203 research outputs found

    Application of RNA interference to studies on biology of termites

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    The RNA interference (RNAi) mechanism can be used to reduce the expression of a selected gene in an organism. This method, discovered in 1998, has become the "gold standard" in basic biological research with overlaps to applied research and gene therapy in human medicine. For many reasons, RNAi is a suitable tool for the studies on biology of insects. RNAi is endowed with high sequence specificity, low costs, and easy application also in non-model species. Termites (Isoptera) are very sensitive to RNAi and this method is widely used to understand their physiology and ontogeny. The use of RNAi also has a potential in applied termite research and a significant number of publications have focused on the development of RNAi techniques as non-chemical pesticides against economically important termite species. This bachelor thesis aims to give a broad overview of the existing research on termites that uses the RNAi method.Mechanismus RNA interference (RNAi) lze vyuŇĺ√≠t ke sn√≠Ňĺen√≠ exprese vybran√©ho genu v organismu. Tato metoda, objeven√° v roce 1998, se stala "zlat√Ĺm standardem" v z√°kladn√≠m biologick√©m v√Ĺzkumu s pŇôesahem do v√Ĺzkumu aplikovan√©ho a genov√Ĺch terapi√≠ v medic√≠nńõ. Z mnoha dŇĮvodŇĮ je vhodn√Ĺm n√°strojem tak√© pro studium biologie hmyzu. Mezi hlavn√≠ patŇô√≠ vysok√° sekvenńćn√≠ specifita, n√≠zk√© n√°klady a nekomplikovan√© pouŇĺit√≠ i u nemodelov√Ĺch druhŇĮ. Termiti (Isoptera) jsou vŇĮńći RNAi vysoce citliv√≠ a tato metoda je Ň°iroce vyuŇĺ√≠v√°na k pozn√°v√°n√≠ jejich fyziologie a ontogeneze. VyuŇĺit√≠ RNAi m√° velk√Ĺ potenci√°l i v r√°mci aplikovan√©ho v√Ĺzkumu termitŇĮ a nemal√° ńć√°st publikac√≠ se tak zamńõŇôuje na v√Ĺvoj RNAi technik jako nechemick√Ĺch pesticidŇĮ proti ekonomicky v√Ĺznamn√Ĺm termit√≠m druhŇĮm. Tato bakal√°Ňôsk√° pr√°ce si klade za c√≠l podat Ň°irok√Ĺ pŇôehled dosavadn√≠ho v√Ĺzkumu vyuŇĺ√≠vaj√≠c√≠ metodu RNAi pŇôi studiu termitŇĮ.Katedra zoologieDepartment of ZoologyFaculty of SciencePŇô√≠rodovńõdeck√° fakult

    Molecular Survey of <i>Rickettsia raoultii</i> in Ticks Infesting Livestock from Pakistan with Notes on Pathogen Distribution in Palearctic and Oriental Regions

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    Ticks are hematophagous ectoparasites that transmit different pathogens such as Rickettsia spp. to domestic and wild animals as well as humans. Genetic characterizations of Rickettsia spp. from different regions of Pakistan are mostly based on one or two genetic markers and are confined to small sampling areas and limited host ranges. Therefore, this study aimed to molecularly screen and genetically characterize Rickettsia spp. in various tick species infesting camels, sheep, and goats. All the collected tick specimens were morphologically identified, and randomly selected tick species (148) were screened molecularly for the detection of Rickettsia spp. by amplifying three rickettsial DNA fragments, namely, the citrate-synthase gene (gltA), outer-membrane protein A (ompA), and outer-membrane protein B (ompB). After examining 261 hosts, 161 (61.7%) hosts were found infested by 564 ticks, including 287 (50.9%) nymphs, 171 (30.3%) females, and 106 (18.8%) males in five districts (Kohat, Dera Ismail Khan, Lower Dir, Bajaur, and Mansehra). The highest occurrence was noted for Hyalomma dromedarii (number = 72, 12.8%), followed by Haemaphysalis sulcata (n = 70, 12.4%), Rhipicephalus turanicus (n = 64, 11.3%), Rhipicephalus microplus (n = 55, 9.7%), Haemaphysalis cornupunctata (n = 49, 8.7%), Hyalomma turanicum (n = 48, 8.5%), Hyalomma isaaci (n = 45, 8.0%), Haemaphysalis montgomeryi (n = 44, 7.8%), Hyalomma anatolicum (n = 42, 7.5%), Haemaphysalis bispinosa (n = 38, 6.7%), and Rhipicephalus haemaphysaloides (n = 37, 6.6%). A subset of 148 ticks were tested, in which eight (5.4%) ticks, including four Hy. turanicum, two Ha. cornupunctata, one Ha. montgomeryi, and one Ha. bispinosa, were found positive for Rickettsia sp. The gltA, ompA, and ompB sequences revealed 100% identity and were phylogenetically clustered with Rickettsia raoultii reported in China, Russia, USA, Turkey, Denmark, Austria, Italy, and France. Additionally, various reports on R. raoultii from Palearctic and Oriental regions were summarized in this study. To the best of our knowledge, this is the first report regarding genetic characterization and phylogenetic analysis of R. raoultii from Pakistan. Further studies to investigate the association between Rickettsia spp. and ticks should be encouraged to apprise effective management of zoonotic consequences

