible strain, a process that was also sensitive to CsA. A transcript containing an ABC transporter signature exhibited 2.5-fold increased expression in the amitraz-resistant strain when compared with the susceptible strain. RNAi-induced down-regulation of this ABC transporter led to the accumulation of metalloporphyrin in the digestive vacuole, interrupting heme traffic to the hemosome. This evidence further confirms that this transcript codes for a heme transporter. This is the first report of heme transport in a blood-feeding organism. While the primary physiological function of the hemosome is to detoxify heme and attenuate its toxicity, we suggest that the use of this acaricide detoxification pathway by ticks may represent a new

  9. Infection and seroconversion of susceptible animals introduced into a babesiosis endemic area.

    PubMed

    Lopez, Martin; Figueroa, Julio V; Ramos, Juan A; Mosqueda, Juan J; Rojas, Edmundo; Vega, Carlos A; Alvarez, J Antonio

    2008-12-01

    The study aimed to determine the incubation period of Babesia sp. infection in naive cattle and to monitor the serological response once exposed to natural Boophilus microplus (Rhipicephalus microplus)-infested paddocks. The study was carried out on a farm located in Veracruz, Mexico. Five groups of five steers were relocated every 3 months from a tick-free area to a tick-infested paddock. Animals were introduced in October, January, April, July, and October. Blood samples were taken daily until day 21 to determine packed cell volume (PCV), percentage of parasitized erythrocytes (PPE), and antibody titers to Babesia bigemina and B. bovis by the indirect fluorescent antibody procedure. Detection of Babesia in blood was also performed by species-specific PCR. The estimated incubation period was 6-14 days post introduction to paddocks (PIP), with fever (41 degrees C) for at least 3 days. PCV decreased by >25% and Babesia parasites were observed during the clinical phase of the disease. The highest individual PPEs (0.44% and 0.22% for B. bovis and B. bigemina, respectively) were observed from animals introduced in October. The four other groups showed a mean PPE ranging from 0.002-0.146% at day 14 PIP. All animals were detected as PCR positive between 8-14 days PIP. The highest antibody titers were 1:3328. The environmental conditions were favorable throughout the year for tick reproduction as the farm showed enzootic stability and hyperendemic conditions for bovine babesiosis. In this type of farm, strategic tick control could be accompanied by babesiosis vaccination, particularly for cattle relocated from tick-free areas.

  10. Temporal characterization of the organ-specific Rhipicephalus microplus transcriptional response to Anaplasma marginale infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Arthropods transmit a variety of important infectious diseases of humans and animals. Importantly, replication and development of pathogen infectivity is tightly linked to vector feeding on the mammalian host; thus analysis of the transcriptomes of both vector and pathogen during feeding is fundamen...

  11. Expression of 6-Cys gene superfamily defines babesia bovis sexual stage development within rhipicephalus microplus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Babesia bovis, an intra-erythrocytic tick-borne apicomplexan protozoan, is one of the agents of bovine babesiosis. Its life cycle includes sexual reproduction within cattle fever ticks, Rhipicephalus spp. Six B. bovis 6-Cys gene superfamily members were previously identified (A, B, C, D, E, F) and t...

  12. Analysis of stage specific protein expression during babesia bovis development within rhipicephalus microplus female ticks

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Arthropod borne pathogens have a complex life cycle that includes asexual reproduction of haploid stages in mammalian erythrocytes and development of diploid stages in the vector. Transition of Apicomplexan pathogens between the mammalian host and the arthropod vector is critical for ongoing transmi...

  13. Acaricidal, pediculocidal and larvicidal activity of synthesized ZnO nanoparticles using wet chemical route against blood feeding parasites.

    PubMed

    Kirthi, Arivarasan Vishnu; Rahuman, Abdul Abdul; Rajakumar, Govindasamy; Marimuthu, Sampath; Santhoshkumar, Thirunavukkarasu; Jayaseelan, Chidambaram; Velayutham, Kanayairam

    2011-08-01

    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

  14. Co-transmission of the non-transmissible South African Babesia bovis S24 vaccine strain during mixed infection with a field isolate.

    PubMed

    Combrink, M P; Troskie, P C; de Klerk, D G; Pienaar, R; Latif, A A; Mans, B J

    2015-03-01

    The South African Babesia bovis live blood vaccine, originating from a field isolate attenuated by 23 serial syringe passages in splenectomized calves, has lost the ability to infect the natural vector Rhipicephalus (Boophilus) microplus. In this study, infection with mixed parasites from the vaccine strain and a field isolate, resulted in transmission of both genotype populations. Comparing the field isolate and transmitted combination indicated no significant difference in their virulence, while challenge of vaccinated cattle with these isolates showed the ability of the vaccine to protect against both. Limiting dilution of the transmitted combination, followed by infection of splenectomized cattle (n=34) yielded no single infections for the vaccine strain genotype, seven clonal lines of the field isolate and one mixture of vaccine strain and field isolate. Only one of two field isolate clonal lines selected for vector transmission study was transmitted. Showing that B. bovis isolates can contain both tick transmissible and non-transmissible subpopulations. The findings of this study also indicate the probability of vaccine co-infection transmission occurring in the field, which may result in new genotype populations of B. bovis. However, the impact of this recombination with field isolates is considered negligible since a genotypically diverse population of B. bovis is already present in South Africa.

  15. Hemalin, a thrombin inhibitor isolated from a midgut cDNA library from the hard tick Haemaphysalis longicornis.

    PubMed

    Liao, Min; Zhou, Jinlin; Gong, Haiyan; Boldbaatar, Damdinsuren; Shirafuji, Rika; Battur, Banzragch; Nishikawa, Yoshifumi; Fujisaki, Kozo

    2009-02-01

    A full-length sequence of a thrombin inhibitor (designated as hemalin) from the midgut of parthenogenetic Haemaphysalis longicornis has been identified. Sequence analysis shows that this gene belongs to the Kunitz-type family, containing two Kunitz domains with high homology to boophilin, the thrombin inhibitor from Rhipicephalus (Boophilus) microplus. The recombinant protein expressed in insect cells delayed bovine plasma clotting time and inhibited both thrombin-induced fibrinogen clotting and platelet aggregation. A 20-kDa protein was detected from the midgut lysate with antiserum against recombinant hemalin. The gene is expressed at all stages of the tick except for the egg stage, and hemalin mRNA mainly in the midgut of the female adult tick. Real-time PCR analysis shows that this gene has a distinctly high expression level in the rapid bloodsucking period of the larvae, nymphs, and adults. Disruption of the hemalin gene by RNA interference led to a 2-day extension of the tick blood feeding period, and 27.7% of the RNA-treated ticks did not successfully complete the blood feeding. These findings indicate that the newly identified thrombin inhibitor from the midgut of H. longicornis might play an important role in tick blood feeding.

  16. On the search for markers of tick resistance in bovines.

    PubMed

    Regitano, L C A; Ibelli, A M G; Gasparin, G; Miyata, M; Azevedo, A L S; Coutinho, L L; Teodoro, R L; Machado, M A; Silva, M V G B; Nakata, L C; Zaros, L G; Sonstegard, T S; Silva, A M; Alencar, M M; Oliveira, M C S

    2008-01-01

    Genetic differences in susceptibility to ticks (Rhipicephalus (Boophilus) microplus) are considerable in bovines. Here, mapping, association and gene expression approaches were employed to further advance our understanding of the molecular basis of tick resistance. A B. taurus x B. indicus F2 population was developed by Embrapa and 382 individuals were measured for parasitic load. Scanning of all chromosomes is in progress. Quantitative trait loci (QTL) for tick load were mapped to chromosomes 4, 5, 7, 10, 14, 18 and 23 out of the 20 chromosomes scanned and were dependent on the season in which the phenotype was scored. In the candidate gene approach, females from the genetic groups Nelore (NE--184), Canchim x Nelore (CN--153), Aberdeen Angus x Nelore (AN--123) and Simmental x Nelore (SN--120) were evaluated under natural infestation. Microsatellite markers close to the genes for interleukin 2 (IL2), interleukin 4 (IL4) and interferon gamma (IFNG) were analysed. Tick counts were associated with the marker for interleukin 4 (P < 0.05) in three genetic groups. Differences in cytokine mRNA levels of naive versus infested Nelore calves as well as between resistant versus susceptible cows from NE, CN and AN genetic groups were also investigated. Comparison of cytokines from infested and naïve animals showed downregulation of IL2. When resistant cows were compared to susceptible animals, IL8 was downregulated. These results reinforce the multiloci nature of tick resistance and the need to consider QTL and environment interactions.

  17. Perceptions and attitudes among milk producers in Minas Gerais regarding cattle tick biology and control.

    PubMed

    do Amaral, Maria Alice Zacarias; da Rocha, Christiane Maria Barcellos Magalhães; Faccini, João Luiz; Furlong, John; Monteiro, Caio Márcio de Oliveira; Prata, Márcia Cristina de Azevedo

    2011-01-01

    This study evaluates milk producers' knowledge regarding cattle ticks and practices for controlling them. Ninety-three dairymen in Minas Gerais were interviewed. These producers had no information regarding acaricide efficiency tests. To analyze the information, open responses were categorized through "content analysis", and descriptive analysis consisting of extracting the profile highlighted by the highest frequencies. The association between schooling level and knowledge was tested by means of chi-square trend tests. It was observed that 92.3% had no knowledge of the nonparasitic period. For 96.4%, what determined the time to apply treatment was the degree of tick infestation; 93.3% used spray guns to apply the acaricide. In seeking to cross-correlate the biological and control variables with education, cooperative action, length of experience and herd size, it was found that there was a linear association between schooling level and implementation of acaricide solution preparation. The other factors didn't show any significant association. These data demonstrated the need to instruct the producers in relation to the biology and control of Rhipicephalus (Boophilus) microplus. It was concluded that the majority of milk producers were unaware of cattle tick biology and the factors that influence choosing an acaricide, which makes it difficult to implement strategic control.

  18. Seasonal Variation and Frequency Distribution of Ectoparasites in Crossbreed Cattle in Southeastern Brazil

    PubMed Central

    Ferraz da Costa, Maria do Socorro; Guimarães, Marcos Pezzi; Lima, Walter dos Santos; Ferraz da Costa, Ana Julia; Facury Filho, Elias Jorge; Araujo, Ricardo Nascimento

    2014-01-01

    The aims of this study were to evaluate the seasonal variation and frequency distribution of Rhipicephalus (Boophilus) microplus, Haematobia irritans, and Dermatobia hominis on crossbred heifers under field conditions in the northeast of Minas Gerais state, southeastern Brazil. From November 2007 to September 2009 (23 months), 40 heifers aged 16.6 ± 2.4 months were divided into groups A (1/4 Holstein × 3/4 Gir) and B (1/2 Holstein × 1/2 Gir) and had the monthly infestation estimated along with the climatic conditions. The mean maximum and minimum temperatures were 28.5 and 19°C, respectively. The ectoparasites were present on animals in all months of the year. The levels of ticks on the animals were low (3.0 ± 0.2 ticks/animal), with the highest density in midwinter. The temperature was the climatic factor that most influenced the tick levels. The population of H. irritans (13.9 ± 0.3 flies/animal) and D. hominis (1.5 ± 0.2 larvae/animal) on heifers was more influenced by rainfall and exhibited two population peaks during the year. 1/2 Holstein heifers harbored significantly more H. irritans and D. hominis than 1/4 Holstein heifers. The results are discussed considering the most appropriate periods to apply ectoparasiticides and the genetic make-up of the animals. PMID:26464941

  19. [Compatibility of Beauveria bassiana (Ascomycetes: Clavicipitaceae) with chemicals acaricides used in the control of cattle tick].

    PubMed

    Barci, Leila A G; Wenzel, Inajá M; de Almeida, José Eduardo M; de Campos Nogueira, Adriana H; do Prado, Angelo P

    2009-12-01

    The purpose of the present study was to assess compatibility between IBCB66 and IBCB21 isolates of Beauveria bassiana and acaricides: Flumethrin+Coumaphos, Deltamethrin, Dichlorvos+Cypermethrin, Dichlorvos+Chlorpyrifos, Cypermethrin High Cis, Dichlorvos+Cypermethrin High Cis, Cypermethrin and Amitraz, utilized on the control of Rhipicephalus (Boophilus) microplus in our country. The effect of commercial products on the isolates was assayed according to observation of vegetative growth, conidia production, and viability of strains of B. bassiana fungus. With concerning about IBCB66 isolate, products Deltamethrin, Cypermethrin High Cis and Amitraz were compatible, not affecting the entomopathogen development. Product Cypermethrin was toxic, and products Flumethrin+Coumaphos, Dichlorvos+Cypermethrin, Dichlorvos+Chlorpyrifos and Dichlorvos+Cypermethrin High Cis were very toxic. In regard to IBCB21 isolate, products Flumethrin+Coumaphos, Dichlorvos+Cypermethrin, Dichlorvos+Chlorpyrifos, Cypermethrin High Cis, Dichlorvos+Cypermethrin High Cis and Cypermethrin were very toxic and product Amitraz was toxic. From the acaricides evaluated, product Deltamethrin was the single agent that did not produce toxic effect on the entomopathogen.

  20. Effects of microplusin, a copper-chelating antimicrobial peptide, against Cryptococcus neoformans.

    PubMed

    Silva, Fernanda D; Rossi, Diego C P; Martinez, Luis R; Frases, Susana; Fonseca, Fernanda L; Campos, Claudia Barbosa L; Rodrigues, Marcio L; Nosanchuk, Joshua D; Daffre, Sirlei

    2011-11-01

    Microplusin is an antimicrobial peptide isolated from the cattle tick Rhipicephalus (Boophilus) microplus. Its copper-chelating ability is putatively responsible for its bacteriostatic activity against Micrococcus luteus as microplusin inhibits respiration in this species, which is a copper-dependent process. Microplusin is also active against Cryptococcus neoformans (MIC(50) = 0.09 μM), the etiologic agent of cryptococcosis. Here, we show that microplusin is fungistatic to C. neoformans and this inhibitory effect is abrogated by copper supplementation. Notably, microplusin drastically altered the respiratory profile of C. neoformans. In addition, microplusin affects important virulence factors of this fungus. We observed that microplusin completely inhibited fungal melanization, and this effect correlates with the inhibition of the related enzyme laccase. Also, microplusin significantly inhibited the capsule size of C. neoformans. Our studies reveal, for the first time, a copper-chelating antimicrobial peptide that inhibits respiration and growth of C. neoformans and modifies two major virulence factors: melanization and formation of a polysaccharide capsule. These features suggest that microplusin, or other copper-chelation approaches, may be a promising therapeutic for cryptococcosis.

  1. Repellent effects of Melaleuca alternifolia (tea tree) oil against cattle tick larvae (Rhipicephalus australis) when formulated as emulsions and in β-cyclodextrin inclusion complexes.

    PubMed

    Yim, Wei Tsun; Bhandari, Bhesh; Jackson, Louise; James, Peter

    2016-07-30

    Rhipicephalus australis (formerly Boophilus microplus) is a one host tick responsible for major economic loss in tropical and subtropical cattle production enterprises. Control is largely dependent on the application of acaricides but resistance has developed to most currently registered chemical groups. Repellent compounds that prevent initial attachment of tick larvae offer a potential alternative to control with chemical toxicants. The repellent effects of Melaleuca alternifolia oil (TTO) emulsions and two β-cyclodextrin complex formulations, a slow release form (SR) and a modified faster release form (FR), were examined in a series of laboratory studies. Emulsions containing 4% and 5% TTO applied to cattle hair in laboratory studies completely repelled ascending tick larvae for 24h whereas 2% and 3% formulations provided 80% protection. At 48h, 5% TTO provided 78% repellency but lower concentrations repelled less than 60% of larvae. In a study conducted over 15 days, 3% TTO emulsion applied to cattle hair provided close to 100% repellency for 2 days, but then protection fell to 23% by day 15. The FR formulation gave significantly greater repellency than the emulsion and the SR formulation from day 3 until the end of the study (P<0.05), providing almost complete repellency at day 3 (99.5%), then decreasing over the period of the study to 49% repellency at day 15. Proof of concept is established for the use of appropriately designed controlled-release formulations to extend the period of repellency provided by TTO against R. australis larvae.

  2. Immunisation with recombinant proteins subolesin and Bm86 for the control of Dermanyssus gallinae in poultry.

    PubMed

    Harrington, David; Canales, Mario; de la Fuente, José; de Luna, Carlos; Robinson, Karen; Guy, Jonathan; Sparagano, Olivier

    2009-06-19

    Dermanyssus gallinae has a worldwide distribution and is considered to be the most serious and economically significant ectoparasite affecting egg-laying poultry in Europe. Recombinant Bm86 and subolesin proteins derived from Boophilus microplus ticks and Aedes albopictus mosquitoes were used to immunise poultry in an attempt to control D. gallinaein vitro. Immunisation with subolesin and Bm86 stimulated different profiles of IgY response, whilst Bm86 but not subolesin was recognized by IgY on western blots. Orthologues for Bm86 were not found in D. gallinae by PCR, but a 150 bp fragment aligned with mammalian akirin 1 and a 300 bp fragment aligned with Amblyomma hebraeum were amplified by subolesin PCR. D. gallinae mortality after feeding was 35.1% higher (P=0.009) in the Subolesin group and 23% higher (not significant) in the Bm86 compared to the Control group. Thus it can be concluded that immunisation with recombinant subolesin can stimulate a protective response in laying hens against D. gallinae.

  3. A house dust mite allergen homologue from poultry red mite Dermanyssus gallinae (De Geer).

    PubMed

    Nisbet, A J; Huntley, J F; Mackellar, A; Sparks, N; McDevitt, R

    2006-08-01

    Tropomyosin is an allergenic, actin-binding protein and a proposed vaccine candidate from several species of parasite. Tropomyosin cDNA, obtained by polymerase chain reaction (PCR) amplification from Dermanyssus gallinae RNA, encoded a predicted protein with 89% and 88% identity to tropomyosins from the ticks Boophilus microplus and Haemaphysalis longicornis, respectively, and 85% identity to the house dust mite (HDM) tropomyosin Der p 10. Mouse antibodies raised against HDM tropomyosin reacted with a band of 38 kDa on Western blots of D. gallinae extract, consistent with the molecular masses of acarine tropomyosins and the putative product of the cDNA encoding D. gallinae tropomyosin. When the same preparation of D. gallinae proteins was used in Western blots with serum from infested hens, the IgY component of the serum bound to a number of mite proteins, but not to tropomyosin, indicating that hens are not directly exposed to this allergen during a natural infestation. Immunolocalization of tropomyosin in mites indicated a ubiquitous distribution of the molecule in mite tissues. Immunolocalization and Western blotting also indicated that poultry red mites ingest host IgY.

  4. Ectoparasites of dogs in home environments on the Caribbean slope of Costa Rica.

    PubMed

    Troyo, Adriana; Calderón-Arguedas, Olger; Alvarado, Gilbert; Vargas-Castro, Luis E; Avendaño, Adrián

    2012-01-01

    Reports on ectoparasites on dogs in Central America are scarce. The aim of this study was to identify flea, louse and tick species infesting dogs in home environments on the Caribbean slope of Costa Rica, and determine their frequency and coexistence. Ectoparasites were collected from dogs in 83 rural homes at five study sites. Specimens were identified and separated according to species. Fleas were the most common ectoparasite (G = 22,217, DF = 8, p = 0.004). Ctenocephalides felis and Pulex simulans were found in 83% and 55% of the homes with ectoparasites, respectively. Trichodectes canis (13%), Heterodoxus spiniger (10%), Rhipicephalus sanguineus (18%), Rhipicephalus (Boophilus) microplus (5%) and Amblyomma ovale (8%) were also present. More than one species was collected in most cases (66%), and the most common combination was C. felis and P. simulans (59% of homes with fleas). The high frequency of P. simulans emphasizes the need for adequate identification. This was the first study involving different ectoparasites of dogs in Costa Rica, as well as the first report of T. canis in this country. The relative frequency and coexistence of these ectoparasites in the home environment may have implications for animal and human health.

  5. Immunological control of ectoparasites: past achievements and future research priorities.

    PubMed

    Willadsen, P

    1999-11-01

    Recombinant vaccines are available for the control of the tick Boophilus microplus, while progress has been made in the development of vaccines against Lucilia cuprina and Chrysomya bezziana. Literature suggests that the control of other ectoparasites is feasible, either through the duplication in a vaccine of naturally acquired immunity or through 'concealed' antigen vaccines. Major deficiencies in our current knowledge however point to possible research opportunities for the future. The identification of protective antigens from all species is proceeding slowly, particularly for the antigens of naturally acquired immunity. Our capacity to produce effective recombinant antigens has progressed greatly, though there remains a major difficulty where some or all of the protective effect is due to immunogenic oligosaccharide. Our understanding of protective mechanisms is limited. The delivery of the appropriate immunological response remains difficult. Nevertheless, some of the most critical areas of ignorance are in basic biological issues: factors which affect the susceptibility of particular pest species to immunological attack and the implications of vaccine-induced effects for pest and disease control under field conditions. Increasingly too, effective pest control is likely to demand the integration of a variety of control technologies. The study of this integration is in its infancy.

  6. Vaccination against ectoparasites.

    PubMed

    Willadsen, P

    2006-01-01

    Ectoparasites of livestock are of great economic and social importance but their effective control remains difficult. The feasibility of vaccination as a novel control measure was established over a decade ago with the commercial release of a recombinant vaccine against the cattle tick Boophilus microplus. Since then, research has continued on ticks and other ectoparasites. While some ectoparasite species will undoubtedly be refractory to immunological control, for others there has been a steady accumulation of knowledge of partially protective antigens, now accelerating through the application of genomic technologies. Nevertheless, progress towards usable, commercially available vaccines has been limited by a number of factors. The number of highly effective antigens is still very small. Although some classes of antigen have been investigated in more detail than others, we have no systematic knowledge of what distinguishes an effective antigen. Much hope has been placed on the potential of multi-antigen mixtures to deliver the efficacy required of a successful vaccine but with little experimental evidence. The application of current knowledge across parasite and host species needs to be explored but little has been done. In most cases, the path to commercial delivery is uncertain. Although many constraints and challenges remain, the need for vaccines and our capacity to develop them can only increase.

  7. Detection of Rickettsia parkeri from within Piura, Peru, and the First Reported Presence of Candidatus Rickettsia andeanae in the Tick Rhipicephalus sanguineus

    PubMed Central

    Flores-Mendoza, Carmen; Florin, David; Felices, Vidal; Pozo, Edwar J.; Graf, Paul C.F.; Richards, Allen L.

    2013-01-01

    Abstract Domestic farm animals (n=145) were sampled for the presence of ectoparasites in northwestern Peru during March, 2008. Ninety domestic animals (62%) were positive for the presence of an ectoparasite(s) and produced a total collection of the following: 728 ticks [Amblyomma maculatum, Anocentor nitens, Rhipicephalus (Boophilus) microplus, Rhipicephalus sanguineus, and Otobius megnini], 12 lice (Haematopinus suis), and 3 fleas (Ctenocephalides felis). A Rickettsia genus-specific qPCR assay was performed on nucleic acid preparations of the collected ectoparasites that resulted in 5% (37/743, 35 ticks and 2 fleas) of the ectoparasites positive for the presence of Rickettsia. DNA from the positive individual ticks was tested with 2 other qPCR assays for the presence of the ompB gene in Candidatus Rickettsia andeanae or Rickettsia parkeri. Candidatus R. andeanae was found in 25 A. maculatum ticks and in two Rh. sanguineus ticks, whereas R. parkeri was detected in 6 A. maculatum ticks. Two A. maculatum were co-infected with both Candidatus R. andeanae and R. parkeri. Rickettsia felis was detected in 2 fleas, Ctenocephalides felis, by multilocus sequence typing of the 17-kD antigen and ompA genes. These findings expand the geographic range of R. parkeri to include Peru as well as expand the natural arthropod vector of Candidatus R. andeanae to include Rhipicephalus sanguineus. PMID:23488453

  8. Experimental vaccination of sheep and cattle against tick infestation using recombinant 5'-nucleotidase.

    PubMed

    Hope, M; Jiang, X; Gough, J; Cadogan, L; Josh, P; Jonsson, N; Willadsen, P

    2010-02-01

    Limited prior evidence suggests that 5'-nucleotidase, an ectoenzyme principally located in the Malpighian tubules of the tick Rhipicephalus (Boophilus) microplus, could be an effective antigen in an anti-tick vaccine. To assess this, recombinant 5'-nucleotidase was expressed in Escherichia coli and used in vaccination trials with both sheep and cattle. Vaccinated sheep were challenged with freshly moulted adult ticks. Those with high titres of anti-nucleotidase antibodies showed significant protection against tick infestation, although protection was less than that found with the previously characterized antigen, Bm86. Cattle were vaccinated, in separate groups, with 5'-nucleotidase, Bm86 and both antigens combined. Cattle, as the natural host, were challenged with larval ticks. Although Bm86 showed typical efficacy, no significant protection was seen in cattle vaccinated with 5'-nucleotidase. Cattle receiving a dual antigen formulation were no better protected than those receiving Bm86 alone. One possible reason for the difference between host species, namely antibody titre, was examined and shown to be an unlikely explanation. This demonstrates a limitation of using a model host like sheep in vaccine studies.

