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Sample records for garrapata boophilus microplus

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

    USDA-ARS?s Scientific Manuscript database

    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. Eprinomectin accumulation in Rhipicephalus (Boophilus) microplus: Pharmacokinetic and efficacy assessment.

    PubMed

    Lifschitz, A; Nava, S; Mangold, A; Imperiale, F; Ballent, M; Canevari, J; Lanusse, C

    2016-01-15

    Eprinomectin (EPM) is a macrocyclic lactone used against endo-ectoparasites without withdrawal time in milk and meat after its pour-on administration at 0.5mg/kg. Previous experiments evaluated the efficacy of EPM against Rhipicephalus (Boophilus) microplus in cattle. This study assessed EPM efficacy against R. (B.) microplus after topical administration at two dose rates and investigated the relationship between EPM systemic exposure in the host and drug concentrations accumulated in ticks recovered from treated animals. A standardized pharmaco-parasitological study was performed in two phases. In phase 1 eighteen Braford cattle naturally infected with R. (B.) microplus were divided into three experimental groups with a similar level of infestation (Kruskal-Wallis test, P>0.05): control group and treated groups with EPM pour-on (1 and 1.5mg/kg). Samples of heparinized blood and ticks at different life stages were taken between 0 and 21 days (d) post-administration to measure EPM concentrations by HPLC. The efficacy trial (phase 2) included eighteen Braford calves naturally infected with R. (B.) microplus divided into control group and 1mg/kg and 1.5mg/kg EPM treated groups. Female ticks (4.5-8mm) on cattle were counted between 1 and 23 days post-treatment to evaluate the efficacy of EPM. The reproductive efficiency index (REI) and the fertility efficiency index (FEI) were evaluated. Plasma concentrations of EPM showed a linear relationship with the level of dose rate administered. Peak plasma concentrations were within a range between 13.8 and 90ng/ml, which guarantee milk drug concentrations below the maximum residues level. High EPM concentrations were detected in ticks. EPM concentrations in R. (B.) microplus were correlated to plasma concentrations between 1.25 days and 21 days post-administration (r 0.84; P<0.05). EPM efficacy calculated using the Henderson-Tilton formula was 98.9% and 99.1% (7 days post-administration) and 100% (23 days post

  5. 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.

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

    USDA-ARS?s Scientific Manuscript database

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

    USDA-ARS?s Scientific Manuscript database

    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 da...

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

    USDA-ARS?s Scientific Manuscript database

    Six strains of Rhipicephalus (Boophilus) microplus collected from northern Mexico were surveyed for resistance to 5 classes of acaricide. All were resistant to permethrin. Two strains were resistant to amitraz and 3 were found to be resistant to coumaphos. Two strains were resistant to fipronil incl...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  11. Microarray analysis of acaricide inducible gene expression in the southern cattle tick, Rhipicephalus (Boophilus) microplus

    USDA-ARS?s Scientific Manuscript database

    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...

  12. Resistance of the tick Rhipicephalus (Boophilus) microplus to ivermectin in Mexico

    USDA-ARS?s Scientific Manuscript database

    Ticks and the diseases they transmit cause great economic losses to livestock in many tropical and sub-tropical countries, including Mexico. Chemical acaricides have been widely used to control the cattle tick, Rhipicephalus (Boophilus) microplus, in Mexico. Resistance to organophosphate (OP), synth...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

  15. Multiple acetylcholinesterase genes expressed in the southern cattle tick, Rhipicephalus (Boophilus) microplus, a vector of bovine anaplasmosis and babesiosis

    USDA-ARS?s Scientific Manuscript database

    Cattle ticks of the genus Boophilus (now Rhipicephalus) were eradicated from the United States, but remain endemic in Central and South America, as well as portions of Africa, Asia, and Australia. Rhipicephalus (Boophilus) microplus and annulatus ticks present a continuing threat to the U.S. cattle ...

  16. Multiple acetylcholinesterase genes expressed in the southern cattle tick, Rhipicephalus (Boophilus) microplus, a vector of bovine babesiosis and anaplasmosis

    USDA-ARS?s Scientific Manuscript database

    Cattle ticks of the genus Boophilus (now Rhipicephalus) were eradicated from the United States, but remain endemic in Central and South America, as well as portions of Africa, Asia, and Australia. Rhipicephalus (Boophilus) microplus and annulatus ticks present a continuing threat to the U.S. cattle ...

  17. A Novel Carbamate Insecticide with Superior Selectivity for Cattle Tick (Boophilus microplus) and Sand Fly (Phlebotomus papatasi) Control

    USDA-ARS?s Scientific Manuscript database

    The cattle tick, Boophilus microplus, and the sand fly, Phlebotomus papatasi (Pp), are vectors of infectious agents affecting cattle and humans, respectively. The purpose of this study was to characterize the inhibitor profile of acetylcholinesterases from R. microplus(BmAChE1) and Pp (PpAchE) for c...

  18. 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.

  19. Phylogenetic analysis of mitochondrial genome sequences indicates that the cattle tick, Rhipicephalus (Boophilus) microplus, contains a cryptic species.

    PubMed

    Burger, Thomas D; Shao, Renfu; Barker, Stephen C

    2014-07-01

    Cattle ticks of the subgenus Rhipicephalus (Boophilus) are major agricultural pests worldwide, causing billions of dollars in losses annually. Rhipicephalus (Boophilus) annulatus and R. microplus are the most well-known and widespread species, and a third species, R. australis, was recently reinstated for 'R. microplus' from Australia and parts of Southeast Asia. We use mitochondrial genome sequences to address the phylogenetic relationships among the species of the subgenus Boophilus. We sequenced the complete or partial mitochondrial genomes of R. annulatus, R. australis, R. kohlsi, R. geigyi, and of three geographically disparate specimens of R. microplus from Brazil, Cambodia and China. Phylogenetic analyses of mitochondrial genomes, as well as cox1 and 16S rRNA sequences, reveals a species complex of R. annulatus, R. australis, and two clades of R. microplus, which we call the R. microplus complex. We show that cattle ticks morphologically identified as R. microplus from Southern China and Northern India (R. microplus clade B) are more closely related to R. annulatus than other specimens of R. microplus s.s. from Asia, South America and Africa (R. microplus clade A). Our analysis suggests that ticks reported as R. microplus from Southern China and Northern India are a cryptic species. This highlights the need for further molecular, morphological and crossbreeding studies of the R. microplus complex, with emphasis on specimens from China and India. We found that cox1 and, to a lesser extent, 16S rRNA were far more successful in resolving the phylogenetic relationships within the R. microplus complex than 12S rRNA or the nuclear marker ITS2. We suggest that future molecular studies of the R. microplus complex should focus on cox1, supplemented by 16S rRNA, and develop nuclear markers alternative to ITS2 to complement the mitochondrial data. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. Efficacy of eprinomectin pour-on against Rhipicephalus (Boophilus) microplus on buffaloes.

    PubMed

    Nazir, T; Katoch, R; Godara, R; Yadav, Anish

    2013-10-01

    A trial was conducted on 12 buffaloes naturally infested with Rhipicephalus (Boophilus) microplus to evaluate the efficacy of eprinomectin pour-on at a dose of 0.5 mg/kg body weight. A reduction in live tick count by 45.94, 63.96, 81.53, 90.54, 98.19 and 100 % was observed on days 3, 5, 7, 10, 14 and 21 post-treatment, respectively. The reinfestation of ticks was not observed up to 42 days of trial period. On the basis of the present trial of eprinomectin pour-on, it can be recommended for use in dairy buffaloes against Rhipicephalus (Boophilus) microplus infestation.

  1. Bovine immunoprotection against Rhipicephalus (Boophilus) microplus with recombinant Bm86-Campo Grande antigen.

    PubMed

    Cunha, Rodrigo Casquero; Pérez de León, Adalberto Angel; Leite, Fábio Pereira Leivas; Pinto, Luciano da Silva; Dos Santos Júnior, Alceu Gonçalves; Andreotti, Renato

    2012-01-01

    The southern cattle fever tick, Rhipicephalus (Boophilus) microplus, is no doubt the most economically important ectoparasite of cattle globally. The inappropriate use of chemical acaricides has driven the evolution of resistance in populations of R. (B.) microplus. Anti-tick vaccines represent a technology that can be combined with acaricides in integrated control programs to mitigate the impact of R. (B.) microplus. The recombinant form of Bm86 antigen from the Campo Grande (rBm86-CG) strain of R. (B.) microplus was produced using the Pichiapastoris expression system to test its ability to immunoprotect cattle against tick infestation. Secretion of rBm86-CG by P. pastoris through the bioprocess reported here simplified purification of the antigen. A specific humoral immune response was detected by ELISA in vaccinated cattle. Immunoblot results revealed that polyclonal antibodies from vaccinated cattle recognized a protein in larval extracts with a molecular weight corresponding to Bm86. The rBm86-CG antigen showed 31% efficacy against the Campo Grande strain of R. (B.) microplus infesting vaccinated cattle. The rBm86-CG is an antigen that could be used in a polyvalent vaccine as part of an integrated program for the control of R. (B.) microplus in the region that includes Mato Grosso do Sul.

  2. 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.

  3. 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.

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

    USDA-ARS?s Scientific Manuscript database

    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. ...

  5. 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.

  6. 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.

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

    PubMed

    Guerrero, Felix D; Andreotti, Renato; Bendele, Kylie G; Cunha, Rodrigo C; Miller, Robert J; Yeater, Kathleen; Pérez de León, Adalberto A

    2014-10-12

    Vaccination as a control method against the cattle tick, Rhipicephalus (Boophilus) microplus has been practiced since the introduction of two products in the mid-1990s. There is a need for a vaccine that could provide effective control of R. microplus in a more consistent fashion than existing products. During our transcriptome studies of R. microplus, several gene coding regions were discovered to encode proteins with significant amino acid similarity to aquaporins. A cDNA encoding an aquaporin from the cattle tick, Rhipicephalus microplus, was isolated from transcriptomic studies conducted on gut tissues dissected from fully engorged adult female R. microplus. 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 channels can also facilitate the transport of glycerol in addition to water. The efficacy of this aquaporin as an antigen against the cattle tick was explored in cattle vaccine trials conducted in Brazil. A cDNA encoding a significant portion of RmAQP1 was expressed as a recombinant protein in Pichia pastoris, purified under native conditions using a polyhistidine C-terminus tag and nickel affinity chromatography, emulsified with Montanide adjuvant, and cattle vaccinated intramuscularly. The recombinant protein provided 75% and 68% efficacy in two cattle pen trials conducted in Campo Grande, Brazil on groups of 6 one year old Holstein calves. The effectiveness of this vaccine in reducing the numbers of adult female ticks shows this aquaporin antigen holds promise as an active ingredient in cattle vaccines targeted against infestations of R. microplus.

  8. Global comparative analysis of ESTs from the southern cattle tick, Rhipicephalus (Boophilus) microplus

    PubMed Central

    Wang, Minghua; Guerrero, Felix D; Pertea, Geo; Nene, Vishvanath M

    2007-01-01

    Background The southern cattle tick, Rhipicephalus (Boophilus) microplus, is an economically important parasite of cattle and can transmit several pathogenic microorganisms to its cattle host during the feeding process. Understanding the biology and genomics of R. microplus is critical to developing novel methods for controlling these ticks. Results We present a global comparative genomic analysis of a gene index of R. microplus comprised of 13,643 unique transcripts assembled from 42,512 expressed sequence tags (ESTs), a significant fraction of the complement of R. microplus genes. The source material for these ESTs consisted of polyA RNA from various tissues, lifestages, and strains of R. microplus, including larvae exposed to heat, cold, host odor, and acaricide. Functional annotation using RPS-Blast analysis identified conserved protein domains in the conceptually translated gene index and assigned GO terms to those database transcripts which had informative BlastX hits. Blast Score Ratio and SimiTri analysis compared the conceptual transcriptome of the R. microplus database to other eukaryotic proteomes and EST databases, including those from 3 ticks. The most abundant protein domains in BmiGI were also analyzed by SimiTri methodology. Conclusion These results indicate that a large fraction of BmiGI entries have no homologs in other sequenced genomes. Analysis with the PartiGene annotation pipeline showed 64% of the members of BmiGI could not be assigned GO annotation, thus minimal information is available about a significant fraction of the tick genome. This highlights the important insights in tick biology which are likely to result from a tick genome sequencing project. Global comparative analysis identified some tick genes with unexpected phylogenetic relationships which detailed analysis attributed to gene losses in some members of the animal kingdom. Some tick genes were identified which had close orthologues to mammalian genes. Members of this group

  9. 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

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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 resistanc...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  19. Cattle Fever Tick, Rhipicephalus (Boophilus) microplus, (Acari: Ixodidae): potential control on pastures by the application of urea fertilizer

    USDA-ARS?s Scientific Manuscript database

    The southern cattle fever tick, Rhipicephalus (Boophilus) microplus, spends as much as 80–90% of its life cycle as a larva questing for a host. Standard control methods are limited to on-host applications, leaving a need for methods directed at the pasture infesting stages. Reports from Brazil indic...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  1. 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

  2. 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

    USDA-ARS?s Scientific Manuscript database

    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 ...

  3. 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

    USDA-ARS?s Scientific Manuscript database

    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...

  4. 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

    USDA-ARS?s Scientific Manuscript database

    The southern cattle fever tick, Rhipicephalus (Boophilus) microplus (Canestrini), is a haematophagous arachnid (Acari: Ixodidae) recognized globally as an economically important ectoparasite of cattle in tropical and subtropical agroecosystems. Populations of this invasive tick species around the wo...

  5. 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

  6. Saliva of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) inhibits classical and alternative complement pathways.

    PubMed

    Silva, Naylene C S; Vale, Vladimir F; Franco, Paula F; Gontijo, Nelder F; Valenzuela, Jesus G; Pereira, Marcos H; Sant'Anna, Mauricio R V; Rodrigues, Daniel S; Lima, Walter S; Fux, Blima; Araujo, Ricardo N

    2016-08-11

    Rhipicephalus (Boophilus) microplus is the main ectoparasite affecting livestock worldwide. For a successful parasitism, ticks need to evade several immune responses of their hosts, including the activation of the complement system. In spite of the importance of R. microplus, previous work only identified one salivary molecule that blocks the complement system. The current study describes complement inhibitory activities induced by R. microplus salivary components and mechanisms elicited by putative salivary proteins on both classical and alternative complement pathways. We found that R. microplus saliva from fully- and partially engorged females was able to inhibit both pathways. Saliva acts strongly at the initial steps of both complement activation pathways. In the classical pathway, the saliva blocked C4 cleavage, and hence, deposition of C4b on the activation surface, suggesting that the inhibition occurs at some point between C1q and C4. In the alternative pathway, saliva acts by binding to initial components of the cascade (C3b and properdin) thereby preventing the C3 convertase formation and reducing C3b production and deposition as well as cleavage of factor B. Saliva has no effect on formation or decay of the C6 to C8 components of the membrane attack complex. The saliva of R. microplus is able to inhibit the early steps of classical and alternative pathways of the complement system. Saliva acts by blocking C4 cleavage and deposition of C4b on the classical pathway activation surface and, in the alternative pathway, saliva bind to initial components of the cascade (C3b and properdin) thereby preventing the C3 convertase formation and the production and deposition of additional C3b.

  7. 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.

  8. [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.

  9. 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

    USDA-ARS?s Scientific Manuscript database

    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...

  10. Sequence characterization and immunogenicity of cystatins from the cattle tick Rhipicephalus (Boophilus) microplus.

    PubMed

    Parizi, Luís F; Githaka, Naftaly W; Acevedo, Carolina; Benavides, Uruguaysito; Seixas, Adriana; Logullo, Carlos; Konnai, Satoru; Ohashi, Kazuhiko; Masuda, Aoi; da Silva Vaz, Itabajara

    2013-12-01

    Various classes of endopeptidases and their inhibitors facilitate blood feeding and digestion in ticks. Cystatins, a family of tight-binding and reversible inhibitors of cysteine endopeptidases, have recently been found in several tick tissues. Moreover, vaccine trials using tick cystatins have been found to induce protective immune responses against tick infestation. However, the mode of action of tick cystatins is still poorly understood, limiting the elucidation of their physiological role. Against this background, we have investigated sequence characteristics and immunogenic properties of 5 putative cystatins from Rhipicephalus (Boophilus) microplus from Brazil and Uruguay. The similarity of the deduced amino acid sequences among cystatins from the Brazilian tick strain was 27-42%, all of which had a secretory signal peptide. The cystatin motif (QxVxG), a glycine in the N-terminal region, and the PW motif in the second hairpin loop in the C-terminal region are highly conserved in all 5 cystatins identified in this study. Four cysteine residues in the C terminus characteristic of type 2 cystatins are also present. qRT-PCR revealed differential expression patterns among the 5 cystatins identified, as well as variation in mRNA transcripts present in egg, larva, gut, salivary glands, ovary, and fat body tissues. One R. microplus cystatin showed 97-100% amino acid similarity between Brazilian and Uruguayan isolates. Furthermore, by in silico analysis, antigenic amino acid regions from R. microplus cystatins showed high degrees of homology (54-92%) among Rhipicephalus spp. cystatins. Three Brazilian R. microplus cystatins were expressed in Escherichia coli, and immunogenicity of the recombinant proteins were determined by vaccinating mice. Western blotting using mice sera indicated cross-reactivity between the cystatins, suggesting shared epitopes. The present characterization of Rhipicephalus spp. cystatins represents an empirical approach in an effort to evaluate

  11. 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.

  12. Characterization of pyrethroid resistance and susceptibility to coumaphos in Mexican Boophilus microplus (Acari: Ixodidae).

    PubMed

    Miller, R J; Davey, R B; George, J E

    1999-09-01

    Two patterns of pyrethroid resistance were characterized from Boophilus microplus (Canestrini) collected in Mexico. One was characteristic of a kdr mutation and the other involved esterase and cytochrome P450 enzyme systems. Very high resistance to permethrin, cypermethrin, and flumethrin, not synergized by TPP and PBO and high resistance to DDT, characterized the kdr-like pattern found in the Corrales and San Felipe strains. Esterase and cytochrome P450-dependent resistance was found in the Coatzacoalcos strain. It was characterized by resistance to permethrin, cypermethrin, and flumethrin, synergized by TPP and PBO, but no resistance to DDT. The Coatzacoalcos strain also showed 3.6-fold resistance to the organophosphate coumaphos. This factor appeared to be independent of pyrethroid resistance. Pyrethroid resistance patterns found in Mexico were similar to those found earlier in Australia. The significance of pyrethroid and coumaphos resistance to the U.S. cattle fever tick quarantine is discussed.

  13. Deltamethrin resistance in field populations of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in Jammu and Kashmir, India.

    PubMed

    Ahanger, R R; Godara, R; Katoch, R; Yadav, A; Bhutyal, A D S; Katoch, M; Singh, N K; Bader, M A

    2015-11-01

    Detection of resistance levels against deltamethrin in Rhipicephalus (Boophilus) microplus collected from six districts of Jammu and Kashmir (India) was carried out using the adult immersion test. The regression graphs of probit mortality of ticks plotted against log values of concentration of drug 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 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 six districts, resistance to deltamethrin at level I was detected in one district (RF = 1.9), at level II in two districts (RF = 7.08-10.07) and at level IV in three districts (RF = 96.08-288.72). The data generated on deltamethrin resistance status will help in formulating tick control strategy in the region.

  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. 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.

  18. 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. Crown Copyright 2009. Published by Elsevier Ltd. All rights reserved.

  19. 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.

  20. Identification of a dieldrin resistance-associated mutation in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Hope, Michelle; Menzies, Moira; Kemp, David

    2010-08-01

    The southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Acari: Ixodidae), is a major vector of tick fever organisms affecting cattle in many parts of the world, including Australia, Africa, and South America. Control of the southern cattle tick through acaricide use is an important approach in disease management. Resistance has emerged to many of the acaricides currently and previously used, including the cyclodienes. Although cyclodiene resistance mechanisms have been characterized in many insect species, this report is the first to identify mutations associated with dieldrin resistance in the cattle tick. A novel two base pair mutation in the GABA-gated chloride channel gene has been identified at position 868-9 and causes a codon change from threonine to leucine. Analysis of a small number of field-collected samples resistant to dieldrin shows this mutation has been maintained without selection pressure since the withdrawal of dieldrin in Australia > 20 yr ago. The mutation is not found in other laboratory-maintained strains of R. microplus that were subject to selection pressure with various acaricides.

  1. 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.

  2. 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

  3. 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. Copyright © 2013 Elsevier Inc. All rights reserved.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. Eprinomectin pour-on for control of Boophilus microplus (Canestrini) ticks (Acari: Ixodidae) on cattle.

    PubMed

    Aguirre, D H; Gaido, A B; Cafrune, M M; Castelli, M E; Mangold, A J; Guglielmone, A A

    2005-01-20

    The efficacy of a commercial pour-on formulation of eprinomectin, a macrocyclic lactone, against experimental infestations of Boophilus microplus (Canestrini) ticks was evaluated in two trials involving 27 Bos taurus calves. The first trial was designed to evaluate the effects of a single treatment at a dose of 0.5 mg/kg of body weight against standard size B. microplus females (4.5-8.0 mm long). A significant reduction in tick numbers (P<0.05, Wilcoxon test) was observed between treated calves as compared to untreated ones from Day 3 (44% efficacy) after treatment to the end of the trial on Day 28 (96.9%), with a peak efficacy of 97.1% on Day 21. In the second trial the effect of eprinomectin on standard size tick numbers, engorgement weight and fertility of female ticks from calves with a single treatment dose of 1 mg/kg on Day 0 and calves treated twice at a dose of 0.5 mg/kg on Days 0 and 4 was evaluated. An efficacy >93% was obtained from Day 2 to Day 28 after treatment in calves treated twice at 0.5 mg/kg, and to the end of the trial (Day 35) in calves treated once with 1 mg/kg. The 1mg/kg treatment provided >98% residual efficacy for at least 7 days. During the first part of the second trial the efficacy of eprinomectin resulted from a dramatic adverse effect on engorgement weight and fertility of female ticks, with 100% control on Day 5 (dosage of 1 mg/kg) and on Days 6 and 7 (two doses of 0.5 mg/kg). Following Day 7, most of the effect was due to reduction in the number of standard size female ticks.

  9. The embryogenesis of the tick Rhipicephalus (Boophilus) microplus: the establishment of a new chelicerate model system.

    PubMed

    Santos, Vitória Tobias; Ribeiro, Lupis; Fraga, Amanda; de Barros, Cíntia Monteiro; Campos, Eldo; Moraes, Jorge; Fontenele, Marcio Ribeiro; Araújo, Helena Marcolla; Feitosa, Natalia Martins; Logullo, Carlos; da Fonseca, Rodrigo Nunes

    2013-12-01

    Chelicerates, which include spiders, ticks, mites, scorpions, and horseshoe crabs, are members of the phylum Arthropoda. In recent years, several molecular experimental studies of chelicerates have examined the embryology of spiders; however, the embryology of other groups, such as ticks (Acari: Parasitiformes), has been largely neglected. Ticks and mites are believed to constitute a monophyletic group, the Acari. Due to their blood-sucking activities, ticks are also known to be vectors of several diseases. In this study, we analyzed the embryonic development of the cattle tick, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae). First, we developed an embryonic staging system consisting of 14 embryonic stages. Second, histological analysis and antibody staining unexpectedly revealed the presence of a population of tick cells with similar characteristics to the spider cumulus. Cumulus cell populations also exist in other chelicerates; these cells are responsible for the breaking of radial symmetry through bone morphogenetic protein signaling. Third, it was determined that the posterior (opisthosomal) embryonic region of R. microplus is segmented. Finally, we identified the presence of a transient ventral midline furrow and the formation and regression of a fourth leg pair; these features may be regarded as hallmarks of late tick embryogenesis. Importantly, most of the aforementioned features are absent from mite embryos, suggesting that mites and ticks do not constitute a monophyletic group or that mites have lost these features. Taken together, our findings provide fundamental common ground for improving knowledge regarding tick embryonic development, thereby facilitating the establishment of a new chelicerate model system. Copyright © 2013 Wiley Periodicals, Inc.

  10. Variation among Bm86 sequences in Rhipicephalus (Boophilus) microplus ticks collected from cattle across Thailand.

    PubMed

    Kaewmongkol, S; Kaewmongkol, G; Inthong, N; Lakkitjaroen, N; Sirinarumitr, T; Berry, C M; Jonsson, N N; Stich, R W; Jittapalapong, S

    2015-06-01

    Anti-tick vaccines based on recombinant homologues Bm86 and Bm95 have become a more cost-effective and sustainable alternative to chemical pesticides commonly used to control the cattle tick, Rhipicephalus (Boophilus) microplus. However, Bm86 polymorphism among geographically separate ticks is reportedly associated with reduced effectiveness of these vaccines. The purpose of this study was to investigate the variation of Bm86 among cattle ticks collected from Northern, Northeastern, Central and Southern areas across Thailand. Bm86 cDNA and deduced amino acid sequences representing 29 female tick midgut samples were 95.6-97.0 and 91.5-93.5 % identical to the nucleotide and amino acid reference sequences, respectively, of the Australian Yeerongpilly vaccine strain. Multiple sequence analyses of these Bm86 variants indicated geographical relationships and polymorphism among Thai cattle ticks. Two larger groups of cattle tick strains were discernable based on this phylogenetic analysis of Bm86, a Thai group and a Latin American group. Thai female and male cattle ticks (50 pairs) were also subjected to detailed morphological characterization to confirm their identity. The majority of female ticks had morphological features consistent with those described for R. (B.) microplus, whereas, curiously, the majority of male ticks were more consistent with the recently re-instated R. (B.) australis. A number of these ticks had features consistent with both species. Further investigations are warranted to test the efficacies of rBm86-based vaccines to homologous and heterologous challenge infestations with Thai tick strains and for in-depth study of the phylogeny of Thai cattle ticks.

  11. 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

  12. 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

  13. 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

  14. 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

  15. Molecular diagnosis of pyrethroid resistance in Mexican strains of Boophilus microplus (Acari: Ixodidae).

    PubMed

    Guerrero, Felix D; Li, Andrew Y; Hernandez, Ruben

    2002-09-01

    Polymerase chain reaction (PCR) diagnostic assays were used to identify possible resistance-associated roles of two amino acid substitutions found in pyrethroid resistance-associated genes of Boophilus microplus (Canestrini). Individual larvae from the San Felipe target site resistant strain and the Coatzacoalcos (Cz) metabolic resistant strain were separated into resistant and susceptible groups by larval packet bioassays and analyzed by PCR. A Phe --> Ile amino acid mutation in the sodium channel gene S6 transmembrane segment of domain III was found to have a close association with survival of acaricide treatments containing as high as 30% permethrin. As the permethrin dose was increased, an increase was seen in the proportion of surviving larvae that possessed two mutated sodium channel alleles. An Asp --> Asn amino acid substitution, originally found in high allele frequency in alleles of the CzEst9 esterase of the Cz strain, appeared to provide some resistance to permethrin. However, the presence of the mutation did not associate with resistance in the dose-response fashion seen with the sodium channel amino acid mutation. Resistance provided by CzEst9 might be more dependent on concentration of CzEst9 more so than the presence of a mutated allele.

  16. Perceptions of milk producers from Divinópolis, Minas Gerais, regarding Rhipicephalus (Boophilus) microplus control.

    PubMed

    da Rocha, Christiane Maria Barcellos Magalhães; Leite, Romário Cerqueira; Bruhn, Fábio Raphael Pascoti; Guimarães, Antônio Marcos; Furlong, John

    2011-01-01

    Semi-systematized interviews were conducted with 100 dairy cattle producers in the municipality of Divinópolis, Minas Gerais, with the aim of ascertaining their perceptions regarding the importance of Rhipicephalus (Boophilus) microplus and of combating it. Content analysis was performed and the frequency distribution of each of the variables was used to construct profiles of the producers interviewed. The production losses caused by ticks were perceived incompletely by the producers, who were unaware of the pathogen transmission caused by the parasite and the indirect losses through combating it, such as the cost of acaricide and labor. The combat operations were performed in a traditional manner, with an excessive number of inefficient treatments that aimed to control the level of infestation at that moment. The quality of the acaricide dipping/spraying applied was affected by the quality of the equipment used to apply the products, lack of knowledge of the mode of action of these products, lack of the specific information needed and lack of motivation caused by unawareness of the disadvantages of chemical combat. It was concluded that the lack of knowledge about combat methods and the acceptance of endemicity of the parasitosis were impediments to changing the realities encountered.

  17. 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.

  18. 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.

  19. 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).

  20. 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.

  1. Simulation of rotational grazing to evaluate integrated pest management strategies for Boophilus microplus (Acari: ixodidae) in Venezuela.

