Compatibilidad de un bioinsecticida a base de Beauveria bassiana con agroquímicos: efecto en la germinación y la actividad insecticida sobre Diatraea saccharalis

Los barrenadores del tallo del género de Diatraea spp. (Lepidoptera:Crambidae) han sido reconocidos por ser la plaga de mayor impacto en el cultivo de caña de azúcar, en el que causan importantes pérdidas económicas. Para su control, una cepa colombiana de Beauveria bassiana (Bals.) Vuill (aislamiento Bv062) que demostró alta virulencia frente a larvas de D. saccharalis (Fabricius), fue utilizada para el desarrollo de un bioinsecticida formulado como un concentrado emulsionable (CE). Con el fin de integrar este bioinsecticida en un esquema de manejo integrado de plagas, es necesario evaluar su compatibilidad con los agroquímicos utilizados comúnmente en el cultivo de caña de azúcar. Para tal fin se evaluó el efecto in vitro de 11 agroquímicos (fungicidas, insecticidas y herbicidas) sobre la germinación y actividad insecticida de conidios formulados. Inicialmente el CE y los agroquímicos fueron ajustados al doble de la concentración recomendada y mezclados en proporción 1:1, se incubaron durante dos horas, y se determinó el porcentaje de germinación. Los conidios expuestos a los agroquímicos fueron también purificados y utilizados para un bioen-sayo frente a larvas de segundo instar de D. saccharalis. El CE resultó ser compatible con los herbicidas e insecticidas evaluados, manteniendo una germinación y eficacia superior al 80%. Por el contrario, el CE fue incompatible con todos los fungicidas evaua-dos. Estos resultados permiten sugerir recomendaciones de aplicación del bioplaguicida a base de B. bassiana Bv062, junto con insecticidas y herbicidas empleados en el cultivo de caña, pero se recomienda no aplicarlo de manera simultánea con fungicidas como el benomil y del grupo de triazoles. Se recomienda validar estos resultados en condiciones de campo.

Sugarcane stem borers of the genus Diatraea spp. (Lepidoptera:Crambidae) have been recognized as the pest of the greatest impact on sugarcane crops causing significant economic losses. For its control, a colombian strain of Beauveria bassiana (Bals.) Vuill (code Bv062) that showed a high virulence against D. saccharalis (Fabricius) larvae was used for a biopesticide development, formulated as emulsifiable concentrate (EC). In order to include the EC into an integrated pest management (IPM) scheme, it is necessary to evaluate its compatibility with the agrochemicals commonly used in sugarcane crops. For this purpose, the in vitro effect of 11 agrochemicals (fungicides, insecticides and herbicides) on the germination and insecticidal activity of Bv062 formulated conidia was evaluated. Initially, the EC and agrochemicals were adjusted to double the recommended concentration and mixed in a 1:1 ratio. The mixes were incubated for two hours, and the germination percentage was calculated. The conidia exposed to the agrochemicals were also purified and used for a bioassay against second instar larvae of D. saccharalis. The EC was compatible with the herbicides and insecticides, maintaining germination and efficacy higher than 80%. In contrast, EC was incompatible with all the fungicides evaluated. These results allow us to suggest recommendations for the application of the biopesticide based on B. bassiana Bv062 with insecticides and herbicides used in sugarcane crops, but it is not recommended to apply it simultaneously with fungicides such as benomyl and the triazole group. It is recommended to validate these results under field conditions.

Identification of locally isolated entomopathogenic Fusarium species from the soil of Changa Manga Forest, Pakistan and evaluation of their larvicidal efficacy against Aedes aegypti / Identificação de espécies de Fusarium entomopatogênicas localmente isoladas do solo da Floresta de Changa Manga, Paquistão e avaliação de sua eficácia larvicida contra Aedes aegypti

Dengue fever vectored by the mosquito Aedes aegypti is one of the most rapidly spreading insect-borne diseases. Current reliance of dengue vector control is mostly on chemical insecticides. Growing insecticide resistance in the primary mosquito vector, Aedes aegypti, limits the effectiveness of vector control through chemical insecticides. These chemical insecticides also have negative environmental impacts on animals, plants and human health. Myco-biocontrol agents are naturally occurring organisms and are found to be less damaging to the environment as compared to chemical insecticides. In the present study, entomopathogenic potential of local strains of fungi isolated from soil was assessed for the control of dengue vector. Local fungal isolates presents better alternative to introducing a foreign biocontrol strain, as they may be better adapted to environmental conditions of the area to survive and may have more entomopathogenic efficacy against target organism. Larvicidal efficacy of Fusarium equiseti and Fusarium proliferatum was evaluated against Aedes aegypti. Local strains of F. equiseti (MK371718) and F. proliferatum (MK371715) were isolated from the soil of Changa Manga Forest, Pakistan by using insect bait method. Larvicidal activity of two Fusarium spp. was tested against forth instar larvae of A. aegypti in the laboratory, using concentrations 105, 106, 107 and 108 conidia /ml. LC50 values for F. equiseti after 24h, 48h, 72h and 96h of exposure were recorded as 3.8x108, 2.9x107, 2.0x107, and 7.1x106 conidia /ml respectively while LC50 values for F. proliferatum were recorded as 1.21x108, 9.6x107, 4.2x107, 2.6x107 conidia /ml respectively after 24h, 48h, 72h and 96h of exposure. The results indicate that among two fungal strains F. equiseti was found to be more effective in terms of its larvicidal activity than F. proliferatum against larvae of A. aegypti.

