The limpet transcription factors of Triatoma infestans regulate the response to fungal infection and modulate the expression pattern of defensin genes

As part of the innate humoral response to microbial attack, insects activate the expression of antimicrobial peptides (AMP). Understanding the regulatory mechanisms of this response in the Chagas disease vector Triatoma infestans is important since biological control strategies against pyrethroid-resistant insect populations were recently addressed by using the entomopathogenic fungus Beauveria bassiana. By bioinformatics, gene expression, and silencing techniques in T. infestans nymphs, we achieved sequence and functional characterization of two variants of the limpet transcription factor (Tilimpet) and studied their role as regulators of the AMP expression, particularly defensins, in fungus-infected insects. We found that Tilimpet variants may act differentially since they have divergent sequences and different relative expression ratios, suggesting that Tilimpet-2 could be the main regulator of the higher expressed defensins and Tilimpet-1 might play a complementary or more general role. Also, the six defensins (Tidef-1 to Tidef-6) exhibited different expression levels in fungus-infected nymphs, consistent with their phylogenetic clustering. This study aims to contribute to a better understanding of T. infestans immune response in which limpet is involved, after challenge by B. bassiana infection.Fil: Mannino, Maria Constanza. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Santos Da Paixao, Flavia Regina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Pedrini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentin

Alkane-grown Beauveria bassiana produce mycelial pellets displaying peroxisome proliferation, oxidative stress, and cell surface alterations

The entomopathogenic fungus Beauveria bassiana is able to grow on insect cuticle18 hydrocarbons as the sole carbon source, inducing several enzymes involved in alkane19 assimilation and concomitantly increasing virulence against insect hosts. In this study, we20 describe some physiological and molecular processes implicated in growth, nutritional21 stress response, and cellular alterations found in alkane-grown fungi. The fungal cytology22 was investigated using light and transmission electron microscopy (TEM) while the surface23 topography was examined using atomic force microscopy (AFM). Fungal hydrophobicity24 was also measured on the cell surface. Additionally, the expression pattern of several genes25 associated with oxidative stress, peroxisome biogenesis, and hydrophobicity were analysed26 by qPCR. We found a novel type of growth in alkane-cultured B. bassiana similar to27 mycelial pellets described in other alkane-free fungi, which were able to germinate and28 produce viable conidia in media without a carbon source and to be pathogenic against29 larvae of the beetles Tenebrio molitor and Tribolium castaneum. Optical microscopy and30 TEM showed that pellets were formed by hyphae cumulates with high peroxidase activity,31 exhibiting peroxisome proliferation and an apparent surface thickening. Alkane-grown32 conidia appeared to be more hydrophobic and cell surfaces displayed different topography33 than glucose-grown cells, as it was observed by AFM. We also found a significant34 induction in several genes encoding for peroxins, catalases, superoxide dismutases, and35 hydrophobins. These results show that both morphological and metabolic changes are36 triggered in mycelial pellets derived from alkane-grown B. bassiana.Fil: Huarte Bonnet, Carla. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Santos Da Paixao, Flavia Regina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Ponce, Juan C. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Santana, Marianela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; ArgentinaFil: Prieto, Eduardo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas; ArgentinaFil: Pedrini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata ; Argentin

The entomopathogenic fungus Beauveria bassiana produces microsclerotia-like pellets mediated by oxidative stress and peroxisome biogenesis

