Brazil's battle against Rhipicephalus (Boophilus) microplus ticks: current strategies and future directions.

Ticks are parasitic arthropods that cause significant economic losses to livestock production worldwide. Although Rhipicephalus (Boophilus) microplus, the cattle tick, occurs throughout the Brazilian territory, there is no official program to control this tick, which is the vector of tick fever pathogens. We address the situation of R. (B.) microplus resistance to synthetic acaricides in Brazil, including cattle tick management; the status of tick resistance per Brazilian state; the history of resistance occurrence of different acaricides; multiple resistance occurrence; and the main strategies for integrated tick management. Tick control in Brazil is characterized by management errors. Local laboratories affiliated with federal and state research institutions and universities employ the Adult Immersion Test as a primary diagnostic method to assess acaricide resistance to topically applied drugs. Only three states (Acre, Amapá, and Amazonas) have no reports on resistant populations. Misinformation on tick control strategies, misuse of available products for tick control, no adoption of Integrated Parasite Management (IPM) practices, low technical support to producers, and the high-speed emergence of acaricide-resistant tick populations are the main problems. We also propose a list of needs and priorities for cattle tick control regarding communication, research, and policies.

Can herd seroprevalence be used as an indicator of enzootic stability for bovine anaplasmosis? Insights from a case-control field study in Brazil.

Bovine anaplasmosis presents a significant challenge to livestock production in tropical, subtropical, and temperate regions. For many years, the concept of enzootic stability/instability (initially established for babesiosis) and herd seroprevalence as an indicator of outbreak risks have been applied to anaplasmosis. However, this model has never been definitively validated for Anaplasma marginale. The objective of this study was to examine the relationship between herd immunity (seroprevalence) and the occurrence of anaplasmosis outbreaks in Southern Brazil. A case-control study was conducted, categorizing farms into two groups: cases (farms with a history of clinical anaplasmosis) and controls (those without anaplasmosis). Thirteen farms were identified as "cases", while 23 were identified as "controls". A substantial difference in seroprevalence distribution between the two groups was observed. The majority of "control" farms exhibited over 75% of animals with antibodies to A. marginale in both calves and heifers, whereas the majority of "case" farms had a seropositive cattle percentage below 75%. Additionally, twelve months after cattle serology tests, we conducted a prospective follow-up survey to identify any clinical cases of anaplasmosis. Statistical associations (P < 0.05) were found between both retrospective and prospective anaplasmosis outbreaks and the hypothetical threshold of herd seroprevalence (75%). We hypothesize that herd seroprevalence may be an indicator of the risk of occurrence of clinical anaplasmosis. It appears that the epidemiology of cattle anaplasmosis, at least in our conditions, aligns with the well-known model of enzootic stability/instability originally applied to bovine babesiosis.

Does parity order or cow antibody level influence the humoral immunity of calves for cattle anaplasmosis?

Anaplasmosis is a widely distributed vector-borne disease of cattle caused by the bacteria Anaplasma marginale, which may lead to severe losses in beef and dairy cattle production. Anecdotal information among farmers suggested that some cows may deliver calves more resilient to anaplasmosis. To investigate this, we tested two hypotheses: (i) whether the parity order or (ii) cow antibody levels may influence the humoral immunity of pre-weaning calves against cattle anaplasmosis. For this study, we sampled 170 cattle (Bos taurus taurus, Angus breed) on a farm in Southern Brazil, comprising 85 cows (50 multiparous and 35 primiparous) and their 85 calves (50 days old). Antibodies against A. marginale were investigated using iELISA. Eighty percent of the animals were considered seropositive (100% of the cows and 60% of the calves). There was no significant difference in seroprevalence between calves from primiparous and multiparous cows. However, calves from multiparous cows exhibited higher antibody levels. To address the second question, we classified cows based on their antibody levels to A. marginale (high or low). Calves from cows with high antibody levels also showed elevated antibody levels against A. marginale. Furthermore, calves from cows with high antibody levels had approximately four times greater odds of being seropositive for A. marginale at 50 days old than those born to mothers with low antibody levels. In conclusion, the calf's antibody level against A. marginale appears to be related to the cow's parity order and the mother's antibody level.

