Combination of entomopathogenic nematodes with acaricides or essential oil of Lippia triplinervis against Rhipicephalus microplus (Acari: Ixodidae).

This study aimed to evaluate the combination effect of Heterorhabditis bacteriophora HP88 and H. indica LPP1, with the acaricides deltamethrin, amitraz and chlorfenvinphos, and the essential oil (EO) of Lippia triplinervis, against engorged females of Rhipicephalus microplus. In order to verify the effect of acaricides and EO, the adult immersion test was used, and in the groups treated only with entomopathogenic nematodes (EPNs), 150 infective juveniles were used per female. In the treatments with nematodes in combination with the acaricides or EO, the females were immersed in the solutions (acaricide or EO) and then transferred to Petri dishes for application of the nematodes. The treatment with acaricides resulted in a control percentage lower than 70%, except in the group treated with chlorfenvinphos in the second experiment (84.3%). The control percentage was 73% for L. triplinervis EO, and greater than 90% in all the groups treated with nematodes. For treatments with EPNs combined with the acaricides or EO, the efficacy was greater than 95% (except for deltamethrin + HP88), and reached 100% in the treatment with LPP1 + amitraz. It can be concluded that the EPNs at the concentrations tested were compatible with the acaricides deltamethrin, amitraz and chlorfenvinphos, and with the EO of L. triplinervis. These combinations enhance the effect of these control agents.

Efficacy of entomopathogenic nematodes in insect cadaver formulation against engorged females of Rhipicephalus microplus (Acari: Ixodidae) in semi-field conditions.

The present study evaluated, in the laboratory, the virulence of Heterorhabditis bacteriophora LPP30 against engorged females of Rhipicephalus microplus; in addition, we evaluated, in semi-field tests, the effects of four isolates of Heterorhabditis spp. (i.e., H. bacteriophora HP88, Heterorhabditis baujardi LPP7, Heterorhabditis indica LPP1 and H. bacteriophora LPP30) in insect cadaver formulation against the non-parasitic phase of R. microplus. In the first experiment (in vitro), engorged females were exposed, in Petri dish, to H. bacteriophora LPP30 at 75, 150, 300, 600, 1200 and 2400 nematodes/tick (10 ticks per concentration tested). In the second experiment (semi-field), five engorged females and four cadavers of Galleria mellonella infected with nematodes were placed in pots with soil and grown Brachiaria decumbens; the pots were hold outdoor, exposed to natural environment conditions during the tests. In the third experiment (semi-field), 65 days after the cadavers had been placed in the pots for the second experiment, new engorged females (five per pot) were placed in the pots of the groups treated with H. bacteriophora HP88 or H. baujardi LPP7, to assess their persistence in the soil. In the first test, the percent control was higher than 95% in all treatment groups. In the second experiment, at day 22, the mean mortality rate was 78% in the groups treated with H. bacteriophora LPP30 or H. indica LPP1, and reached 100% and 98% when treated with H. bacteriophora HP88 and H. baujardi LPP7, respectively. In this experiment, the egg-laying inhibition index was higher than 90% in the groups treated with H. bacteriophora HP88 (97.2%) or H. baujardi LPP7 (91.9%). In the third experiment with H. bacteriophora HP88 and H. baujardi LPP7, the egg-laying inhibition index was 59.1% and 43.1%, respectively. We concluded that the isolate LPP30 was highly virulent under laboratory conditions, whereas in semi-field tests, HP88 and LPP7 were the most effective isolates. Moreover, HP88 and LPP7 remained active against engorged females of R. microplus in the soil for 65 days after application of EPN-infected cadavers of G. mellonella.

Evaluation of synergism and development of a formulation with thymol, carvacrol and eugenol for Rhipicephalus microplus control.