Several filamentous fungi are known to produce macroscopic pigmented hyphal aggregates named sclerotia. In recent years, some entomopathogenic fungi were reported to produce small sclerotia termed ‘microsclerotia’, becoming new potential propagules for biocontrol strategies. In this study, we described the production of microsclerotia-like pellets by the entomopathogenic fungus Beauveria bassiana. The carbon: nitrogen ratio equal to or higher than 12.5:1 amended with Fe2+ induced the germination of conidia, producing hyphal aggregate that formed sclerotial structures in submerged liquid cultures. These aggregates were able to tolerate desiccation as they germinated and subsequently produced viable conidia. Conidia derived from microsclerotial aggregates formulated with diatomaceous earth effectively kill Tribolium castaneum larvae. Optical and transmission microscopical imaging, qPCR and spectrophotometric analysis revealed that an oxidative stress scenario is involved in conidial differentiation into microsclerotia-like pellets, inducing fungal antioxidant response with high peroxidase activity – mainly detected in peroxisomes and mitochondria – and progress with active peroxisome proliferation. The results provide clues about B. bassiana microsclerotial differentiation and indicate that these pigmented aggregates are promising propagules for production, formulation and potentially application in the control of soil-inhabiting arthropod pests.Fil: Huarte Bonnet, Carla. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Santos Da Paixao, Flavia Regina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Mascarin, Gabriel M.. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; BrasilFil: Santana, Marianela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Fernandes, Éverton. Universidade Federal de Goiás; BrasilFil: Pedrini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentin

Efficacy of Metarhizium anisopliae conidia in oil-in-water emulsion against the tick Rhipicephalus microplus under heat and dry conditions

Environmental stress conditions compromise the effectiveness of microorganisms for use in biological control. We investigated the thermotolerance of seven Metarhizium isolates from Central Brazil to 32 °C, and the thermotolerance of conidia of Metarhizium anisopliae s.str. IP 119 (Metchnikoff) (Hypocreales: Clavicipitaceae) and M. robertsii formulated in oil-in-water emulsion to 45 °C. We also evaluated the efficacy of oil-formulated conidia of M. anisopliaes.str. IP 119 against the tick Rhipicephalus microplus (Canestrini) (Acari: Ixodidae) under stress conditions (32 °C; 75% RH). The conidial germination on tick’s cuticle was assessed by Scanning Electron Microscopy. Conidia of M. anisopliaes.str. IP 119 formulated in oil-in-water emulsion caused significant control (59.4%) of R. microplus, even when treated ticks were incubated under stress (32 °C; 75% RH). Metarhizium spp. incubated at 32 °C had both decreased radial growth and conidial production. Conidia of IP 119 formulated in oil-in-water emulsion were more tolerant to 45 °C than conidia suspended in water. An oil-in-water emulsion protected M. anisopliaes.str. conidia against heat and dry conditions and may enhance its efficacy to control ticks by improving the tolerance of fungi against adverse environmental factors.Fil: Muniz, Elen R.. Universidade Federal de Goiás; BrasilFil: Santos Da Paixao, Flavia Regina. Universidade Federal de Goiás; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Barreto, Lucas P.. Universidade Federal de Goiás; BrasilFil: Luz, Christian. Universidade Federal de Goiás; BrasilFil: Arruda, Walquíria. Universidade Federal de Goiás; BrasilFil: Angelo, Isabele C.. Universidade Federal Rural do Rio de Janeiro; BrasilFil: Fernandes, Éverton. Universidade Federal de Goiás; Brasi

Increased heat tolerance afforded by oil-based conidial formulations of Metarhizium anisopliae and Metarhizium robertsii