  9. Experimental vaccination of sheep and cattle against tick infestation using recombinant 5′-nucleotidase

    PubMed Central

    HOPE, M; JIANG, X; GOUGH, J; CADOGAN, L; JOSH, P; JONSSON, N; WILLADSEN, P

    2010-01-01

    Limited prior evidence suggests that 5′-nucleotidase, an ectoenzyme principally located in the Malpighian tubules of the tick Rhipicephalus (Boophilus) microplus, could be an effective antigen in an anti-tick vaccine. To assess this, recombinant 5′-nucleotidase was expressed in Escherichia coli and used in vaccination trials with both sheep and cattle. Vaccinated sheep were challenged with freshly moulted adult ticks. Those with high titres of anti-nucleotidase antibodies showed significant protection against tick infestation, although protection was less than that found with the previously characterized antigen, Bm86. Cattle were vaccinated, in separate groups, with 5′-nucleotidase, Bm86 and both antigens combined. Cattle, as the natural host, were challenged with larval ticks. Although Bm86 showed typical efficacy, no significant protection was seen in cattle vaccinated with 5′-nucleotidase. Cattle receiving a dual antigen formulation were no better protected than those receiving Bm86 alone. One possible reason for the difference between host species, namely antibody titre, was examined and shown to be an unlikely explanation. This demonstrates a limitation of using a model host like sheep in vaccine studies. PMID:20070827

  10. Acaricidal properties of the formulations based on essential oils from Cymbopogon winterianus and Syzygium aromaticum plants.

    PubMed

    de Mello, Valéria; Prata, Márcia Cristina de Azevedo; da Silva, Márcio Roberto; Daemon, Erik; da Silva, Luciane Santos; Guimarães, Flávia del Gaudio; de Mendonça, Alessandra Esther; Folly, Evelize; Vilela, Fernanda Maria Pinto; do Amaral, Lilian Henriques; Cabral, Lucio Mendes; do Amaral, Maria da Penha Henriques

    2014-12-01

    The cattle tick, Rhipicephalus (Boophilus) microplus, has caused serious harm to livestock raising in Brazil, considering the costs of controlling it, loss of revenue due to smaller production of milk and meat, and damage to leather, in addition to transmitting diseases. The use of medicinal plants is considered an alternative to the recurring resistance to chemicals. Due to the need for efficient alternatives with less environmental impact, this study aimed to develop contact formulations with essential oils from the Java citronella (Cymbopogon winterianus) and clove (Syzygium aromaticum) plants and to assess in vitro the effects in different stages of the tick cycle. In the present study, concentrations from 0.5-15.0% of the essential oils incorporated in the formulations were used. The ticks from different geographical areas were treated with those formulations, and their effects on the production levels of eggs, on the larvae hatching, and their efficiency on ticks were assessed. The obtained results were compared with other commercial acaricidal products. After the 20th day of treatment, the formulations with citronella essential oil had 2.09-55.51% efficiency, depending on the concentration of the oil incorporated. The efficiency of the treatment with formulations containing clove essential oil was higher, from 92.47-100%. The results showed the acaricidal effects of the formulations tested when compared to commercial chemical products. In vivo studies should be performed in order to assess the efficiency of those formulations in the fields, aiming to use these products as an alternative for controlling cattle ticks.

  11. [Ectoparasites in goat and sheep folks from Alto Mearim and Grajaú Microregion, State of Maranhão].

    PubMed

    Brito, Danilo R B; Santos, Ana Clara G; Guerra, Rita M S N C

    2005-01-01

    Goat and sheep folks from Alto Mearim and Grajaú microregion, State of Maranhão, were examined by inspection and by feeling to collected ectoparasites. The specimens were kept in individual bottles for each host sampled with alcohol 70 degrees GL as preserver liquid. The identification was carried out at Universidade Estadual do Maranhão at the laboratory of parasitology. The ticks were examined in stereomicroscope and identified through Aragão and Fonseca (1961) dicotomic key; the lice were mounted in slides according to Pinto (1938) technique, examined in optic microscope and identified through Tuff (1977) and Furmam and Catts (1977) keys; the fly larvae were examined in stereomicroscope and identified by the morphology of the peritreme, and spiracular slits. In 380 animals sampled, 143 (37,64%) were infested with ectoparasites. The following parasites were identified: Bovicola caprae, Boophilus microplus, larvae of Cochliomya hominivorax (screw-worm) and larvae of Dermatobia hominis (bot fly). Parasitary association was observed only in goats in Grajaú and Sítio Novo (louse+tick; louse+bot fly; screw-worm+louse and ticks+louse+screw-worm). The statistic analyses demonstrated that the prevalence of ectoparasites was high in the rainy season (P<0.02).

  12. Management and control of parasites on dairy farms in northwestern region of São Paulo state.

    PubMed

    Veríssimo, Cecília José; Vasques, Flávia; Duarte, Keila Maria Roncato; Paulino, Valdinei Tadeu; Ambrósio, Luis Alberto

    2016-01-01

    Dairy cattle farming is of great economic and social importance in all Brazilian's regions. Parasites can reduce milk productivity, especially the tick Rhipicephalus (Boophilus) microplus. This study consisted of a questionnaire answered by 40 milk producers in the northwestern region of the State of São Paulo. The aim was to ascertain how these producers controlled ticks and other parasites. Very many of them knew nothing about the biological cycle of the cattle tick or about strategic control or acaricide efficacy tests. The majority (87.5%) controlled ticks at a high frequency, without technical criteria and care to apply the acaricide. Spraying was the most used mode of acaricide application (95%) and endectocides were used by 45%. Cattle tick fever was the harm most associated with ticks (87.5%) followed closely by screwworm (77.5%). However, 65% were satisfied with their tick control. About the control of others parasites, all dewormed at least twice a year their animals; 65% were controlling horn fly; 40% had problems with screwworm. The interviewers had in general good level of education and the farms generally exhibited a high degree of technology for milk production on pasture because half of them received technical assistance frequently.

  13. Acaricidal, insecticidal, and larvicidal efficacy of fruit peel aqueous extract of Annona squamosa and its compounds against blood-feeding parasites.

    PubMed

    Madhumitha, Gunabalan; Rajakumar, Govindasamy; Roopan, Selvaraj Mohana; Rahuman, Abdul Abdul; Priya, Kanagaraj Mohana; Saral, Antoneyraj Mary; Khan, Fazlur Rahman Nawaz; Khanna, Venkatesh Gopiesh; Velayutham, Kannaiyaram; Jayaseelan, Chidambaram; Kamaraj, Chinnaperumal; Elango, Gandhi

    2012-11-01

    Plant products may be alternative sources of parasitic control agents, since they constitute a rich source of bioactive compounds that are eco-friendly and nontoxic products. The plant extracts are good and safe alternatives due to their low toxicity to mammals and easy biodegradability. In the present study, fruit peel aqueous extract of Annona squamosa (Annonaceae) extracted by immersion method exhibited adulticidal activity against Haemaphysalis bispinosa (Acarina: Ixodidae) and the hematophagous fly, Hippobosca maculata (Diptera: Hippoboscidae), and larvicidal activity against the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae), Anopheles subpictus, and Culex quinquefasciatus (Diptera: Culicidae). The chemical composition of A. squamosa fruit peel aqueous extract was analyzed by gas chromatography-mass spectrometry. The major chemical constituent of peel aqueous extract of A. squamosa was identified as 1H- cycloprop[e]azulen-7-ol decahydro-1,1,7-trimethyl-4-methylene-[1ar-(1aα,4aα, 7β, 7 a, β, 7bα)] (28.55%) by comparison of mass spectral data and retention times. The other major constituents present in the aqueous extract were retinal 9-cis- (12.61%), 3,17-dioxo-4-androsten-11alpha-yl hydrogen succinate (6.86%), 1-naphthalenepentanol decahydro-5-(hydroxymethyl)-5,8a-dimethyl-y,2-bis(methylene)-(1α,4aβ,5α,8aα) (14.83%), 1-naphthalenemethanol decahydro -5-(5-hydroxy-3-methyl-3-pentenyl)- 1,4a-di methyl - 6-methylene -(1S-[1α, 4aα, 5α(E), 8aβ] (4.44%), (-)-spathulenol (20.75%), podocarp-7-en-3-one13β-methyl-13-vinyl- (5.98%), and 1-phenanthrene carboxaldehyde 7-ethenyl-1,2,3,4,4a,4,5,6,7,9,10,10a-dodecahydro-1,4a,7-trimethyl-[1R-(1α,4aβ.4bα,7β, 10aα)]-(5.98%). The adult and larval parasitic mortalities observed in fruit peel aqueous extract of A. squamosa were 31, 59, 80, 91, and100%; 27, 42, 66, 87, and 100%; and 33, 45, 68, 92, and 100% at the concentrations of 250, 500, 1,000, 1,500, and 2,000 ppm, respectively, against

  14. Assessment of Acute Oral and Dermal Toxicity of 2 Ethyl-Carbamates with Activity against Rhipicephalus microplus in Rats

    PubMed Central

    Prado-Ochoa, María Guadalupe; Gutiérrez-Amezquita, Ricardo Alfonso; Abrego-Reyes, Víctor Hugo; Velázquez-Sánchez, Ana María; Muñoz-Guzmán, Marco Antonio; Ramírez-Noguera, Patricia; Angeles, Enrique; Alba-Hurtado, Fernando

    2014-01-01

    The acute oral and dermal toxicity of two new ethyl-carbamates (ethyl-4-bromophenyl-carbamate and ethyl-4-chlorophenyl-carbamate) with ixodicide activity was determined in rats. The oral LD50 of each carbamate was 300 to 2000 mg/kg, and the dermal LD50 of each carbamate was >5000 mg/kg. Clinically, the surviving rats that had received oral doses of each carbamate showed decreased weight gain (P < 0.05) and had slight nervous system manifestations. These clinical signs were evident from the 300 mg/kg dose and were reversible, whereas the 2000 mg/kg dose caused severe damage and either caused their death or was motive for euthanasia. At necropsy, these rats had dilated stomachs and cecums with diffuse congestion, as well as moderate congestion of the liver. Histologically, the liver showed slight degenerative lesions, binucleated hepatocytes, focal coagulative necrosis, and congestion areas; the severity of the lesions increased with dosage. Furthermore, an slight increase in gamma-glutamyltransferase, lactate dehydrogenase, and creatinine was observed in the plasma. The dermal application of the maximum dose (5000 mg/kg) of each carbamate did not cause clinical manifestations or liver and skin alterations. This finding demonstrates that the carbamates under study have a low oral hazard and low acute dermal toxicity. PMID:24883331

  15. Immunogenic potential of the recombinant Rhipicephalus microplus aquaporin protein against the tick Rhipicephalus sanguineus Latreille, 1806 in domestic dogs

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aquaporins regulate water transport through the highly hydrophobic lipid bilayer of cell membranes. As ticks ingest large volumes of host blood in relation to their size, they are required to concentrate blood components and have efficient water transport mechanisms. This study aimed to evaluate the...

  16. Assessment of acute oral and dermal toxicity of 2 ethyl-carbamates with activity against Rhipicephalus microplus in rats.

    PubMed

    Prado-Ochoa, María Guadalupe; Gutiérrez-Amezquita, Ricardo Alfonso; Abrego-Reyes, Víctor Hugo; Velázquez-Sánchez, Ana María; Muñoz-Guzmán, Marco Antonio; Ramírez-Noguera, Patricia; Angeles, Enrique; Alba-Hurtado, Fernando

    2014-01-01

    The acute oral and dermal toxicity of two new ethyl-carbamates (ethyl-4-bromophenyl-carbamate and ethyl-4-chlorophenyl-carbamate) with ixodicide activity was determined in rats. The oral LD50 of each carbamate was 300 to 2000 mg/kg, and the dermal LD50 of each carbamate was >5000 mg/kg. Clinically, the surviving rats that had received oral doses of each carbamate showed decreased weight gain (P < 0.05) and had slight nervous system manifestations. These clinical signs were evident from the 300 mg/kg dose and were reversible, whereas the 2000 mg/kg dose caused severe damage and either caused their death or was motive for euthanasia. At necropsy, these rats had dilated stomachs and cecums with diffuse congestion, as well as moderate congestion of the liver. Histologically, the liver showed slight degenerative lesions, binucleated hepatocytes, focal coagulative necrosis, and congestion areas; the severity of the lesions increased with dosage. Furthermore, an slight increase in gamma-glutamyltransferase, lactate dehydrogenase, and creatinine was observed in the plasma. The dermal application of the maximum dose (5000 mg/kg) of each carbamate did not cause clinical manifestations or liver and skin alterations. This finding demonstrates that the carbamates under study have a low oral hazard and low acute dermal toxicity.

  17. Peripheral cellular and humoral responses to infestation with the cattle tick Rhipicephalus microplus in Santa Gertrudis cattle.

    PubMed

    Piper, E K; Jonsson, N N; Gondro, C; Vance, M E; Lew-Tabor, A; Jackson, L A

    2017-01-01

    Resistance to cattle tick infestation in single-host ticks is primarily manifested against the larval stage and results in the immature tick failing to attach successfully and obtain a meal. This study was conducted to identify immune responses that characterize the tick-resistant phenotype in cattle. Thirty-five tick-naïve Santa Gertrudis heifers were used in this study, thirty of which were artificially infested for thirteen weeks with tick larvae while five animals remained at a tick-free quarantine property to serve as a control group. Following thirteen weeks of tick infestation, the animals in this trial exhibited highly divergent tick-resistant phenotypes. Blood samples collected throughout the trial were used to measure peripheral immune parameters: haematology, the percentage of cellular subsets comprising the peripheral blood mononuclear cell (PBMC) population, tick-specific IgG1 and IgG2 antibody titres, IgG1 avidity for tick antigens and the ability of PBMC to recognize and proliferate in response to stimulation with tick antigens in vitro. The tick-susceptible cattle developed significantly higher tick-specific IgG1 antibody titres compared to the tick-resistant animals. These results suggest that the heightened antibody response either does not play a role in resistance or might contribute to increased susceptibility to infestation.

  18. Ticks (Acari: Ixodida) on wild carnivores in Brazil.

    PubMed

    Labruna, Marcelo B; Jorge, Rodrigo S P; Sana, Dênis A; Jácomo, Anah Tereza A; Kashivakura, Cyntia K; Furtado, Mariana M; Ferro, Claudia; Perez, Samuel A; Silveira, Leandro; Santos, Tarcísio S; Marques, Samuel R; Morato, Ronaldo G; Nava, Alessandra; Adania, Cristina H; Teixeira, Rodrigo H F; Gomes, Albério A B; Conforti, Valéria A; Azevedo, Fernando C C; Prada, Cristiana S; Silva, Jean C R; Batista, Adriana F; Marvulo, Maria Fernanda V; Morato, Rose L G; Alho, Cleber J R; Pinter, Adriano; Ferreira, Patrícia M; Ferreira, Fernado; Barros-Battesti, Darci M

    2005-01-01

    species), Dermacentor (1 species), Rhipicephalus (1 species), and Boophilus (1 species). Additional 5 tick species (3 Amblyomma species and 1 species from each of the genera Ixodes and Ornithodoros) were reported in the literature. The most common ticks on Carnivora hosts were Amblyomma ovale (found on 14 host species), Amblyomma cajennense (10 species), Amblyomma aureolatum (10 species), Amblyomma tigrinum (7 species), Amblyomma parvum (7 species), and Boophilus microplus (7 species).

  19. Epizootiology of Babesia bovis and Babesia bigemina in free-ranging white-tailed deer in northeastern Mexico.

    PubMed

    Cantu-C, Antonio; Ortega-S, J Alfonso; García-Vázquez, Zeferino; Mosqueda, Juan; Henke, Scott E; George, John E

    2009-06-01

    Species of Rhipicephalus (Boophilus) ticks are the vectors of babesiosis (cattle fever tick), which are distributed worldwide. White-tailed deer (Odocoileus virginianus) are important secondary hosts for the cattle fever ticks, Rhipicephalus (B.) annulatus and Rhipicephalus (B.) microplus. White-tailed deer are capable of sustaining Boophilus spp. tick populations in the presence or absence of cattle. The objectives of this study were to determine the frequency of Babesia bovis and Babesia bigemina and the prevalence of antibodies to them and identify possible risk factors for bovine babesiosis in white-tailed deer in 3 northeastern states of México. Whole blood and serum samples (n = 457) were collected from white-tailed deer in the states of Coahuila, Nuevo Leon, and Tamaulipas during the spring of 2004. Samples were tested for B. bovis and B. bigemina by nested polymerase chain reaction (n-PCR) (the primers for B. bovis identified the gene Rap-1 and B. bigemina were specific primers) and by an indirect immunofluorescence antibody test (IFAT). A questionnaire was given to each ranch to obtain information about management practices. Logistic regression methods were used to test the association between management factors and the dependent variable of positive n-PCR or IFAT. Nineteen (4.2%) samples were positive to B. bigemina and 6 (1.7%) were positive to B. bovis by n-PCR. Serological testing showed 59.9% (n = 274) of deer sampled were positive to B. bovis and 5.4% (n = 25) were positive to B. bigemina antibodies. The logistic model varied with different dependent variables. With positive n-PCR and B. bigemina as the dependent variable, 3 factors were associated: habitat (presence of brush and exotic grasses; odds ratio (OR), 3.3; 95% confidence interval (CI), 1.3-8.5), grazing system (continuous grazing OR 4.0; CI, 1.3-12.2), and tick treatment frequency (3-4 mo; OR 7.0, CI 1.4-34.3; 5-6 mo; OR, 11.0; CI, 1.9-62.7; > 6 mo; OR, 4.6; CI, 0.9-23.3). These findings

  20. Efficacy of larvicidal activity of green synthesized titanium dioxide nanoparticles using Mangifera indica extract against blood-feeding parasites.

    PubMed

    Rajakumar, Govindasamy; Rahuman, Abdul Abdul; Roopan, Selvaraj Mohana; Chung, Ill-Min; Anbarasan, Karunanithi; Karthikeyan, Viswanathan

    2015-02-01

    Titanium dioxide nanoparticles (TiO2 NPs) are considered to be among the best photocatalytic materials due to their long-term thermodynamic stability, strong oxidizing power, and relative non-toxicity. Nano-preparations with TiO2 NPs are currently under investigation as novel treatments for acne vulgaris, recurrent condyloma acuminata, atopic dermatitis, hyperpigmented skin lesions, and other non-dermatologic diseases. The present study was to investigate the acaricidal and larvicidal activity of synthesized TiO2 NPs utilizing leaf aqueous extract of Mangifera indica L. (Anacardiaceae) against hematophagous parasites. The anti-parasitic activity of TiO2 NPs against the larvae of Rhipicephalus (Boophilus) microplus, Hyalomma anatolicum anatolicum and Haemaphysalis bispinosa (Acari: Ixodidae), fourth instar larvae of Anopheles subpictus, and Culex quinquefasciatus (Diptera: Culicidae) were assessed. The green synthesized TiO2 NPs were analyzed by UV-Vis, FTIR, X-ray diffraction (XRD), AFM, SEM, and TEM. The XRD analysis of synthesized TiO2 NPs revealed the dominant peak at 2θ value of 27.81 which matched the 110 crystallographic plane of the rutile structure indicating the crystal structure. The FTIR spectra exhibited a prominent peak at 3,448 cm(-1) and showed OH stretching due to the alcoholic group, and the OH group may act as a capping agent. The SEM images of TiO2 NPs displayed spherical, oval in shape, individual, and some in aggregates. Characterization of the synthesized TiO2 NPs using AFM offered three-dimensional visualization and uneven surface morphology. The TEM micrograph showed agglomerates, round and slight elongation with an average size of 30 ± 5 nm. The maximum efficacy was observed in synthesized TiO2 NPs against the larvae of R. microplus, Hyalomma anatolicum anatolicum, Haemaphysalis bispinosa, A. subpictus, and Culex quinquefasciatus with LC50 value of 28.56, 33.17, 23.81, 5.84, and 4.34 mg/L, respectively. In the present study, a novel

  1. Entomopathogenic marine actinomycetes as potential and low-cost biocontrol agents against bloodsucking arthropods.

    PubMed

    Loganathan, Karthik; Kumar, Gaurav; Kirthi, Arivarasan Vishnu; Rao, Kokati Venkata Bhaskara; Rahuman, Abdul Abdul

    2013-11-01

    A novel approach to control strategies for integrated blood-feeding parasite management is in high demand, including the use of biological control agents. The present study aims to determine the efficacy of optimized crude extract of actinomycetes strain LK1 as biological control agent against the fourth-instar larvae of Anopheles stephensi and Culex tritaeniorhynchus (Diptera: Culicidae) and adults of Haemaphysalis bispinosa, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae), and Hippobosca maculata (Diptera: Hippoboscidae). Antiparasitic activity was optimized using the Plackett-Burman method, and the design was developed using the software Design-Expert version 8.0.7.1. The production of the optimized crude actinomycetes LK1 strain extract was performed using response surface methodology to optimize the process parameters of protease inhibitor activity of marine actinobacteria for the independent variables like pH, temperature, glucose, casein, and NaCl at two levels (-1 and +1). The potential actinomycetes strain was identified as Saccharomonas spp., and the metamodeling surface simulation procedure was followed. It was studied using a computer-generated experimental design, automatic control of simulation experiments, and sequential optimization of the metamodels fitted to a simulation response surface function. The central composite design (CCD) used for the analysis of treatment showed that a second-order polynomial regression model was in good agreement with the experimental results at R (2) = 0.9829 (p < 0.05). The optimized values of the variables for antioxidant production were pH 6.00, glucose 1.3%, casein 0.09%, temperature 31.23 °C, and NaCl 0.10%. The LK1 strain-optimized crude extract was purified using reversed-phase high-pressure liquid chromatography, and the isolated protease inhibitor showed antiparasitic activity. The antiparasitic activity of optimized crude extract of LK1 was tested against larvae of A. stephensi (LC₅₀ = 31.82 ppm

  2. Endectocide activity of a pour-on formulation containing 1.5 per cent ivermectin +0.5 per cent abamectin in cattle

    PubMed Central

    Silva, Heloisa Cristina; Prette, Nancy; Lopes, Welber Daniel Zanetti; Sakamoto, Cláudio Alessandro M; Buzzulini, Carolina; dos Santos, Thais Rabelo; Cruz, Breno Cayeiro; Teixeira, Weslen F Pires; Felippelli, Gustavo; Carvalho, Rafael Silveira; Maciel, Willian Giquelin; Soares, Vando Edésio; da Costa, Alvimar José

    2015-01-01

    The present work aimed to evaluate, through ten different studies, the therapeutic efficacy of a new pour-on formulation, containing 1.5 per cent ivermectin +0.5 per cent abamectin, against parasites of cattle. Results obtained on trials against Rhipicephalus (Boophilus) microplus showed that the pour-on combination of 1.5 per cent ivermectin +0.5 per cent abamectin obtained superior efficacy indexes against this ectoparasite, when compared with formulations containing 0.5 per cent ivermectin, 1 per cent ivermectin and the combination of 1 per cent abamectin +20 per cent levamisole. The results of efficacy of the ivermectin+abamectin and the 0.5 per cent ivermectin against Haematobia irritans were similar. Against Cochliomyia hominivorax larvae, all pour-on formulations tested (1.5 per cent ivermectin +0.5 per cent abamectin, 0.5 per cent ivermectin and 0.5 per cent abamectin), as well as 1 per cent doramectin administered subcutaneously, were considered ineffective. Cattle medicated with 1.5 per cent ivermectin +0.5 per cent abamectin, pour-on, remained free from parasitism by Dermatobia hominis larvae during 42 days (96 per cent efficacy), while values superior to 90 per cent were obtained by 0.5 per cent ivermectin (92 per cent) and 0.5 per cent abamectin (93 per cent) until the 42nd and 35th days post treatment, respectively. Against Haemonchus placei and Oesophagostomum radiatum, the pour-on of ivermectin+abamectin showed better efficacy than the 0.5 per cent ivermectin and 0.5 per cent abamectin. As to Cooperia punctata, there was no difference regarding efficacy results obtained by the avermectins combination and abamectin. The pour-on combination of 1.5 per cent ivermectin +0.5 per cent abamectin obtained high efficacy against R. (B.) microplus, D. hominis and some species of cattle gastrointestinal helminths when compared with formulations of 0.5 per cent ivermectin and 0.5 per cent abamectin administered through the same route. PMID:26392893

  3. A novel Ehrlichia genotype strain distinguished by the TRP36 gene naturally infects cattle in Brazil and causes clinical manifestations associated with ehrlichiosis.