Phenotypic and genotypic characterization of acaricide resistance in Rhipicephalus microplus field isolates from South Africa and Brazil.

Rhipicephalus (Boophilus) microplus is one of the most successful ticks infesting cattle around the world. This highly-invasive species transmits cattle parasites that cause cattle fever leading to a high socio-economic burden. Tick eradication programs have often failed, due to the development of acaricide resistance. Here we characterize acaricide resistance in a large number of tick isolates from regions in South Africa (KwaZulu Natal, Mpumalanga, Western & Eastern Cape provinces) and two Brazilian regions. By means of Larval Packet Tests (LPT's) acaricide resistance was evaluated against five commonly used acaricides (chlorfenvinphos, fipronil, deltamethrin, amitraz, and ivermectin). Furthermore, the coding region containing the knock down resistance (kdr) mutation, known to result in pyrethroid resistance, was sequenced. Resistance to at least one acaricide class was reported in each of the five regions, and a high proportion of tick isolates exhibited multi-resistance to at least two acaricide classes (range: 22.2-80.0%). Furthermore, resistance ratios (RR) showed high spatial variation (intercontinental, as well as regional) but low regional spatial autocorrelation. Previous and current acaricide use correlated with current RR, and several combinations of acaricide RR were positively correlated. Moreover, fipronil resistance tended to be higher in farms with more intense acaricide use. The kdr-mutations provided the ticks a fitness advantage under the selection pressure of synthetic pyrethroids based on population (kdr-allele frequency) and individual level data (genotypes). The data show the threat of acaricide (multi-)resistance is high in Brazil and South Africa, but acaricide specific levels need to be assessed locally. For this purpose, gathering complementary molecular information on mutations that underlie resistance can reduce costs and expedite necessary actions. In an era of human-caused habitat alterations, implementing molecular data-driven programs becomes essential in overcoming tick-induced socio-economic losses.

Molecular Survey of Haemosporidian Parasites in Procellariiformes Sampled in Southern Brazil, 2013-22.

The order Procellariiformes includes several species of seabirds that perform long-distance migrations crossing all the oceans. These movements may contribute to the dispersal and exchange of hemoparasites, such as haemosporidians. There is a lack of studies regarding the order Haemosporida in Procellariiformes, and, to date, only the genus Plasmodium has been reported. This survey investigated the occurrence of the three genera of haemosporidians, Plasmodium, Haemoproteus, and Leucocytozoon, in samples collected between 2013 and 2022 from 95 individuals of 14 species of Procellariiformes from southern Brazil, including live animals in rehabilitation centers, individuals caught as incidental bycatch, and carcasses found along the coast. A total of 171 samples of blood and fragments of liver and spleen were analyzed, with extracted DNA being subjected to a nested PCR followed by phylogeny analysis. All animals were negative for Plasmodium spp. and Leucocytozoon spp., but one Black-browed Albatross (Thalassarche melanophris) and one Manx Shearwater (Puffinus puffinus) specimen were positive for Haemoproteus spp. The sequences obtained from positive seabirds did not show 100% similarity with other known lineages available in the MalAvi database and thus were probably novel lineages. However, one sequence clustered together with Haemoproteus noctuae, a parasite from Strigiformes, while the other was grouped with Haemoproteus columbae, which is classically related to Columbiformes. These results suggest that both positive animals may have become infected when beached or in rehabilitation centers by a spillover of vectors from local birds. This highlights the importance of surveillance of the health of Procellariiformes regarding the possibility of dissemination of new pathogens in different bird populations.

Selective versus strategic control against Rhipicephalus microplus in cattle: A comparative analysis of efficacy, animal health, productivity, cost, and resistance management.

This study compared selective control versus strategic control against Rhipicephalus (Boophilus) microplus, under conditions of a real dairy farm. Strategic control requires that all animals be treated with acaricide at regular pre-determined intervals. Selective control only requires treatment of infested animals and only when they are at or above a pre-determined threshold. Tick counts on animals and in pasture were performed and the susceptibility of tick populations to the different treatment methods was evaluated at the beginning and end of the study using the Larval Packet Test, Larval Immersion Test, and Adult Immersion Test. Over the four years of the study strategic control was more advantageous than the selective control as the group experienced lower tick burden on animals and in pasture, absence of skin lesions and myiases, lower operating costs, treatments concentrated in fewer months of the year and lower resistance pressure.