A dengue transmitida pelo mosquito Aedes aegypti é uma das doenças transmitidas por insetos de propagação mais rápida. A dependência atual do controle do vetor da dengue é principalmente de inseticidas químicos. O aumento da resistência a inseticidas no principal vetor do mosquito, Aedes aegypti, limita a eficácia do controle do vetor por meio de inseticidas químicos. Esses inseticidas químicos também têm impactos ambientais negativos sobre os animais, plantas e saúde humana. Os agentes de micobiocontrole são organismos que ocorrem naturalmente e são menos prejudiciais ao meio ambiente em comparação com os inseticidas químicos. No presente estudo, avaliou se o potencial entomopatogênico de cepas locais de fungos isolados do solo para o controle do vetor da dengue. Isolados de fungos locais apresentam melhor alternativa para a introdução de uma cepa de biocontrole estrangeira, pois podem ser mais bem adaptados às condições ambientais da área para sobreviver e podem ter maior eficácia entomopatogênica contra o organismo-alvo. A eficácia larvicida de Fusarium equiseti e Fusarium proliferatum foi avaliada contra Aedes aegypti. Cepas locais de F. equiseti (MK371718) e F. proliferatum (MK371715) foram isoladas do solo de Changa Manga Forest, Paquistão, usando o método de isca para insetos. Atividade larvicida de dois Fusarium spp. foi testado contra larvas de quarto ínstar de A. aegypti em laboratório, nas concentrações 105, 106, 107 e 108 conídios / ml. Os valores de LC50 para F. equiseti após 24 h, 48 h, 72 h e 96 h de exposição foram registrados como 3,8x 108, 2,9x107, 2,0x107 e 7,1x106 conídios / ml, respectivamente, enquanto os valores de LC50 para F. proliferatum foram registrados como 1,21x108, 9,6 x107, 4,2x107, 2,6x107 conídios / ml, respectivamente, após 24 h, 48 h, 72 h e 96 h de exposição. Os resultados indicam que entre duas cepas de fungos F. equiseti se mostrou mais eficaz em termos de atividade [...].

Identification of locally isolated entomopathogenic Fusarium species from the soil of Changa Manga Forest, Pakistan and evaluation of their larvicidal efficacy against Aedes aegypti

Abstract Dengue fever vectored by the mosquito Aedes aegypti is one of the most rapidly spreading insect-borne diseases. Current reliance of dengue vector control is mostly on chemical insecticides. Growing insecticide resistance in the primary mosquito vector, Aedes aegypti, limits the effectiveness of vector control through chemical insecticides. These chemical insecticides also have negative environmental impacts on animals, plants and human health. Myco-biocontrol agents are naturally occurring organisms and are found to be less damaging to the environment as compared to chemical insecticides. In the present study, entomopathogenic potential of local strains of fungi isolated from soil was assessed for the control of dengue vector. Local fungal isolates presents better alternative to introducing a foreign biocontrol strain, as they may be better adapted to environmental conditions of the area to survive and may have more entomopathogenic efficacy against target organism. Larvicidal efficacy of Fusarium equiseti and Fusarium proliferatum was evaluated against Aedes aegypti. Local strains of F. equiseti (MK371718) and F. proliferatum (MK371715) were isolated from the soil of Changa Manga Forest, Pakistan by using insect bait method. Larvicidal activity of two Fusarium spp. was tested against forth instar larvae of A. aegypti in the laboratory, using concentrations 105, 106, 107 and 108 conidia /ml. LC50 values for F. equiseti after 24h, 48h, 72h and 96h of exposure were recorded as 3.8x 108, 2.9x107, 2.0x107, and 7.1x106 conidia /ml respectively while LC50 values for F. proliferatum were recorded as 1.21x108, 9.6x107, 4.2x107, 2.6x107 conidia /ml respectively after 24h, 48h, 72h and 96h of exposure. The results indicate that among two fungal strains F. equiseti was found to be more effective in terms of its larvicidal activity than F. proliferatum against larvae of A. aegypti.

Resumo A dengue transmitida pelo mosquito Aedes aegypti é uma das doenças transmitidas por insetos de propagação mais rápida. A dependência atual do controle do vetor da dengue é principalmente de inseticidas químicos. O aumento da resistência a inseticidas no principal vetor do mosquito, Aedes aegypti, limita a eficácia do controle do vetor por meio de inseticidas químicos. Esses inseticidas químicos também têm impactos ambientais negativos sobre os animais, plantas e saúde humana. Os agentes de micobiocontrole são organismos que ocorrem naturalmente e são menos prejudiciais ao meio ambiente em comparação com os inseticidas químicos. No presente estudo, avaliou-se o potencial entomopatogênico de cepas locais de fungos isolados do solo para o controle do vetor da dengue. Isolados de fungos locais apresentam melhor alternativa para a introdução de uma cepa de biocontrole estrangeira, pois podem ser mais bem adaptados às condições ambientais da área para sobreviver e podem ter maior eficácia entomopatogênica contra o organismo-alvo. A eficácia larvicida de Fusarium equiseti e Fusarium proliferatum foi avaliada contra Aedes aegypti. Cepas locais de F. equiseti (MK371718) e F. proliferatum (MK371715) foram isoladas do solo de Changa Manga Forest, Paquistão, usando o método de isca para insetos. Atividade larvicida de dois Fusarium spp. foi testado contra larvas de quarto ínstar de A. aegypti em laboratório, nas concentrações 105, 106, 107 e 108 conídios / ml. Os valores de LC50 para F. equiseti após 24 h, 48 h, 72 h e 96 h de exposição foram registrados como 3,8x 108, 2,9x107, 2,0x107 e 7,1x106 conídios / ml, respectivamente, enquanto os valores de LC50 para F. proliferatum foram registrados como 1,21x108, 9,6 x107, 4,2x107, 2,6x107 conídios / ml, respectivamente, após 24 h, 48 h, 72 h e 96 h de exposição. Os resultados indicam que entre duas cepas de fungos F. equiseti se mostrou mais eficaz em termos de atividade larvicida do que F. proliferatum contra larvas de A. aegypti.

Identification of locally isolated entomopathogenic Fusarium species from the soil of Changa Manga Forest, Pakistan and evaluation of their larvicidal efficacy against Aedes aegypti / Identificação de espécies de Fusarium entomopatogênicas localmente isoladas do solo da Floresta de Changa Manga, Paquistão e avaliação de sua eficácia larvicida contra Aedes aegypti

Abstract Dengue fever vectored by the mosquito Aedes aegypti is one of the most rapidly spreading insect-borne diseases. Current reliance of dengue vector control is mostly on chemical insecticides. Growing insecticide resistance in the primary mosquito vector, Aedes aegypti, limits the effectiveness of vector control through chemical insecticides. These chemical insecticides also have negative environmental impacts on animals, plants and human health. Myco-biocontrol agents are naturally occurring organisms and are found to be less damaging to the environment as compared to chemical insecticides. In the present study, entomopathogenic potential of local strains of fungi isolated from soil was assessed for the control of dengue vector. Local fungal isolates presents better alternative to introducing a foreign biocontrol strain, as they may be better adapted to environmental conditions of the area to survive and may have more entomopathogenic efficacy against target organism. Larvicidal efficacy of Fusarium equiseti and Fusarium proliferatum was evaluated against Aedes aegypti. Local strains of F. equiseti (MK371718) and F. proliferatum (MK371715) were isolated from the soil of Changa Manga Forest, Pakistan by using insect bait method. Larvicidal activity of two Fusarium spp. was tested against forth instar larvae of A. aegypti in the laboratory, using concentrations 105, 106, 107 and 108 conidia /ml. LC50 values for F. equiseti after 24h, 48h, 72h and 96h of exposure were recorded as 3.8x 108, 2.9x107, 2.0x107, and 7.1x106 conidia /ml respectively while LC50 values for F. proliferatum were recorded as 1.21x108, 9.6x107, 4.2x107, 2.6x107 conidia /ml respectively after 24h, 48h, 72h and 96h of exposure. The results indicate that among two fungal strains F. equiseti was found to be more effective in terms of its larvicidal activity than F. proliferatum against larvae of A. aegypti.