The acaricidal activity of combinations of thymol, carvacrol and eugenol was evaluated on larvae and engorged females of the cattle tick Rhipicephalus microplus. The first step assessed the compounds separately, in concentrations of 3.125, 6.25, 12.5 and 25 mg/mL. Then tests were performed with the compounds combined in the ratio of 1:1 at concentrations of 3.125 and 6.25 mg/mL, along with the control group treated with the solvent (3% DMSO). In the second step, combinations were tested incorporated in a formulation at the concentration de 6.25 mg/mL, using the larval packet and adult immersion tests. The associations carvacrol + thymol (3.125 mg/mL), carvacrol + eugenol and thymol + eugenol (6.25 mg/mL) presented synergism, while the other associations had an additive effect. In the experiments with formulation, all combinations caused 100% larval mortality, but the efficacy was under 15% against engorged females. Therefore, the combinations of thymol + carvacrol (3.125 mg/mL) as well as carvacrol + eugenol and eugenol + thymol (6.25 mg/mL) had a synergistic effect on engorged females, but when incorporated in the formulation, the acaricide activity was strong against larvae but weak against engorged females.

Lab-on-a-chip and SDS-PAGE analysis of hemolymph protein profile from Rhipicephalus microplus (Acari: Ixodidae) infected with entomopathogenic nematode and fungus.

In the present study, lab-on-a-chip electrophoresis (LoaC) was suggested as an alternative method to the conventional polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE) to analyze raw cell-free tick hemolymph. Rhipicephalus microplus females were exposed to the entomopathogenic fungus Metarhizium anisopliae senso latu IBCB 116 strain and/or to the entomopathogenic nematode Heterorhabditis indica LPP1 strain. Hemolymph from not exposed or exposed ticks was collected 16 and 24 h after exposure and analyze by SDS-PAGE or LoaC. SDS-PAGE yielded 15 bands and LoaC electrophoresis 17 bands. Despite the differences in the number of bands, when the hemolymph protein profiles of exposed or unexposed ticks were compared in the same method, no suppressing or additional bands were detected among the treatments regardless the method (i.e., SDS-PAGE or chip electrophoresis using the Protein 230 Kit®). The potential of LoaC electrophoresis to detect protein bands from tick hemolymph was considered more efficient in comparison to the detection obtained using the traditional SDS-PAGE method, especially when it comes to protein subunits heavier than 100 KDa. LoaC electrophoresis provided a very good reproducibility, and is much faster than the conventional SDS-PAGE method, which requires several hours for one analysis. Despite both methods can be used to analyze tick hemolymph composition, LoaC was considered more suitable for cell-free hemolymph protein separation and detection. LoaC hemolymph band percent data reported changes in key proteins (i.e., HeLp and vitellogenin) exceptionally important for tick embryogenesis. This study reported, for the first time, tick hemolymph protein profile using LoaC.

Streptococcus mutans Can Modulate Biofilm Formation and Attenuate the Virulence of Candida albicans.

Streptococcus mutans and Candida albicans are found together in the oral biofilms on dental surfaces, but little is known about the ecological interactions between these species. Here, we studied the effects of S. mutans UA159 on the growth and pathogencity of C. albicans. Initially, the effects of S. mutans on the biofilm formation and morphogenesis of C. albicans were tested in vitro. Next, we investigate the influence of S. mutans on pathogenicity of C. albicans using in vivo host models, in which the experimental candidiasis was induced in G. mellonella larvae and analyzed by survival curves, C. albicans count in hemolymph, and quantification of hyphae in the host tissues. In all the tests, we evaluated the direct effects of S. mutans cells, as well as the indirect effects of the subproducts secreted by this microorganism using a bacterial culture filtrate. The in vitro analysis showed that S. mutans cells favored biofilm formation by C. albicans. However, a reduction in biofilm viable cells and inhibition of hyphal growth was observed when C. albicans was in contact with the S. mutans culture filtrate. In the in vivo study, injection of S. mutans cells or S. mutans culture filtrate into G. mellonella larvae infected with C. albicans increased the survival of these animals. Furthermore, a reduction in hyphal formation was observed in larval tissues when C. albicans was associated with S. mutans culture filtrate. These findings suggest that S. mutans can secrete subproducts capable to inhibit the biofilm formation, morphogenesis and pathogenicity of C. albicans, attenuating the experimental candidiasis in G. mellonella model.