    Molecular detection of Rickettsia hoogstraalii in Hyalomma anatolicum and Haemaphysalis sulcata : updated knowledge on the epidemiology of tick-borne Rickettsia hoogstraalii

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    Ticks are hematophagous ectoparasites that transmit pathogens to animals and humans. Updated knowledge regarding the global epidemiology of tick-borne Rickettsia hoogstraalii is dispersed, and its molecular detection and genetic characterization are missing in Pakistan. The current study objectives were to molecularly detect and genetically characterize Rickettsia species, especially R. hoogstraalii, in hard ticks infesting livestock in Pakistan, and to provide updated knowledge regarding their global epidemiology. Ticks were collected from livestock, including goats, sheep, and cattle, in six districts of Khyber Pakhtunkhwa (KP) Pakistan. Overall, 183 hosts were examined, of which 134 (73.2%), including goats (number = 39/54, 72.2%), sheep (23/40, 57.5%), and cattle (71/89, 80%) were infested by 823 ticks. The most prevalent tick species was Rhipicephalus microplus (number = 283, 34.3%), followed by Hyalomma anatolicum (223, 27.0%), Rhipicephalus turanicus (122, 14.8%), Haemaphysalis sulcata (104, 12.6%), Haemaphysalis montgomeryi (66, 8.0%), and Haemaphysalis bispinosa (25, 3.03%). A subset of 210 ticks was selected and screened for Rickettsia spp. using PCRbased amplification and subsequent sequencing of rickettsial gltA and ompB fragments. The overall occurrence rate of R. hoogstraalii was 4.3% (number = 9/210). The DNA of Rickettsia was detected in Hy. anatolicum (3/35, 8.5%) and Ha. sulcata (6/49, 12.2%). However, no rickettsial DNA was detected in Rh. microplus (35), Rh. turanicus (35), Ha. montgomeryi (42), and Ha. bispinosa (14). The gltA and ompB fragments showed 99‚Äď100% identity with R. hoogstraalii and clustered phylogenetically with the corresponding species from Pakistan, Italy, Georgia, and China. R. hoogstraalii was genetically characterized for the first time in Pakistan and Hy. anatolicum globally. Further studies should be encouraged to determine the role of ticks in the maintenance and transmission of R. hoogstraalii in different hosts

    Correlation between Babesia species affecting dogs in Taiwan and the local distribution of the vector ticks

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    DATA AVAILABILITY : All data are included in the current paper.The objective of our study was to survey Babesia infection rates by PCR and tick species on stray dogs to correlate the distribution of Babesia with the distribution of ticks infesting dogs in Taiwan. Three hundred eighty-eight blood samples and 3037 ticks were collected from 388 roaming, and free-ranging owned dogs at residential sites in Taiwan between January 2015 and December 2017. The prevalence of B. gibsoni and B. vogeli was 15.7% (61/388) and 9.5% (37/388), respectively. Most positive B. gibsoni dogs were found in the northern part of the country 56/61 (91.8%), whereas a few were found in the middle 5/61 (8.2%). Babesia vogeli infection rates were 10%, 3.6%, and 18.2% in the northern, central, and southern regions, respectively. Five species of ticks were found: Rhipicephalus sanguineus (throughout Taiwan), Rhipicephalus haemaphysaloides (in the north), Haemaphysalis hystricis (in the north and middle of Taiwan), and Amblyomma testidunarium and Ixodes ovatus (both in the north). None of the dogs in the south were infected with B gibsoni, which correlated with the absence of H. hystricis, a tick recently identified as the local vector for B gibsoni. Babesia vogeli was more equally distributed, coinciding with R. sanguineus, a tick that is present throughout Taiwan. Anaemia was detected in 86.9% of infected dogs; among these dogs, approximately 19.7% showed severe anaemia (HCT < 20). These findings provide useful advice for owners regarding outdoor activities with their dogs and local veterinarians with a regional differential diagnosis of babesiosis in Taiwan.https://www.mdpi.com/journal/vetsciVeterinary Tropical Disease