Resumo A dengue transmitida pelo mosquito Aedes aegypti é uma das doenças transmitidas por insetos de propagação mais rápida. A dependência atual do controle do vetor da dengue é principalmente de inseticidas químicos. O aumento da resistência a inseticidas no principal vetor do mosquito, Aedes aegypti, limita a eficácia do controle do vetor por meio de inseticidas químicos. Esses inseticidas químicos também têm impactos ambientais negativos sobre os animais, plantas e saúde humana. Os agentes de micobiocontrole são organismos que ocorrem naturalmente e são menos prejudiciais ao meio ambiente em comparação com os inseticidas químicos. No presente estudo, avaliou-se o potencial entomopatogênico de cepas locais de fungos isolados do solo para o controle do vetor da dengue. Isolados de fungos locais apresentam melhor alternativa para a introdução de uma cepa de biocontrole estrangeira, pois podem ser mais bem adaptados às condições ambientais da área para sobreviver e podem ter maior eficácia entomopatogênica contra o organismo-alvo. A eficácia larvicida de Fusarium equiseti e Fusarium proliferatum foi avaliada contra Aedes aegypti. Cepas locais de F. equiseti (MK371718) e F. proliferatum (MK371715) foram isoladas do solo de Changa Manga Forest, Paquistão, usando o método de isca para insetos. Atividade larvicida de dois Fusarium spp. foi testado contra larvas de quarto ínstar de A. aegypti em laboratório, nas concentrações 105, 106, 107 e 108 conídios / ml. Os valores de LC50 para F. equiseti após 24 h, 48 h, 72 h e 96 h de exposição foram registrados como 3,8x 108, 2,9x107, 2,0x107 e 7,1x106 conídios / ml, respectivamente, enquanto os valores de LC50 para F. proliferatum foram registrados como 1,21x108, 9,6 x107, 4,2x107, 2,6x107 conídios / ml, respectivamente, após 24 h, 48 h, 72 h e 96 h de exposição. Os resultados indicam que entre duas cepas de fungos F. equiseti se mostrou mais eficaz em termos de atividade larvicida do que F. proliferatum contra larvas de A. aegypti.

Biocontrol potential of Beauveria bassiana and Metarhizium anisopliae isolates from Turkey against Hyphantria cunea (Drury) (Lepidoptera: arctiidae) Larvae under laboratory and field conditions

Hyphantria cunea is one of the most important pest insects causing significant damage in many plant species. The present study aimed to evaluate the insecticidal activity of Turkey isolates of Beauveria bassiana and Metarhizium anisopliae against H. cunea larvae under laboratory and field conditions. B. bassiana isolates YK16, YK23 and YK26, and M. anisopliae isolates YK41 and YK45 were sprayed onto the larvae of H. cunea at the respective doses of 1x105 and 1x106 conidia mL-1 and monitored for seven days. Fungal isolates, bio-insecticide Bacillus thuringiensis and chemical insecticide diflubenzuron were used in field studies. Trials were carried out in five replications. All B. bassiana isolates caused 100% mortality on first instar larvae in laboratory trials. Mortality ratios ranged from 100 to 96% on second instar larvae. M. anisopliae isolates YK45 and YK41 caused 88 and 84%, and 81.33 and 77.11% mortalities for the first and second instar larvae, respectively. The mortality rates fluctuated between 91.78-72.89% for B. bassiana on third instar larvae in laboratory conditions. However, M. anisopliae isolates YK45 and YK41 caused 77.11 and 60.22% mortality on third instar larvae, respectively. In the field trials, B. bassiana YK23 displayed promising insecticidal activity with 80.60% mortality on second instar larvae of H. cunea. Other isolates as well caused mortalities ranging from 60.77 to 49.55%. The results revealed that some isolates of B. bassiana and M. anisopliae have potential to control H. cunea larvae. However, additional detailed studies need to be carried out to increase their effectiveness in field conditions.

Identification of entomopathogenic fungi Metarhizium anisopliae and Purpureocillium lilacinum from oil palm plantation soils in Universiti Putra Malaysia

Aims@#Insect pests are one of the major constraints of oil palm production in Malaysia. However, synthetic chemical insecticides are the most common method for its control, despite their negative effects on non-target organisms and the development of resistance. Therefore, the present study is aimed to identify novel indigenous isolates of entomopathogenic fungi (EPF) in oil palm soil as part of integrated pest management (IPM) of oil palm insect pests.@*Methodology and results@#The potential of EPF were isolated from the soil collected from the oil palm plantation in UPM using a mealworm beetle larva (Tenebrio molitor) as an insect bait. Seven Metarhizium anisopliae and two Purpureocillium lilacinum isolates were identified by morphological characterization (macroscopic and microscopic observation) and molecular identification using internal transcribed spacer (ITS) region (ITS region amplification).@*Conclusion, significance and impact of study@#To our knowledge, this is the first time P. lilacinum has been found in the soil of an oil palm plantation or any other host in Malaysia. Furthermore, both of the isolates M. anisopliae and P. lilacinum may be potentially considered as biological control candidates for major insect pests in oil palm.

