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STUDY OF THE EFFECT OF FUNGI BEAUVERIA BASSIANA ON THE ADULTS OF A HARD TICKES BOOPHILUS MICROPLUS

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  • University of Almaarif
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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 108 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.
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Nymph and adult ticks from Ambylomma americanum and Ambylomma maculatum were treated with conidia and blastospores of the entomopathogenic fungi Beauveria bassiana (90517) and Metarhizium anisopliae (20500). Fungal suspensions of conidia harvested from potato dextrose plates containing 108conidia/ml caused greater than 90% mortality in adult A. maculatum but less than 10% mortality in adult A. americanum over a 28 day time course. Similarly, infection with M. anisopliae (108conidia/ml) resulted in 60 and 15% mortality in A. maculatum and A. americanum, respectively. Nymphs of both tick species were more susceptible to fungal infection reaching mortality rates of almost 100% for A. maculatum and over 35% for A. americanum. Scanning electron microscopy of infected ticks showed rapid attachment, germination, and proliferation of fungal spores on A. maculatum cuticles, and to a much lesser extent on A. americanum cuticles. Pentane extracts of A. americanum cuticle hydrocarbons inhibited germination and hyphal growth of B. bassiana conidia, whereas no inhibition was observed using A. maculatum extracts. Significant mortality towards A. americanum was observed (>60%, 28 days) only when the ticks were treated with B. bassiana directly from the growth medium (107blastospores/ml, grown for 3–4 days in Sabouraud dextrose+0.5% yeast extract liquid media). These results indicate tick species display differential susceptibility to the entomopathogenic fungi B. bassiana and M. anisopliae, and that the ability to overcome fungistatic compounds present in the tick epicuticle may determine the likelihood of successful infection and virulence.
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To evaluate the pathogenicity of 4 strains of the entomopathogenic fungi Metarhizium flavoviride var flavoviride and Metarhizium anisopliae var anisopliae in vitro against the bovine tick Boophilus microplus. 300 B. microplus females were used for each series of 5 conidial suspensions of the 4 fungal strains; a control group of 60 females was treated with sterilized water. For each fungal strain, sporulating cultures were used to produce conidial suspensions containing 10(4), 10(5), 10(6), 10(7), and 10(8) conidia/ml. A bioassay was performed by immersing B. microplus females in conidial suspensions; the procedure was repeated with a new group of 60 females for each dilution (a total of 300). A control group was immersed in the same volume of sterilized distilled water. Ticks were maintained at 27 C and 80% relative humidity for 7 days, and the following variables were evaluated: weight before egging, egging weight, percentage eclosion, reproductive efficiency, and percentage control. A low index of eclosion was found in the treated groups, with a progressive decrease in percentage eclosion (control = 100%) with increased conidial concentration. The highest level of effectiveness of biocontrol was detected in groups treated with concentrations of 10(7) and 10(8) conidia/ml. Strains of the entomopathogenic fungi M. anisopliae var anisopliae and M. flavoviride var flavoviride were effective as biological control agents against B. microplus, with M. flavoviride strain CG-291 being the most effective. The pathogenicity of these strains and the development of an application method for this fungus for control of ticks may result in improved animal productivity and a reduction in environmental pollution through the use of fewer chemical insecticides.
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Entomopathogenic fungi are currently being used for the control of several insect pests as alternatives or supplements to chemical insecticides. Improvements in virulence and speed of kill can be achieved by understanding the mechanisms of fungal pathogenesis and genetically modifying targeted genes, thus improving the commercial efficacy of these biocontrol agents. Entomopathogenic fungi, such as Beauveria bassiana, penetrate the insect cuticle utilizing a plethora of hydrolytic enzymes, including chitinases, which are important virulence factors. Two chitinases (Bbchit1 and Bbchit2) have previously been characterized in B. bassiana, neither of which possesses chitin-binding domains. Here we report the construction and characterization of several B. bassiana hybrid chitinases where the chitinase Bbchit1 was fused to chitin-binding domains derived from plant, bacterial, or insect sources. A hybrid chitinase containing the chitin-binding domain (BmChBD) from the silkworm Bombyx mori chitinase fused to Bbchit1 showed the greatest ability to bind to chitin compared to other hybrid chitinases. This hybrid chitinase gene (Bbchit1-BmChBD) was then placed under the control of a fungal constitutive promoter (gpd-Bbchit1-BmChBD) and transformed into B. bassiana. Insect bioassays showed a 23% reduction in time to death in the transformant compared to the wild-type fungus. This transformant also showed greater virulence than another construct (gpd-Bbchit1) with the same constitutive promoter but lacking the chitin-binding domain. We utilized a strategy where genetic components of the host insect can be incorporated into the fungal pathogen in order to increase host cuticle penetration ability.
