Microplus Infestation Research Articles

Temporal analysis of the bovine lymph node transcriptome during cattle tick (Rhipicephalus microplus) infestation

Livestock production is a fundamental source of revenue and nutrition, wherein cattle-farming constitutes one of the major agricultural industries. Vectors and vector-borne diseases constitute one of the major factors that decrease the livelihood of all farming communities, more so in resource-poor communities and developing countries. Understanding the immunological responses during tick infestation in cattle is instrumental in the development of novel and improved tick control strategies, such as vaccines. In this study, gene expression patterns were compared within the lymph nodes of three cattle breeds at different life stages of the cattle tick, Rhipicephalus microplus. For Bonsmara (5/8Bos taurus indicus × 3/8B. t. taurus) cattle specifically, some 183 genes were found to be differentially expressed within the lymph nodes during larval and adult tick feeding, relative to uninfested cattle. Overall, the data provides evidence for a transcriptional regulatory network that is activated during immature tick infestation, but is down-regulated towards basal transcriptional levels when adult ticks are feeding. Specific processes in the lymph nodes of Bonsmara cattle were found to be differentially regulated on a transcriptional level. These include: (1) Leukocyte recruitment to the lymph node via chemokines and chemotaxis, (2) Trans-endothelial and intranodal movement on the reticular network, (3) Active regulation of cellular transcription and translation in the lymph node (including leukocyte associated cellular regulatory networks) and (4) Chemokine receptors regulating the movement of cells out of the lymph node. This work provides a first transcriptome analysis of bovine lymph node responses in tick-infested cattle. Findings show a dynamic immune response to tick infestation for the Bonsmara cattle breed, and that suppression of the maturation of the cattle hosts' immunity is especially evident during the larval feeding stages.

Bovine immunisation with a recombinant peptide derived from synthetic SBm7462® (Bm86 epitope construct) immunogen for Rhipicephalus microplus control

Rhipicephalus microplusis the most important ectoparasite of livestock in tropical and subtropical areas around the world. Research focused on developing an efficient vaccine for cattle tick control is a high priority. The aim of this study was to evaluate the rSBm7462® peptide (Bm86-B and T cell epitopes) regarding its properties of immunogenicity, protective effect in cattle and efficacy against R. microplus. This peptide was produced by a fermentative process in the yeast culture system of Komagataella (Pichia) pastoris strain Km 71. The vaccination assay was conducted in a tick-free area using non-splenectomised Holstein Friesian calves, separated into immunisation and control groups. These animals individually received the recombinant peptide and the inoculum without peptide using saponin as an adjuvant at three time points. The calves were challenged 21 days after the last immunisation using 4500 larvae per animal. An indirect ELISA was used to identify the IgG kinetics of serum samples from the calves studied. The qPCR was performed to determine the cytokine gene expression from the total RNA of the cultured PBMCs. Histomorphometry of the germinal centres (GCs) was performed measuring slides with haematoxylin-eosin staining of surgically removed lymph nodes from immunised calves. The antibody response showed a significant induction of high-affinity IgGs in calves immunised with the recombinant peptide in comparison to the controls. The kinetics of antibodies in immunised calves showed a significant increase during the experiment. This increase in high-affinity IgGs correlated with a gradual increase of the GC diameter following each peptide vaccination. Cytokine expression profiles demonstrating an adaptive immune response in calves immunised with rSBm7462® confirmed the T-dependent response. Vaccine efficacy was calculated at 72.4 % following the analysis and fecundity of collected adult female ticks, compared between control and vaccinated groups. These findings demonstrate that this new recombinant peptide is an option for control of R. microplus infestations.

Landscape Ecology of Rhipicephalus (Boophilus) microplus (Ixodida: Ixodidae) Outbreaks in the South Texas Coastal Plain Wildlife Corridor Including Man-Made Barriers.