    PubMed

    Hernández-A, F; Teel, P D; Corson, M S; Grant, W E

    2000-09-20

    Ranchers in Venezuela historically have controlled the cattle-fever tick, Boophilus microplus (Canestrini), with acaricide treatments of cattle but no technical planning. We developed a simulation model to evaluate cattle-tick population dynamics in systematic pasture rotation systems and Integrated Pest Management (IPM) approaches to managing ticks in the tropical dry-forest ecological zone of Venezuela. Model output showed five generations of cattle-ticks produced each year throughout the dry and rainy seasons that occur in this zone. Sensitivity analyses showed disproportionately large changes in on-host B. microplus populations in response to small changes in larval mortality rates, such as those resulting from differences in the innate resistance of cattle to tick parasitism. Simulation results with 1-6 pasture systems suggest that adjusting the graze:rest sequence with systematic rotation among 4-6 pastures could suppress, but not eradicate, tick populations.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

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

    PubMed Central

    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. PMID:25174962

  7. Exploitation of chemical, herbal and nanoformulated acaricides to control the cattle tick, Rhipicephalus (Boophilus) microplus - A review.

    PubMed

    Banumathi, Balan; Vaseeharan, Baskaralingam; Rajasekar, Periyannan; Prabhu, Narayanan Marimuthu; Ramasamy, Palaniappan; Murugan, Kadarkarai; Canale, Angelo; Benelli, Giovanni

    2017-09-15

    The tick Rhipicephalus (Boophilus) microplus is a key vector of bacterial and protozoan diseases causing heavy economic losses directly and indirectly in animal husbandry. In the past decades, the control of ticks faced some major issues, such as the rapid development of resistance in targeted vectors and non-target effects on human health and the environment, due to the employ of synthetic acaricides and repellents. Eco-friendly pesticides for treating and controlling animal parasites such as ticks were mainly from medicinal plants and thus they form the richest entity for manufacturing resources for drugs. Even though there are efforts made to discover reliable plant-based acaricides to control ectoparasites in animal husbandry, the effective control of R. (B.) microplus ticks still represent a major challenge in current veterinary entomology. Recently, a wide number of promising attempts have been conducted to use herbal preparations and green-fabricated nanoparticles for the control of R. (B.) microplus. The aim of this review is to critically summarize and discuss the use of herbal preparations used in ethno-veterinary as well as green-fabricated nanoparticles as novel acaricides for the control of the cattle tick R. (B.) microplus. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  9. 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

    USDA-ARS?s Scientific Manuscript database

    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...

  10. A survey of fipronil- and ivermectin-resistant Rhipicephalus (Boophilus) microplus collected in Northern Mexico and the options for the management of acaricide-resistant ticks with pesticides

    USDA-ARS?s Scientific Manuscript database

    Six strains of Rhipicephalus (Boophilus) microplus collected from northern Mexico were surveyed for resistance to 5 classes of acaricide. All were resistant to permethrin. Two strains were resistant to amitraz and 3 were found to be resistant to coumaphos. Two strains were resistant to fipronil incl...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  12. Therapeutic and persistent efficacy of a long-acting (LA) formulation of ivermectin against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) and sera concentration through time in treated cattle

    USDA-ARS?s Scientific Manuscript database

    Concentration-time profile, therapeutic, and persistent efficacy of a single subcutaneous injection of cattle with a long-acting (LA) formulation of ivermectin at a concentration of 630 µg per kg of body weight was determined against Rhipicephalus (Boophilus) microplus. Ivermectin sera concentratio...

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

    USDA-ARS?s Scientific Manuscript database

    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,...

  14. Efficacy and blood sera analysis of a long-acting formulation of moxidectin against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae)on treated cattle

    USDA-ARS?s Scientific Manuscript database

    The therapeutic and persistent efficacy of a single subcutaneous injection of a long-acting (LA) formulation of moxidectin at a concentration of 1 mg per kg of body weight were determined against Rhipicephalus (Boophilus) microplus (Canestrini), along with the concentration-time blood sera profile i...

  15. Persistent efficacy and blood sera analysis of a long-acting (LA) formulation of moxidectin against Rhipicephalus (Boophilus) microplus on treated cattle

    USDA-ARS?s Scientific Manuscript database

    The therapeutic and persistent efficacy of a single subcutaneous injection of a long-acting (LA) formulation of moxidectin at a concentration of 1 mg per kg of body weight were determined against Rhipicephalus (Boophilus) microplus (Canestrini), along with the concentration-time blood sera profile i...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  17. 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

    USDA-ARS?s Scientific Manuscript database

    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...

  18. Efficacy of amitraz applied as a dip against an amitraz-resistant strain of Rhipicephalus (Boophilus) microplus (Acari:Ixodidae) infested on cattle

    USDA-ARS?s Scientific Manuscript database

    Selection pressure with amitraz applied to Rhipicephalus (Boophilus) microplus (Canestrini) in 13 out of 18 generations resulted in a 28.9-fold increase in resistance level, indicating a shift in the phenotypic composition of the ticks from a heterogenous mixture of both susceptible and moderately r...

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. Toxicity of Piper aduncum L. (Piperales: Piperaceae) from the Amazon forest for the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Silva, Wilson Castro; Martins, João Ricardo de Souza; de Souza, Hellen Emília Menezes; Heinzen, Horacio; Cesio, Maria Verônica; Mato, Mauricio; Albrecht, Francine; de Azevedo, João Lúcio; de Barros, Neiva Monteiro

    2009-10-14

    The mortality of 14-21-day-old Rhipicephalus (Boophilus) microplus larvae, and the mortality and fertility of groups of engorged adult females exposed to different concentrations of hexane, ethyl acetate and ethanol extracts of spiked pepper (Piper aduncum) were evaluated, using a completely randomized design with five treatment groups, two control groups, and two replicates for the larvae and five replicates for the adult females. Similar methodology was used to investigate the toxicity of the essential oil hydro-distillate (94.84% dillapiole) obtained from the P. aduncum crude hexane extract. The LC(50) of the hexane extract was 9.30 mg ml(-1) for larvae and the reproduction reduction ranged from 12.48% to 54.22%, while 0.1mg/ml(-1) of the essential oil induced 100% mortality in larvae. Literature reports on natural products active against R. microplus were listed and compared with the results presented here. These results indicate that P. aduncum extracts, and particularly its essential oil, are potential alternative control agents for R. microplus.

  6. Acaricidal action of destruxins produced by a marine-derived Beauveria felina on the bovine tick Rhipicephalus (Boophilus) microplus.

    PubMed

    Morais-Urano, Raquel P; Chagas, Ana C S; Berlinck, Roberto G S

    2012-11-01

    The increasing resistance of Rhipicephalus (Boophilus) microplus tick to commercial insecticides requires alternative methods for the control of this cattle plague. The enthomopathogenic fungus Beauveria felina produces destruxins in culture media, cyclic depsipeptides which display an array of biological activities. The present investigation aimed to evaluate the acaricide action of destruxins isolated from B. felina culture media on R. (B.) microplus engorged females. B. felina was grown in MF medium under 19 different growth conditions. HPLC-PDA analysis of chromatographic fractions obtained from the 19 different growth media extracts indicated the presence of destruxins in all lipophylic fractions. Such fractions were combined and subjected to separation by HPLC. Fractions containing distinct destruxins composition were tested against R. (B.) microplus. Two fractions, composed of destruxin Ed(1) and pseudodestruxin B and/or pseudodestruxin C (fraction P1) as well as by hydroxyhomodestruxin B and/or destruxin D and/or roseotoxin C (fraction P7), displayed 30% and 28.7% acaricidal efficacy, respectively. This activity profile in such low concentration is adequate to consider destruxins as potential leading compounds to be developed for tick biological control. Copyright © 2012 Elsevier Inc. All rights reserved.

  7. 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.

  8. 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

  9. 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.

  10. Identification and characterization of Rhipicephalus (Boophilus) microplus candidate protective antigens for the control of cattle tick infestations.

    PubMed

    Almazán, Consuelo; Lagunes, Rodolfo; Villar, Margarita; Canales, Mario; Rosario-Cruz, Rodrigo; Jongejan, Frans; de la Fuente, José

    2010-01-01

    The cattle ticks, Rhipicephalus (Boophilus) spp., affect cattle production in tropical and subtropical regions of the world. Tick vaccines constitute a cost-effective and environmentally friendly alternative to tick control. The recombinant Rhipicephalus microplus Bm86 antigen has been shown to protect cattle against tick infestations. However, variable efficacy of Bm86-based vaccines against geographic tick strains has encouraged the research for additional tick-protective antigens. Herein, we describe the analysis of R. microplus glutathione-S transferase, ubiquitin (UBQ), selenoprotein W, elongation factor-1 alpha, and subolesin (SUB) complementary DNAs (cDNAs) by RNA interference (RNAi) in R. microplus and Rhipicephalus annulatus. Candidate protective antigens were selected for vaccination experiments based on the effect of gene knockdown on tick mortality, feeding, and fertility. Two cDNA clones encoding for UBQ and SUB were used for cattle vaccination and infestation with R. microplus and R. annulatus. Control groups were immunized with recombinant Bm86 or adjuvant/saline. The highest vaccine efficacy for the control of tick infestations was obtained for Bm86. Although with low immunogenic response, the results with the SUB vaccine encourage further investigations on the use of recombinant subolesin alone or in combination with other antigens for the control of cattle tick infestations. The UBQ peptide showed low immunogenicity, and the results of the vaccination trial were inconclusive to assess the protective efficacy of this antigen. These experiments showed that RNAi could be used for the selection of candidate tick-protective antigens. However, vaccination trials are necessary to evaluate the effect of recombinant antigens in the control of tick infestations, a process that requires efficient recombinant protein production and formulation systems.

  11. 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

  12. Use of an allele-specific polymerase chain reaction assay to genotype pyrethroid resistant strains of Boophilus microplus (Acari: Ixodidae).

    PubMed

    Guerrero, F D; Davey, R B; Miller, R J

    2001-01-01

    A polymerase chain reaction-based assay was developed to detect the presence of a pyrethroid resistance-associated amino acid substitution in Boophilus microplus (Canestrini). The assay uses a simple method for the extraction of genomic DNA from individual larvae and genotypes individuals for the presence of a Phe-->Ile amino acid substitution in the S6 transmembrane segment of domain III of the para-like sodium channel, clearly distinguishing heterozygotes from homozygotes. High frequencies for this amino acid substitution were found in the Corrales and San Felipe strains, which have target site insensitivity mechanisms for pyrethroid resistance. The Caporal resistant strain contained lower yet substantial numbers of amino acid-substituted alleles. Low amino acid substitution frequencies were found in the susceptible reference Gonzales strain and the Coatzacoalcos strain, which has metabolic esterase-mediated pyrethroid resistance. The amino acid substitution was not found in six other strains that were susceptible to pyrethroids.

  13. 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

  14. Reduction in pathogenicity of Babesia bovis for its tick vector, Boophilus microplus, after rapid blood passage in splenectomized calves.

    PubMed

    Dalgliesh, R J; Stewart, N P; Duncalfe, F

    1981-01-01

    Two strains of Babesia bovis that had been serially blood passaged in splenectomized calves 27 to 33 times, a procedure known to have reduced their virulence for normal cattle, were shown to have low pathogenicity for replete, female Boophilus microplus. In comparison with a strain of B. bovis unmodified by repeated blood passage, the two modified strains infected higher proportions of ticks and produced comparable numbers of morphologically similar parasites in their haemolymph, but killed significantly fewer of them. Red discolouration of haemolymph was observed in many ticks infected with the unmodified strain, but in none of those infected with the modified strains. It is suggested that the modified strains have lost a quality causing pathological effects on the gut cells of infected ticks.

  15. 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.

  16. Bovine immunoprotection against Rhipicephalus (Boophilus) microplus with recombinant Bm86-Campo Grande antigen

    USDA-ARS?s Scientific Manuscript database

    The southern cattle fever tick, Rhipicephalus microplus, is arguably the most economically important external parasite of cattle globally. The inappropriate use of readily available chemical acaricides has driven the evolution of resistance in populations of R. microplus. Anti-tick vaccines represen...

  17. Ixodid ticks of traditionally managed cattle in central Nigeria: where Rhipicephalus (Boophilus) microplus does not dare (yet?)

    PubMed Central

    2013-01-01

    Background Ticks and tick-borne diseases (TBDs) undermine cattle fitness and productivity in the whole of sub-Saharan Africa, including Nigeria. The aim of this study was to document the composition of tick species, assessing the burden of infestation, in traditionally managed cattle in an area of central Nigeria where acaricides have not been used historically. Methods The study was carried out in September 2010 in 9 villages belonging to three neighbouring local government areas in Plateau State, Nigeria. In each village all visible adult ticks were collected from at least 15 cattle (mean number = 25). Collected ticks were preserved in 70% ethanol to be counted and morphologically identified to the species level. Results A total of 5011 ixodid ticks (1935 males and 3076 females) were collected from 228 cattle, comprising 14 calves, 33 juveniles, and 181 adults. Three tick genera (i.e., Amblyomma, Hyalomma, and Rhipicephalus, including the Boophilus sub-genus) and 11 species were identified. The most prevalent species was Rhipicephalus (Boophilus) decoloratus (41.4%), followed by Rhipicephalus (Boophilus) annulatus (15.4%), Rhipicephalus guilhoni (12.0%), Rhipicephalus (Boophilus) geigyi (7.6%), Hyalomma truncatum (7.4%), Amblyomma variegatum (6.3%), Rhipicephalus simus Group (4.0%), Rhipicephalus turanicus (1.2%), Rhipicephalus sanguineus (0.3%), Hyalomma rufipes (0.2%), and Rhipicephalus lunulatus (n = 1). Mean tick loads recorded were relatively high (22 ± 1.4), in spite of the practice of hand removal of ticks traditionally undertaken by the Fulani pastoralists in the area. Calves bore a significantly lower tick burden than adults (p = 0.004). Rhipicephalus (Boophilus) microplus was not found in the area, suggesting that the eastbound expansion of this tick species in West Africa, has not yet reached central Nigeria. Conclusions This study ascertained the presence of a broad variety of cattle tick species, most of which are of veterinary

  18. Ixodid ticks of traditionally managed cattle in central Nigeria: where Rhipicephalus (Boophilus) microplus does not dare (yet?).

    PubMed

    Lorusso, Vincenzo; Picozzi, Kim; de Bronsvoort, Barend M C; Majekodunmi, Ayodele; Dongkum, Charles; Balak, Gyang; Igweh, Augustine; Welburn, Susan C

    2013-06-07

    Ticks and tick-borne diseases (TBDs) undermine cattle fitness and productivity in the whole of sub-Saharan Africa, including Nigeria. The aim of this study was to document the composition of tick species, assessing the burden of infestation, in traditionally managed cattle in an area of central Nigeria where acaricides have not been used historically. The study was carried out in September 2010 in 9 villages belonging to three neighbouring local government areas in Plateau State, Nigeria. In each village all visible adult ticks were collected from at least 15 cattle (mean number = 25). Collected ticks were preserved in 70% ethanol to be counted and morphologically identified to the species level. A total of 5011 ixodid ticks (1935 males and 3076 females) were collected from 228 cattle, comprising 14 calves, 33 juveniles, and 181 adults. Three tick genera (i.e., Amblyomma, Hyalomma, and Rhipicephalus, including the Boophilus sub-genus) and 11 species were identified. The most prevalent species was Rhipicephalus (Boophilus) decoloratus (41.4%), followed by Rhipicephalus (Boophilus) annulatus (15.4%), Rhipicephalus guilhoni (12.0%), Rhipicephalus (Boophilus) geigyi (7.6%), Hyalomma truncatum (7.4%), Amblyomma variegatum (6.3%), Rhipicephalus simus Group (4.0%), Rhipicephalus turanicus (1.2%), Rhipicephalus sanguineus (0.3%), Hyalomma rufipes (0.2%), and Rhipicephalus lunulatus (n = 1). Mean tick loads recorded were relatively high (22 ± 1.4), in spite of the practice of hand removal of ticks traditionally undertaken by the Fulani pastoralists in the area. Calves bore a significantly lower tick burden than adults (p = 0.004). Rhipicephalus (Boophilus) microplus was not found in the area, suggesting that the eastbound expansion of this tick species in West Africa, has not yet reached central Nigeria. This study ascertained the presence of a broad variety of cattle tick species, most of which are of veterinary importance. The presence of each tick

  19. Sequence polymorphism in acetylcholinesterase transcripts and genotyping survey of BmAChE1 in laboratory and Mexican strains of Rhipicephalus (Boophilus) microplus

    USDA-ARS?s Scientific Manuscript database

    BmAChE1, BmAChE2, and BmAChE3 cDNAs of Rhipicephalus (Boophilus) microplus were sequenced and found to exhibit significant polymorphism. A portion of the predicted amino acid substitutions in BmAChE1, BmAChE2 and BmAChE3 were found predominantly in organophosphate-resistant (OP-R) strains, but most ...

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

    PubMed

    Zivkovic, Zorica; Esteves, Eliane; Almazán, Consuelo; Daffre, Sirlei; Nijhof, Ard M; Kocan, Katherine M; Jongejan, Frans; de la Fuente, José

    2010-03-18

    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. 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) significantly reduced

  1. 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

  2. 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

  3. 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. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. 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.

  5. 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.

  6. Immune responses against recombinant tick antigen, Bm95, for the control of Rhipicephalus (Boophilus) microplus ticks in cattle.

    PubMed

    Kumar, Amit; Garg, Rajat; Yadav, C L; Vatsya, Stuti; Kumar, R R; Sugumar, Parthasarthy; Chandran, Dev; Mangamoorib, Lakshmi Narasu; Bedarkar, S N

    2009-10-28

    Immune responses against Bm95 recombinant cattle tick antigen and its protective efficacy for control of Rhipicephalus (Boophilus) microplus ticks were determined in experimental crossbred cow calves. Anti-Bm95 antibody titers, as assessed by indirect ELISA, in immunized calves ranged from 196.1+/-13.7 on day 0 to 7979.9+/-312.5 on day 110 post-primary immunization. The rise in antibody titer was statistically significant (p<0.01) throughout the study period. Besides this, constantly higher lymphoproliferative response (LPR), as assessed by lymphocyte stimulation test, was observed from 10 days post-immunization, but a positive LPR of antigen stimulated cells in immunized animals was recorded only on day 50 and day 70 post-immunization. Following challenge of immunized calves with larvae of R. microplus, significant increase (p<0.01) in rejection percentage, mean number of damaged ticks, mean percentage of dead ticks, and decrease in engorgement weight were recorded in immunized animals. Also, there were significant differences (p<0.01) in preoviposition period, oviposition period, egg mass weight and percent hatchability between the immunized and control calves. The percent reduction in number of adult females in vaccinated calves, reduction in mean weight of egg masses, percent reduction in mean weight and reduction in fertility of engorged females collected from vaccinated calves were determined and the efficacy of Bm95 recombinant cattle tick antigen was 81.27%.

  7. [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.

  8. 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.

  9. 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.

  10. Review of cattle ticks (Acari, Ixodida) in Ivory Coast and geographic distribution of Rhipicephalus (Boophilus) microplus, an emerging tick in West Africa.

    PubMed

    Boka, O M; Achi, L; Adakal, H; Azokou, A; Yao, P; Yapi, Y G; Kone, M; Dagnogo, K; Kaboret, Y Y

    2017-04-01

    The exotic tick Rhipicephalus (Boophilus) microplus was discovered in Ivory Coast in 2007 and then gradually in other countries in West Africa. It is known to induce significant losses in farming and to replace other species of the same genus. In order to contribute to improve health and productivity of cattle in Ivory Coast regarding the emergence of this dreaded tick, a study was conducted to determine the current geographic distribution of the tick R. (B.) microplus and review cattle ticks in general. To this end, 23,460 ticks were collected from 180 farms located throughout the country. Ten species of ticks belonging to the genus Rhipicephalus (including those of the subgenus Boophilus), Hyalomma and Ambyomma were identified. It was found that the exotic tick R. (B.) microplus has invaded the entire Ivorian territory and is now the main cattle tick (63.6% of ticks collected), followed by Amblyomma variegatum that remains still dominant in the North. The population of indigenous species of Rhipicephalus (Boophilus) is in drastic decline.

  11. Local immune response against larvae of Rhipicephalus (Boophilus) microplus in Bos taurus indicus and Bos taurus taurus cattle.

    PubMed

    Constantinoiu, C C; Jackson, L A; Jorgensen, W K; Lew-Tabor, A E; Piper, E K; Mayer, D G; Venus, B; Jonsson, N N

    2010-06-01

    Bos taurus indicus cattle are less susceptible to infestation with Rhipicephalus (Boophilus) microplus than Bos taurus taurus cattle but the immunological basis of this difference is not understood. We compared the dynamics of leukocyte infiltrations (T cell subsets, B cells, major histocompatibility complex (MHC) class II-expressing cells, granulocytes) in the skin near the mouthparts of larvae of R. microplus in B. t. indicus and B. t. taurus cattle. Previously naïve cattle were infested with 50,000 larvae (B. t. indicus) or 10,000 larvae (B. t. taurus) weekly for 6 weeks. One week after the last infestation all of the animals were infested with 20,000 larvae of R. microplus. Skin punch biopsies were taken from all animals on the day before the primary infestation and from sites of larval attachment on the day after the first, second, fourth and final infestations. Infiltrations with CD3(+), CD4(+), CD8(+) and gammadelta T cells followed the same pattern in both breeds, showing relatively little change during the first four weekly infestations, followed by substantial increases at 7 weeks post-primary infestation. There was a tendency for more of all cell types except granulocytes to be observed in the skin of B. t. indicus cattle but the differences between the two breeds were consistently significant only for gammadelta T cells. Granulocyte infiltrations increased more rapidly from the day after infestation and were higher in B. t. taurus cattle than in B. t. indicus. Granulocytes and MHC class II-expressing cells infiltrated the areas closest to the mouthparts of larvae. A large volume of granulocyte antigens was seen in the gut of attached, feeding larvae. (c) 2010. Published by Elsevier Ltd. All rights reserved.

  12. 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.

  13. Antimicrobial activity in the tick Rhipicephalus (Boophilus) microplus eggs: Cellular localization and temporal expression of microplusin during oogenesis and embryogenesis.

    PubMed

    Esteves, E; Fogaça, A C; Maldonado, R; Silva, F D; Manso, P P A; Pelajo-Machado, M; Valle, D; Daffre, S

    2009-08-01

    Arthropods display different mechanisms to protect themselves against infections, among which antimicrobial peptides (AMPs) play an important role, acting directly against invader pathogens. We have detected several factors with inhibitory activity against Candida albicans and Micrococcus luteus on the surface and in homogenate of eggs of the tick Rhipicephalus (Boophilus) microplus. One of the anti-M. luteus factors of the egg homogenate was isolated to homogeneity. Analysis by electrospray mass spectrometry (ESI-MS) revealed that it corresponds to microplusin, an AMP previously isolated from the cell-free hemolymph of R. (B.) microplus. Reverse transcription (RT) quantitative polymerase chain reactions (qPCR) showed that the levels of microplusin mRNA gradually increase along ovary development, reaching an impressive highest value three days after the adult females have dropped from the calf and start oviposition. Interestingly, the level of microplusin mRNA is very low in recently laid eggs. An enhance of microplusin gene expression in eggs is observed only nine days after the onset of oviposition, achieving the highest level just before the larva hatching, when the level of expression decreases once again. Fluorescence microscopy analysis using an anti-microplusin serum revealed that microplusin is present among yolk granules of oocytes as well as in the connecting tube of ovaries. These results, together to our previous data, suggest that microplusin may be involved not only in protection of adult female hemocele, but also in protection of the female reproductive tract and embryos, what points this AMP as a considerable target for development of new methods to control R. (B.) microplus as well as the vector-borne pathogens.

  14. 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. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. 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.

  16. 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.

  17. 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. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. 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. Copyright © 2017 Elsevier

  19. Biochemical characterization of a Kunitz type inhibitor similar to dendrotoxins produced by Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) hemocytes.

    PubMed

    Lima, Cássia A; Torquato, Ricardo J S; Sasaki, Sergio D; Justo, Giselle Z; Tanaka, Aparecida S

    2010-02-10

    A novel chymotrypsin inhibitor identified in fat body and hemocyte cDNA libraries of Boophilus microplus was named BmCI (B. microplus Chymotrypsin Inhibitor) (Genbank EU636772). The putative BmCI amino acid sequence presented a 22-residue-signal peptide and 58-residue-mature protein. BmCI amino acid sequence analysis allowed its classification as a Kunitz-BPTI inhibitor with six cysteine residues, a theoretical pI of 7.8, and the presence of Tyr at P1 position in the putative reactive site, suggesting inhibitory activity toward chymotrypsin. In this work, we reported the biochemical characterization of BmCI. The recombinant BmCI expressed in yeast Pichia pastoris was purified by size exclusion and reverse phase chromatographies. rBmCI expression yield was of 1mgL(-1) of culture. Purified rBmCI confirmed its chymotrypsin inhibitory activity with a low K(i) (6.2pM). The BmCI gene expression analysis by semi-quantitative RT-PCR indicated its transcription in the hemocytes, salivary gland and ovary. The cytotoxic activity of purified rBmCI was demonstrated in BALB/c 3T3 mouse fibroblasts. As assessed by the MTT reduction assay, rBMCI induced a dose-dependent decrease in 3T3 fibroblasts viability (IC(50)=8microM). Moreover, flow cytometry analysis revealed that rBmCI is able to induce apoptosis, whereas no effect was observed on cell cycle progression. In conclusion, we demonstrated that rBmCI is cytotoxic against mammalian cells and obtained evidence that this growth inhibition is caused by an apoptosis-inducing activity.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  1. 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

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

    USDA-ARS?s Scientific Manuscript database

    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...

  3. Acaricide resistance of Rhipicephalus (Boophilus) microplus in State of Mato Grosso do Sul, Brazil

    USDA-ARS?s Scientific Manuscript database

    This study was performed to obtain an emerging view of the epidemiology of acaricide resistance in populations of R. microplus from Mato Grosso do Sul state in Brazil. Twenty four tick samples were collected from cities in the state where farmers reported concerns about resistance or failure of tick...

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

    USDA-ARS?s Scientific Manuscript database

    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. ...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

  7. 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

  8. 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.

  9. 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.

  10. 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

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. Efficacy of fipronil (1.0 mg/kg) against Rhipicephalus (Boophilus) microplus strains resistant to ivermectin (0.63 mg/kg).

    PubMed

    Lopes, Welber Daniel Zanetti; Cruz, Breno Cayeiro; Teixeira, Weslen Fabrício Pires; Felippelli, Gustavo; Maciel, Willian Giquelin; Buzzulini, Carolina; Gomes, Lucas V Costa; Favero, Flávia; Soares, Vando Edésio; Bichuette, Murilo A; de Oliveira, Gilson Pereira; da Costa, Alvimar José

    2014-08-01

    The present study aimed to evaluate the efficacy of fipronil (1 mg/kg) against three strains of ivermectin-resistant Rhipicephalus (Boophilus) microplus (R. (B.) microplus), naturally infesting cattle from different states of Brazil. Three rural properties with a history of macrocyclic lactones ineffectiveness against the cattle tick, and low frequency use of fipronil in the herd, were selected for the study. The animals were randomized according to the mean tick counts, performed on days -3, -2 and -1, into three groups with 10 animals each: T01, control (saline solution); T02, subcutaneous ivermectin (0.63 mg/kg) and T03, topical fipronil (1 mg/kg). Treatment was performed on day 0. Counts of partially engorged female ticks were performed on days 3, 7 and 14 post-treatment (DPT), and then every 7 days until the 49th DPT. In all three experiments, it was possible to diagnose resistance of R. (B.) microplus to ivermectin (0.63 mg/kg). The maximum efficacy (arithmetic mean) obtained for ivermectin was 64% in experiment II. On the other hand, the formulation containing fipronil (1 mg/kg) reached high efficacy values (≥97%) in all three experiments. The results from all experiments in this study demonstrate the high efficacy of 1mg/kg fipronil, administered pour-on in naturally infested cattle, against strains of R. (B.) microplus that are resistant to 630 mcg/kg ivermectin. Copyright © 2014 Elsevier B.V. All rights reserved.

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

    PubMed Central

    2011-01-01

    Background 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) microplus, causes significant economic losses in cattle production worldwide and this drives us to further understand their biology so that effective control measures can be developed. To be able to provide new insights into the biology of cattle ticks and to expand the repertoire of tick miRNAs we utilized Illumina technology to sequence the small RNA transcriptomes derived from various life stages and selected organs of R. microplus. Results To discover and profile cattle tick miRNAs we employed two complementary approaches, one aiming to find evolutionary conserved miRNAs and another focused on the discovery of novel cattle-tick specific miRNAs. We found 51 evolutionary conserved R. microplus miRNA loci, with 36 of these previously found in the tick Ixodes scapularis. The majority of the R. microplus miRNAs are perfectly conserved throughout evolution with 11, 5 and 15 of these conserved since the Nephrozoan (640 MYA), Protostomian (620MYA) and Arthropoda (540 MYA) ancestor, respectively. We then employed a de novo computational screening for novel tick miRNAs using the draft genome of I. scapularis and genomic contigs of R. microplus as templates. This identified 36 novel R. microplus miRNA loci of which 12 were conserved in I. scapularis. Overall we found 87 R. microplus miRNA loci, of these 15 showed the expression of both miRNA and miRNA* sequences. R. microplus miRNAs showed a variety of expression profiles, with the evolutionary-conserved miRNAs mainly expressed in all life stages at various levels, while the expression of novel tick-specific miRNAs was mostly limited to particular life stages and/or tick organs. Conclusions Anciently acquired mi

  19. 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

  20. 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

  1. In Vitro Detection of Acaricidal Resistance Status of Rhipicephalus (Boophilus) microplus against Commercial Preparation of Deltamethrin, Flumethrin, and Fipronil from North Gujarat, India

    PubMed Central

    Shyma, K. P.; Gupta, Jay Prakash; Singh, Veer; Patel, K. K.