    PubMed

    Aguiar, Daniel M; Ziliani, Thayza F; Zhang, Xiaofeng; Melo, Andreia L T; Braga, Isis A; Witter, Rute; Freitas, Leodil C; Rondelli, André L H; Luis, Michele A; Sorte, Eveline C B; Jaune, Felipe W; Santarém, Vamilton A; Horta, Mauricio C; Pescador, Carolina A; Colodel, Edson M; Soares, Herbert S; Pacheco, Richard C; Onuma, Selma S M; Labruna, Marcelo B; McBride, Jere W

    2014-09-01

    A novel Ehrlichia genotype most closely related to E. canis was reported in North American cattle in 2010, and a similar agent was subsequently identified in the hemolymph of Brazilian Rhipicephalus (Boophilus) microplus ticks and isolated in 2012. The purpose of this study was to determine whether this or other novel ehrlichial agents naturally infect Brazilian cattle. Using PCR targeting the genus-conserved dsb gene, DNA from this novel ehrlichial agent in Brazilian cattle was detected. Attempts to isolate the organism in vitro were performed using DH82 cells, but morulae and ehrlichial DNA could only be detected for approximately one month. In order to further molecularly characterize the organism, PCR was performed using primers specific for multiple E. canis genes (dsb, rrs, and trp36). Sequence obtained from the conserved rrs and dsb genes demonstrated that the organism was 99-100% identical to the novel Ehrlichia genotypes previously reported in North American cattle (rrs gene) and Brazilian ticks (rrs and dsb genes). However, analysis of the trp36 gene revealed substantial strain diversity between these Ehrlichia genotypes strains, including divergent tandem repeat sequences. In order to obtain preliminary information on the potential pathogenicity of this ehrlichial agent and clinical course of infection, a calf was experimentally infected. The calf showed clinical signs of ehrlichiosis, including fever, depression, lethargy, thrombocytopenia, and morulae were observed in peripheral blood monocytes. This study reports a previously unrecognized disease-causing Ehrlichia sp. in Brazilian cattle that is consistent with the genotype previously described in North America cattle and ticks from Brazil. Hence, it is likely that this is the organism previously identified as Ehrlichia bovis in Brazil in 1982. Furthermore, we have concluded that strains of these Ehrlichia genotypes can be molecularly distinguished by the trp36 gene, which has been widely utilized to

  4. Molecular cloning and functional characterization of an aspartic protease from the hard tick Haemaphysalis longicornis.

    PubMed

    Boldbaatar, Damdinsuren; Sikalizyo Sikasunge, Chummy; Battsetseg, Badgar; Xuan, Xuenan; Fujisaki, Kozo

    2006-01-01

    Haemaphysalis longicornis cDNA encoding an aspartic protease (longepsin) was identified from a midgut cDNA library. The longepsin cDNA contains 1176bp that code for 392 amino acid residues with a predictable molecular weight of 39.3kDa. The cDNA has a signal peptide sequence associated with the N-terminal domains and domain structure analysis revealed that the deduced protein has two aspartic acid residues that are characteristic of a single active site for aspartic proteases. This novel longepsin cDNA exhibits 57% identity to the lysosomal aspartic protease of Aedes aegypti, 52% to Bombyx mori cathepsin D, 38% to Ancylostoma caninum, 44% to Schistosoma mansoni and 28% to Boophilus microplus aspartic proteases. The DNA fragment coding for longepsin was cloned into a pGEX-4T-3 vector and expressed in Escherichia coli. The recombinant longepsin, once activated was able to hydrolyze casein substrate as well as hemoglobin (Hb) under acidic conditions (pH 3.5). RT-PCR analysis showed that the longepsin mRNA transcripts were expressed in salivary glands and midgut and not in the ovary. Northern blot analysis revealed that longepsin (1.5kb) was expressed in unfed and partially fed ticks and expression levels increased during feeding. The finding that longepsin is expressed in the midgut and salivary glands, proteolytic activity occurs under acidic conditions and longepsin can be gene silenced of longepsin provides compelling support for the hypothesis that longepsin plays an integral role in the proteolysis of erythrocyte Hb obtained from a host blood meal.

  5. The molecular and biological analysis of ixodid ticks histamine release factors.

    PubMed

    Mulenga, Albert; Azad, Abdu F

    2005-01-01

    We previously described a Dermacentor varibialis (DV) cDNA that encodes a ubiquitously expressed and tick saliva-secreted functional histamine release factor (HRF) homolog. In this study gene specific primers based on DVHRF open reading frame nucleotide sequence were utilized to amplify three orthologs, from the wood tick, D. andersoni (DA), the black legged tick, the southern cattle tick, Boophilus microplus (BM) and the lone star tick, Amblyomma americanum (AA). At nucleotide level, sequence comparisons revealed 98 89 and 84% similarity to DVHRF for DAHRF, AAHRF and BMHRF, respectively, while predicted polypeptide comparisons revealed 98, 96 and 91% similarity for DAHRF, AAHRF and BMHRF respectively. Phylogenetically, the tick HRF clade, while distinct (100% bootstrap value), is closely related to other arthropods, but distantly related to vertebrate and protozoan clades. Consistent with sequence similarity analysis, a DVHRF-specific northern blotting probe hybridized a approximately 900 base pair (bp) mRNA band on all RNA blots. Likewise a mouse polyclonal antibody to E. coli-expressed recombinant (r) DVHRF, cross-reacted baculovirus-expressed non-fusion rAAHRF, rDAHRF, and rBMHRF. As revealed by northern blotting analysis of larvae and nymph RNA, DVHRF mRNA is expressed in both immature and mature ticks indicating that its transcription is not developmentally regulated. Unlike rHRF/TCTP proteins of other organisms, the calcium-binding function may not be conserved for tick HRF homologs as revealed by the 45CaCl2+ overlay assay. Apparent global expression of DVHRF and its orthologs make this protein family an ideal target antigen for development of novel tick control strategies targeting multiple tick species.

  6. Hepatozoon and Theileria species detected in ticks collected from mammals and snakes in Thailand.

    PubMed

    Sumrandee, Chalao; Baimai, Visut; Trinachartvanit, Wachareeporn; Ahantarig, Arunee

    2015-04-01

    We report the detection of Hepatozoon and Theileria in 103 ticks from mammals and snakes in Thailand. By using a genus-specific 18S rRNA PCR, Hepatozoon and Theileria spp. were detected in 8% and 18%, respectively, of ticks (n=79) removed from mammals. Of the ticks removed from snakes (n=24), 96% were infected with Hepatozoon spp., but none were infected with Theileria. Phylogenetic analysis revealed that Hepatozoon spp. detected from Dermacentor astrosignatus and Dermacentor auratus ticks from Wild boar (Sus scrofa) formed a phylogenetic group with many isolates of Hepatozoon felis that were distantly related to a species group containing Hepatozoon canis and Hepatozoon americanum. In contrast, a phylogenetic analysis of the Hepatozoon sequences of snake ticks revealed that Hepatozoon spp. from Amblyomma varanense from King cobra (Ophiophagus hannah) and Amblyomma helvolum ticks from Indochinese rat snake (Ptyas korros), and Asiatic water snake (Xenochrophis piscator) are grouped with Hepatozoon spp. recently isolated from Monocellate cobras, Reticulated pythons and Burmese pythons, all of Thai origin, and with Hepatozoon sp. 774c that has been detected from a tick species obtained from Argus monitors in Australia. A phylogenetic analysis demonstrated that Theileria spp. from Rhipicephalus (Boophilus) microplus, Haemaphysalis obesa, and Haemaphysalis lagrangei ticks from Sambar deer (Cervus unicolor) cluster with the Theileria cervi isolates WU11 and 239, and Theileria sp. Iwate 141. We report for the first time a Hepatozoon species that shares genetic similarity with Hepatozoon felis found in Dermacentor astrosignatus and Dermacentor auratus ticks collected from Wild boars in Thailand. In addition, we found the presence of a Theileria cervi-like sp. which suggests the potential role of Haemaphysalis lagrangei as a Theileria vector in Thailand.

  7. Detection of Ehrlichia spp., Anaplasma spp., Rickettsia spp., and other eubacteria in ticks from the Thai-Myanmar border and Vietnam.

    PubMed

    Parola, Philippe; Cornet, Jean-Paul; Sanogo, Yibayiri Osée; Miller, R Scott; Thien, Huynh Van; Gonzalez, Jean-Paul; Raoult, Didier; Telford III, Sam R; Wongsrichanalai, Chansuda

    2003-04-01

    A total of 650 ticks, including 13 species from five genera, were collected from animals, from people, or by flagging of the vegetation at sites on the Thai-Myanmar border and in Vietnam. They were tested by PCR to detect DNA of bacteria of the order RICKETTSIALES: Three Anaplasma spp. were detected in ticks collected in Thailand, including (i) Anaplasma sp. strain AnDa465, which was considered a genotype of Anaplasma platys (formerly Ehrlichia platys) and which was obtained from Dermacentor auratus ticks collected from dogs; (ii) Anaplasma sp. strain AnAj360, which was obtained from Amblyomma javanense ticks collected on a pangolin; and (iii) Anaplasma sp. strain AnHl446, which was closely related to Anaplasma bovis and which was detected in Haemaphysalis lagrangei ticks collected from a bear. Three Ehrlichia spp. were identified, including (i) Ehrlichia sp. strain EBm52, which was obtained from Boophilus microplus ticks collected from cattle from Thailand; (ii) Ehrlichia sp. strain EHh324, which was closely related to Ehrlichia chaffeensis and which was detected in Haemaphysalis hystricis ticks collected from wild pigs in Vietnam; and (iii) Ehrlichia sp. strain EHh317, which was closely related to Ehrlichia sp. strain EBm52 and which was also detected in H. hystricis ticks collected from wild pigs in Vietnam. Two Rickettsia spp. were detected in Thailand, including (i) Rickettsia sp. strain RDla420, which was detected in Dermacentor auratus ticks collected from a bear, and (ii) Rickettsia sp. strain RDla440, which was identified from two pools of Dermacentor larvae collected from a wild pig nest. Finally, two bacteria named Eubacterium sp. strain Hw124 and Eubacterium sp. strain Hw191 were identified in Haemaphysalis wellingtoni ticks collected from chicken in Thailand; these strains could belong to a new group of bacteria.

  8. Cattle experimentally infected by Anaplasma marginale: Influence of splenectomy on disease pathogenesis, oxidative profile, and antioxidant status.

    PubMed

    Doyle, Rovaina L; França, Raqueli T; Oliveira, Camila B; Rezer, João F P; Klafke, Guilherme M; Martins, João R; Santos, Andrea P; do Nascimento, Naíla C; Mesick, Joanne B; Lopes, Sonia T A; Leal, Daniela B R; Da Silva, Aleksandro S; Andrade, Cinthia M

    2016-06-01

    Bovine anaplasmosis is caused by the obligate intraerythrocytic bacteria Anaplasma marginale. These bacteria are transmitted by tick species such as Rhipicephalus (Boophilus) microplus, blood-sucking insects, and fomites (needles, clippers, and other blood contaminated equipment). During the acute phase of infection, animals may develop fever, anemia, jaundice, and hepatosplenomegaly. The aims of this study are to quantify the bacteremia by quantitative PCR in eight naïve calves experimentally infected by A. marginale [splenectomized (n = 4), and intact/non-splenectomized (n = 4)], and to correlate these findings with markers of oxidative stress on days 0, 8, 15, 21 and 23 post-infection. Complete blood counts (CBC) were performed in both groups. Lipid peroxidation was estimated by quantifying thiobarbituric acid reactive substances (TBARS); and non-enzymatic antioxidants were assessed by erythrocyte content of non-protein thiols (NPSH). There were no significant differences in complete blood counts (CBC) between the two groups. However, both groups had a slight decrease on packet cell volume (PCV), erythrocytes and hemoglobin concentration, as well as an increase in total leukocyte counts due to elevated lymphocytes when comparing pre and post-infection with A. marginale. Progressive increase on TBARS levels and concomitant decrease on NPSH content were observed in all animals, without significant differences between splenectomized and intact animals. A positive correlation between bacteremia and TBARS, and a negative correlation between bacteremia and NPSH were observed in both groups with higher correlation for NPSH in splenectomized animals. A negative correlation between TBARS and NPSH levels was observed in both groups indicating lipid peroxidation without a non-enzymatic antioxidant response. The results of experimental infection by A. marginale in cattle showed that bacteremia has an impact on lipid peroxidation regardless of the splenectomy.

  9. Tick-borne bacteria in free-living jaguars (Panthera onca) in Pantanal, Brazil.

    PubMed

    Widmer, Cynthia E; Azevedo, Fernando C C; Almeida, Aliny P; Ferreira, Fernando; Labruna, Marcelo B

    2011-08-01

    Tick-borne bacteria were investigated in 10 free-living jaguars and their ticks in the Pantanal biome, Brazil. Jaguar sera were tested by indirect fluorescent antibody assays using Rickettsia rickettsii, Rickettsia parkeri, Rickettsia amblyommii, Rickettsia rhipicephali, Rickettsia felis, Rickettsia bellii, Ehrlichia canis, and Coxiella burnetii as crude antigens. All 10 jaguar sera reacted (titer ≥ 64) to at least one Rickettsia species; 4 and 3 sera reacted with E. canis and C. burnetii, respectively. One jaguar presented antibody titer to R. parkeri at least fourfold higher than those to any of the other five Rickettsia antigens, suggesting that this animal was infected by R. parkeri. Ticks collected from jaguars included the species Amblyomma cajennense, Amblyomma triste, and Rhipicephalus (Boophilus) microplus. No Rickettsia DNA was detected in jaguar blood samples, but an A. triste specimen collected on a jaguar was shown by PCR to be infected by R. parkeri. The blood of two jaguars and samples of A. triste, A. cajennense, and Amblyomma sp. yielded Ehrlichia DNA by PCR targeting the ehrlichial genes 16S rRNA and dsb. Partial DNA sequences obtained from PCR products resulted in a new ehrlichial strain, here designated as Ehrlichia sp. strain Jaguar. A partial DNA sequence of the 16S rRNA gene of this novel strain showed to be closest (99.0%) to uncultured strains of Ehrlichia sp. from Japan and Russia and 98.7% identical to different strains of Ehrlichia ruminantium. The ehrlichial dsb partial sequence of strain jaguar showed to be at most 80.7% identical to any Ehrlichia species or genotype available in GenBank. Through phylogenetic analysis, Ehrlichia sp. strain jaguar grouped in a cluster, albeit distantly, with different genotypes of E. ruminantium. Results highlight risks for human and animal health, considering that cattle ranching and ecotourism are major economic activities in the Pantanal region of Brazil.

  10. Diagnosis and management of bovine babesiosis outbreaks in cattle in Punjab state

    PubMed Central

    Bal, Mandeep Singh; Mahajan, Vishal; Filia, Gursimran; Kaur, Paramjit; Singh, Amarjit

    2016-01-01

    Aim: The aim of the present study was to diagnose severe outbreaks of bovine babesiosis in Punjab state, in the year 2015 and to suggest control and preventive measures to animal owners. Materials and Methods: Mortality of animals was recorded in two cattle herd comprising a total of 465 cattle in Sangrur (n=125) and Faridkot (n=340) districts. There was a history of purchase of animals at one farm. 23 blood samples were collected from diseased (n=15) and healthy animals (n=8) for hematological analysis, parasitological, and polymerase chain reaction (PCR)-based diagnosis. Ticks were also collected from animals for identification. Results: Out of 465 cattle at risk, 28 were critically ill and 14 died of disease with morbidity, mortality, and case fatality rate of 6.02%, 3.01%, and 50.00%, respectively. Clinical signs and necropsy findings were suggestive of babesiosis. Ticks collected from both the outbreaks were identified as Rhipicephalus (Boophilus) microplus. Thin blood smears from infected animals (especially with clinical sign of hemoglobinuria) were found positive for Babesia bigemina organisms; however, molecular diagnosis (PCR) further confirmed the disease. Animals were successfully treated with diminazene aceturate, hematinics, and antipyretics. Conclusions: Two fatal outbreaks of babesiosis in cattle were diagnosed with application of conventional parasitological, hematological, and molecular diagnostic techniques. PCR was found to be far more sensitive in detecting the disease, especially in latent infections. Animal owners were advised to follow quarantine measures before mixing new animals in the herd and strategic acaricidal treatments for effective tick control. PMID:28096607

  11. Molecular detection of Rickettsia, Anaplasma, Coxiella and Francisella bacteria in ticks collected from Artiodactyla in Thailand.

    PubMed

    Sumrandee, Chalao; Baimai, Visut; Trinachartvanit, Wachareeporn; Ahantarig, Arunee

    2016-07-01

    A total of 79 ticks collected from Sambar deer (Cervus unicolor), Barking deer (Muntiacus muntjak) and Wild boar (Sus scrofa) were examined by PCR for the presence of Rickettsia, Anaplasma, Coxiella, and Francisella bacteria. Of the 79 ticks, 13% tested positive for Rickettsia, 15% tested positive for Anaplasma, 4% tested positive for Coxiella, and 3% tested positive for Francisella. Interestingly, triple infection with Anaplasma, Rickettsia and Francisella was determined in a Dermacentor auratus tick. Moreover, another triple infection with Rickettsia, Anaplasma, and Coxiella was found in a Haemaphysalis lagrangei tick. Double infection of Rickettsia with Coxiella was also detected in another H. lagrangei tick. From the phylogenetic analyses, we found a Rickettsia sp. with a close evolutionary relationship to Rickettsia bellii in the H. lagrangei tick. We also found the first evidence of a Rickettsia sp. that is closely related to Rickettsia tamurae in Rhipicephalus (Boophilus) microplus ticks from Thailand. H. lagrangei and Haemaphysalis obesa ticks collected from Sambar deer tested positive for Anaplasma species form the same clade with Anaplasma bovis. In contrast, other H. lagrangei ticks collected from Sambar deer and D. auratus ticks collected from Wild boar were also reported for the first time to be infected with an Anaplasma species that is closely related to Anaplasma platys. The phylogenetic analysis of the 16S rRNA gene of Coxiella bacteria revealed that Coxiella symbionts from H. lagrangei formed a distinctly different lineage from Coxiella burnetii (a human pathogen). Additionally, Francisella bacteria identified in D. auratus ticks were found to be distantly related to a group of pathogenic Francisella species. The identification of these bacteria in several feeding ticks suggests the risk of various emerging tick-borne diseases and endosymbionts in humans, wildlife, and domestic animals in Thailand.

  12. Acute Oral Mammalian Toxicity and Effect of Solvents on Efficacy of Maerua edulis (Gilg. & Ben.) De Wolf against Rhipicephalus (Boophilus) decoloratus Koch, 1844 (Acarina: Ixodidae), Tick Larvae

    PubMed Central

    Mvumi, Brighton M.; Maramba, Tatenda

    2016-01-01

    Efficacy and toxicity of aqueous and organic solvents extracts of Maerua edulis against ticks and mice, respectively, were determined. Ground leaves were extracted separately using cold water, cold water plus surfactant (1% v/v liquid soap), hot water plus surfactant, hexane, or methanol to make 25% w/v stock solutions from which serial dilutions of 5, 10, 20, and 25% were made. For each concentration, 20 Rhipicephalus decoloratus tick larvae were put in filter papers impregnated with extracts and incubated for 48 h at 27°C and 85–90% RH for mortality observation after 24 h and 48 h. In the toxicity experiment, hot water plus surfactant treatments of 5, 10, 20, and 25% (w/v) M. edulis were administered in suspension per os to sexually mature Balb/C mice and observed for clinical signs and mortality for 72 h. Larvae mortality was highest (>98%) in methanol-extracted M. edulis treatments (20 and 25%), which was not different from the amitraz-based control (Tickbuster®). Mortality was also higher in the hot water than in cold water plus surfactant treatments (P < 0.05). No postadministration adverse health effects were observed in the mice. These results suggest that M. edulis is an effective tick remedy best extracted using methanol or hot water plus surfactant. PMID:27660762

  13. In Vitro Mode of Action and Anti-thrombotic Activity of Boophilin, a Multifunctional Kunitz Protease Inhibitor from the Midgut of a Tick Vector of Babesiosis, Rhipicephalus microplus

    PubMed Central

    Assumpção, Teresa C.; Ma, Dongying; Mizurini, Daniella M.; Kini, R. Manjunatha; Ribeiro, José M. C.; Kotsyfakis, Michail; Monteiro, Robson Q.; Francischetti, Ivo M. B.

    2016-01-01

    Background Hematophagous mosquitos and ticks avoid host hemostatic system through expression of enzyme inhibitors targeting proteolytic reactions of the coagulation and complement cascades. While most inhibitors characterized to date were found in the salivary glands, relatively few others have been identified in the midgut. Among those, Boophilin is a 2-Kunitz multifunctional inhibitor targeting thrombin, elastase, and kallikrein. However, the kinetics of Boophilin interaction with these enzymes, how it modulates platelet function, and whether it inhibits thrombosis in vivo have not been determined. Methodology/Principal Findings Boophilin was expressed in HEK293 cells and purified to homogeneity. Using amidolytic assays and surface plasmon resonance experiments, we have demonstrated that Boophilin behaves as a classical, non-competitive inhibitor of thrombin with respect to small chromogenic substrates by a mechanism dependent on both exosite-1 and catalytic site. Inhibition is accompanied by blockade of platelet aggregation, fibrin formation, and clot-bound thrombin in vitro. Notably, we also identified Boophilin as a non-competitive inhibitor of FXIa, preventing FIX activation. In addition, Boophilin inhibits kallikrein activity and the reciprocal activation, indicating that it targets the contact pathway. Furthermore, Boophilin abrogates cathepsin G- and plasmin-induced platelet aggregation and partially affects elastase-mediated cleavage of Tissue Factor Pathway Inhibitor (TFPI). Finally, Boophilin inhibits carotid artery occlusion in vivo triggered by FeCl3, and promotes bleeding according to the mice tail transection method. Conclusion/Significance Through inhibition of several enzymes involved in proteolytic cascades and cell activation, Boophilin plays a major role in keeping the midgut microenvironment at low hemostatic and inflammatory tonus. This response allows ticks to successfully digest a blood meal which is critical for metabolism and egg development. Boophilin is the first tick midgut FXIa anticoagulant also found to inhibit thrombosis. PMID:26745503

  14. Factors that influence the prevalence of acaricide resistance and tick-borne diseases.

    PubMed

    Foil, L D; Coleman, P; Eisler, M; Fragoso-Sanchez, H; Garcia-Vazquez, Z; Guerrero, F D; Jonsson, N N; Langstaff, I G; Li, A Y; Machila, N; Miller, R J; Morton, J; Pruett, J H; Torr, S

    2004-10-28

    This manuscript provides a summary of the results presented at a symposium organized to accumulate information on factors that influence the prevalence of acaricide resistance and tick-borne diseases. This symposium was part of the 19th International Conference of the World Association for the Advancement of Veterinary Parasitology (WAAVP), held in New Orleans, LA, USA, during August 10-14, 2003. Populations of southern cattle ticks, Boophilus microplus, from Mexico have developed resistance to many classes of acaricide including chlorinated hydrocarbons (DDT), pyrethroids, organophosphates, and formamidines (amitraz). Target site mutations are the most common resistance mechanism observed, but there are examples of metabolic mechanisms. In many pyrethroid resistant strains, a single target site mutation on the Na(+) channel confers very high resistance (resistance ratios: >1000x) to both DDT and all pyrethroid acaricides. Acetylcholine esterase affinity for OPs is changed in resistant tick populations. A second mechanism of OP resistance is linked to cytochrome P450 monooxygenase activity. A PCR-based assay to detect a specific sodium channel gene mutation that is associated with resistance to permethrin has been developed. This assay can be performed on individual ticks at any life stage with results available in a few hours. A number of Mexican strains of B. microplus with varying profiles of pesticide resistance have been genotyped using this test. Additionally, a specific metabolic esterase with permethrin-hydrolyzing activity, CzEst9, has been purified and its gene coding region cloned. This esterase has been associated with high resistance to permethrin in one Mexican tick population. Work is continuing to clone specific acetylcholinesterase (AChE) and carboxylesterase genes that appear to be involved in resistance to organophosphates. Our ultimate goal is the design of a battery of DNA- or ELISA-based assays capable of rapidly genotyping individual ticks to

  15. Tick repellents and acaricides of botanical origin: a green roadmap to control tick-borne diseases?

    PubMed

    Benelli, Giovanni; Pavela, Roman; Canale, Angelo; Mehlhorn, Heinz

    2016-07-01

    Arthropods are dangerous vectors of agents of deadly diseases, which may hit as epidemics or pandemics in the increasing world population of humans and animals. Among them, ticks transmit more pathogen species than any other group of blood-feeding arthropods worldwide. Thus, the effective and eco-friendly control of tick vectors in a constantly changing environment is a crucial challenge. A number of novel routes have been attempted to prevent and control tick-borne diseases, including the development of (i) vaccines against viruses vectored by ticks; (ii) pheromone-based control tools, with special reference to the "lure and kill" techniques; (iii) biological control programmes relying on ticks' natural enemies and pathogens; and (iv) the integrated pest management practices aimed at reducing tick interactions with livestock. However, the extensive employment of acaricides and tick repellents still remains the two most effective and ready-to-use strategies. Unfortunately, the first one is limited by the rapid development of resistance in ticks, as well as by serious environmental concerns. On the other hand, the exploitation of plants as sources of effective tick repellents is often promising. Here, we reviewed current knowledge concerning the effectiveness of plant extracts as acaricides or repellents against tick vectors of public health importance, with special reference to Ixodes ricinus, Ixodes persulcatus, Amblyomma cajennense, Haemaphysalis bispinosa, Haemaphysalis longicornis, Hyalomma anatolicum, Hyalomma marginatum rufipes, Rhipicephalus appendiculatus, Rhipicephalus (Boophilus) microplus, Rhipicephalus pulchellus, Rhipicephalus sanguineus and Rhipicephalus turanicus. Eighty-three plant species from 35 botanical families were selected. The most frequent botanical families exploited as sources of acaricides and repellents against ticks were Asteraceae (15 % of the selected studies), Fabaceae (9 %), Lamiaceae (10 %), Meliaceae (5 %), Solanaceae (6

  16. Poor inheritance of low attractiveness for Amblyomma variegatum in cattle.

    PubMed

    Stachurski, F

    2007-05-31

    Because of the high variability of cattle infestation, selective breeding has been proposed to control three-host ticks in Africa in the same way as it has been implemented to control the one-host tick Boophilus microplus in Australia. Interbreeding (divergent selection experiment) between Gudali zebu cattle was performed at the Wakwa Research Centre, Adamawa, Cameroon to assess the feasibility and impact of such a selective breeding programme. Fifty cows (chosen out of 129) and 2 bulls (chosen out of 10) were selected according to their greater or lesser attractiveness for the tick Amblyomma variegatum, assessed by their infestation degree (ID, ratio between individual animal infestation and mean herd infestation). Half of the animals displayed a high ID (H) and the remainder a low ID (L). The presence of a suckling calf had a significant effect on the infestation of its dam, the udder of lactating cows being significantly less infested (P<0.001) because of a tick removal effect by the calves. Observed infestation data were therefore corrected to allow comparison between lactating and non-lactating cows. The ID of the cows was assessed on five occasions from 1992 to 1994: the consistency of the infestation hierarchy was good overall (P<0.001) despite high variability seen in some animals which would complicate the selection of cattle with low infestation. Four breeding groups were set up (LxL, LxH, HxL, HxH) and 40 calves were born in 1993 and 1994. There was a correlation (P<0.02-0.001) between liveweight (or age) and ID when the younger calves of the groups were 6 months old, but the correlation disappeared by the time the calves were all older than 1 year. There was no difference between the average ID of the calves from the four breeding groups, and no correlation was observed between dam ID and calf ID. The average ID of the 10 calves born of the dams with the lowest ID was however significantly lower than that of the 10 calves born of the dams with the

  17. [Neurological complications of tick-borne relapsing fever].