Landscape features and the ecology of suitable hosts influence the phenology of invasive tick species. The southern cattle fever tick, Rhipicephalus (Boophilus) microplus (Canestrini) (Ixodida: Ixodidae), vectors causal agents of babesiosis in cattle and it infests exotic, feral nilgai, Bosephalus tragocamelus Pallas, and indigenous white-tailed deer, Odocoilus virginianus (Zimmerman), on the South Texas coastal plain wildlife corridor. The corridor extends from the Mexico border to cattle ranches extending north from inside Willacy Co. Outbreaks of R. microplus infesting cattle and nondomesticated ungulate hosts since 2014 in the wildlife corridor have focused attention on host infestation management and, by extension, dispersal. However, there is a knowledge gap on the ecology of R. microplus outbreaks in the South Texas coastal plain wildlife corridor. Ixodid distribution on the wildlife corridor is strongly influenced by habitat salinity. Saline habitats, which constitute ≈25% of the wildlife corridor, harbor few ixodids because of occasional salt toxicity from hypersaline wind tides and infrequent storm surges, and from efficient egg predation by mud flat fiddler crabs, Uca rapax (Smith). Rhipicephalus microplus infestations on nilgai were more prevalent in part of the corridor with mixed low salinity and saline areas than in an area that is more extensively saline. The different levels of R. microplus infestation suggest that man-made barriers have created isolated areas where the ecology of R. microplus outbreaks involve infested nilgai. The possible utility of man-made barriers for R. microplus eradication in the lower part of the South Texas coastal plain wildlife corridor is discussed.

Vaccine efficacy of recombinant BmVDAC on Rhipicephalus microplus fed on Babesia bigemina-infected and uninfected cattle

Rhipicephalus microplus is the most widely distributed tick worldwide and causes significant economic losses in the livestock industry. It directly affects hosts (especially in large infestations) by feeding on blood and piercing the skin and indirectly affects hosts as a vector of pathogens that cause infectious diseases, such as bovine babesiosis. Current research on the control of ticks is focused on integrated tick control programmes, including vaccination treatment with acaricides and completely blocking pathogen transmission. Our previous studies showed that R. microplus VDAC (BmVDAC) expression is modulated by Babesia bigemina infection. VDAC is a mitochondrial protein with multiple functions in addition to its primary role as a central component of the apoptotic machinery. In this paper, we evaluated BmVDAC as an anti-tick vaccine and its capacity to block the infection of Babesia bigemina in ticks. Our results demonstrate that rBmVDAC is immunogenic and that antibodies specifically recognize the native protein from midguts of R. microplus. Immunization with rBmVDAC afforded an 82% efficacy against R. microplus infestation in the group of vaccinated cattle compared with the control group. In contrast, rBmVDAC showed a lower efficacy of 34% against tick infestation in cattle vaccinated with rBmVDAC, infested with R. microplus and infected with B. bigemina. The main effect on ticks fed in vaccinated and infected cattle was a 34% reduction in egg fertility (DF) compared to ticks fed on the control group. There was no reduction in the B. bigemina parasite levels of ticks fed on rBmVDAC-vaccinated cattle. These results suggest that the rBmVDAC protein could be tested as a vaccine for the control of tick infestation.

Economic efficiency of Rhipicephalus microplus control and effect on beef cattle performance in the Brazilian Cerrado.

The cattle tick Rhipicephalus microplus causes significant economic losses to cattle production systems and is a main barrier to the introduction of Bos taurus breeds and their crosses in Brazil. These breeds have the genetic potential to generate animals that are more productive, but they are also more susceptible to R. microplus. One of the alternatives for conventional tick control is the use of strategic control, aiming at delaying or even preventing the development of its resistance to acaricides. The present study aimed to evaluate the economic losses caused by tick infestation on the productive performance of two breeds of beef cattle and to evaluate the economic efficiency of tick strategic control and its impacts on beef cattle production systems. Animal weights were obtained from the literature and were used to calculate the weight loss in kilograms (kg) and the economic loss (US$) caused by R. microplus infestation. The cost/loss ratio of performing strategic control was also calculated. The data show that tick infestation causes economic losses to the breeds and groups of animals evaluated and reduces animal performance (weight loss). The results show a loss of US$34.61/animal in the backgrounding phase and US$7.97/animal in the finishing phase for Brangus animals and its crosses. In conclusion, the data show that strategic control is economically efficient for Brangus animals in the backgrounding phase, independent of the methods used for acaricide application.