    2015-01-01

    Rhipicephalus (Boophilus) microplus is the most common tick species in India infesting cattle and buffaloes and causing significant economic losses to dairy and leather industries by adversely affecting the milk production and quality of hides. A study to evaluate the acaricide resistance status of Rhipicephalus (Boophilus) microplus to deltamethrin, flumethrin, and fipronil was conducted on the samples collected from organized and unorganized farms of North Gujarat state, where treatment failures were reported frequently. Adult Immersion Test (AIT) and Larval Packet Test (LPT) were conducted using field strain for determination of 50 and 95% lethal concentration of deltamethrin, flumethrin, and fipronil. Results obtained by the Adult Immersion Test showed low grade resistance (level I, RF > 5) has been developed against both deltamethrin and fipronil. However, deltamethrin by performing Larval Packet Test showed moderate grade resistance (level II, RF > 25). Larval packet performed by flumethrin also revealed low grade resistance, level I. The data on field status of acaricide resistance from the area with diversified animal genetic resources will be helpful to adopt suitable strategy to overcome the process of development of resistance in ticks. PMID:26788362

  2. 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.

  3. 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

  4. Exopolyphosphatases in nuclear and mitochondrial fractions during embryogenesis of the hard tick Rhipicephalus (Boophilus) microplus.

    PubMed

    Campos, Eldo; Façanha, Arnoldo R; Costa, Evenilton P; da Silva Vaz, Itabajara; Masuda, Aoi; Logullo, Carlos

    2008-11-01

    The present work evaluated polyphosphate (poly P) metabolism in nuclear and mitochondrial fractions during Rhipicephalus microplus embryogenesis. Nuclear poly P decreased and activity of exopolyphosphatase (PPX - polyphosphate-phosphohydrolases; EC 3.6.1.11) increased after embryo cellularization until the end of embryogenesis. The utilization of mitochondrial poly P content occurred between embryo cellularization and segmentation stages. Increasing amounts of total RNA extracted from eggs progressively enhanced nuclear PPX activity, whereas it exerted no effect on mitochondrial PPX activity. The decline in total poly P content after the 7th day of embryogenesis does not reflect the free P(i) increase and the total poly P chain length decrease after embryo cellularization. The Km(app) utilizing poly P(3), poly P(15) and poly P(65) as substrate was almost the same for the nuclear fraction (around 1muM), while the affinity for substrate in mitochondrial fraction was around 10 times higher for poly P(3) (Km(app) = 0.2muM) than for poly P(15) (Km(app) = 2.8muM) and poly P(65) (Km(app) = 3.6muM). PPX activity was stimulated by a factor of two by Mg2+ and Co2+ in the nuclear fraction and only by Mg2+ in the mitochondrial fraction. Heparin (20microg/mL) inhibited nuclear and mitochondrial PPX activity in about 90 and 95% respectively. Together, these data are consistent with the existence of two different PPX isoforms operating in the nuclei and mitochondria of the hard tick R. microplus with distinct metal dependence, inhibitor and activator sensitivities. The data also shed new light on poly P biochemistry during arthropod embryogenesis, opening new routes for future comparative studies on the physiological roles of different poly P pools distributed over cell compartments.

  5. [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.

  6. 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.

  7. 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.

  8. Selection of reference genes for quantitative RT-PCR studies in Rhipicephalus (Boophilus) microplus and Rhipicephalus appendiculatus ticks and determination of the expression profile of Bm86

    PubMed Central

    2009-01-01

    Background For accurate and reliable gene expression analysis, normalization of gene expression data against reference genes is essential. In most studies on ticks where (semi-)quantitative RT-PCR is employed, normalization occurs with a single reference gene, usually β-actin, without validation of its presumed expression stability. The first goal of this study was to evaluate the expression stability of commonly used reference genes in Rhipicephalus appendiculatus and Rhipicephalus (Boophilus) microplus ticks. To demonstrate the usefulness of these results, an unresolved issue in tick vaccine development was examined. Commercial vaccines against R. microplus were developed based on the recombinant antigen Bm86, but despite a high degree of sequence homology, these vaccines are not effective against R. appendiculatus. In fact, Bm86-based vaccines give better protection against some tick species with lower Bm86 sequence homology. One possible explanation is the variation in Bm86 expression levels between R. microplus and R. appendiculatus. The most stable reference genes were therefore used for normalization of the Bm86 expression profile in all life stages of both species to examine whether antigen abundance plays a role in Bm86 vaccine susceptibility. Results The transcription levels of nine potential reference genes: β-actin (ACTB), β-tubulin (BTUB), elongation factor 1α (ELF1A), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), glutathione S-transferase (GST), H3 histone family 3A (H3F3A), cyclophilin (PPIA), ribosomal protein L4 (RPL4) and TATA box binding protein (TBP) were measured in all life stages of R. microplus and R. appendiculatus. ELF1A was found to be the most stable expressed gene in both species following analysis by both geNorm and Normfinder software applications, GST showed the least stability. The expression profile of Bm86 in R. appendiculatus and R. microplus revealed a more continuous Bm86 antigen abundance in R. microplus throughout its

  9. 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

  10. Conditions for stable parapatric coexistence between Boophilus decoloratus and B. microplus ticks: a simulation study using the competitive Lotka-Volterra model.

    PubMed

    Zeman, Petr; Lynen, Godelieve

    2010-12-01

    The autochthonous tick Boophilus decoloratus, and the invasive species Bo. microplus, the tick most threatening the livestock industry in Africa, show complex interactions in their interspecific rivalry. This study was conducted to specify the conditions under which the two competitors can co-exist in equilibrium, and to provide insight into their climate-dependant parapatric distribution in Tanzania. A model of the Lotka-Volterra type was used, taking into account population dispersal and interactions of various kinds. If the model allowed for immunity-mediated competition on cattle, reproductive interference, and an external mortality factor, it explained fairly well the field observation that the borderline between these ticks loosely follows the 22-23°C isotherm and the 58 mm isohyet (i.e. ~700 mm of annual rainfall total). Simulations fully compatible with the pattern of real co-existing populations of Bo. decoloratus and Bo. microplus, characterized by a pronounced population density trough and mutual exclusion of the two ticks on cattle in an intermediary zone between their distributional ranges, were, however, achieved only if the model also implemented a hypothetical factor responsible for some mortality upon encounter of one tick with the other, interpretable as an interaction through a shared pathogen(s). This study also demonstrated the importance of non-cattle hosts, enabling the autochthon to avoid competition with Bo. microplus, for the behaviour of the modelled system. The simulations indicate that a substantial reduction of wildlife habitats and consequently of Bo. decoloratus refugia, may accelerate the replacement of Bo. decoloratus with Bo. microplus much faster than climatic changes might do.

  11. 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

  12. Factors related to cattle infestation level and resistance to acaricides in Boophilus microplus tick populations in New Caledonia.

    PubMed

    Bianchi, M W; Barré, N; Messad, S

    2003-02-28

    Boophilus microplus, even in the absence of babesiosis, is a major disease of the cattle in New Caledonia where the particular farming system associates continental European breeds and a tropical climate tempered by the Pacific Ocean. In order to have a better understanding of the factors involved in cattle tick infestation, to decrease the possible wastage and use of chemicals and to increase the lifetime of the acaricides, the veterinary authorities investigated the conditions of the chemical treatments. A survey among 148 cattle farms of the whole of New Caledonia was carried out and factors that explain the development of tick resistance and cattle infestation have been determined. From this survey, three typologies for the main characteristics of the farms have been set up, the technical practices of the farmers and the tick control measures applied by the farmers, respectively. Some variables are significantly associated with the tick resistance to deltamethrin but their contribution to the explanation model is always moderate: farms in the south, with a positive resistance gradient from east to west, absence of bush fire and membership to a cattle farmers organization. The more the farmers have intensified their breeding-male castration, weaning, heifer separation, drenching, etc.-and pasture-high stock rate, mowing, extra feeding of the cows, many paddocks, etc.-techniques, the higher was the probability for the ticks in their farm to be resistant to deltamethrin. The technical details of the acaricide treatment had a low contribution to the explanation model. However, the use of a spray generated more resistance than a dip. Furthermore, there is a negative resistance gradient when the farmers increased the treatment interval average. Considering infestation, none of the variables from the three typologies were associated with the two infestation variables (1: semi-engorged tick females and 2: other ticks) at the herd level. However, the seven studied

  13. 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

  14. 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.

  15. 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

  16. 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

  17. Comparative in vitro anti-tick efficacy of commercially available products and newly developed phyto-formulations against field collected and resistant tick lines of Rhipicephalus (Boophilus) microplus.

    PubMed

    Ajith Kumar, K G; Sharma, Anil Kumar; Kumar, Sachin; Ray, D D; Rawat, A K S; Srivastava, Sharad; Ghosh, Srikant

    2016-12-01

    Rhipicephalus (Boophilus) microplus is considered as one of the most widely distributed tick species ecto-parasitizing on livestock and causes fatal diseases with significant production loss. To address the problem of controlling acaricide resistant tick infestations on animals, attention has been paid to develop eco-friendly phyto-acaricides. The present study was undertaken to evaluate the comparative anti-tick activities of commercially available herbal and chemical products with chemically characterized phyto-formulations developed recently against field ticks and resistant tick lines of R. (B.) microplus. The chemical product Butox(®) Vet was found nearly passive against all the tested resistant tick lines. However, one of the commercial polyherbal product, Zerokeet(®) showed an efficacy (E%) of 41.8-75.4 % ([Formula: see text]) using recommended dilution (1:2) against field ticks and resistant tick lines. However, the other commercial product, Erina(®) EP has very limited efficacy against all the tested tick. In comparison, the newly developed phyto-formulation, NBA/13/B/2 and NAC-01 conferred an E% of 82.4-91.3 % ([Formula: see text]) and 62.3-94.6 % ([Formula: see text]), respectively, against tested resistant ticks. Results indicated higher marketing potentiality of newly developed formulation in the existing tick problem scenario.

  18. 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. (c) 2009. Published by Elsevier Ltd. All rights reserved.

  19. 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.

  20. Control of Rhipicephalus (Boophilus) microplus infestations by the combination of subolesin vaccination and tick autocidal control after subolesin gene knockdown in ticks fed on cattle.

    PubMed

    Merino, Octavio; Almazán, Consuelo; Canales, Mario; Villar, Margarita; Moreno-Cid, Juan A; Estrada-Peña, Agustín; Kocan, Katherine M; de la Fuente, José

    2011-03-09

    Tick subolesin was shown in immunization trials using the recombinant protein to protect hosts against tick infestations. In this study, we demonstrated that subolesin vaccination and release of ticks after subolesin knockdown by RNA interference (RNAi) could be used for the control of Rhipicephalus (Boophilus) microplus tick infestations in cattle and suggested that the combination of these methods could increase the efficacy of cattle tick control under some circumstances. The greatest tick control was obtained when both release of ticks after subolesin knockdown and vaccination were used concurrently. However, modeling results suggested that vaccine efficacy could be increased if at least 80% of the ticks infesting cattle correspond to subolesin-knockdown ticks. The results of this proof-of-concept trial demonstrated the efficacy of the sterile acarine technique (SAT) through production of subolesin-knockdown larvae by dsRNA injection into replete females for the control of R. microplus tick infestations, alone or in combination with subolesin vaccination. Copyright © 2011 Elsevier Ltd. All rights reserved.

  1. 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.

  2. [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.

  3. 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. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Acaricidal effect of essential oils from Lippia graveolens (Lamiales: Verbenaceae), Rosmarinus officinalis (Lamiales: Lamiaceae), and Allium sativum (Liliales: Liliaceae) against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

    PubMed

    Martinez-Velazquez, M; Rosario-Cruz, R; Castillo-Herrera, G; Flores-Fernandez, J M; Alvarez, A H; Lugo-Cervantes, E

    2011-07-01

    Acaricidal effects of three essential oils extracted from Mexican oregano leaves (Lippia graveolens Kunth), rosemary leaves (Rosmarinus officinalis L.), and garlic bulbs (Allium sativum L.) on 10-d-old Rhipicephalus (Boophilus) microplus (Canestrini) tick larvae were evaluated by using the larval packet test bioassay. Serial dilutions of the three essential oils were tested from a starting concentration of 20 to 1.25%. Results showed that both Mexican oregano and garlic essential oils had very similar activity, producing high mortality (90-100%) in all tested concentrations on 10-d-old R. microplus tick larvae. Rosemary essential oil produced >85% larval mortality at the higher concentrations (10 and 20%), but the effect decreased noticeably to 40% at an oil concentration of 5%, and mortality was absent at 2.5 and 1.25% of the essential oil concentration. Chemical composition of the essential oils was elucidated by gas chromatography-mass spectrometry analyses. Mexican oregano essential oil included thymol (24.59%), carvacrol (24.54%), p-cymene (13.6%), and y-terpinene (7.43%) as its main compounds, whereas rosemary essential oil was rich in a-pinene (31.07%), verbenone (15.26%), and 1,8-cineol (14.2%), and garlic essential oil was rich in diallyl trisulfide (33.57%), diallyl disulfide (30.93%), and methyl allyl trisulfide (11.28%). These results suggest that Mexican oregano and garlic essential oils merit further investigation as components of alternative approaches for R. microplus tick control.

  5. 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.

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

    USDA-ARS?s Scientific Manuscript database

    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....

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

    USDA-ARS?s Scientific Manuscript database

    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....

  8. Larvicidal activity of isolated compound 5-(2,4-dimethylbenzyl) pyrrolidin-2-one from marine Streptomyces VITSVK5 sp. against Rhipicephalus (Boophilus) microplus, Anopheles stephensi, and Culex tritaeniorhynchus.

    PubMed

    Saurav, Kumar; Rajakumar, Govindasamy; Kannabiran, Krishnan; Rahuman, Abdul Abdul; Velayutham, Kanayairam; Elango, Gandhi; Kamaraj, Chinnaperumal; Zahir, Abdul Abduz

    2013-01-01

    The aim of the present study was to assess the larvicidal property of marine actinobacterial compound 5-(2,4-dimethylbenzyl) pyrrolidin-2-one (DMBPO) extracted and isolated from Streptomyces VITSVK5 sp. tested against the larvae of Rhipicephalus (Boophilus) microplus Canestrini (Acari: Ixodidae), Anopheles stephensi Liston, and Culex tritaeniorhynchus Giles (Diptera: Culicidae). The isolate bacteria was taxonomically characterized, identified, and designated as Streptomyces VITSVK5 sp. The crude extract was loaded on silica gel column and eluted with chloroform:methanol. The isolated pure compound was analyzed by thin layer chromatography using chloroform and methanol as the solvent system and confirmed by high-performance liquid chromatography. The structure of the purified compound was established from infrared, ultraviolet, (1)H-nuclear magnetic resonance (NMR), (13)C-NMR, and mass spectral data. The chemical shift assignments obtained for the aliphatic compound from (1)H-NMR corresponding to the molecular formula C(13)H(17)NO. Bioassay-guided fractionation led to the isolation of compound which was identified as DMBPO. In the present study, Streptomyces VITSVK5 sp. crude extract and different fractions were tested against the larvae of parasites at the concentration of 1,000 ppm. Those fractions showing 100% mortality in 24 h alone was selected for further column chromatographic separation. The purified compound, C(13)H(17)NO, was tested in the concentrations of 500, 250, 125, 62.5, and 31.25 ppm and observed the percent larval mortality of 100, 70, 64, 40, and 28 against R. microplus; 100, 79, 63, 36, and 22 against A. stephensi; and 100, 84, 67, 42, and 27 against C. tritaeniorhynchus, respectively. The crude extract showed parasitic effects after 24 h of exposure at 1,000 ppm, and parasite mortality was observed against the larvae of R. microplus (LC(50) = 210.39 ppm, r (2) = 0.873); A. stephensi (LC(50) = 169.38 ppm, r (2) = 0

  9. Expression profiling, gene silencing and transcriptional networking of metzincin metalloproteases in the cattle tick, Rhipicephalus (Boophilus) microplus.

    PubMed

    Barnard, Annette-Christi; Nijhof, Ard M; Gaspar, Anabella R M; Neitz, Albert W H; Jongejan, Frans; Maritz-Olivier, Christine

    2012-05-25

    Tick proteins functioning in vital physiological processes such as blood meal uptake, digestion and reproduction are potential targets for anti-tick vaccines, since vaccination could inhibit these essential functions and ultimately affect tick survival. In this study we identified metzincin metalloproteases from Rhipicephalus microplus as potential vaccine candidates since they are implicated as essential to blood-cavity formation, bloodmeal digestion and reproduction in ixodid ticks. Eight transcripts encoding proteins that contain the characteristic metzincin zinc-binding motif HEXXHXXG/NXXH/D and a unique methionine containing "methionine-turn" were identified from native and in-house assembled R. microplus expressed sequence tag (EST) databases. These were representative of five reprolysin-like and three astacin-like metzincin metalloproteases. Reverse transcription-PCR analysis indicated that the reprolysins were most abundantly expressed in the salivary glands, whereas the astacins were most abundant in the midgut and ovaries. In vivo gene silencing was performed to assess a possible phenotype of these metalloproteases during adult female R. microplus blood feeding and reproduction. RNA interference (RNAi) against two of the reprolysins and one of the astacins significantly affected the average egg weight and oviposition rate. Evidently, this reverse genetic approach enabled the evaluation of the overall vital impact of tick proteins. Finally, integrated real time-PCR studies also revealed an extensive cross organ network between the R. microplus metzincin transcripts, supporting the use of a combinatorial metzincin-based anti- R. microplus vaccine. Copyright © 2011 Elsevier B.V. All rights reserved.

  10. 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.

  11. 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.

    PubMed

    Lovis, Leonore; Guerrero, Felix D; Miller, Robert J; Bodine, Deanna M; Betschart, Bruno; Sager, Heinz

    2012-12-01

    Resistance to synthetic pyrethroids (SP) in the cattle tick Rhipicephalus (Boophilus) microplus is widespread throughout its distribution area. Three single nucleotide substitutions identified in Domains II and III of the sodium channel gene of R. (B.) microplus are known to be associated with target site pyrethroid resistance. We developed a multiplex PCR using allele-specific primers to amplify wild type or mutated genotypes of the three mutations simultaneously. This assay was used to screen tick samples originating from Brazil, Argentina, Mexico, South Africa and Australia whose phenotype to flumethrin and cypermethrin had been determined by the use of the Larval Tarsal test (LTT) or the Larval Packet Test (LPT). These mutations were found to have distinct geographical distributions and result in different resistance phenotypes. The L64I Domain II mutation conferring resistance to several SP compounds was found in all the Brazilian, Argentinean and Australian populations and in one South African population, with frequencies between 38% and 100% in flumethrin and cypermethrin resistant populations. In contrast, this mutation was not found in samples from Mexico, while the Domain III mutation was found exclusively in this country. The G72V Domain II flumethrin-specific mutation was found in a single Australian population, with a very low resistant allele frequency (3%). The homozygous resistant RR genotype of the L64I Domain II mutation correlated significantly with the survival rates at the discriminating doses of flumethrin and cypermethrin. This survey shows the widespread distribution of the L64I Domain II mutation and provides evidence of its geographic separation from the Domain III mutation.

  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

    PubMed Central

    Lovis, Leonore; Guerrero, Felix D.; Miller, Robert J.; Bodine, Deanna M.; Betschart, Bruno; Sager, Heinz

    2012-01-01

    Resistance to synthetic pyrethroids (SP) in the cattle tick Rhipicephalus (Boophilus) microplus is widespread throughout its distribution area. Three single nucleotide substitutions identified in Domains II and III of the sodium channel gene of R. (B.) microplus are known to be associated with target site pyrethroid resistance. We developed a multiplex PCR using allele-specific primers to amplify wild type or mutated genotypes of the three mutations simultaneously. This assay was used to screen tick samples originating from Brazil, Argentina, Mexico, South Africa and Australia whose phenotype to flumethrin and cypermethrin had been determined by the use of the Larval Tarsal test (LTT) or the Larval Packet Test (LPT). These mutations were found to have distinct geographical distributions and result in different resistance phenotypes. The L64I Domain II mutation conferring resistance to several SP compounds was found in all the Brazilian, Argentinean and Australian populations and in one South African population, with frequencies between 38% and 100% in flumethrin and cypermethrin resistant populations. In contrast, this mutation was not found in samples from Mexico, while the Domain III mutation was found exclusively in this country. The G72V Domain II flumethrin-specific mutation was found in a single Australian population, with a very low resistant allele frequency (3%). The homozygous resistant RR genotype of the L64I Domain II mutation correlated significantly with the survival rates at the discriminating doses of flumethrin and cypermethrin. This survey shows the widespread distribution of the L64I Domain II mutation and provides evidence of its geographic separation from the Domain III mutation. PMID:24533283

  13. 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.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

  17. [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.

  18. The productivity effects of cattle tick (Boophilus microplus) infestation on cattle, with particular reference to Bos indicus cattle and their crosses.

    PubMed

    Jonsson, N N

    2006-04-15

    In response to uncertainty among cattle producers in Australia regarding the need to treat Bos indicus and B. indicus crossbreeds, the scientific literature relating to the productivity effects of Boophilus microplus on cattle of all breeds was reviewed. Estimates of the mean effect of each engorging tick (damage coefficient, d) were made from a simple analysis of the reported data. On average, each engorging female tick is responsible for the loss of 1.37 +/- 0.25 g bodyweight in B. taurus cattle. The comparable value for B. taurusxB. indicus cattle is 1.18 +/- 0.21 g/engorging tick. These values were not statistically significantly different, indicating that if a threshold approach to tick control were taken, then the threshold number of standard ticks would be the same regardless of cattle genotype. No studies provided useable estimates of the effect of tick infestation on pure B. indicus cattle. An economic threshold for treatment, below which acaricide application is not beneficial, can be predicted, using known values for the cost of acaricide application and the price of beef. However, the application of a threshold approach to control has not been embraced by government advisers and runs contrary to the accepted principals of strategic control programs.

  19. 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.

  20. 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.

  1. 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

  2. Identification and structural-functional analysis of cyclin-dependent kinases of the cattle tick Rhipicephalus (Boophilus) microplus.

    PubMed

    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.

  3. 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

  4. [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%).

  5. Differences in the life cycles between a vaccine strain and an unmodified strain of Babesia bovis (Babes, 1889) in the tick Boophilus microplus (Canestrini).

    PubMed

    Stewart, N P

    1978-11-01

    Developmental forms of 2 strains of Babesia bovis (Babes) were studied in the tick vector Boophilus microplus (Canestrini). One strain (designated T) was shown to be infective for the tick, and the other (NT) to have lost infectivity for the tick, because of repeated blood passaging in cattle. Parasites of the 2 strains in gut contents of adult female ticks were similar during the first 16 h post-repletion (PR), but thereafter their structure differed. From 16-64 h PR, the majority of T strain parasites were spherical and without processes. During the next 32 h elongate forms and vermicules developed. Fission bodies were seen within epithelial cells of the gut by 96 h PR. T-strain parasites in gut contents decreased in number from approximately 96 h and were difficult to find at 144 h, the time of the final observation. In contrast, NT strains parasites were plentiful throughout the period of observation. They were predominantly spherical, ranging in diameter from 1.5 to 15 micron. Forms with obvious processes measuring up to 81 micron in length were seen in large numbers at seemingly regular intervals from 16-44 h PR, suggesting that a process of development and divisions was being repeated. No vermicules or fission bodies were seen. T-strain, but not NT strain parasites, were seen in hemolymph and ova of the ticks and in their larval progeny. It is suggested that continuous blood passaging of the NT strain had resulted in selection of parasites incapable of penetrating gut epithelial cells of the tick.

  6. Efficacy of amitraz applied as a dip against an amitraz-resistant strain of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) infested on cattle.

    PubMed

    Davey, Ronald B; Miller, Robert J; George, John E

    2008-03-25

    Selection pressure with the acaricide, amitraz was applied to a strain of Rhipicephalus (Boophilus) microplus (Canestrini) in 13 out of 18 generations, resulting in a 28.7-fold increase in resistance, indicating a shift in the phenotypic composition of the ticks, from a heterogenous mixture of both susceptible and resistant ticks when selection began to a homogenous composition of predominantly resistant ticks when selection ended. Resistant larvae of the last selected generation were infested on cattle three separate times at 20, 13, and 6 d (7 d intervals) before being dipped in a total immersion vat charged at 0.02, 0.044, and 0.096% active ingredient (AI) amitraz. There was a dose related, though not always significant, response to amitraz whereby each increase in concentration produced increased adverse effects. The number of ticks and index of fecundity (IF) of females recovered from cattle treated at 0.02 and 0.044% AI were not different from that of untreated ticks. However, female weight and egg mass weight of females recovered from cattle treated at 0.044% were lower than untreated females or females treated at 0.02% AI. All measured parameters obtained from ticks recovered from cattle treated at 0.096% AI were significantly lower than the untreated control group, but tick numbers and IF were not different from the other treated groups. At 0.02% AI the level of control was significantly lower (34.6%) than treatment at 0.096% AI (81.2%), while the level of control at 0.044% AI (57.6%) was intermediate between the other treated groups. Therefore, none of the amitraz treatments provided the required 99% control necessary for use in the U.S. Cattle Fever Tick Eradication Program, and the estimated line derived from regression analysis showed that a concentration of ca. 0.2% AI amitraz would be needed to achieve 99% control of the resistant ticks.

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

    DTIC Science & Technology

    2013-03-28

    melanogaster; DMSO, dimethyl sulfoxide; DTNB, 5,5’-dithiobis-(2-nitro) benzoic acid ; hAChE, recombi- nant human AChE; IC50, inhibitory concentration needed...bendiocarb) are highly potent inhibitors of both rBmAChE1 and rPpAChE (Table 2). For both rBmAChE and rPpAChE, Table 1 Amino acid substitutionsa in recombinant...BmAChE1 constructs of R. (B.) microplus. BmAChE1 Amino acid Substitution (wt) Deutch #5 (OP-S) Muñoz #4 (OP-S) San Roman #4 (OP-R) San Roman #11 (OP

  8. 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. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Contributions of the hair sheep breed Santa Ines as a maintenance host for Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in Brazil.

    PubMed

    Garcia, Marcos Valério; Andreotti, Renato; Reis, Fernando Alvarenga; Aguirre, André de Abreu Rangel; Barros, Jacqueline Cavalcante; Matias, Jaqueline; Koller, Wilson Werner

    2014-11-18

    Hair sheep breeds are a new, cost-effective option for the diversification of livestock in the Midwest region of Brazil. They are grazed extensively with cattle as well as in isolation in small areas. Hair sheep breeds are vulnerable to infestation by parasites such as the cattle tick, Rhipicephalus microplus, which causes various types of damage and can transmit diseases. In this study, Santa Inês hair sheep were naturally infested in an area contaminated by infested cattle and then monitored to determine the ability of these animals to maintain the local tick population in the absence of cattle. After engorged tick females of each generation fell off, the animals were placed in another pasture and were returned only after larvae reappeared in the original pasture. Tick counts were performed every ten days for three generations of sheep, and average infestations per animal of 34, 12 and 4 ticks were observed for each successive generation. These numbers suggest the acquisition of resistance; however, additional studies are needed to ensure resistance is achieved. The average length of the parasitic phase for each generation of ticks was 25 days. We concluded that this hair sheep breed, even if kept separate from cattle, is able to maintain tick populations for at least three generations, although a gradual decrease in the population levels of R. microplus over three generations was observed. We also detected two positive cases of Anaplasma spp. Therefore, it appears that the Santa Inês hair sheep breed contributes to the circulation of this bacterium among other ruminants.

  10. 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.

  11. In-vitro efficacy of a botanical acaricide and its active ingredients against larvae of susceptible and acaricide-resistant strains of Rhipicephalus (Boophilus) microplus Canestrini (Acari: Ixodidae).

    PubMed

    Singh, Nirbhay K; Miller, Robert J; Klafke, Guilherme M; Goolsby, John A; Thomas, Donald B; Leon, Adalberto A Perez de

    2017-09-14

    Ticks and tick-borne diseases are a major constraint for a sustainable cattle industry in the tropical and subtropical regions of the world. The development of resistance to most of the commonly used acaricides has led to an assessment of herbal products for acaricidal activity as an eco-friendly tick control alternative. A botanical product, Essentria(®) IC-3 insect concentrate containing rosemary oil (10%), geraniol (5%) and peppermint oil (2%), acts on target pests by blocking octopamine. Essentria(®) IC-3 and its active components were evaluated for larvicidal activity against several susceptible and acaricide-resistant strains of the cattle fever tick, Rhipicephalus (Boophilus) microplus Canestrini by Larval Packet test using 14-21 d old unfed larvae. The efficacy was assessed by measuring percent larval mortality and estimating lethal concentrations at 50% (LC50) and 95% (LC95) with 95% confidence limits (CL) using probit analysis. The LC50 and LC95 (95% CL) values for Essentria(®) IC-3 against the susceptible strain were estimated as 0.647% (0.59-0.69) and 1.033% (0.94-1.19), respectively, whereas, LC50 and LC95 values for other strains were variable, ranging from 0.597-1.674% and 0.927-2.236%, respectively. Among the various active ingredients, the larvicidal property of Essentria(®) IC-3 seem to be attributable mainly to geraniol and the LC50 and LC95 (95% CL) values for geraniol against the susceptible Deutch strain were estimated as 0.656% (0.61-0.69) and 1.114% (1.03-1.25), respectively. The comparison of LC50 and LC95 values of acaricide-resistant strains showed susceptibility comparable to Deutch against geraniol except for the Las Palmas strain. We report a low level of resistance in some of the acaricide-resistant strains against the herbal acaricide in the cattle tick for the first time, possibly due to cross-resistance to chemical acaricides. Copyright © 2017. Published by Elsevier GmbH.