The thermotolerance of oil-based conidial formulations of Metarhizium anisopliae s.l. (IP 46) and Metarhizium robertsii (ARSEF 2575) were investigated. Conidia of IP 46 or ARSEF 2575 were suspended in different adjuvants and exposed to 45 ± 0.2°C for 4, 6, 8 or 24 h; their viability was then assessed after 48 h incubation at 27 ± 1°C. Conidia heated in pure mineral or vegetable oil exhibited mean relative viability exceeding 70% after 8 h of heat exposure, whereas low germination (≤20%) was observed when conidia were heated in water (Tween 80® 0.01%), carboxymethyl cellulose gel or emulsifiable oils (Graxol® or Assist®) and exposed to heat for 6 or 8 h. In addition, conidia of IP 46 suspended in either pure mineral or canola oil and exposed to heat for 48 h had moderate viability, 57% or 41%, respectively. Unstable oil-in-water emulsions showed a higher percentage of conidia incorporated into oil micellae, while the stable emulsions had higher percentage of conidia outside the oil micellae. The thermotolerance of conidia formulated in stable emulsions, however, did not differ from that of conidia formulated in unstable emulsions. The present study highlights possibilities to alleviate the deleterious effects of heat stress towards Metarhizium spp. conidia applied for controlling arthropod pests and vectors through oil-based formulations.Fil: Santos Da Paixao, Flavia Regina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata ; Argentina. Universidade Federal de Goiás; BrasilFil: Muniz, E.R.. Universidade Federal de Goiás; BrasilFil: Barreto, L. P.. Universidade Federal de Goiás; BrasilFil: Bernardo, C. C.. Universidade Federal de Goiás; BrasilFil: Mascarin, G. M.. Universidade Federal de Goiás; BrasilFil: Luz, C.. Universidade Federal de Goiás; BrasilFil: Fernandes, Éverton. Universidade Federal de Goiás; Brasi

Pathogenicity of microsclerotia from Metarhizium robertsii against Aedes aegypti larvae and antimicrobial peptides expression by mosquitoes during fungal-host interaction

Aedes aegypti is a vector of various disease-causing arboviruses. Chemical insecticide-based methods for mosquito control have increased resistance in different parts of the world. Thus, alternative control agents such as the entomopathogenic fungi are excellent candidates to control mosquitoes as part of an ecofriendly strategy. There is evidence of the potential of entomopathogenic fungal conidia and blastospores for biological control of eggs, larval and adult stages, as well as the pathogenicity of fungal microsclerotia against adults and eggs. However, there are no studies on the pathogenicity of microsclerotia against either aquatic insects or insects that develop part of their life cycle in the water, such as the A. aegypti larvae. In this study, we assayed the production of microsclerotia and their pathogenicity against A. aegypti larvae of two isolates of Metarhizium robertsii, i.e., CEP 423 isolated in La Plata, Argentina, and the model ARSEF 2575. Both isolates significantly reduced the survival of A. aegypti exposed to their microsclerotia. The fungus-larva interaction resulted in a delayed response in the host. This was evidenced by the expression of some humoral immune system genes such as defensins and cecropin on the 9th day post-infection when the fungal infection was consolidated as a successful process that culminates in larvae mortality. In conclusion, M. robertsii microsclerotia are promising propagules to be applied as biological control agents against mosquitoes since they produce pathogenic conidia against A. aegypti larvae.Fil: Santos Da Paixao, Flavia Regina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Falvo, Marianel Lucía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Huarte Bonnet, Carla. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Santana, Marianela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Garcia, Juan Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Fernandes, Éverton. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; BrasilFil: Pedrini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentin

Heat-stressed Metarhizium anisopliae: viability (in vitro) and virulence (in vivo) assessments against the tick Rhipicephalus sanguineus