    PubMed

    Castilla-Guerra, L; Fernandez-Moreno, M C; Vergara-Lopez, S; Merino-Rumin, M; Colmenero-Camacho, M A

    2016-09-16

    Introduccion. La fiebre recurrente transmitida por garrapatas (FRTG) puede producir complicaciones neurologicas. No existen apenas estudios en España sobre el tema. Objetivo. Estudiar la prevalencia y las caracteristicas clinicas de las complicaciones neurologicas de los pacientes con FRTG. Pacientes y metodos. Estudio retrospectivo de los pacientes atendidos con FRTG durante 12 años (2004-2015) en un hospital de una zona rural del sur de España. Resultados. Se incluyeron 75 pacientes, 42 varones (56%), con una edad media de 33 años (rango: 14-72 años). Se observaron picaduras de garrapatas en nueve pacientes (12%). Los sintomas mas frecuentes fueron: fiebre en 64 pacientes (85,3%), cefalea en 41 (54,6%) y vomitos en 26 (34,6%). Se sospecho afectacion meningea en nueve pacientes (12%), de los que tres (4%) tenian signos meningeos en el momento del ingreso. A todos ellos se les realizo una puncion lumbar. Ninguno presento paralisis facial ni otra manifestacion neurologica. Se encontraron alteraciones del liquido cefalorraquideo en los tres pacientes con meningismo. En uno de los casos se visualizo Borrelia en el liquido cefalorraquideo. En los pacientes con afectacion neurologica, el tratamiento utilizado fue penicilina G en un caso y ceftriaxona en dos. Todos los pacientes se recuperaron completamente. Conclusiones. La FRTG es una de las formas menos graves de borreliosis, y menos del 5% de los pacientes presenta complicaciones neurologicas. Sin embargo, los medicos deben saber que Borrelia puede causar meningitis en los sujetos expuestos a garrapatas en regiones endemicas de FRTG.

  18. The Genus Ixodes (Acari: Ixodidae) in Mexico: Adult Identification Keys, Diagnoses, Hosts, and Distribution (El genero Ixodes (Acari: Ixodidae) en Mexico: claves de identificacion para adultos, diagnosis, huespedes y distribucion)

    DTIC Science & Technology

    2010-01-01

    de Historia Natural 23:191-307. Hoffmann, A. 1969. Un caso de parálisis por picadura de garrapata. Revista Latinoamericana de Microbiología y...Hopkins University Press, Baltimore. p. 894-1531. Nuttall, G. H. F. 1916. Notes on ticks. IV. Relating to the genus Ixodes and including a description of...Ixodidae. Cambridge at the University Press, London. p. i-xix, 105-348. Robbins, R. G. and J. E. Keirans. 1992. Systematics and ecology of the

  19. Are white-tailed deer compromising efforts to maintain eradication of cattle fever ticks from cattle herds in the U.S.?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    From 1907 when the fever tick eradication campaign began until 1933, the two tick eradication methods of either dipping cattle in an acaricide or “pasture vacation” were enormously successful in eradicating cattle ticks [CT, Rhipicephalus (Boophilus) annulatus (Say)] and southern cattle ticks [SCT, ...

  20. Impact of white-tailed deer and exotic ungulate populations on cattle fever tick outbreaks in South Texas

    Technology Transfer Automated Retrieval System (TEKTRAN)

    From 1907 when the fever tick eradication campaign began until 1933, the two tick eradication methods of either dipping cattle in an acaricide or “pasture vacation” were enormously successful in eradicating cattle ticks [CT, Rhipicephalus (Boophilus) annulatus (Say)] and southern cattle ticks [SCT, ...

  1. Molecular Evidence for Anaplasma phagocytophilum in Israel

    PubMed Central

    Massung, Robert F.; Inbar, Moshe; Wallach, Arian D.; Shanas, Uri; Mumcuoglu, Kosta Y.; Waner, Trevor

    2007-01-01

    Sequences from the Anaplasma phagocytophilum 16S rRNA gene were detected in 5 ticks representing 3 species (Hyalomma marginatum, Rhipicephalus turanicus, and Boophilus kohlsi) collected from roe deer (Capreolus capreolus) in Mount Carmel, Israel. The sequences were all identical to those of Ap-variant 1 strain. PMID:18252125

  2. Transfected babesia bovis expressing a tick GST as a live vector vaccine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Rhipicephalus microplus tick is a notorious blood-feeding ectoparasite of livestock, especially cattle, responsible for massive losses in animal production. It is the main vector for transmission of pathogenic bacteria and parasites, including Babesia bovis, an intraerythrocytic apicomplexan pro...

  3. Rationale for classical biological control of cattle fever ticks and proposed methods for field collection of natural enemies

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Classical biological control using specialist parasitoids, predators and/or nematodes from the native ranges of cattle fever ticks Rhipicephalus microplus and Rhipicephalus annulatus could complement existing control strategies for this livestock pest in the transboundary region between Mexico and T...

  4. The US experience of eradicating and preventing re-infestation with cattle fever ticks

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The southern cattle tick, Rhipicephalus microplus, and the cattle tick, R. annulatus, are obligate ectoparasites and the vectors of pathogens causing bovine babesiosis, or “Cattle Fever”, and anaplasmosis in cattle. These cattle fever tick (CFT) species were declared eradicated from the U.S. in 1943...

  5. Collection of mitochondrial cytochrome oxidase I gene sequences from Rhipicephalus ticks from various geographic locations around the world

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Determining the origin of the cattle tick, Rhipicephalus microplus, will be helpful to the effort to find biological control agents. Molecular phylogenetics can assist in this determination. Thus, we sequenced and assembled partial gene sequences from the mitochondrial cytochrome oxidase I coding r...

  6. The novel isoxazoline ectoparasiticide fluralaner: selective inhibition of arthropod γ-aminobutyric acid- and L-glutamate-gated chloride channels and insecticidal/acaricidal activity.

    PubMed

    Gassel, Michael; Wolf, Christian; Noack, Sandra; Williams, Heike; Ilg, Thomas

    2014-02-01

    Isoxazolines are a novel class of parasiticides that are potent inhibitors of γ-aminobutyric acid (GABA)-gated chloride channels (GABACls) and L-glutamate-gated chloride channels (GluCls). In this study, the effects of the isoxazoline drug fluralaner on insect and acarid GABACl (RDL) and GluCl and its parasiticidal potency were investigated. We report the identification and cDNA cloning of Rhipicephalus (R.) microplus RDL and GluCl genes, and their functional expression in Xenopus laevis oocytes. The generation of six clonal HEK293 cell lines expressing Rhipicephalus microplus RDL and GluCl, Ctenocephalides felis RDL-A285 and RDL-S285, as well as Drosophila melanogaster RDLCl-A302 and RDL-S302, combined with the development of a membrane potential fluorescence dye assay allowed the comparison of ion channel inhibition by fluralaner with that of established insecticides addressing RDL and GluCl as targets. In these assays fluralaner was several orders of magnitude more potent than picrotoxinin and dieldrin, and performed 5-236 fold better than fipronil on the arthropod RDLs, while a rat GABACl remained unaffected. Comparative studies showed that R. microplus RDL is 52-fold more sensitive than R. microplus GluCl to fluralaner inhibition, confirming that the GABA-gated chloride channel is the primary target of this new parasiticide. In agreement with the superior RDL on-target activity, fluralaner outperformed dieldrin and fipronil in insecticidal screens on cat fleas (Ctenocephalides felis), yellow fever mosquito larvae (Aedes aegypti) and sheep blowfly larvae (Lucilia cuprina), as well as in acaricidal screens on cattle tick (R. microplus) adult females, brown dog tick (Rhipicephalus sanguineus) adult females and Ornithodoros moubata nymphs. These findings highlight the potential of fluralaner as a novel ectoparasiticide.

  7. Tick (Acari: Ixodidae) infestations in cattle along Geba River basin in Guinea-Bissau.

    PubMed

    Zúquete, Sara Tudela; Coelho, João; Rosa, Fernanda; Vaz, Yolanda; Cassamá, Bernardo; Padre, Ludovina; Santos, Dulce; Basto, Afonso P; Leitão, Alexandre

    2017-01-01

    Tick infestations are a major problem for animal production in tropical areas where prevention and control remain deficient. The present study sought to assess the awareness of traditional cattle producers towards the importance of ticks and aimed at the identification of tick species infesting bovines within the Geba River basin, Guinea-Bissau. Interviews with producers revealed that the majority directly correlates the presence of ticks with the occurrence of diseases in cattle. However, insufficient or inadequate control approaches prevail. A total of 337 ticks were collected on bovines at 18 different villages (10 during dry season, and 8 during rainy season). The tick species collected during the dry season were Rhipicephalus (Boophilus) geigyi (56.5%), followed by Amblyomma variegatum (23.3%), Rhipicephalus (Boophilus) annulatus (17.6%) and Hyalomma truncatum (1%). In the rainy season A. variegatum was the most collected (88.9%), followed by R. (Boophilus) geigyi (4.2%), R. (Boophilus) annulatus (3.4%), Rhipicephalus sanguineus group (2.8%) and H. truncatum (0.7%). To support species identification, segments of both cytochrome c oxidase I (COI) and 12S ribosomal RNA (12S) genes were sequenced and the data gathered were analysed by maximum likelihood and parsimony. Morphological and genetic data of individual specimens gathered in this study provide relevant information for future studies on tick population dynamics in the region. In addition, it led to a deeper characterization of R. sulcatus and a R. sanguineus-like specimen, exploring their genetic relationship with other R. sanguineus, which supports their classification as distinct species within R. sanguineus group.

  8. Identification of a CAPA-PVK (IXORI-PVK) from Single Cells of the Gulf Coast Tick, Amblyomma maculatum

    DTIC Science & Technology

    2008-01-01

    of the amino acid sequence of Ixori-PVK (PALIPFPRV-NH2), a periviscerokinin which had previously been identified from two other ticks, Ixodes ricinus...mass spectrometric single cell analysis. De novo sequencing yielded an amino acid sequence identical with the CAPA-PVK of Ixodes ricinus and Boophilus... manufacturer as: ‘none’, ‘medium’ and ‘high’. The fragmentation patterns from these three different settings were used to determine the sequence of the

  9. Molecular characterization of Anaplasma marginale in ticks naturally feeding on buffaloes.

    PubMed

    da Silva, Jenevaldo Barbosa; da Fonseca, Adivaldo Henrique; Barbosa, José Diomedes

    2015-10-01

    Anaplasma marginale is the most prevalent pathogen transmitted by ticks in cattle in tropical and subtropical regions of the world. However, the tick species involved in the transmission of A. marginale in buffaloes in Brazil have not been identified. The objective of the present study was to determine the presence of A. marginale in ticks parasitizing water buffaloes. A total of 200 samples of Rhipicephalus microplus, Dermacentor nitens, Amblyomma cajennense, and Amblyomma maculatum were collected and tested by conventional and quantitative PCR for the presence of the msp1a and msp5 genes. In the present study, 35 ticks (17.5%) were positive for A. marginale DNA by qPCR analysis. The positive ticks belonged to four different species: R. microplus (22.2%), A. cajennense (13.8%), A. maculatum (16.0%), and D. nitens (10.0%). Individuals of the three developmental stages (larvae, nymphs, and adults) of R. microplus and A. cajennense were found to be positive for A. marginale, only nymphs and adults of A. maculatum were found to be positive, and finally, only adults of D. nitens were positive for A. marginale. Our results suggest that R. microplus, A. cajennense, A. maculatum, and D. nitens ticks may be involved in the transmission of A. marginale in buffaloes. However, while A. marginale PCR positive ticks were recorded, this does not indicate vector competence; only that the ticks may contain a blood meal from an infected host. Additionally, the results show that the strains of A. marginale from buffaloes and cattle are phylogenetically related.

  10. Nellore cattle (Bos indicus) and ticks within the Brazilian Pantanal: ecological relationships.

    PubMed

    Ramos, Vanessa N; Piovezan, Ubiratan; Franco, Ana Helena A; Rodrigues, Vinicius S; Nava, Santiago; Szabó, Matias P J

    2016-02-01

    Pantanal is a huge floodplain mostly in Brazil, and its main economic activity is extensive cattle raising, in farms characterized by an extremely wildlife-rich environment. We herein describe tick infestations of cattle and of the natural environment in Pantanal of Nhecolândia in Brazil, at areas with and without cattle during both dry and wet seasons. Environmental sampling resulted in three tick species: Amblyomma sculptum (423 nymphs and 518 adults), Amblyomma parvum (7 nymphs and 129 adults), Amblyomma ovale (3 adults) as well as three clusters and two individuals of Amblyomma sp. larvae. A significantly higher number of adult A. sculptum ticks was found in areas with cattle in the wet season. From 106 examinations of bovines 1710 ticks from three species were collected: Rhipicephalus microplus (55.7% of the total), A. sculptum (38%) and A. parvum (4.1%), as well as 32 Amblyomma sp. larvae. A significant similarity was found between Amblyomma tick fauna from environment and on cattle during both seasons. All A. sculptum females on bovines were flat whereas many of A. parvum females and A. sculptum nymphs were engorging. Although R. microplus was the most abundant tick species on cattle, overall highest tick prevalence on bovines in the dry season was of A. sculptum nymphs. Lack of R. microplus in environmental sampling, relationship between cattle and increase in adult A. sculptum numbers in the environment as well as suitability of bovine for the various tick species are discussed.

  11. Low temperature affects cattle tick reproduction but does not lead to transovarial transmission of Anaplasma marginale.

    PubMed

    Esteves, E; Pohl, P C; Klafke, G M; Reck, J; Fogaça, A C; Martins, J R; Daffre, S

    2015-12-15

    Anaplasma marginale is an obligate intracellular pathogen that infects the erythrocytes of calves, causing bovine anaplasmosis. This rickettsia is biologically transmitted by several species of ticks. In tropical and subtropical regions of the world, Rhipicephalus microplus is the main vector. Due to their mobility and longevity, the adult males play an important role in the transmission of A. marginale to calves. Some studies have demonstrated that A. marginale can be intrastadially and interstadially transmitted in R. microplus, but the transovarial transmission has not been demonstrated so far. In the present study, we investigated the effects of low temperature on both the A. marginale migration from infected females to their offspring and reproductive parameters of the tick R. microplus. The larvae of R. microplus fed on a calf infected with the strain Jaboticabal of A. marginale. At the end of the parasitic phase, fully engorged females were incubated at either 18°C or 28°C for oviposition. Although A. marginale was detected in the salivary glands of the females, demonstrating that the ticks were successfully infected, the presence of rickettsia was not detected in the offspring. However, the preoviposition period of the non-infected females maintained at 18°C was longer than that of those maintained at 28°C. In addition, the average weight of the mass of eggs as well as the egg production efficiency (ratio of the egg mass weight to the female weight) of the females maintained at 18°C were significantly lower than those of the females incubated at 28°C. There was no larval hatching from the eggs maintained exclusively at 18°C, even at 65 days after female detachment. Hatching occurred only when the eggs maintained at 18°C were transferred to 28°C at 20 days after female detachment (18°C/28°C). We also verified a significantly higher larvae conversion efficiency (ratio of the larvae mass weight to the egg mass weight) in the group of females

  12. Rickettsia sp. strain colombianensi (Rickettsiales: Rickettsiaceae): a new proposed Rickettsia detected in Amblyomma dissimile (Acari: Ixodidae) from iguanas and free-living larvae ticks from vegetation.

    PubMed

    Miranda, Jorge; Portillo, Aránzazu; Oteo, José A; Mattar, Salim

    2012-07-01

    From January to December 2009, 55 Amblyomma dissimile (Koch) ticks removed from iguanas in the municipality of Monteria and 3,114 ticks [458 Amblyomma sp. larvae, 2,636 Rhipicephalus microplus (Canestrini) larvae and 20 Amblyomma sp. nymphs] collected over vegetation in Los Cordobas were included in the study. The ticks were pooled into groups from which DNA was extracted. For initial screening of Rickettsia sp., each pool was analyzed by gltA real-time polymerase chain reaction (PCR). Positive pools were further studied using gltA, ompA, and ompB conventional PCR assays. Sequencing and phylogenetic analysis were also conducted. Rickettsial DNA was found in 28 pools of ticks (16 A. dissimile pools and 12 free-living larvae pools) out of 113 (24.7%) using real-time PCR. The same 28 pools were also positive using conventional PCR assays aimed to amplify gltA, ompA, and ompB. For each gene analyzed, PCR products obtained from 4/28 pools (two pools of A. dissimile, one pool of Amblyomma sp. larvae and one pool of Rh. microplus larvae) were randomly chosen and sequenced twice. Nucleotide sequences generated were identical to each other for each of the rickettsial genes gltA, ompA, and ompB, and showed 99.4, 95.6, and 96.4% identity with those of Rickettsia tamurae. They were deposited in the GenBank database under accession numbers JF905456, JF905458, and JF905457, respectively. In conclusion, we present the first molecular evidence of a novel Rickettsia (Rickettsia sp. strain Colombianensi) infecting A. dissimile ticks collected from iguanas, and also Rh. microplus and unspeciated Amblyomma larvae from vegetation in Colombia.

  13. Ticks infesting humans in Northern Misiones, Argentina.

    PubMed

    Lamattina, Daniela; Nava, Santiago

    2016-01-01

    This work presents records of ticks infesting humans in northern Misiones Province, Argentina. Also, notes on potential transmission of tick-borne pathogens are included. A total of 282 ticks attached to researchers were collected and identified by their morphological characters. Eight tick species were found: Amblyomma brasiliense, Amblyomma coelebs, Amblyomma dubitatum, Amblyomma incisum, Amblyomma ovale, Haemaphysalis juxtakochi, Rhipicephalus sanguineus and Rhipicephalus microplus. Some of these species as A. dubitatum, A. ovale and R. sanguineus have been found infected with spotted fever group rickettsiae pathogenic to humans in Brazil and Argentina. The potential role as vectors of humans pathogens of the ticks found attached to humans in this study is discussed.

  14. A list of the 70 species of Australian ticks; diagnostic guides to and species accounts of Ixodes holocyclus (paralysis tick), Ixodes cornuatus (southern paralysis tick) and Rhipicephalus australis (Australian cattle tick); and consideration of the place of Australia in the evolution of ticks with comments on four controversial ideas.

    PubMed

    Barker, Stephen C; Walker, Alan R; Campelo, Dayana

    2014-10-15

    Seventy species of ticks are known from Australia: 14 soft ticks (family Argasidae) and 56 hard ticks (family Ixodidae). Sixteen of the 70 ticks in Australia may feed on humans and domestic animals (Barker and Walker 2014). The other 54 species of ticks in Australia feed only on wild mammals, reptiles and birds. At least 12 of the species of ticks in Australian also occur in Papua New Guinea. We use an image-matching system much like the image-matching systems of field guides to birds and flowers to identify Ixodes holocyclus (paralysis tick), Ixodes cornuatus (southern paralysis tick) and Rhipicephalus (Boophilus) australis (Australian cattle tick). Our species accounts have reviews of the literature on I. holocyclus (paralysis tick) from the first paper on the biology of an Australian tick by Bancroft (1884), on paralysis of dogs by I. holocyclus, to papers published recently, and of I. cornuatus (southern paralysis tick) and Rhipicephalus (Boophilus) australis (Australian cattle tick). We comment on four controversial questions in the evolutionary biology of ticks: (i) were labyrinthodont amphibians in Australia in the Devonian the first hosts of soft, hard and nuttalliellid ticks?; (ii) are the nuttalliellid ticks the sister-group to the hard ticks or the soft ticks?; (iii) is Nuttalliella namaqua the missing link between the soft and hard ticks?; and (iv) the evidence for a lineage of large bodied parasitiform mites (ticks plus the holothyrid mites plus the opiliocarid mites).

  15. Cattle tick vaccines: many candidate antigens, but will a commercially viable product emerge?

    PubMed

    Guerrero, Felix D; Miller, Robert J; Pérez de León, Adalberto A

    2012-05-01

    The cattle tick, Rhipicephalus microplus, is arguably the world's most economically important external parasite of cattle. Sustainable cattle tick control strategies are required to maximise the productivity of cattle in both large production operations and small family farms. Commercially available synthetic acaricides are commonly used in control and eradication programs, but indiscriminate practices in their application have resulted in the rapid evolution of resistance among populations in tropical and subtropical regions where the invasive R. microplus thrives. The need for novel technologies that could be used alone or in combination with commercially available synthetic acaricides is driving a resurgence of cattle tick vaccine discovery research efforts by various groups globally. The aim is to deliver a next-generation vaccine that has an improved efficacy profile over the existing Bm86-based cattle tick vaccine product. We present a short review of these projects and offer our opinion on what constitutes a good target antigen and vaccine, and what might influence the market success of candidate vaccines. The previous experience with Bm86-based vaccines offers perspective on marketing and producer acceptance aspects that a next-generation cattle tick vaccine product must meet for successful commercialisation.

  16. Hard ticks (Acari: Ixodidae) of livestock in Nicaragua, with notes about distribution.

    PubMed

    Düttmann, Christiane; Flores, Byron; Kadoch Z, Nathaniel; Bermúdez C, Sergio

    2016-09-01

    We document the species of ticks that parasitize livestock in Nicaragua. The study was based on tick collection on cattle and horses from 437 farms in nine departments. Of 4841 animals examined (4481 cows and 360 horses), 3299 were parasitized, which represent 68 % of the bovines and 67 % of the equines in study: 59 cows and 25 horses were parasitized by more than one species. In addition, 280 specimens of the entomological museum in León were examined. The ticks found on cattle were Rhipicephalus microplus (75.2 % of the ticks collected), Amblyomma mixtum (20.8 %), A. parvum (2.6 %), A. tenellum (0.7 %), A. maculatum (0.7 %). While the ticks collected from the horses were: Dermacentor nitens (41.5 %), A. mixtum (31.7 %), R. microplus (13.8 %), A. parvum (6.5 %), A. tenellum (3.3 %), D. dissimilis (2.4 %) and A. maculatum (0.8 %).