Acaricidal efficacy of Calotropis procera (Asclepiadaceae) and Taraxacum officinale (Asteraceae) against Rhipicephalus microplus from Mardan, Pakistan.

Medicinal plants are used by traditional folk healers, modern physicians, and veterinarians as an alternative to conventional drugs to treat a wide range of disorders including parasitic diseases. Some compounds from these plants have been shown to have acaricidal activity and repel arthropods. The cattle tick Rhipicephalus microplus is one of the most destructive pests to the livestock industry in tropical and subtropical parts of the world. The potential to develop herbal acaricides to control R. microplus infestations is critical in maintaining cattle herd productivity, reducing economic losses, and curtailing the overuse of synthetic chemical acaricides. Calotropis procera, the apple of Sodom, and Taraxacum officinale, the common dandelion, were evaluated for acaricidal activity against R. microplus larvae and adults in vitro. Both plant species tested are common indigenous species of Pakistan where R. microplus infestations are widespread across livestock species including cattle, sheep, and goats. Whole-plant extracts derived from C. procera and T. officinale significantly reduced the index of egg laying (P < 0.01) and increased the percent inhibition of oviposition of adult female ticks at a concentration of 40mg/mL when assessed by the adult immersion test (AIT). Calotropis procera and T. officinale treatments at the same concentration also resulted in larval mortality of 96.0% ± 0.57 and 96.7% ± 0.88, respectively, as measured using the larval packet test (LPT). An increasing range of extract concentrations was tested to determine the LD50 and LD90 for C. procera, 3.21and 21.15mg/mL, respectively, and T. officinale, 4.04and 18.92mg/mL, respectively. These results indicate that further studies are warranted to determine the relative contribution of individual phytochemicals from whole-plant extracts on acaricidal activity. This information will guide the design of further acaricidal efficacy tests using livestock infested with R. microplus.

Distinct immune response profile during rhipicephalus (boophilus) microplus infestations of guzerat dairy herd according to the maternal lineage ancestry (mitochondrial DNA)

Rhipicephalus (Boophilus) microplus ticks cause major constraints to public and livestock health, and serious economic losses. It is well known that the immune response to infestations with cattle ticks is influenced by the host genetic background leading to distinct immunological profiles between bovine hosts genetically susceptible and resistant. The influence of Bos indicus (Bi) and Bos taurus (Bt) maternal lineage ancestry of mitochondrial DNA in the profile of the immune response of Zebu cattle to ticks remains unknown. The present work evaluated the hematological parameters and the immune response profile in the peripheral blood of a Guzerat dairy herd, further categorized into two maternal lineage ancestry subgroups (Bi-mtDNA and Bt-mtDNA) after experimental infestation with larvae of R. microplus. Our data demonstrated that although hematological and erythrogram analysis showed a similar profile throughout, some cell populations present a distinct profile between the groups. Especially MON, CD335+ and CD8+ T-cells are predominant in Bi-mtDNA. Moreover, an overall picture of R. microplus infestation demonstrated that Bi-mtDNA presented a more efficient and earlier innate immune response. Bi-mtDNA showed a greater number of connections with R. microplus counts and also with the CD25+ activation marker of the immune response. Bi-mtDNA showed greater number of connections, with an important participation of the innate immune while Bt-mtDNA showed a delay in the immune response. Elucidating the mechanisms by which resistant animals prevent heavy tick infestation is a crucial step in the development of predictive biomarkers for tick resistance for use in selective breeding programs, and is also potentially useful for the development of anti-tick vaccines.

