Mecanismo de infecção do fungo Metarhizium anisopliae no carrapato Boophilus microplus em condições experimentais The penetration of the fungus Metarhizium anisopliae on Boophilus microplus in experimental conditions

O objetivo do presente trabalho foi avaliar o mecanismo de penetração do fungo Metarhizium anisopliae em Boophilus microplus em condições experimentais. As infecções foram realizadas utilizando amostra de M. anisopliae isolada de carrapatos. A suspensão de conídios do fungo foi preparada utilizando água destilada e espalhante adesivo Tween 80 e foi quantificada na concentração de 10(8) conídios/ml. As fêmeas ingurgitadas foram banhadas nesta suspensão durante cinco minutos e, após este período, foram levadas à câmara climatizada. Três dias após a infecção, as fêmeas foram fixadas utilizando tetróxido de ósmio e glutaraldeído, posteriormente desidratadas em bateria de álcool etílico com acetona. Após a desidratação, esse material foi levado à metalização e ao microscópio eletrônico de varredura, onde se observou a fixação dos conídios na cutícula das fêmeas, havendo germinação e a dilatação da extremidade do tubo germinativo em todas as amostras. Esta dilatação observada foi devido à formação do apressório, apesar de não ser possível observar esta estrutura sobre a cutícula das fêmeas ingurgitadas. O tetróxido de ósmio mostrou ser um excelente fixador para fungos, enquanto o glutaraldeído mostrou ser eficiente na fixação do artrópode. Conclui-se que a forma principal de penetração deste entomopatógeno em B. microplus é através da cutícula. Cabe aqui frisar que esta é a primeira vez que é descrito o mecanismo de penetração de M. anisopliae em carrapatos.<br>The objective of the present communication has been to draw attention to the mode of penetration of the fungus Metarhizium anisopliae through the cuticle of Boophilus microplus under experimental conditions. Samples of M. anisopliae isolated from naturally infected ticks were used to prepare a suspension of 108 conidia/ml in a destilled water with 2% of Tween 80. The engorged females were submergged in this suspension for five minutes, then transferred to 27 &plusmn; 1 &deg;C and > 80% RH in climatized chamber. Three days after the infection, females were fixed in either osmium tetroxide or glutaraldehyde, and dehydrated in alcohol / acetone for SEM. Fixation of conidia in the cuticle, formation of germinative tube and appressoria were seem. It was suggested that the penetration of this entomopathogenic fungus is primarily through the cuticle. The fixation of the fungus for SEM was best accomplished by the osmium tetroxide whereas the glutaraldehyde was more efficient fixative for the tick. It is the first time that the penetration’s mechanism of M. anisopliae is described in ticks

Efeitos de Beauveria bassiana (Bals) Vuill e Metarhizium anisopliae (Metsc) Sorok sobre fêmeas ingurgitadas de Amblyomma cajennense (Fabricius, 1787) em condições de laboratório Effects of Beauveria bassiana (Bals) Vuill and Metarhizium anisopliae (Metsc) Sorok on engorged females of Amblyomma cajennense (Fabricius, 1787) in laboratory conditions

<abstract language="eng">The in vitro susceptibility of Amblyomma cajennense engorged females to some isolated of the fungus Beauveria bassiana and Metarhizium anisopliae was verified and lethal concentrations (LC 50 and LC 90) were calculated. The females were dived in conidia suspensions for five minutes, and kept in climatically controlled chambers BOD under 27&deg; C and 80% relative humidity. Each bioassay had four treatments in concentrations of 10(5),10(6),10(7)e10(8) conidia/ml. A control group was also used. The following characteristics were evaluated: weight and period of oviposition, indexes of reproductive and nutritional efficiency and percentage of microbiological control. A dose dependent negative effect was observed in ticks treated with the suspension. All isolates tested cause a negative effect on in vitro tests of engorged females of A. cajennense, suggesting its potential for microbiological control of tick's species