  17. Association of Anaplasma marginale Strain Superinfection with Infection Prevalence within Tropical Regions

    PubMed Central

    Castañeda-Ortiz, Elizabeth J.; Ueti, Massaro W.; Camacho-Nuez, Minerva; Mosqueda, Juan J.; Mousel, Michelle R.; Johnson, Wendell C.; Palmer, Guy H.

    2015-01-01

    Strain superinfection occurs when a second strain infects a host already infected with and having mounted an immune response to a primary strain. The incidence of superinfection with Anaplasma marginale, a tick-borne rickettsial pathogen of domestic and wild ruminants, has been shown to be higher in tropical versus temperate regions. This has been attributed to the higher prevalence of infection, with consequent immunity against primary strains and thus greater selective pressure for superinfection with antigenically distinct strains. However an alternative explanation would be the differences in the transmitting vector, Dermacentor andersoni in the studied temperate regions and Rhipicephalus microplus in the studied tropical regions. To address this question, we examined two tropical populations sharing the same vector, R. microplus, but with significantly different infection prevalence. Using two separate markers, msp1α (one allele per genome) and msp2 (multiple alleles per genome), there were higher levels of multiple strain infections in the high infection prevalence as compared to the low prevalence population. The association of higher strain diversity with infection prevalence supports the hypothesis that high levels of infection prevalence and consequent population immunity is the predominant driver of strain superinfection. PMID:25793966

  18. Strains of toxic and harmful microalgae, from waste water, marine, brackish and fresh water.

    PubMed

    Rodríguez-Palacio, M C; Crisóstomo-Vázquez, L; Alvarez-Hernández, S; Lozano-Ramírez, C

    2012-01-01

    Some microalgae are economically important in Mexico and the world because they can be potentially toxic. Algal explosive population growths are named harmful algal blooms and are frequently recorded in Mexico. The authors set up potentially toxic microalgae cultures from the Gulf of Mexico (Garrapatas tideland, Barberena river, Carpintero lagoon in Tamaulipas State; Chalchoapan and Catemaco lakes in Veracruz State), from the Mexican Pacific Ocean, Guerrero, Colima and Michoacán States, and from interior water bodies such as Vicente Aguirre dam, Chapultepec lake and several waste water treatment plants. This research is about the diversity and abundance of phytoplankton in relation a specific site because of harmful algal bloom events. Microalgae cultures are useful in order to solve taxonomic problems, to know life cycles, molecular studies, for the study of toxic species, and the isolation of useful metabolites. The cultures for this research are clonal, non-axenic, semi-continuous, 12:12 light/dark photoperiod, 20 ± 1 °C temperature and 90.5 µmol m(-2)s(-1) illumination. Four different culture media were used. This collection is open to the worldwide scientific community as a source of organisms in controlled conditions that can be used as a useful tool for microalgae research work.

  19. Transfected Babesia bovis Expressing a Tick GST as a Live Vector Vaccine

    PubMed Central

    Oldiges, Daiane P.; Laughery, Jacob M.; Tagliari, Nelson Junior; Leite Filho, Ronaldo Viana; Davis, William C.; da Silva Vaz, Itabajara; Termignoni, Carlos; Knowles, Donald P.; Suarez, Carlos E.

    2016-01-01

    The Rhipicephalus microplus tick is a notorious blood-feeding ectoparasite of livestock, especially cattle, responsible for massive losses in animal production. It is the main vector for transmission of pathogenic bacteria and parasites, including Babesia bovis, an intraerythrocytic apicomplexan protozoan parasite responsible for bovine Babesiosis. This study describes the development and testing of a live B. bovis vaccine expressing the protective tick antigen glutathione-S-transferase from Haemaphysalis longicornis (HlGST). The B. bovis S74-T3B parasites were electroporated with a plasmid containing the bidirectional Ef-1α (elongation factor 1 alpha) promoter of B. bovis controlling expression of two independent genes, the selectable marker GFP-BSD (green fluorescent protein–blasticidin deaminase), and HlGST fused to the MSA-1 (merozoite surface antigen 1) signal peptide from B. bovis. Electroporation followed by blasticidin selection resulted in the emergence of a mixed B. bovis transfected line (termed HlGST) in in vitro cultures, containing parasites with distinct patterns of insertion of both exogenous genes, either in or outside the Ef-1α locus. A B. bovis clonal line termed HlGST-Cln expressing intracellular GFP and HlGST in the surface of merozoites was then derived from the mixed parasite line HlGST using a fluorescent activated cell sorter. Two independent calf immunization trials were performed via intravenous inoculation of the HlGST-Cln and a previously described control consisting of an irrelevant transfected clonal line of B. bovis designated GFP-Cln. The control GFP-Cln line contains a copy of the GFP-BSD gene inserted into the Ef-1α locus of B. bovis in an identical fashion as the HIGST-Cln parasites. All animals inoculated with the HlGST-Cln and GFP-Cln transfected parasites developed mild babesiosis. Tick egg fertility and fully engorged female tick weight was reduced significantly in R. microplus feeding on HlGST-Cln-immunized calves

  20. Detection of haemoparasites in cattle by reverse line blot hybridisation with a note on the distribution of ticks in Sicily.

    PubMed

    Georges, K; Loria, G R; Riili, S; Greco, A; Caracappa, S; Jongejan, F; Sparagano, O

    2001-08-31

    A reverse line blot hybridisation (RLB) of 21 oligonucleotides with polymerase chain reaction (PCR) amplified regions of 16S rRNA (Ehrlichia/Anaplasma group) or 18S rRNA (Babesia/Theileria group) genes of haemoparasites detected Theileria annulata, T. buffeli/orientalis, Babesia bovis, B. bigemina, B. divergens, Ehrlichia bovis, Anaplasma marginale, A. centrale and unknown species within the Rickettsia tribe.A very high prevalence of mixed infections was detected, which indicated that animals infected with Babesia spp. were also infected with Theileria spp. and/or Anaplasma spp. The tick distribution appeared to be seasonal with Hyalomma marginatum as the most frequently observed tick and Boophilus annulatus and Ixodes ricinus as the least frequently observed ticks. Other species identified in the 818 ticks collected during the five sampling periods between April 1998 and November 1999 included H. lusitanicum, Rhipicephalus sanguineus group, R. bursa, Dermacentor marginatus, Haemaphysalis punctata, B. annulatus and I. ricinus.

  1. Rickettsia africae and Candidatus Rickettsia barbariae in Ticks in Israel

    PubMed Central

    Waner, Trevor; Keysary, Avi; Eremeeva, Marina E.; Din, Adi Beth; Mumcuoglu, Kosta Y.; King, Roni; Atiya-Nasagi, Yafit

    2014-01-01

    DNA of several spotted fever group rickettsiae was found in ticks in Israel. The findings include evidence for the existence of Rickettsia africae and Candidatus Rickettsia barbariae in ticks in Israel. The DNA of R. africae was detected in a Hyalomma detritum tick from a wild boar and DNA of C. Rickettsia barbariae was detected in Rhipicephalus turanicus and Rhipicephalus sanguineus collected from vegetation. The DNA of Rickettsia massiliae was found in Rh. sanguineus and Haemaphysalis erinacei, whereas DNA of Rickettsia sibirica mongolitimonae was detected in a Rhipicephalus (Boophilus) annulatus. Clinicians should be aware that diseases caused by a variety of rickettsiae previously thought to be present only in other countries outside of the Middle East may infect residents of Israel who have not necessarily traveled overseas. Furthermore, this study reveals again that the epidemiology of the spotted fever group rickettsiae may not only involve Rickettsia conorii but may include other rickettsiae. PMID:24615133

  2. Spotted fever group rickettsiae in ticks from the Masai Mara region of Kenya.

    PubMed

    Macaluso, Kevin R; Davis, Jon; Alam, Uzma; Korman, Amy; Rutherford, Jeremiah S; Rosenberg, Ronald; Azad, Abdu F

    2003-05-01

    We have identified for the first time Rickettsia africae, and the ticks that harbored them, in Kenya. A total of 5,325 ticks were collected from vegetation, livestock, and wild animals during two field trips to southwestern Kenya. Most were immature forms (85.2%) belonging to the genera Amblyomma or Rhipicephalus. The adults also included representatives from the genus Boophilus. Ticks were assessed for rickettsial DNA by a polymerase chain reaction (PCR) using primers for the spotted fever group (SFG)-specific rickettsial outer membrane protein A (rompA) gene, and positive amplicons were sequenced. While none of the immature ticks tested positive by PCR, 15.8% of the adult Amblyomma variegatum and less than 1% of the Rhipicephalus spp. were SFG positive. Sequences of amplified products were identified as R. africae. These findings extend the known range of R. africae.

  3. Ticks (Acari: Ixodidae) on wild animals from the Porto-Primavera Hydroelectric power station area, Brazil.

    PubMed

    Labruna, Marcelo B; de Paula, Cátia D; Lima, Thiago F; Sana, Dênis A

    2002-12-01

    From June 2000 to June 2001, a total of 741 ticks were collected from 51 free-living wild animals captured at the Porto-Primavera Hydroelectric power station area, located alongside an approximately 180 km course of the Paran river, between the states of S o Paulo and Mato Grosso do Sul, comprising 9 species of 3 genera: Ambly-omma (7 species), Boophilus (1) and Anocentor (1). A total of 421 immature Amblyomma ticks were reared in laboratory until the adult stage, allowing identification of the species. A. cajennense was the most frequent tick species (mostly immature stages) collected on 9 host species: Myrmecophaga tridactyla, Tamandua tetradactyla,Cerdocyon thous, Puma concolor,Tayassu tajacu, Mazama gouazoubira,Hydrochaeris hydrochaeris,Alouatta caraya, Cebus apella. Other tick species were less common, generally restricted to certain host taxa.

  4. Ticks status in Central Asia with a special emphasis on Uzbekistan.

    PubMed

    Rasulov, Ilham

    2007-09-01

    Uzbekistan is located between the greatest rivers of Central Asia and shares borders with Kazakhstan to the west and to the north, Kyrgyzstan and Tajikistan to the east, and Afghanistan and Turkmenistan to the south. The climate is severely continental and arid, with hot summers and cold winters. These climatic particularities of Uzbekistan determine the distribution of tick fauna. The Ixodidae family of ticks is represented by 23 species in Uzbekistan. These ticks, as ectoparasites, cause significant damage to the livestock breeding and also serve as carriers of many human and animal diseases. During the last 10 years, more than 30,000 ticks in different regions of Uzbekistan were collected and identified. Analysis showed that cattle are parasitized by 11 species of Ixodidae ticks. The dominating species were Hyalomma anatolicum (34.9%), Hyalomma detritum detritum (31.8%), Boophilus kohlsi (30.7%).

  5. Tick burden and prevalence of Theileria parva infection in Tarime zebu cattle in the lake zone of Tanzania.

    PubMed

    Laisser, Emmanuel Levillal Katamboi; Kipanyula, Maulilio John; Msalya, George; Mdegela, Robinson Hammerthon; Karimuribo, Esron Daniel; Mwilawa, Anjello Joseph; Mwega, Elisa Daniel; Kusiluka, Lughano; Chenyambuga, Sebastian Wilson

    2014-12-01

    This study was carried out to assess the distribution, abundance of different tick genera and prevalence of Theileria parva infection in Tarime zebu cattle kept in selected wards of Serengeti and Tarime districts in Mara region. Adult ticks were identified and counted from half body parts of 360 animals which were extensively managed in communal land with natural pastures. Concurrently, blood samples were collected and thereafter DNA extracted and a nested polymerase chain reaction (nPCR) was done using primers specific for p104 gene to detect the presence of T. parva DNA. Ticks were identified into four groups: Amblyomma genus, Boophilus sub-genus of Rhipicephalus genus, other species of Rhipicephalus, and Hyalomma genus. Rhipicephalus genus accounted for 71.8 % of the total ticks, whereas Amblyomma, Boophilus sub-genus of Rhipicephalus genus and Hyalomma constituted 14.1, 14.0 and 0.1 %, respectively. There were more animals (p < 0.05) infested with ticks in Tarime district (96.1 %) than in Serengeti (61.7 %). The average counts of ticks were higher in adult animals (p < 0.05) than in young animals. The overall prevalence of T. parva was 27.7 % and was higher (p < 0.05) in Serengeti (38.3 %) than in Tarime district (16.7 %). However, all animals tested positive for T. parva did not show any clinical signs of East Coast fever (ECF), suggesting the existence of subclinical infection in Tarime zebu. These results suggest that Tarime cattle can tolerate ECF infection and are likely to serve as potential carriers of T. parva to other less-tolerant cattle breeds in mixed herds. Since Tarime cattle are preferred by most farmers with mixed herds, routine screening for T. parva is highly recommended to minimize introduction of infected cattle into an immunologically naive population.

  6. Formula to evaluate efficacy of vaccines and systemic substances against three-host ticks.

    PubMed

    Aguirre, André de Abreu Rangel; Garcia, Marcos Valério; Szabó, Matias Pablo Juan; Barros, Jacqueline Cavalcante; Andreotti, Renato

    2015-05-01

    The control of ticks with vaccines is of global interest. Experimental vaccines incorporate new technologies as soon as they are available. Historically, the main vaccine studies have focused on the one-host cattle tick Rhipicephalus microplus, and efficacy evaluations have been standardised for this tick species. On the other hand, evaluations of vaccine candidates for three-host ticks are being done somewhat arbitrarily and thus comparisons within the current literature on the efficacy of vaccines, as well as other methods of control, are difficult. We herein provide a formula for the evaluation of efficacy of a vaccine designed against three-host ticks that incorporates the whole life cycle of the tick.

  7. Occurrence of Ticks in Cattle in the New Pastoral Farming Areas in Rufiji District, Tanzania.

    PubMed

    Mamiro, Kamilius A; Magwisha, Henry B; Rukambile, Elpidius J; Ruheta, Martin R; Kimboka, Expery J; Malulu, Deusdedit J; Malele, Imna I

    2016-01-01

    Ticks and tick-borne diseases plus trypanosomosis are a constraint to cattle rearing in Tanzania. Rufiji district was not known for important ticks infesting cattle because inhabitants were not engaged in keeping livestock. Not only has settlement of pastoralists and cattle in Rufiji increased the number of cattle but also cattle have been the source of bringing in and spreading of ticks. This study investigated tick species that have been introduced and managed to establish themselves in the new livestock farming areas in cattle in Rufiji. Tick distribution study was undertaken in three villages of Chumbi ward seasonally in 2009, 2011, and 2012. The identified ticks were Amblyomma variegatum (56.10%), Rhipicephalus evertsi (10.25%), R. microplus (27.40%), and R. appendiculatus (6.19%) out of 12940 ticks. Results indicate that ticks are present in the new livestock settlement areas. The occurrence of ticks is correlated with the recent settlement of cattle in the district.

  8. Multiple Pathogens Including Potential New Species in Tick Vectors in Côte d’Ivoire

    PubMed Central

    Ehounoud, Cyrille Bilé; Yao, Kouassi Patrick; Dahmani, Mustapha; Achi, Yaba Louise; Amanzougaghene, Nadia; Kacou N’Douba, Adèle; N’Guessan, Jean David; Raoult, Didier; Fenollar, Florence; Mediannikov, Oleg

    2016-01-01

    Background Our study aimed to assess the presence of different pathogens in ticks collected in two regions in Côte d’Ivoire. Methodology/Principal Findings Real-time PCR and standard PCR assays coupled to sequencing were used. Three hundred and seventy eight (378) ticks (170 Amblyomma variegatum, 161 Rhipicepalus microplus, 3 Rhipicephalus senegalensis, 27 Hyalomma truncatum, 16 Hyalomma marginatum rufipes, and 1 Hyalomma impressum) were identified and analyzed. We identified as pathogenic bacteria, Rickettsia africae in Am. variegatum (90%), Rh. microplus (10%) and Hyalomma spp. (9%), Rickettsia aeschlimannii in Hyalomma spp. (23%), Rickettsia massiliae in Rh. senegalensis (33%) as well as Coxiella burnetii in 0.2%, Borrelia sp. in 0.2%, Anaplasma centrale in 0.2%, Anaplasma marginale in 0.5%, and Ehrlichia ruminantium in 0.5% of all ticks. Potential new species of Borrelia, Anaplasma, and Wolbachia were detected. Candidatus Borrelia africana and Candidatus Borrelia ivorensis (detected in three ticks) are phylogenetically distant from both the relapsing fever group and Lyme disease group borreliae; both were detected in Am. variegatum. Four new genotypes of bacteria from the Anaplasmataceae family were identified, namely Candidatus Anaplasma ivorensis (detected in three ticks), Candidatus Ehrlichia urmitei (in nine ticks), Candidatus Ehrlichia rustica (in four ticks), and Candidatus Wolbachia ivorensis (in one tick). Conclusions/Significance For the first time, we demonstrate the presence of different pathogens such as R. aeschlimannii, C. burnetii, Borrelia sp., A. centrale, A. marginale, and E. ruminantium in ticks in Côte d’Ivoire as well as potential new species of unknown pathogenicity. PMID:26771308

  9. Efficacy of the Bm86 antigen against immature instars and adults of the dog tick Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae).

    PubMed

    Perez-Perez, D; Bechara, G H; Machado, R Z; Andrade, G M; Del Vecchio, R E M; Pedroso, M S; Hernández, M V; Farnós, O

    2010-02-10

    The Bm86 antigen has been used to control ticks of the Boophilus genera in integrated programs that also include the use of acaricides. Because of recent phylogenetic studies have lead to the inclusion of all Boophilus species within the Rhipicephalus genera, we aimed to investigate the efficacy of the Bm86 antigen on the biotic potential of Rhipicephalus sanguineus. Domestic dogs were vaccinated with Bm86 and challenged with the three instars of R. sanguineus. Male and female mongrel dogs were divided into two groups of four animals each, comprising non-vaccinated and vaccinated animals. Immunized dogs were given two doses of an experimental formulation containing 50mug of recombinant Bm86, at 21 days interval while the other group was given placebo, consisting of the same preparation without Bm86. Each dog was challenged 21 days after the last dose with 250 larvae, 100 nymphs and 55 adults (25 females and 30 males) released inside feeding chambers (one per instar) glued to their shaved flank. The effect of the vaccination was evaluated by determining biological parameters of ticks including the yield rates of larvae, nymphs and adult females. Adult females engorged weight, egg mass weight, efficiency rate of conversion to eggs (ERCE) and hatchability. In addition, sera were collected from dogs at 0, 21, 36, 45 and 75 days after the vaccination and used for the detection of specific antibodies by ELISA. Collection rates of larvae, nymphs and adult females fed on vaccinated dogs were significantly (p<0.05) reduced by 38%, 29% and 31%, respectively, as compared with non-vaccinated controls. Significant reductions were also observed in weight of engorged females and egg mass, in ERCE, but not in the hatch rate of ticks fed on immunized dogs. ELISA data revealed a marked and significant increase in optical densities of sera from vaccinated animals after the second dose of Bm86. We concluded that the Bm86 antigen used as a vaccine for dogs reduced the viability and

  10. Identification of Ctenocephalides felis fleas as a host of Rickettsia felis, the agent of a spotted fever rickettsiosis in Yucatań, Mexico.

    PubMed

    Zavala-Velázquez, J E; Zavala-Castro, J E; Vado-Solís, I; Ruiz-Sosa, J A; Moron, C G; Bouyer, D H; Walker, D H

    2002-01-01

    In search for the vector of the recently recognized spotted fever rickettsiosis of the Yucatán, ticks, fleas, and lice were collected from vegetation and dogs in localities where seropositive persons had been found. The arthropods were examined by polymerase chain reaction (PCR) using primers for the genus-specific 17-kDa protein gene followed by restriction fragment length polymorphism (RFLP) and DNA sequencing. Eleven (20%) of 54 pools of Ctenocephalides felis fleas contained DNA of Rickettsia felis. None of 219 Amblyomma cajennense, 474 Rhiphicephalus sanguineus, 258 Boophilus sp. ticks, and 33 Poliplax species lice contained DNA of Rickettsia. The identity of the rickettsial DNA was confirmed as R. felis by PCR/RFLP for the citrate synthase and outer membrane protein A genes and by DNA sequencing. The results indicate that the host of R. felis in Yucatán is C. felis and suggest that the spotted fever rickettsiosis that has infected >5% of the population of the Yucatán and can present as a dengue-like illness is likely to be caused by R. felis.

  11. Arthropod parasites of Nubian ibexes (Capra ibex nubiana) and gazelles (Gazella gazella) in Israel.

    PubMed

    Yeruham, I; Rosen, S; Hadani, A; Braverman, Y

    1999-06-15

    In a 20-year survey the following ectoparasites were collected from Nubian ibexes: larvae of an unidentified Oestrus sp. collected from the nasal cavities, sinuses and horns, hippoboscid flies (Lipoptena chalcomelaena) specific to the Nubian ibex, blood sucking lice (Linognathus africanus) and unidentified biting lice (Damalinia sp.). Ibexes were severely infested with the cattle tick, Boophilus annulatus; a few Hyalomma anatolicum excavatum ticks were also collected. In five ibexes kept in two zoos, showing otitis, Psoroptes cuniculi, was identified, and from skin scrpaings of nine animals with severe dermatitis in three other zoos, Sarcoptes scabiei was isolated. Infestation of Nubian ibexes with sucking and biting lice as well as H. anatolicum excavatum is reported for the first time. The ectoparasites collected from gazelles were: hippoboscid flies (Lipoptena capreoli), calliphorid flies (Lucilia sericata and Calliphora sp.), sucking lice (Linognathus africanus and Solenopotes capillatus) and unidentified biting lice (Damalinia sp.), fleas (Ctenocephalides felis felis), and ticks, B. annulatus, Rhipicepahlus bursa, Rhipicephalus turanicus, H. anatolicum excavatum and H. marginatum rufipes. In skin scrapings of four gazelles with local dermatitis in the fetlocks Chorioptes bovis was identified. Neoschoengastia sp. was found in craters between the claws in three gazelles.

  12. Occurrence and Genotyping of Coxiella burnetii in Ixodid Ticks in Oromia, Ethiopia

    PubMed Central

    Kumsa, Bersissa; Socolovschi, Cristina; Almeras, Lionel; Raoult, Didier; Parola, Philippe

    2015-01-01

    This study was conducted from September 2011 to March 2014 to address the occurrence and genotypes of Coxiella burnetii using molecular methods in ticks collected from domestic animals in Ethiopia. Ticks were tested for C. burnetii by quantitative real-time polymerase chain reaction (qPCR) targeting two different genes followed by multispacer sequence typing (MST). An overall prevalence of 6.4% (54/842) of C. burnetii was recorded. C. burnetii was detected in 28.6% (14/49) of Amblyomma gemma, 25% (31/124) of Rhipicephalus pulchellus, 7.1% (1/14) of Hyalomma marginatum rufipes, 3.2% (2/62) of Am. variegatum, 3.1% (4/128) of Am. cohaerens, 1.6% (1/63) of Rh. praetextatus, and 0.6% (1/153) of Rhipicephalus (Boophilus) decoloratus. Significantly higher overall frequencies of C. burnetii DNA were observed in Am. gemma and Rh. pulchellus than in other tick species (Mantel–Haenszel [MH], P < 0.0001). The overall frequency of C. burnetii was significantly higher (MH, P < 0.0001) in ticks from southeastern districts (Arero, Moyale, and Yabelo) than that from other districts. This study demonstrated the presence of C. burnetii genotype MST 18 in ticks in southeastern districts and genotype MST 20 in ticks in central districts. This study highlights the importance of ticks in the epidemiology of C. burnetii in Ethiopia. PMID:26392155

  13. Infestation and Identification of Ixodid Tick in Cattle: The Case of Arbegona District, Southern Ethiopia

    PubMed Central

    Tamerat, Nateneal; Tuluka, Temesgen

    2016-01-01

    The study was conducted from October 2014 to June 2015 to estimate tick prevalence and identify major tick genera infesting cattle and the associated risk factors in Arbegona district, southern Ethiopia. A total of 2024 adult ticks were collected from main body parts of animals and eight species of ticks which belong to three genera were identified. Questionnaire survey was employed concerning the general case on the tick infestation problems on the cattle. From 384 cattle examined, 291 (75.7%) were found to be infested with one or more types of tick species. The relative prevalence of each genera was Amblyomma (34.9%), Rhipicephalus (Boophilus) (26.6%), Hyalomma (19.2%), and Rhipicephalus (19%). The prevalence of tick infestation in good (65.5%), medium (74%), and poor body condition animal (100%) was found to be statistically significant (p < 0.05). There was also significantly (p < 0.05) higher prevalence in old (98.4%) than adult (78.8%) and young (59.8%) age groups of animals. In the survey, 87.5% of respondents believe that there was tick infestation problem in their locality. This study showed there was high burden and prevalence of ticks that still play major roles in reducing productivity and cause health problems of cattle in the area which call for urgent attention. PMID:28105466

  14. Detection of Babesia spp. in free-ranging Pukus, Kobus vardonii, on a game ranch in Zambia.

    PubMed

    Munang'andu, Hetron Mweemba; Munyeme, Musso; Nambota, Andrew Mubila; Nalubamba, King Shimumbo; Siamudaala, Victor M

    2011-12-01

    Babesia spp. were detected from 4 asymptomatic pukus captured on a game ranch in central Zambia in October 2008. Blood smears were examined in 4 species of aymptomatic free-ranging antelopes, namely the puku (Kobus vordanii), reedbuck (Redunca arundinum), bushbuck (Tragelaphus sylvaticus), and kudu (Tragelaphus strepsiceros), and showed the presence of Babesia parasites only in the puku. In the puku, the prevalence of babesiosis was estimated at 33.3% (n = 12), while the overall prevalence in all examined animals was 8.5% (n = 47). The parasites showed morphological characteristics of paired ring-like stages with the length varying between 1.61 µm and 3.02 µm (mean = 2.12 µm, n = 27; SD = 0.76 µm). Both the infected and non-infected pukus showed good body condition scores (BCS), while the dominant tick species detected from all animals were Rhipicephalus appendiculatus, Rhipicephalus spp., and Boophilus spp. To our knowledge this is the first report of Babesia spp. infection in pukus in Zambia. These findings suggest that wildlife could play an important role in the epidemiology of babesiosis in Zambia.