CONTROL DE Aedes aegypti (DIPTERA: CULICIDAE) MEDIANTE ACTINOBACTERIAS FORMADORAS DE BIOPELÍCULAS / Control of Aedes aegypti (Díptera: Culicidae) by means of biofilm-forming actinobacteria

RESUMEN Las actinobacterias incluyen miembros productores de compuestos multipropósitos restringidamente sobreexplotados al género Streptomyces. No obstante, es necesario reorientar la exploración de bacterias no estreptomiceas para el uso de nuevos bioagentes ecológicos con miras a evitar redescubrimientos y contrarrestar la resistencia a insecticidas químicos en Aedes aegypti. En esta investigación, se caracterizó actinobacterias formadoras de biopelículas para evaluar su dinámica de crecimiento, actividad larvicida y efectos subletales. La identificación, crecimiento y bioactividades de los patógenos se realizaron por cultivos, análisis de imágenes por fotomicrografía y bioensayos. Las biopelículas pertenecen a Pseudonocardiaceae (PsA1TA) y Corynebacteriaceae (CoA2CA) característicamente dependientes a crecer sobre sustratos con revestimiento cuticular específico. PsA1TA coloniza estructuras membranosas de tórax y abdomen de larvas. Las microcolonias desarrollan infectivamente a biopelículas mono y biestratificadas hasta cubrir cuatro veces la amplitud toracoabdominal (envergadura infectiva entre 1010 μm a 1036 μm). En contraste, CoA2CA envuelve radialmente estructuras esclerotizadas cefálica y anal al triplicar la amplitud de los órganos infectados (envergadura infectiva: 1820 a 2030 μm y 1650 a 1860 μm, respectivamente). Las biopelículas ejercen mortalidad diferenciada a todas las etapas larvarias, no obstante, PsA1TA resultó más mortal y virulento frente al segundo estadio (58 %-96 horas, TL50: 3,4 días), mientras que CoA2CA lo fue sobre el cuarto estadio larval (85 %-96 horas, TL50: 2,5 días). CoA2CA indujo emergencia incompleta de farados y despliegue de tarsos curvos en adultos emergentes, además de revestir cadáveres larvarios con robustas biopelículas. Los morfotipos actinobacterianos revelan efecto larvicida y subletal en A. aegypti por formación de biopelículas.

ABSTRACT Actinobacteria include several ubiquitous members involved in the synthesis of multipurpose bioactive compounds strictly derived from the genus Streptomyces. Nevertheless, new bacterial consortia based on non-streptomycetes actinobacteria are needed to be explored in order to avoid rediscoveries and minimize the development of insecticide resistance in Aedes aegypti. In accordance with the use of eco-friendly bioagents, in this study biofilm-forming actinobacteria were characterized on the basis of assessment their growth dynamics, larvicidal mortality and sublethal effects. Actinobacteria identification, biofilm growth and larvicidal bioactivities were performed by employing bacterial cultures, photomicrograph-based image analysis and bioassays. Bacterial morphotypes belong to Pseudonocardiaceae (PsA1TA) and Corynebacteriaceae (CoA2CA), which showed a distinctly substrate-dependent growth. PsA1TA microcolonies were randomly distributed on abdominal and thoracic membranous epicuticle. Afterwards, the thickness of mono-and bi-layered biofilms were increased fourfold the larval thoracoabdominal width (infectious breadth, 1010 μm - 1036 μm). In contrast, cephalic and anal sclerotized structures were radially encased by CoA2CA biofilms and increased threefold the size of both structures (infectious breadth, 1820 - 2030 μm y 1650 - 1860 μm, respectively). Although biofilms caused dissimilar mortality rates on the four larval instars, PsA1TA exerted highest larvicidal activity and virulence on second instar larvae (58 %-96 hours, LT50: 3.4 days) y CoA2CA on fourth instar larvae (85 %-96 hours, LT5G: 2.5 days). CoA2CA also induced incomplete release of pharate individuals as well as buckled protruding tarsi in newly emergent adults, whilst larval cadavers were overwhelmingly encased within massive biofilm aggregates. Biofilm-forming actinobacteria performed biolarvicidal activity and sublethal responses in A. aegypti.

Isolates of Bacillus thuringiensis from Maranhão biomes with potential insecticidal action against Aedes aegypti larvae (Diptera, Culicidae) / Isolados de Bacillus thuringiensis de biomas maranhenses com potencial ação inseticida contra larvas de Aedes aegypti (Diptera, Culicidae)

Abstract Entomopathogenic agents are viable and effective options due to their selective action against insects but benign effects on humans and the environment. The most promising entomopathogens include subspecies of Bacillus thuringiensis (Bt), which are widely used for the biological control of insects, including mosquito vectors of human pathogens. The efficacy of B. thuringiensis toxicity has led to the search for new potentially toxic isolates in different regions of the world. Therefore, soil samples from the Amazon, Cerrado and Caatinga biomes of the state of Maranhão were evaluated for their potential larvicidal action against Aedes aegypti. The isolates with high toxicity to mosquito larvae, as detected by bioassays, were subjected to histological evaluation under a light microscope to identify the genes potentially responsible for the toxicity. Additionally, the toxic effects of these isolates on the intestinal epithelium were assessed. In the new B. thuringiensis isolates toxic to A. aegypti larvae, cry and cyt genes were amplified at different frequencies, with cry4, cyt1, cry32, cry10 and cry11 being the most frequent (33-55%) among those investigated. These genes encode specific proteins toxic to dipterans and may explain the severe morphological changes in the intestine of A. aegypti larvae caused by the toxins of the isolates.

Resumo Os agentes entomopatógenos são alternativas viáveis e eficazes, devido à sua ação seletiva para insetos sendo inofensivos ao homem e ao meio ambiente. Dentre os entomopatógenos mais promissores, destacam-se as subespécies de Bacillus thuringiensis (Bt) amplamente utilizadas no controle biológico de insetos incluindo espécies de mosquitos vetores de agentes patogênicos ao homem. A eficiência da toxicidade de Bt incentiva a prospecção de novos isolados em diversas regiões do mundo. Desta forma, em busca de novos isolados de B. thuringiensis potencialmente tóxicos, amostras de solo provenientes dos biomas Amazônia, Cerrado e Caatinga do estado do Maranhão foram avaliadas em relação ao seu potencial larvicida para Aedes aegypti. Os isolados que provocaram elevada toxicidade para larvas do mosquito, detectada por bioensaios, foram avaliados em relação aos potenciais genes responsáveis pela atividade tóxica, além da avaliação de efeitos tóxicos no epitélio intestinal através de análises histológicas em microscopia de luz. Os novos isolados de Bt tóxicos para larva de A. aegypti amplificaram frequências diferentes de genes cry e cyt sendo os mais frequentes (55-33%) os cry4, cyt1, cry32, cry10 e cry11 dentre os investigados. Esses genes codificam para proteínas tóxicas específicas para ordem Diptera, e podem explicar as severas alterações morfológicas provocadas pelas toxinas dos isolados observadas no intestino das larvas de A. aegypti.

