Control of Rhipicephalus microplus tick larvae in the field based on distancing from the host - Lone tick system.

The aim of this study was to evaluate a host-tick distancing control system using Rhipicephalus microplus in cattle. Its impact on the profiles of the pathogens Anaplasma marginale and Babesia bigemina was also assessed using enzyme-linked immunosorbent and real-time polymerase chain reaction. Experiments were conducted in Mato Grosso do Sul, Brazil. A total of 37 Senepol animals with natural tick infestation were used in this study. The ticks on the animals were counted, and pasture rotation was performed at 28-day intervals. This approach was repeated until the end of the experiment to maintain a low number of ticks, resulting in a final average of 6.3 ticks/host. Cattle maintained an immune response to the cattle tick fever (CTF) agents. DNA copies of A. marginale (X̄ 1.1) and B. bigemina (X̄ 0.05) did not vary significantly with the decrease in tick count throughout the study period. Based on these results, we conclude that a distancing period of 84 days between ticks and cattle in conditions of the Cerrado biome can control ticks in animals and maintain enzootic stability. Thus, it is possible to create productive breeds with lower tick control costs.

Seasonal fluctuations of Babesia bigemina and Rhipicephalus microplus in Brangus and Nellore cattle reared in the Cerrado biome, Brazil.

BACKGROUND: The tick Rhipicephalus microplus (Ixodida: Ixodidae, Canestrini, 1888) causes substantial economic and health losses in cattle production and is the main vector of Babesia bigemina (Piroplasmidae: Babesidae, Smith & Kilborne, 1893). Babesia bigemina is responsible for a tick-borne disease known as babesiosis that can cause hemolytic anemia, fever and death. In the study reported here, we investigated the relationship between the number of ticks per animal and the number of B. bigemina cytochrome b gene (cbisg) copies in the blood of Brangus and Nellore cattle reared without acaricidal treatment in the Brazilian Cerrado biome over a 1-year period. METHODS: Ticks on 19 animals (9 Brangus and 10 Nellore cattle) were counted every 18 days, and blood was collected every 36 days for 12 months. Serological samples were analyzed with an indirect enzyme-linked immunosorbent assay, and genomic DNA was analyzed by conventional PCR and quantitative PCR. The PCR products were sequenced by the Sanger method. RESULTS: The Brangus and Nellore breeds showed similar weight development and no clinical signs of babesiosis. Statistically significant differences (P < 0.05) between the breeds were observed for the number of ticks and the number of B. bigemina cbisg gene copies. CONCLUSIONS: No correlation between the number of ticks and the number of circulating copies of cbisg was observed, although Nellore cattle presented with fewer ticks than Brangus cattle and the number of cbisg copies was higher for Nellore cattle than for Brangus cattle.

Tick fauna in non-anthropogenic areas in Mato Grosso do Sul, Brazil, with the presence of the Rickettsia parkeri strain Atlantic rainforest in Amblyomma ovale.

The aim of this study was to evaluate tick fauna and identify the possible presence of Rickettsia spp. in ticks of the genus Amblyomma from two environmental preservation areas in different regions of Mato Grosso do Sul state, Brazil. CO2 traps, visual observation and cloth dragging were used to capture ticks. Three hundred ticks were submitted to the hemolymph test, and samples that showed organisms morphologically compatible with Rickettsia were used for rickettsial DNA detection by PCR. DNA was extracted using guanidine-phenol isothiocyanate, and the primers CS78 and CS323 were used for PCR, which amplified a 401-base pair fragment of the citrate synthase (gltA) gene. If positive, the DNA sample was tested by primers Rr190.70p and Rr190.602n that produce a 530 bp amplicon of the ompA gene that is present only in rickettsiae of the spotted fever group. A total of 1,745 adult ticks were collected, including 1,673 specimens of Amblyomma sculptum, 63 of Amblyomma coelebs, five of Amblyomma naponense and four of Amblyomma ovale. Thirteen ticks of the species A. ovale, A. coelebs and A. sculptum showed structures compatible with Rickettsia inside the hemocytes; after DNA extraction, the presence of Rickettsia spp. in a sample of A. ovale was confirmed by PCR in both analyzed fragments. In the sequencing analysis, 100% identity for the Rickettsia parkeri strain Atlantic rainforest was obtained according to GenBank. The two environmental preservation areas showed A. sculptum as the predominant species, as well as the presence of marked seasonality for this species. This paper is the first report of the R. parkeri strain Atlantic rainforest in A. ovale ticks in the state of Mato Grosso do Sul, Brazil.

