What is the optimal timing to initiate strategic control of Rhipicephalus microplus in taurine cattle in a tropical region?

Cattle tick control poses a significant challenge for livestock in tropical and subtropical regions. The objective of this study was to determine the most suitable timing to initiate a strategic tick control program and to identify the ideal number of acaricide treatments for adult taurine cattle (Bos taurus taurus) in a tropical region throughout the year. Three groups with 10 bovines each were performed: T01 (strategic treatment in late autumn/winter/late spring, every 28 days), T02 (strategic treatment to act in the "first tick generation" - early spring/summer/early autumn, every 28 days) and T03 (control). Tick counts (females 4-8 mm) were conducted every 14 days. If the tick burden in any group reached 30 or more during these counts, we applied an additional treatment. Over the course of a year, T02 required significantly fewer (p < 0.05) acaricide treatments than T01, with nine treatments for T02 and eleven for T01. Furthermore, during the tick counts, animals in T02 showed a lower tick burden compared to those in T01. Initiating the strategic tick control program in early spring, corresponding to the first tick generation, proved more effective than starting in autumn. This approach not only required fewer acaricide treatments but also resulted in a reduced tick burden. These benefits are particularly valuable in terms of animal welfare and managing acaricide resistance issues.

Susceptibility of Amblyomma sculptum, Vector of Rickettsia rickettsii, Ticks from a National Park and an Experimental Farm to Different Synthetic Acaricides.

Amblyomma sculptum is a relevant tick species from a One Health perspective, playing an important role as a vector of Rickettsia rickettsii, the main agent of spotted fever rickettsiosis in Brazil. In this study, we evaluated the susceptibility of two A. sculptum populations from Goiás state (midwestern Brazil) to different acaricides. The first tick population (GYN strain) originated from an experimental farm, where the ticks are annually exposed to acaricides. The second (PNE strain) was collected in a national park (Emas National Park), where the ticks had not been exposed to acaricides. Immersion tests were conducted with 21-day-old laboratory-reared larvae and nymphs originating from adult ticks collected in the areas mentioned above. The chosen acaricides were two synthetic pyrethroids (cypermethrin and deltamethrin), one organophosphate (chlorfenvinphos), one formamidine (amitraz), and two combinations of pyrethroids and organophosphates (cypermethrin, chlorpyrifos and citronellal; cypermethrin, fenthion and chlorpyrifos). Mortality data were used to determine the lethal concentration (LC) values at which 50%, 90%, and 99% of the ticks died (LC50, LC90, and LC99, respectively), and resistance ratios (RR) were calculated based on the LC values. The RR revealed differences between the acaricide-exposed (GYN) and unexposed (PNE) tick strains. The PNE strain larvae and nymphs were susceptible to all the tested acaricides. The GYN strain larvae were tolerant to cypermethrin, whereas the nymphs were tolerant to deltamethrin, chlorfenvinphos, and the combination of cypermethrin, chlorpyrifos, and citronellal (2 < RR ≤ 10). The GYN strain nymphs were resistant to amitraz (RR > 10). This is the first report of A. sculptum nymphs with resistance to amitraz and tolerance to deltamethrin, chlorfenvinphos, and the combination of cypermethrin, chlorpyrifos, and citronellal.

Effect of a preventive strategic control program, with imidocarb dipropionate, against tick fever agents in dairy calves.

Chemoprophylaxis with dipropionate imidocarb (IMD) is a method adopted to prevent cattle tick fever (TF). Sixty weaned dairy heifers (±60 days old), without previous exposure to Rhipicephalus microplus ticks, were housed in Tifton paddocks and were subsequently exposed to R. microplus ticks and monitored up to 315 days old. Thirty animals were kept as controls (T01) and 30 received five preventive strategic treatments with IMD at 21-day intervals (T02). The heifers were monitored weekly by means of packed cell volume (PCV) and blood smears to evaluate the presence of TF agents. Salvage treatments (ST) with diminazene and enrofloxacin were administered when animals showed PCV ≤ 24%. The A. marginale prevalence was 39.3% and 37.7%, B. bovis 6.0%, and 7.3%, and B. bigemina 16.3% and 13.7% for T01 and T02, respectively. Regarding PCV values, group T01 showed lower PCV than group T02, between 119 and 161 days of life, but when animals were 196, 210, 217, and between 252 to 301 days old, an inversion occurred. The IMD treatment protocol was effective in group T02 from day 91 to 175 while treatment was being administered, but from day 182 to 315 after the IMD treatment protocol was completed, the number of salvage treatments against TF agents performed in T02 group increased significantly. The sequential application of IMD treatments with intervals less than 21 days is not recommended.