  12. Immune Response of Bos indicus Cattle against the Anti-Tick Antigen Bm91 Derived from Local Rhipicephalus (Boophilus) microplus Ticks and Its Effect on Tick Reproduction under Natural Infestation.

    PubMed

    Lambertz, Christian; Chongkasikit, Natthaphon; Jittapalapong, Sathaporn; Gauly, Matthias

    2012-01-01

    Antigens of anti-tick vaccines are more efficacious for homologous challenge with local tick strains. cDNA clones encoding for Bm91 from local Rhipicephalus (Boophilus) microplus strains were developed to immunize Bos indicus cattle under field conditions. Three groups of six animals each were injected with the antigen Bm91, saline, and adjuvant, respectively. Animals were immunized three times at 3-week intervals and a fourth time after six months. The anti-Bm91 antibody level, measured by ELISA, was monitored for 7 months and the reproductive performance of naturally infested R. (B.) microplus was determined. Bm91-immunized animals developed a strong immune response expressed by high anti-Bm91 levels remaining on high levels until the end of the study. Western blot analysis confirmed that Bm91 is immunogenic. Compared to control animals, the reproductive efficiency index and the egg viability were 6% and 8%, respectively, lower in the Bm91 group (P < 0.05). In conclusion, it was demonstrated that Bm91 induced a long-lasting immune response. However, the effect on the tick reproduction was not sufficient for an efficient tick control. Further studies under field conditions are warranted to enhance the effect on the tick reproduction by optimizing the immunization regimen, alone or in combination with other vaccine candidate antigens.

  13. 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.

  14. 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. © 2016 The Royal Entomological Society.

  15. 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.

  16. 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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  1. Suppression of Rhipicephalus (Boophilus) microplus under natural South Texas conditions using Y-Tex® XP820™ ear tags on pastured cattle

    USDA-ARS?s Scientific Manuscript database

    Pastured cattle with ear tags containing 8% abamectin synergized with 20% piperonil butoxide were evaluated for efficacy against Rhipicephalus microplus ticks during a 40-week study conducted under natural South Texas conditions. In the first 16 weeks, which occurred during the spring and early summ...

  2. Strategic control of Rhipicephalus (Boophilus) microplus infestation on beef cattle grazed in Panicum maximum grasses in a subtropical semi-arid region of Argentina.

    PubMed

    Morel, Nicolás; Signorini, Marcelo L; Mangold, Atilio J; Guglielmone, Alberto A; Nava, Santiago

    2017-09-01

    The aim of this work was to test the efficacy of strategic control methods of Rhipicephalus microplus infestation on beef cattle grazed in Panicum maximum grasses in northwestern Argentina. Also, an analysis to discern how the R. microplus population was distributed amongst cattle was also performed to determine if partial selective treatment or cull the small proportion of more heavily infested animals are feasible options to control this tick. The strategic scheme of treatments was designed to act on the small 1st generation of R. microplus in early spring and prevent in that way the appearance of the annual peak of abundance of R. microplus in summer and autumn. Animals of the group 1 were treated with ivermectin 3.15% on day 0 (25th September 2015), with fluazuron on day 32 (27th October 2015) and with fipronil on day 75 (9th December 2015). Animals of group 2 formed the control group. The overall effect of the treatments was positively significant. The number of ticks observed on the control group was significantly higher than the number of ticks observed on the treated group in all post-treatment counts (P<0.01), with the only exception of the count of March. The distribution of parasites among cattle in all counts was adjusted to the negative binomial distribution, but a temporal variation in the tick aggregation levels associated to changes in tick abundance was found. The higher the abundance of R. microplus, the lower the aggregation. It was found that the steers (15.8% of the total number of animals evaluated) belonging to the high infestation group accounted for 23.0% of the total ticks. The strategic control method evaluated during this study provides a remarkable overall effect against R. microplus because it significantly reduces the tick infestation on cattle with only three applications of acaricides in one-year period. The analyses of tick distribution amongst cattle suggest that partial selective treatment and culling do not represent feasible

  3. 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

  4. 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

  5. Potential synergistic effect of Melia azedarach fruit extract and Beauveria bassiana in the control of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in cattle infestations.

    PubMed

    Sousa, Lorena Alessandra Dias; Pires, Hélio Bernardes; Soares, Sara Fernandes; Ferri, Pedro Henrique; Ribas, Patricia; Lima, Eliane Martins; Furlong, John; Bittencourt, Vânia Rita Elias Pinheiro; Perinotto, Wendell Marcelo de Souza; Borges, Lígia Miranda Ferreira

    2011-02-10

    The use of a concentrate emulsion of Melia azedarach green fruits and a suspension of the fungus Beauveria bassiana was evaluated in the control of Rhipicephalus microplus on artificially infested cattle. The evaluation was conducted following the protocol established by the Brazilian Agriculture Ministry. Five groups of 4 or 5 animals were allocated to one of the following treatments: emulsion concentrate of M. azedarach at 0.25% (T AZED 0.25%), emulsion concentrate of M. azedarach at 0.5% (T AZED 0.5%), B. bassiana at 2.4 × 10(8) conidia (T BASS), association of the concentrate of M. azedarach at 0.25% with B. bassiana at 2.4 × 10(8) conidia (T AZED 0.25%+BASS), and control (untreated). The association of the two compounds provided better results than any one isolated treatment, indicating compatibility or perhaps a synergy between M. azedarach and B. bassiana. This treatment resulted in fewer engorged females (129 ± 70) than in the control group (233 ± 82), showing high performance against all developmental stages of the tick. Results revealed an apparent synergistic effect of M. azedarach and B. bassiana in the control of R. microplus that should be further investigated. Copyright © 2010 Elsevier B.V. All rights reserved.

  6. Acaricidal effects of fluazuron (2.5 mg/kg) and a combination of fluazuron (1.6 mg/kg) + ivermectin (0.63 mg/kg), administered at different routes, against Rhipicephalus (Boophilus) microplus parasitizing cattle.

    PubMed

    Gomes, Lucas Vinicius Costa; Lopes, Welber Daniel Zanetti; Cruz, Breno Cayeiro; Teixeira, Weslen Fabricio; Felippelli, Gustavo; Maciel, Willian G; Bichuette, Murilo Abud; Ruivo, Maycon A; Alcantara Colli, Marcos Henrique; Carvalho, Rafael Silveira; Martinez, Antonio Campanha; Soares, Vando Edésio; da Costa, Alvimar José

    2015-06-01

    The present study aimed to evaluate the acaricidal efficacy of fluazuron (2.5 mg/kg), administered as a pour-on, in comparison to an injectable formulation containing fluazuron (1.6 mg/kg) + ivermectin (0.63 mg/kg), against Rhipicephalus (Boophilus) microplus in naturally and experimentally infested cattle. Two studies were conducted with different tick strains, one with artificial infestations (Stall Test, using leight animals per group) and one with natural infestations (utilizing ten animals per group). In both studies, the animals were randomized, according to average tick counts performed on days -3, -2 and -1, into four groups: T01, negative control (saline solution); T02, pour-on fluazuron (2.5 mg/kg); T03: subcutaneous fluazuron (1.6 mg/kg) + ivermectin (0.63 mg/kg); and T04 subcutaneous ivermectin (0.63 mg/kg). Based on obtained results, and considering the utilized tick strains, it was possible to conclude that the pour-on fluazuron (2.5 mg/kg) formulation demonstrated high acaricidal efficacy, with protection periods ranging from 49 to 77 days against Rhipicephalus (Boophilus) microplus. On the other hand, for the injectable fluazuron (1.6 mg/kg) + ivermectin (0.63 mg/kg) formulation, it was not possible to observe elevated anti-R. (B.) microplus effect on both artificial and experimental infestation studies. Results observed for this combination were similar or inferior to those obtained by subcutaneous ivermectin (0.63 mg/kg). Future studies with this formulation containing fluazuron (1.6 mg/kg) + ivermectin (0.63 mg/kg), regarding pharmacokinetic and/or bioavailability profiles, or even studies analyzing both this active principles separately, are needed, seeking to better understand the effects of such combination against Rhipicephalus (Boophilus) microplus parasitizing cattle. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. 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.

  8. Rhipicephalus microplus strain Deutsch, 10 BAC clone sequences

    USDA-ARS?s Scientific Manuscript database

    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...

  9. Susceptibility of Rhipicephalus (Boophilus) microplus to fluazuron (2.5mg/kg) and a combination of novaluron (2.0mg/kg)+eprinomectin (0.36mg/kg) in field studies in Brazil.

    PubMed

    Maciel, Willian Giquelin; Lopes, Welber Daniel Zanetti; Gomes, Lucas Vinicius Costa; Cruz, Breno Cayeiro; Felippelli, Gustavo; Santos, Isabella Barbosa Dos; Borges, Fernando de Almeida; Gonçalves, Walter Antonio; Scarpa, Alexandre Braga; Nicaretta, João Eduardo; Bastos, Thiago Souza Azeredo; da Costa, Alvimar José

    2016-12-01

    The present study aimed to determine the susceptibility of 32 R. (B.) microplus populations from Southeast, Midwest and South regions of Brazil, to fluazuron (2.5mg/kg), administered topically (pour-on). Additionally, five populations (Southeast and Midwest regions) of the southern cattle tick were evaluated using in vivo field studies, regarding their susceptibility to a new combination of novaluron (2.0mg/kg)+eprinomectin (0.36mg/kg), administered subcutaneously, compared with two positive controls (fluazuron 2.5mg/kg and eprinomectin 0.5mg/kg), both administered topically (pour-on). Selected bovines were allocated to treatment groups on day 0, and block formation was based on arithmetic means of female ticks (4.5-8.0mm long) counted on three consecutive days (-3, -2 and -1). To evaluate therapeutic and residual efficacies of these formulations, tick counts (females ranging from 4.5 to 8.0mm long) were performed on days 3, 7 and 14 post-treatment, continuing on a weekly basis until the end of each experiment. Results obtained throughout this study, utilizing field efficacy trials, allowed us to conclude that four R. (B.) microplus populations (including two in the Southeast and two in the Midwest regions) could be diagnosed as resistant, or with low susceptibility, to fluazuron (2.5mg/kg). Such fact was detected in farms where owners applied products containing this active component on cattle for at least five years, with treatment intervals of 30-55days during the rainy season. Nonetheless, in vitro studies should be performed in order to reinforce in vivo results obtained on the present study. Regarding efficacy indexes obtained by the association of eprinomectin and the novel molecule novaluron against R. (B.) microplus, none of the trials managed to obtain efficacies superior to 48%. Such results, allied to data obtained by different researchers and previously published in literature, reinforce the perception that maybe these formulations containing novaluron

  10. Immunoregulation of bovine macrophages by factors in the salivary glands of Rhipicephalus microplus

    USDA-ARS?s Scientific Manuscript database

    The southern cattle fever tick, Rhipicephalus (Boophilus) microplus, is a vector of the apicomplexan protozoa Babesia bovis and B. bigemina, which cause the often fatal disease bovine babesiosis, also known as cattle fever. Although the Cattle Fever Tick Eradication Program (CFTEP) succeeded in elim...

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  15. 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 infested or exposed animals prohibited. 72.1 Section 72.1 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE INTERSTATE TRANSPORTATION OF ANIMALS (INCLUDING POULTRY) AND ANIMAL PRODUCTS TEXAS (SPLENETIC) FEVER...

  16. 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...

  17. 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...

  18. 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...

  19. Characterization of a novel cystatin type 2 from Rhipicephalus microplus midgut.

    PubMed

    Cardoso, Thyago H S; Lu, Stephen; Gonzalez, Boris R G; Torquato, Ricardo J S; Tanaka, Aparecida S

    2017-09-01

    The Rhipicephalus (Boophilus) microplus is an exclusive bovine ectoparasite responsible for the transmission of pathogens that decrease meat, leather and milk productions. Cattle vaccination is an alternative to control tick infestations, but the discovery of potential antigens is still a challenge for researchers. Recently, our group performed a midgut transcriptome of engorged R. microplus tick, and out of 800 ESTs sequences one cystatin-coding sequence was identified and named Rmcystatin-4. In order to understand the physiological role of Rmcystatin-4, the aim of this work was the expression, purification and functional characterization of a novel type 2 cystatin from the tick R. microplus. Rmcystatin-4 gene expression was identified mostly in tick midgut suggesting its possible role in blood digestion control. Our data showed that rRmcystatin-4 was successfully expressed in active form using Pichia pastoris system and the purified inhibitor presented high selectivity to BmCl-1 (Ki = 0.046 nM). Moreover, rRmcystatin-4 was able to impaired BmCl-1 activity towards bovine hemoglobin. Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  20. 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

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

    PubMed

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

    2016-07-01

    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 amino acid 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. Published by Elsevier GmbH.

  2. A simulation study of the effects of acaricides and vaccination on Boophilus cattle-tick populations.

    PubMed

    Lodos, J; Ochagavia, M E; Rodriguez, M; De La Fuente, J

    1999-01-01

    This paper describes a simulation model to evaluate different control strategies for Boophilus microplus. The model combines a dynamic life-history module for tick-population dynamics with other modules for vaccination, sterile-hybrid larval release and use of acaricide dipping vats. The tick life-history module considers the cattle's nutritional level and allows for distribution of ticks by age at all stages of growth. Appropriately, the model was sensitive to host resistance and to host-nutritional status. The validity of the life-history module--as well as that of the vaccination and acaricide dipping--vats modules--was demonstrated by comparing simulated and real data for several geographical locations in Cuba and Brazil. Optimum tick-control strategies for the first year of vaccination were designed and the effect of long-term vaccination on tick population was also studied.

  3. Histoarchitecture of the Ovary of Rhipicephalus (Boophilus) annulatus during Pre- and Postengorgement Period

    PubMed Central

    Kanapadinchareveetil, Sreelekha; Chandrasekhar, Leena; Gopi, Jyothimol; Ranjan Lenka, Dibya; Vasu, Aswathi; KGopalan, Ajith Kumar; Nair, Suresh N.; Juliet, Sanis; Ghosh, Srikanta

    2015-01-01

    The present communication describes the detailed day wise study of histological changes of the ovary of Rhipicephalus (Boophilus) annulatus in the postengorgement period together with the systematic classification of their oocytes. The ovary of R. (B.) annulatus is panoistic type with an asynchronous development of oocytes. All the stages (II, III, IV, and V) of oocytes except stage I were similar to R. (B.) microplus. The stage I oocytes showed basophilia, which was not reported earlier in other species of ticks. Day wise changes were in the form of presence of oogonia in partially fed and day one engorged adults, considerable degeneration of oocytes on day two, emergence of new wave of oocytes on day three, presence of mature oocytes up to day eight, and complete degeneration of ovarian tissue from day eight onwards. The degenerative changes in the ovary appeared initially in the oocytes followed by germinal epithelium. PMID:25664337

  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. Rhipicephalus microplus salivary gland molecules induce differential CD86 expression in murine macrophages

    PubMed Central

    2010-01-01

    Background Tick parasitism is a major impediment for cattle production in many parts of the world. The southern cattle tick, Rhipicephalus (Boophilus) microplus, is an obligate hematophagous parasite of domestic and wild animals that serves as vector of infectious agents lethal to cattle. Tick saliva contains molecules evolved to modulate host innate and adaptive immune responses which facilitates blood feeding and pathogen transmission. Tick feeding promotes CD4 T cell polarization to a Th2 profile usually accompanied by down-regulation of Th1 cytokines through as yet undefined mechanisms. Co-stimulatory molecules on antigen presenting cells are central to development of T cell responses including Th1 and Th2 responses. Tick induced changes to antigen presenting cell signal transduction pathways are largely unknown. Here we document the ability of R. microplus salivary gland extracts (SGE) to effect differential CD86 expression. Results We examined changes in co-stimulatory molecule expression in murine RAW 264.7 cells in response to R. microplus SGE exposure in the presence of the toll-like receptor 4 (TLR4) ligand, LPS. After 24 hrs, CD86, but not CD80, was preferentially up-regulated on mouse macrophage RAW 264.7 cells when treated with SGE and then LPS, but not SGE alone. CD80 and CD40 expression was increased with LPS, but the addition of SGE did not alter expression. Higher concentrations of SGE were less effective at increasing CD86 RNA expression. The addition of mitogen or extracellular kinase (MEK) inhibitor, PD98059, significantly reduced the ability for SGE to induce CD86 expression, indicating activation of MEK is necessary for SGE induced up-regulation. Conclusions Molecules in SGE of R. microplus have a concentration-dependent effect on differential up-regulation of CD86 in a macrophage cell line activated by the TLR4 ligand, LPS. This CD86 up-regulation is at least partially dependent on the ERK1/2 pathway and may serve to promote Th2 polarization

  6. Survey of cattle tick, Riphicephalus (Boophilus) microplus, resistance to amitraz and deltamethrin in New Caledonia.

    PubMed

    Petermann, J; Cauquil, L; Hurlin, J C; Gaia, H; Hüe, T

    2016-02-15

    The evolution of tick resistance to acaricides in New Caledonia was followed in two ways: through two large scale surveys in 1998 and 2014 and through the routine analysis of samples of ticks with suspected resistance. The results of the two approaches were qualitatively similar though analysis of ticks with suspected resistance gave higher frequencies of resistance, as expected of a biased sample. Resistance tests using a larval packet test have been conducted since 1993 for deltamethrin and 2003 for amitraz. Deltamethrin was used in country-wide control from 1986 to 2003 and amitraz since 1996. This study analyzed the variation of resistance parameters such as lethal concentration 50 (LC50) and the resistance factor over 21 years for deltamethrin and LC99 over 11 years for amitraz. There was an increase in deltamethrin's annual mean LC50 from 1993 to 2004 when it reached 2.9 g/l and then a progressive decrease, reaching a level consistently below 1g/l since 2007. Even though breeders stopped using deltamethrin in New Caledonia in 2003, the percentage of susceptible strains has remained below 30% since 1998. Amitraz's LC99 mean increased significantly from 0.31 g/l in 2003 to 2.96 g/l in 2014. Whereas all tested strains in 2003 were susceptible, only 40% of strains tested at the request of farmers were susceptible in 2014. The recent territory-wide survey showed that 76.7% of strains are still susceptible. This study established that resistance to amitraz has developed slowly in New Caledonia. A reversion phenomenon may have occurred concerning deltamethrin resistance, visible through the decrease of LC50 mean and the decreased proportion of very resistant strains; however the proportion of susceptible strains remains at a low level and there is anecdotal evidence that high resistance can re-emerge rapidly. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. 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

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

    USDA-ARS?s Scientific Manuscript database

    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...

  9. 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.

  10. A new family of cytochrome P450 genes (CYP41) from the cattle tick, Boophilus microplus.

    PubMed

    Crampton, A L; Baxter, G D; Barker, S C

    1999-09-01

    We isolated and sequenced a cytochrome P450 (CYP) gene that is sufficiently different from other CYP genes that a new CYP family, CYP41 was created. CYP41 encodes a protein of 518 residues and is most similar to genes from the family CYP3; it is 36% identical to CYP3A2 and 34% identical to CYP3A28. We hypothesise that CYP41 encodes an enzyme that metabolizes xenobiotic compounds i.e. compounds that are foreign to the cattle tick. The phylogenetic position of CYP41 could not be resolved because of the high level of sequence divergence at both the nucleotide and amino acid levels.

  11. Observation on the composition and biosynthesis of egg wax lipids in the cattle tick, Boophilus microplus.

    PubMed

    Booth, T F

    1992-05-01

    The biosynthesis of wax lipids by Gené's organ, the egg waxing organ in ticks, was investigated. Gené's organ, a complex dermal gland system, applies a superficial wax layer to the eggs during oviposition which prevents desiccation and is essential for egg viability. The detailed anatomy and histology of the three gland cell types are unambiguously described. Serial sectioning of ticks showed that all three gland cell types are capable of contributing to the egg wax. The wax synthetic ability of these three gland types was characterized. The composition of wax lipids extracted from the surface egg wax, and from the three types of wax gland dissected from ovipositing ticks, was analysed using thin-layer and gas-liquid chromatography. Injection of ovipositing ticks with radiolabelled acetate resulted in the incorporation of the label into wax lipids by gland cells of Gené's organ. The egg wax was a complex mixture of long-chain alkanes and fatty acid esters. The gland cells contained a greater proportion of shorter chain alkanes than were present in the egg surface wax. Some unsaturated long-chain fatty acids were present, and these were more abundant in the gland cells than in the surface wax of oviposited eggs, suggesting that oxidation occurs after oviposition. The results confirm that the tubular glands, acinar accessory glands and lobular glands of Gené's organ all contribute to the egg waxes, although the lipid components differed in relative abundance. The results are also consistent with alkane synthesis from fatty acids in Gené's organ by a chain-elongation-decarboxylation pathway.

  12. 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. This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.

  13. 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

  14. Rhipicephalus microplus and Dermacentor nitens (Acari: Ixodidae) Coparasitize White-Tailed Deer on St. John, U.S. Virgin Islands.

    PubMed

    Nelson, Suzanne L; Durden, Lance A; Reuter, Jon D

    2017-09-01

    Ticks parasitizing introduced white-tailed deer, Odocoileus virginianus (Zimmermann), on St. John, U.S. Virgin Islands, were recorded during and after drought conditions. Tick infestation prevalences were 22% at the start of the drought (July 2015), 66% at the height of the drought (March 2016), and 35% after the drought had ended (July 2016; n = 67 deer). Samples of ticks from 22 tranquilized deer in July 2016 revealed the presence of two species, the southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini), and the tropical horse tick, Dermacentor (Anocentor) nitens Neumann. Both tick species have considerable veterinary importance, especially for cattle and horses, respectively, as nuisance biters and also as vectors of parasitic piroplasms or of Anaplasma marginale Theiler. All 22 deer examined were infested by R. microplus, whereas 14 (64%) of the samples also included specimens of D. nitens. Because of the large numbers of ticks recorded, wild deer on St. John could develop associated health problems (pruritis, alopecia, anemia, low weight gain, tick-borne pathogens and parasites) and could also serve as a source of these ticks for cattle and horses. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  15. Influence of acaricide resistance on cattle-fever tick (Boophilus spp.) infestations in semi-arid thornshrublands: a simulation approach.

    PubMed

    Corson, M S; Teel, P D; Grant, W E

    2001-01-01

    Cattle-fever tick (Boophilus microplus and B. annulatus) populations that develop acaricide resistance become more difficult to control or eradicate. We used a simulation model to assess the direct and indirect effects of interactions among season, habitat type, grazing strategy, and acaricide resistance on the ability to eradicate Boophilus infestations in semi-arid thornshrublands of Texas, USA. Season of infestation appeared to have the strongest effect, with infestations begun on 27 September (autumn) tending to die out sooner than those begun on 1 March (spring) and to remain undetected. Habitat type had the next strongest effect, with infestations surviving much longer as canopy cover increased from uncanopied buffelgrass (Cenchrus ciliaris) habitats to mesquite (Prosopis glandulosa)-canopied grass habitats. Acaricide resistance had a moderate effect; as expected, highly resistant tick populations survived longer than those with no acaricide resistance. The importance of grazing strategy varied with changes in habitat type: as canopy cover increased, infestation duration increased faster under continuous grazing than under rotational grazing strategies. Importance of grazing strategy also varied with acaricide resistance: detected tick populations with no and slight acaricide resistance subjected to acaricide treatments tended to survive longer under rotational grazing than continuous grazing, due to reduced contact with a treated host. Populations with moderate and high resistance behaved more like untreated populations, tending to survive longer under continuous, rather than rotational, grazing, because they experienced less mortality on a treated host. Assuming acaricide treatments at 2-week intervals and maintenance of cattle in infested pastures, results indicate that, for each habitat type, infesting ticks have a threshold of acaricide resistance below which one can eradicate them faster with continuous grazing than with rotational grazing. As canopy

  16. Widespread movement of invasive cattle fever ticks (Rhipicephalus microplus) in southern Texas leads to shared local infestations on cattle and deer

    PubMed Central

    2014-01-01

    Background Rhipicephalus (Boophilus) microplus is a highly-invasive tick that transmits the cattle parasites (Babesia bovis and B. bigemina) that cause cattle fever. R. microplus and Babesia are endemic in Mexico and ticks persist in the United States inside a narrow tick eradication quarantine area (TEQA) along the Rio Grande. This containment area is threatened by unregulated movements of illegal cattle and wildlife like white-tailed deer (WTD; Odocoileus virginianus). Methods Using 11 microsatellite loci we genotyped 1,247 R. microplus from 63 Texas collections, including outbreak infestations from outside the TEQA. We used population genetic analyses to test hypotheses about ecological persistence, tick movement, and impacts of the eradication program in southern Texas. We tested acaricide resistance with larval packet tests (LPTs) on 47 collections. Results LPTs revealed acaricide resistance in 15/47 collections (32%); 11 were outside the TEQA and three were resistant to multiple acaricides. Some collections highly resistant to permethrin were found on cattle and WTD. Analysis of genetic differentiation over time at seven properties revealed local gene pools with very low levels of differentiation (FST 0.00-0.05), indicating persistence over timespans of up to 29 months. However, in one neighborhood differentiation varied greatly over a 12-month period (FST 0.03-0.13), suggesting recurring immigration from distinct sources as another persistence mechanism. Ticks collected from cattle and WTD at the same location are not differentiated (FST = 0), implicating ticks from WTD as a source of ticks on cattle (and vice versa) and emphasizing the importance of WTD to tick control strategies. We identified four major genetic groups (K = 4) using Bayesian population assignment, suggesting multiple introductions to Texas. Conclusions Two dispersal mechanisms give rise to new tick infestations: 1) frequent short-distance dispersal from the TEQA; and 2) rare long

  17. Widespread movement of invasive cattle fever ticks (Rhipicephalus microplus) in southern Texas leads to shared local infestations on cattle and deer.

    PubMed

    Busch, Joseph D; Stone, Nathan E; Nottingham, Roxanne; Araya-Anchetta, Ana; Lewis, Jillian; Hochhalter, Christian; Giles, John R; Gruendike, Jeffrey; Freeman, Jeanne; Buckmeier, Greta; Bodine, Deanna; Duhaime, Roberta; Miller, Robert J; Davey, Ronald B; Olafson, Pia U; Scoles, Glen A; Wagner, David M

    2014-04-17

    Rhipicephalus (Boophilus) microplus is a highly-invasive tick that transmits the cattle parasites (Babesia bovis and B. bigemina) that cause cattle fever. R. microplus and Babesia are endemic in Mexico and ticks persist in the United States inside a narrow tick eradication quarantine area (TEQA) along the Rio Grande. This containment area is threatened by unregulated movements of illegal cattle and wildlife like white-tailed deer (WTD; Odocoileus virginianus). Using 11 microsatellite loci we genotyped 1,247 R. microplus from 63 Texas collections, including outbreak infestations from outside the TEQA. We used population genetic analyses to test hypotheses about ecological persistence, tick movement, and impacts of the eradication program in southern Texas. We tested acaricide resistance with larval packet tests (LPTs) on 47 collections. LPTs revealed acaricide resistance in 15/47 collections (32%); 11 were outside the TEQA and three were resistant to multiple acaricides. Some collections highly resistant to permethrin were found on cattle and WTD. Analysis of genetic differentiation over time at seven properties revealed local gene pools with very low levels of differentiation (FST 0.00-0.05), indicating persistence over timespans of up to 29 months. However, in one neighborhood differentiation varied greatly over a 12-month period (FST 0.03-0.13), suggesting recurring immigration from distinct sources as another persistence mechanism. Ticks collected from cattle and WTD at the same location are not differentiated (FST = 0), implicating ticks from WTD as a source of ticks on cattle (and vice versa) and emphasizing the importance of WTD to tick control strategies. We identified four major genetic groups (K = 4) using Bayesian population assignment, suggesting multiple introductions to Texas. Two dispersal mechanisms give rise to new tick infestations: 1) frequent short-distance dispersal from the TEQA; and 2) rare long-distance, human-mediated dispersal from

  18. Immunoprotection of cattle vaccinated with rRmLTI against Rhipicephalus microplus

    USDA-ARS?s Scientific Manuscript database

    Rhipicephalus microplus is arguably the most economically important cattle ectoparasite. There is interest in the development of second-generation anti-R. microplus vaccines with an improved product profile. Genes coding for trypsin inhibitors in R. microplus have diverse patterns of expression acro...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    DTIC Science & Technology

    2013-01-01

    insight into the sialome of the bedbug, Cimex lectularius, J. Proteome Res. 9 (2010) 3820–3831. [43] J. Ribeiro, T. Assumpcao, I. Francischetti, An...characterization of three cholinesterases from the common bed bug, Cimex lectularius, Insect. Mol. Biol. 21 (2012) 149–159. [45] I. Shaked, A. Meerson, Y. Wolf, R

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

    USDA-ARS?s Scientific Manuscript database

    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...