The current study investigated the thermotolerance of Metarhizium anisopliae s.l. conidia from the commercial products Metarril® SP Organic and Metarril® WP. The efficacy of these M. anisopliae formulations against the tick Rhipicephalus sanguineus s.l. was studied in laboratory under optimum or heat-stress conditions. The products were prepared in water [Tween® 80, 0.01 % (v/v)] or pure mineral oil. Conidia from Metarril® SP Organic suspended in water presented markedly delayed germination after heating to constant 40 °C (for 2, 4, or 6 h) compared to conidia suspended in mineral oil. Metarril® SP Organic suspended in oil and exposed to daily cycles of heat-stress (40 °C for 4 h and 25 °C for 19 h for 5 consecutive days) presented relative germination of conidia ranging from 92.8 to 87.2 % from day 1 to day 5, respectively. Conversely, germination of conidia prepared in water ranged from 79.3 to 39.1 % from day 1 to day 5, respectively. Culturability of Metarril® WP decreased from 96 % when conidia were cultured for 30 min prior to heat exposure (40 °C for 4 h) to 9 % when conidia were cultured for 8 h. Tick percent control was distinctly higher when engorged females were treated with oil suspensions rather than water suspensions, even when treated ticks were exposed to heat-stress regimen. Oil-based applications protected fungal conidia against heat-stress. Although Metarril® is not registered for tick control, it may be useful for controlling R. sanguineus, especially if it is prepared in mineral oil.Fil: Alves, Fabrício M.. Universidade Federal de Goiás; BrasilFil: Bernardo, Cíntia C.. Universidade Federal de Goiás; BrasilFil: Santos Da Paixao, Flavia Regina. Universidade Federal de Goiás; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Barreto, Lucas P.. Universidade Federal de Goiás; BrasilFil: Luz, Wolf Christian. Universidade Federal de Goiás; BrasilFil: Humber, Richard A.. Universidade Federal de Goiás; BrasilFil: Fernandes, Éverton. Universidade Federal de Goiás; Brasi

Metarhizium robertsii and M. acridum conidia produced on riboflavin-supplemented medium have increased UV-A tolerance and upregulated photoprotection and photoreactivation genes

The aim of this study was to evaluate the effect of riboflavin supplementation of culture medium on conidial UV-A tolerance of M. acridum (Driver & Milner) (Hypocreales: Clavicipitaceae) and M. robertsii (Bischoff, Rehner & Humber) (Hypocreales: Clavicipitaceae). These fungi were produced on culture medium supplemented, or not supplemented, with riboflavin. Relative germination and expression patterns of some photoprotection-related genes were evaluated after irradiating with artificial UV-A, or with filtered solar radiation (> 320 nm; UV-A and visible radiation). M. acridum conidia harvested from riboflavin-supplemented culture medium demonstrated enhanced UV-A tolerance when irradiated with artificial UV-A. Nevertheless, relative germination of conidia of both species produced on riboflavin-supplemented medium and exposed to filtered solar radiation was significantly higher than those produced on medium not supplemented with riboflavin. Riboflavin increased the transcription of photolyases, laccases and polyketide synthase genes. However, each fungal species induced different genes patterns involved in DNA repair and photoprotection. The addition of riboflavin to the substrate used for mass production of Metarhizium spp. and the resulting enhancement of conidial tolerance to solar radiation may improve the effectiveness of these fungi in biological control programs.Fil: Huarte Bonnet, Carla. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Pereira Junior, Ronaldo A.. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Santos Da Paixao, Flavia Regina. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Braga, Gilberto U.L.. Universidade de Sao Paulo; BrasilFil: Roberts, Donald W.. State University of Utah; Estados UnidosFil: Luz, Christian. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; BrasilFil: Pedrini, Nicolás. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Fernandes, Éverton. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; Brasi

Relative humidity impacts development and activity against Aedes aegypti adults by granular formulations of Metarhizium humberi microsclerotia