    The ecological and etiological investigation of ticks and rodents in China: results from an ongoing surveillance study in Zhejiang Province

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    ObjectivesThis study aimed to analyze the population density of vector ticks and reservoir hosts rodents, and to investigate the relevant pathogen infection in Zhejiang Province, China.MethodsIn this surveillance study, the data of ticks density were collected with the tick picking method on animal body surface and the drag-flag method, while the rodent density with the night trapping method. The samples of ticks were examined for the severe fever with thrombocytopenia syndrome virus (SFTSV), and blood serum and organs from rodents were subjected for SFTSV, hantavirus, Leptospira, Orientia tsutsugamushi (O. tsutsugamushi) and Yersinia pestis (Y. pestis) screening in the laboratory.ResultsFrom 2017 to 2022 in Zhejiang Province, 16,230 parasitic ticks were found in 1848 positive animals, with the density of parasitic ticks of 1.29 ticks per host animal, and a total of 5,201 questing ticks were captured from 1,140,910 meters of vegetation distance with the questing tick density of 0.46 ticks/flag¬∑100‚ÄČm. Haemaphysalis longicornis (H. longicornis) was the major species. A total of 2,187,739 mousetraps were distributed and 12,705 rodents were trapped, with the density of 0.58 per 100 trap-nights. Rattus norvegicus was the major species. For SFTSV screening, two groups nymphal ticks of H. longicornis were tested to be positive. For the rodents samples, the Leptospira had a positive rate of 12.28% (197/1604), the hantavirus was 1.00% (16/1604), and the O. tsutsugamushi was 0.15% (2/1332). No positive results were found with SFTSV and Y. pestis in the rodents samples.ConclusionFindings from this study indicated that the ticks and rodents were widely distributed in Zhejiang Province. Particularly, the positive detection of SFTSV, Leptospira, hantavirus and O. tsutsugamushi in ticks or rodents from this area suggested that more attention should be paid to the possibilities of relevant vector-borne diseases occurrence

    Efficacy of the Vaccine Candidate Based on the P0 Peptide against Dermacentor nitens and Ixodes ricinus Ticks

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    (This article belongs to the Collection Advances in Tick Research)The control of ticks through vaccination offers a sustainable alternative to the use of chemicals that cause contamination and the selection of resistant tick strains. However, only a limited number of anti-tick vaccines have reached commercial realization. In this sense, an antigen effective against different tick species is a desirable target for developing such vaccines. A peptide derived from the tick P0 protein (pP0) conjugated to a carrier protein has been demonstrated to be effective against the Rhipicephalus microplus, Rhipicephalus sanguineus, and Amblyomma mixtum tick species. The aim of this work was to assess the efficacy of this peptide when conjugated to the Bm86 protein against Dermacentor nitens and Ixodes ricinus ticks. An RNAi experiment using P0 dsRNA from I. ricinus showed a dramatic reduction in the feeding of injected female ticks on guinea pigs. In the follow-up vaccination experiments, rabbits were immunized with the pP0-Bm86 conjugate and challenged simultaneously with larvae, nymphs, and the adults of I. ricinus ticks. In the same way, horses were immunized with the pP0-Bm86 conjugate and challenged with D. nitens larva. The pP0-Bm86 conjugate showed efficacies of 63% and 55% against I. ricinus and D. nitens ticks, respectively. These results, combined with previous reports of efficacy for this conjugate, show the promising potential for its development as a broad-spectrum anti-tick vaccine.This research was funded by the Center for Genetic Engineering and Biotechnology, Havana, Cuba, the Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Portugal, and the Czech Science Foundation grant no. 20-05736S. Mobility was supported by the CYTED Network INCOGARR 110RT0541.info:eu-repo/semantics/publishedVersio