A novel variant of Babesia sp. (Piroplasmida) as a hemoparasite in procellariiform seabirds.

Procellariiformes includes pelagic seabirds that only use land for breeding; and also, these sites mostly occur in insular habitats. These peculiar habits make the investigation of hemoparasites a challenging issue. Thus, the data on the blood parasites of Procellariiformes are still scarce. In the order Piroplasmida, 16 species of Babesia have been described in terrestrial birds and seabirds. However, there is no register for Babesia spp. in procellariiform seabirds. Hence, the objective of this survey was to investigate the occurrence of Babesia spp. in these seabirds. A total of 220 tissue samples from 18 different seabird species were analyzed; the samples comprised blood and fragments of liver and spleen. The samples were obtained from live rescued animals and carcasses found along the southern coast of Brazil. Polymerase chain reaction (PCR) was conducted, followed by phylogenetic analysis. Only one blood sample yielded a positive result, from an adult female Thalassarche chlororhynchos (Atlantic yellow-nosed albatross). The sequence obtained showed the highest identity with sequences of Babesia spp. of birds from the South Pacific, and the isolate was named Babesia sp. strain Albatross. In the phylogenetic analysis, the sequence was grouped within the Babesia sensu stricto group, and further still into a subgroup including Babesia spp. of the Kiwiensis clade (parasites from birds). The phylogenetic analysis also showed that Babesia sp. strain Albatross clustered apart from the Peircei group, a clade that includes Babesia spp. from seabirds. As far as it is known, this is the first report of Babesia sp. in procellariiform seabirds. Babesia sp. strain Albatross may constitute a novel variant of tick-borne piroplasmids associated with the Procellariiformes order.

Automatic method based on deep learning to identify and account Rhipicephalus microplus larval hatching.

Reports of Rhipicephalus microplus resistant populations worldwide have increased extensively, making it difficult to control this ectoparasite. The adult immersion test, commonly used to screen for acaricide resistance, produces the results only after 40 days of the tick collection because it needs the eggs to be laid and larvae to hatch. The present study aims to develop an automatic method, based on deep learning, to predict the hatching of R. microplus larva based on egg morphology. Initially, the time course of embryonic development of tick eggs was performed to discriminate between viable and non-viable eggs. Secondly, using artificial intelligence deep learning techniques, a method was developed to classify and count the eggs. The larval hatching rate of three populations of R. microplus was evaluated for the software validation process. Groups of three and six images of eggs with 12 days of embryonic development were submitted to the software to predict the larval hatching percent automatically. The results obtained by the software were compared with the prediction results of the hatching percentage performed manually by the specialist and with the results of the hatching percentage of larvae obtained in the biological assay. The group with three images of each population submitted to the software for automatic prediction of the larval hatching percent presented mean values of 96.35% ± 3.33 (Piracanjuba population), 95.98% ± 3.5 (Desterro population) and 0.0% ± 0.0 (Barbalha population). For groups with six images, the values were 94.41% ± 3.84 (Piracanjuba population), 95.93% ± 2.36 (Desterro population) and 0.0% ± 0.0 (Barbalha population). Biological assays showed the following hatching percentage values: 98% ± 1.73 (Piracanjuba population); 96% ± 2.1 (Desterro population); and 0.14% ± 0.25 (Barbalha population). There was no statistical difference between the evaluated methods. The automatic method for predicting the hatching percentage of R. microplus larvae was validated and proved to be effective, with considerable reduction in time to obtain results.

Comparison of Seroprevalence and Identification of Risk Factors for Theileria equi in Horses From Vector-Free and Infested Areas in Southern Brazil.