Serological evaluation of Neospora caninum in pregnant women treated at referral center for prenatal screening in Mato Grosso do Sul, Brazil.

Neospora caninum is an obligate intracellular protozoan with canids (Canis domesticus, Canis lupus dingo, Canis latrans, Canis lupus) as its definitive hosts. The objective of this study was to detect anti-N. caninum antibodies in pregnant women seen at referral center for prenatal screening in the state of state Mato Grosso do Sul, Brazil. A total of 188 serum samples from pregnant women provided by the Instituto de Pesquisa, Ensino e Diagnósticos da APAE de Campo Grande (IPED/ APAE) were subjected to IFA test and western blot analysis. The samples were divided into three groups: 23/99 samples from the seropositive group for toxoplasmosis were positive for anti-N. caninum IgG antibodies, and 9/99 positive for IgM; in the HIV group, 7/33 were positive for IgG; and in the HIV+toxoplasmosis group, 13/56 were positive for IgG and two positive for IgM. The seropositivity for IgG was assessed by western blot by testing 43 IFA test positive samples using rNcSRS2 (Nc-p43) as antigen. The serological results of the present study suggest that exposure of these pregnant women to the parasite N. caninum and presence of IgM antibodies are indicative of recent infection. Further studies are needed to establish the possibility of active infection.

Spotted fever group Rickettsia and Borrelia sp. cooccurrence in Amblyomma sculptum in the Midwest region of Brazil.

More than 70 tick species are found in Brazil, distributed over five genera and including main vectors of infectious disease agents affecting both animals and humans. The genus Amblyomma is the most relevant for public health in Brazil, wherein Amblyomma aureolatum, Amblyomma ovale and Amblyomma sculptum have been incriminated as vectors of Rickettsia and Borrelia pathogens. The objective of this study was to investigate the presence of Rickettsia spp. and Borrelia spp. in ticks in the Brazilian mid-western savannah. DNA extraction, PCR for Borrelia spp. (flgE gene) and Rickettsia spp. (ompA and gltA genes) and subsequent sequencing were performed. A total of 1875 ticks were collected and identified as A. sculptum except for two Amblyomma coelebs ticks. Molecular evidence for Borrelia spp. and Rickettsia parkeri was found in A. sculptum. This is the first molecular evidence for R. parkeri in A. sculptum ticks in the Midwest region and Borrelia spp. circulating in a tick of the Amblyomma genus in Brazil.

Evidence of acaricide resistance in different life stages of Amblyomma mixtum and Rhipicephalus microplus (Acari: Ixodidae) collected from the same farm in the state of Veracruz, Mexico.

The objectives of this study were to evaluate the resistance of Amblyomma mixtum and Rhipicephalus microplus ticks from co-infested bovines from the Veracruz region in Mexico to different acaricide families and to demonstrate the viability of the packet test on different A. mixtum instars. The following acaricide families were used: a combination (cypermethrin 15 g + chlorpyrifos 25 g + citronella 1 g + butoxide piperonyl 15 g), amidine (formamidine 12.5 g), pyrethroid (cypermethrin 15 g), and organophosphate (dichlorvos 60 g + chlorpyrifos 20 g). Regarding the packet test in both species, resistance was found for the pyrethroid and amidine families in A. mixtum and R. microplus, as efficacy did not surpass 40 %, including in immature instars; regarding the adult immersion test in R. microplus, the efficacy was 93.3 % for the amidine family and 26.2 % for the pyrethroid family. The proposed methodology is an alternative technique to optimize resistance detection in immature ticks with a heteroxenous life cycle.