    Association of SFG <i>Rickettsia massiliae</i> and <i>Candidatus</i> Rickettsia shennongii with Different Hard Ticks Infesting Livestock Hosts

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    Ixodid ticks are responsible for the transmission of various intracellular bacteria, such as the Rickettsia species. Little Information is available about the genetic characterization and epidemiology of Rickettsia spp. The current study was designed to assess the tick species infesting various livestock hosts and the associated Rickettsia spp. in Pakistan. Ticks were collected from different livestock hosts (equids, cattle, buffaloes, sheep, goats, and camels); morphologically identified; and screened for the genetic characterization of Rickettsia spp. by the amplification of partial fragments of the gltA, ompA and ompB genes. Altogether, 707 ticks were collected from 373 infested hosts out of 575 observed hosts. The infested hosts comprised 105 cattle, 71 buffaloes, 70 sheep, 60 goats, 34 camels, and 33 equids. The overall occurrence of Rickettsia spp. was 7.6% (25/330) in the tested ticks. Rickettsia DNA was detected in Rhipicephalus haemaphysaloides (9/50, 18.0%), followed by Rhipicephalus turanicus (13/99, 13.1%), Haemaphysalis cornupunctata (1/18, 5.5%), and Rhipicephalus microplus (2/49, 4.1%); however, no rickettsial DNA was detected in Hyalomma anatolicum (71), Hyalomma dromedarii (35), and Haemaphysalis sulcata (8). Two Rickettsia agents were identified based on partial gltA, ompA, and ompB DNA sequences. The Rickettsia species detected in Rh. haemaphysaloides, Rh. turanicus, and Rh. microplus showed 99‚Äď100% identity with Rickettsia sp. and Candidatus Rickettsia shennongii, and in the phylogenetic trees clustered with the corresponding Rickettsia spp. The Rickettsia species detected in Rh. haemaphysaloides, Rh. turanicus, Rh. microplus, and Ha. cornupunctata showed 100% identity with R. massiliae, and in the phylogenetic trees it was clustered with the same species. Candidatus R. shennongii was characterized for the first time in Rh. haemaphysaloides, Rh. turanicus, and Rh. microplus. The presence of SFG Rickettsia spp., including the human pathogen R. massiliae, indicates a zoonotic risk in the study region, thus stressing the need for regular surveillance

    Universal tick vaccines : candidates and remaining challenges

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    Recent advancements in molecular biology, particularly regarding massively parallel sequencing technologies, have enabled scientists to gain more insight into the physiology of ticks. While there has been progress in identifying tick proteins and the pathways they are involved in, the specificities of tick-host interaction at the molecular level are not yet fully understood. Indeed, the development of effective commercial tick vaccines has been slower than expected. While omics studies have pointed to some potential vaccine immunogens, selecting suitable antigens for a multiantigenic vaccine is very complex due to the participation of redundant molecules in biological pathways. The expansion of ticks and their pathogens into new territories and exposure to new hosts makes it necessary to evaluate vaccine efficacy in unusual and non-domestic host species. This situation makes ticks and tick-borne diseases an increasing threat to animal and human health globally, demanding an urgent availability of vaccines against multiple tick species and their pathogens. This review discusses the challenges and advancements in the search for universal tick vaccines, including promising new antigen candidates, and indicates future directions in this crucial research field

    TickProVac: An insight into the Rhipicephalus sanguineus tick salivary gland proteome during Ehrlichia canis infection towards vaccine development