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.

Population dynamics of Rhipicephalus microplus in dairy cattle: influence of the animal categories and correlation with milk production.

Is well known the taurine and zebuine susceptibility to Rhipicephalus microplus. Few are the reports regarding tick population dynamics between the same herd/breed, and because of this, two experiments were performed. In the 1st, the cattle tick population dynamics in dairy nursing calves (reared collective and individually), weaning calves (4-16 months), heifers (17-29 months), cows in lactation and dry cows (≥ 30 months) from the same herd, tick burden and milk production correlation were performed, for two years. R. microplus females (4.5-8.0 mm) counts and the milk production were performed every 28 and 14 days, respectively. In the 2nd experiment, bovines belonging to different categories/age (newborn without previous contact with tick; 12-13 months with tick contact since birth; and 23-24 months with tick contact since birth) were experimentally infested with 30,000 R. microplus larvae, to quantify the number of fully engorged females detached from these animals. In the 1st experiment, when the mean counts of tick were ≥ 30 all animals of the group were treated. Nursing calves showed 3-4 peaks of ticks, animals reared individually showed smaller (p ≤ 0.05) tick burden than those reared collectively. Weaning calves (4-8 months) showed 5 tick peaks/year and higher mean tick burden was found than other categories. On the other hand, animals with 17-29 months of age showed smallest (p ≤ 0.05) tick burden, with 3 tick peaks/year. When the animal become lactating the tick burden increase, and 5 peaks/year occurred, and decrease again in dry cows (p ≤ 0.05) showing 4-5 tick peaks/year. Weaning calves and lactating cows received more acaricide treatments (p ≤ 0.05), 18 and 15, respectively. Nursing calves reared individually, and heifers (21-29 months) were the categories that received two acaricide treatments. The more milk the cow produce, more ticks it has (p ≤ 0.05). In the 2nd experiment, more (p ≤ 0.05) fully engorged females were recovered from younger animals than older ones. So, different tick control strategies need to be adopted in different dairy cattle categories, and the tick burden should be considered, once the effect may be more inherent to the animal rather than the strategy adopted.

Tick infestation level interferes with spray formulation (organophosphate + pyrethroid) efficacy against Rhipicephalus microplus.

Due to the scarcity of information regarding the control of Rhipicephalus microplus on bovines with different tick burdens, tick counts and the efficacy of a commercial spray formulation (aspersion bath with organophosphate + pyrethroid) were evaluated for animals with a low and high tick burdens for 35 days in a stall test. Thirty-two crossbreed Bos taurus indicus x Bos t. taurus experimentally infested with R. microplus larvae were divided into four groups: T01 high infestation control, T02 high infestation treated, T03 low infestation control, and T04 low infestation treated. The bovines were kept individually in pens and R. microplus females were collected daily. All data were statistically analyzed with a significance level of 5% (P ≤ 0.05). For linear regressions, variables with P≤0.05 and the highest coefficients of determination (R2 ≥ 0.70) were considered the best descriptors. The reliability level was 95%. Tick counts differed (P<0.0001) between T01 and T02 (high infestation control vs. treated), except at day 2 (P >0.05) post-treatment, and efficacy ranged from 85.2 to 50.6%. The number of collected engorged females was lower (P<0.0001) for T04 than for T03 (low infestation treated vs. control) animals from day 3 to 29, and efficacy ranged from 95.2 to 69.8%. In addition, tick burden and efficacy were negatively correlated for both the group of animals with a high tick burden (r = -0.5256; p = 0.0012; R² = 0.2762) and the group of animals with a low tick burden (r = -0.9817; p < 0.0001; R² = 0.9638). In conclusion, a high tick burden on bovines decreases the efficacy of the tested spray acaricide.