Baculoviral Expression of Presumptive OP-Resistance Mutations in BmAChE1 of Rhipicephalus (Boophilus) microplus (Ixodida: Ixodidae) and Biochemical Resistance to OP Inhibition.

The southern cattle tick, Rhipicephalus (Boophilus) microplus (Canestrini), transmits bovine babesiosis and anaplasmosis, and is endemic to Mexico, Latin and South America. Rhipicephalus (B.) microplus infestations within the United States are a continuing threat to U.S. cattle producers. An importation barrier between Texas and Mexico keeps the ticks from re-entering the United States. All cattle imported into the United States are dipped in an organophosphate (OP) acaricide and hand inspected for presence of ticks. Tick resistance has developed to most available acaricides, including coumaphos, the OP used in the cattle dip vats. OP-resistance can result from one or more mutations in the gene encoding the enzyme, acetylcholinesterase (AChE), resulting in production of an altered AChE resistant to OP inhibition. Previous research reported a large number of BmAChE1 mutations associated with OP resistance. We report baculovirus expression of recombinant tick BmAChE1 (rBmAChE) enzymes containing a single resistance-associated mutation, to assess their contribution to OP inhibition resistance. Surprisingly, of the naturally occurring BmAChE1 resistance-associated mutations, only D188G resulted in markedly reduced sensitivity to OP-inhibition suggesting that OP-insensitivity in BmAChE1 may result from the D188G mutation, or may possibly result from multiple mutations, each contributing a small decrease in OP sensitivity. Furthermore, an OP-insensitivity mutation (G119S) found in mosquitoes was expressed in rBmAChE1, resulting in 500-2000-fold decreased sensitivity to OP inhibition. Recombinant BmAChE1 with the G119S mutation demonstrated the lack of any structural prohibition to broad and high-level OP-insensitivity, suggesting potential increases in tick OP-resistance that would threaten the U.S. importation barrier to ticks.

Development of a new approach of pasture management to control Rhipicephalus microplus infestation.

Despite several decades of chemical control, the cattle tick Rhipicephalus microplus remains an important constraint for cattle farmers. The regular use of chemicals has led to the development of tick strains that are multi-resistant to acaricides. New methods of tick control are being developed and combined in integrated tick management programs. Grazing management is one part of these control methods. However, until now, grazing management consisted essentially of resting pastures for 4 to 5months. This amount of time is generally sufficient to allow for natural tick larvae mortality to occur, but this method often compromises the quality of forages, which is less than optimal at the time of arrival of the animals in the paddock. In this paper, we propose a new approach to pasture management-based tick control that optimizes forage production. It is based on tick development biological parameters, herd management and strategic treatments. This approach was tested for 2years on two farms raising tick-susceptible European breeds of cattle under tropical conditions. The number of chemical treatments decreased respectively by 82.9% and 70.9%. This cost-effective approach may prove useful in decreasing the number of acaricidal treatments on farms facing high tick loads.

Oral Vaccination With a Formulation Combining Rhipicephalus microplus Subolesin With Heat Inactivated Mycobacterium bovis Reduces Tick Infestations in Cattle.

Vaccines are an environmentally friendly alternative to acaracides for the control of tick infestations, to reduce the risk for tick-borne diseases affecting human and animal health worldwide, and to improve animal welfare and production. Subolesin (SUB, also known as 4D8) is the functional homolog of Akirin2 involved in the regulation of development and innate immune response, and a proven protective antigen for the control of ectoparasite infestations and pathogen infection. Oral vaccination combining protein antigens with immunostimulants has proven efficacy with increased host welfare and safety, but has not been used for the control of tick infestations. Here we describe the efficacy of oral vaccination with a formulation combining Rhipicephalus microplus SUB and heat inactivated Mycobacterium bovis (IV) on cattle tick infestations and fertility. The levels of IgG antibody titers against SUB and M. bovis P22, and the expression of selected immune response genes were determined and analyzed as possible correlates of protection. We demonstrated that oral immunization with the SUB+IV formulation resulted in 51% reduction in the number of female ticks and 30% reduction in fertility with an overall efficacy of 65% in the control of R. microplus infestations by considering the cumulative effect on reducing tick survival and fertility in cattle. The akr2, IL-1β, and C3 mRNA levels together with antibody levels against SUB correlated with vaccine efficacy. The effect of the oral immunization with SUB+IV in cattle on tick survival and fertility is essential to reduce tick infestations, and extended previous results on the effect of R. microplus SUB for the control of cattle tick infestations. These results support the development of oral vaccines formulations for the control of tick infestations and the incidence of tick-borne diseases.