Competitive Interactions between C. albicans, C. glabrata and C. krusei during Biofilm Formation and Development of Experimental Candidiasis.

In this study, we evaluated the interactions between Candida albicans, Candida krusei and Candida glabrata in mixed infections. Initially, these interactions were studied in biofilms formed in vitro. CFU/mL values of C. albicans were lower in mixed biofilms when compared to the single biofilms, verifying 77% and 89% of C. albicans reduction when this species was associated with C. glabrata and C. krusei, respectively. After that, we expanded this study for in vivo host models of experimental candidiasis. G. mellonella larvae were inoculated with monotypic and heterotypic Candida suspensions for analysis of survival rate and quantification of fungal cells in the haemolymph. In the groups with single infections, 100% of the larvae died within 18 h after infection with C. albicans. However, interaction groups achieved 100% mortality after 72 h of infection by C. albicans-C. glabrata and 96 h of infection by C. albicans-C. krusei. C. albicans CFU/mL values from larvae hemolymph were lower in the interacting groups compared with the monoespecies group after 12 h of infection. In addition, immunosuppressed mice were also inoculated with monotypic and heterotypic microbial suspensions to induce oral candidiasis. C. albicans CFU/mL values recovered from oral cavity of mice were higher in the group with single infection by C. albicans than the groups with mixed infections by C. albicans-C. glabrata and C. albicans-C. krusei. Moreover, the group with single infection by C. albicans had a higher degree of hyphae and epithelial changes in the tongue dorsum than the groups with mixed infections. We concluded that single infections by C. albicans were more harmful for animal models than mixed infections with non-albicans species, suggesting that C. albicans establish competitive interactions with C. krusei and C. glabrata during biofilm formation and development of experimental candidiasis.

Acaricidal properties of the formulations based on essential oils from Cymbopogon winterianus and Syzygium aromaticum plants.

The cattle tick, Rhipicephalus (Boophilus) microplus, has caused serious harm to livestock raising in Brazil, considering the costs of controlling it, loss of revenue due to smaller production of milk and meat, and damage to leather, in addition to transmitting diseases. The use of medicinal plants is considered an alternative to the recurring resistance to chemicals. Due to the need for efficient alternatives with less environmental impact, this study aimed to develop contact formulations with essential oils from the Java citronella (Cymbopogon winterianus) and clove (Syzygium aromaticum) plants and to assess in vitro the effects in different stages of the tick cycle. In the present study, concentrations from 0.5-15.0% of the essential oils incorporated in the formulations were used. The ticks from different geographical areas were treated with those formulations, and their effects on the production levels of eggs, on the larvae hatching, and their efficiency on ticks were assessed. The obtained results were compared with other commercial acaricidal products. After the 20th day of treatment, the formulations with citronella essential oil had 2.09-55.51% efficiency, depending on the concentration of the oil incorporated. The efficiency of the treatment with formulations containing clove essential oil was higher, from 92.47-100%. The results showed the acaricidal effects of the formulations tested when compared to commercial chemical products. In vivo studies should be performed in order to assess the efficiency of those formulations in the fields, aiming to use these products as an alternative for controlling cattle ticks.

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)

Entomopathogenic nematodes associated with essential oil of Lippia sidoides for control of Rhipicephalus microplus (Acari: Ixodidae).