Strategic control of cattle co-parasitized by tick, fly and gastrointestinal nematodes: Is it better to use ecto + endoparasiticide or just endectocide formulations?

Ticks, flies, and gastrointestinal helminths (GINs) significantly affect cattle productivity; thus, ectoparasiticide, endoparasiticide, and endectocide drugs have commonly been used for their control. The study aimed to compare the technical (parasites counts), productive, and financial effects of a treatment protocol comprising ecto- + endoparasiticides formulations (T01: fluazuron 2.5 mg/kg + fipronil 1.25 mg/kg and fenbendazole 5 mg/kg; n = 15) to a treatment with one formulation of endectocide (T02: ivermectin 450 µg/kg + abamectin 250 µg/kg; n = 15) over 308 days under field conditions in crossbred cattle co-parasitized by Rhipicephalus microplus, Haematobia irritans, and GINs. Bovine weight gain and return on investment (ROI) were also evaluated. Bovines from T01 received four treatments against the cattle tick and two against two GINs. For T02, four treatments were performed. Animals from T01 gained 15.4 kg more than T02 and provided a comparative ROI of 15.8. In cattle co-parasitized with R. microplus, H. irritans, and GINs, the treatment protocol used in this study with ecto- + endoparasiticidal action formulations showed better technical results regarding parasite counts and productive and financial data than the strategic treatment protocol using only an endectocide formulation.

Rhipicephalus microplus seasonal dynamic in a Cerrado biome, Brazil: An update data considering the global warming.

The present study evaluated over two years the seasonal dynamics of Rhipicephalus microplus in a Cerrado biome of midwestern Brazil (Goiânia, Goiás) and correlated the current climatic conditions (environmental temperature, insolation, rainfall, relative humitidy and saturation deficit of the atmosphere) of this site with 30 years ago. In addition, the data collected in the present research were compared with climatic conditions data and R. microplus population dynamics conducted by our group in other regions (Formiga, Minas Gerais and Jaboticabal, São Paulo) within the same biome. For the parasitic phase dynamics, R. microplus females (4.5-8.0 mm) counts were performed on tick natural infested cows kept in Panicum maximum paddocks. To verify the larvae dynamics on pasture the flannel dragg technique on natural infested pasture was performed, and the height of the grass was measured. Five peaks of engorged R. microplus females on animals and larvae infestations on pastures were observed. Being three and two peaks during the rainy and dry season of the year, respectively. Insolation (r = 0.8758; P = 0.00009; R²â€¯= 0.7670), rainfall (r = -0.8572; P = 0.0002; R²â€¯= 0.7348), maximum environmental temperature in summer (r = 0.9999; P < 0.0087; R²â€¯= 0.9988) and the saturation deficit of the atmosphere in autumn (r = -0.9789; P = 0.0211; R²â€¯= 0.9582) influenced the seasonal dynamics of R. microplus larvae on pastures. While the forage height did not influence the number of larvae found on pasture (r = 0.1545; P = 0.7682; R²â€¯= 0.0239). The comparison between the climatic data of the last 30 years with the current climatological data (2012 to 2019) in the state of Goiás, and the comparison with R. microplus population dynamics from other two locations in the same biome indicated that the increase of the environmental temperature due to the global warming possibly is a determining factor which increase the number of one or up two tick annual peaks. In addition, this climatic variable was responsible for increase the population density of ticks on pastures and animals observed in July/winter of the present study, in comparison to other regions with the same biome, as discussed in this work.

Evaluation of different strategic control protocols for Rhipicephalus microplus on cattle according to tick burden.