Ovicidal Efficacy of Metarhizium anisopliae (Hypocreales: Clavicipitaceae) towards Rhipicephalus sanguineus (Acari: Ixodidae) Eggs

@#Rhipicephalus sanguineus, commonly known as brown dog tick is a widespread species with considerable public health and economic importance. Tremendous efforts were performed to control the tick populations with the concern of resistance build-up and environmental issues. Alternative towards microbial control thus emerged as one option to reduce tick populations. In this study, the ovicidal efficacy of a native isolate entomopathogenic hyphomycetes fungi, Metarhizium anisopliae strain HSAH5 was evaluated against eggs of R. sanguineus. Spray applications with three different conidial concentrations of 105, 106 and 107 conidia mL-1; 40 ppm of Flumethrin and a negative control. The M. anisopliae strain was found highly virulent to R. sanguineus eggs by reducing the hatching percentages to ≈30% compared with 8.9% in Flumethrin eggs. The result showed a significantly higher mortality in M. anisopliae group than those of the control groups (F = 42.08, df = 32, P < 0.001) at 30 days post-infection. However, there are no significant differences within the M. anisopliae group, in which the mortality between different conidial concentrations is almost the same. The estimated LC50 of M. anisopliae against eggs of R. sanguineus is 1.36 × 103 conidia ml-1. Thus, these results suggest M. anisopliae strain HSAH5 could be a potential biocontrol agent of R. sanguineus in the integrated approach to managing ticks in the residential landscape by targeting on the eggs.

Effects of nutrient additives and incubation period on sporulation and viability of the entomopathogenic fungus, Metarhizium anisopliae (Hypocreales: Clavicipitaceae)

Aims@#Metarhizium anisopliae is an entomopathogenic fungus (EPF) that exists naturally in the environment and potentially be used as a biological control agent against many insect pests. This study aims to evaluate the effect of nutrient additives on the yield and viability of M. anisopliae spore and to determine the optimum incubation period for maximum spore production.@*Methodology and results@# In this study, M. anisopliae was cultivated by solid-state fermentation using rice as a growth medium. Three different nutrient additives were examined which aimed to maximize the production of M. anisopliae spores. Among the three nutrient additives evaluated, yeast (1.84 ± 0.04 g) supported better growth and spore production than molasses (0.58 ± 0.04 g) and palm oil (0.47 ± 0.09 g). The incubation period between 2-6 weeks produced higher spore yield (0.97 ± 0.02 g spores) at week 4 with a better spore viability (86.30 ± 0.45%) at week 2. @*Conclusion, significance and impact of study@#Hence, it is suggested that the optimum incubation period is between 2 and 6 weeks after inoculation, and M. anisopliae could be mass produced in large quantities on rice substrate with the addition of yeast as the nutrient additives.

First report of Lema bilineata Germar (Coleoptera: Chrysomelidae) damaging Physalis peruviana (L.) in Brazil / Primeiro relato de Lema bilineata Germar (Coleoptera: Chrysomelidae) causando dano em Physalis peruviana (L.) no Brasil

ABSTRACT: Increased production of the Cape gooseberry (Physalis peruviana L.) in Brazil has given rise to interest in identifying the phytophagous species that might damage this crop to inform preventive control and integrated pest management strategies. In this study, we report the occurrence and describe the damage that larvae and adults of Lema bilineata Germar (Coleoptera: Chrysomelidae) cause in P. peruviana. The number of L. bilineata individuals, both larvae and adults, significantly affected the total consumption of P. peruviana leaves. We also report, for the first time, three natural enemies, including a fungus, a fly, and an ant, which are associated with this pest in Brazil and may play a role in biological control strategies.

RESUMO: Fisális, Physalis peruviana L., é uma cultura em expansão no Brasil, dessa forma a identificação de espécies fitófagas que causam danos nesta cultura é importante para desenvolver controle preventivo e estratégias para o Manejo Integrado de Pragas. Neste estudo, relatamos a ocorrência de Lema bilineata Germar (Coleoptera: Chrysomelidae) no cultivo de P. peruviana e descrevemos os danos causados por suas larvas e adultos. Observou-se que o número de indivíduos de L. bilineata, tanto larvas como adultos, influenciaram significativamente o consumo total de folhas de P. peruviana. Além disso, relatamos pela primeira vez a presença de três inimigos naturais, incluindo um fungo, uma mosca e uma formiga, associados a esta praga no Brasil que poderiam ser utilizados como agentes de controle biológico.

Dual effects of entomopathogenic fungi on control of the pest Lobesia botrana and the pathogenic fungus Eutypella microtheca on grapevine

BACKGROUND: Entomopathogenic fungi (EPF) are the natural enemies of insect pests. Nevertheless, research on the use of EPF for simultaneous prevention of pest and disease agents on the same crop is limited. In this study, we explored the potential dual effects of three strains of the EPF Metarhizium anisopliae on the control of detrimental agents of Vitis vinifera L., including different developmental stages (larvae, pupae, and adult) of the insect pest Lobesia botrana and the phytopathogenic fungus Eutypella microtheca. METHODS: Laboratory pathogenicity trials were performed to examine the effects of the three M. anisopliae strains on the mortality rate of L. botrana. In addition, field trials were conducted to assess the biocontrol potential of one selected M. anisopliae strain on the larval stage of L. botrana. Moreover, inhibitory effects of the three EPF strains on E. microtheca growth were examined in vitro. RESULTS: All the M. anisopliae strains were highly effective, killing all stages of L. botrana as well as inhibiting the growth of E. microtheca. The in vitro mortality of larvae treated with the strains was over 75%, whereas that of treated pupae and adults was over 85%. The three EPF strains showed similar efficacy against larvae and adult stages; never-theless, pupal mortality was observed to be strain dependent. Mortality of L. botrana larvae ranged from 64 to 91% at field conditions. Inhibition of E. microtheca growth reached 50% in comparison to the control. CONCLUSIONS: Our study showed that M. anisopliae strains were highly effective in ensuring control of two different detrimental agents of V. vinifera L., providing new evidence to support the dual effects of entomopathogenic fungi.