Efficacy of winter-spring strategic control against Rhipicephalus (Boophilus) microplus infestations on cattle in an area with ecological conditions highly favourable for the tick in northeast Argentina.

This work was performed to test the efficacy of winter-spring control strategies against Rhipicephalus (Boophilus) microplus (Ixodida: Ixodidae) infestations on cattle in the area ecologically most favourable for the development of this tick in Argentina. Two control schemes using three and four annual applications of acaricides, respectively, were evaluated. Animals in Group 1 were treated with ivermectin 3.15% on day 0, fluazuron on day 34, and fipronil on day 85. Animals in Group 2 were treated with ivermectin 3.15% on day 0, fluazuron on day 34, flumethrin on day 85, and fipronil on day 114. Animals in Group 3 represented the control group. Both treatment schemes provided appropriate levels of efficacy against R. microplus and also prevented the occurrence of the major peak in the frequency of this tick in autumn. The two treatment schemes were similar in terms of efficacy and thus the addition of a fourth treatment does not seem to confer any further advantage. The results of this work indicate that these strategic control methods provide appropriate levels of control against R. microplus.

The dynamics of energy metabolism in the tick embryo.

The cattle tick Rhipicephalus (Boophilus) microplus is an ectoparasite capable of transmitting a large number of pathogens, causing considerable losses in the cattle industry, with substantial damage to livestock. Over the years, important stages of its life cycle, such as the embryo, have been largely ignored by researchers. Tick embryogenesis has been typically described as an energy-consuming process, sustaining cell proliferation, differentiation, and growth. During the embryonic stage of arthropods, there is mobilization of metabolites of maternal origin for the development of organs and tissues of the embryo. Glycogen resynthesis in late embryogenesis is considered as an effective indicator of embryonic integrity. In the cattle tick R.(B. (B.) microplus, glycogen resynthesis is sustained by protein degradation through the gluconeogenesis pathway at the end of the embryonic period. Despite recent advancements in research on tick energy metabolism at the molecular level, the dynamics of nutrient utilization during R. (B.) microplus embryogenesis is still poorly understood. The present review aims to describe the regulatory mechanisms of carbohydrate metabolism during maternal-zygotic transition and identify possible new targets for the development of novel drugs and other control measures against R. (B.) microplus infestations.

Anatomical distribution and population dynamics of Rhipicephalus (Boophilus) microplus in cattle in the municipality of Óleo, São Paulo

Rhipicephalus (Boophilus) microplus infestation is responsible for significant economic losses in cattle industry. The use of acaricides at strategic times and selection of resistant animals can minimize economic losses. This study aimed to determine the anatomical distribution and population dynamics of R. microplus in the municipality of Óleo, state of São Paulo. This study was conducted using eight male cattle, crossbred zebu-taurine (in accordance with the regional herd). Female R. (B.) microplus, greater than 4 mm, present in the right side of the animals were counted between April 2012 and March 2013. The highest tick number per animal was noted in October (128.25), and the lowest in April (35.5), with three infestation peaks during the study, in May, October, and December, which were influenced by the minimum and maximum mean temperature and rainfall. Among the cranio-caudal regions (anterior, median, and posterior), the posterior had the highest mean tick number, and the anterior had the lowest. The minimum temperature, rainfall, and solar radiation interfered with infestation of dorsoventral regions: in cold months, the lowest infestations were found in the mixed area (ear and tail); in the warm months, in the high region (head, neck, upper ribs, flank, and rump). The lower region (dewlap, cranial limb, lower ribs, abdomen, prepuce, scrotum, and lower limb) always had the highest mean tick number. The tick numbers of the caudal limb and crotch showed strong correlation with the total tick number (+0.88 and +0.70, respectively). These data, grouped according to median, posterior, and crotch regions, showed a stronger correlation with the total tick number (+0.96, +0.90, and +0.70, respectively).