  2. 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.

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

    USDA-ARS?s Scientific Manuscript database

    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 (B...

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

    PubMed

    Miller, Robert; Estrada-Peña, Agustín; Almazán, Consuelo; Allen, Andrew; Jory, Lauren; Yeater, Kathleen; Messenger, Matthew; Ellis, Dee; Pérez de León, Adalberto A

    2012-08-17

    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 Rhipicephalus (Boophilus) microplus, through the efforts of the Cattle Fever Tick Eradication Program (CFTEP), with the exception of a permanent quarantine zone (PQZ) in south Texas along the border with Mexico. Keeping the U.S. free of cattle fever ticks in a sustainable manner is a critical national agricultural biosecurity issue. The efficacy of a Bm86-based anti-tick vaccine commercialized outside of the U.S. was evaluated against a strain of R. annulatus originated from an outbreak in Texas. Vaccination controlled 99.9 and 91.4% of the ticks 8 weeks and 5.5 months after the initial vaccination, respectively. Computer modeling of habitat suitability within the PQZ typically at risk of re-infestation with R. annulatus from Mexico predicted that at a level of control greater than 40%, eradication would be maintained indefinitely. Efficacy and computer modeling data indicate that the integration of vaccination using a Bm86-based anti-tick vaccine with standard eradication practices within the northwestern half of the PQZ could incentivize producers to maintain cattle on pasture thereby avoiding the need to vacate infested premises. Implementing this epidemiologically proactive strategy offers the opportunity to prevent R. annulatus outbreaks in the U.S., which would represent a significant shift in the way the CFTEP operates. Copyright © 2012. Published by Elsevier Ltd.

  5. 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

  6. 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

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

    USDA-ARS?s Scientific Manuscript database

    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...

  8. 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

  9. Assessment of bovine immunoprotecion against Rhipicephalus microplus using Bm86-CG antigen expressed in Pichia pastoris

    USDA-ARS?s Scientific Manuscript database

    The cattle tick Rhipicephalus microplus is a major pest of cattle in tropical and subtropical parts of the world. In addition to direct effects associated with its obligate parasitic way of life, R. microplus also transmits the pathogens that cause bovine babesiosis and anaplasmosis. Commercially ...

  10. 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.

  11. 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.

  12. 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.

  13. 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. Crown Copyright © 2016. Published by Elsevier Inc. All rights reserved.

  14. 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.

  15. First record of the pantropical blue tick Rhipicephalus microplus in Namibia.

    PubMed

    Nyangiwe, Nkululeko; Matthee, Conrad; Horak, Ivan; Matthee, Sonja

    2013-12-01

    The invasive pantropical blue tick, Rhipicephalus microplus, has recently been collected from cattle in Namibia. A cross-sectional study aimed at recording the geographic distribution of Rhipicephalus decoloratus and establishing whether R. microplus is present in Namibia was conducted towards the end of summer (March-April) 2013. Ticks were collected from cattle on 18 privately owned farms across a large geographical scale. Ticks were collected from three to five cattle per farm and species belonging to the genera Hyalomma and Rhipicephalus were recovered. Rhipicephalus decoloratus was present on all farms and R. microplus was recorded on four of the farms. The small numbers of R. microplus compared to R. decoloratus collected in the mixed infestations, suggests that the introduction events were recent.

  16. High efficacy of a 20 amino acid peptide of the acidic ribosomal protein P0 against the cattle tick, Rhipicephalus microplus.

    PubMed

    Rodríguez-Mallon, Alina; Encinosa, Pedro E; Méndez-Pérez, Lídice; Bello, Yamil; Rodríguez Fernández, Rafmary; Garay, Hilda; Cabrales, Ania; Méndez, Luis; Borroto, Carlos; Estrada, Mario Pablo

    2015-06-01

    Current strategies to control cattle ticks use integrated control programs (ICP) that include vaccination. Reduction in the use of chemicals and in the cost of tick control, the delay or elimination of acaricide resistance and the decreasing of environmental pollution are the advantages of using these programs. This integrated program is potentially applicable to all genotypes of chemical resistant ticks. However, the problem here is to improve the efficacy of anti-tick vaccines. The P0 protein is a structural component of the ribosome of all organisms. We have identified an immunogenic region of ribosomal protein P0 from Rhipicephalus spp. ticks that is not very conserved compared to the orthologous protein in their hosts. A synthetic 20 amino acid peptide from this sequence was effective as a vaccine against Rhipicephalus sanguineus infestations in an immunization and challenge experiment using rabbits. In this paper, the same peptide used as vaccine against the cattle tick Rhipicephalus Boophilus microplus shows a significant diminution in the number of engorged females recovered, in the weight of females and the weight of egg masses. The number of eggs hatched was also significantly reduced for the vaccinated group, with an overall effectivity for the antigen pP0 of 96%. These results, together with the conserved sequence of the P0 peptide among ticks, suggest that this antigen could be a good broad spectrum vaccine candidate. It would be expected to be active against many species of ticks and thus has promise in an ICP for effective control of ticks and thereby to improve the efficiency and productivity of the livestock industry. Copyright © 2015 Elsevier GmbH. All rights reserved.

  17. Acaricidal efficacy of Essentria® IC-3 and its active ingredients against acaricide resistant and susceptible strains of Rhipicephalus (Boophilus) microplus

    USDA-ARS?s Scientific Manuscript database

    Ticks and tick-borne diseases are a major constraint for a sustainable cattle industry in tropical and subtropical regions of the world. The development of resistance to most of the commonly used acaricides leads to an attempt to screen herbal products and their combinations for their possible acari...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  20. Therapeutic and residual efficacy of a pour-on formulation of Novaluron against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) on infested cattle

    USDA-ARS?s Scientific Manuscript database

    The effectiveness of a pour-on formulation of novaluron (Novatack Pour-on 5% AI), a benzoylphenyl urea acaricide, was evaluated by applying low and high rates (1 mg/20 kg body weight and 1 mg/10 kg body weight) to cattle infested with all parasitic developmental stages (adults, nymphs, and larvae) o...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  2. 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.

  3. 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.

  4. Cellular responses to Rhipicephalus microplus infestations in pre-sensitised cattle with differing phenotypes of infestation.

    PubMed

    Marufu, Munyaradzi C; Dzama, Kennedy; Chimonyo, Michael

    2014-02-01

    The blue tick, Rhipicephalus microplus, threatens cattle production in most tropical and subtropical areas of the world. Delayed skin hypersensitivity reactions are thought to cause Nguni cattle to be more resistant to R. microplus than Bonsmara cattle yet the cellular mechanisms responsible for these differences have not been classified. Tick counts and inflammatory cell infiltrates in skin biopsies from feeding sites of adult R. microplus ticks were determined in 9-month-old Nguni and Bonsmara heifers to determine the cellular mechanisms responsible for tick immunity. Nguni heifers (1.7 ± 0.03) had lower (P < 0.05) tick counts than the Bonsmaras (2.0 ± 0.03). Parasitized sites in Nguni heifers had higher counts of basophils, mast and mononuclear cells than those in the Bonsmara heifers. Conversely, parasitized sites in Nguni heifers had lower neutrophil and eosinophil counts than those in the Bonsmara heifers. Tick count was negatively correlated with basophil and mast cell counts and positively correlated with eosinophil counts in both breeds. In the Bonsmara breed, tick count was positively correlated with mononuclear cell counts. Cellular responses to adult R. microplus infestations were different and correlated with differences in tick resistance in Nguni and Bonsmara cattle breeds. It is essential to further characterise the molecular composition of the inflammatory infiltrate elicited by adult R. microplus infestation to fully comprehend immunity to ticks in cattle.

  5. 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.

  6. Designing and modeling of complex DNA vaccine based on tropomyosin protein of Boophilus genus tick.

    PubMed

    Ranjbar, Mohamamd Mahdi; Gupta, Shishir K; Ghorban, Khodayar; Nabian, Sedigheh; Sazmand, Alireza; Taheri, Mohammad; Esfandyari, Sahar; Taheri, Maryam

    2015-01-01

    Boophilus tick is a bloodsucking ectoparasite that transfers some pathogens, reducing production and thus leading to economical losses in the cattle industry. Tropomyosin (TPM) protein is a salivary protein, has actin regulator activity, and plays an important role in immune reactions against parasites. In the current study, besides developing a safe, effective, and broad spectrum protective measure against Boophilus genus tick based on TPM protein, we attempted to minimize possible problems occurring in the design of polytopic vaccines. Briefly, the steps that were followed in the present study were as follows: retrieving sequences and finding the mutational/conservative regions, selecting consensus and high immunogenic epitopes of B and CD4(+) T cells by different approaches, three-dimensional structure (3D structure) prediction and representation of epitopes and highly variable/conserve regions, designing vaccinal construct by fusion of B and T cell epitopes by special patterns and improving immunogenicity, evaluation of the constructs' primary structure and posttranslational modification, calculation of hydrophobic regions, reverse translation, codon optimization, open reading frame checking, insertion of start/end codon, Kozak sequence, and finally constructing the DNA vaccine. Variation plot showed some shared epitopes among the ticks' and mites' species that some might be effective only in some species. Finally, by following the steps mentioned above, two constructs for B and T cells were achieved. Checking constructs revealed their reliability and efficacy for in vitro production and utilization. Successful in silico modeling is an essential step of designing vigorous vaccines. We developed a novel protective and therapeutic vaccine against Boophilus genus (based on TPM protein). At the next step, constructed DNA vaccine would be produced in vitro and administrated to cattle, and its potency to induction of immune response and protection against Boophilus

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

    USDA-ARS?s Scientific Manuscript database

    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...

  8. 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. Copyright © 2016 Elsevier GmbH. All rights reserved.

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

  10. The transcription factor relish controls anaplasma marginale infection in the bovine tick rhipicephalus microplus

    USDA-ARS?s Scientific Manuscript database

    Rhipicephalus microplus is an important biological vector of Anaplasma marginale, the etiological agent of bovine anaplasmosis. The knowledge of tick immune responses to control bacterial infections remains limited. In this study, we demonstrate that transcription factor Relish from the Imd signalin...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  12. First report of the cattle tick Rhipicephalus microplus resistant to ivermectin in Mexico

    USDA-ARS?s Scientific Manuscript database

    Three cattle farms with ticks, Rhipicephalus microplus, thought to be resistant to ivermectin in Yucatan, Mexico, were studied (SFDO, SPN, LUADY). Each field population was collected and tested twice, several months apart. The larval immersion test was used on the progeny of collected adult females ...

  13. Selection of an ivermectin-resistant strain of Rhipicephalus microplus (Acari: Ixodidae) in Brazil

    USDA-ARS?s Scientific Manuscript database

    Resistance to ivermectin (IVM) in field populations of Rhipicephalus microplus of Brazil has been observed since 2001. In this work, four selection methods (infestations with: (1) IVM-treated larvae; (2) larvae from IVM-treated adult female ticks; (3) larvae from IVM-treated adult female ticks on an...

  14. 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.

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

    USDA-ARS?s Scientific Manuscript database

    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.

    USDA-ARS?s Scientific Manuscript database

    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. Protective Immunity Against Tick Infestation in Cattle Vaccinated with Recombinant Trypsin Inhibitor of Rhipicephalus Microplus

    USDA-ARS?s Scientific Manuscript database

    The southern cattle fever tick, Rhipicephalus microplus, and the related cattle fever tick species, R. annulatus, collectively referred to herein as Cattle Fever Ticks (CFT), transmit Babesia bovis and B. bigemina, and Anaplasma marginale, that cause bovine babesiosis and anaplasmosis, respectively....

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

  19. 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

  20. In vitro acaricidal activity of Crescentia cujete L. fruit pulp against Rhipicephalus microplus.

    PubMed

    Pereira, Sinval Garcia; de Araújo, Sandra Alves; Guilhon, Giselle Maria Skelding Pinheiro; Santos, Lourivaldo Silva; Junior, Livio Martins Costa

    2017-05-01

    Cattle tick (Rhipicephalus microplus) is an important constraint on livestock production, particularly in tropical and subtropical areas. Synthetic acaricides are the primary method of control this thick. Resistance of R. microplus to synthetic acaricides has given rise to the need of new scientific investigations on alternative ways to control it. The objective of this work was to conduct a chemical investigation of Crescentia cujete L. fruit pulp and to evaluate the acaricide effect on R. microplus larvae of extracts, fractions, and isolated substances from this species. Crescentia cujete is a Bignoniaceae and it is characteristic of tropical climates. Its most distinctive feature is its fruit, which is spherical and gourd-like, with a hard woody shell and gelatinous pulp. The fruit pulp of C. cujete was extracted with ethanol by maceration yielding an extract and a residue. The extract was partitioned giving an ethyl acetate phase (EAF) that was fractionated by chromatographic procedures yielding cinnamic acid and benzoic acid. The remained botanic material from the ethanol extraction was further and successively extracted with ethyl ether and methanol under reflux. The acaricide effect of all extracts and fractions was evaluated against R. microplus using the larval packet test. The best result was observed for the EAF that caused 100% of mortality when a 10% solution was used and with LC50 of 5.9%. Cinnamic acid was also tested showing 63.0% of mortality with LC50 of 6.6% at the same concentrations and time of the experiment. The data obtained in this study provides information related to the chemical nature and the acaricide activity of the C. cujete epicarp that has not been previously reported. This study shows that the ethyl acetate phase of the ethanol extract of C. cujete fruit pulp is a potential alternative control for R. microplus and its activity is in part associated to cinnamic acid, its major compound.

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

    PubMed

    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

  2. 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.

  3. 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.

  4. SNPs and other polymorhisms associated with acaricide resistance in Rhipicephalus microplus.

    PubMed

    Aguilar, Gabriela; Olvera, Andrea Margarita; Carvajal, Bertha Isabel; Mosqueda, Juan

    2018-01-01

    Ixodicides resistance of ticks is one of the most important problems for the livestock industry in tropical and subtropical regions, mainly due to the increase in cases of multiple resistance in all families of the ixodicides used. Molecular markers such as single nucleotide polymorphisms (SNPs) has been proposed to identify the resistance to ixodicides in Rhipicephalus microplus. Many studies have recently been conducted using SNPs and other types of molecular markers to determine if they are associated with resistance to different products in many parts of the world. Knowing these changes at the molecular level, will allow to establish mechanisms and control strategies for the use of ixodicides. In this review, we will discuss and describe the different SNPs and other polymorphisms associated with resistance in R. microplus.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  6. 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.

  7. Entomopathogenic nematodes in insect cadaver formulations for the control of Rhipicephalus microplus (Acari: Ixodidae).

    PubMed

    Monteiro, Caio Márcio de Oliveira; Matos, Renata da Silva; Araújo, Laryssa Xavier; Campos, Roberson; Bittencourt, Vânia Rita Elias Pinheiro; Dolinski, Claudia; Furlong, John; Prata, Márcia Cristina de Azevedo

    2014-07-14

    This study evaluated the efficacy of four entomopathogenic nematode (EPN) strains in insect cadaver formulations against Rhipicephalus microplus and compared the efficacy of the most virulent EPNs applied in cadavers of Galleria mellonella and Tenebrio molitor. In the first experiment, infected G. mellonela larvae were used as the source of EPNs. Engorged females of R. microplus were placed in pots filled with soil and different numbers of G. mellonella larvae infected with one of four species of nematodes. All treatments with EPNs of the genus Heterorhabditis caused significant reduction (p<0.05) in the egg mass weight and hatching percentage of larvae. The EPNs of the genus Steinernema, except for the group exposed to Steinernema carpocapsae ALL, whose source nematodes included six larvae of G. mellonella, caused a significant reduction (p<0.05) in the egg mass weight produced per female. Steinernema feltiae SN applied with two, four, and six cadavers and S. carpocapsae ALL with two cadavers caused a reduction in hatching percentage of larvae of R. microplus (p<0.05). The percentage of control was above 95% in all groups treated with Heterorhabditis bacteriophora HP88 and Heterorhabditis indica LPP1 and in the treatment with four larvae infected with S. feltiae SN. The second experiment followed the same methodology, using G. mellonella and T. molitor larvae infected by the two most virulent EPNs. H. bacteriophora HP88 and H. indica LPP1 in different formulations caused reduction in the egg mass weight and hatching percentage of larvae. The percentage of control were 82.4 and 84.9% for H. bacteriophora HP88 and H. indica LPP1, respectively, formulated in T. molitor, and reaching 99.9% in groups formulated with G. mellonella. The EPNs tested in insect cadaver formulation showed pathogenicity to engorged females of R. microplus and EPNs of the genus Heterorhabditis formulated in G. mellonella larvae were more effective. Copyright © 2014 Elsevier B.V. All rights

  8. An integrative approach to understanding pyrethroid resistance in Rhipicephalus microplus and R. decoloratus ticks.

    PubMed

    Wyk, Roelof Dj van; Baron, Samantha; Maritz-Olivier, Christine

    2016-06-01

    Rhipicephalus microplus and Rhipicephalus decoloratus species occur in regions with savannah and temperate climates, typically in grassland and wooded areas used as cattle pasture. Both species are associated with the transmission of Anaplasma and Babesia spp., impacting livestock health and quality of livestock-associated products. In Africa, tick control is predominantly mediated with the use of acaricides, such as synthetic pyrethroids. After several years on the market, reports of resistance to synthetic pyrethroids escalated but limited field data and validation studies have been conducted to determine the extent of acaricide resistance in Africa. Without this data, knowledge-based tick control will remain problematic and selection pressure will remain high increasing the rate of resistance acquisition. To date, several pyrethroid resistance associated single nucleotide polymorphisms (SNPs) have been reported for arthropods within the voltage-gated sodium channel. Three SNPs have been identified within this channel in pyrethroid resistant R. microplus ticks, but none has been reported for R. decoloratus. This study is the first to report the presence of a shared SNP within the voltage-gated sodium channel in both R. microplus and R. decoloratus, which is directly linked to pyrethroid resistance in R. microplus. As the mode of action by which these SNPs mediate pyrethroid resistance remains unknown, this study aims to set hypotheses by means of predictive structural modelling. This not only paves the way forward to elucidating the underlying biological mechanisms involved in pyrethroid resistance, but also improvement of existing acaricides and ultimately sustainable tick control management. Copyright © 2016 Elsevier GmbH. All rights reserved.

  9. Protective immunity against tick infestation in cattle vaccinated with recombinant trypsin inhibitor of Rhipicephalus microplus.

    PubMed

    Andreotti, Renato; Cunha, Rodrigo Casquero; Soares, Mariana Aparecida; Guerrero, Felix D; Leite, Fábio P Leivas; de León, Adalberto A Pérez

    2012-10-19

    The cattle tick, Rhipicephalus microplus, is regarded as the most economically important ectoparasite of livestock globally. Control is achieved primarily through the use of acaricides. This approach is hampered by the development of resistance to commercial acaricides among cattle tick populations. Vaccination against R. microplus infestation is another technology that can be integrated for effective cattle tick control. Proteins belonging to the Kunitz-BPTI family are abundant in cattle tick salivary glands, midgut, and ovaries. These organs are attractive targets for the development of a novel cattle tick vaccine. Efficacy assessment against cattle tick infestation in bovines using a vaccine containing the recombinant form of a member of the Kunitz family from R. microplus produced in a yeast expression system is reported for the first time here. The yeast Pichia pastoris was bioengineered to produce the recombinant version of a trypsin inhibitor that is expressed in cattle tick larvae (rRmLTI). Immunization with rRmLTI afforded 32% efficacy against R. microplus. The estimated molecular weight of rRmLTI was 46 kDa. Structural homology to the native form of the larval trypsin inhibitor was documented by recognition of rRmLTI in Western-blots using polyclonal antibodies from mice immunized with cattle tick larval extract or rRmLTI. Bioinformatics analysis of the partial nucleotide and deduced amino acid sequences indicated that the rRmLTI closely resembles BmTI-6, which is a three-headed Kunitz protein present in cattle tick ovary and fat tissue. Published by Elsevier Ltd.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. Immunoregulation of bovine macrophages by factors in the salivary glands of Rhipicephalus microplus

    PubMed Central

    2012-01-01

    Background Alternative strategies are required to control the southern cattle tick, Rhipicephalus microplus, due to evolving resistance to commercially available acaricides. This invasive ectoparasite is a vector of economically important diseases of cattle such as bovine babesiosis and anaplasmosis. An understanding of the biological intricacies underlying vector-host-pathogen interactions is required to innovate sustainable tick management strategies that can ultimately mitigate the impact of animal and zoonotic tick-borne diseases. Tick saliva contains molecules evolved to impair host innate and adaptive immune responses, which facilitates blood feeding and pathogen transmission. Antigen presenting cells are central to the development of robust T cell responses including Th1 and Th2 determination. In this study we examined changes in co-stimulatory molecule expression and cytokine response of bovine macrophages exposed to salivary gland extracts (SGE) obtained from 2-3 day fed, pathogen-free adult R. microplus. Methods Peripheral blood-derived macrophages were treated for 1 hr with 1, 5, or 10 μg/mL of SGE followed by 1, 6, 24 hr of 1 μg/mL of lipopolysaccharide (LPS). Real-time PCR and cytokine ELISA were used to measure changes in co-stimulatory molecule expression and cytokine response. Results Changes were observed in co-stimulatory molecule expression of bovine macrophages in response to R. microplus SGE exposure. After 6 hrs, CD86, but not CD80, was preferentially up-regulated on bovine macrophages when treated with 1 μg/ml SGE and then LPS, but not SGE alone. At 24 hrs CD80, CD86, and CD69 expression was increased with LPS, but was inhibited by the addition of SGE. SGE also inhibited LPS induced upregulation of TNFα, IFNγ and IL-12 cytokines, but did not alter IL-4 or CD40 mRNA expression. Conclusions Molecules from the salivary glands of adult R. microplus showed bimodal concentration-, and time-dependent effects on differential up-regulation of CD

  16. Molecular characterisation of the tick Rhipicephalus microplus in Malaysia: new insights into the cryptic diversity and distinct genetic assemblages throughout the world.

    PubMed

    Low, Van Lun; Tay, Sun Tee; Kho, Kai Ling; Koh, Fui Xian; Tan, Tiong Kai; Lim, Yvonne Ai Lian; Ong, Bee Lee; Panchadcharam, Chandrawathani; Norma-Rashid, Yusoff; Sofian-Azirun, Mohd

    2015-06-24

    The morphotaxonomy of Rhipicephalus microplus complex has been challenged in the last few years and prompted many biologists to adopt a DNA-based method for distinguishing the members of this group. In the present study, we used a mitochondrial DNA analysis to characterise the genetic assemblages, population structure and dispersal pattern of R. microplus from Southeast Asia, the region where the species originated. A phylogeographic analysis inferred from the 16S rRNA and cytochrome oxidase subunit I (COI) genes was performed with five populations of R. microplus collected from cattle in Malaysia. Malaysian R. microplus sequences were compared with existing COI and 16S rRNA haplotypes reported globally in NCBI GenBank. A total of seven and 12 unique haplotypes were recovered by the 16S rRNA and COI genes, respectively. The concatenated sequences of both 16S rRNA and COI revealed 18 haplotypes. Haplotype network and phylogenetic analyses based on COI+16S rRNA sequences revealed four genetically divergent groups among Malaysian R. microplus. The significantly low genetic differentiation and high gene flow among Malaysian R. microplus populations supports the occurrence of genetic admixture. In a broader context, the 16S rRNA phylogenetic tree assigned all isolates of Malaysian R. microplus into the previously described African/the Americas assemblage. However, the COI phylogenetic tree provides higher resolution of R. microplus with the identification of three main assemblages: clade A sensu Burger et al. (2014) comprises ticks from Southeast Asia, the Americas and China; clade B sensu Burger et al. (2014) is restricted to ticks that originated from China; and clade C sensu Low et al. (2015) is a new genetic assemblage discovered in this study comprising ticks from India and Malaysia. We conclude that the R. microplus complex consisting of at least five taxa: R. australis, R. annulatus, R. microplus clade A sensu Burger et al. (2014), R. microplus clade B sensu

  17. 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.

  18. 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.

  19. Cellular response in the tick feeding site in crossbred cattle artificially infested by Rhipicephalus microplus.

    PubMed

    Engracia Filho, Jair Rodini; Araújo, Chiara Domingues; Pinto, Gabriela Nishihara; Mendes, Yann Henrique; Bechara, Gervasio Henrique

    2017-06-01

    Ticks and tick-borne diseases are of global importance and can cause serious economic losses to cattle-raising businesses. However, only few attempts have been made to describe the relationships between various cellular immune components and tick counts of, and therefore resistance to, Rhipicephalus microplus. The present study evaluated the late inflammatory cellular response and dermal dendritic cells at the attachment site in naturally presensitized bovines, artificially infested by R. microplus ticks. Twenty-two crossbred (Holstein × Gir) male experimental cattle were artificially infested with approximately 10,000 larvae of R. microplus. Parasitic infestation was determined by counting the engorged female ticks (4.5-8.0 mm in diameter) at the 21st day after artificial infestation. Biopsies (6 mm) were taken prior to the infestation and along with the tick count from the right pinna of each animal, and examined with histological techniques and immunohistochemistry using S100 protein. Inflammatory cell counts were conducted in the sections stained with the May-Grünwald Giemsa technique, and immunostained dermal dendritic cells were evaluated and classified in scores ranging from 0 to 4. An average of 39.2 ticks per animal was found. Eosinophils (47.8%) constituted the major portion of the cellular infiltrate, followed by mononuclear cells (28.3%), neutrophils (14.4%), and basophils (0% to approximately 2%). The dendritic cell count shows a considerable population in the dermis, with pre- and post-infestation mean scores of 1.54 and 1.89, respectively; these scores were not significantly different. Our results pointed out the importance of the cellular response in the cattle resistance to ticks.

  20. Does Rhipicephalus microplus tick infestation increase the risk for myiasis caused by Cochliomyia hominivorax in cattle?

    PubMed

    Reck, José; Marks, Fernanda S; Rodrigues, Rogério O; Souza, Ugo A; Webster, Anelise; Leite, Romário C; Gonzales, João Carlos; Klafke, Guilherme M; Martins, João Ricardo

    2014-01-01

    The larval phase of Cochliomyia hominivorax (screwworm) is an obligate parasite of vertebrate animals, particularly mammals, and widespread in South America, where it remains one of the most important parasitic diseases of domestic animals. The skin of cattle highly infested by ticks, with cutaneous lesions, exudation of tissue fluid and blood scent seems to produce the ideal environment for fly attraction. However, an association between these parasites was never investigated. The aim of this work was to verify if there is an association between Rhipicephalus microplus tick load and the occurrence of C. hominivorax myiasis in cattle, and to quantify the risk. Sixty bovine (Bos taurus taurus, Angus breed) under field conditions were observed for 24 weeks, during which weekly tick counts and examination for the presence of myiasis were performed. There was a significant association between a high tick burden (24-week mean above 50 ticks per animal) and myiasis occurrence (P=0.0102). The calculated relative risk (RR) for C. hominivorax myiasis occurrence in cattle with high tick burden was 3.85 (CI95%=1.23-12.13); indicating that cattle highly parasitized by R. microplus have about four times more risk of myiasis than those with a low parasite load. As far as we aware, this is the first statistically based evidence of the relationship between R. microplus parasitic load and occurrence of myiasis by C. hominivorax. This result could be useful for the design of integrated control strategies for these parasites and to provide more information for the understanding of cattle tick parasitism in cattle production. Copyright © 2013 Elsevier B.V. All rights reserved.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  4. First report of a Rhipicephalus microplus tick population multi-resistant to acaricides and ivermectin in the Mexican tropics.

    PubMed

    Fernández-Salas, A; Rodríguez-Vivas, R I; Alonso-Díaz, M A

    2012-02-10

    We document the presence of a Rhipicephalus microplus tick population resistant to acaricides (organophosphates (OP), synthetic pyrethroids (SP), amitraz) and macrocyclic lactones (ML) (ivermectin). Engorged females of R. microplus were collected from a cattle farm in Veracruz, Mexico, to evaluate acaricide and ivermectin resistance. The modified larval packet test (LPT) was used to detect OP (chlorpiriphos and diazinon) and SP (flumethrin, deltamethrin and cypermethrin) resistance and the larval immersion test (LIT) to detect resistance to amitraz and ivermectin. Both, LPT and LIT were performed twice at different times with different collected samples. Mortality data with ivermectin were subjected to probit analysis to obtain lethal concentrations and resistance ratios (RR) using an ivermectin-susceptible strain (Deutch) as a reference. The R. microplus population showed resistance to all acaricides tested, with different mortalities at the discriminate dose: chlorpiriphos (1%), diazinon (24.2%), flumethrin (92.8%), deltamethrin (94.2%), cypermethrin (98.0%) and amitraz (1.5%). The studied tick population also showed resistance to ivermectin with a resistance ratio at 99% of 9.58 and 6.52 in the first and second evaluation, respectively. We report for the first time a R. microplus population in Mexico with different levels of resistance to OP, SP, amidines (Am) and ivermectin. The uncontrolled use of these products in the study area may promote the complete failure of tick control within a short period of time. Copyright © 2011 Elsevier B.V. All rights reserved.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  7. 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.