Growth Pattern of Beauveria bassiana in Different Eco-friendly Media

Beauveria bassiana belongs to the class Deuteromycetes which is one of the important disease-causing biocontrol agents in insects. B. bassiana, formerly known as Botrytis bassiana (Balsamo), is a widely distributed soil inhabiting fungus. B. bassiana is also known to be best biocontrol agent against larval stage of the silkworm. B. bassiana is categorized as a white muscardine fungus due to the white color of sporulating colonies. It is a type of biopesticide which is based on entomopathogenic fungi which are often considerable scope as plant protection agents against several pathogens and insects including whiteflies, aphids, thrips, grasshoppers, and certain types of beetles. The present study deals with the use of different media such as coconut media, jaggery media, nutrient media, potato dextrose media, Sabouraud dextrose media, and molasses media

Alternative substrates for conidiogenesis of the entomopathogenic fungus Beauveria bassiana (Bals) Vuillemin (Deuteromycotina: Hyphomycetes) / Substratos alternativos para conidiogênese do fungo entomopatogênico Beauveria bassiana (Bals) Vuillemin (Deuteromycotina: Hyphomycetes)

Abstract Beauveria bassiana is a promising fungus for the biological control of insect pests. The growing costs of conidia production have raised the need to ascertain the efficiency of some low cost substrates. The aim of this study was to analyze the potential use of different raw substrates without nutritional supplement for B. bassiana conidiogenesis. Growth and sporulation were evaluated using 30 g of substrate and 0.3 μL of a conidia suspension (1 x 106 conidia/mL). After 10 days of incubation (70 ± 10% humidity and temperature (T) = 29 ± 1 °C), rice (2.00 x 106 conidia/g substrate), algaroba (2.36 x 106 conidia/g), malt A (1.22 x 106 conidia/g) and malt B (1.75 x 106 conidia/g) showed the highest levels of conidia production. The resulting conidia showed insecticidal activity higher than 80% on coconut termites. These new raw substrates may represent viable alternatives for the production of entomopathogenic fungi for use in the biological control of various insect pests.

Resumo Beauveria bassiana é um fungo promissor no controle biológico de insetos-praga. As crescentes despesas na produção de conídios levantam a necessidade de averiguar a eficiência de alguns substratos de baixo custo. O objetivo deste trabalho foi analisar o potencial de utilização de diferentes substratos brutos para a conidiogênese de B. bassiana. O crescimento e esporulação foram realizados utilizando 30 g do substrato e 0,3 µL da suspensão de conídios (1 x 106 conídios/mL). Após 10 dias de incubação (umidade 70 ± 10% e temperatura T = 29 ± 1° C), o arroz (2,00 x 106 conídios/g de substrato), algaroba (2,36 x 106 conídios/g), malte A (1,22 x 106 conídios/g) e B (1,75 x 106 conídios/g), apresentaram maior produção de conídios. Os conídios produzidos mostraram atividade inseticida sobre o cupim do coqueiro acima de 80% de mortalidade. Estes novos substratos brutos podem representar uma alternativa viável para produção de fungos entomopatogênicos para uso no controle biológico de vários insetos praga.

In vitro efficacy of two commercial products of Metarhizium anisopliae s.l. for controlling the cattle tick Rhipicephalus microplus / Eficácia in vitro de dois produtos comerciais de Metarhizium anisopliae s.l. no controle do carrapato bovino Rhipicephalus microplus

Abstract The effects of two different products - Metarril® SP Organic (dry conidia) and Metarril® SC Organic (emulsifiable concentrated conidia in vegetable oil) - on eggs, larvae and Rhipicephalus microplus engorged females were here explored. Three concentrations (108, 107, and 106 conidia mL-1) for both products were prepared in water + 0.1% Tween® 80 (v/v); afterward, bioassays were carried out for all R. microplus stages by immersion in suspensions (Metarril® SP) or formulations (Metarril® SC). Metarril® SP suspensions showed low efficacy and did not affect biological parameters of treated engorged females; for eggs and larvae, only slight decreases in hatchability and larvae population were observed. Despite a delay in germination, Metarril® SC presented better results; for females, reductions in Egg Mass Weight (EMW) and Egg Production Index (EPI) were reported. On eggs, 108 conidia mL-1 increased Incubation Period (IP), shortened Hatching Period (HP) and decreased hatchability by up to 61%; for larvae, 107 and 108 conidia mL-1 reached 99.6 and 100% larval mortality respectively, 10 days after fungal exposure. Thus, further studies involving the use of oil-based formulations for ticks such as Metarril® SC need to be performed, especially to control the most susceptible stages (eggs and larvae).

Resumo No presente trabalho, os efeitos de dois diferentes produtos foram avaliados - Metarril® SP Organic (conídios secos) e Metarril® SC Organic (conídios concentrados em óleo vegetal) - para ovos, larvas e fêmeas ingurgitadas de Rhipicephalus microplus. Três concentrações (108, 107 e 106 conídios mL-1) para cada produto foram preparadas em água + Tween® 80 0,1% (v/v); os bioensaios foram realizados para todos os estágios de R. microplus por imersão nas suspensões (Metarril® SP) ou formulações (Metarril® SC). Metarril® SP não afetou os parâmetros biológicos das fêmeas, demonstrando assim baixa eficácia; para ovos e larvas, foram observadas discretas diminuições na eclodibilidade e na população de larvas. Apesar de um atraso na germinação, Metarril® SC apresentou melhores resultados; para as fêmeas, foram detectadas reduções no Peso da Massa de Ovos (PMO) e no Índice de Produção de Ovos (IPO). Para os ovos, a concentração de 108 conídios mL-1 aumentou o Período de Incubação (PI), reduziu o Período de Eclosão (PE) e também o da eclodibilidade em até 61%; para larvas, 107 e 108 conídios mL-1 atingiram 99,6 e 100% de mortalidade larval, respectivamente, 10 dias após a exposição fúngica. Com isso, estudos adicionais que envolvem o uso de formulações à base de óleo para carrapatos, como Metarril® SC, precisam ser realizados, especialmente para controlar os estágios mais suscetíveis (ovos e larvas).

Molecular characterization of Bacillus thuringiensis strains to control Spodoptera eridania (Cramer) (Lepidoptera: Noctuidae) population

Abstract The main objective of this study was to characterize the toxicity and genetic divergence of 18 Bacillus thuringiensis strains in the biological control of Spodoptera eridania. Bacterial suspensions were added to the S. eridania diet. Half of the selected B. thuringiensis strains caused high mortality seven days after infection. The genetic divergence of B. thuringiensis strains was assessed based on Enterobacterial Repetitive Intergenic Consensus (ERIC) and Repetitive Extragenic Palindromic (REP) sequences, and five phylogenetic groups were formed. Despite their genetic diversity B. thuringiensis strains did not show any correlation between the collection sites and toxicity to larvae. Some B. thuringiensis strains are highly toxic to S. eridania, thus highlighting the potential of their endotoxins as biopesticides.