In vitro effect of the association of the entomopathogenic nematodes Heterorhabditis bacteriophora HP88 and Heterorhabditis indica LPP1 with the essential oil of Lippia sidoides on engorged females of Rhipicephalus microplus was assessed. Engorged females with homogeneous weights (p >0.05) were divided into six groups of ten ticks each (each female = an experimental unit). In the treated groups, the ticks were exposed to the oil (40 µl/ml=4%) and nematodes (300 entomopathogenic nematodes (EPNs/tick) separately and also to the nematodes together with the oil, while in the control group, the females were immersed in Tween 80 (3%). All the treatments caused a significant reduction in the egg mass weight (p <0.05). The hatching percentage of larvae was reduced (p <0.05) in the groups treated with H. bacteriophora HP88, H. indica LPP1, and H. bacteriophora HP88 associated with the oil, but in the group treated only with the oil, the hatching of percentage of larvae rate was similar to that of the control group (p >0.05). The percentage of control in the group exposed only to the oil was 50.7%, while in the other treated groups, the percentage of control was greater than 95 %, reaching 100 %in the group treated with H. indica LPP1 associated with the oil. The results of the L. sidoides essential oil in the concentrations tested in association with both H. bacteriophora HP88 and H. indica LPP1 indicate that these combinations can be used to control R. microplus.

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.

Biological parameters of cattle ticks fed on rabbits.

With the objective of encouraging the use of rabbits as alternative hosts for the cattle tick, four rabbits were infested on the ears and back. From the second day of infestation the developmental stages were observed. e duration of larval engorgement and changes were estimated between six and eight days in the region of the back and between 've and seven days in the pinna. e nymphal engorgement and changes occurred at approximately 10.80 ± 2.65 days in the dorsal and 11.00 ± 2.52 days in the ear, with the engorgement of adults and copulation occurring at 7.03 ± 2.45 days, on the dorsal region and 8.55 ± 1.82 days in the region of the pinna. e parasitic period ranged from 21 to 29 days (back) and from 23 to 30 days (pinna). e engorged females of the back and ear weighed on average 34.43 ± 18.73 and 36.30 ± 18.10 mg, respectively. e nutritional and reproductive efficiency indexes were 17.38 ± 14.27 and 26.85 ± 17.13% (back) and 17.42 ± 12.22% and 30.70 ± 19.80% (pinna). Although not appropriate to maintain successive generations of Rhipicephalus (Boophilus) microplus, rabbits can be used for different stages of engorgement of the ixodid, allowing experimental studies.

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).

Biological parameters of cattle ticks fed on rabbits / Parâmetros biológicos de carrapatos bovinos alimentados em coelhos

With the objective of encouraging the use of rabbits as alternative hosts for the cattle tick, four rabbits were infested on the ears and back. From the second day of infestation the developmental stages were observed. e duration of larval engorgement and changes were estimated between six and eight days in the region of the back and between 've and seven days in the pinna. e nymphal engorgement and changes occurred at approximately 10.80 ± 2.65 days in the dorsal and 11.00 ± 2.52 days in the ear, with the engorgement of adults and copulation occurring at 7.03 ± 2.45 days, on the dorsal region and 8.55 ± 1.82 days in the region of the pinna. e parasitic period ranged from 21 to 29 days (back) and from 23 to 30 days (pinna). e engorged females of the back and ear weighed on average 34.43 ± 18.73 and 36.30 ± 18.10 mg, respectively. e nutritional and reproductive e/ciency indexes were 17.38 ± 14.27 and 26.85 ± 17.13% (back) and 17.42 ± 12.22% and 30.70 ± 19.80% (pinna). Although not appropriate to maintain successive generations of Rhipicephalus (Boophilus) microplus, rabbits can be used for di:erent stages of engorgement of the ixodid, allowing experimental studies.