Molecular characterization of the recombinant protein RmLTI-BmCG-LTB: Protective immunity against Rhipicephalus (Boophilus) microplus.

The bovine tick Rhipicephalus (Boophilus) microplus is found in several tropical and subtropical regions of the world. This parasite transmits pathogens that cause disease, such as babesiosis (Babesia bovis and B. bigemina) and anaplasmosis (Anaplasma marginale). Tick infestations cause enormous livestock losses, and controlling tick infestations and the transmission of tick-borne diseases remains a challenge for the livestock industry. Because the currently available commercial vaccines offer only partial protection against R. (B.) microplus, there is a need for more efficient vaccines. Several recombinant antigens have been evaluated using different immunization strategies, and they show great promise. This work describes the construction and immunological characterization of a multi-antigen chimera composed of two R. (B.) microplus antigens (RmLTI and BmCG) and one Escherichia coli antigen (B subunit, LTB). The immunogenic regions of each antigen were selected and combined to encode a single polypeptide. The gene was cloned and expressed in E. coli. For all of the experiments, two groups (treated and control) of four Angus heifers (3–6 months old) were used. The inoculation was performed via intramuscular injection with 200 μg of purified recombinant chimeric protein and adjuvated. The chimeric protein was recognized by specific antibodies against each subunit and by sera from cattle inoculated with the chimera. Immunization of RmLTI-BmCG-LTB cattle reduced the number of adult female ticks by 6.29% and vaccination of cattle with the chimeric antigen provided 55.6% efficacy against R. (B.) microplus infestation. The results of this study indicate that the novel chimeric protein is a potential candidate for the future development of a more effective vaccine against R. (B.) microplus.

Acaricidal potency of polyherbal spray against Rhipicephalus microplus and Hyalomma anatolicum infestation in cattle.

Acaricidal potential of polyherbal spray (Andropogon citrates, Cymbopogon citratus, Ocimum sanctum, Pinus longifoia, Calotropis procera, Datura stramonium, Aegle marmelos, Ricinus communis, Azadirachta indica, Allium sativum, Carica papaya, Annona squamosa and Pongamia glabra) was assessed against tick infestation in cattle on the basis of measurement of tick count, complete blood count and plasma glucose, total protein, albumin and globulin before treatment and 21days after treatment. Single application of polyherbal spray over body of 20 randomly selected tick infested cattle revealed significant reduction in mean tick count starting from 3days post treatment till 21days post treatment. Highly significant (P<0.01) increase in total erythrocyte count and packed cell volume was observed in treated cows 21days after application of spray compared to pre-treatment values indicating the reduction in blood loss due to heavy tick infestation before treatment. Plasma biochemical parameters revealed no significant changes in pre-treatment and post treatment values. The results of present study imply the clinical and haematological improvement in tick infested cattle treated with polyherbal spray and it could be potential product for use in livestock as acaricide.

Functional characterization of candidate antigens of Hyalomma anatolicum and evaluation of its cross-protective efficacy against Rhipicephalus microplus.