The apicomplexan hemoprotozoan Theileria equi is a tick-borne pathogen that causes disease in equids, and together with Babesia caballi, causes equine piroplasmosis (EP). Many ticks are associated with EP around the world, and in South America three species may be related: Dermacentor nitens, Amblyomma sculptum, and Rhipicephalus microplus, as they are commonly found in horses. Among the species cited above, only R. microplus is found in Rio Grande do Sul state, Southern Brazil. In addition, this state has the only area legally free of R. microplus in Brazil. This study aimed to compare the seroprevalence for T. equi on farms located in a tick-free area (23 farms, 215 horses) and a tick-infested (25 farms, 141 horses) adjacent areas, as well as to identify potential risk factors for exposure to T. equi. Blood was collected from 356 horses from both areas, and later an indirect enzyme-linked immunosorbent assay was performed to detect anti-T. equi antibodies. Besides the blood collection, questionnaires and interviews were conducted in an attempt to identify potential risk factors. The R. microplus-free zone had 6% seropositive horses against 70% in the tick-infested zone. Previous observation of ticks on horses of the herd increased four times the odds of a horse to be seropositive for T. equi, and by three times if ticks were observed on cattle which share paddocks with horses on the farm. The results showed a large difference in T. equi seroprevalence between tick-infested and free areas, and also emphasized the relevance of R. microplus in the epidemiology of T. equi. The study also reveals the potential of the establishment of a T. equi-free zone for horse breeding in Southern Brazil.

Putative target sites in synganglion for novel ixodid tick control strategies.

Acaricide resistance is a global problem that has impacts worldwide. Tick populations with broad resistance to all commercially available acaricides have been reported. Since resistance selection in ticks and their role in pathogen transmission to animals and humans result in important economic and public health burden, it is essential to develop new strategies for their control (i.e., novel chemical compounds, vaccines, biological control). The synganglion is the tick central nervous system and it is responsible for synthesizing and releasing signaling molecules with different physiological functions. Synganglion proteins are the targets of the majority of available acaricides. In this review we provide an overview of the mode-of-action and resistance mechanisms against neurotoxic acaricides in ticks, as well as putative target sites in synganglion, as a supporting tool to identify new target proteins and to develop new strategies for tick control.

Effect of cyclosporin A on the toxicity of ivermectin, eprinomectin and moxidectin in populations of Rhipicephalus microplus.

Rhipicephalus microplus is mainly controlled by acaricides. However, reports of resistance to acaricides including macrocyclic lactones (MLs) have become frequent worldwide. Involvement of ABC transporters (ABCts) in populations resistant to ivermectin has been demonstrated. Thus, the aim of this study was to evaluate the efficacy of ivermectin, eprinomectin and moxidectin with and without use of synergistic cyclosporin A (CsA) in resistant populations of R. microplus using larval immersion tests (LITs). Engorged females were collected from four farms in the semiarid region of northeastern Brazil that had histories of continuous use of ivermectin. Questionnaires were applied to collect information about management aimed at controlling ticks on these farms. Resistance to MLs was observed on all of the farms. There was statistically significant synergism (p < 0.05) between CsA and ivermectin in all populations; between CsA and eprinomectin in only one population; and between CsA and moxidectin in two populations. It was concluded that, despite the involvement of ABCts in the mechanisms of resistance to ivermectin, metabolic detoxification does not seem to be the mechanism predominantly involved in resistance to eprinomectin and moxidectin in the populations of R. microplus evaluated.

Interaction between anti-tick vaccine and a macrocyclic lactone improves acaricidal efficacy against Rhipicephalus (Boophilus) microplus (Canestrini) (Acari: Ixodidae) in experimentally infested cattle.

The southern cattle fever tick (SCFT) Rhipicephalus (Boophilus) microplus, is considered the most important ectoparasite of livestock in the world because of high financial losses associated with direct feeding and transmission of the hemoparasites Babesia bovis, B. bigemina, and Anaplasma marginale. Unfortunately, SCFT in many parts of the world have evolved resistance to all market-available pesticides thus driving development of new control technologies. Vaccination against ticks using the tick gut protein Bm86 has been shown to be effective against acaricide-resistant ticks. This technique has been successfully implemented in Puerto Rico for the control of acaricide-resistant R. microplus on dairy and beef cattle. Observations from Puerto Rico indicate a potentially positive interaction between anti-tick vaccination when used in conjunction with systemic acaricide treatment. In this project, controlled animal studies were completed directly comparing efficacy of anti-tick vaccination with and without systemic acaricide. Results show that the Bm86 anti-tick vaccine in combination with the macrocyclic lactone, Moxidectin, expressed a synergistic interaction, providing greater and longer efficacy than either treatment alone.