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    As carra√ßas s√£o reconhecidas como vectores de diversos agentes patog√©nicos respons√°veis por doen√ßas relevantes na medicina humana e animal. Erliquiose monoc√≠tica canina (EMC), causada pela bact√©ria Ehrlichia canis, √© transmitida pela carra√ßa Rhipicephalus sanguineus. O potencial zoon√≥tico da EMC est√° bem documentado com relatos de infec√ß√Ķes humanas em aumento permanente. A infesta√ß√£o por carra√ßas e as doen√ßas por elas transmitidas (DTCs) continuam a representar um s√©rio problema devido √† falta de profilaxias eficazes. √Č, portanto, imperativo adotar novas abordagens, nomeadamente a utiliza√ß√£o de vacinas, para a redu√ß√£o de infesta√ß√Ķes por carra√ßas e DTCs. Actualmente n√£o existe vacina anti-R. sanguineus comercialmente dispon√≠vel. De forma a solucionar esta lacuna, o objetivo principal do presente trabalho foi a caracteriza√ß√£o de genes e prote√≠nas que atuam na interface vector-agente patog√©nico na presen√ßa de infe√ß√£o para compreender os mecanismos associados √† transmiss√£o de DTCs. Neste trabalho est√£o reportados genes e prote√≠nas diferencialmente expressos obtidos a partir da an√°lise transcript√≥mica e prote√≥mica das gl√Ęndulas salivares (GSs) de R. sanguineus durante a infe√ß√£o com E. canis. Os n√≠veis de express√£o dos genes que codificam as prote√≠nas serina carboxipeptidase (psc), prote√≠na de choque t√©rmico (phsrp20) e proibitina (prohib) foram investigados in vitro e in vivo, em amostras infectadas e n√£o infectadas. Posteriormente, o silenciamento de genes foi realizado atrav√©s de RNA de interfer√™ncia (RNAi) para avaliar o efeito da sub-express√£o de ferritina 1 na alimenta√ß√£o, desenvolvimento ov√°rico, oog√©nese e aquisi√ß√£o E. canis pela carra√ßa. O mesmo foi efetuado para determinar o efeito da sub-express√£o de psc, prohib e phsrp20 na invas√£o e multiplica√ß√£o desta bact√©ria na linha celular IDE8 e em R. sanguineus. O p√©ptido pPHSRP20, relacionado com o choque t√©rmico, foi sintetizado para avaliar a sua imunogenicidade em ratos CD1. Os resultados mostraram que o silenciamento de ferritina 1 compromete a compet√™ncia da carra√ßa em se alimentar e provoca altera√ß√Ķes morfol√≥gicas e histoqu√≠micas nos ov√°rios e o√≥citos. Os dados transcript√≥micos e prote√≥micos demonstraram que as altera√ß√Ķes da express√£o g√©nica e proteica est√£o principalmente relacionadas com processos proteicos celulares e metab√≥licos e com atividades catal√≠ticas, muito provavelmente um reflexo de altera√ß√Ķes de transcri√ß√£o e tradu√ß√£o em resposta √† presen√ßa de infec√ß√£o. Em amostras infectadas verificou-se uma sub-express√£o de phsrp20 e prohib em c√©lulas IDE8 e de phsrp20 e psc nas GSs. Inversamente, os genes psc nas c√©lulas IDE8 e prohib nas GSs estavam sobre-expressos. Apesar de se terem atingido n√≠veis elevados de silenciamento, o seu efeito na biologia de E. canis n√£o foi determinado devido a limita√ß√Ķes moleculares. pPHSRP20 desencadeou uma resposta imunit√°ria detect√°vel e espec√≠fica nos ratos CD1. Em geral, os resultados demonstraram que a presen√ßa de E. canis nas GSs desencadeia respostas celulares para regula√ß√£o do stress, inflama√ß√£o e rearranjo do citoesqueleto. A modula√ß√£o desta maquinaria molecular pela bact√©ria poder√° ser uma estrat√©gia para lidar e escapar √† resposta imunit√°ria e para utilizar prote√≠nas do vector necess√°rias √† virul√™ncia. Atrav√©s destes resultados esperou identificar-se potenciais antig√©nios de carra√ßa que contribuam para o desenvolvimento de uma vacina anti-R. sanguineus e/ou bloqueadora da transmiss√£o de agentes patog√©nicos.Ticks are recognised as potent vectors of disease-causing pathogens of medical and veterinary importance. Ehrlichia canis, the causative agent of canine monocytic ehrlichiosis (CME), is transmitted by the brown dog tick Rhipicephalus sanguineus and is acknowledged as a highly infectious disease. The zoonotic potential of CME is widely recognised with reports of human infections steadily increasing. Tick infestation and tick-borne diseases (TBDs) remain a serious and persistent veterinary health problem, due to the lack of efficient control measures. It is therefore vital that novel approaches to tackle TBDs are pursued. Although vaccination to reduce tick infestation is recognised, no anti- R. sanguineus vaccine exists. To address this, we aimed to characterise the crucial gene and protein interactions at the vector-pathogen interface to gain a fundamental understanding of the interactions underpinning disease transmission. Here we report differentially expressed genes and proteins found either in the literature or found in our transcriptomic and proteomic data from R. sanguineus salivary glands (SGs) during E. canis infection. The mRNA expression levels of the putative serine carboxypeptidase (psc), heat-shock related protein (phsrp20) and prohibitin-like protein genes were investigated in vitro and in vivo, in infected and uninfected samples. RNA interference (RNAi) was carried out to determine the effect of ferritin 1 in tick feeding, ovary (OV) development, oogenesis, and pathogen acquisition. We also determined the effect of a downregulation of three selected genes or proteins from our omics data on E. canis invasion and multiplication in the IDE8 tick cell line and R. sanguineus ticks. We synthesised a heat-shock related protein (pPHSRP20) peptide to evaluate its immunogenicity in CD1 mice. Our results have shown that silencing ferritin 1 alters tick competence to normally engorge and causes morphological and histochemical changes in the OV and oocytes. Our transcriptomic and proteomic data has revealed alterations in gene and protein expression, mostly concerning protein cellular and metabolic processes and catalytic activities, perhaps related with transcriptional and translational responses to E. canis infection. When analysing the transcription levels in E. canis-infected samples, a downregulation was verified for phsrp20 and prohib in IDE8 cells and phsrp20 and psc in the SGs, whereas an upregulation was observed for psc in IDE8 cells and prohib for the SGs. Even with high levels of gene silencing, the effect of gene silencing in E. canis biology was not determined due to molecular limitations. pPHSRP20 triggered a detectable and specific immune response in mice. Overall, results show that the presence of E. canis in the SGs leads to regulation of stress response, inflammation and cytoskeletal rearrangement molecules. Modulation of tick molecular machinery is a coping strategy to evade the host immune response and to utilise its proteins for infectivity. From this, we expected to identify tick antigens that will direct the development of an anti-R. sanguineus and/or transmission-blocking vaccine