The development of a strategic protocol is fundamental to the control of Rhipicephalus microplus. The efficacy of different strategic tick control protocols (Day 1 to Day 119) using commercial acaricides according to R. microplus burden on bovines was evaluated. Female ticks (4.5-8 mm in length) were initially counted on cattle to form four groups of bovines with low and high tick burdens (treated and control), with 10 animals each. The animals were then (Day 0) kept in different paddocks, and the bovines from the two treated groups received their first treatment. The main difference in the protocols was the use of a spray formulation on animals with high tick burden. Additionally, the criterion for animal re-treatment (treatment interval) during the proposed strategic protocol was evaluated according to tick length by visual inspection - if 30% of the bovines of a group presented ticks < 4 mm between their legs and dewlap, the treatment was applied, which could change the originally proposed intervals. The number of days for which animals from the treated groups remained without acaricide administration (palliative control, ≥30 tick females < 4 mm) after the end of the strategic tick control protocol was recorded. Tick counts and visual inspection were done weekly until days 203 and 119, respectively. The results showed that the adopted protocol was successful based on calculated efficacy. Mean efficacies from Day 7 to Day 119 were 86.1% and 96.1% for treated groups with low and high tick burdens, respectively, and mean tick count did not differ between them (P > 0.05) but did differ from their respectively controls (P = <0.0001). The use of spray formulation at the beginning of the protocol for animals with a high tick burden helped to avoid harmful tick infestations. Visual analysis of tick length anticipated the proposed treatment intervals. Treated groups remained without palliative treatment for 70 days, while control groups needed treatment every 28 days. It was concluded that strategic control protocols must be developed according to tick burden, and tick length can be used to indicate when treatment needs to be applied.

An update on amitraz efficacy against Rhipicephalus microplus after 15 years of disuse.

Amitraz is an acaricide that is widely used in veterinary medicine to control the cattle tick Rhipicephalus microplus. However, controversy exists in the literature regarding the resistance of R. microplus to this product. The present work provides an update on the acaricidal efficacy of amitraz (Triatox®, 12.5 % amitraz) after 15 years without its use on a property. Two in vivo (bovines treated with amitraz and submitted to tick counts, n = 20 animals) and one in vitro (adult immersion test, n = 40 ticks) assays were performed to determine product efficacy. The efficacy of the commercial formulation tested in the first in vivo trial ranged from 14.1 to 47.0%, and in the second from 3.6 to 35.1%, for the 28 days of the experiments. Efficacy for the in vitro trial was 47.38%. The dose recommended by the manufacturer of the product did not cause mortality to most of the ticks of this strain, and efficacy/resistance was not reverted or modified after 15 years (estimated 60 tick generations).

Does bathing affect tick and flea burdens and ectoparasiticide effectiveness of a spot-on formulation (fipronil + (S)-methoprene) for dogs?

This work evaluated tick and flea burdens and the efficacy of a single topical application of 10% fipronil + 9% (S)-methoprene spot-on against experimental infestations of Rhipicephalus sanguineus sensu lato and Ctenocephalides felis felis on dogs that were submitted to regular baths for 56-days post-treatment. Four treatments (n = 24) were evaluated: no chemical treatment and no bathing (T01); no chemical treatment + bathing (T02); chemical treatment and no bathing (T03) and chemical treatment + bathing (T04). Dogs were infested with adult ticks and fleas seven days pre-treatment (-7). The ectoparasites were then counted and removed on day -5, followed by a new infestation on day -2 and treatment on day 0. Ticks and fleas were then counted without removal on day 1 and counted with removal on day 2. The dogs were then repeatedly infested with both ectoparasites on days 5, 12, 19, 26, 33, 40, 47 and 54 post-treatment. Baths and ectoparasite counts with removal were performed every 48 h before and after experimental infestation, respectively. Animals of T04 had lower (P ≤ 0.05) tick burden than animals from T01 and T02 from day 2 to day 28. For fleas, the counts varied among days of the study. Dogs from T03 presented lower (P ≤ 0.05) flea burden than dogs from T01 from day 1 until day 56. The same was true for T04 when compared to T01 from day 1 to day 42. Animals of T02 greater (P ≤ 0.05) flea burden than animals of T04 in all dates post treatment. In addition, dogs from T02 presented higher (P ≤ 0.05) flea burden than T01 from day 7 to day 28 and from day 49 to day 56. In sum, the efficacy of the spot-on formulation used declined over the time against both ectoparasites. The efficacy for ticks ranged from 0.0 to 96.5% and 0.0 to 98.52% for T03 and T04, respectively. While for fleas, the efficacy of the formulation ranged from 67.41 to 100% for T03 and 40.18 to 100% for T04. So, bathing dogs that were not treated increased C. f. felis burden but not R. sanguineus s. l. burden. The residual efficacy of the product had a shorter duration against these ectoparasites for dogs that received subsequent bathing.