Cross-resistance between macrocyclic lactones in populations of Rhipicephalus microplus in Brazil's semiarid region.

Rhipicephalus microplus, also known as the cattle tick, is the parasite with the greatest impact on cattle in Brazil. The most common method for controlling this tick is the application of synthetic chemical acaricides, especially ivermectin, which belongs to the group of macrocyclic lactones (MLs). However, because ivermectin is widely used, there is concern about the development of cross-resistance within this chemical class. Thus, engorged females were collected from farms with a history of resistance to ivermectin, which was the only one among the MLs that was used as an endectocide drug. Using larval immersion tests (LIT), bioassays were performed with ivermectin, moxidectin and eprinomectin on populations of R. microplus from the semiarid region of the states of Paraíba and Ceará, Brazil. Epidemiological questionnaires were applied to collect information about tick control management. All the evaluated populations showed cross-resistance between ivermectin and moxidectin, but only one population showed cross-resistance between ivermectin and eprinomectin. Weekly or monthly administration of injectable 1% ivermectin on farms was reported. It was concluded that the frequent use of ivermectin may lead to the development of cross-resistance to moxidectin. For eprinomectin, despite the structural similarity, cross-resistance was not observed in three tick populations.

Development and validation of software that quantifies the larval mortality of Rhipicephalus (Boophilus) microplus cattle tick.

Rhipicephalus (Boophilus) microplus is controlled almost exclusively using synthetic acaricides, and reports of resistant populations have been described worldwide. Several time-consuming and laborious toxicological in vitro tests have been used to diagnose acaricidal resistance, especially those that require differential counting of live and dead larvae. Larval mortality is currently done manually and subjectively, which can limit the performance of a large number of tests and comparing results between different laboratories. The present study aimed to develop and validate a new automatic counting method to evaluate tick larval mortality. A software for differentiation of live and dead larvae was developed using different steps: obtaining videos; image segmentation using the firefly algorithm; detection of larvae with the fast radial symmetry transform technique (FRST); and tracking of the larvae at a given time. Larval immersion tests with ivermectin, cypermethrin, and fipronil were performed to validate the developed software. The larval mortality evaluation was performed by (1) recording for 60 s for each package and (2) manual counts of the same sample using three different analysts, each responsible for counting one replicate of each test. All videos obtained were copied and cut at 60, 40, and 20 s for later analysis in the counting software. The median lethal doses (LD50) of the different compounds in each test were calculated for each method (automatic and manual) for different video times. There was no statistical difference in LD50 between manual and automatic count techniques for ivermectin and fipronil. The LD50 of cypermethrin calculated with manual evaluation was up to 2.2 times lower than that of automatic evaluation. The acquisition time of the videos was 2.9-4.4 times faster than the manual evaluation. The average processing time for each video was 5.73 min, regardless of their duration. Thus, the method developed for automatic counting of tick larvae was validated, and although it still has points to be optimized, it can be considered a viable alternative for determining the percentage of tick larvae mortality and could be applied to toxicological in vitro tests with acaricides, assisting in the diagnosis of resistant tick populations and studies of novel acaricide development.

Mechanisms of amitraz resistance in a Rhipicephalus microplus strain from southern Brazil.