  15. Species diversity and geographic distribution of hard ticks (Acari: Ixodoidea: Ixodidae) infesting domestic ruminants, in Qazvin Province, Iran.

    PubMed

    Shemshad, Khadijeh; Rafinejad, Javad; Kamali, Karim; Piazak, Norayer; Sedaghat, Mohammad Mahdi; Shemshad, Masoomeh; Biglarian, Akbar; Nourolahi, Fathollah; Valad Beigi, Enshallah; Enayati, Ahmad Ali

    2012-01-01

    This report presents the results of the first faunistic study of hard ticks in Qazvin province of Iran. The primary objective was to determine the species diversity and geographic distribution of hard ticks that parasitize domestic ruminants. Information about the abiotic preferences of these species has been provided. A total of 286 cattle, 1,053 goats, and 2,050 sheep were examined in 13 villages in 28 flocks distributed throughout the studied areas. Total direct body collections of ticks were made from each domestic ruminant. A total of 228 Ixodid specimens belonging to nine species in three different genera were recorded in the areas, including Boophilus annulatus (Say, 1821), Hyalomma anatolicum Koch, 1844, Hyalomma asiaticum (Schulze and Schlettke, 1929), Hyalomma detritum Schulze, 1919, Hyalomma dromedarii Koch, 1844, Hyalomma marginatum Koch 1844, Hyalomma schulzei Olenev, 1931, Rhipicephalus bursa Canestrini and Fanz, 1878 and Rhipicephalus sanguineus (Latreille, 1806). The most abundant species on sheep was R. sanguineus (46.92%), while B. annulatus (6.6%) found only on cattle. A finding of great significance was that R. sanguineus, the main vector of babesiosis, is firmly established throughout the counties. A further objective of the study was to compare the abundance of the major tick species on domestic ruminants. This was carried out at 19 sampling sites. The highest number of ticks was collected in July-August during the hot season.

  16. Relationship between the Prevalence of Ectoparasites and Associated Risk Factors in Free-Range Pigs in Kenya.

    PubMed

    Kagira, John Maina; Kanyari, Paul Njuki; Maingi, Ndicho; Githigia, Samuel Maina; Ng'ang'a, Chege; Gachohi, John

    2013-01-01

    A cross-sectional study was undertaken to determine the prevalence of ectoparasites and possible risk factors in free-range pigs from 135 farms of Busia District, Kenya. Three hundred and six pigs were examined for presence of external parasites using standard parasitological methods. Data on management practices including housing and history of acaricide spraying were also collected. The ectoparasites found in the pigs were Haematopinus suis (96.1%), Sarcoptes scabiei (63.7%), and ticks (29.7%). The tick species included Rhipicephalus appendiculatus (70%), Boophilus decoloratus (31%), and Amblyomma variegatum (12%). The occurrence of the infestations was associated with age, being highest in sows (S. scabiei) and finishers (ticks and H. suis). Male pigs had highest prevalences of H. suis and ticks, while female pigs had highest prevalence of S. scabiei. The prevalence of the parasitic infestations was significantly (P < 0.05) associated with their origin being either lower (H. suis and S. scabiei) or higher (ticks) in pigs originating from divisions with high rainfall. Housed pigs had significantly (P < 0.05) lower prevalence of H. suis and ticks than those from households without pig housing. It is concluded that the free-range pigs have high prevalence of ectoparasites, and effective control strategies focussing on improved animal husbandry and acaricide use should be implemented.

  17. Ticks (Acari: Ixodidae) associated with wildlife and vegetation of Haller park along the Kenyan coastline.

    PubMed

    Wanzala, W; Okanga, S

    2006-09-01

    This artcile describes the results obtained from a tick survey conducted in Haller park along the Kenyan coastline. The survey aimed at evaluating tick-host associations, assessing tick population density, and providing baseline information for planning future tick control and management in the park. Ticks (2,968) were collected by handpicking from eight species of wildlife and by dragging in 14 selected sites within the park. A considerable proportion of ticks were also collected from leaves, stems, and bark of most dominant trees, namely, Casuarina equisetifolia L. (Forst. and Forst.), Cocos nucifera L., Adansonia digitata L., Musa paradisiaca L., and Azadiracta indica Adr. Juss. Dragging was conducted in sites predominantly occupied by Cynodon dactylon L. (Pers.), Cenchrus ciliaris L., Stenotaphrum dimidiatum L. (Kuntze.) Brongn., and Brachiaria xantholeuca Hack. Ex Schinz Stapf. and Loudetia kagerensis K. Schum. Hutch. Eight tick species were identified, and the collection included Rhipicephalus pravus Dönitz 1910, Rhipicephalus pulchellus Gerstäcker 1873, Hyalomma marginatum rufipes Koch 1844, Amblyomma gemma Dönitz 1910, Amblyomma hebraeum Koch 1844, Amblyomma sparsum Neumann 1899, Amblyomma nuttalli Dönitz 1909, and Boophilus decoloratus Koch 1844. Given that the identified tick species are known to parasitize humans as well as livestock, there exist risks of emergence of zoonotic infections mediated by tick vectors. In the recreational environment of Haller park, where tick vectors share habitats with hosts, there is a need to develop sustainable and effective tick control and management strategies to minimize economic losses that tick infestation may cause.

  18. Ticks (Acari: Ixodidae) infesting cattle and African buffaloes in the Tsavo conservation area, Kenya.

    PubMed

    Kariuki, Edward K; Penzhorn, Barend L; Horak, Ivan G

    2012-11-21

    Several ixodid tick species are shared between domestic cattle and African buffaloes (Syncerus caffer). So too, are a number of tick-borne diseases. The aim of the study was to compare the species composition of ticks that infest cattle and buffaloes utilising the same habitat within the Tsavo Conservation Area, Kenya. To this end, 25 cattle and 62 buffaloes were each opportunistically sampled for ticks on a single occasion in February 2010. Eight species, namely Amblyomma gemma, Amblyomma lepidum, Hyalomma albiparmatum, Hyalomma rufipes, Hyalomma truncatum, Rhipicephalus evertsi evertsi, Rhipicephalus pravus and Rhipicephalus pulchellus infested both cattle and buffaloes. Three species, Rhipicephalus (Boophilus) sp., Rhipicephalus kochi, and Rhipicephalus muehlensi were collected only from cattle, and three species, Hyalomma impeltatum, Rhipicephalus humeralis and Rhipicephalus praetextatus were present only on buffaloes. The attachment sites of the various tick species were also recorded. New locality records for H. impeltatum and H. truncatum and the first confirmed locality record for Rhipicephalus praetextatus sensu stricto in Kenya were documented.

  19. Tick loads in cattle raised on sweet and sour rangelands in the low-input farming areas of South Africa.

    PubMed

    Marufu, Munyaradzi C; Chimonyo, Michael; Mapiye, Cletos; Dzama, Kennedy

    2011-02-01

    The objective of this study was to compare tick loads and prevalence in Nguni and non-descript cattle in the sweet (palatable throughout the year) and sour (palatable only in the rainy season) communal rangelands of the Eastern Cape Province, South Africa. Engorged adult female ixodid ticks were collected and identified seasonally from 144 cattle raised on sweet and sour rangelands from August 2007 to April 2008. Three tick species were identified in the sweet and sour rangelands namely Rhipicephalus appendiculatus, Rhipicephalus (Boophilus) decoloratus, and Rhipicephalus evertsi evertsi with prevalences of 71.1%, 29.2%, and 40.2%, respectively. Hyalomma species (19.0%) occurred only in the sour rangeland. Higher tick counts were recorded in the hot-wet season than in the cool-dry season (P < 0.05). Cattle in the sweet rangeland had significantly lower tick loads than those in the sour rangeland in all the seasons except the hot-dry season. The Nguni breed had lower (P < 0.05) tick loads of R. appendiculatus in the hot-wet and post-rainy season and Hyalomma species in all seasons than the non-descript cattle. The use of a tick-resistant Nguni breed in the integrated control of ticks on cattle in the communal areas of South Africa is recommended.

  20. Primary report on distribution of tick fauna in Iran.

    PubMed

    Rahbari, Sadegh; Nabian, Sedighe; Shayan, Parviz

    2007-09-01

    A tick survey was carried out in four different geographical areas of Iran, where the majority of the domestic ruminants in Iran exist. About 1,500 sheep, 1,200 goats and 500 cattle of 12 herds in different provinces lying in the corresponding zones were inspected for tick infestation. The occurrence of ticks on cattle, sheep and goats were 62, 55 and 57%, respectively, with no differences between the zones. The mean number of ticks on each animal was low (10-20 ticks per animal). Ixodid ticks were found throughout the year, whereas the soft tick Ornithodoros sp., which occurred in mountainous area with a significant difference in abundance, showed a clear pattern of seasonality, being generally present from November to March. The largest numbers of adult ixodid ticks were generally present from April to August. Rhipicephalus, Haemaphysalis and Dermacentor ticks occurred in the mountainous area, whereas Boophilus and Ixodes ticks were only present in the Caspian region. Hyalomma were very abundant in each zone but especially in the mountainous area, whereas Ixodes ticks were the minor genus.

  1. Bartonella and Rickettsia in arthropods from the Lao PDR and from Borneo, Malaysia.

    PubMed

    Kernif, Tahar; Socolovschi, Cristina; Wells, Konstans; Lakim, Maklarin B; Inthalad, Saythong; Slesak, Günther; Boudebouch, Najma; Beaucournu, Jean-Claude; Newton, Paul N; Raoult, Didier; Parola, Philippe

    2012-01-01

    Rickettsioses and bartonelloses are arthropod-borne diseases of mammals with widespread geographical distributions. Yet their occurrence in specific regions, their association with different vectors and hosts and the infection rate of arthropod-vectors with these agents remain poorly studied in South-east Asia. We conducted entomological field surveys in the Lao PDR (Laos) and Borneo, Malaysia by surveying fleas, ticks, and lice from domestic dogs and collected additional samples from domestic cows and pigs in Laos. Rickettsia felis was detected by real-time PCR with similar overall flea infection rate in Laos (76.6%, 69/90) and Borneo (74.4%, 268/360). Both of the encountered flea vectors Ctenocephalides orientis and Ctenocephalides felis felis were infected with R. felis. The degrees of similarity of partial gltA and ompA genes with recognized species indicate the rickettsia detected in two Boophilus spp. ticks collected from a cow in Laos may be a new species. Isolation and further characterization will be necessary to specify it as a new species. Bartonella clarridgeiae was detected in 3/90 (3.3%) and 2/360 (0.6%) of examined fleas from Laos and Borneo, respectively. Two fleas collected in Laos and one flea collected in Borneo were co-infected with both R. felis and B. clarridgeiae. Further investigations are needed in order to isolate these agents and to determine their epidemiology and aetiological role in unknown fever in patients from these areas.

  2. Anaplasma Infection in Ticks, Livestock and Human in Ghaemshahr, Mazandaran Province, Iran

    PubMed Central

    Hosseini-Vasoukolaei, Nasibeh; Oshaghi, Mohammad Ali; Shayan, Parviz; Vatandoost, Hassan; Babamahmoudi, Farhang; Yaghoobi-Ershadi, Mohammad Reza; Telmadarraiy, Zakkyeh; Mohtarami, Fatemeh

    2014-01-01

    Background: Anaplasmosis is an important issue for animal breeders in terms of economic losses as well as a health concern to human. Ticks are considered as the main vector of this disease. Lack of documented information about Anaplasma species in Iran was the scope of this study to determine the population of ticks and the presence of Anaplasma in ticks, domestic ruminants and also human beings in northern Iran. Methods: A total of 101 unengorged hard ticks, 78 domestic ruminants and 40 human blood samples collected from Ghaemshahr, Mazandaran Province, northern Iran were tested by nested PCR against 16s rRNA gene of Anaplasma species. Results: Positive PCR was found in 50 ticks, 28 sheep, 2 cattle, one goat, and 10 human specimens. Sequence analysis of the PCR products confirmed presence of A. ovis in two Rhipicephalus sanguineus and two Ixodes ricinus ticks, one human and 4 sheep samples. Moreover one Boophilus annulatus tick and one sheep sample were infected with A. bovis. Furthermore one sample of sheep was infected with A. centrale. Conclusion: This study is the first report of tick infection to A. ovis, A. bovis and human infection to A. ovis in Iran. The result of this study is a survey of Anaplasma infections from ticks, domestic animals and human in Iran which help to have appropriate prevention measures for anaplasmosis. PMID:26114134

  3. Molecular identification of tick-borne pathogens in Nigerian ticks.

    PubMed

    Ogo, Ndudim Isaac; de Mera, Isabel G Fernández; Galindo, Ruth C; Okubanjo, Oluyinka O; Inuwa, Hauwa Mairo; Agbede, Rowland I S; Torina, Alessandra; Alongi, Angelina; Vicente, Joaquín; Gortázar, Christian; de la Fuente, José

    2012-07-06

    A molecular epidemiology investigation was undertaken in two Nigerian states (Plateau and Nassarawa) to determine the prevalence of pathogens of veterinary and public health importance associated with ticks collected from cattle and dogs using PCR, cloning and sequencing or reverse line blot techniques. A total of 218 tick samples, Amblyomma variegatum (N=153), Rhipicephalus (Boophilus) decoloratus (N=45), and Rhipicephalus sanguineus (N=20) were sampled. Pathogens identified in ticks included piroplasmids (Babesia spp., Babesia bigemina and Babesia divergens), Anaplasma marginale and Rickettsia africae. Piroplasmids were identified in A. variegatum, A. marginale was found in R. decoloratus, while R. africae was detected in all tick species examined. Ehrlichia spp. and Theileria spp. were not identified in any of the ticks examined. Of the 218 ticks examined, 33 (15.1%) contained pathogen DNA, with the presence of B. divergens and R. africae that are zoonotic pathogens of public health and veterinary importance. The variety of tick-borne pathogens identified in this study suggests a risk for the emergence of tick-borne diseases in domestic animals and humans, especially amongst the Fulani pastoralists in Plateau and Nassarawa states of Nigeria.

  4. Biogeography of Tick-Borne Bhanja Virus (Bunyaviridae) in Europe

    PubMed Central

    Hubálek, Zdenek

    2009-01-01

    Bhanja virus (BHAV) is pathogenic for young domestic ruminants and also for humans, causing fever and affections of the central nervous system. This generally neglected arbovirus of the family Bunyaviridae is transmitted by metastriate ticks of the genera Haemaphysalis, Dermacentor, Hyalomma, Rhipicephalus, Boophilus, and Amblyomma. Geographic distribution of BHAV covers southern and Central Asia, Africa, and southern (partially also central) Europe. Comparative biogeographic study of eight known natural foci of BHAV infections in Europe (in Italy, Croatia, Bulgaria, Slovakia) has revealed their common features. (1) submediterranean climatic pattern with dry growing season and wet mild winter (or microlimatically similar conditions, e.g., limestone karst areas in central Europe), (2) xerothermic woodland-grassland ecosystem, with plant alliances Quercetalia pubescentis, Festucetalia valesiacae, and Brometalia erecti, involving pastoral areas, (3) presence of at least one of the tick species Haemaphysalis punctata, Dermacentor marginatus, Rhipicephalus bursa, and/or Hyalomma marginatum, and (4) presence of ≥60% of the 180 BHAV bioindicator (157 plant, 4 ixodid tick, and 19 vertebrate spp.). On that basis, Greece, France (southern, including Corsica), Albania, Spain, Hungary, European Turkey, Ukraine (southern), Switzerland (southern), Austria (southeastern), Germany (southern), Moldova, and European Russia (southern) have been predicted as additional European regions where BHAV might occur. PMID:20182535

  5. Molecular characterization of DSC1 orthologs in invertebrate species.

    PubMed

    Cui, Ying-Jun; Yu, Lin-Lin; Xu, Hai-Jun; Dong, Ke; Zhang, Chuan-Xi

    2012-05-01

    DSC1 and BSC1 are two founding members of a novel family of invertebrate voltage-gated cation channels with close structural and evolutionary relationships to voltage-gated sodium and calcium channels. In this study, we searched the published genome sequences for DSC1 orthologs. DSC1 orthologs were found in all 48 insect species, and in other invertebrate species belonging to phyla Mollusca, Cnidaria, Hemichordata and Echinodermata. However, DSC1 orthologs were not found in four arachnid species, Ixodes scapularis, Rhipicephalus microplus, Tetranychus urticae and Varroa destructor, two species in Annelida or any vertebrate species. We then cloned and sequenced NlSC1 and BmSC1 full-length cDNAs from the brown planthopper (Nilaparvata lugens) and the silkworm (Bombyx mori), respectively. NlSC1 and BmSC1 share about 50% identity with DSC1, and the expression of NlSC1 and BmSC1 transcripts was most abundant in the head and antenna in adults. All DSC1 orthologs contain a unique and conserved DEEA motif, instead of the EEEE or EEDD motif in classical calcium channels or the DEKA motif in sodium channels. Phylogenetic analyses revealed that DSC1 and its orthologs form a separate group distinct from the classical voltage-gated sodium and calcium channels and constitute a unique family of cation channels. The DSC1/BSC1-family channels could be potential targets of new and safe insecticides for pest control.

  6. The ticks (Acari: Ixodida: Argasidae, Ixodidae) of Bolivia.

    PubMed

    Mastropaolo, Mariano; Beltrán-Saavedra, L Fabián; Guglielmone, Alberto A

    2014-03-01

    The tick species reported in Bolivia are reviewed here as (1) endemic or established: Ornithodoros echimys, O. guaporensis, O. hasei, O. kohlsi, O. mimon, O. peropteryx, O. rostratus, Otobius megnini, Amblyomma auricularium, A. cajennense, A. calcaratum, A. coelebs, A. dubitatum, A. humerale, A. incisum, A. longirostre, A. naponense, A. nodosum, A. oblongoguttatum, A. ovale, A. parvitarsum, A. parvum, A. pecarium, A. pseudoconcolor, A. rotundatum, A. scalpturatum, A. tigrinum, A. triste, Dermacentor nitens, Haemaphysalis juxtakochi, H. leporispalustris, I. boliviensis, I. cooleyi, I. luciae, Rhipicephalus microplus, R. sanguineus, and (2) erroneously reported: Ornithodoros puertoricensis, O. talaje, O. turicata, Amblyomma americanum, A. maculatum, A. multipunctum, Ixodes ricinus, I. scapularis, Rhipicephalus annulatus. Many of these records are lacking locality and/or host, and some of them need new findings for confirmation. Some of the species recorded may represent a threat for human and animal health, therefore would be of great value to make a countrywide survey of ticks in order to update the information presented in this work.

  7. Molecular survey of hard ticks in endemic areas of tick-borne diseases in China.

    PubMed

    Lu, Xin; Lin, Xian-Dan; Wang, Jian-Bo; Qin, Xin-Cheng; Tian, Jun-Hua; Guo, Wen-Ping; Fan, Fei-Neng; Shao, Renfu; Xu, Jianguo; Zhang, Yong-Zhen

    2013-06-01

    Over the past several years, there was a substantial increase in the number of cases of known and novel tick-borne infections in humans in China. To better understand the ticks associated with these infections, we collected hard ticks from animals or around livestock shelters in 29 localities in 5 provinces (Beijing, Henan, Hubei, Inner Mongolia, and Zhejiang) where cases of tick-borne illness were reported. We collected 2950 hard ticks representing 7 species of 4 genera (Dermacentor sinicus, Haemaphysalis flava, Haemaphysalis longicornis, Ixodes granulatus, Ixodes persulcatus, Rhipicephalus microplus, and Rhipicephalus sanguineus). These ticks were identified to species using morphological characters initially. We then sequenced the mitochondrial small subunit rRNA (12S rRNA) gene, cytochrome oxidase subunit 1 (COI) gene, and the second internal transcribed spacer (ITS2) gene of these ticks, and conducted phylogenetic analyses. Our analyses showed that the molecular and morphological data are consistent in the identification of the 7 tick species. Furthermore, all these 7 tick species from China were genetically closely related to the same species or related species found outside China. Rapid and accurate identification and long-term monitoring of these ticks will be of significance to the prevention and control of tick-borne diseases in China.

  8. Streptomyces sp. LK3 mediated synthesis of silver nanoparticles and its biomedical application.

    PubMed

    Karthik, L; Kumar, Gaurav; Kirthi, A Vishnu; Rahuman, A A; Bhaskara Rao, K V

    2014-02-01

    In the present study, the marine actinobacteria mediated biosynthesis of silver nanoparticles (AgNps) was achieved using Streptomyces sp LK3. The synthesized AgNps showed the characteristic absorption spectra in UV-vis at 420 nm, which confirmed the presence of nanoparticles. XRD analysis showed intense peaks at 2θ values of 27.51°, 31.87°, 45.57°, 56.56°, 66.26°, and 75.25° corresponding to (210), (113), (124), (240), (226), and (300) Bragg's reflection based on the fcc structure of AgNps. The FTIR spectra exhibited prominent peaks at 3,417 cm(-1) (OH stretching due to alcoholic group) and 1,578 cm(-1) (C=C ring stretching). TEM micrograph showed that the synthesized AgNps were spherical in shape with an average size of 5 nm. Surface morphology and topographical structure of the synthesized AgNps were dignified by AFM. The synthesized AgNps showed significant acaricidal activity against Rhipicephalus microplus and Haemaphysalis bispinosa with LC50 values of 16.10 and 16.45 mg/L, respectively. Our results clearly indicate that AgNps could provide a safer alternative to conventional acaricidal agents in the form of a topical antiparasitic formulation. The present study aimed to develop a novel, cost-effective, eco-friendly actinobacteria mediated synthesis of AgNps and its antiparasitic activity.

  9. Molecular evidence of spotted fever group rickettsiae and Anaplasmataceae from ticks and stray dogs in Bangladesh.

    PubMed

    Qiu, Yongjin; Nakao, Ryo; Thu, May June; Akter, Shirin; Alam, Mohammad Zahangir; Kato, Satomi; Katakura, Ken; Sugimoto, Chihiro

    2016-03-01

    Emerging tick-borne diseases (TBDs) are important foci for human and animal health worldwide. However, these diseases are sometimes over looked, especially in countries with limited resources to perform molecular-based surveys. The aim of this study was to detect and characterize spotted fever group (SFG) rickettsiae and Anaplasmataceae in Bangladesh, which are important tick-borne pathogens for humans and animals worldwide. A total of 50 canine blood samples, 15 ticks collected from dogs, and 154 ticks collected from cattle were screened for the presence of SFG rickettsiae and Anaplasmataceae using molecular-based methods such as PCR and real-time PCR. The sequence analysis of the amplified products detected two different genotypes of SFG rickettsiae in ticks from cattle. The genotype detected in Rhipicephalus microplus was closely related to Rickettsia monacensis, while the genotype detected in Haemaphysalis bispinosa was closely related to Rickettsia sp. found in Korea and Japan. Anaplasma bovis was detected in canine blood and ticks (Rhipicephalus sanguineus and H. bispinosa). Unexpectedly, the partial genome sequence of Wolbachia sp., presumably associated with the nematode Dirofilaria immitis, was identified in canine blood. The present study provides the first molecular evidence of SFG rickettsiae and A. bovis in Bangladesh, indicating the possible emergence of previously unrecognized TBDs in this country.