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Metarhizium anisopliae V275 was efficacious against black vine weevil when used alone or in combination with sublethal doses of imidacloprid or fipronil. Both control strategies were effective in peat, coir, bark and peat blends with composted green waste. M. anisopliae can be applied as drench or premixed into growing media. The former gave marginally better control. Over 98% control was achieved when M. anisopliae was applied with 1/10 dilution (sublethal rates) of fipronil. Marginally lower control was achieved when M. anisopliae was used with 1/100 dilution (sublethal rate) of imidacloprid. However, the latter combination gave better control than imidacloprid used at the recommended rate. More conidia were recovered from larvae exposed to sublethal doses of fipronil than imidacloprid and may explain the higher pest control since mortality is dose-related.
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Seven isolates of the fungus Beauveria bassiana (B.b) were assessed for their lethality against Haemaphysalis qinghaiensis, a prevalent tick species in China. Fourteen days after exposure to the isolates B.bAT1, B.bAT5, and B.bAT7 (at 10(8) conidia mL(-1)), the mortality rate had reached 100%. The results indicated that these three B. bassiana isolates were highly virulent against the engorged female H. qinghaiensis ticks. The present study suggests that B. bassiana has potential for biocontrol applications to eradicate H. qinghaiensis.
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The first experimental vaccination against ticks was carried out 60 years ago. Since then, progress has been slow, although the recent commercial release of a recombinant vaccine against Boophilus microplus is significant. The nature of naturally acquired protective immunity against ticks is poorly understood, particularly in the important, domesticated ruminant hosts. Characterization of the antigens of naturally acquired immunity remains limited, although more has been achieved with 'concealed' antigens. Crucial questions remain about the true impact of tick-induced immunosuppression and the effect of immunity on the transmission of tick-borne diseases, despite some fascinating and important recent results, as discussed here by Peter Willadsen and Frans Jongejan.
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Thirty-one isolates of Metarhizium anisopliae were bioassayed against the cattle tick (Boophilus microplus). More than half of the isolates showed a high degree of virulence to ticks. Radial growth curves for growth between 20 degrees C and 40 degrees C were obtained for all isolates. This information together with information on virulence will be important for the selection of isolates suitable to kill ticks on the surface of cattle. A biopesticide for cattle ticks must kill ticks rapidly at temperatures within the upper end of most isolates' growth curves. It was also found that the time taken to achieve 100% tick mortality in vitro using a virulent isolate could be halved by applying conidia in a 10% oil emulsion. Scanning electron microscopy and light microscopy were used to investigate and compare the germination and penetration of conidia formulated in aqueous and oil formulations. It was found that conidia in both formulations were able to germinate and produce appressoria on the surface of ticks in less than 11h. Marked weakness within 26h, followed by extensive hyphal growth on the cuticle characterised the invasion of ticks by M. anisopliae.
Sqptin Function in yeast model system and pathogenic Fungi
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Norris, H.A. Elewski, B.E. and channoum, M.A. 1999. Optimal growth condition for the determination of the anti fungal suscepitibility of three species of dermatophytes with use of a microdilution method. J.Am. Dermat. 40 (6): 509-513.
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Reis, R.C.S. Frnandes, E.K.K. Bittencourt, V.R.E.P. 2007. Effect of fungal formulation on viability of engorged females of Rhipicephalus sanguineus. Annals of the IX Biennial conference of the society for tropical veterinary medicine.Meridae, Mexico, p 197.