    Proteína Bm86 y su potencial uso como vacuna contra garrapatas en el ganado bovino. Revisión

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    Tick infestations represent a major difficulty for cattle production in tropical and subtropical countries around the world. The traditional ways to combat this pest are chemicals (acaricides) and integrated management of grasslands. Nevertheless, these methods have not been able to conclusively control the presence of this ectoparasite. This has led to the search for vaccines that allow the control of these parasites. Currently, there are immunogens containing the gut antigen Bm86 of Rhipicephalus microplus, which have proven to be an effective alternative, as they have reduced tick populations and the use of synthetic acaricides. However, the polymorphism of this locus in ticks from different geographical areas represents a limitation in its effectiveness. A significant number of studies have been conducted on the Bm86 gene as a vaccine antigen, so this paper presents an update on studies conducted with this antigen and its potential use as an immunological control to reduce tick infestations in cattle.Las infestaciones de garrapatas representan una gran dificultad para la producci√≥n de ganado bovino en pa√≠ses tropicales y subtropicales alrededor del mundo. Las formas tradicionales de combate a esta plaga son qu√≠micos (acaricidas) y manejo integral de pastizales. Sin embargo, estos m√©todos no han logrado controlar de manera contundente la presencia de este ectopar√°sito. Lo anterior ha llevado a la b√ļsqueda de vacunas que permitan el control de estos par√°sitos. Actualmente, se cuenta con inmun√≥genos que contienen el ant√≠geno intestinal Bm86 de Rhipicephalus microplus, los cuales han demostrado ser una alternativa efectiva, al reducir las poblaciones de garrapatas y el uso de acaricidas sint√©ticos. No obstante, el polimorfismo de este locus en garrapatas de distintas zonas geogr√°ficas representa una limitante en su efectividad. Se han realizado un n√ļmero importante de investigaciones acerca del gen Bm86 como ant√≠geno vacunal, por lo que, en este documento, se presenta una actualizaci√≥n de las investigaciones realizadas con este ant√≠geno y su potencial uso como control inmunol√≥gico para reducir las infestaciones de garrapatas en el ganado bovino
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