Serological evaluation of Neospora caninum in pregnant women treated at referral center for prenatal screening in Mato Grosso do Sul, Brazil / Avaliação sorológica de Neospora caninum em gestantes atendidas em centro de referência para triagem pré-natal em Mato Grosso do Sul, Brasil

Abstract Neospora caninum is an obligate intracellular protozoan with canids (Canis domesticus, Canis lupus dingo, Canis latrans, Canis lupus) as its definitive hosts. The objective of this study was to detect anti-N. caninum antibodies in pregnant women seen at referral center for prenatal screening in the state of state Mato Grosso do Sul, Brazil. A total of 188 serum samples from pregnant women provided by the Instituto de Pesquisa, Ensino e Diagnósticos da APAE de Campo Grande (IPED/ APAE) were subjected to IFA test and western blot analysis. The samples were divided into three groups: 23/99 samples from the seropositive group for toxoplasmosis were positive for anti-N. caninum IgG antibodies, and 9/99 positive for IgM; in the HIV group, 7/33 were positive for IgG; and in the HIV+toxoplasmosis group, 13/56 were positive for IgG and two positive for IgM. The seropositivity for IgG was assessed by western blot by testing 43 IFA test positive samples using rNcSRS2 (Nc-p43) as antigen. The serological results of the present study suggest that exposure of these pregnant women to the parasite N. caninum and presence of IgM antibodies are indicative of recent infection. Further studies are needed to establish the possibility of active infection.

Resumo Neospora caninum é um protozoário intracelular obrigatório que possui os canídeos (Canis domesticus, Canis lupus dingo, Canis latrans, Canis lupus) como seus hospedeiros definitivos. O objetivo deste trabalho foi detectar anticorpos anti-N. caninum em gestantes, atendidas em centro de referência para triagem pré-natal, em Mato Grosso do Sul, Brasil. Um total de 188 amostras de soro de gestantes, cedidas pelo Instituto de Pesquisa, Ensino e Diagnósticos da APAE de Campo Grande (IPED/ APAE), foram submetidas ao teste de RIFI e western blot para a detecção de anticorpos anti-N. caninum. As amostras foram divididas em três grupos: 23/99 amostras do grupo soropositivo para toxoplasmose demonstraram positividade para anticorpos IgG anti-N. caninum e 9/99 positivos para IgM. No grupo HIV 7/33 apresentaram positividade para IgG. No grupo HIV+Toxoplasmose 13/56 apresentaram positividade para IgG e duas para IgM. A soropositividade para IgG foi avaliada por western blot, testando 43 amostras positivas para RIFI, usando-se rNcSRS2 (Nc-p43) como antígeno. Os resultados sorológicos do presente estudo sugerem exposição dessas gestantes ao parasita N. caninum e a positividade para anticorpos IgM são indicativos de infecção recente. Mais estudos na área são necessários para estabelecer a possibilidade de infecção ativa.

Economic performance evaluation of Brangus and Nellore cattle breed naturally infested with Rhipicephalus microplus in an extensive production system in Central-West Brazil.

The objective of the present study was to evaluate the performance of Brangus and Nellore cattle in an extensive production system. Sixty growing bulls, 30 of the Brangus and 30 of the Nellore breeds that were naturally infested with Rhipicephalus microplus were subdivided into four groups: control Nellore (15 animals), treated Nellore (15), control Brangus (15), and treated Brangus (15). The animals in the treated groups underwent acaricidal treatments for 1 year. Tick counts, acaricidal treatments and animal weighing were performed every 18 days, and the costs of acaricidal, anti-myiasis, and preventive treatments for tick fever were recorded for cost evaluation. The treated Brangus and Nellore groups did not show a significant difference in weight gain, whereas Nellore weight gain was superior in the control groups. The cost of acaricidal treatment throughout the experimental period was $494 US. The costs of the preventive treatment for tick fever and myiasis were $98 US and $15 US, respectively. The highest rates of tick infestation were found in the control group of the Brangus animals, which served as the basis for the suggested implementation of a strategic control program for animals in the growth phase. Nellore animals showed low rates of infestation. Under the conditions of this study, the Nellore animals were more efficient than the Brangus animals because they achieved satisfactory weight gain, similar to the Brangus, and a low tick count even in the control group. This finding demonstrates that expenses related to the acaricidal, anti-myiasis and tick fever treatments are unnecessary in Nellore cattle, making these animals more profitable.