  10. Ticks infesting wild and domestic animals and humans of Sri Lanka with new host records.

    PubMed

    Liyanaarachchi, D R; Rajakaruna, R S; Dikkumbura, A W; Rajapakse, R P V J

    2015-02-01

    An island-wide collection of tick species infesting humans, domesticated and wild animals and questing ticks in domestic and peridomestic environments was carried out during 2009-2011. A total of 30,461 ticks were collected from 30 different hosts and free living stages from the ground. The collection consisted of 22 tick species from 30 different hosts recording 12 tick species from humans, 19 from domesticated animals and 21 from wild animals, with a total of 97 new host records. The most common tick species on humans were Dermacentor auratus and Amblyomma testudinairum, while Haemaphysalis intermedia, Rhipicephalus microplus and Rhipicephalus sanguineus were common in domesticated and wild animals sharing 20 host species. Among the questing ticks, immature D. auratus was the most abundant. Humans and domesticated animals were mostly infested by the nymphal stages while adult ticks were found on wild animals. High number of new host records could be due to domestic animals picking tick species from wildlife and vise versa at the human/animal interface. Habitat destruction due to forest fragmentation has lead to wild animals roaming in urban and semi-urban neighbourhoods increasing the interactions of wild animals with domesticated animals. Wild animals play a significant role as a reservoir of many tick borne infections which can easily be spread to domesticated animals and then to humans via tick infestations. Data in this paper are useful for those interested in tick infesting wild and domestic animals and humans in describing the zoonotic potential of tick borne infections.

  11. Distribution of Spotted Fever Group Rickettsiae in Hard Ticks (Ixodida: Ixodidae) from Panamanian Urban and Rural Environments (2007-2013).

    PubMed

    Bermúdez, Sergio E; Castro, Angélica M; Trejos, Diomedes; García, Gleydis G; Gabster, Amanda; Miranda, Roberto J; Zaldívar, Yamitzel; Paternina, Luis E

    2016-06-01

    Tick-borne rickettsiosis is an important emerging disease in Panama; to date, there have been 12 confirmed cases, including eight fatalities. To evaluate the distribution of rickettsiae in Panamanian ticks, we collected questing and on-host ticks in urban and rural towns in elevations varying between 0 and 2300 m. A total of 63 sites (13 urban and 50 rural towns) were used to develop models of spatial distributions. We found the following tick species: Rhipicephalus sanguineus s.l. (present in 54 of 63 towns and cities), Amblyomma mixtum (45/63), Dermacentor nitens (40/63), A. ovale (37/63), Rhipicephalus microplus (33/63), A. oblongoguttatum (33/63), Ixodes affinis (3/63), and Ixodes boliviensis (2/63). Rhipicephalus sanguineus s.l. was present in urban and rural towns, and other species were present only in rural towns. DNA was extracted from 408 R. sanguineus s.l., 387 A. mixtum, 103 A. ovale, and 11 A. oblongoguttatum and later tested for rickettsiae genes using PCR. Rickettsia DNA was detected in ticks from 21 of 63 localities. Rickettsia rickettsii was detected in five A. mixtum (1.29%), and Candidatus "Rickettsia amblyommii" was found in 138 A. mixtum (35%), 14 R. sanguineus (3.4%), and one A. ovale (0.9%). These results suggest that much of rural Panama is suitable for the expansion of tick populations and could favor the appearance of new tick-borne rickettsiosis outbreaks.

  12. Ticks of Australia. The species that infest domestic animals and humans.

    PubMed

    Barker, Stephen C; Walker, Alan R

    2014-06-18

    The book Australian Ticks by F.H.S. Roberts (1970) is a land-mark in Australian tick biology. But it is time for a new and improved book on the ticks of Australia. The present book has identification guides and accounts of the biology and diseases associated with the 16 species of ticks that may feed on domestic animals and humans in Australia. These comprise five argasid (soft) ticks: Argas persicus (poultry tick), Argas robertsi (Robert's bird tick), Ornithodoros capensis (seabird soft tick), O. gurneyi (kangaroo soft tick), Otobius megnini (spinose ear tick); and 11 ixodid (hard) ticks, Amblyomma triguttatum (ornate kangaroo tick), Bothriocroton auruginans (wombat tick), B. hydrosauri (southern reptile tick), Haemaphysalis bancrofti (wallaby tick), H. longicornis (bush tick), Ixodes cornuatus (southern paralysis tick), I. hirsti (Hirst's marsupial tick), I. holocyclus (paralysis tick), I. tasmani (common marsupial tick), Rhipicephalus (Boophilus) australis (Australian cattle tick) and R. sanguineus (brown dog tick).  We use an image-matching system to identify ticks, much like the image-matching systems used in field-guides for birds and flowers. Ticks may be identified by drawings that emphasise unique matrices of uniformly defined morphological characters that, together, allow these 16 ticks to be identified by morphology unequivocally. The species accounts have seven sections: (i) General; (ii)  Differential diagnosis; (iii) Hosts; (iv) Life-cycle and seasonality; (v) Disease; (vi) Habitat and geographic distribution; (vii) Genes and genomes; and (viii) Other information. There are 71 figures and tables, including a glossary character matrices, drawings of life-cycles, drawings of genera, species, and colour photographs of tick biology.

  13. Immunization of Cattle with Tick Salivary Gland Extracts

    PubMed Central

    Nikpay, Ali; Nabian, Sedigheh

    2016-01-01

    Background: Rhipicephalus (Boophilus) annulatus tick is one of the most important ectoparasite of cattle. Recently, several laboratories in the world have been concentrated on immunizing cattle against tick using various types of tissue extracts of ticks. The aim of this study was to evaluate the effect of immunization of cattle with tick salivary gland extract on biological parameters of ticks and humoral immune responses of cattle. Methods: Fourteen more dominant protein bands identified as immunogenic by Western-blot analysis were eluted from polyacrylamide gel. Test and control groups were injected three times with eluted proteins and sterile PBS (pH= 7.2) respectively with equivalent amount of adjuvant. After four weeks a tick challenge was performed. Finally, biological parameters of collected engorged female ticks were recorded and humoral immune responses to immunization measured by ELISA. Results: The results indicated immunization of cattle resulted in reduction in mean tick counts, attachment, engorgement weights, feeding index, egg mass weight, hatchability and fertility index (respectively 63.1%, 62.6%, 30.2%, 36.4%, 40%, 78.7% and 13.3%) and increased duration of feeding, preoviposition and incubation period of eggs (respectively 8.6%, 45 and 31.34%). All changes were statistically significant (P< 0.05). Results showed an increase in antibody production of test group from the first week after immunization. The antibody level was boosted following tick infestation. Conclusion: This investigation indicates that immunization of cattle with these antigens could induce a protective immune response against Rh. (B.) annulatus tick that would be expected to provide a safe non-chemical means of tick control. PMID:27308287

  14. Protozoan and bacterial pathogens in tick salivary glands in wild and domestic animal environments in South Africa.

    PubMed

    Berggoetz, M; Schmid, M; Ston, D; Wyss, V; Chevillon, C; Pretorius, A-M; Gern, L

    2014-03-01

    A total of 7364 ticks belonging to 13 species was collected from 64 game animals (belonging to 11 species) and from 64 livestock animals (cattle and sheep) living in close vicinity at 6 localities in 3 South African Provinces (Free State, Mpumalanga, and Limpopo). The geographic distribution of all tick species was congruent with the literature except for Haemaphysalis silacea. From each infested host, a maximum of 10 males and 10 females of each tick species were dissected to isolate the salivary glands. Salivary glands were screened for tick-borne pathogens using polymerase chain reaction followed by reverse line blotting and sequencing. This approach allowed us to evaluate the exposure of wild and domestic hosts to tick-borne pathogens in their respective environments. Among the 2117 examined ticks, 329 (15.5%), belonging to 8 species, were infected and harboured 397 infections. Among those, 57.7% were identified to species level and were assigned to 23 pathogen species of the genera Babesia, Theileria, Anaplasma, and Ehrlichia. In 3 out of 6 localities, salivary glands from ticks infesting wild ruminants displayed significantly higher infection prevalence and pathogen mean density than salivary glands from ticks infesting livestock animals. Four piroplasm species [Theileria bicornis, Babesia sp. (sable), Theileria sp. (giraffe), and Theileria sp. (kudu)] were detected for the first time in ticks. The tick species Rhipicephalus evertsi evertsi, Rhipicephalus (Boophilus) decoloratus, Hyalomma rufipes, Rhipicephalus appendiculatus, and Amblyomma hebraeum were associated with a broader pathogen range than previously known, and thus new vector-pathogen combinations are described. In addition, previously unknown coinfection patterns in tick salivary glands are reported.

  15. Smallholder pig production: prevalence and risk factors of ectoparasites.

    PubMed

    Braae, U C; Ngowi, H A; Johansen, M V

    2013-09-01

    A cross-sectional study was carried out in the Mbeya Region, Tanzania, with the aim of describing the distribution and diversity of ectoparasites on pigs, within confinement and free-range production systems of smallholder farms. A total of 128 farms were surveyed, with 96 practising confinement and 32 practising free-range production systems. The prevalence of ectoparasites on pigs within confinement and free-range production systems was 24% and 84%, respectively. Logistic regression analyses revealed that keeping pigs in a free-range system and the presence of neighbouring pigs were risk factors for ectoparasites. Within the confinement system, contact with neighbouring pigs and the time interval (in months) since last ectoparasitic treatment were additionally identified as risk factors. The prevalence of Haematopinus suis was 20% in confined pigs and 63% among free-range pigs. Free-ranging of pigs and presence of neighbouring pigs were also identified as risk factors for the presence of lice. Three species of fleas were identified; Tunga penetrans, Echidnophaga gallinacea and Ctenocephalides canis. The prevalence of fleas was 5% and 13% within confined and free-range, respectively. Two pigs (2%) were found infested with Sarcoptes scabiei var. suis. Ticks found belonged to four genera; Amblyomma spp., Rhipicephalus spp., Haemaphysalis spp., and Boophilus spp. The prevalence of hard ticks among the free-range pigs was 50%. Ectoparasites were more prevalent in the free-range system although highly prevalent within both production systems. Keeping pigs in a free-range system and contact with neighbouring pigs were main risk factors for the presence of ectoparasites. Confinement was highly effective as a preventive tool against hard ticks.

  16. Parasites of domestic and wild animals in South Africa. XV. The seasonal prevalence of ectoparasites on impala and cattle in the Northern Transvaal.

    PubMed

    Horak, I G

    1982-06-01

    The prevalence of ectoparasites on a total of 36 impala (Aepyceros melampus) slaughtered monthly from February 1975 to February 1976 and a total of 24 cattle slaughtered monthly from March 1976 to March 1977 in the Nylsvley Provincial Nature Reserve was determined. Six species of ixodid ticks were collected from the impala and these, in order of abundance, were: Rhipicephalus evertsi evertsi, Rhipicephalus appe. diculatus, Amblyomma hebraeum, Boophilus decoloratus, Ixodes cavipalpus and Hyalomma marginatum rufipes. Only 340 (2,7%) of the 12 757 ticks collected from the impala were adult. The 4 species of lice present on the impala were, in order of abundance: Damalinia aepycerus, Linognathus aepycerus, Damalinia elongata and Linognathus nevilli. The cattle harboured 8 species of ixodid ticks. In order of abundance, these were: R. appendiculatus, R. evertsi evertsi, A. hebraeum, Hyalomma truncatum, H. marginatum rufipes, B. decoloratus, Rhipicephalus simus and I. cavipalpus. A total of 14 186 ticks was collected from the cattle and of these 4 660 (32,9%) were adults. Clear seasonal prevalences could be determined for certain ticks only. Adult A. hebraeum reached peak numbers on cattle from November to March, adult H. marginatum rufipes from December to February and adult H. truncatum during January and February. Larvae of R. appendiculatus reached peak numbers on cattle and impala from March or April to July, nymphae from June to October and adults from December to March. Peak numbers of larvae of R. evertsi evertsi were recovered from impala from May to July and nymphae during July, while adults were present on cattle throughout the survey period, with peaks being recorded during December and February.

  17. Grooming in impala: role of oral grooming in removal of ticks and effects of ticks in increasing grooming rate.

    PubMed

    Mooring, M S; McKenzie, A A; Hart, B L

    1996-01-01

    In Experiment 1, five adult female impala were fitted with harnesses that restrained oral self-grooming of the anterior part of the body. At the same time, six cohoused female impala were fitted with control harnesses that allowed normal oral grooming. The impala were allowed to habituate to the harnesses for 10 days, and both groups were then exposed to larval ticks (Boophilus decoloratus) by herding them into a tick-seeding corral. During the third week following tick seeding, when female ticks were estimated to have developed into engorging adults, the impala were immobilized, tick numbers on the animals sampled by patch sampling, and the harnesses removed. Observations continued for 5 days following removal of the harnesses. Twenty-minute focal observations were conducted daily on each impala during the habituation, tick-seeded, and postharness phases. Restrained impala had a median of 20 times more adult female ticks (both engorged and unengorged) than control impala. Oral grooming, which had been suppressed in the restrained impala during habituation and tick-seeded phases, increased 10-fold once the harnesses were removed and occurred 2.5 times more frequently than in control impala during the postharness phase. In Experiment 2, 15 adult female impala were seeded with larval ticks as in Experiment 1; in week 3 after tick seeding all ticks were removed from animals by application of an acaricide. Grooming was recorded during 3 weeks of baseline observations prior to tick seeding, 3 weeks after tick seeding, and then for 3 weeks beginning 1 week after acaricide treatment. Oral grooming and scratch grooming significantly increased from baseline during tick seeding and significantly declined following removal of the ticks with acaracide. Taken together, the two experiments demonstrate that oral grooming is very effective and important in removing fitness-compromising ticks in free-ranging impala. Correspondingly, exposure to, and subsequent infestation by, ticks

  18. Vectors of Crimean Congo Hemorrhagic Fever Virus in Iran

    PubMed Central

    Telmadarraiy, Zakkyeh; Chinikar, Sadegh; Vatandoost, Hassan; Faghihi, Faezeh; Hosseini-Chegeni, Asadollah

    2015-01-01

    Background: Ticks are important vectors and reservoirs of Crimean Congo Hemorrhagic Fever (CCHF) virus. Human beings may be infected whenever the normal life cycle of the infected ticks on non-human vertebrate hosts is interrupted by the undesirable presence of humans in the cycle. A total of 26 species of Argasid and Ixodid ticks have been recorded in Iran; including nine Hyalomma, two Rhipicephalus, two Dermacentor, five Haemaphysalis, two Boophilus, one Ixodes and two Argas as well as three Ornithodoros species as blood sucking ectoparasites of livestock and poultries. The present paper reviews tick vectors of CCHF virus in Iran, focusing on the role of ticks in different provinces of Iran using reverse transcription polymerase chain reaction (RT-PCR) assay. Methods: During ten years study, 1054 tick specimens; including two species of Argasidae and 17 species of Ixodidae were examined for their infection to CCHF virus genome. The output of all studies as well as related publications were discussed in the current paper. Results: The results show that Rhipicephalus sanguineus, Hyalomma marginatum, H. anatolicum, H. asiaticum and H. dromedarii were known as the most frequent species which were positive for CCHF virus. Conclusion: The status of ticks which were positive for CCHF virus revealed that unlike the most common idea that Hyalomma species are the most important vectors of CCHF virus, other ticks including Rhipicephalus, Haemaphysalis and Dermacentor can be reservoir of this virus; thus, considering geographical distribution, type of host and environmental conditions, different tick control measurements should be carried out in areas with high incidence of CCHF disease. PMID:26623426

  19. Detection of Theileria and Babesia in brown brocket deer (Mazama gouazoubira) and marsh deer (Blastocerus dichotomus) in the State of Minas Gerais, Brazil.

    PubMed

    da Silveira, Júlia A G; Rabelo, Elida M L; Ribeiro, Múcio F B

    2011-04-19

    Intraerythrocytic protozoan species of the genera Theileria and Babesia are known to infect both wild and domestic animals, and both are transmitted by hard-ticks of the family Ixodidae. The prevalences of hemoprotozoa and ectoparasites in 15 free-living Mazama gouazoubira, two captive M. gouazoubira and four captive Blastocerus dichotomus from the State of Minas Gerais, Brazil, have been determined through the examination of blood smears and the use of nested polymerase chain reaction (nPCR). The cervid population was inspected for the presence of ticks and any specimens encountered were identified alive under the stereomicroscope. Blood samples were collected from all 21 animals, following which blood smears were prepared, subjected to quick Romanowsky staining and examined under the optical microscope. DNA was extracted with the aid of commercial kits from cervid blood samples and from tick salivary glands. The nPCR assay comprised two amplification reactions: the first was conducted using primers specific for a 1700 bp segment of the 18S rRNA gene of Babesia and Theileria species, whilst the second employed primers designed to amplify a common 420 bp Babesia 18S rRNA fragment identified by aligning sequences from Babesia spp. available at GenBank. The ticks Amblyomma cajennense, Rhipicephalus microplus and Dermacentor nitens were identified in various of the cervids examined. Of the animals investigated, 71.4% (15/21) were infected with hemoprotozoa, including Theileria cervi (47.6%), Theileria sp. (14.3%), Babesia bovis (4.8%) and Babesia bigemina (4.8%). However, only one of the infected wild cervids exhibited accentuated anaemia (PCV=17%). This is first report concerning the occurrence of Theileria spp. in Brazilian cervids.

  20. Detection of rickettsiae in fleas and ticks from areas of Costa Rica with history of spotted fever group rickettsioses.

    PubMed

    Troyo, Adriana; Moreira-Soto, Rolando D; Calderon-Arguedas, Ólger; Mata-Somarribas, Carlos; Ortiz-Tello, Jusara; Barbieri, Amália R M; Avendaño, Adrián; Vargas-Castro, Luis E; Labruna, Marcelo B; Hun, Laya; Taylor, Lizeth

    2016-10-01

    Outbreaks of spotted fevers have been reported in Costa Rica since the 1950s, although vectors responsible for transmission to humans have not been directly identified. In this study, species of Rickettsia were detected in ectoparasites from Costa Rica, mostly from five study sites where cases of spotted fevers have been reported. Ticks and fleas were collected using drag cloths or directly from domestic and wild animals and pooled according to species, host, and location. Pools were analyzed initially by PCR to detect a fragment of Rickettsia spp. specific gltA gene, and those positive were confirmed by detection of htrA and/or ompA gene fragments. Partial sequences of the gltA gene were obtained, as well as at least one ompA and/or ompB partial sequence of each species. Rickettsia spp. were confirmed in 119 of 497 (23.9%) pools of ticks and fleas analyzed. Rickettsia rickettsii was identified in one nymph of Amblyomma mixtum and one nymph of Amblyomma varium. Other rickettsiae present were 'Candidatus Rickettsia amblyommii' in A. mixtum, Amblyomma ovale, Dermacentor nitens, and Rhipicephalus sanguineus s. l.; Rickettsia bellii in Amblyomma sabanerae; Rickettsia felis in Ctenocephalides felis; and Rickettsia sp. similar to 'Candidatus R. asemboensis' in C. felis, Pulex simulans, A. ovale, and Rhipicephalus microplus. Results show the presence of rickettsiae in vectors that may be responsible for transmission to humans in Costa Rica, and evidence suggests exposure to rickettsial organisms in the human environment may be common. This is the first study to report R. rickettsii in A. varium and in A. mixtum in Costa Rica.

  1. Ectoparasites of livestock, dogs, and wild rodents in the Chittagong Hill Tracts in southeastern Bangladesh.

    PubMed

    Fuehrer, Hans-Peter; Igel, Petra; Treiber, Moritz; Baumann, Timo A; Riedl, Julia; Swoboda, Paul; Joachim, Anja; Noedl, Harald

    2012-10-01

    A large variety of ectoparasites parasitizing on livestock, dogs, and rodents are documented throughout the world, of which several are proven vectors for major (including zoonotic) diseases affecting humans and/or livestock. However, there remains a significant lack of knowledge in regard to the ectoparasite fauna in remote regions of the developing world, such as southeastern Bangladesh, and an urgent need to investigate this fauna to improve diagnostic options. In the course of the present study, more than 5,300 ectoparasites were collected by flag dragging and handpicking of livestock, dogs, and rodents in the District of Bandarban (Chittagong Hill Tracts) in southeastern Bangladesh. Three tick species were identified: Haemaphysalis bispinosa (flagging, cattle, goats, and dogs), Rhipicephalus microplus (cattle, goats), and Rhipicephalus sanguineus (dogs, goats, and flagging). H. bispinosa was the dominant tick species on mammalian hosts as well as on vegetation. Furthermore, Ctenocephalides canis (dogs, goats) and Linognatus sp. (goat) were found. Overall, 73 rodents of eight different species (e.g., Mus musculus, Rattus sikkimensis, Bandicota bengalensis, and Niviventer sp.) hosted a variety of ectoparasites such as mites (Laelaps nuttali, Laelaps echidninus, Lyponissoides sp. and Ornithonyssus bacoti), fleas (Xenopsylla cheopis), and one myiasis-causing dipteran species. Monitoring the ectoparasite burden of livestock and other mammals is urgently needed in order to control ectoparasites associated with social and economic burden (e.g., reduced milk production, weight loss). Several zoonotic diseases can be transmitted by ectoparasites in this area, where the majority of the population live in basic housing conditions and in direct contact with livestock, dogs, and rodents.

  2. Modeling the Impact of Climate and Landscape on the Efficacy of White Tailed Deer Vaccination for Cattle Tick Control in Northeastern Mexico

    PubMed Central

    Estrada-Peña, Agustín; Carreón, Diana; Almazán, Consuelo; de la Fuente, José

    2014-01-01

    Cattle ticks are distributed worldwide and affect animal health and livestock production. White tailed deer (WTD) sustain and spread cattle tick populations. The aim of this study was to model the efficacy of anti-tick vaccination of WTD to control tick infestations in the absence of cattle vaccination in a territory where both host species coexist and sustain cattle tick populations. Agent-based models that included land cover/landscape properties (patch size, distances to patches) and climatic conditions were built in a GIS environment to simulate WTD vaccine effectiveness under conditions where unvaccinated cattle shared the landscape. Published and validated information on tick life cycle was used to build models describing tick mortality and developmental rates. Data from simulations were applied to a large territory in northeastern Mexico where cattle ticks are endemic and WTD and cattle share substantial portions of the habitat. WTD movements were simulated together with tick population dynamics considering the actual landscape and climatic features. The size of the vegetation patches and the distance between patches were critical for the successful control of tick infestations after WTD vaccination. The presence of well-connected, large vegetation patches proved essential for tick control, since the tick could persist in areas of highly fragmented habitat. The continued application of one yearly vaccination on days 1-70 for three years reduced tick abundance/animal/patch by a factor of 40 and 60 for R. annulatus and R. microplus, respectively when compared to non-vaccinated controls. The study showed that vaccination of WTD alone during three consecutive years could result in the reduction of cattle tick populations in northeastern Mexico. Furthermore, the results of the simulations suggested the possibility of using vaccines to prevent the spread and thus the re-introduction of cattle ticks into tick-free areas. PMID:25047078

  3. Non-Invasive Delivery of dsRNA into De-Waxed Tick Eggs by Electroporation.

    PubMed

    Ruiz, Newton; de Abreu, Leonardo Araujo; Parizi, Luís Fernando; Kim, Tae Kwon; Mulenga, Albert; Braz, Gloria Regina Cardoso; Vaz, Itabajara da Silva; Logullo, Carlos

    2015-01-01

    RNA interference-mediated gene silencing was shown to be an efficient tool for validation of targets that may become anti-tick vaccine components. Here, we demonstrate the application of this approach in the validation of components of molecular signaling cascades, such as the Protein Kinase B (AKT)/Glycogen Synthase Kinase (GSK) axis during tick embryogenesis. It was shown that heptane and hypochlorite treatment of tick eggs can remove wax, affecting corium integrity and but not embryo development. Evidence of AKT and GSK dsRNA delivery into de-waxed eggs of via electroporation is provided. Primers designed to amplify part of the dsRNA delivered into the electroporated eggs dsRNA confirmed its entry in eggs. In addition, it was shown that electroporation is able to deliver the fluorescent stain, 4',6-diamidino-2-phenylindole (DAPI). To confirm gene silencing, a second set of primers was designed outside the dsRNA sequence of target gene. In this assay, the suppression of AKT and GSK transcripts (approximately 50% reduction in both genes) was demonstrated in 7-day-old eggs. Interestingly, silencing of GSK in 7-day-old eggs caused 25% reduction in hatching. Additionally, the effect of silencing AKT and GSK on embryo energy metabolism was evaluated. As expected, knockdown of AKT, which down regulates GSK, the suppressor of glycogen synthesis, decreased glycogen content in electroporated eggs. These data demonstrate that electroporation of de-waxed R. microplus eggs could be used for gene silencing in tick embryos, and improve the knowledge about arthropod embryogenesis.

  4. Development of a multilocus sequence typing scheme for the study of Anaplasma marginale population structure over space and time.