Potential of entomopathogenic nematodes of the genus Heterorhabditis for the control of Stomoxys calcitrans (Diptera: Muscidae) / Potencial de nematoides entomopatogênicos do gênero Heterorhabditis para o controle de Stomoxys calcitrans (Diptera: Muscidae)

Abstract The present study verified the pathogenic potential of entomopathogenic nematodes (EPNs) of the genus Heterorhabditis (Heterorhabditis bacteriophora, isolate HP88 and Heterorhabditis baujardi isolate LPP7) to immature stages of Stomoxys calcitrans in the laboratory. All EPN concentrations of the H. bacteriophora HP88 strain caused mean larval mortality greater than 90% after four days. Higher concentrations of the H. baujardi LPP7 isolate (≥50 EPNs/larva) eliminated more than 70% of larvae after six days with the concentration 200 EPNs/larva reaching mortality levels of 93.3%. The larval mortality at all concentrations of EPNs (25, 50, 100, 150, and 200 EPNs/larva) for both strains was significant (p<0.05) when compared to the respective control groups. Concentrations of H. bacteriophora HP88 yielded an LC50 of 0.36 EPN/larva and LC90 of 29.1; while H. baujardi LPP7 yielded an LC50 of 39.85 and LC90 of 239.18. H. bacteriophora HP88 provided greater inhibition of the emergence of adults when compared to the response obtained with H. baujardi LPP7. EPNs did not cause considerable mortality when applied directly to pupae. The set of observed results suggests that the EPNs of the genus Heterorhabditis, isolates HP88 and LPP7, are a promising alternative in the control of the stable fly.

Resumo O presente estudo verificou o potencial patogênico de nematoides (NEP) do gênero Heterorhabditis (H. bacteriophora - isolado HP88 e H. baujardi - isolado LPP7) para estágios imaturos de Stomoxys calcitrans em laboratório. Todas as concentrações de NEPs da cepa HP88 causaram mortalidade larval média maior que 90% após quatro dias. Concentrações mais elevadas da cepa LPP7 (≥50 NEPs/larva) eliminaram mais de 70% após seis dias, com a concentração 200 NEPs/larva atingindo níveis de 93,3%. A mortalidade larval em todas as concentrações de NEPs (25, 50, 100, 150, and 200 NEPs/larva) para ambas as cepas foi significativa (p<0,05), quando comparadas aos respectivos grupos controle. Concentrações de H. bacteriophora HP88 obtiveram LC50 de 0,36 NEP/larva e LC90 de 29,1; enquanto H. baujardi LPP7 obteve LC50 de 39,85 e LC90 de 239,18. O isolado H. bacteriophora HP88 propiciou maior inibição da emergência de adultos, quando comparado à resposta obtida com H. baujardi LPP7. NEPs não provocaram mortalidade considerável quando aplicados diretamente sobre pupas. O conjunto de resultados observados indica os nematoides entomopatogênicos do gênero Heterorhabditis (estirpes HP88 e LPP7) como uma alternativa promissora no controle da mosca dos estábulos.

Potential of entomopathogenic nematodes of the genus Heterorhabditis for the control of Stomoxys calcitrans (Diptera: Muscidae).