Amitraz is one of the most used acaricides for the control of ticks of domestic animals, however, extensive use of this active ingredient has favored the development of resistant populations of Rhipicephalus microplus worldwide. The possible mechanisms of metabolic and/or target-site alterations mechanisms of amitraz resistance were investigated in a Brazilian field population of R. microplus (São Gabriel strain). Bioassays with the synergists piperonylbutoxide, triphenylphosphate and diethyl-maleate were used to evaluate the metabolic mechanisms involved. Target-site insensitivity was investigated by amplification and sequencing of a fragment of the octopamine/tyramine (OCT/TYR) receptor gene. Piperonylbutoxide synergism (synergism ratio = 2.8) indicated the participation of the P450 pathway in the detoxification of amitraz. Previously reported single nucleotide polymorphisms that confer amino acid changes in the OCT/TYR receptor, threonine to proline (T8P) and leucine to serine (L22S), were found in the amitraz-resistant strain but not in the susceptible reference strain. The results suggest that amitraz resistance in the studied strain is multi-factorial and may result from cytochrome P450 detoxification and mutations in octopamine receptors.

Efficacy of a water-based botanical acaricide formulation applied in portable spray box against the southern cattle tick, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae), infesting cattle.

Ectoparasitism result in annual losses to livestock producers estimated to reach billions of dollars in tropical and subtropical parts of the world where the southern cattle tick, Rhipicephalus (Boophilus) microplus, is established because this invasive pest is also a vector of pathogens causing bovine babesiosis and anaplasmosis. Reintroduction of R. microplus could be economically devastating for the cattle industry in the United States. Novel technologies are necessary to manage populations of R. microplus that are resistant to multiple classes of synthetic chemicals widely used as acaricides to control ticks infesting livestock, and to mitigate the environmental impact of these synthetic acaricides. Bioassays with a water-based formulation of a commercially available botanical acaricide (Essentria® IC3) at 6.25 % against a laboratory strain of R. microplus was 100 % lethal against unfed larvae, and 94 % mortality was recorded against engorged female ticks. These results prompted documentation of efficacy on infested cattle using an acaricide delivery system mimicking field conditions, which is required to consider the use of a product by integrated tick management programs. Twelve tick naïve cattle were artificially infested with unfed larvae on days -19, -12 and -5. On day -2, ticks from the left-side of the body of each animal were manually counted. Depending on pre-treatment tick counts the animals were assigned into three separate treatment groups: i) water (untreated control); ii) coumaphos 0.3 % in water (positive control); and iii) botanical acaricide 6.25 % in water. Five days after the last infestation, cattle were doused in a spray box and moved to individual stalls to collect tick data for calculation of treatment efficacy. The percentage control, based on the reproduction of surviving engorged female ticks after treatment with the 6.25 % Essentria in water-based spray was 70 % against R. microplus. Coumaphos treatment had a mean percent control of 100 %. Whereas this application of Essentria® IC3 cannot be recommended as a stand-alone method to eradicate R. microplus, the 6.25 % botanical acaricide water-based spray could be part of integrated tick management to control populations of R. microplus resistant to organophosphates like coumaphos and other classes of commercially available synthetic acaricides.

Multiplex allele-specific polymerase chain reaction to detect kdr and rdl mutations in the cattle tick Rhipicephalus microplus (Acari: Ixodidae).

The resistance of Rhipicephalus microplus to pyrethroids is widely dispersed worldwide and has been associated with several nucleotide substitutions in its target site, the para-sodium ion channel (Na-channel) gene. The resistance of the tick to fipronil has been increasing in South America, and mutations in the GABA-gated chloride channel (GABA-Cl) have been described in fipronil-resistant tick strains. We developed a multiplex allele-specific PCR (mAS-PCR) to screen for single-nucleotide polymorphisms (SNPs) associated with the resistance to pyrethroids (knockdown resistance or kdr) and fipronil (resistance to dieldrin or rdl) in susceptible tick populations from Uruguay (n = 11) and the Rio Grande do Sul state in Southern Brazil (n = 15). Toxicological in vitro assays with larvae and adults were used to confirm the resistance to cypermethrin, flumethrin, and fipronil. Three SNPs in the Na-channel gene were investigated (C190A, G215 T, and T2134A), and the mAS-PCR included the detection of an SNP (G858 T) coding a non-synonymous mutation in the GABA-Cl gene. C190A was present in all pyrethroid-resistant populations from Uruguay and Brazil, most frequently homozygous. The SNPs G215 T and T2134A were not found. Of the seventeen fipronil-resistant populations, fourteen presented at least one mutant GABA-Cl gene allele, more frequently in heterozygosis. Other mechanisms apart from target site insensitivity may be involved in fipronil resistance since in some resistant populations, the SNP G858 T was not detected. Sixteen (61,5%) of the populations presented individuals with simultaneous mutations in the Na-channel and GABA-Cl genes. This could be a significant problem for the future control of R. microplus. This study shows the wide dispersion of a pyrethroid resistance-associated SNP in high frequency in the region. Fipronil resistance mutations are also dispersed across the region and increasing.