    PubMed

    Guillemi, Eliana C; Ruybal, Paula; Lia, Verónica; Gonzalez, Sergio; Lew, Sergio; Zimmer, Patricia; Lopez Arias, Ludmila; Rodriguez, Jose L; Rodriguez, Sonia Y; Frutos, Roger; Wilkowsky, Silvina E; Farber, Marisa D

    2015-03-01

    Bovine Anaplasmosis caused by Anaplasma marginale is a worldwide disease prevalent in tropical and subtropical regions where Rhipicephalus microplus is considered the most significant biological vector. Molecular markers previously applied for A. marginale typing are efficient for isolate discrimination but they are not a suitable tool for studying population structure and dynamics. Here we report the development of an MLST scheme based on the study of seven genes: dnaA, ftsZ, groEl, lipA, recA, secY and sucB. Five annotated genomes (Saint Maries, Florida, Mississippi, Puerto Rico and Virginia) and 53 bovine blood samples from different world regions were analyzed. High nucleotide diversity and a large proportion of synonymous substitutions, indicative of negative selection resulted from DnaSP 5.00.02 package application. Recombination events were detected in almost all genes, this evidence together with the coexistence of more than one A. marginale strain in the same sample might suggest the superinfection phenomena as a potential source of variation. The allelic profile analysis performed through GoeBURST shown two main CC that did not support geography. In addition, the AMOVA test confirmed the occurrence of at least two main genetically divergent groups. The composition of the emergent groups reflected the impact of both historical and environmental traits on A. marginale population structure. Finally, a web-based platform "Galaxy MLST-Pipeline" was developed to automate DNA sequence editing and data analysis that together with the Data Base are freely available to users. The A. marginale MLST scheme developed here is a valuable tool with a high discrimination power, besides PCR based strategies are still the better choice for epidemiological intracellular pathogens studies. Finally, the allelic profile describe herein would contribute to uncover the mechanisms in how intracellular pathogens challenge virulence paradigm.

  5. Morphological and genetic diversity of Rhipicephalus sanguineus sensu lato from the New and Old Worlds

    PubMed Central

    2013-01-01

    Background The taxonomic status of the brown dog tick (Rhipicephalus sanguineus sensu stricto), which has long been regarded as the most widespread tick worldwide and a vector of many pathogens to dogs and humans, is currently under dispute. Methods We conducted a comprehensive morphological and genetic study of 278 representative specimens, which belonged to different species (i.e., Rhipicephalus bursa, R. guilhoni, R. microplus, R. muhsamae, R. pusillus, R. sanguineus sensu lato, and R. turanicus) collected from Europe, Asia, Americas, and Oceania. After detailed morphological examination, ticks were molecularly processed for the analysis of partial mitochondrial (16S rDNA, 12S rDNA, and cox1) gene sequences. Results In addition to R. sanguineus s.l. and R. turanicus, three different operational taxonomic units (namely, R. sp. I, R. sp. II, and R. sp. III) were found on dogs. These operational taxonomical units were morphologically and genetically different from R. sanguineus s.l. and R. turanicus. Ticks identified as R. sanguineus s.l., which corresponds to the so-called “tropical species” (=northern lineage), were found in all continents and genetically it represents a sister group of R. guilhoni. R. turanicus was found on a wide range of hosts in Italy and also on dogs in Greece. Conclusions The tropical species and the temperate species (=southern lineage) are paraphyletic groups. The occurrence of R. turanicus in the Mediterranean region is confirmed. A consensual re-description of R. sanguineus s.s. and R. turanicus will be necessary to solve the taxonomic problems within the so-called R. sanguineus group. PMID:23880226

  6. Non-Invasive Delivery of dsRNA into De-Waxed Tick Eggs by Electroporation

    PubMed Central

    Ruiz, Newton; de Abreu, Leonardo Araujo; Parizi, Luís Fernando; Kim, Tae Kwon; Mulenga, Albert; Braz, Gloria Regina Cardoso; Vaz, Itabajara da Silva; Logullo, Carlos

    2015-01-01

    RNA interference-mediated gene silencing was shown to be an efficient tool for validation of targets that may become anti-tick vaccine components. Here, we demonstrate the application of this approach in the validation of components of molecular signaling cascades, such as the Protein Kinase B (AKT) / Glycogen Synthase Kinase (GSK) axis during tick embryogenesis. It was shown that heptane and hypochlorite treatment of tick eggs can remove wax, affecting corium integrity and but not embryo development. Evidence of AKT and GSK dsRNA delivery into de-waxed eggs of via electroporation is provided. Primers designed to amplify part of the dsRNA delivered into the electroporated eggs dsRNA confirmed its entry in eggs. In addition, it was shown that electroporation is able to deliver the fluorescent stain, 4',6-diamidino-2-phenylindole (DAPI). To confirm gene silencing, a second set of primers was designed outside the dsRNA sequence of target gene. In this assay, the suppression of AKT and GSK transcripts (approximately 50% reduction in both genes) was demonstrated in 7-day-old eggs. Interestingly, silencing of GSK in 7-day-old eggs caused 25% reduction in hatching. Additionally, the effect of silencing AKT and GSK on embryo energy metabolism was evaluated. As expected, knockdown of AKT, which down regulates GSK, the suppressor of glycogen synthesis, decreased glycogen content in electroporated eggs. These data demonstrate that electroporation of de-waxed R. microplus eggs could be used for gene silencing in tick embryos, and improve the knowledge about arthropod embryogenesis. PMID:26091260

  7. Rickettsial Infection in Animals, Humans and Ticks in Paulicéia, Brazil.

    PubMed

    Silveira, I; Martins, T F; Olegário, M M; Peterka, C; Guedes, E; Ferreira, F; Labruna, M B

    2015-11-01

    A previous study in Paulicéia Municipality, south-eastern Brazil, reported 9.7% of the Amblyomma triste ticks to be infected by Rickettsia parkeri, a bacterial pathogen that causes spotted fever in humans. These A. triste ticks were shown to be associated with marsh areas, where the marsh deer Blastocerus dichotomus is a primary host for this tick species. During 2008-2009, blood serum samples were collected from 140 horses, 41 dogs, 5 opossums (Didelphis albiventris) and 26 humans in farms from Pauliceia Municipality. Ticks were collected from these animals, from vegetation and from additional wildlife in these farms. Overall, 25% (35/140) of the horses, 7.3% (3/41) of the dogs, 3.8% (1/26) of the humans and 100% (5/5) of the opossums were seroreactive (titre ≥64) to spotted fever group (SFG) Rickettsia spp. Multivariate statistical analysis indicated that horses that were allowed to forage in the marsh were 4.8 times more likely to be seroreactive to spotted fever group (SFG) Rickettsia spp than horses that did not forage in the marsh. In addition, horses that had been living in the farm for more than 8.5 years were 2.8 times more likely to be seroreactive to SFG Rickettsia spp than horses that were living for ≤8.5 years. Ticks collected from domestic animals or from vegetation included Amblyomma cajennense, Amblyomma coelebs, Amblyomma dubitatum, Dermacentor nitens and Rhipicephalus microplus. By PCR analyses, only one pool of A. coelebs ticks from the vegetation was shown to be infected by rickettsiae, for which DNA sequencing revealed to be Rickettsia amblyommii. Ticks (not tested by PCR) collected from wildlife encompassed A. cajennense and Amblyomma rotundatum on lizards (Tupinambis sp), and A. cajennense and A. triste on the bird Laterallus viridis. Our results indicate that the marsh area of Paulicéia offers risks of infection by SFG rickettsiae.

  8. Molecular and structural characterization of novel cystatins from the taiga tick Ixodes persulcatus.

    PubMed

    Rangel, Carolina K; Parizi, Luís F; Sabadin, Gabriela A; Costa, Evenilton P; Romeiro, Nelilma C; Isezaki, Masayoshi; Githaka, Naftaly W; Seixas, Adriana; Logullo, Carlos; Konnai, Satoru; Ohashi, Kazuhiko; da Silva Vaz, Itabajara

    2017-03-01

    Cystatins are cysteine peptidase inhibitors that in ticks mediate processes such as blood feeding and digestion. The ixodid tick Ixodes persulcatus is endemic to the Eurasia, where it is the principal vector of Lyme borreliosis. To date, no I. persulcatus cystatin has been characterized. In the present work, we describe three novel cystatins from I. persulcatus, named JpIpcys2a, JpIpcys2b and JpIpcys2c. In addition, the potential of tick cystatins as cross-protective antigens was evaluated by vaccination of hamsters using BrBmcys2c, a cystatin from Rhipicephalus microplus, against I. persulcatus infestation. Sequence analysis showed that motifs that are characteristic of cystatins type 2 are fully conserved in JpIpcys2b, while mutations are present in both JpIpcys2a and JpIpcys2c. Protein-protein docking simulations further revealed that JpIpcys2a, JpIpcys2b and JpIpcys2c showed conserved binding sites to human cathepsins L, all of them covering the active site cleft. Cystatin transcripts were detected in different I. persulcatus tissues and instars, showing their ubiquitous expression during I. persulcatus development. Serological analysis showed that although hamsters immunized with BrBmcys2c developed a humoral immune response, this response was not adequate to protect against a heterologous challenge with I. persulcatus adult ticks. The lack of cross-protection provided by BrBmcys2c immunization is perhaps linked to the fact that cystatins cluster into multigene protein families that are expressed differentially and exhibit functional redundancy. How to target such small proteins that are secreted in low quantities remains a challenge in the development of suitable anti-tick vaccine antigens.

  9. Epidemiological perspectives of ticks and tick-borne diseases in South Sudan: cross-sectional survey results.

    PubMed

    Kivaria, Fredrick M; Kapaga, Angolwisye M; Mbassa, Gabriel K; Mtui, Paul F; Wani, Rhombe J

    2012-09-03

    A cross-sectional study was conducted between September and October 2010 in five states of South Sudan that were selected on the basis of the perceived risk of tick-borne diseases. The purpose was to investigate epidemiological parameters of tick-borne diseases in South Sudan and their uses in future control strategies. A total of 805 calves were assessed by clinical, microscopic and serological examination and tick counts. The indirect Enzyme-Linked Immuno-Sorbent Assay (ELISA) was used to detect antibodies to Theileria parva, Theileria mutans, Anaplasma marginale and Babesian bigemina. Sero-conversion risks for T. parva and T. mutans were 27.3% and 31.3% respectively, whilst the risk was 57.6% and 52.8% for A. marginale and B. bigemina, respectively. Major tick species identified include Rhipicephalus appendiculatus, Rhipicephalus decoloratus, Rhipicephalus microplus, Amblyomma variegatum, and Rhipicephalus evertsi. There was great variation (P ≤ 0.001) in the number of all these ticks, both between herds in a state and between calves in an individual herd. The low and intermediate sero-conversion risks observed in the study states suggest that immunisation against East Coast fever (ECF) is justified. Fortunately, three major genotypes that were identified by applying Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCRRFLP) analysis on the p104 to the blood samples and T. parva Muguga, matched very well with T. parva Kiambu 5 and T. parva Muguga; therefore the Muguga cocktail can be used for the immunisation of cattle in South Sudan. However, prospective studies are required to develop optimal control measures for tick-borne diseases under different ecological and husbandry practices in South Sudan.

  10. Developing Anti-tick Vaccines.

    PubMed

    Rodríguez-Mallon, Alina

    2016-01-01

    Ticks are responsible for the transmission of viral, bacterial, and protozoal diseases of man and animals and also produce significant economic losses to cattle industry. The use of acaricides constitutes a major component of integrated tick control strategies. However, this is accompanied by the selection of acaricide-resistant ticks and contamination of environment and milk and meat products with drug residues. These issues highlight the need for alternative approaches to control tick infestations and have triggered the search for tick protective antigens for vaccine development. Vaccination as a tick control method has been practiced since the introduction of TickGARD and Gavac that were developed using the midgut glycoprotein Bm86 as antigen. Gavac within integrated tick management systems has proven to reduce the number of acaricidal applications per year that are required to control some strains of R. microplus ticks in different geographical regions. Nevertheless, it has limited or no efficacy against other tick species. These issues have stimulated research for additional tick protective antigens with critical functions in the tick. This chapter presents methodologies for the design and test of molecules as antigens against ticks. Considerations about different methods for the tick control compared to the immunological methods, the desirable characteristics for an anti-tick vaccine and the obstacles encountered for developing this kind of vaccines are discussed. Detailed methodologies for the establishment of a biological model to test new molecules as immunogens against ticks and to perform challenge trials with this model are presented. General considerations in the efficacy calculation for any anti-tick vaccine are also discussed.

  11. Structural imprints at the front of the Chocó-Panamá indenter: Field data from the North Cauca Valley Basin, Central Colombia

    NASA Astrophysics Data System (ADS)

    Suter, F.; Sartori, M.; Neuwerth, R.; Gorin, G.

    2008-11-01

    The northern Andes are a complex area where tectonics is dominated by the interaction between three major plates and accessory blocks, in particular, the Chocó-Panamá and Northern Andes Blocks. The studied Cauca Valley Basin is located at the front of the Chocó-Panamá Indenter, where the major Romeral Fault System, active since the Cretaceous, changes its kinematics from right-lateral in the south to left-lateral in the north. Structural studies were performed at various scales: DEM observations in the Central Cordillera between 4 and 5.7°N, aerial photograph analyses, and field work in the folded Oligo-Miocene rocks of the Serranía de Santa Barbara and in the flat-lying, Pleistocene Quindío-Risaralda volcaniclastic sediments interfingering with the lacustrine to fluviatile sediments of the Zarzal Formation. The data acquired allowed the detection of structures with a similar orientation at every scale and in all lithologies. These families of structures are arranged similarly to Riedel shears in a right-lateral shear zone and are superimposed on the Cretaceous Romeral suture. They appear in the Central Cordillera north of 4.5°N, and define a broad zone where 060-oriented right-lateral distributed shear strain affects the continental crust. The Romeral Fault System stays active and strain partitioning occurs among both systems. The southern limit of the distributed shear strain affecting the Central Cordillera corresponds to the E-W trending Garrapatas-Ibagué shear zone, constituted by several right-stepping, en-échelon, right-lateral, active faults and some lineaments. North of this shear zone, the Romeral Fault System strike changes from NNE to N. Paleostress calculations gave a WNW-ESE trending, maximum horizontal stress, and 69% of compressive tensors. The orientation of σ1 is consistent with the orientation of the right-lateral distributed shear strain and the compressive state characterizing the Romeral Fault System in the area: it bisects the

  12. Ectoparasites of sheep in three agro-ecological zones in central Oromia, Ethiopia.

    PubMed

    Kumsa, Bersissa; Beyecha, Kebede; Geloye, Mesula

    2012-10-23

    A cross-sectional study was conducted to determine the prevalence and risk factors for ectoparasites infestation in sheep in three agro-ecological zones in central Oromia, Ethiopia, from October 2009 to April 2010. The study revealed that 637 (48.1%) of the 1325 sheep examined were infested with one or more ectoparasites. The ectoparasites identified were Bovicola ovis (27.2%), Melophagus ovinus (16.4%), Ctenocephalides sp. (2.3%), Linognathus africanus (1.2%), Linognathus ovillus (0.3%), Sarcoptes sp. (1.2%), Amblyomma variegatum (4.4%), Rhipicephalus evertsi evertsi (1.9%), Rhipicephalus pravus (1.9%), Rhipicephalus (Boophilus) decoloratus (1.1%), Rhipicephalus sanguineus (0.9%), Rhipicephalus praetextatus (1.1%) and Hyalomma truncatum (1.6%). Statistically significant difference was observed in prevalence of B. ovis amongst study agroecological zones: highland 36.6%, midland 20.9% and lowland 14.0%. Significantly higher prevalence was recorded in highland agroecological zone. A significantly (OR = 0.041, p < 0.001) higher prevalence of M. ovinus in the highland (31.7%) than in both the lowland (0%) and midland (1.9%) was observed. The risk of tick infestation in the lowland and midland was 9.883 times and 13.988 times higher than the risk in the highland, respectively. A significantly higher prevalence of Ctenocephalides species was encountered in both the lowland (OR = 4.738, p = 0.011) and midland (OR = 8.078, p = 0.000) than in the highland agro-ecological zone. However, a significant difference (p = 0.191) amongst agro-ecological zones was not found for the prevalence of Linognathus and Sarcoptes species. Statistically significant variation (p > 0.05) was never recorded in the prevalence of all the identified species of ectoparasites between male and female sheep hosts. However, a significantly (p = 0.006) higher prevalence of B. ovis was recorded between young and adult sheep. The risk of B. ovis infestation was 1.45 times higher in young than the adult

  13. Zoonotic surveillance for rickettsiae in domestic animals in Kenya.

    PubMed

    Mutai, Beth K; Wainaina, James M; Magiri, Charles G; Nganga, Joseph K; Ithondeka, Peter M; Njagi, Obadiah N; Jiang, Ju; Richards, Allen L; Waitumbi, John N

    2013-06-01

    Abstract Rickettsiae are obligate intracellular bacteria that cause zoonotic and human diseases. Arthropod vectors, such as fleas, mites, ticks, and lice, transmit rickettsiae to vertebrates during blood meals. In humans, the disease can be life threatening. This study was conducted amidst rising reports of rickettsioses among travelers to Kenya. Ticks and whole blood were collected from domestic animals presented for slaughter at major slaughterhouses in Nairobi and Mombasa that receive animals from nearly all counties in the country. Blood samples and ticks were collected from 1019 cattle, 379 goats, and 299 sheep and were screened for rickettsiae by a quantitative PCR (qPCR) assay (Rick17b) using primers and probe that target the genus-specific 17-kD gene (htrA). The ticks were identified using standard taxonomic keys. All Rick17b-positive tick DNA samples were amplified and sequenced with primers sets that target rickettsial outer membrane protein genes (ompA and ompB) and the citrate-synthase encoding gene (gltA). Using the Rick17b qPCR, rickettsial infections in domestic animals were found in 25/32 counties sampled (78.1% prevalence). Infection rates were comparable in cattle (16.3%) and sheep (15.1%) but were lower in goats (7.1%). Of the 596 ticks collected, 139 had rickettsiae (23.3%), and the detection rates were highest in Amblyomma (62.3%; n=104), then Rhipicephalus (45.5%; n=120), Hyalomma (35.9%; n=28), and Boophilus (34.9%; n=30). Following sequencing, 104 out of the 139 Rick17b-positive tick DNA had good reverse and forward sequences for the 3 target genes. On querying GenBank with the generated consensus sequences, homologies of 92-100% for the following spotted fever group (SFG) rickettsiae were identified: Rickettsia africae (93.%, n=97), Rickettsia aeschlimannii (1.9%, n=2), Rickettsia mongolotimonae (0.96%, n=1), Rickettsia conorii subsp. israelensis (0.96%, n=1), Candidatus Rickettsia kulagini (0.96% n=1), and Rickettsia spp. (1.9% n=2). In

  14. New Borrelia species detected in ixodid ticks in Oromia, Ethiopia.

    PubMed

    Kumsa, Bersissa; Socolovschi, Cristina; Raoult, Didier; Parola, Philippe

    2015-04-01

    Little is known about Borrelia species transmitted by hard ticks in Ethiopia. The present study was conducted from November 2011 through March 2014 to address the occurrence and molecular identity of these bacteria in ixodid ticks infesting domestic animals in Oromia, Ethiopia. A total of 767 ixodid ticks collected from domestic animals were screened for Borrelia DNA by quantitative (q) real-time PCR followed by standard PCR and sequencing to identify the species. Overall, 3.8% (29/767) of the tested ticks were positive for Borrelia DNA, including 8/119 (6.7%) Amblyomma cohaerens, 1/42 (2.4%) Am. gemma, 3/53 (5.7%) Am. variegatum, 5/22 (22.7%) Amblyomma larvae, 3/60 (5%) Amblyomma nymphs, 2/139 (1.4%) Rhipicephalus (Boophilus) decoloratus, 2/31 (6.4%) Rh. decoloratus nymphs, and 5/118 (4.2%) Rh. pulchellus using 16S genus-specific qPCR. The prevalence of Borrelia DNA was significantly higher in genus Amblyomma (20/298, 6.7%) than in the genus Rhipicephalus (9/417, 2.1%) ticks (P=0.001). Sequencing of PCR products from the flaB and 16S rRNA genes of Borrelia spp. from Amblyomma ticks showed the presence of a new species between the relapsing fever and Lyme disease groups. However, Borrelia sp. detected in Rhipicephalus ticks clustered with B. theileri/B. lonestari. The human pathogenicity of the Borrelia sp. detected in Amblyomma ticks from Ethiopia has not yet been investigated, whereas the Borrelia sp. detected in Rhipicephalus ticks in our study is the causative agent of bovine borreliosis in cattle and may have veterinary importance in different parts of Ethiopia. Furthermore, the detection of previously unrecognized Borrelia species in Amblyomma and Rhipicephalus ticks in Ethiopia generates additional questions concerning the bacterial fauna in hard ticks and will prompt researchers to perform detailed studies for better understanding of ixodid ticks associated bacteria.

  15. Acetylcholinesterases of blood-feeding flies and ticks.

    PubMed

    Temeyer, Kevin B; Tuckow, Alexander P; Brake, Danett K; Li, Andrew Y; Pérez de León, Adalberto A

    2013-03-25

    Acetylcholinesterase (AChE) is the biochemical target of organophosphate (OP) and carbamate pesticides for invertebrates, vertebrate nerve agents, and AChE inhibitors used to reduce effects of Alzheimer's disease. Organophosphate pesticides (OPs) are widely used to control blood-feeding arthropods, including biting flies and ticks. However, resistance to OPs in pests affecting animal and human health has compromised control efficacy. OP resistance often results from mutations producing an OP-insensitive AChE. Our studies have demonstrated production of OP-insensitive AChEs in biting flies and ticks. Complementary DNA (cDNA) sequences encoding AChEs were obtained for the horn fly, stable fly, sand fly, and the southern cattle tick. The availability of cDNA sequences enables the identification of mutations, expression and characterization of recombinant proteins, gene silencing for functional studies, as well as in vitro screening of novel inhibitors. The southern cattle tick expresses at least three different genes encoding AChE in their synganglion, i.e. brain. Gene amplification for each of the three known cattle tick AChE genes and expression of multiple alleles for each gene may reduce fitness cost associated with OP-resistance. AChE hydrolyzes the neurotransmitter, acetylcholine, but may have additional roles in physiology and development. The three cattle tick AChEs possess significantly different biochemical properties, and are expressed in neural and non-neural tissues, which suggest separation of structure and function. The remarkable complexity of AChEs in ticks suggested by combining genomic data from Ixodes scapularis with our genetic and biochemical data from Rhipicephalus microplus is suggestive of previously unknown gene duplication and diversification. Comparative studies between invertebrate and vertebrate AChEs could enhance our understanding of structure-activity relationships. Research with ticks as a model system offers the opportunity to

  16. Resistance of beef cattle of two genetic groups to ectoparasites and gastrointestinal nematodes in the state of São Paulo, Brazil.

    PubMed

    Oliveira, M C S; Alencar, M M; Giglioti, R; Beraldo, M C D; Aníbal, F F; Correia, R O; Boschini, L; Chagas, A C S; Bilhassi, T B; Oliveira, H N

    2013-10-18

    The resistance to infestations by ectoparasites and infections by gastrointestinal nematodes was studied in 45 animals (males and females) of two genetic groups: purebred Nelore (NI, n=28) and Three-Cross (1/2 Angus+1/4 Canchim+1/4 Nelore - TC, n=17). The animals were monitored for 24 months, during which they were left to graze in tropical pastures without receiving treatment for parasites. Each month the animals were examined for infestations by external parasites, to count the numbers of cattle ticks Rhipicephalus microplus with diameter greater than 4.5mm present on the left side, horn flies (Haematobia irritans) present in the lumbar region and botfly larvae (Dermatobia hominis) present on the entire body. The H. irritans counts were performed with the aid of digital photographs. At the time of examination, fecal samples were collected to count the eggs per gram (EPG) and to perform coprocultures, and peripheral blood samples were drawn to determine the packed cell volume (PCV) and to count the eosinophils. For statistical analysis, the count data were transformed into log₁₀ (n+1), where n is the number of parasites. For PCV, significant effects (P<0.05) were found for collection month (CO), genetic group (GG) and gender (SX), with means and respective standard errors of 41.5 ± 0.65% for the NI animals, 39.3 ± 0.83% for the TC, 41.5 ± 0.72% for the females and 39.3 ± 0.77% for the males. Regarding the eosinophil counts, only the effect of sex was significant (P<0.01), with means and respective standard errors of 926.0 ± 46.2/μL, for males and 1088.0 ± 43.8/μL of blood, for females. The NI animals presented lower mean counts for all the external parasites compared to the TC animals (P<0.01). For ticks, the transformed means followed by standard errors for the NI and TC animals were 0.06 ± 0.01 and 0.34 ± 0.02, while for horn flies these were 0.92 ± 0.05 and 1.36 ± 0.06 and for botfly larvae they were 0.05 ± 0.03 and 0.45 ± 0.05, respectively