Free-living ticks in a cerrado fragment, state of Mato Grosso do Sul, Brazil / Carrapatos de vida livre em fragmento de cerrado, no Estado de Mato Grosso do Sul, Brasil

The fatal cases of spotted fever reported in some Brazilian municipalities are generally associated with the presence of ticks and capybaras (Hydrochoerus hydrochaeris). The urban forest fragments where this rodent is present are of great concern. We investigated the diversity of free ticks as well as the occurrence of ticks infected by Rickettsia in one of these fragments, in Campo Grande city, Mato Grosso do Sul state, Brazil. From winter 2012 until the autumn of 2015, using CO2 traps, we collect 1219 ticks, of which 1045 belonged to the species Amblyomma sculptum and 174 belonged to A. dubitatum. Both species were more abundant in spring (September to December) and summer (December to March). Rickettsial DNA was not detected in any of the specimens collected; however, one of the species found, A. sculptum is considered the main vector of Brazilian Spotted Fever, thus deserving constant vigilance by the public health organs.

Casos fatais de febre maculosa que foram relatados em alguns municípios brasileiros geralmente estão associados à presença de carrapatos e capivaras (Hydrochoerus hydrochaeris). Os fragmentos florestais urbanos onde este roedor está presente são de grande preocupação. Investigou-se aqui a diversidade de carrapatos livres e a ocorrência de carrapatos infectados por riquétsias em um desses fragmentos em Campo Grande, Mato Grosso do Sul. Desde o inverno de 2012 até o outono de 2015, usando armadilhas de CO2, foram coletados 1219 carrapatos, dos quais 1045 pertenciam à espécie Amblyomma sculptum e 174 pertenciam a A. dubitatum. Ambas as espécies foram mais abundantes na primavera (Setembro a Dezembro) e no verão (Dezembro a Março) . O DNA riquetsial não foi detectado em nenhum dos espécimes coletados; entretanto, uma das espécies encontradas, o A. sculptum é considerado o principal vetor da Febre Maculosa Brasileira, merecendo assim vigilância constante pelos Orgãos de Saúde Pública.

Biological parameters of Amblyomma coelebs Neumann, 1906 (Acari: Ixodidae) under experimental conditions.

One generation of Amblyomma coelebs life cycle under experimental conditions was evaluated. Ten tick pairs were allowed to feed on rabbits under laboratory conditions (LC), resulting six engorged females with a mean weight of 1,403.9 mg. Two females were maintained in a forest reserve under natural conditions (NC), and four were maintained in incubators (LC). The engorgement period lasted 10.33 days. Pre-oviposition periods were 10.75 (NC) and 22 days (LC). The mean egg-mass weight was 514.76 mg, and the blood meal conversion index was 36.67% (LC). Incubation period under NC and LC were 91 and 56.33 days and hatching rates were 50% and 28.33%, respectively. Larval engorgement period ranged from 4 to 10 days, with average weight of 1.1 mg. Engorged larvae were incubated under NC and LC, with a premolt period of 27 to 36 days and molting rate of 7.1% and 28.7%, respectively. Nymphal engorgement period ranged from 5 to 7 days, with a mean weight of 18.8 mg and a recovery rate of 54.54%. In LC, the ecdysis mean period was 24.5 days, and molting rate was 44.44%, resulting in 24 adult A. coelebs. Our results show a life cycle of 187.45 (NC) and 149 (LC) days.