  8. 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.

  9. Chemical composition and acaricide activity of an essential oil from a rare chemotype of Cinnamomum verum Presl on Rhipicephalus microplus (Acari: Ixodidae).

    PubMed

    Monteiro, Ildenice Nogueira; Monteiro, Odair Dos Santos; Costa-Junior, Lívio Martins; da Silva Lima, Aldilene; Andrade, Eloisa Helena de Aguiar; Maia, José Guilherme Soares; Mouchrek Filho, Victor Elias

    2017-04-30

    The Essential Oils (EOs) from the leaves of species Cinnamomum verum J. Presl are used in the pharmaceutical industry for their numerous biological activities. Currently, the main compound of C. verum EO is eugenol which has acaricidal activity; however, a rare chemotype with benzyl benzoate as the main component can be found. Benzyl benzoate is recognized as an acaricide; however, studies of the C. verum EOs benzyl benzoate chemotype on Rhipicephalus microplus were not reported. The aim of this study was to evaluate the acaricide activity of an EO from a rare chemotype of C. verum, as well as purified benzyl benzoate, against larvae and engorged females of R. microplus resistant to amidines and pyrethroids. The EO was extracted from C. verum leaves and the compounds present were identified using a gas phase chromatograph coupled to a mass spectrometer. Efficacy against R. microplus was assessed by the larval packet and the engorged female immersion tests. A rare chemotype of C. verum was found to produce EOs with benzyl benzoate (65.4%) as the main compound. The C. verum essential oil was 3.3 times more efficient on the R. microplus larvae than was benzyl benzoate. However, no differences were found on the R. microplus engorged females. This is the first report regarding the acaricidal activity of C. verum with chemotype benzyl benzoate, and this compound showed acaricidal activity on R. microplus larvae. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. A model to simulate the effect of vaccination against Boophilus ticks on cattle.

    PubMed

    Lodos, J; Boue, O; de la Fuente, J

    2000-02-01

    This paper describes a vaccination model to simulate the effect of cattle vaccination with concealed antigens on Boophilus tick spp. The model considers the vaccination effect in three parts: antibody titer, accumulation of damaging vaccination effects by parasite stages, and the effect of accumulated damage on all tick life stages. Biological parameters for ticks and hosts, as well as parameters describing tick-host interaction, were included. The validity of this model, integrated with the TICKSIM simulation program, was demonstrated for the Bm86-containing vaccine Gavac by comparing simulated and real data for several geographic locations in the Americas. All model parameters were estimated using field data collected in the different geographic locations. The model sensitivity to changes in antibody titer level and titer half-life was studied, and the impact on tick population density of changes in these parameters was evaluated. Simulation results showed that to achieve a higher level of tick control, an increase in the maximum antibody titer levels was more important than extending titer half-life in geographical locations with short seasonal peaks of tick infestation. The TICKSIM program, integrated with the new vaccination model, proved to be a framework for designing and evaluating tick control strategies, including vaccination with GavacTM.

  11. 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

  12. 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.

  13. 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. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Activity of carboxylesterases, glutathione-S-transferase and monooxygenase on Rhipicephalus microplus exposed to fluazuron.

    PubMed

    Gaudêncio, Fabrício Nascimento; Klafke, Guilherme Marcondes; Tunholi-Alves, Vinícius Menezes; Ferreira, Thaís Paes; Coelho, Cristiane Nunes; da Fonseca, Adivaldo Henrique; da Costa Angelo, Isabele; Pinheiro, Jairo

    2017-10-01

    The objective of this study was to assess the effect of the exposure to fluazuron on the activity of common pesticide detoxification enzyme groups in the cattle tick (Rhipicephalus microplus). Engorged females of a susceptible strain (POA) and a resistant strain (Jaguar) were exposed in vitro to fluazuron and their eggs and larvae were used to compare the activities of the general esterases, mixed-function oxidases (MFO) and glutathione-S-transferase (GST). The results showed significant elevation in MFO contents and esterases activity in the resistant strain when compared with the susceptible strain, in eggs and larvae respectively. In the POA strain, the MFO activity in eggs was down-regulated by fluazuron exposure. Based on these results, it can be concluded that different detoxification enzymes can act in distinct pathways depending on the tick's development stage, and may be related to fluazuron detoxification in resistant strains. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Injectable fipronil for cattle: Plasma disposition and efficacy against Rhipicephalus microplus.

    PubMed

    Cid, Yara P; Ferreira, Thais P; Magalhães, Viviane S; Correia, Thais R; Scott, Fábio B

    2016-04-15

    Fipronil is a phenylpyrazole class insecticide. It is widely used as an insecticide in agriculture and in the control of ectoparasites in veterinary medicine. The application of fipronil in an injectable form (subcutaneously) becomes an innovation, since there is no commercially available preparation containing fipronil herein. The present study aimed at fipronil usage, applied subcutaneously in cattle, to control Rhipicephalus microplus. The assessing criteria used in the research have been the construction of the plasma concentration curve and efficacy studies. A method using High Performance Liquid Chromatograph with ultraviolet detection was developed for determination of fipronil in bovine plasma samples, providing a fast and simple process with good reproducibility and low limit of quantification. The validation of the analytical method showed linearity, selectivity, precision, accuracy, sensitivity and stability, thus proving it as suitable for routine analysis. This method showed to be an important investigative tool in the analysis of fipronil plasma concentration in cattle. Fipronil administered via subcutaneous in bovine reached the systemic circulation (Cmax=378.06±137.44 ng/mL), was quickly absorbed (t(max)=10±0.87 h), and its elimination occurred slowly (t(1/2)=12 days), while maintaining quantifiable blood plasma levels (23.79±12.16 ng/mL) for up to 21 days after the treatment with a 1 mg/kg dosage. The in vivo efficacy tests proved that fipronil applied subcutaneously in a single dose of 1 mg/kg in cattle exhibited a mean efficacy of 82.41% against R. microplus. The potential of subcutaneous injection as an alternative treatment route in cattle encourage the development of an injectable formulation of fipronil. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Entomopathogenic nematodes associated with essential oil of Lippia sidoides for control of Rhipicephalus microplus (Acari: Ixodidae).

    PubMed

    Monteiro, Caio Márcio Oliveira; Araújo, Laryssa Xavier; Gomes, Geovany Amorim; Senra, Tatiane Oliveira Souza; Calmon, Fernanda; Daemon, Erik; de Carvalho, Mario Geraldo; Bittencourt, Vânia Rita Elias Pinheiro; Furlong, John; Prata, Márcia Cristina de Azevedo

    2014-01-01

    In vitro effect of the association of the entomopathogenic nematodes Heterorhabditis bacteriophora HP88 and Heterorhabditis indica LPP1 with the essential oil of Lippia sidoides on engorged females of Rhipicephalus microplus was assessed. Engorged females with homogeneous weights (p >0.05) were divided into six groups of ten ticks each (each female = an experimental unit). In the treated groups, the ticks were exposed to the oil (40 μl/ml=4%) and nematodes (300 entomopathogenic nematodes (EPNs/tick) separately and also to the nematodes together with the oil, while in the control group, the females were immersed in Tween 80 (3%). All the treatments caused a significant reduction in the egg mass weight (p <0.05). The hatching percentage of larvae was reduced (p <0.05) in the groups treated with H. bacteriophora HP88, H. indica LPP1, and H. bacteriophora HP88 associated with the oil, but in the group treated only with the oil, the hatching of percentage of larvae rate was similar to that of the control group (p >0.05). The percentage of control in the group exposed only to the oil was 50.7%, while in the other treated groups, the percentage of control was greater than 95 %, reaching 100 %in the group treated with H. indica LPP1 associated with the oil. The results of the L. sidoides essential oil in the concentrations tested in association with both H. bacteriophora HP88 and H. indica LPP1 indicate that these combinations can be used to control R. microplus.

  17. Cytotoxic effects of extract of Acmella oleraceae (Jambú) in Rhipicephalus microplus females ticks.

    PubMed

    de Oliveira, Patrícia Rosa; de Carvalho Castro, Karina Neoob; Anholeto, Luis Adriano; Camargo Mathias, Maria Izabel

    2016-08-01

    The present study analyzed the effects of different concentrations of the hexane extract of A. oleraceae (HEAO) (Jambú) on the germ cells of semi-engorged Rhipicephalus microplus female ticks, through a morpho-histological study, evaluating the effectiveness of the extract in the genesis of the individuals. To perform this analysis, 100 semi-engorged females were divided into five groups with 20 individuals each: groups I and II, respectively constituted by distilled water control and 50% ethanol + 1% DMSO, and groups III, IV, and V constituted by treatment with HEAO in the concentrations of 12.5, 25.0 and 50.0 mg/mL, respectively. All the ticks were immersed in the different concentrations of the extract or in distilled water for 5 minutes, dried and conditioned in BOD incubator for 7 days. The individuals of the treatment groups revealed the action of this extract showing alterations in the germ cells of the females from the different groups when compared with those from the groups I and II (control groups). These alterations were mainly related to the size and shape of the oocytes; number of yolk granules; presence, number, size and location of vacuoles in the cytoplasm of all the germ cells; and the presence of nuclear alterations in these cells as well. Thus, it was demonstrated that the concentrations of HEAO affected the germ cells of R. microplus ticks. The effects of the extract are similar to those caused by renowned and efficient chemical products used to control these ticks. Microsc. Res. Tech. 79:744-753, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  18. 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.

  19. 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. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Toxicity of Neem Seed Oil against the Larvae of Boophilus decoloratus, A One-Host Tick In Cattle

    PubMed Central

    Choudhury, M. K.

    2009-01-01

    The in vitro toxicity of neem seed oil (Azadirachta indica A. Juss, family: Meliaceae, Dogon yaro in Hausa language in Nigeria) was tested against the larvae of a one-host tick, Boophilus decoloratus (family: Ixodidae or hard tick, commonly known as blue tick) parasitic mainly to cattle generally found in savannas of tropical equatorial Africa. The 20, 40, 60, 80 and 100% concentrations of neem seed oil were found to kill all (100% mortality) the larvae after 27, 27, 27, 27 and 24 h respectively. PMID:20502579

  1. Toxicity of Neem Seed Oil against the Larvae of Boophilus decoloratus, A One-Host Tick In Cattle.

    PubMed

    Choudhury, M K

    2009-09-01

    The in vitro toxicity of neem seed oil (Azadirachta indica A. Juss, family: Meliaceae, Dogon yaro in Hausa language in Nigeria) was tested against the larvae of a one-host tick, Boophilus decoloratus (family: Ixodidae or hard tick, commonly known as blue tick) parasitic mainly to cattle generally found in savannas of tropical equatorial Africa. The 20, 40, 60, 80 and 100% concentrations of neem seed oil were found to kill all (100% mortality) the larvae after 27, 27, 27, 27 and 24 h respectively.

  2. 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

    USDA-ARS?s Scientific Manuscript database

    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...

  3. 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.

  4. 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.

  5. 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.

  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. © 2016 The Royal Entomological Society.

  7. 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.

  8. 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.

  9. 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.

  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. 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

  15. 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

    USDA-ARS?s Scientific Manuscript database

    The aim of this study was to determine whether remote sensed data (Landsat Thematic Mapper imagery) could be used to identify white-tailed deer, Odocoileus virginianus (Zimmerman), and habitat and target locations for sampling free-living larvae of Southern Cattle Ticks, Rhipicephalus (Boophilus) mi...

  16. 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

  17. 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.

  18. Host resistance in cattle to infestation with the cattle tick Rhipicephalus microplus.

    PubMed

    Jonsson, N N; Piper, E K; Constantinoiu, C C

    2014-11-01

    Resistance to Rhipicephalus microplus infestation in cattle has many effector mechanisms, each of which is likely to be modulated by complex, interacting factors. Some of the mechanisms of host resistance and their modulating factors have been identified and quantified, although much remains to be explained. The variation in resistance to tick infestation is most marked between Bos taurus and Bos indicus cattle, taurine cattle given the same exposure carrying between five and 10 times as many ticks as indicine cattle. Tick resistance is mostly manifest against attaching larvae, which attempt to feed often and without success, death occurring mostly within 24 h of finding a host. There is evidence of innate and adaptive immune response to tick infestation, and it appears that the relative importance of each differs between indicine and taurine cattle. There is conflicting information regarding the role of humoral immunity in tick resistance, and recent studies indicate that strong IgG responses to tick antigens are not protective. A strong T-cell-mediated response directed against larval stages, as mounted by indicine cattle, seems to be protective. Variation in the extracellular matrix of skin (epidermal growth factors, collagens and other matrix components such as lumican) also contributes to variation in host resistance. © 2014 John Wiley & Sons Ltd.

  19. In vitro effect of seven essential oils on the reproduction of the cattle tick Rhipicephalus microplus.

    PubMed

    Pazinato, Rafael; Volpato, Andréia; Baldissera, Matheus D; Santos, Roberto C V; Baretta, Dilmar; Vaucher, Rodrigo A; Giongo, Janice L; Boligon, Aline A; Stefani, Lenita Moura; Da Silva, Aleksandro Schafer

    2016-11-01

    The acaricidal effect of seven essential oils was examined in vitro against the cattle tick (Rhipicephalus microplus). Engorged female ticks were manually collected in farms of Southern Brazil and placed into petri dishes (n = 10) in order to test the following oils: juniper (Juniperus communis), palmarosa (Cymbopogon martinii), cedar (Cedrus atlantica), lemon grass (Cymbopogon citratus), ginger (Zingiber officinale), geranium (Pelargonium graveolens) and bergamot (Citrus aurantium var bergamia) at concentrations of 1%, 5%, and 10% each. A control group was used to validate the tests containing Triton X-100 only. Treatment effectiveness was measured considering inhibition of tick oviposition (partial or total), egg's weight, and hatchability. C. martinii, C. citratus and C. atlantica essential oils showed efficacy higher than 99% at all concentrations tested. In addition, J. communis, Z. officinale, P. graveolens, and C. aurantium var bergamia oils showed efficiency ranging from 73% to 95%, depending on the concentration tested, where higher concentrations showed greater efficacy. It was concluded that essential oils can affect tick reproduction in vitro by inhibiting oviposition and hatchability.

  20. 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.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  4. Gene-enriched draft genome of the cattle tick Rhipicephalus microplus: Assembly by the hybrid Pacific Biosciences/Illumina approach enabled analysis of the highly repetitive genome

    USDA-ARS?s Scientific Manuscript database

    The genome of the cattle tick R. microplus, an ectoparasite with global distribution, is estimated to be 7.1 Gbp and consists of ~70% repetitive DNA. We report the first assembly of a tick genome that utilized a hybrid sequencing and assembly approach to capture the repetitive fractions of the genom...

  5. 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

  6. 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

  7. Effect of new ethyl and methyl carbamates on Rhipicephalus microplus larvae and adult ticks resistant to conventional ixodicides.

    PubMed

    Pérez-González, I E; Prado-Ochoa, M G; Muñoz-Guzmán, M A; Vázquez-Valadez, V H; Velázquez-Sánchez, A M; Avila-Suárez, B L; Cuenca-Verde, C; Angeles, E; Alba-Hurtado, F

    2014-01-31

    The effects of six new synthetic carbamates on fully engorged females of four Rhipicephalus microplus strains (one reference strain susceptible to conventional ixodicides, two strains multiresistant to ixodicides and one tick field isolate) were compared. In addition, the effect of two other new synthetic carbamates was tested on larvae from the same strains. The first six tested carbamates reduced egg laying and inhibited egg hatching in the four studied strains (P<0.05). Compared with untreated females, the eggs produced by the treated engorged female ticks of all strains had a dark, dry, opaque appearance and were less adherent. The remaining two tested carbamates induced larval mortality in all of the evaluated strains. The three studied R. microplus strains displayed 50% resistance ratios (RR50) of less than 2 when compared to the susceptible reference strain. These results demonstrate that both carbamates with a larvicidal effect and carbamates that inhibit egg laying and embryo development are efficacious against tick strains that are resistant to commercial ixodicides, no cross resistance was observed. Copyright © 2013 Elsevier B.V. All rights reserved.

  8. 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.

  9. Nanostructured cinnamon oil has the potential to control Rhipicephalus microplus ticks on cattle.

    PubMed

    Dos Santos, Daiane S; Boito, Jhonatan P; Santos, Roberto C V; Quatrin, Priscilla M; Ourique, Aline Ferreira; Dos Reis, João H; Gebert, Roger R; Glombowsky, Patrícia; Klauck, Vanderlei; Boligon, Aline A; Baldissera, Matheus D; Da Silva, Aleksandro S

    2017-08-29

    The aim of this study was to evaluate the capacity of pure and nanostructured cinnamon oil to control the infestation and reproductive efficiency of Rhipicephalus microplus on dairy cows. In vitro (stage I)-engorged female ticks were immersed in concentrations of 1.0, 5.0 and 10% of cinnamon oil on its pure form, and 0.5, 1.0, and 5.0% of the nanostructured form. 10% cinnamon oil (pure form) showed 100% efficacy, whereas concentrations of 1 and 5% were 62 and 97% efficacious, respectively. Nanocapsules and nanoemulsions containing cinnamon oil at 5% showed 95 and 97% efficacy, respectively. In vivo (stage II)-16 naturally tick-infested cows were divided into four groups of four animals each: Group A was composed of dairy cows sprayed with Triton (control); Group B was composed of dairy cows sprayed with cinnamon oil in its pure form (5%), whereas groups C and D were composed of dairy cows sprayed with nanocapsules and nanoemulsions, respectively, containing cinnamon oil at 0.5%. The ticks on each animal were counted on days 0, 1, 4 and 20 after spraying. Animals sprayed with pure and nanoencapsulated cinnamon oil carried significantly fewer ticks on days 1 and 4 post-treatment and were free of ticks on day 20 post-treatment. Ticks collected from these dairy cows (24 h after application) had impaired oviposition and larval inhibition, resulting in 90.5 and 100% efficacy when using pure and nanocapsules, respectively. In conclusion, the pure and nanostructured forms of cinnamon oil interfered with tick reproduction, whereas a significant acaricidal effect was found when applied onto cattle.

  10. Association between entomopathogenic nematodes and fungi for control of Rhipicephalus microplus (Acari: Ixodidae).

    PubMed

    Monteiro, Caio Márcio Oliveira; Araújo, Laryssa Xavier; Matos, Renata Silva; da Silva Golo, Patrícia; Angelo, Isabele Costa; de Souza Perinotto, Wendell Marcelo; Coelho Rodrigues, Camila Aparecida; Furlong, John; Bittencourt, Vânia Rita Elias Pinheiro; Prata, Márcia Cristina Azevedo

    2013-10-01

    The aim of the study was to assess the effect of the association of entomopathogenic nematodes and fungi on Rhipicephalus microplus. The nematodes used were Heterorhabditis bacteriophora HP88 and Heterorhabditis indica LPP1 and the fungi were Metarhizium anisopliae IBCB 116 and Beauveria bassiana ESALQ 986. In the groups treated with the fungi, the females were immersed for 3 min in a conidial suspension, while in the groups treated with the nematodes, the ticks were exposed to infective juveniles. To evaluate the interaction between entomopathogens, the females were first immersed in a conidial suspension and then exposed to the nematodes. The egg mass weight and hatching percentage values of the groups treated with M. anisopliae IBCB 116 and B. bassiana ESALQ 986 in the two experiments were statistically similar (p > 0.05) to the values of the control group. In the groups treated only with nematodes, there was a significant reduction (p < 0.05) in the egg mass weight, a fact also observed for the hatching percentage of the group treated with H. indica LPP1. In all the groups treated with nematodes in association with fungi, there was a significant reduction (p < 0.05) in the egg mass weight and hatching percentage. The percentage of control of the groups treated with fungi alone varied from 31 to 55%. In the groups treated with nematodes associated or not with fungi, the control percentage was always greater than 90% and reached 100% in the group treated with H. bacteriophora HP88 associated with the fungus M. anisopliae IBCB 116.

  11. The current status of resistance to alpha-cypermethrin, ivermectin, and amitraz of the cattle tick (Rhipicephalus microplus) in Ecuador

    PubMed Central

    Garcés-Carrera, Sandra; Vanwambeke, Sophie O.; Madder, Maxime; Benítez-Ortiz, Washington

    2017-01-01

    Rhipicephalus microplus is widely distributed in tropical and subtropical areas of the world where livestock is a principal activity with great veterinary and economic importance. In Ecuador, this hematophagous ectoparasite has been observed between 0 and 2600 masl. One of the main tick control measures is the use of acaricides, which have been indiscriminately used worldwide and in Ecuador. In this country, no studies on acaricide resistance in Rhipicephalus microplus have been published. The current study aims to characterise the level of resistance of R. microplus against three main acaricides commonly used in Ecuador i.e. amitraz, alpha-cypermethrin and ivermectin to determine the level and pattern of dose-responses for R. microplus in 12 field populations (farms). The level of acaricide resistance was evaluated using three different bioassays: adult immersion test (AIT), larval package test (LPT) and larval immersion test (LIT), as recommended by the FAO. The predictive dose-responses were analysed by binomial logistics regression of the larval survival rate (resistance). In general, we found resistance of 67% for amitraz; 50% for alpha-cypermethrin and from 25 to 42% for ivermectin in the twelve field populations analysed. Resistance levels were studied in larval and adult bioassays, respectively, which were slightly modified for this study. For larval bioassays based on corrected mortality i.e. high (above 51%), medium (21–50%) and low (11–20%) resistance, percentages less than 10% were considered as susceptible. For the adult test, two resistance levels were used i.e. high (more than 76%) and medium (51 to 75%) resistance. Percentages lower than 50% were considered as susceptible. In this context, for larval bioassays, amitraz showed 21%, 38% and 8% for high, medium and low resistance, respectively. Alpha-cypermethrin presented 8%, 4 and 38% for high, medium and low resistance, respectively. Ivermectin presented 8%, 25% and 8% for high, medium and low

  12. The current status of resistance to alpha-cypermethrin, ivermectin, and amitraz of the cattle tick (Rhipicephalus microplus) in Ecuador.

    PubMed

    Rodríguez-Hidalgo, Richar; Pérez-Otáñez, Ximena; Garcés-Carrera, Sandra; Vanwambeke, Sophie O; Madder, Maxime; Benítez-Ortiz, Washington

    2017-01-01

    Rhipicephalus microplus is widely distributed in tropical and subtropical areas of the world where livestock is a principal activity with great veterinary and economic importance. In Ecuador, this hematophagous ectoparasite has been observed between 0 and 2600 masl. One of the main tick control measures is the use of acaricides, which have been indiscriminately used worldwide and in Ecuador. In this country, no studies on acaricide resistance in Rhipicephalus microplus have been published. The current study aims to characterise the level of resistance of R. microplus against three main acaricides commonly used in Ecuador i.e. amitraz, alpha-cypermethrin and ivermectin to determine the level and pattern of dose-responses for R. microplus in 12 field populations (farms). The level of acaricide resistance was evaluated using three different bioassays: adult immersion test (AIT), larval package test (LPT) and larval immersion test (LIT), as recommended by the FAO. The predictive dose-responses were analysed by binomial logistics regression of the larval survival rate (resistance). In general, we found resistance of 67% for amitraz; 50% for alpha-cypermethrin and from 25 to 42% for ivermectin in the twelve field populations analysed. Resistance levels were studied in larval and adult bioassays, respectively, which were slightly modified for this study. For larval bioassays based on corrected mortality i.e. high (above 51%), medium (21-50%) and low (11-20%) resistance, percentages less than 10% were considered as susceptible. For the adult test, two resistance levels were used i.e. high (more than 76%) and medium (51 to 75%) resistance. Percentages lower than 50% were considered as susceptible. In this context, for larval bioassays, amitraz showed 21%, 38% and 8% for high, medium and low resistance, respectively. Alpha-cypermethrin presented 8%, 4 and 38% for high, medium and low resistance, respectively. Ivermectin presented 8%, 25% and 8% for high, medium and low

  13. Effect of oil-based formulations of acaripathogenic fungi to control Rhipicephalus microplus ticks under laboratory conditions.

    PubMed

    Camargo, Mariana G; Golo, Patrícia S; Angelo, Isabele C; Perinotto, Wendell M S; Sá, Fillipe A; Quinelato, Simone; Bittencourt, Vânia R E P

    2012-08-13

    The formulations of acaripathogenic fungi to control ticks have been widely studied. The present study evaluated the efficacy of oil-based formulations of Metarhizium anisopliae sensu lato (s.l.), isolate Ma 959, and Beauveria bassiana, isolate Bb 986, on different Rhipicephalus microplus stages, comparing the efficacy between aqueous suspensions and 10, 15 and 20% mineral oil formulations. Twelve groups were formed: one aqueous control group; three mineral oil control groups, at 10, 15 or 20%; two aqueous fungal suspensions of M. anisopliae s.l. or B. bassiana; and three formulations of M. anisopliae (s.l.) or B. bassiana containing 10, 15, and 20% mineral oil. To prepare aqueous suspensions and oily formulations, fungal isolates were cultivated on rice grains in polypropylene bags. The conidial suspensions and formulations had a concentration of 10(8)conidia/mL. Bioassays were repeated twice. After treatment, the following biological parameters of engorged females were evaluated: hatching percentage, egg production index, nutritional index, and percentage of tick control. The following parameters were evaluated in the bioassays with eggs: period of incubation, period of hatch, and hatching percentage. Mortality was evaluated in bioassays with larvae. M. anisopliae s.l. and B. bassiana oil-based formulations were more effective than aqueous suspensions against R. microplus eggs, larvae and engorged females, however, there was no significant difference between the three oil concentrations used. M. anisopliae s.l. and B. bassiana formulated in mineral oil reached 93.69% and 21.67% efficacy, respectively, while M. anisopliae s.l. and B. bassiana aqueous suspensions attained 18.70% and 1.72% efficacy, respectively. M. anisopliae s.l. oil-based formulations caused significant effects in all biological parameters of engorged females while B. bassiana oil-based formulations modified significantly the nutritional index only. Eggs treated with M. anisopliae s.l. and B

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  16. Distribution of cattle fever tick infestations detected in the U.S. along the Texas/Mexico border

    USDA-ARS?s Scientific Manuscript database

    Species identification and coordinates of geographical premises for infestations of cattle fever ticks, Rhipicephalus (Boophilus) annulatus (Say) and R. (B.) microplus (Canestrini) were determined for 782 specimens submitted to the National Veterinary Services Laboratory during the eleven years betw...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  18. 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

  19. 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.

  20. 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.

  1. Acaricidal Activity of Petroleum Ether Extract of Leaves of Tetrastigma leucostaphylum (Dennst.) Alston against Rhipicephalus (Boophilus) annulatus

    PubMed Central

    Krishna, T. P. Adarsh; Krishna, T. P. Ajeesh; Chithra, N. D.; Deepa, P. E.; Darsana, U.; Sreelekha, K. P.; Juliet, Sanis; Nair, Suresh N.; Ajith Kumar, K. G.; 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. PMID:25436226

  2. 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.

  3. 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.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  6. Development and validation of a dual luciferase reporter system for in vitro evaluation of gene silencing efficacy in the Southern cattle tick: comparison to in vivo gene silencing by microinjection

    USDA-ARS?s Scientific Manuscript database

    The Southern cattle tick, Rhipicephalus (Boophilus) microplus, vectors bovine babesiosis and anaplasmosis, and was eradicated from the United States over several decades by the Cattle Fever Tick Eradication Program (CFTEP); however, R. microplus is endemic in Mexico and remains a continuing threat t...

  7. 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.

  8. 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.

  9. Immune and biochemical responses in skin differ between bovine hosts genetically susceptible and resistant to the cattle tick Rhipicephalus microplus.

    PubMed

    Franzin, Alessandra Mara; Maruyama, Sandra Regina; Garcia, Gustavo Rocha; Oliveira, Rosane Pereira; Ribeiro, José Marcos Chaves; Bishop, Richard; Maia, Antônio Augusto Mendes; Moré, Daniela Dantas; Ferreira, Beatriz Rossetti; Santos, Isabel Kinney Ferreira de Miranda

    2017-01-31

    Ticks attach to and penetrate their hosts' skin and inactivate multiple components of host responses in order to acquire a blood meal. Infestation loads with the cattle tick, Rhipicephalus microplus, are heritable: some breeds carry high loads of reproductively successful ticks, whereas in others, few ticks feed and reproduce efficiently. In order to elucidate the mechanisms that result in the different outcomes of infestations with cattle ticks, we examined global gene expression and inflammation induced by tick bites in skins from one resistant and one susceptible breed of cattle that underwent primary infestations with larvae and nymphs of R. microplus. We also examined the expression profiles of genes encoding secreted tick proteins that mediate parasitism in larvae and nymphs feeding on these breeds. Functional analyses of differentially expressed genes in the skin suggest that allergic contact-like dermatitis develops with ensuing production of IL-6, CXCL-8 and CCL-2 and is sustained by HMGB1, ISG15 and PKR, leading to expression of pro-inflammatory chemokines and cytokines that recruit granulocytes and T lymphocytes. Importantly, this response is delayed in susceptible hosts. Histopathological analyses of infested skins showed inflammatory reactions surrounding tick cement cones that enable attachment in both breeds, but in genetically tick-resistant bovines they destabilized the cone. The transcription data provided insights into tick-mediated activation of basophils, which have previously been shown to be a key to host resistance in model systems. Skin from tick-susceptible bovines expressed more transcripts encoding enzymes that detoxify tissues. Interestingly, these enzymes also produce volatile odoriferous compounds and, accordingly, skin rubbings from tick-susceptible bovines attracted significantly more tick larvae than rubbings from resistant hosts. Moreover, transcripts encoding secreted modulatory molecules by the tick were significantly more

  10. 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.