Conidia production of Isaria fumosorosea (Hypocreales: Cordycipitaceae) in organic substrates through two propagation techniques / Produção de conídios de Isaria fumosorosea (Hypocreales: Cordycipitaceae) en substratos orgânicos atraves de duas técnicas de propagação

In the last decades, the use of entomopathogenic fungi for the control of pest insects has increased globally, however a key step to achieve the successful application of fungal propagules as biocontrol agents depends of various factors, inoculum production being one of the essential stages for these microorganisms to be used in pest management programs. The objective of this study was to evaluate theproduction of conidia by solid fermentation and biphasic culture in different sporulation matrices for four native isolates and a strain of Isaria fumosorosea that have been shown potential for the control of various insects at the laboratory level. The experimental design was completely randomized, two propagation methods were used with six treatments and ten repetitions per treatment; the data were analyzed using an analysis of variance and the means were compared using the Tukey test (p≤ 0.05). In the production by solid fermentation the minimum value of 5.30 × 105 conidia g-1 was registered with the pericarp of peanut and the maximum of 2.35 × 107 conidia g-1 in the corn grain; in the biphasic culture the minimum of 7.60 × 105 conidia g-1 was observed in birdseed and the maximum of 2.07 × 107 conidia g-1 in rice. The differences were significant (p ≤0.05) in the production by method and by substrate, in solid fermentation 6.84 × 106 conidia g-1 and in biphasic culture 8.85 × 106 conidia g-1. In the production by substrate, the rice showed 1.75 × 107 conidia g-1 and the lesser canary seed concentration (7.80 × 105 conidia g-1). The average production per isolate and / or strain was of the order of 106 and significant difference was registered (p ≤ 0.05) among the fungi, the isolate HIB-9 showed concentration in average higher in the production (7.90 × 106 conidia g-1) and the isolated HIB-19 was the least effective, with only 1.08 × 106 conidia g-1. The results obtained show marked differences between the isolates with respect to their capacity to use the different substrates used as propagation media to obtain conidia of I. fumosorosea.

Nas últimas décadas, o uso de fungos entomopatogênicos para o controle de insetos-pragatem aumentado globalmente, porém um passo fundamental para o sucesso da aplicação de propágulos fúngicos como agentes de biocontrole depende de varios fatores, sendo a produção de inóculo um dos estágios essenciais para esses insetos microorganismos a serem usados em programas de controle de pragas. O objetivo deste trabalho foi avaliar a produção de conídios por meio de fermentação sólida e cultura bifásica emdiferentes matrizes de esporulação para quatro isolados nativos e um alinhagem de Isaria fumosorosea que tem demonstrado potencial para o controle de varios insetos em nível laboratorial. O delineamento experimental foi inteiramente casualizado, foram utilizados dois métodos de propagação com seis tratamentos e dezrepetições por tratamento; os dados foraman alisados por meio de análise de variância e as médias comparadas pelo teste de Tukey (p ≤ 0.05). Naprodução por fermentação sólida foi registrado o valor mínimo de 5.30 × 105 conídios g-1 com o pericarpo de amendoim e o máximo de 2.35 × 107 conídios g-1 no grão de milho; na cultura bifásica, o mínimo de 7.60 × 105 conídios g-1 foi observado emsementes de aves e o máximo de 2.07 × 107conídios g-1em arroz. As diferençasforam significativas (p ≤ 0.05) na produção pelo método e pelo substrato, na fermentação sólida 6.84 × 106 conídios g-1 e na cultura bifásica 8.85 × 106 conídios g-1. Naprodução por substrato, o arroz apresentou 1.75 × 107conídios g-1 e a menor concentração de sementes de canário (7.80 × 105conídios g-1). A produção média por isolado e / ou cepa foi da ordem de 106 e registrou-se diferença significativa (p ≤ 0.05) entre os fungos, o isolado HIB-9 apresentou concentração em média maior na produção (7.90 × 106conídios g-1) e o isolado HIB-19 foi o menos efetivo, com apenas 1.08 × 106 conídios g-1. Os resultados obtidos mostram diferenças marcantes entre os isolados quanto à capacidade de utilização dos diferentes substratos utilizados como meio de propagação para obtenção de conídios de I. fumosorosea.

Bioformulado de Beauveria bassiana (ATCC MYA-4886) y Trichoderma lignorum (ATCC-8751) como biocontrolador de Atta cephalotes / Bioinsecticide study for controlling the carrier ant (Atta cephalotes), using filamentous fungi spores Beauveria bassiana (ATCC MYA-4886) and Trichoderma lignorum (ATCC-8751) / Bioformulação de Beauveria bassiana (ATCC MYA-4886) e Trichoderma lignorum (ATCC-8751) como biocontrolador de Atta cephalotes

Resumen I. La hormiga arriera está asociada a pérdidas en el sector agrícola, debido a su actividad defoliadora. El control de la especie se ha venido realizado artesanal, química y biológicamente, esta última con beneficios ambientales y de bajo riesgo para la salud humana. El objetivo de esta investigación fue desarrollar una formulación biológica para el control de la hormiga arriera (Atta cephalotes) utilizando una mezcla de esporas de dos hongos filamentosos (Beauveria bassiana y Trichoderma lignorum). M. Se desarrollaron 5 formulaciones empleando las relaciones: 1:1,6:4,4:6,3:7,2:8, de cepas de B. bassiana (ATCC MYA-4886) y T. lignorum (ATCC 8751), realizándoles prueba de viabilidad, patogenicidad y pureza. La colonización de las esporas en tejidos, se evaluó mediante la exposición de ratas Wistar a la formulación y sus componentes, realizando diagnóstico veterinario (disección) y cultivo microbiológico. R. Las formulaciones presentaron viabilidad a 24 h del 95+2 %, el 100% de las formulaciones no se contaminaron después de 10 días, las formulaciones 6.4, 1:1, 2:8 infectaron la totalidad de los individuos en 6 días, las formulaciones 4:6 y 3:7 a los 8 días, no se observó colonización de las cepas en la formulación, ni en tejidos de los biomodelos. C. Las formulaciones 6.4, 1:1, 2:8 de Beauveria bassiana y Trichoderma lignorum, poseen mayor actividad infecciosa sobre la hormiga arriera (Atta cephalotes).