Com objetivo de viabilizar o uso de coelhos como hospedeiros alternativos do carrapato bovino, foram infestados quatro coelhos nas orelhas e no dorso. A partir do segundo dia da infestação, foram observados os estágios de desenvolvimento dos carrapatos. A duração do ingurgitamento e muda larval foi estimada entre seis e oito dias na região do dorso e entre cinco e sete dias no pavilhão auricular. O ingurgitamento e muda ninfal ocorreram em aproximadamente, 10,80 ± 2,65 dias na região dorsal e 11,00 ± 2,52 dias no pavilhão auricular; e o ingurgitamento e cópula dos adultos ocorreu em 7,03 ± 2,45 dias, na região do dorso e em 8,55 ± 1,82 dias na região do pavilhão auricular . O período parasitário variou de 21 a 29 dias (dorso) e de 23 a 30 dias (pavilhão auricular). As fêmeas ingurgitadas do dorso e orelha pesaram, em média, 34,43 ± 18,73 e 36,30 ± 18,10 mg, respectivamente. Os índices de e'ciência reprodutiva e nutricional foram de 17,38 ± 14,27 e 26,85 ± 17,13% (região do dorso) e de 17,42 ± 12,22% e 30,70 ± 19,80% (pavilhão auricular). Embora não sejam adequados para manter sucessivas gerações de Rhipicephalus (Boophilus) microplus, os coelhos podem ser empregados para o ingurgitamento dos diferentes estádios deste ixodídeo, viabilizando estudos experimentais.

Heterorhabditis bacteriophora (Rhabditida: Heterorhabditidae) HP88 for biological control of Rhipicephalus microplus (Acari: Ixodidae): the effect of different exposure times of engorged females to the nematodes.

The aim of this study was to assess the influence of different exposure times of engorged female the Rhipicephalus microplus to infective juveniles of Heterorhabditis bacteriophora isolate HP88. The engorged females were divided into seven groups (six treatments and one control) of 20 ticks each with statistically similar average weights (p>0.05) and exposed to concentrations of 300 nematodes/tick for periods of 3, 6, 12, 24, 48 and 72 h. The following biological parameters were monitored: pre-oviposition period; egg mass weight; hatching percentage; and efficacy of treatment. There was no influence of the exposure time on the pre-oviposition period (p>0.05), while there were significant differences (p<0.05) between the groups exposed for 24, 48 and 72 h and the control group with respect to the egg mass weight, and 24h for hatching percentage. Treatment efficacy reached 100% after exposure for 48 and 72 h. These results demonstrate that infective juveniles of H. bacteriophora HP88, under laboratory conditions, interfere in the majority of the biological parameters of the non-parasitic phase of engorged R. microplus females when the exposure time is greater than or equal to 24h.

Perceptions and attitudes among milk producers in Minas Gerais regarding cattle tick biology and control.

This study evaluates milk producers' knowledge regarding cattle ticks and practices for controlling them. Ninety-three dairymen in Minas Gerais were interviewed. These producers had no information regarding acaricide efficiency tests. To analyze the information, open responses were categorized through "content analysis", and descriptive analysis consisting of extracting the profile highlighted by the highest frequencies. The association between schooling level and knowledge was tested by means of chi-square trend tests. It was observed that 92.3% had no knowledge of the nonparasitic period. For 96.4%, what determined the time to apply treatment was the degree of tick infestation; 93.3% used spray guns to apply the acaricide. In seeking to cross-correlate the biological and control variables with education, cooperative action, length of experience and herd size, it was found that there was a linear association between schooling level and implementation of acaricide solution preparation. The other factors didn't show any significant association. These data demonstrated the need to instruct the producers in relation to the biology and control of Rhipicephalus (Boophilus) microplus. It was concluded that the majority of milk producers were unaware of cattle tick biology and the factors that influence choosing an acaricide, which makes it difficult to implement strategic control.