  11. 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.

  12. 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).

  13. 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.

  14. Evaluation of the Efficacy of Acaricides Used to Control the Cattle Tick, Rhipicephalus microplus, in Dairy Herds Raised in the Brazilian Southwestern Amazon

    PubMed Central

    Brito, Luciana G.; Barbieri, Fábio S.; Rocha, Rodrigo B.; Oliveira, Márcia C. S.; Ribeiro, Elisana Sales

    2011-01-01

    The adult immersion test (AIT) was used to evaluate the efficacy of acaricide molecules used for control of Rhipicephalus microplus on 106 populations collected in five municipalities in the state of Rondônia in the Brazilian South Occidental Amazon region. The analysis of the data showed that the acaricide formulations had different efficacies on the tick populations surveyed. The synthetic pyrethroids (SPs) acaricides were the least effective (48.35–76.84%), followed by SP + organophosphate (OP) associations (68.91–81.47%) and amidine (51.35–100%). For the macrocyclic lactones (MLs), the milbemycin (94.84–100%) was the most effective, followed by spinosad (93.21–100%) and the avermectins (81.34–100%). The phenylpyrazole (PZ) group had similar efficacy (99.90%) to the MLs. Therefore, SP acaricides, including associations with OP, and formulations based on amidine presented low in vitro efficacy to control the R. microplus populations surveyed. PMID:21547224

  15. Heterorhabditis bacteriophora (Rhabditida: Heterorhabditidae) HP88 for biological control of Rhipicephalus microplus (Acari: Ixodidae): the effect of different exposure times of engorged females to the nematodes.

    PubMed

    Monteiro, Caio Márcio de Oliveira; Prata, Márcia Cristina de Azevedo; Faza, Aline; Batista, Elder Simões de Paula; Dolinski, Cláudia; Furlong, John

    2012-04-30

    The aim of this study was to assess the influence of different exposure times of engorged female the Rhipicephalus microplus to infective juveniles of Heterorhabditis bacteriophora isolate HP88. The engorged females were divided into seven groups (six treatments and one control) of 20 ticks each with statistically similar average weights (p>0.05) and exposed to concentrations of 300 nematodes/tick for periods of 3, 6, 12, 24, 48 and 72 h. The following biological parameters were monitored: pre-oviposition period; egg mass weight; hatching percentage; and efficacy of treatment. There was no influence of the exposure time on the pre-oviposition period (p>0.05), while there were significant differences (p<0.05) between the groups exposed for 24, 48 and 72 h and the control group with respect to the egg mass weight, and 24h for hatching percentage. Treatment efficacy reached 100% after exposure for 48 and 72 h. These results demonstrate that infective juveniles of H. bacteriophora HP88, under laboratory conditions, interfere in the majority of the biological parameters of the non-parasitic phase of engorged R. microplus females when the exposure time is greater than or equal to 24h. Copyright © 2011 Elsevier B.V. All rights reserved.

  16. 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.

  17. 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).

  18. Immune recognition of salivary proteins from the cattle tick Rhipicephalus microplus differs according to the genotype of the bovine host.

    PubMed

    Garcia, Gustavo Rocha; Maruyama, Sandra Regina; Nelson, Kristina T; Ribeiro, José Marcos Chaves; Gardinassi, Luiz Gustavo; Maia, Antonio Augusto Mendes; Ferreira, Beatriz Rossetti; Kooyman, Frans N J; de Miranda Santos, Isabel K F

    2017-03-14

    Males of the cattle tick Rhipicephalus microplus produce salivary immunoglobulin-binding proteins and allotypic variations in IgG are associated with tick loads in bovines. These findings indicate that antibody responses may be essential to control tick infestations. Infestation loads with cattle ticks are heritable: some breeds carry high loads of reproductively successful ticks, in others, few ticks feed and they reproduce inefficiently. Different patterns of humoral immunity against tick salivary proteins may explain these phenotypes. We describe the profiles of humoral responses against tick salivary proteins elicited during repeated artificial infestations of bovines of a tick-resistant (Nelore) and a tick-susceptible (Holstein) breed. We measured serum levels of total IgG1, IgG2 and IgE immunoglobulins and of IgG1 and IgG2 antibodies specific for tick salivary proteins. With liquid chromatography followed by mass spectrometry we identified tick salivary proteins that were differentially recognized by serum antibodies from tick-resistant and tick-susceptible bovines in immunoblots of tick salivary proteins separated by two-dimensional electrophoresis. Baseline levels of total IgG1 and IgG2 were significantly higher in tick-susceptible Holsteins compared with resistant Nelores. Significant increases in levels of total IgG1, but not of IgG2 accompanied successive infestations in both breeds. Resistant Nelores presented with significantly higher levels of salivary-specific antibodies before and at the first challenge with tick larvae; however, by the third challenge, tick-susceptible Holsteins presented with significantly higher levels of IgG1 and IgG2 tick salivary protein-specific antibodies. Importantly, sera from tick-resistant Nelores reacted with 39 tick salivary proteins in immunoblots of salivary proteins separated in two dimensions by electrophoresis versus only 21 spots reacting with sera from tick-susceptible Holsteins. Levels of tick saliva

  19. A novel amino acid substitution in the para-sodium channel gene in Rhipicephalus microplus (Acari: Ixodidae) associated with knockdown resistance.

    PubMed

    Aguirre, Marcelino; Flores, Adriana E; Alvarez, Genoveva; Molina, Alberto; Rodriguez, Iram; Ponce, Gustavo

    2010-12-01

    Resistance acquired by the tick Rhipicephalus microplus (Canestrini) to different types of ixodicides in Mexico has had a negative impact on national and local livestock, mainly due to the transmission of diseases such as babesiosis and anaplasmosis, among others. The technique used for the diagnosis of resistance was that in the bioassays noted in the Norma Oficial Mexicana (NOM-006-ZOO-1994). The purpose of this investigation was the determination of resistance to pyrethroids through isoleucine-phenylalanine mutation in the gene KDR, in a population of ticks from Montemorelos, NL, Mexico. Preliminary bioassays demonstrated resistance to cypermethrin and deltamethrin (27.4%) and flumethrin (36.7-34.7%). To identify the mutation, DNA was extracted from 100 mg of larvae (pools), 10 pools were assessed by PCR, in which a pair of primers designed with the program Oligo 2.0 and Amplify 1.2 amplified a 136 bp fragment containing the mutation. The PCR product was subsequently sequenced to confirm the presence of the mutation. A strain susceptible to pyrethroid insecticides (Mora strain) was used as control, but it did not show the mutation. However, the mutation was detected in 4 out of 10 samples of the strain Montemorelos.

  20. Lab-on-a-chip and SDS-PAGE analysis of hemolymph protein profile from Rhipicephalus microplus (Acari: Ixodidae) infected with entomopathogenic nematode and fungus.

    PubMed

    Golo, Patrícia Silva; Dos Santos, Alessa Siqueira de Oliveira; Monteiro, Caio Marcio Oliveira; Perinotto, Wendell Marcelo de Souza; Quinelato, Simone; Camargo, Mariana Guedes; de Sá, Fillipe Araujo; Angelo, Isabele da Costa; Martins, Marta Fonseca; Prata, Marcia Cristina de Azevedo; Bittencourt, Vânia Rita Elias Pinheiro

    2016-09-01

    In the present study, lab-on-a-chip electrophoresis (LoaC) was suggested as an alternative method to the conventional polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE) to analyze raw cell-free tick hemolymph. Rhipicephalus microplus females were exposed to the entomopathogenic fungus Metarhizium anisopliae senso latu IBCB 116 strain and/or to the entomopathogenic nematode Heterorhabditis indica LPP1 strain. Hemolymph from not exposed or exposed ticks was collected 16 and 24 h after exposure and analyze by SDS-PAGE or LoaC. SDS-PAGE yielded 15 bands and LoaC electrophoresis 17 bands. Despite the differences in the number of bands, when the hemolymph protein profiles of exposed or unexposed ticks were compared in the same method, no suppressing or additional bands were detected among the treatments regardless the method (i.e., SDS-PAGE or chip electrophoresis using the Protein 230 Kit®). The potential of LoaC electrophoresis to detect protein bands from tick hemolymph was considered more efficient in comparison to the detection obtained using the traditional SDS-PAGE method, especially when it comes to protein subunits heavier than 100 KDa. LoaC electrophoresis provided a very good reproducibility, and is much faster than the conventional SDS-PAGE method, which requires several hours for one analysis. Despite both methods can be used to analyze tick hemolymph composition, LoaC was considered more suitable for cell-free hemolymph protein separation and detection. LoaC hemolymph band percent data reported changes in key proteins (i.e., HeLp and vitellogenin) exceptionally important for tick embryogenesis. This study reported, for the first time, tick hemolymph protein profile using LoaC.

  1. 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.

  2. 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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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”, ...

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

    USDA-ARS?s Scientific Manuscript database

    The southern cattle tick (Rhipicephalus (Boophilus) microplus) has historically been a devastating pest to the cattle industry worldwide. The use of chemical acaricides has been the mainstay for controlling the southern cattle tick. However, there have been several reports of chemical acaricide resi...

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

    USDA-ARS?s Scientific Manuscript database

    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 popul...

  10. Comparative daily activity patterns of Nilgai, Boselaphus tragocamelus and white-tailed deer, Odocoileus virginianus in South Texas

    USDA-ARS?s Scientific Manuscript database

    Nilgai antelope, Boselaphus tragocamelus, and white-tailed deer (WTD), Odocoileus virginianus, are hosts of the cattle fever ticks, Rhipicephalus (Boophilus) microplus, and R. (B.) annulatus in south Texas. Daily activity patterns were studied to develop optimum timing for field treatment methods fo...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  12. Facilitative ecological interactions between invasive species: Arundo donax (Poaceae) stands as favorable habitat for cattle ticks (Acari: Ixodidae) along the U.S.-Mexico border

    USDA-ARS?s Scientific Manuscript database

    The southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini), is a key vector of protozoa that causes bovine babesiosis. Largely eradicated from most of the U.S., the cattle tick continues to infest the Cattle Fever Tick Quarantine Zone in south Texas. Management areas of the souther...

  13. Proportion of White-tailed deer using medicated bait sites in Southern Texas

    USDA-ARS?s Scientific Manuscript database

    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 Eradication Program. Our objective was to assess patterns of deer visitation to medicated bait...

  14. White-tailed Deer Visitation Rates at Medicated Bait Sites in Southern Texas

    USDA-ARS?s Scientific Manuscript database

    The cattle fever tick, Rhipicephalus (Boophilus) microplus, has been found on white-tailed deer (Odocoileus virginianus) complicating eradication efforts of the USDA’s Cattle Fever Tick Eradication Program. Our objective was to assess patterns of deer visitation to medicated bait sites used to treat...

  15. Genetic basis and impact of tick acaricide resistance

    USDA-ARS?s Scientific Manuscript database

    Acaricide resistance in the cattle tick, Boophilus microplus, has been studied for the last 20 years from the toxicology, metabolic, and genomic points of view, however, only few methods for molecular detection of resistance have been developed. Despite the relatively poor sensitivity for resistance...

  16. Invasive plant-invasive insect interactions: Giant reed invasions as suitable refuge for cattle fever ticks (Acari: Ixodidae)

    USDA-ARS?s Scientific Manuscript database

    Though largely eradicated from the U.S. for the past half century, the reemergence of populations of southern cattle tick (Rhipicephalus (Boophilus) microplus) is of major concern to the U.S. cattle industry. Southern cattle ticks are vectors of blood parasites that cause a lethal cattle disease, es...

  17. 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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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 (...

  19. 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. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. 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.

  1. Community structure, seasonal variations and interactions between native and invasive cattle tick species in Benin and Burkina Faso.

    PubMed

    Biguezoton, Abel; Adehan, Safiou; Adakal, Hassane; Zoungrana, Sébastien; Farougou, Souaïbou; Chevillon, Christine

    2016-01-27

    The variation of tick abundance on ruminants had received little attention in West Africa before Rhipicephalus (Boophilus) microplus started to invade this region in the early 2000s. Ten years later, R. microplus was suspected to have replaced the native ticks. In addition to testing this hypothesis, this study investigated the interactions between native and invasive ticks and the relative role of climatic and geographical variables in the variations of tick community composition (beta diversity) on cattle herds. A one-year-long survey was performed in Benin and Burkina Faso during which adult ticks were collected from 144 steers from 12 localities in four different areas once a month. Morphological features were used to assign the collected ticks to different species (A. variegatum, R. annulatus, R. decoloratus, R. microplus and R. geigyi). Beta diversity analyses and generalized linear models allowed characterizing the geographical variations in species assemblage and the effect of co-infestation patterns on the seasonal variations in the abundance and incidence rates of each taxon. About 68% (22,491/32,148) of all the adult ticks collected in one year were R. microplus. The most heterogeneously distributed taxa were Hyalomma spp and R. microplus and the lowest specific diversity was found in Central Burkina Faso. Although climatic variables did not provide any additional information on the variation in species assemblages compared with the sampling geography, adult tick abundance tended to peak during the late (Boophilus subgenus) or early (other taxa) rainy season. In most taxon-per-locality analyses, the abundance and incidence rate of a given tick taxon significantly increased when the host was co-infested by other taxa. The comparison with previous estimates (when possible) did not support the hypothesis that R. microplus invasion led to a decrease in native tick species abundance. The co-infestation patterns among native and invasive tick species are key

  2. Seasonal variation of ticks (Ixodidae) in Bos taurus x Bos indicus cattle under rotational grazing in forested and deforested habitats in northwestern Argentina.

    PubMed

    Mangold, A J; Aguirre, D H; Gaido, A B; Guglielmone, A A

    1994-09-01

    The seasonal occurrence of ticks infesting cattle was monitored in a Bos taurus x Bos indicus commercial herd fed on a combination of native grasses and annual pastures in northwestern Argentina. The number of female ticks (Boophilus and Amblyomma) and the proportion of cattle infested with larvae and nymphs of Amblyomma ticks were recorded from a sample of 15 cows in 24 visits at intervals of 21-50 days from 5 December 1986 to 24 November 1988. The cattle were maintained on native grasses in forested habitats for 341 days, on native grasses in deforested habitats for 116 days and finally for 264 days on annual pastures. On 15 visits Boophilus microplus females were found, Amblyomma cajennense were detected on 11 counts, seven counts were positive for Amblyomma neumanni and two counts for Amblyomma parvum females. The peak of abundance of Boophilus microplus was found to be at the end of March 1987 with a mean of 36 females per side of cow. The peak numbers of A. cajennense (20 ticks), A. neumanni (12 ticks) and A. parvum (nine ticks) were found in December 1987, July 1987 and November 1988, respectively. The peak of Boophilus microplus was detected while cattle were on natural grasses in deforested habitats, peaks of other tick species were found when cattle were on natural grasses in forested habitats. The maximum cattle infestations with Amblyomma larvae (34% of cattle, May 1987) and nymphs (100%, August 1987) were detected while cattle were on natural grasses in forested habitats.(ABSTRACT TRUNCATED AT 250 WORDS)

  3. Proteomics Approach to the Study of Cattle Tick Adaptation to White Tailed Deer

    PubMed Central

    Popara, Marina; Villar, Margarita; Mateos-Hernández, Lourdes; Fernández de Mera, Isabel G.; de la Fuente, José

    2013-01-01

    Cattle ticks, Rhipicephalus (Boophilus) microplus, are a serious threat to animal health and production. Some ticks feed on a single host species while others such as R. microplus infest multiple hosts. White tailed deer (WTD) play a role in the maintenance and expansion of cattle tick populations. However, cattle ticks fed on WTD show lower weight and reproductive performance when compared to ticks fed on cattle, suggesting the existence of host factors that affect tick feeding and reproduction. To elucidate these factors, a proteomics approach was used to characterize tick and host proteins in R. microplus ticks fed on cattle and WTD. The results showed that R. microplus ticks fed on cattle have overrepresented tick proteins involved in blood digestion and reproduction when compared to ticks fed on WTD, while host proteins were differentially represented in ticks fed on cattle or WTD. Although a direct connection cannot be made between differentially represented tick and host proteins, these results suggested that differentially represented host proteins together with other host factors could be associated with higher R. microplus tick feeding and reproduction observed in ticks fed on cattle. PMID:24364032

  4. 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.

  5. Tick Cell Lines for Study of Crimean-Congo Hemorrhagic Fever Virus and Other Arboviruses

    PubMed Central

    Kohl, Alain; Bente, Dennis A.; Fazakerley, John K.

    2012-01-01

    Abstract Continuous cell lines derived from many of the vectors of tick-borne arboviruses of medical and veterinary importance are now available. Their role as tools in arbovirus research to date is reviewed and their potential application in studies of tick cell responses to virus infection is explored, by comparison with recent progress in understanding mosquito immunity to arbovirus infection. A preliminary study of propagation of the human pathogen Crimean-Congo hemorrhagic fever virus (CCHFV) in tick cell lines is reported; CCHFV replicated in seven cell lines derived from the ticks Hyalomma anatolicum (a known vector), Amblyomma variegatum, Rhipicephalus (Boophilus) decoloratus, Rhipicephalus (Boophilus) microplus, and Ixodes ricinus, but not in three cell lines derived from Rhipicephalus appendiculatus and Ornithodoros moubata. This indicates that tick cell lines can be used to study growth of CCHFV in arthropod cells and that there may be species-specific restriction in permissive CCHFV infection at the cellular level. PMID:21955214

  6. 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.

  7. Ehrlichia ruminantium grows in cell lines from four ixodid tick genera.

    PubMed

    Bell-Sakyi, L

    2004-05-01

    Continuous cell lines from the ticks Amblyomma variegatum, Boophilus decoloratus, Boophilus microplus, Hyalomma anatolicum anatolicum, Ixodes scapularis, Ixodes ricinus and Rhipicephalus appendiculatus were tested for ability to support growth of the rickettsial pathogen Ehrlichia (previously Cowdria) ruminantium. Five E.ruminantium isolates, from West Africa, South Africa and the French West Indies, were used. Twelve tick cell lines were inoculated with E.ruminantium derived either from cultures of a bovine endothelial cell strain designated BPC or from other tick cell lines. Successful infection resulted in either continuous growth (in which the pathogen/cell line system could be perpetuated through regular subculture on fresh, uninfected cells for many months or years) or finite growth (in which the pathogen disappeared after one or a few subcultures). Infection with E.ruminantium from BPC was established in I.scapularis, I.ricinus and A.variegatum cell lines; E.ruminantium was transferred from these infected cell lines to B.decoloratus, B.microplus and R. appendiculatus cell lines. H.a.anatolicum cells could not be infected with E.ruminantium by any procedure. All five E.ruminantium isolates grew continuously in at least one tick cell line at temperatures between 28 degrees C and 37 degrees C; three of the isolates were successfully re-established in BPC following prolonged maintenance in tick cells. This study demonstrates that E.ruminantium is not intrinsically restricted to growth in cells from ticks of the natural vector genus Amblyomma.

  8. Advances in tick vaccinology in Brazil: from gene expression to immunoprotection.

    PubMed

    Andreotti, Renato; Giachetto, Poliana Fernanda; Cunha, Rodrigo Casquero

    2018-01-01

    Rhipicephalus (Boophilus) microplus has substantial economic impact on the cattle breeding industry and, chemical control and tick resistance development are the major concern. There is a worldwide search for new options, and control using vaccines has been the main focus nowadays. Studies performed in Brazil found that Bm86-based immunization of bovines reduced the infestation of R. (B.) microplus of vaccinated bovines by 45% to 60%. Native Boophilusmicroplus tripsin inhibitors (BmTIs) with trypsin-, kallikrein-, and elastase-inhibiting activities have been used as immunogens in bovines reaching 72.8.% of efficacy. The reverse vaccinology approach has also been used for antigen search using transcriptome analysis to identify and characterize potential antigens. Study has generated more than 600 million sequences using RNA-seq of larvae, nymphs, salivary glands, intestines, and ovaries of the tick R. (B) microplus. Based on the set of transcripts obtained using this strategy, a total of 20,326 protein sequences have been identified. A pipeline analysis built in house identified the protein sequences that were most likely to be immunogenic based on the overall structural characteristic analysis.

  9. 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.

  10. In vitro acaricide activity of Ocotea aciphylla (Nees) Mez. (Lauraceae) extracts and identification of the compounds from the active fractions.

    PubMed

    Souza Conceição, Rodrigo; de A Carneiro, Monique Marylin A; Alves Reis, Isabella Mary; Branco, Alexsandro; Curcino Vieira, Ivo Jose; Braz-Filho, Raimundo; Borges Botura, Mariana

    2017-02-01

    The in vitro acaricide activity of hexane, ethyl acetate and ethanol extracts of Ocotea aciphylla leaves was investigated by means of an immersion tests using Rhipicephalus (Boophilus) microplus engorged females and larvae. All extracts were shown effective against the different stages of the parasite, and the ethanol extract (50mg/mL concentration) was the most active (more than 90% efficacy in both assays). The ethanolic extract was fractionated using column chromatography with silica gel as stationary phase to furnish several fractions that were tested against larvae of R. (B.) microplus. Three fractions showed high acaricidal activity (50mg/mL), causing between 84.2% and 100% mortality of the larvae. The anticholinesterase effect of these fractions was determined spectrophotometrically using a microtiter assay. The chemical investigation of the active fractions led to the characterization of terpenoids (cadalene 1 and squalene 2), a phenylpropanoid (dillapiole 3) and a phenolic mixture containing five proanthocyanidins (4-8) and a flavonoid(vitexin-2"-O-rhamnoside 9). Our findings suggest that the O. aciphylla has potent acaricidal activity in vitro against R. (B.) microplus, and that different secondary metabolites are responsible for this effect. Copyright © 2016 Elsevier GmbH. All rights reserved.

  11. 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.

  12. Ticks on livestock in St. Lucia.

    PubMed

    Garris, G I; Scotland, K

    1985-12-01

    Cattle, sheep, goats and horses were examined for ticks. Over 95% of Holstein cross-breeds, 28% of sheep (local mixed breeds) and 18% of goats (local mixed breeds) examined from 18 August to 4 September 1983 were infested with the southern cattle tick, Boophilus microplus Canestrini. About 90 and 17% of the horses examined were infested with the tropical horse tick, Anocentor nitens Neumann, and the tropical bont tick, Amblyomma variegatum Fabricius, respectively. The tropical bont tick was found infesting 10% of cattle in the Gros Islet area of St. Lucia. The tropical bont tick was also found associated with a severe skin disease, dermatophilosis, caused by the bacterium Dermatophilus congolensis, in 54% of the cattle infested by A. variegatum in the Gros Islet and Dauphin areas of St. Lucia.

  13. Low level deltamethrin resistance in ticks from cattle of Kerala, a south Indian state.

    PubMed

    Jyothimol, G; Ravindran, R; Juliet, S; Ajithkumar, K G; Suresh, N N; Vimalkumar, M B; Lenka, D R; Varghese, S; Ghosh, Srikanta

    2014-08-29

    The deltamethrin resistance status in Rhipicephalus (Boophilus) annulatus and R. (B.) microplus ticks collected from cattle of five organized farms of Kerala, south India was evaluated. Resistance was characterized using biological (larval packet test), biochemical (esterase enzyme activity assay) and molecular tools (PCR amplification and sequencing of deltamethrin resistance-associated genes). Characterization of field isolates revealed level I resistance in ticks collected from four out of five farms. Elevated level of α/β esterase activity was not recorded in isolates showing level I resistance. Previously reported point mutations in the carboxyl esterase (G1120A) and sodium channel (T2134A and C190A) genes were not observed in any of the field isolates. The present study showed a low level (level I) resistance is developed in the most economically important ticks infesting cattle of this state and it cautions the development of large scale resistance in future. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Babesiasis in Nanjing area, China.

    PubMed

    Shen, Y; Gao, J; Xu, K; Xue, L; Zhang, Y; Shi, B; Li, D; Wei, X; Higuchi, S

    1997-11-01

    This article discusses the tick-borne babesiases which harm dairy cattle, buffaloes and dogs. In addition, the pathogen, vector, seasonal occurrence, hosts, symptoms, pathological changes and treatment are summarized and analysed. The pathogens in this area were identified as Babesia bigemina, B. bovis and B. gibsoni; the vectors as Boophilus microplus, Rhipicephalus haemaphysaloides, Haemaphysalis longicornis. Affected buffaloes were found from April to September, peak numbers occurred between May and July. Affected dairy cattle were most numerous in July and August. Hunting dogs were the most affected dogs. Infected dogs were found from February until the beginning of December; peak numbers occurred from September to November. Affected animals were acute cases. Symptoms and pathological changes were obvious, but jaundice was rare in dogs. Acaprin, acriflavine, imidocarb and berenil were used early and late. Their effect was ideal but resistance has developed to these drugs.

  15. 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.

  16. Failure of vaccine strains of Babesia bovis to regain infectivity for ticks during long-standing infections in cattle.

    PubMed

    Dalgliesh, R J; Stewart, N P

    1977-09-01

    Two strains of Babesia bovis that were known to have lost infectivity for the normal tick vector, Boophilus microplus, due to repeated blood passaging in cattle, were studied to determine whether the strains would regain infectivity for ticks during longstanding infections. Parasitaemias were monitored in 4 chronically infected calves that were regularly infested with ticks. Two strains of ticks known to be susceptible to infection with unmodified strains of B. bovis were used. Adult female ticks that dropped from the calves on days that a parasitaemia was evident were tested for B. bovis infection. Sixty-six batches of ticks collected up to 279 days after infection of the calves produced 14 pools of larvae, none of which transmitted infection. Primary infections established from the chronic infections by subinoculation at 200, 259 and 333 days after infection of the calves were also not transmitted by ticks.

  17. Expression of intracellular calcium signalling genes in cattle skin during tick infestation.

    PubMed

    Bagnall, N; Gough, J; Cadogan, L; Burns, B; Kongsuwan, K

    2009-04-01

    It is widely acknowledged that changes in intracellular calcium ion (Ca(2+)) concentration provide dynamic signals that control a plethora of cellular processes, including triggering and mediating host defence mechanisms. In this study, quantitative real-time PCR was used to analyse gene expression of 14 Ca(2+) signalling proteins in skin obtained from high tick-resistant (HR) and low tick-resistant (LR) cattle following artificial challenge with cattle tick (Rhipicephalus (Boophilus) microplus). Up-regulation of numerous genes was observed in both HR and LR skin following tick challenge, however substantially higher transcription activation was found in HR tissue. The elevated expression in HR skin of specific Ca(2+) signalling genes such as AHNAK, CASQ, IL2, NFAT2CIP and PLCG1 may be related to host resistance. Our data suggest that Ca(2+) and its associated proteins might play an important role in host response to ticks and that further investigation is warranted.

  18. 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.

  19. Bm86 homologues and novel ATAQ proteins with multiple epidermal growth factor (EGF)-like domains from hard and soft ticks☆

    PubMed Central

    Nijhof, Ard M.; Balk, Jesper A.; Postigo, Milagros; Rhebergen, Anne Marie; Taoufik, Amar; Jongejan, Frans

    2010-01-01

    Tick control on livestock relies principally on the use of acaricides but the development of acaricide resistance and concerns for environmental pollution underscore the need for alternative control methods, for instance through the use of anti-tick vaccines. Two commercial vaccines based on the recombinant Bm86 protein from Rhipicephalus (Boophilus) microplus ticks were developed. Partial protection of the Bm86 vaccine against other Rhipicephalus (Boophilus) and Hyalomma tick species suggests that the efficacy of a Bm86-based vaccine may be enhanced when based on the orthologous recombinant Bm86 antigen. We therefore identified and analysed the Bm86 homologues from species representing the main argasid and ixodid tick genera, including two from the prostriate Ixodes ricinus tick species. A novel protein from metastriate ticks with multiple epidermal growth factor (EGF)-like domains which is structurally related to Bm86 was identified by using a 3′ rapid amplification of cDNA ends (3′-RACE) method with a degenerate primer based on a highly conserved region of Bm86 and its orthologues. This second protein was named ATAQ after a part of its signature peptide. Quantitative reverse transcriptase-PCR showed that ATAQ proteins are expressed in both midguts and Malpighian tubules, in contrast to Bm86 orthologues which are expressed exclusively in tick midguts. Furthermore, expression of this protein over the life stages of R. microplus and Rhipicephalus appendiculatus was more continuous compared with Bm86. Although a highly effective vaccine antigen, gene silencing of Bm86 by RNA interference (RNAi) produced only a weak phenotype. Similarly the RNAi phenotype of Rhipicephalus evertsi evertsi females in which the expression of Ree86, ReeATAQ or a combination of both genes was silenced by RNAi did not differ from a mock-injected control group. The vaccine potential of ATAQ proteins against tick infestations is yet to be evaluated. PMID:20647015

  20. Wildlife as hosts for ticks (Acari) in Antigua, West Indies.

    PubMed

    Corn, J L; Kavanaugh, D M; Creekmore, T E; Robinson, J L

    1994-01-01

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

  1. 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.