Ação in vitro dos fungos Beauveria bassiana (Bals) Vuill e Metarhizium anisopliae (Metsch) Sorok sobre ninfas e adultos de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae) In vitro action of the fungi Beauveria bassiana (Bals) Vuill and Metarhizium anisopliae (Metsch) Sorok on ninphs and adults of Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)

O trabalho teve por objetivo verificar a mortalidade in vitro de ninfas e adultos de Amblyomma cajennense frente à ação de isolados dos fungos Beauveria bassiana e Metarhizium anisopliae. Foram avaliados três isolados de M. anisopliae (959, 319 e E9) e dois de B. bassiana (986 e 747). As suspensões de conídios foram preparadas a partir de fungos produzidos em meio de arroz, e cada bioensaio foi constituído de quatro tratamentos nas concentrações 10(5), 10(6), 10(7), 10(8) conídios/ml e um grupo-controle. A análise constou da observação do percentual de mortalidade, 15 dias após o tratamento ou após a ecdise de adultos. Foram observadas diferenças significativas entre os tratamentos quanto à mortalidade para todos os isolados e todos os instares. Nos grupos tratados houve aumento considerável na mortalidade à medida que se aumentava a concentração de conídios na suspensão. Conclui-se que todos os isolados testados causaram mortalidade em testes in vitro sobre esses estádios evolutivos, sugerindo o controle do A. cajennense pela ação desses fungos.<br>This work aimed at the evaluation of the in vitro susceptibility of Amblyomma cajennense nimphs and adults to isolates of Beauveria bassiana and Metarhizium anisopliae fungi. Three isolates of M. anisopliae (959, 319 e E9) and two of B. bassiana (986 e 747) were used. Conidia suspensions were made from fungi grown up in a rice culture medium and each test consisted of four treated groups (10(5), 10(6), 10(7), 10(8) conidia/ ml) plus a control group. Viability of individuals was assessed 15 days after treatment or after adult ecdisis. Significative differences were found for all treatments and for all stages studied. A large reduction in the viability of ticks was observed, and this effect increased as conidia concentration raised. Based upon the results obtained, it can be concluded that all isolates tested presented an in vitro lethality for the biological stages of A. cajennense evaluated, suggesting a potential use in the biological control of this tick species

Ação do fungo Beauveria bassiana (Balsamo) Vuillemin, 1912 sobre a fase parasitária do carrapato Anocentor nitens (Neumann, 1897) Schulze, 1937 (Acari: ixodidae) Action of the fungus Beauveria bassiana (Balsamo) Vuillemin, 1912 on the parasitic phase of the tick Anocentor nitens (Neumann, 1897) Schulze, 1937 (Acari: ixodidae)

A patogenicidade do fungo Beauveria bassiana (isolado 986) foi avaliada no desenvolvimento da fase parasitária do Anocentor nitens, acompanhando o desenvolvimento de larvas não alimentadas até o estágio adulto. Foram utilizados dois tratamentos e um grupo controle com cinco repetições cada. No tratamento 1 (T1), as orelhas dos bovinos foram banhadas com a suspensão fúngica de 10(8) conídios ml-1; no tratamento 2 (T2), as orelhas foram banhadas com água destilada e espalhante adesivo. No grupo controle (C), as orelhas foram banhadas com água destilada. Após 24 horas dos tratamentos, as orelhas de cada animal foram infestadas com 100mg de larvas. Os parâmetros analisados, em cada tratamento, foram: número e peso de fêmeas recuperadas; períodos de pré-postura, postura, incubação, eclosão e longevidade; percentual de eclosão; peso da massa de ovos e da quenógina; índices de eficiência reprodutiva e nutricional. Verificou-se que, no T1 e T2, o número de fêmeas recuperadas, após 35 dias, foi significantemente menor (P<0,05) que no C. O número de fêmeas recuperadas foi 35, 70 e 117 fêmeas, no T1, T2 e C, respectivamente. As fêmeas oriundas das larvas sobreviventes dos grupos tratados não sofreram ação deletéria do entomopatógeno avaliado.<br>This study evaluated the pathogenicity of the fungus Beauveria bassiana (Isolate 986) on the development of the parasitic phase of Anocentor nitens by accompanying the development of unfed larvae until the adult stage. Two treated and one control group were used with five repetitions each. In the first treated group (T1), cattle ears were bathed with a fungal suspension of 10(8) conidia ml-1, second treated group (T2), the ears were bathed with distilled water and Tween 80 1% and the control group (C), the ears were bathed with distilled water. One day after treatment, the ears of each animal were infested with 100 mg of larvae. The parameters analyzed in each treatment were: number and weight of females recovered; periods of pre-oviposition, oviposition, incubation, eclosion and longevity; percentage of eggs hatched; weight of the egg mass and of females after oviposition. We found that in the two treated groups the number of females recovered was significantly lower (P<0.05) than in the control group. 35, 70 and 117 females were recovered from the T1, T2 and C, respectively. The females obtained from the surviving larvae of the treated groups did not suffer any deleterious effects from the entomopathogen studied