Abstract I. The Leaf cutting Ant is associated with losses in the agricultural sector for the most part in the cultivation of citrus fruit sector due to its activity defoliator Control of the species has been handmade, chemical and biologically, the latter with environmental benefits and low risk to human health. This research had as objective develop a formulation biological for the control of the Leaf cutting Ant (Atta cephalotess using a mixture of spores of two fungi filamentous (Beauveria bassiana and Trichoderma lignorum), M. He was the isolation of Beauveria bassiana (ATCC MYA-4886) and Trichoderma lignorum (ATCC 8751), through cultivation YPDA and was conducted identifying fungal imprint and biochemical tests. Developing five formulations with ratios of 1:1, 6:4, 4:6, 3:7, 2:8 of Beauveria bassiana and Trichoderma lignorum respectively they underwent the test of viability in nutrient agar pathogenicity by immersion test and proof of purity; the tests were performed in triplicate. R. The formulations presented viability to 24 h 95% 2, I00% of the formulations were pure after 10 days, formulations 6.4, 2:8 infected all of the individuals in 6 days, formulations 4:6 and 3:7-8 days of exposure. C. Formulations 6.4, 2:8 of Beauveria bassiana and Trichoderma lignorium, present infectious activity on the Leaf cutting Ant (Atta cephalotess in laboratory.

Resumo I. O formigueiro está associado a perdas no setor agrícola, devido à sua atividade desfolhadora. O controle das espécies tem sido realizado artesanalmente, quimicamente e biologicamente, este último com benefícios ambientais e baixo risco para a saúde humana. O objetivo desta pesquisa foi desenvolver uma formulação biológica para o controle de formigas (Atta cephalotess usando uma mistura de esporos de dois fungos filamentosos (Beauveria bassiana e Trichoderma lignorum), M. 5 As formulações foram desenvolvidas usando a relação: 1 : 1,6: 4,4: 6,3: 7,2: 8 estirpes de B, bassiana (ATCC MAA-4886) e T. lignorum (ATCC 875I), realizándoles Teste de viabilidade, patogenicidade e pureza. A colonização dos esporos nos tecidos foi avaliada pela exposição de ratos Wistar à formulação e seus componentes, realizando diagnóstico veterinário (dissecção) e cultura microbiológica. R. As formulações mostraram viabilidade em 24 h de 95 + 2%, 100% das formulações não foram contaminadas após I0 dias, as formulações 6,4, 1: 1, 2: 8 infectaram todos os indivíduos em 6 dias, as formulações 4: 6 e 3: 7 aos 8 dias, nenhuma colonização das cepas foi observada na formulação, nem nos tecidos dos biomodelos. C. As formulações 6.4, 1: 1, 2: 8 de Beauveria bassiana e Trichoderma lignorum, apresentam maior atividade infecciosa sobre os antirretera (Atta cephalotess).

AMAZONIAN ISOLATES OF Metarhizium ARE EFFECTIVE FOR KILLING Bactrocera carambolae (DIPTERA: TEPHRITIDAE) / Aislados Amazónicos de Metarhizium son efectivos para matar Bactrocera carambolae (Diptera: Tephritidae)

ABSTRACT Bactrocera carambolae Drew & Hancock is a quarantine pest present in Brazil and is the main phytosanitary barrier for the export of fresh fruits from that country. In this work, we evaluated the effect of Amazonian isolates of Metarhizium on kill larvae, pupae, and adults of B. carambolae in soil applications. The mortality of larvae and pupae in sterile soil treated with Metarhizium anisopliae was 70 %. In addition, 100 % of the adults that emerged from this substrate died up to five days later. This isolate caused the mortality of larvae and pupae in non-sterile soil, but its effect was more evident after adult emergence (70 % mortality up to ten days after emergence). Metarhizium robertsii was effective in killing larvae and pupae of the pest, especially in non-sterile soil, and caused a 60 % reduction in adult survival 50 days after emergence. These results indicated the possible use of these isolates for the control of B. carambolae, suggesting the possibility of using this biocontrol mode as another alternative in controlling this quarantine pest.

RESUMEN Bactrocera carambolae Drew & Hancock es una plaga cuarentenaria presente en Brasil y es la principal barrera fitosanitaria para la exportación de frutas frescas de este país. En este trabajo, evaluamos el efecto de los aislados amazónicos de Metarhizium para matar larvas, pupas y adultos de B. carambolae en aplicaciones de suelo. La mortalidad de larvas y pupas en suelo estéril tratado con Metarhizium anisopliae fue del 70 %. Además, el 100 % de los adultos que emergieron de este sustrato murieron hasta cinco días después. Este aislado causó la mortalidad de larvas y pupas en suelo no estéril, pero su efecto fue más evidente después de la emergencia del adulto (70 % de mortalidad hasta diez días después de la emergencia). Metarhizium robertsii fue eficaz para matar larvas y pupas de la plaga, especialmente en suelos no estériles, y causó una reducción del 60 % en la supervivencia de los adultos 50 días después de la emergencia. Estos resultados indican que es posible el uso de estos aislados para el control de B. carambolae, lo que sugiere la posibilidad de utilizar este modo de control biológico como otra alternativa para controlar esta plaga cuarentenaria.

Tobacco Growth Promotion by the Entomopathogenic Fungus, Isaria javanica pf185

Isaria javanica pf185 is an important entomopathogenic fungus with potential for use as an agricultural biocontrol agent. However, the effect of I. javanica pf185 on plant growth is unknown. Enhanced tobacco growth was observed when tobacco roots were exposed to spores, cultures, and fungal cell-free culture supernatants of this fungus. Tobacco seedlings were also exposed to the volatiles of I. javanica pf185 in vitro using I-plates in which the plant and fungus were growing in separate compartments connected only by air space. The length and weight of seedlings, content of leaf chlorophyll, and number of root branches were significantly increased by the fungal volatiles. Heptane, 3-hexanone, 2,4-dimethylhexane, and 2-nonanone were detected, by solid-phase micro-extraction and gas chromatography-mass spectrophotometry, as the key volatile compounds produced by I. javanica pf185. These findings illustrate that I. javanica pf185 can be used to promote plant growth, and also as a biocontrol agent of insect and plant diseases. Further studies are necessary to elucidate the mechanisms by which I. javanica pf185 promotes plant growth.