The impact of ambient relative humidity (RH) on conidial production of Metarhizium humberi IP 46 microsclerotia (MS) formulated in pellets or granules was investigated, and a promising granular formulation was tested against Aedes aegypti adults to confirm its efficacy. Microcrystalline cellulose (MC) and diatomaceous earth (DE) or a combination of vermiculite (VE), DE and silicon dioxide (SD) were tested as carriers in granular formulations containing MS. A range of 93?96.5%RH was critical for fungal development, and at least 96.5?98.5% RH was required for high conidial production on pellets or granules. Conidial production was clearly higher on pellets and granules prepared with VE than MC as the main carrier. VE granules containing MS were highly active against A. aegypti adults. Most mosquitoes were killed within 6 days after treatment regardless of the exposure time of adults to the formulation (1 min?24 h) or ambient humidity (75 or >98%). Production of conidia on dead adults varied between 7.3 × 106 and 2.2 × 107 conidia/individual, when exposed to MS granules for 12 h and 1 min, respectively. Granular formulations containing VE as the main carrier and MS as the active ingredient of M. humberi have strong potential for use against A. aegypti.Fil: Rodrigues, Juscelino. Universidade Federal de Goias. Instituto de Patologia Tropical e Saude Publica; BrasilFil: Lopes Catão, Alaine Maria. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; BrasilFil: Soares dos Santos, Amanda. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; BrasilFil: Santos Da Paixao, Flavia Regina. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Rodrigues Santos, Thainá. Universidade Federal de Goiás; BrasilFil: Martinez, Juan Mercado. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; BrasilFil: Marreto, Ricardo Neves. Universidade Federal de Goiás; BrasilFil: Moura Mascarin, Gabriel. Empresa Brasileira de Pesquisa Agropecúaria - Embrapa; BrasilFil: Kort Kamp Fernandes, Éverton. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; BrasilFil: Humber, Richard Alan. United States Department of Agriculture. Agricultural Research Service; ArgentinaFil: Luz, Christian. Universidade Federal de Goias. Instituto de Patologia Tropical E Saude Publica.; Brasi

Inorganic pellets containing microsclerotia of Metarhizium anisopliae: a new technological platform for the biological control of the cattle tick Rhipicephalus microplus

This study was sought to devise pellets containing inorganic materials and microsclerotia of Metarhizium anisopliae strain IP 119 for biological control of Rhipicephalus microplus, the most economically important tick in Brazilian cattle industry. In addition, we evaluated the storage stability of the pellets, their tolerance to ultraviolet radiation (UV-B), and efficacy against ticks under laboratory conditions. Fungal microsclerotia were produced by liquid culture fermentation and mixed with pre-selected inorganic matrices: vermiculite powder, diatomaceous earth, and colloidal silicon dioxide (78:20:2, w/w/w). The microsclerotial pellets were then prepared by a two-stage process involving extrusion and spheronization. Pellet size averaged 525.53 ± 7.74 μm, with a sphericity index of 0.72 ± 0.01, while biomass constituents did not affect the wet mass properties. Conidial production from microsclerotial pellets upon rehydration ranged from 1.85 × 109 to 1.97 × 109 conidia g−1 with conidial viability ≥ 93%. Conidial production from pellets stored at 4 °C was invariable for up to 21 days. Unformulated microsclerotia and microsclerotial pellets were extremely tolerant to UV-B compared with aerial conidia. Engorged tick females exposed to conidia from sporulated pellets applied to soil samples and upon optimal rehydration exhibited shorter oviposition time length, shorter life span, and reduced number of hatched larvae. In summary, microsclerotial pellets of M. anisopliae IP 119 effectively suppressed R. microplus and showed outstanding UV-B tolerance in laboratory tests. Prospectively, this formulation prototype is promising for targeting the non-parasitic stage of this tick on outdoor pasture fields and may offer a novel mycoacaricide for its sustainable management.Fil: Santos, Thainá Rodrigues. Universidade Federal de Goiás; BrasilFil: Santos Da Paixao, Flavia Regina. Universidade Federal de Goiás; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Catão, Alaine Maria Lopes. Universidade Federal de Goiás; BrasilFil: Muniz, Elen Regozino. Universidade Federal de Goiás; BrasilFil: Ribeiro-Silva, Cárita Souza. Universidade Federal de Goiás; BrasilFil: Taveira, Stephania Fleury. Universidade Federal de Goiás; BrasilFil: Luz, Christian. Universidade Federal de Goiás; BrasilFil: Mascarin, Gabriel Moura. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; BrasilFil: Fernandes, Éverton Kort Kamp. Universidade Federal de Goiás; BrasilFil: Marreto, Ricardo Neves. Universidade Federal de Goiás; Brasi