  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. 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.

  4. Acaricidal, pediculicidal and larvicidal activity of synthesized ZnO nanoparticles using Momordica charantia leaf extract against blood feeding parasites.

    PubMed

    Gandhi, P Rajiv; Jayaseelan, C; Mary, R Regina; Mathivanan, D; Suseem, S R

    2017-10-01

    The aim of the present study was to evaluate the acaricidal, pediculicidal and larvicidal effect of synthesized zinc oxide nanoparticles (ZnO NPs) using Momordica charantia leaf extract against the larvae of Rhipicephalus (Boophilus) microplus, adult of Pediculus humanus capitis, and the larvae of Anopheles stephensi, Culex quinquefasciatus. The ZnO NPs were characterized by using UV, XRD, FTIR and SEM-EDX. The SEM image confirms that the synthesized nanoparticles were spherical in shape with a size of 21.32 nm. The results of GC-MS analysis indicates the presence of the major compound of Nonacosane (C29H60) in the M. charantia leaf extract. Cattle tick, head lice and mosquito larvae were exposed to a varying concentrations of the synthesized ZnO NPs and M. charantia leaf extract for 24 h. Compared to the leaf aqueous extract, biosynthesized ZnO NPs showed higher toxicity against R. microplus, P. humanus capitis, An. stephensi, and Cx. Quinquefasciatus with the LC50 values of 6.87, 14.38, 5.42, and 4.87 mg/L, respectively. The findings revealed that synthesized ZnO NPs possess excellent anti-parasitic activity. These results suggest that the green synthesized ZnO NPs has the potential to be used as an ideal ecofriendly approach for the control of R. microplus, P. humanus capitis and the mosquito larvae of An. Stephensi and Cx. quinquefasciatus. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Subolesin: a candidate vaccine antigen for the control of cattle tick infestations in Indian situation.

    PubMed

    Shakya, Mukesh; Kumar, Binod; Nagar, Gaurav; de la Fuente, José; Ghosh, Srikanta

    2014-06-12

    Identification of cross-protective tick vaccine antigens is a challenging area of veterinary research. To address this challenge, a recently identified candidate tick protective antigen, Subolesin (SUB), was targeted in this research. The conservation of subolesin ortholog of Hyalomma anatolicum and Rhipicephalus (Boophilus) microplus across different Indian strains was 98.1-99.4% (within species), while at the amino acid level SUB sequence homology was ≥53.2% (between tick species). Recombinant R. (B.) microplus SUB (rBmSu) was produced in Escherichia coli and characterized. Cross-bred cattle male calves (N=10) were immunized with three doses of 100 μg each of the rBmSu emulsified in 10% Montanide 888 at monthly intervals on days 0, 30 and 60. The control group was injected with PBS in 10% Montanide 888. For the first tick challenge, calves were infested with larvae of R. (B.) microplus generated from 100mg eggs 2 weeks after last immunization (day 75). The immunization resulted in 16.3%, 8.0%, 9.4%, and 26.1% reduction in female tick numbers (DT), weight (DW), oviposition (DO) and egg fertility (DF), respectively, when compared to controls. In the subsequent challenge on day 105, DT, DW, DO and DF were reduced by 9.0%, 4.1%, 8.6%, and 24.2%, respectively, when compared to controls. The vaccine efficacy (E) was equal to 44.0% and 37.2% after the first and second challenges, respectively. The results showed a positive correlation between antibody titers for both total IgG and IgG1 and E in the second but not in the first tick challenge. These results suggested the possibility of developing a SUB-based vaccine for control of cattle tick infestations under Indian conditions. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Invasive potential of cattle fever ticks in the southern United States.

    PubMed

    Giles, John R; Peterson, A Townsend; Busch, Joseph D; Olafson, Pia U; Scoles, Glen A; Davey, Ronald B; Pound, J Mathews; Kammlah, Diane M; Lohmeyer, Kimberly H; Wagner, David M

    2014-04-17

    For >100 years cattle production in the southern United States has been threatened by cattle fever. It is caused by an invasive parasite-vector complex that includes the protozoan hemoparasites Babesia bovis and B. bigemina, which are transmitted among domestic cattle via Rhipicephalus tick vectors of the subgenus Boophilus. In 1906 an eradication effort was started and by 1943 Boophilus ticks had been confined to a narrow tick eradication quarantine area (TEQA) along the Texas-Mexico border. However, a dramatic increase in tick infestations in areas outside the TEQA over the last decade suggests these tick vectors may be poised to re-invade the southern United States. We investigated historical and potential future distributions of climatic habitats of cattle fever ticks to assess the potential for a range expansion. We built robust spatial predictions of habitat suitability for the vector species Rhipicephalus (Boophilus) microplus and R. (B.) annulatus across the southern United States for three time periods: 1906, present day (2012), and 2050. We used analysis of molecular variance (AMOVA) to identify persistent tick occurrences and analysis of bias in the climate proximate to these occurrences to identify key environmental parameters associated with the ecology of both species. We then used ecological niche modeling algorithms GARP and Maxent to construct models that related known occurrences of ticks in the TEQA during 2001-2011 with geospatial data layers that summarized important climate parameters at all three time periods. We identified persistent tick infestations and specific climate parameters that appear to be drivers of ecological niches of the two tick species. Spatial models projected onto climate data representative of climate in 1906 reproduced historical pre-eradication tick distributions. Present-day predictions, although constrained to areas near the TEQA, extrapolated well onto climate projections for 2050. Our models indicate the potential

  7. Invasive potential of cattle fever ticks in the southern United States

    PubMed Central

    2014-01-01

    Abstract' Background For >100 years cattle production in the southern United States has been threatened by cattle fever. It is caused by an invasive parasite-vector complex that includes the protozoan hemoparasites Babesia bovis and B. bigemina, which are transmitted among domestic cattle via Rhipicephalus tick vectors of the subgenus Boophilus. In 1906 an eradication effort was started and by 1943 Boophilus ticks had been confined to a narrow tick eradication quarantine area (TEQA) along the Texas-Mexico border. However, a dramatic increase in tick infestations in areas outside the TEQA over the last decade suggests these tick vectors may be poised to re-invade the southern United States. We investigated historical and potential future distributions of climatic habitats of cattle fever ticks to assess the potential for a range expansion. Methods We built robust spatial predictions of habitat suitability for the vector species Rhipicephalus (Boophilus) microplus and R. (B.) annulatus across the southern United States for three time periods: 1906, present day (2012), and 2050. We used analysis of molecular variance (AMOVA) to identify persistent tick occurrences and analysis of bias in the climate proximate to these occurrences to identify key environmental parameters associated with the ecology of both species. We then used ecological niche modeling algorithms GARP and Maxent to construct models that related known occurrences of ticks in the TEQA during 2001–2011 with geospatial data layers that summarized important climate parameters at all three time periods. Results We identified persistent tick infestations and specific climate parameters that appear to be drivers of ecological niches of the two tick species. Spatial models projected onto climate data representative of climate in 1906 reproduced historical pre-eradication tick distributions. Present-day predictions, although constrained to areas near the TEQA, extrapolated well onto climate projections for

  8. 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

  9. 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.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

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

    USDA-ARS?s Scientific Manuscript database

    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...

  13. Molecular epidemiological surveillance to assess emergence and re-emergence of tick-borne infections in tick samples from China evaluated by nested PCRs.

    PubMed

    Yu, Pei-Fa; Niu, Qing-Li; Liu, Zhi-Jie; Yang, Ji-Fei; Chen, Ze; Guan, Gui-Quan; Liu, Guang-Yuan; Luo, Jian-Xun; Yin, Hong

    2016-06-01

    An investigation was performed to detect eight pathogens in ticks collected from grass tips or animals in the southern, central and northeast regions of China. DNA samples extracted from ticks were collected from ten different locations in eight provinces of China and subjected to screening for tick-borne pathogens, including Borrelia burgdorferi sensu lato, Ehrlichia spp., Rickettsia spp., Babesia/Theileria spp., Ehrlichia ruminantium, Coxiella burnetii, and Francisella tularensis, using nested PCR assays and sequencing analysis. The results indicated that Borrelia spp., Rickettsia spp., and Babesia/Theileria spp. were detected in all of the investigated provinces. Ehrlichia spp. was also found in all of the surveyed areas, except Guangxi, Luobei and Tonghe counties in Heilongjiang province. The average prevalence of these pathogens was 18.4% (95% CI=12.8-42.5), 60.3% (95% CI=18.2-65.3), 26.0% (95% CI=25.8-65.1), and 28.7% (95% CI=5.6-35.2), respectively. A sequencing analysis of the pCS20 gene of E. ruminantium revealed an E. ruminantium-like organism (1/849, 0.1%, 95% CI=0-0.3) in one tick DNA sample extracted from Rhipicephalus (Boophilus) microplus in Hunan. In addition, Borrelia americana in Ixodes persulcatus, Babesia occultans in Haemaphysalis qinghaiensis and both Rhipicephalus sanguineus and an Ehrlichia muris-like organism in R. (B.) microplus was detected, possibly for the first time in China. Four DNA sequences closely related to Borrelia carolinensis and/or Borrelia bissettii from Haemaphysalis longicornis, Candidatus Rickettsia principis from H. qinghaiensis, and I. persulcatus and Ehrlichia canis (named E. canis-like) from Haemaphysalis bispinosa were also detected in this work. Copyright © 2016 Elsevier B.V. All rights reserved.

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

    PubMed

    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.

  15. 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

  16. 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

  17. 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

  18. 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.

  19. Evaluation of medicinal plant extracts against ticks and fluke.

    PubMed

    Elango, Gandhi; Rahuman, Abdul Abdul

    2011-03-01

    The present study was based on assessments of the antiparasitic activities to determine the efficacies of leaf hexane, chloroform, ethyl acetate, acetone and methanol extracts of Aegle marmelos (Linn.) Correa ex Roxb, Andrographis lineata Wallich ex Nees., Andrographis paniculata (Burm.f.) Wallich ex Nees., Cocculus hirsutus (L.) Diels, Eclipta prostrata L., and Tagetes erecta L. against the adult cattle tick Haemaphysalis bispinosa Neumann 1897 (Acarina: Ixodidae), the larvae of Rhipicephalus (Boophilus) microplus Canestrini 1887 (Acari: Ixodidae) and sheep fluke Paramphistomum cervi Zeder 1790 (Digenea: Paramphistomatidae). All plant extracts showed moderate toxic effect on parasites after 24 h of exposure; however, the highest parasitic activity was found in leaf ethyl acetate extract of A. lineata, methanol extract of A. marmelos, A. paniculata, and C. hirsutus against H. bispinosa (LC(50) = 395.27, 358.45, 327.21 and 420.50 ppm); ethyl acetate extract of A. paniculata, C. hirsutus, methanol extracts of A. marmelos, A. lineata, and E. prostrata against the larvae of R. microplus (LC(50) = 207.70, 258.61, 134.09, 206.00, and 274.33 ppm); hexane extract of A. lineata, ethyl acetate extract of A. paniculata, E. prostrata, acetone extracts of T. erecta, methanol extracts of A. marmelos and C. hirsutus against P. cervi (LC(50) = 254.23, 451.17, 425.73, 253.60, 542.71, and 360.17 ppm), respectively. The present study is the first report on the veterinary parasitic activity of plant extracts from Southern India.

  20. 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.

  1. Tick fauna from two locations in the Brazilian savannah.

    PubMed

    Szabó, Matias Pablo Juan; Olegário, Maria Marlene Martins; Santos, André Luiz Quagliatto

    2007-01-01

    The Cerrado is Brazil's tropical savannah, which is arguably under greater threat than the Amazon rainforest. The Cerrado Biome of tropical South America covers about 2 million km(2) and is considered a biodiversity hot spot which means that it is especially rich in endemic species and particularly threatened by human activities. The Cerrado is increasingly exposed to agricultural activities which enhance the likelihood of mixing parasites from rural, urban and wildlife areas. Information about ticks from the Cerrado biome is scarce. In this report tick species free-living, on domestic animals and on a few wild animals in two farms in the Cerrado biome (Nova Crixás and Araguapaz municipalities, Goiás State, Brazil) are described. Amblyomma cajennense was the first and Amblyomma parvum the second host-seeking tick species found. Only two other tick species were found free-living: one Amblyomma nodosum and three Amblyomma naponense nymphs. Cattle were infested with Boophilus microplus and A. cajennense. Buffalos were infested with B. microplus and A. parvum. Dogs were infested with A. cajennense, Amblyomma ovale, A. parvum and Rhipicephalus sanguineus ticks. Anocentor nitens, B. microplus, A. cajennense, and A. parvum were found on horses. Amblyomma auricularium were found attached to nine-banded armadillos and Amblyomma rotundatum to red-footed tortoise, cururu toads and a rattlesnake. The latter was also infested with an adult A. cajennense. No tick was found on a goat, a tropical rat snake and a yellow armadillo. Among the observations the infestation of several domestic animals with A. parvum seems be the main feature. It suggests that this species might become a pest. However, the life cycle of A. parvum in nature, as well as its disease vectoring capacity, are largely unknown. It would be important to determine if it is a species expanding its geographic range by adaptation to new hosts or if it has been maintained in high numbers at definite locations by

  2. 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

  3. 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.

  4. The vulnerability of animal and human health to parasites under global change.

    PubMed

    Sutherst, R W

    2001-07-01

    The term 'global change' is used to encompass all of the significant drivers of environmental change as experienced by hosts, parasites and parasite managers. The term includes changes in climate and climate variability, atmospheric composition, land use and land cover including deforestation and urbanisation, bio-geochemistry, globalisation of trade and transport, the spread of alien species, human health and technology. A subset of land use issues relates to the management of protective technologies in relation to residues in food and the environment and the emergence of resistance. Another is the question of changing biodiversity of both parasites and their associated natural enemies, and the effects on the host--parasite relationship and on parasite management. A framework for studying impacts of global change is proposed and illustrated with field data, and CLIMEX and simulation modelling of the cattle tick Boophilus microplus in Australia. Parasitology suffers from the perception that the key impacts of global change will be driven by changes at lower trophic levels, with parasitic interactions being treated as secondary effects. This is incorrect because the environment mediates host-parasite interactions as much as it affects parasites directly. Parasitologists need to strive for holistic solutions to the management of animal and human health, within a wider context of overall management of those systems, if they are to make a meaningful contribution to global efforts aimed at coping with global change.

  5. [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).

  6. [Grazing systems, rotenone and parasites control in crossbred calves: effect on live weight gain and on parasites burdens].

    PubMed

    Catto, João B; Bianchin, Ivo; Santurio, Jânio M; Feijó, Gelson L D; Kichel, Armindo N; Silva, José M da

    2009-01-01

    Practices for endo and ectoparasite control in beef cattle were evaluated in two independent experiments. First, the effects of rotenone on Rhipicephalus (Boophilus) microplus ticks were evaluated in vitro and in experimentally infected calves. In the second trial, the effects of grazing systems associated with endo and ectoparasite treatments on parasite burden and weight gain of naturally parasited animals were evaluated. Rotenone showed acaricide action on larvae and engorged ticks during in vitro tests and on larvae in experimentally infected calves. Three treatments with endectocide decreased (P < .05) the number of EPG and ticks and increased (P < .05) the weight gain in the dry season. Animals treated with only one application of levamisole showed EPG intermediate and different (P < .05) from the groups treated with endectocide (lower) and control (higher) in the dry season, but the weight gain obtained with this treatment did not differ from the control group. During the raining season the animals treated with fipronil were significantly less parasited by horn fly, tick and larvae of Dermatobia hominis and the group treated with rotenone were significantly less parasited by horn fly in relation to control. Animals under rotational grazing showed significantly higher EPG than those under continuous grazing. Three treatments with endectocide in the dry season plus three acaricide treatments with fipronil in the raining season reduced EPG, tick, and screw worm larva counts, and provided a significant increase (23 kg) of live weight gain in relation to untreated animals.

  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. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. Bovine babesiosis and anaplasmosis follow-up on cattle relocated in an endemic area for hemoparasitic diseases.

    PubMed

    Figueroa, J V; Alvarez, J A; Ramos, J A; Rojas, E E; Santiago, C; Mosqueda, J J; Vega, C A; Buening, G M

    1998-06-29

    A multiplex PCR/DNA probe assay was used to monitor Babesia bovis, B. bigemina and Anaplasma marginale infection in cattle introduced to a Boophilus microplus-infested area in Veracruz, Mexico. Eight intact, 18-month-old, cross-bred beef cattle (four naive, Group A; four Babesia species--premunized, Group B) were immediately exposed to ticks after arrival and were clinically monitored from day 6 to day 98 post-exposure (PE) to ticks. Blood sample analysis for DNA detection by the MPCR/DNA probe assay showed that Group A animals were infected with B. bovis from day 11 up to day 22 PE, requiring treatment on days 17-20. Group B animals were detected positive to B. bovis on days 17-20, did not require treatment and remained persistently infected from days 70 to 84 PE. Treatment of Group A animals delayed the infection with B. bigemina. These animals became positive to the parasite on days 63-77 PE. In contrast, Group B animals (untreated) showed B. bigemina infection on days 21-26 and 63-84 PE. One animal was positive for A. marginale infection on days 63-66 PE, the rest of the animals became so on days 80-98 PE. All infected animals required treatment with oxytetracycline. Monitoring the triple hemoparasite infection with the MPCR/DNA probe assay provided important epidemiological information. Thus, precautionary measures can be established when cattle are moved to a babesiosis/anaplasmosis risk area.

  18. Clinicopatho-Biochemical Alterations Associated with Subclinical Babesiosis in Dairy Animals

    PubMed Central

    Sharma, Amrita; Singla, Lachhman Das; Ashuma; Batth, Balvinder Kaur; Kaur, Paramjit

    2016-01-01

    Background: Present investigation is based on the clinicopatho-biochemical alteration related to natural tick borne bovine babesiosis in Punjab state. Methods: Blood samples from jugular vein of 542 bovines (cattle 466 and buffaloes 76) having history of tick infestation, fever, hemoglobinurea or anemia were collected and tested for Babesia bigemina by blood smear examination and PCR targeting 18S rRNA gene to distinguish clinically and subclinically infected groups. Further the hemato-biochemical parameters were correlated with the status of infection. Results: Overall, of the 542 samples tested 16.42 % were positive by PCR while only 1.66 % by blood smear examination. The trend of molecular prevalence was found to decrease from north-eastern towards western Punjab. Analysis of the hematobiochemical alterations showed significant decrease in the levels of RBC, Hb, PCV, and MCV with significant increase in TBIL, MCH and MCHC levels. Conclusion: As the transmission of B. bigemina is transovarian, presence of even few infected Rhipicephalus (Boophilus) microplus ticks on a subclinically infection can be the nidus of infection for whole herd, causing severe economic losses, at the same time significantly affecting the physiology of carrier animal. PMID:27308284

  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. 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

  1. 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

  2. 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.

  3. 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.

  4. Ticks and tick-borne disease in Guatemalan cattle and horses.

    PubMed

    Teglas, Mike; Matern, Erin; Lein, Sarah; Foley, Patrick; Mahan, Suman M; Foley, Janet

    2005-07-15

    Blood samples and ticks were collected from 48 cattle and 74 horses from seven sites in the Peten region of Guatemala. Data on body condition, mucous membrane capillary refill time and tick infestation levels were recorded for each animal in the study. Horses had significantly higher levels of tick infestation than cattle, as well as poorer body condition scores. Seroprevalence of Babesia spp. was 95.8% for B. bovis in cattle, 89.6% for B. bigemina in cattle, and 92.7% for B. equi in horses. Seroprevalence of Anaplasma marginale in cattle was 87.5%, similar to reports in animals from other regions of Central America. This is the first time that A. phagocytophilum has been reported in animals from this region, with overall PCR-prevalence of 27.6% in cattle and horses, and seroprevalence of 28.4% (52% in cattle and 13% in horses). An agent was identified with serological cross-reactivity and close genetic relatedness to Ehrlichia ruminantium, but further testing confirmed that the agent in Guatemalan cows was not the agent of heartwater. Ticks were identified to species with the predominant species identified on cattle as Boophilus microplus and Amblyomma cajennense, while Anocentor nitens and A. cajennense were most commonly found on horses. Prevalence of infection, tick infestation levels, host factors and environmental data were analyzed for association; A. nitens was significantly associated with A. phagocytophilum prevalence by village.

  5. Exposed and concealed antigens as vaccine targets for controlling ticks and tick-borne diseases.

    PubMed

    Nuttall, P A; Trimnell, A R; Kazimirova, M; Labuda, M

    2006-04-01

    Tick vaccines derived from Bm86, a midgut membrane-bound protein of the cattle tick, Boophilus microplus, are currently the only commercially available ectoparasite vaccines. Despite its introduction to the market in 1994, and the recognized need for alternatives to chemical pesticides, progress in developing effective antitick vaccines (and ectoparasite vaccines in general) is slow. The primary rate-limiting step is the identification of suitable antigenic targets for vaccine development. Two sources of candidate vaccine antigens have been identified: 'exposed' antigens that are secreted in tick saliva during attachment and feeding on a host and 'concealed' antigens that are normally hidden from the host. Recently, a third group of antigens has been distinguished that combines the properties of both exposed and concealed antigens. This latter group offers the prospect of a broad-spectrum vaccine effective against both adults and immature stages of a wide variety of tick species. It also shows transmission-blocking and protective activity against a tick-borne pathogen. With the proliferation of molecular techniques and their application to vaccine development, there are high hopes for new and effective antitick vaccines that also control tick-borne diseases.

  6. 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.

  7. 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.

  8. 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

  9. Isolation and characterisation of acaricidal and larvicidal novel compound (2S,5R,6R)-2-hydroxy-3,5,6-trimethyloctan-4-one from Streptomyces sp. against blood-sucking parasites.

    PubMed

    Deepika, Thimiri Lakshmipathi; Kannabiran, Krishnan; Khanna, Venkatesan Gopiesh; Rajakumar, Govindasamy; Jayaseelan, Chidambaram; Santhoshkumar, Thirunavukkarasu; Rahuman, Abdul Abdul

    2012-09-01

    The aim of the present study was to assess the acaricidal and larvicidal property of marine actinobacterial compound (2S,5R,6R)-2-hydroxy-3,5,6-trimethyloctan-4-one extracted and isolated from Streptomyces sp. VITDDK3 tested against the larvae of Rhipicephalus (Boophilus) microplus Canestrini (Acari: Ixodidae), Anopheles subpictus Grassi and Culex quinquefasciatus Say (Diptera: Culicidae). The isolate was taxonomically characterised, identified and designated as Streptomyces sp. VITDDK3. The crude compound was loaded on silica gel column and eluted with chloroform-methanol-water. The purity of the compound isolated was analysed by thin layer chromatography using chloroform and methanol as the solvent system and confirmed by high-performance liquid chromatography. The structure of the purified compound was established from infrared, ultraviolet, (1)H-nuclear magnetic resonance (NMR), (13)C-NMR and mass spectral data. The chemical shift assignments obtained for the aliphatic compound from (1)H-NMR corresponding to the molecular formula C(11)H(22)O(2). Bioassay-guided fractionation led to the isolation of compound, which was identified as (2S,5R,6R)-2-hydroxy-3,5,6-trimethyloctan-4-one. In the present study, Streptomyces sp. VITDDK3 crude extract and different fractions were tested against the larvae of parasites at a concentration of 1,000 ppm. Those fractions showing 100% mortality in 24 h alone was selected for further column chromatographic separation. The purified compound (2S,5R,6R)-2-hydroxy-3,5,6-trimethyloctan-4-one was tested in the concentrations of 250, 200, 150, 100 and 50 ppm and observed the per cent mortality of 100, 88, 62, 50 and 36 against R. microplus; 100, 100, 87, 62 and 39 against A. subpitcus; and 100, 94, 79, 51 and 33 against C. quinquefasciatus, respectively. The crude extract showed parasitic effects after 24 h of exposure at 1,000 ppm, and parasite mortality was observed against the larvae of R. microplus (LC(50) = 88.74 ppm; r (2) = 0

  10. Immunogenic potential of the recombinant Rhipicephalus microplus aquaporin protein against the tick Rhipicephalus sanguineus Latreille, 1806 in domestic dogs

    USDA-ARS?s Scientific Manuscript database

    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...

  11. 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.

  12. 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.

  13. 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

  14. 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

  15. 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).

  16. 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

  17. Eco-friendly approach using marine actinobacteria and its compounds to control ticks and mosquitoes.

    PubMed

    Thenmozhi, Mohankumar; Gopal, Jannu Vinay; Kannabiran, Krishnan; Rajakumar, Govindasamy; Velayutham, Kanayairam; Rahuman, Abdul Abdul

    2013-02-01

    Ticks and mosquitoes are ectoparasitic arthropods that can transmit a variety of diseases to humans and animals during blood feeding and causing serious infectious disorders. The purpose of the present study was to assess the acaricidal and insecticidal property of ethyl acetate extract and its compounds isolated from marine actinobacteria, Streptomyces VITSTK7 sp. against the larvae of cattle ticks, Haemaphysalis bispinosa and Rhipicephalus (Boophilus) microplus (Acari: Ixodidae); fourth-instar larvae of malaria vector, Anopheles subpictus; and filarial vector, Culex quinquefasciatus (Diptera: Culicidae). The ethyl acetate extract was loaded on silica gel column and separated with chloroform, methanol, and acetone as the solvents system. The separation of fractions was visualized by the thin layer chromatography (TLC) plate, further confirmed by high-performance liquid chromatography, and followed by gas liquid chromatography. Three major fractions were analyzed in mass spectroscopy (MS) and matched with existing compounds in the data base. Based on the fragment pattern, it led to the major compounds which were predicted as cyclopentanepropanoic acid, 3,5-bis(acetyloxy)-2-[3-(methoxyimino)octyl], methyl ester (13.3 %) 1; 5-azidomethyl-3-(2-ethoxy carbonyl-ethyl)-4-ethoxycarbonylmethyl-1H-pyrrole-2-carboxylic acid, ethyl ester (18.2 %) 2; and akuammilan-16-carboxylic acid, 17-(acetyloxy)-10-methoxy, methyl ester (16R) (53.3 %) 3. The maximum efficacy was observed in compounds 1, 2, and 3, and the ethyl acetate extract of Streptomyces VITSTK7 sp. against the larvae of H. bispinosa (LC(50) = 1,573.36, 1,333.09, 1,073.29, and 409.71 ppm; r(2) = 0.0.990, 0.934, 0.935, and 0.908), R. microplus (LC(50) = 1,877.86, 815.83, 1,631.14, and 441.54 ppm; r(2) = 0.981, 0.926, 0.0970, and 0.915), A. subpictus (LC(50) = 273.89, 687.69, 464.75, and 223.83 ppm; r(2) = 0.758, 0.924, 0.841, and 0.902), and C. quinquefasciatus (LC(50) = 430.06, 881.59, 777.0, and 195.70 ppm; r(2) = 0

  18. Prevalence and molecular heterogeneity of Bartonella bovis in cattle and Haemaphysalis bispinosa ticks in Peninsular Malaysia.

    PubMed

    Kho, Kai-Ling; Koh, Fui-Xian; Jaafar, Tariq; Nizam, Quaza Nizamuddin Hassan; Tay, Sun-Tee

    2015-07-16

    Bartonellosis is an emerging zoonotic infection responsible for a variety of clinical syndromes in humans and animals. Members of the genus Bartonella exhibit high degrees of genetic diversity and ecologic plasticity. The infection is usually transmitted to animals and humans through blood-feeding arthropod vectors such as fleas, lice, ticks and sandflies. This study was conducted to investigate the prevalence of Bartonella species in 184 beef cattle, 40 dairy cattle, 40 sheep and 40 goats in eight animal farms across Peninsular Malaysia. Bartonella-specific PCR assays and sequence analysis of partial fragments of the citrate synthase gene were used for detection and identification of B. bovis. Isolation of B. bovis was attempted from PCR-positive blood samples. Molecular heterogeneity of the isolates was investigated based on sequence analysis of gltA, ITS, rpoB genes, ERIC-PCR, as well as using an established multilocus sequence typing (MLST) method. The carriage rate of B. bovis in ticks was also determined in this study. B. bovis was detected using Bartonella gltA-PCR assays from ten (4.5 %) of 224 cattle blood samples, of which three (1.3 %) were from beef cattle and seven (3.1 %) were from dairy cattle. None of the blood samples from the sheep and goats understudied were positive for B. bovis. Haemaphysalis bispinosa and Rhipicephalus (Boophilus) microplus were the predominant tick species identified in this study. B. bovis was detected from eight of 200 H. bispinosa ticks and none from the R. microplus ticks. Isolation of B. bovis was successful from all PCR-positive cattle blood samples, except one. Strain differentiation of B. bovis isolates was attempted based on sequence analysis of gltA, ITS, rpoB, and ERIC-PCR assay. B. bovis isolates were differentiated into six genotypes using the approach. The genetic heterogeneity of the isolates was confirmed using MLST method. Of the six MLST sequence types identified, five were designated new sequence types (ST

  19. 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, respe