Perceptions and attitudes among milk producers in Minas Gerais regarding cattle tick biology and control / Percepções e atitudes entre produtores de leite em Minas Gerais relacionado a biologia e controle de carrapatos em bovinos

This study evaluates milk producers' knowledge regarding cattle ticks and practices for controlling them. Ninety-three dairymen in Minas Gerais were interviewed. These producers had no information regarding acaricide efficiency tests. To analyze the information, open responses were categorized through "content analysis", and descriptive analysis consisting of extracting the profile highlighted by the highest frequencies. The association between schooling level and knowledge was tested by means of chi-square trend tests. It was observed that 92.3 percent had no knowledge of the non-parasitic period. For 96.4 percent, what determined the time to apply treatment was the degree of tick infestation; 93.3 percent used spray guns to apply the acaricide. In seeking to cross-correlate the biological and control variables with education, cooperative action, length of experience and herd size, it was found that there was a linear association between schooling level and implementation of acaricide solution preparation. The other factors didn't show any significant association. These data demonstrated the need to instruct the producers in relation to the biology and control of Rhipicephalus (Boophilus) microplus. It was concluded that the majority of milk producers were unaware of cattle tick biology and the factors that influence choosing an acaricide, which makes it difficult to implement strategic control.

Este estudo teve como objetivo avaliar o conhecimento dos produtores de leite sobre o carrapato dos bovinos e seu controle. Foram entrevistados 93 produtores de leite de Minas Gerais. Estes produtores não tinham informação sobre testes de eficiência de carrapaticidas e controle de carrapatos. Foi testada associação entre a escolaridade e as práticas e conhecimento sobre os carrapatos e constatou-se que 92,3 por cento dos produtores nada sabiam sobre o período não-parasitário. Para 96,4 por cento, o que determinava o momento do tratamento era o grau de infestação de carrapatos; e 93,3 por cento utilizavam bomba-costal para aplicação do carrapaticida. Ao buscar o cruzamento entre as variáveis de biologia e controle com a escolaridade, cooperativismo, tempo de experiência e tamanho do rebanho, verificou-se que a associação linear entre a escolaridade e a efetivação da pré-diluição no preparo de banhos carrapaticidas. Esses dados demonstraram a necessidade de instrução dos produtores em relação à biologia e ao controle do Rhipicephalus (Boophilus) microplus. Conclui-se que a maioria dos produtores de leite de MG desconhece a biologia dos carrapatos e os fatores que influenciam as escolhas do carrapaticida, o que dificulta a implementação do controle estratégico e que a escolaridade não está relacionada a um controle mais eficiente dos carrapatos nas propriedades.

Strategic control of cattle ticks: milk producers' perceptions.

In order to evaluate milk producers' knowledge about tests on tick sensitivity to acaricides, efficiency tests and strategic control, questionnaires were delivered to 670 producers. From these, 163 responses were received. These producers had sent ingurgitated female ticks to be subjected to tests on tick sensitivity to acaricides at Embrapa between 2001 and 2005. Most of the completed questionnaires came from the State of Minas Gerais, Brazil. After descriptive analysis and correlation of variables, it was found that 48.5% of the properties owned herds of up to 100 head of cattle and 57.3% of the producers belonged to a cooperative. Among the respondents, 157 (98.7%) applied the acaricide indicated by the efficiency test, and 144 (92.9%) said that they applied strategic control but incorrectly. There was improvement in some control stages. However, only 12 (7%) showed comprehension of all the stages of strategic control. It was concluded that there is a need for continuing assistance, so as to achieve effective improvement in controlling Rhipicephalus (Boophilus) microplus.

Strategic control of cattle ticks: milk producers' perceptions / Controle estratégico do carrapato dos bovinos: percepção dos produtores de leite

In order to evaluate milk producers' knowledge about tests on tick sensitivity to acaricides, efficiency tests and strategic control, questionnaires were delivered to 670 producers. From these, 163 responses were received. These producers had sent ingurgitated female ticks to be subjected to tests on tick sensitivity to acaricides at Embrapa between 2001 and 2005. Most of the completed questionnaires came from the State of Minas Gerais, Brazil. After descriptive analysis and correlation of variables, it was found that 48.5 percent of the properties owned herds of up to 100 head of cattle and 57.3 percent of the producers belonged to a cooperative. Among the respondents, 157 (98.7 percent) applied the acaricide indicated by the efficiency test, and 144 (92.9 percent) said that they applied strategic control but incorrectly. There was improvement in some control stages. However, only 12 (7 percent) showed comprehension of all the stages of strategic control. It was concluded that there is a need for continuing assistance, so as to achieve effective improvement in controlling Rhipicephalus (Boophilus) microplus.