Hyalomma anatolicum and Rhipicephalus microplus seriously affect dairy animals and immunization of host is considered as a sustainable option for the management of the tick species. Identification and validation of protective molecules are the major challenges in developing a cross-protective vaccine. The subolesin (SUB), calreticulin (CRT) and cathepsin L-like cysteine proteinase (CathL) genes of H. anatolicum were cloned, sequenced and analysed for sequence homology. Both Ha-SUB and Ha-CRT genes showed very high level of homogeneity within the species (97.6-99.4% and 98.2-99.7%) and among the tick species (77.3-99.3% and 85.1-99.7%) while for Ha-CathL the homogeneity was lower among ticks (57.5-89.5%). Besides tick species, both Ha-SUB and Ha- CRT genes showed high level of homogeneity with dipterans (47.2-53.4% and 72.0-74.4%) and nematodes (64.0% by CRT). The level of expression of the conserved genes in different stages of the tick species was studied. The differences in fold change of expression (FCE) of the targeted genes in life stages of tick were not statistically significant except Ha-SUB in eggs and in frustrated females, Ha-CRT in fed male and Ha-CathL in unfed and frustrated females where highest FCE was recorded. The functional properties of the genes were studied by RNAi technology and a significant level of gene suppression (p<0.05) resulted in very low percentage of engorgement of treated ticks viz., 3.7%, 11.1% and 30.0% in Ha-SUB, Ha-CRT and Ha-CathL respectively, in comparison to control was recorded. The recombinant proteins rHa-SUB, rHa-CRT and rHa-CathL encoded by the genes were expressed in prokaryotic expression system. They were evaluated for cross-protective efficacy and found to be respectively, 65.4%, 41.3% and 30.2% protective against H. anatolicum and 54.0%, 37.6% and 22.2%, against R. microplus infestations.

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

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 methods to control R. microplus infestation on cattle.

Off-host development and survival of Rhipicephalus (Boophilus) microplus in the Brazilian semiarid.

Rhipicephalus (Boophilus) microplus is the most economically important tick of the world, including Brazil. Whereas the epidemiology of R. microplus is well known in most of the Brazilian land, virtually nothing is known from the Brazilian semiarid region, the Caatinga biome. Considering the relevance of R. microplus for the cattle industry within the Caatinga, this study aimed to evaluate for the first time the off-host development of this tick species under natural conditions of the Caatinga. During 2011-2015, engorged females of R. microplus were exposed to field conditions of native Caatinga, where female oviposition, egg incubation and hatching, and larval survival were quantified. In parallel, counterpart ticks (control group) were evaluated under optimal conditions in the laboratory. During the study, nearly 100% of the engorged females of the control group successfully oviposited fertile egg masses, from which most of the eggs yielded larvae (mean % hatching usually>90%). Under field conditions, while almost 100% of the engorged females oviposited, in most of the times no larvae hatched from these eggs. Furthermore, when larvae hatched, mean % hatching was most of the times <50%. Soil temperatures did not oscillate drastically through the study, with mean temperature between 30 and 35°C in most of the time. In contrast, rainfall was irregular throughout the study. Correlation analyses indicated that either the number of egg masses that yielded larvae, or the mean % hatching of the egg masses were positively associated with higher rainfall months. Our results showed that in most of the time, field conditions were unfavorable for R. microplus, indicating that cattle would have a low exposure to R. microplus infestations when raised in Caatinga fields like those of the present study.

Levels od natural resistance to Boophilus microplus( Acari: Ixodidae) in Carrora breed bulls.

Boophilus microplus infestation is one of the most serious limitations to cattle industry in tropical regions, even though bovines show natural resistance to ticks. This resistance was evaluated in Cross-bred Carora Bulls (CCB) a tropicalized dairy breed from Venezuela. Seven CCB were experimentally infested with B. microplus larvae, "Mozo" strain, they were considered tick-naive because they had never been infested with ticks. The mean inoculum size applied on each bull was 6,477 larvae. After life cycle was completed adult female body weight (BW), egg mass weight (EW), egg hatching rate (%EH), and reproductive index (RI) were recorded. Results revealed a high variability in the levels of resistance to B. microplus. Thus, one animal showed greater resistance (Dunnett, p < 0.05) for the analyzed parameters in contrast with three non-resistant bulls. The others had moderate resistance. The trait "resistance" should be included together with other traits often used in genetic selection of cattle.

Immunoprotective potential of a Rhipicephalus (Boophilus) microplus metalloprotease

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.