Biological parameters of Amblyomma coelebs Neumann, 1906 (Acari: Ixodidae) under experimental conditions / Parâmetros biológicos de Amblyomma coelebs Neumann, 1906 (Acari: Ixodidae) em condições experimentais

Abstract One generation of Amblyomma coelebs life cycle under experimental conditions was evaluated. Ten tick pairs were allowed to feed on rabbits under laboratory conditions (LC), resulting six engorged females with a mean weight of 1,403.9 mg. Two females were maintained in a forest reserve under natural conditions (NC), and four were maintained in incubators (LC). The engorgement period lasted 10.33 days. Pre-oviposition periods were 10.75 (NC) and 22 days (LC). The mean egg-mass weight was 514.76 mg, and the blood meal conversion index was 36.67% (LC). Incubation period under NC and LC were 91 and 56.33 days and hatching rates were 50% and 28.33%, respectively. Larval engorgement period ranged from 4 to 10 days, with average weight of 1.1 mg. Engorged larvae were incubated under NC and LC, with a premolt period of 27 to 36 days and molting rate of 7.1% and 28.7%, respectively. Nymphal engorgement period ranged from 5 to 7 days, with a mean weight of 18.8 mg and a recovery rate of 54.54%. In LC, the ecdysis mean period was 24.5 days, and molting rate was 44.44%, resulting in 24 adult A. coelebs. Our results show a life cycle of 187.45 (NC) and 149 (LC) days.

Resumo Uma geração do ciclo de vida de Amblyomma coelebs em condições laboratoriais foi avaliada. Dez casais de carrapatos foram alimentados em coelhos sob condições laboratoriais (CL), resultando em seis fêmeas ingurgitadas, com um peso médio de 1.403,9 miligramas (mg). Duas fêmeas foram mantidas em uma reserva florestal sob condições naturais (CN), e quatro foram mantidas em incubadoras (CL). O período de ingurgitamento durou 10,33 dias. Períodos de pré-postura foram de 10,75 (CN) e 22 dias (CL). O peso médio das massas de ovos foi de 514,76 mg e o índice de conversão alimentar foi de 36,67% (CL). O período de incubação em CN e CL foi de 91 e 56,33 dias e os percentuais de eclosão foram de 50% e 28,33%, respectivamente. O período de ingurgitamento larval variou de quatro a 10 dias, com peso médio de 1,1 mg. Larvas ingurgitadas foram incubadas em CN e CL, com período de pré-muda de 27 a 36 dias e percentual de ecdise de 7,1% e 28,7%, respectivamente. O período de ingurgitamento das ninfas oscilou de cinco a sete dias, com peso médio de 18,8 mg e uma taxa de recuperação de 54,54%. Em CL, o período médio de ecdise foi de 24,5 dias, e o percentual de muda foi 44,44%, resultando em 24 adultos de A. coelebs. Estes resultados demonstram um ciclo de vida de 187,45 (CN) e 149 (CL) dias.

Efficacy profile of Cypermethrin and Chlorpyrifos based acaricides on Rhipicephalus microplus control on cattle in the rearing phase, naturally infested and exposed to tick fever agents in central Brazil.

The objective of this work was to evaluate the efficacy of two cypermethrin- and chlorpyrifos-based acaricides in controlling Rhipicephalus microplus in a naturally infested bovine herd and in in vitro tests, as well as to monitor the animals for tick fever. Male bovines in the rearing phase were used, with 30 Brangus and 30 Nellore animals naturally infested. The groups were composed as follows: 15 Nellore treated, 15 Nellore control, 15 Brangus treated and 15 Brangus control. Every 18 days, the animals were monitored for tick count, acaricide treatment, weight, blood pack cell volume, and clinical signs. For in vitro tests, the larval packet test, adult immersion test and DNA amplification for tick fever diagnosis were performed. In the first animal treatment period, product 1 (cypermethrin, 15 g + chlorpyrifos, 25 g + citronellal, 1 g) was used; in the second period, product 2 (cypermethrin, 15 g + chlorpyrifos, 30 g + fenthion, 15 g) was used. In Brangus animals, the mean efficacy was 35.1% and 95.8% in the first and second periods, respectively, for the same animals. For Nellore animals, the efficacy in periods one and two was 51% and 97.1%, respectively. The in vitro results showed efficacy above 95% for the two challenged acaricides. The Brangus animals showed a high production of ticks associated with the presence of tick fever agents, which could generate risks for the disease's enzootic stability.