Objetivando avaliar o conhecimento dos produtores de leite que usufruíram do teste de sensibilidade dos carrapatos aos carrapaticidas - teste de eficácia e controle estratégico - foram enviados questionários a 670 produtores. Desses, 163 foram respondidos. Esses produtores representam aqueles que enviaram fêmeas ingurgitadas para proceder aos testes de sensibilidade dos carrapatos aos carrapaticidas pela Embrapa, de 2001 a 2005. Dos respondidos, a maioria localiza-se no Estado de Minas Gerais. Após análise descritiva e cruzamento de variáveis, constatou-se que 48,5 por cento das propriedades possuem rebanho até 100 cabeças de gado; 57,3 por cento produtores pertencem a uma cooperativa. Um total de 157 (98,7 por cento) entrevistados aplicou o acaricida indicado pelo teste de eficácia, e 144 (92,9 por cento) respondentes afirmaram ter seguido o controle estratégico, porém, não de forma correta, havendo, entretanto, melhora em algumas etapas do controle. No entanto, apenas 12 (7 por cento) demonstraram compreender todas as fases do controle estratégico, apesar de terem recebido informações técnicas de como proceder com o controle. Conclui-se que há necessidade de assistência continuada para melhoria efetiva do controle de Rhipicephalus (Boophilus) microplus .

Evaluation in vitro of the infection times of engorged females of Rhipicephalus (Boophilus) microplus by the entomopathogenic nematode Steinernema glaseri CCA strain / Avaliação in vitro dos tempos de infecção de fêmeas ingurgitadas de Rhipicephalus (Boophilus) microplus pelo nematoide entomopatogênico Steinernema glaseri estirpe CCA

Studies have shown that ticks are susceptible to infection by entomopathogenic nematodes. These studies indicate different susceptibilities of ticks to infection by these fungi, depending on the tick species, development phase, entomopathogenic nematodes species and strains and the time the ticks are exposed to them. Usually this period ranges from 24 to 72 hours. The aim of this study was to evaluate the infection times in vitro of engorged Rhipicephalus (Boophilus) microplus females by the entomopathogenic nematodes Steinernema glaseri CCA strain, by analysis of the ticks' biological parameters. The results show that a 2-hour exposure time was sufficient for the engorged R. microplus females to be infected by S. glaseri CCA, but that a minimum exposure time of 24 hours was necessary to generate treatment efficacy above 90 percent.

Os carrapatos são susceptíveis à infecção por nematoides entomopatogênicos. Essa susceptibilidade diverge quanto às espécies de carrapato estudadas, à fase evolutiva, às espécies e estirpes dos nematoides e ao tempo ao qual os carrapatos ficam expostos a estes. O presente trabalho teve como objetivo avaliar os tempos de infecção in vitro de fêmeas ingurgitadas de Rhipicephalus (Boophilus) microplus pelo nematoide entomopatogênico Steinernema glaseri estirpe CCA, pela análise dos parâmetros biológicos do carrapato. Os resultados obtidos demonstraram que um período de duas horas de exposição foi suficiente para que fêmeas ingurgitadas de R. microplus fossem infectadas por S. glaseri CCA e que um período de exposição mínimo de 24h foi necessário para que houvesse infecção de fêmeas ingurgitadas de R. microplus por S. glaseri estirpe CCA, capaz de gerar, in vitro, eficácia no tratamento superior a 90 por cento.