Susceptibility of field populations of Haematobia irritans to fipronil in Uruguay

ABSTRACT: Fipronil was registered in Uruguay in 1997, and, since then, it has been used for the control of Haematobia irritans irritans and Rhipicephalus microplus. The susceptibility of H. irritants to this drug has not been evaluated. Therefore, the goal of the present study was to evaluate the resistance of H. irritans to fipronil. Additionally, a survey was carried out with the farmers to evaluate the use of fipronil for H. irritans control in the ranches where the flies came from. For the bioassays, 31 field populations of H. irritans were exposed to 10 concentrations of fipronil (3.2-16.0g.cm2), and their LC50 values were calculated using probit analysis. A bioassay was performed with horn flies from the susceptible colony maintained at the USDA-ARS Knipling-Bushland U.S. Livestock Insects Research Laboratory for comparison and calculation of resistance ratios (RRs). All 31 field populations surveyed in the study were susceptible to fipronil, with resistance ratios ranging from 0.5 to 2.2. Four populations with RRs >1 did not differ significantly from the susceptible strain. A single population showed an RR >2.2. Overall, the survey shows that fipronil was mostly used for R. microplus control, and in only three ranches, which were free of R. microplus, was fipronil used for horn fly control. Seventeen farmers did not use fipronil at all in the last three years. It is concluded that, in Uruguay, field populations of horn flies remain susceptible to fipronil.

RESUMO: O fipronil foi registrado no Uruguai em 1997 e, desde então, tem sido utilizado no controle de Haematobia irritans irritans e Rhipicephalus microplus. O objetivo do presente estudo foi avaliar a susceptibilidade de populações de campo de H. irritans ao fipronil. Além disso, foi realizada uma pesquisa para avaliar a utilização de fipronil e as práticas de controle de H. irritans nas fazendas de onde provinham as moscas. Para os bioensaios, 31 populações de campo de H. irritans foram expostas a 10 concentrações de fipronil (3,2-16,0g.cm2), e seus valores de CL50 foram calculados usando análise probit. Um bioensaio foi realizado com H. irritans da colônia suscetível mantida no USDA-ARS Knipling-Bushland U.S. Livestock Insects Research Laboratory para comparação e cálculo das razões de resistência (RRs). Todas as 31 populações de campo pesquisadas no estudo eram suscetíveis ao fipronil, com taxas de resistência variando de 0,5 à 2,2. Quatro populações com Rrs >1 não diferiram significativamente da cepa suscetível. Uma única população apresentou RR >2,2. No geral, o fipronil tinha sido usado principalmente para o controle de R. microplus, e em apenas três fazendas, que estavam livres de R. microplus, o fipronil era utilizado para o controle da H. irritans. Em 17 fazendas não tinha sido utilizado fipronil nos últimos três anos. Conclui-se que no Uruguai as populações de H. irritans no campo permanecem suscetíveis ao fipronil.

Susceptibility of field populations of Haematobia irritans to fipronil in Uruguay / Suscetibilidade de populações de campo de Haematobia irritans ao fipronil no Uruguai