Evaluation of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) resistance to different acaricide formulations using samples from Brazilian properties.

The Rhipicephalus (Boophilus) microplus tick is responsible for considerable economic losses in Brazil, causing leather damage, weight loss and reduced milk production in cattle and results in the transmission of pathogens. Currently, the main method for controlling this tick is using acaricides, but their indiscriminate use is one of the major causes of resistance dissemination. In this study, the adult immersion test (AIT) was used to evaluate resistance in ticks from 28 properties located in five different states (Mato Grosso do Sul, Mato Grosso, Ceará, São Paulo, e Minas Gerais) and the Distrito Federal (DF) of Brazil. The resistance was found in 47.64% of the repetitions demonstrating an efficacy of less than 90% in various locations throughout the country. The larvae packet test was used to evaluate samples from ten properties in four states (Mato Grosso do Sul, Mato Grosso, Minas Gerais and Espírito Santo). Spray products belonging to the main classes of acaricides, including combination formulations, were used in both types of test. New cases of resistance were found on properties within the states of Ceará, Espírito Santo and Mato Grosso, where such resistance was not previously reported.

Evaluation of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) resistance to different acaricide formulations using samples from Brazilian properties / Avaliação da resistência de Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) a diferentes formulações acaricidas utilizando amostras de propriedades brasileiras

Abstract The Rhipicephalus (Boophilus) microplus tick is responsible for considerable economic losses in Brazil, causing leather damage, weight loss and reduced milk production in cattle and results in the transmission of pathogens. Currently, the main method for controlling this tick is using acaricides, but their indiscriminate use is one of the major causes of resistance dissemination. In this study, the adult immersion test (AIT) was used to evaluate resistance in ticks from 28 properties located in five different states (Mato Grosso do Sul, Mato Grosso, Ceará, São Paulo, e Minas Gerais) and the Distrito Federal (DF) of Brazil. The resistance was found in 47.64% of the repetitions demonstrating an efficacy of less than 90% in various locations throughout the country. The larvae packet test was used to evaluate samples from ten properties in four states (Mato Grosso do Sul, Mato Grosso, Minas Gerais and Espírito Santo). Spray products belonging to the main classes of acaricides, including combination formulations, were used in both types of test. New cases of resistance were found on properties within the states of Ceará, Espírito Santo and Mato Grosso, where such resistance was not previously reported.

Resumo O carrapato Rhipicephalus (Boophilus) microplus é responsável por consideráveis perdas econômicas no Brasil, causando, nos bovinos, danos ao couro, perda de peso, redução na produção de leite, o que resulta na transmissão também de patógenos. Atualmente, o principal método de controle para esse carrapato é o uso de acaricidas, porém seu uso indiscriminado é uma das principais causas da disseminação da resistência. Neste estudo, o teste de imersão de adultos (TIA) foi utilizado para avaliar a resistência em carrapatos provenientes de 28 propriedades, localizadas em cinco diferentes estados brasileiros (Mato Grosso do Sul, Mato Grosso, Ceará, São Paulo, e Minas Gerais) e no Distrito Federal (DF). A resistência foi encontrada em 47,64% das repetições, demonstrando uma eficácia de menos de 90% em vários lugares do país. O teste de pacote de larva foi utilizado para avaliar amostras de 10 propriedades em quatro estados (Mato Grosso do Sul, Mato Grosso, Minas Gerais e Espírito Santo). Produtos de pulverização, pertencentes às principais classes de acaricidas, incluindo-se associações, foram utilizados em ambos os testes. Novos casos de resistência foram encontrados em propriedades nos estados do Ceará, Espírito Santo e Mato Grosso, nas quais a resistência ainda não havia sido previamente reportada.