The present study verified the pathogenic potential of entomopathogenic nematodes (EPNs) of the genus Heterorhabditis (Heterorhabditis bacteriophora, isolate HP88 and Heterorhabditis baujardi isolate LPP7) to immature stages of Stomoxys calcitrans in the laboratory. All EPN concentrations of the H. bacteriophora HP88 strain caused mean larval mortality greater than 90% after four days. Higher concentrations of the H. baujardi LPP7 isolate (≥50 EPNs/larva) eliminated more than 70% of larvae after six days with the concentration 200 EPNs/larva reaching mortality levels of 93.3%. The larval mortality at all concentrations of EPNs (25, 50, 100, 150, and 200 EPNs/larva) for both strains was significant (p<0.05) when compared to the respective control groups. Concentrations of H. bacteriophora HP88 yielded an LC50 of 0.36 EPN/larva and LC90 of 29.1; while H. baujardi LPP7 yielded an LC50 of 39.85 and LC90 of 239.18. H. bacteriophora HP88 provided greater inhibition of the emergence of adults when compared to the response obtained with H. baujardi LPP7. EPNs did not cause considerable mortality when applied directly to pupae. The set of observed results suggests that the EPNs of the genus Heterorhabditis, isolates HP88 and LPP7, are a promising alternative in the control of the stable fly.

Virulence of entomopathogenic nematodes (Rhabditida: Steinernematidae, Heterorhabditidae) to spittlebug Mahanarva spectabilis (Hemiptera: Cercopidae) / Virulência de nematoides entomopatogênicos (Rhabditida: Steinernematidae, Heterorhabditidae) para cigarrinha-das-pastagens Mahanarva spectabilis (Hemiptera: Cercopidae)

The spittlebug can seriously limit the production of forage, and the use of chemical products to control insect pests is costly and can harm the environment. Thus, there is a need to reduce the reliance on chemical agents by developing new strategies. The virulence of nine strains of entomopathogenic nematodes (EPNs) to spittlebugs was investigated under laboratory and greenhouse conditions. Fourth/fifth-instar nymphs of Mahanarva spectabilis were exposed to EPNs in the laboratory and the most virulent strains were applied on the nymphs in the greenhouse at concentrations of 2,000 and 4,000 EPNs/mL. The efficacy of the pathogenic agent was confirmed by the dissection of dead hosts. All the tested strains were pathogenic to the M. spectabilis nymphs in laboratory, particularly Steinernema carpocapsae, S. feltiae, S. riobrave and Heterorhabditis amazonensis RSC1, each of which caused nymph mortality higher than 80%. The concentration did not influence the efficiency of the strains, and those selected in the laboratory had similar efficiency in the greenhouse, except for S. carpocapsae, which was not as effective as the others. Entomopathogenic nematodes can be included in integrated pest management programs to M. spectabilis.(AU)

A cigarrinha-das-pastagens pode causar sérias limitações na produção de forragens, e o uso de produtos químicos para o controle é caro, além de poder prejudicar o ambiente. Então, existe a necessidade de reduzir o uso de agentes químicos através do desenvolvimento de novas estratégias de controle dessa praga. A virulência de nove isolados de nematoides entomopatogênicos (NEPs) sobre a cigarrinha-das-pastagens foi avaliada em condições de laboratório e casa-de-vegetação. Ninfas de quarto/quinto ínstar de Mahanarva spectabilis foram expostas aos isolados de NEPs em laboratório, e os mais virulentos foram aplicados sobre as ninfas em casa-de-vegetação sob as concentrações de 2000 e 4000 JIs/mL. A eficácia do agente patogênico foi confirmada pela dissecação dos hospedeiros mortos. Todos os isolados testados foram patogênicos às ninfas da cigarrinha-das-pastagens em laboratório, particularmente Steinernema carpocapsae, S. feltiae, S. riobrave e Heterorhabditis amazonensis RSC1, cada um deles causando mortalidade maior que 80%. A concentração não influenciou a eficiência, exceto para S. carpocapsae, o qual não foi tão efetivo como os demais em casa-de-vegetação. Nematoides entomopatogênicos podem ser incluídos em programas de manejo integrado de M. spectabilis.(AU)

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

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

Compatibilidade de Heterorhabditis amazonensis (Rhabditida: Heterorhabditidae) isolado RSC-5 com diferentes carrapaticidas utilizados no controle de Rhipicephalus microplus (Acari: Ixodidae) / Compatibility of Heterorhabditis amazonensis (Rhabditida: Heterorhabditidae) strain RSC-5 with different acaricides used for the control of Rhipicephalus microplus (Acari: Ixodidae)