Susceptibility of different populations of ticks to entomopathogenic fungi

AbstractThis study aimed to evaluate the in vitro effect of the entomopathogenic fungi Metarhizium anisopliae sensu lato (s.l.) and Beauveria bassiana sensu lato (s.l.) on two distinct populations of Rhipicephalus microplus, from two different experimental farms. Bioassays were performed with engorged females, eggs and larvae. Fungal infection was evaluated based on biological parameters of treated engorged females, percentage of hatch from treated eggs, and percentage of mortality and mean lethal time (LT90) of treated larvae. When the treatments were compared between the two populations, there were significant differences in the following parameters: pre-oviposition period, hatching period and egg production index. Moreover, the results showed that B. bassiana s.l., isolate Bb 986, was more virulent than M. anisopliae s.l., isolate 959, for engorged females, showing a control percentage of 49%. In the bioassay with eggs, the hatching percentage ranged from 3.1% to 49.5% in one population and from 3.4% to 42.7% in the other, with no significant difference between the two populations. In the bioassay with unfed larvae, the mortality percentage ranged from 91.8% to 98.7% in one population and from 71.0% to 94.0% in the other. The LT90 varied from 19.52 to 27.51days in one of the populations and 22.89 to 37.31days in the other. These results suggest that populations of R. microplus show distinct variation in their susceptibility to B. bassiana s.l. and M. anisopliae s.l

Assessment of weight gain and control of parasitism by Rhipicephalus microplus in dairy cattle in the field using organosynthetic parasiticide, phytotherapics and homeopathy

ABSTRACT The goal of this research was to compare the effect of the following products on dairy cattle, parasitized by Rhipicephalus microplus: organosynthetics - Clorfenvinfós and Ivermectina (OG); phytotherapics - Eucalyptus oil (EG) and Neem cake (NG); Homeopathy (HG); The control group (CG) received no treatment. Infestation was by R. microplus (MIC) and weight gain (GP) were evaluated. The study included 60 animals (5 groups) from – Dutch / Zebu, aged between 25-44 months and initial weight between 211-477kg. EG and NG showed MIC 84.9% and 14.0% greater than CG, respectively (P &lt;0.05; P = 0). HG and OG had MIC less than CG 24.4% and 16.9%, respectively (P&lt;0.05; P &lt;0.05). EG, NG and OG obtained GP lower than CG in 7.9%, 8.9% and 8.06% (P &lt;0.05) respectively. The HG GP was 4.9% higher (P &lt;0.05) than CG. This was the first research to prove parasitism control by R. microplus in a field test, using homeopathy. The methodology for choosing Homeopathy for the control of enzooties, developed and tested for the first time in this study, proved to be adequate and efficient, opening the possibility for establishing a new methodology for strategic control of parasitism by R. microplus.</jats:p