Fipronil was registered in Uruguay in 1997, and, since then, it has been used for the control of Haematobia irritans irritans and Rhipicephalus microplus. The susceptibility of H. irritants to this drug has not been evaluated. Therefore, the goal of the present study was to evaluate the resistance of H. irritans to fipronil. Additionally, a survey was carried out with the farmers to evaluate the use of fipronil for H. irritans control in the ranches where the flies came from. For the bioassays, 31 field populations of H. irritans were exposed to 10 concentrations of fipronil (3.2-16.0μg.cm2), and their LC50 values were calculated using probit analysis. A bioassay was performed with horn flies from the susceptible colony maintained at the USDA-ARS Knipling-Bushland U.S. Livestock Insects Research Laboratory for comparison and calculation of resistance ratios (RRs). All 31 field populations surveyed in the study were susceptible to fipronil, with resistance ratios ranging from <0.5 to 2.2. Four populations with RRs >1 did not differ significantly from the susceptible strain. A single population showed an RR >2.2. Overall, the survey shows that fipronil was mostly used for R. microplus control, and in only three ranches, which were free of R. microplus, was fipronil used for horn fly control. Seventeen farmers did not use fipronil at all in the last three years. It is concluded that, in Uruguay, field populations of horn flies remain susceptible to fipronil.(AU)

O fipronil foi registrado no Uruguai em 1997 e, desde então, tem sido utilizado no controle de Haematobia irritans irritans e Rhipicephalus microplus. O objetivo do presente estudo foi avaliar a susceptibilidade de populações de campo de H. irritans ao fipronil. Além disso, foi realizada uma pesquisa para avaliar a utilização de fipronil e as práticas de controle de H. irritans nas fazendas de onde provinham as moscas. Para os bioensaios, 31 populações de campo de H. irritans foram expostas a 10 concentrações de fipronil (3,2-16,0μg.cm2), e seus valores de CL50 foram calculados usando análise probit. Um bioensaio foi realizado com H. irritans da colônia suscetível mantida no USDA-ARS Knipling-Bushland U.S. Livestock Insects Research Laboratory para comparação e cálculo das razões de resistência (RRs). Todas as 31 populações de campo pesquisadas no estudo eram suscetíveis ao fipronil, com taxas de resistência variando de <0,5 à 2,2. Quatro populações com Rrs >1 não diferiram significativamente da cepa suscetível. Uma única população apresentou RR >2,2. No geral, o fipronil tinha sido usado principalmente para o controle de R. microplus, e em apenas três fazendas, que estavam livres de R. microplus, o fipronil era utilizado para o controle da H. irritans. Em 17 fazendas não tinha sido utilizado fipronil nos últimos três anos. Conclui-se que no Uruguai as populações de H. irritans no campo permanecem suscetíveis ao fipronil.(AU)

Acaricidal activity of (E)-cinnamaldehyde and α-bisabolol on populations of Rhipicephalus microplus (Acari: Ixodidae) with different resistance profiles.

This study aimed to investigate the acaricidal activity of (E)-cinnamaldehyde and α-bisabolol on populations of Rhipicephalus microplus with different resistance profiles. The adult immersion test (AIT) was used to characterize the susceptibility of tick populations (50 field populations) to synthetic acaricides: deltamethrin, amitraz, and chlorfenvinphos. The larval packet test (LPT) was used to determine the LC50 values for (E)-cinnamaldehyde (populations 1-25) and α-bisabolol (populations 26-50) at the concentrations of 0.31, 0.62, 1.25, 2.0, 2.5, 5.0 and 10.0 mg/mL. The susceptible strain Porto Alegre (POA) was used as a reference for calculating the resistance ratio (RR). In the AIT, deltamethrin did not show efficacy >95 % for any of the populations, whereas amitraz and chlorfenvinphos have presented efficacy >95 % for three (6 %) and 15 (30 %) populations, respectively. In the LPT, the LC50 values of (E)-cinnamaldehyde and α-bisabolol varied from 0.23 to 2.36 mg/mL and 1.57-3.01 mg/mL, respectively. The RR50 for (E)-cinnamaldehyde showed 20 (80 %) populations with values <1.0 and no population with values>1.5. As for α-bisabolol, only two (8%) populations have presented RR50 <1.0, whereas three (12 %) populations showed incipient resistance to this sesquiterpene (RR50 between 1.5 and 2.0). The results indicate that all studied tick populations showed low susceptibility to at least one of the commercial acaricides tested. In addition, comparison between the LC50 values of (E)-cinnamaldehyde and α-bisabolol for the field populations and the susceptible strain POA suggests that there is no cross-resistance of (E)-cinnamaldehyde and α-bisabolol for the tick populations evaluated, and that the differences in the LC50 values are due to population variations.