O estudo teve como objetivo avaliar a viabilidade de Heterorhabditis amazonensis isolado RSC-5 após exposição a diferentes carrapaticidas utilizados no controle de Rhipicephalus microplus. Foram constituídos seis tratamentos, cada um composto por um produto, sendo cada grupo com 75.000 nematoides (NEPs) em suspensão de 20 mL de solução de diferentes carrapaticidas, em concentração comercial. O controle foi formado por 75.000 NEPs e 20 mL de água destilada, e todos os grupos foram mantidos em câmara climatizada a 25°C. A avaliação do percentual de sobrevivência e infectividade em lagartas Galleria mellonella foi realizada 24 e 72 horas após o início do experimento. A mortalidade de lagartas no teste de infectividade foi analisada após 72 e 120 horas. Com 24 horas de exposição, o percentual de sobrevivência de H. amazonensis RSC-5 não foi significativamente reduzido (p > 0,05) somente na exposição ao princípio ativo deltametrina. O mesmo foi observado no período de 72 horas em relação à associação clorpirifós + cipermetrina + butóxido de piperonila + citronelal. Não foi constatada sobrevivência de nenhum juvenil infectivo nos grupos expostos à associação clorfenvinfós + diclorvós. A exposição ao clorfenvinfós resultou em percentual de sobrevivência inferior a 50% após 72 horas. O potencial de infectar lagartas de G. mellonella foi reduzido apenas no grupo tratado com o princípio ativo clorfenvinfós. Dessa forma, é possível concluir que o princípio ativo clorfenvinfós e a associação clorfenvinfós + diclorvós não foram compatíveis com H. Amazonenses RSC-5, causando redução no percentual de sobrevivência e infectividade dos juvenis desse nematoide. Os outros produtos foram compatíveis, não causando redução na infectividade do isolado testado.(AU)

The aim of this study was to assess the viability of Heterorhabditis amazonensis strain RSC-5 after exposure to different acaricides used for Rhipicephalus microplus control. Six treatment groups were formed, one for each product. Each group was composed of 75,000 nematodes in a 20 mL solution of different acaricides, at commercial concentration. The control group was formed by the same number of nematodes in 20 mL of distilled water. All the groups were kept in a climate-controlled chamber at 25°C. The percentage of survival and infectivity in Galleria mellonella caterpillars were determined 24 and 72 hours after the beginning of the experiment. The mortality of the caterpillars in the infectivity test was assessed 72 and 120 hours. After 24 hours of exposure, only the active ingredient deltamethrin did not significantly reduce the survival percentage of H. amazonensis RSC-5 (p > 0.05). The same was observed after 72 hours of exposure to the combination of chlorpyriphos + cypermethrin + piperonyl butoxide + citronellal. There was no survival of infective juveniles in the groups exposed to the combination of chlorphenvinphos + dichlorvos. The exposure to chlorphenvinphos for 72 hours resulted in 50% of mortality. The potential to infect G. mellonella caterpillars was only reduced in the group treated with the active ingredient chlorphenvinphos. Chlorphenvinphos and the combination of chlorphenvinphos + dichlorvos were not compatible with H. amazonensis RSC-5, causing a reduction in the survival and infectivity of juveniles of this nematode, while the other products were compatible, causing no reduction in the infectivity of this isolate.(AU)

A new approach to characterization of the resistance of populations of Rhipicephalus microplus (Acari: Ixodidae) to organophosphate and pyrethroid in the state of Minas Gerais, Brazil.

The monitoring of resistance of cattle tick populations in Brazil to the chemical bases in use is largely limited to investigation of the phenotypic profile. There are few studies investigating the role played by the genotypic profile in acaricide resistance in the country. Therefore, the aim of the present study was to carry out molecular characterization and trace out the genetic profile of populations of Rhipicephalus microplus with respect to resistance to the organophosphate and pyrethroid chemical groups. For that purpose, larvae were genotyped belonging to 587 populations for pyrethroids and 306 for organophosphates, using the polymerase chain reaction technique. It was found that 75.49% and 97.44% of the larvae studied showed resistance to the organophosphates and pyrethroids, respectively. Among the populations resistant to pyrethroids, 91.9% were heterozygotes, showing that most of the resistant populations have only one allele responsible for resistance. Therefore, it is possible to conclude that the genotyped populations have high resistance to organophosphates, and even more so to pyrethroids. This information is fundamental for understanding the mechanisms of resistance of R. microplus to acaricides, to enable improvement of control techniques.

Chemical composition and acaricidal activity of essential oil from Lippia sidoides on larvae of Dermacentor nitens (Acari: Ixodidae) and larvae and engorged females of Rhipicephalus microplus (Acari: Ixodidae).

The aim of this work was to identify the compounds and to investigate the acaricidal activity of the essential oil from the leaves of Lippia sidoides on Rhipicephalus microplus and Dermacentor nitens. The oil was obtained by hydrodistillation and analyzed by gas chromatography (GC/FID) and gas chromatography/mass spectrometry. In total, 15 compounds comprising 99.97 % of the total peak area were identified. The main constituent of the essential oil was thymol (67.60 %). The acaricidal activity was assessed by the modified larval packet test, with oil concentrations of 2.5, 5.0, 10.0, 15.0, and 20.0 µl/ml, and by the female immersion test with concentrations of 10.0, 20.0, 40.0, 60.0, and 80.0 µl/ml. The mortality of the R. microplus and D. nitens larvae was greater than 95 % starting at concentrations of 10.0 and 20.0 µl/ml, respectively. In the test with the engorged females, the L. sidoides essential oil starting at a concentration of 40.0 µl/ml caused a significant reduction (p < 0.05) in the values of the egg mass weight and egg production index. The viability of the eggs was affected in all the treated groups, with significantly lower hatching rates (p < 0.05) in relation to the control group. The control percentages at concentrations of 10.0, 20.0, and 30.0 µl/ml were 54, 57, and 72 %, and reached 100 % at the highest two concentrations (60.0 and 80.0 µl/ml). Therefore, it can be concluded that the essential oil from the leaves of L. sidoides has acaricidal activity on R. microplus and D. nitens.

Toxicity of solvents and surfactants to Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) and Dermacentor nitens (Neumann, 1897) (Acari: Ixodidae) larvae.

The objective of this work was to verify the sensitivity of Amblyomma cajennense and Dermacentor nitens larvae to the solvents ethanol, methanol, acetone, xylol and dimethyl sulfoxide (DMSO) and the surfactant Tween 80. The first four solvents were tested at analytical purity while the DMSO and surfactant Tween 80 were tested at a concentration of 1%. The substances tested at analytical purity that caused high mortality were also tested at concentrations of 50%, 25% and 1%. The larval packet test was used, with 10 repetitions for each treatment. A control group was also formed with the same number of repetitions, in which the larvae were only exposed to distilled water. In the first experiment, only xylol was highly toxic at the concentrations tested, causing mortality above 90% for larvae of both species. In the second experiment, xylol at 1% and at 25% showed low toxicity to the A. cajennense and D. nitens larvae, respectively, since the percentage mortality was statistically similar to that of the control group (p>0.05).

Acaricidal activity of hydroethanolic formulations of thymol against Rhipicephalus sanguineus (Acari: Ixodidae) and Dermacentor nitens (Acari: Ixodidae) larvae.

The aim of this study was to assess the acaricidal activity of hydroethanolic formulations of thymol at varying concentrations on Rhipicephalus sanguineus and Dermacentor nitens larvae. The larval packet test was used and the thymol concentrations tested were 2.5, 5.0, 10.0, 15.0 and 20.0 mg/ml. The control group was exposed only to water and ethanol (50/50%) and there were 10 repetitions for each treatment. The mortality was evaluated after 24 h. For the R. sanguineus larvae, the mortality rates were 47.5, 50.2, 96.7, 95.9 and 98.1%, while for D. nitens the rates were 14.1, 75.0, 90.2, 90.3 and 99.5%, at respective thymol concentrations of 2.5, 5.0, 10.0, 15.0 and 20.0 mg/ml. The results indicate that the hydroethanolic formulations of thymol tested have acaricidal activity on R. sanguineus and D. nitens larvae exposed topically, causing mortality greater than 90% 24 h post-treatment starting at the concentration of 10 mg/ml.