Abstract The use of fungi and bacteria with pathogenic capacity towards insects is considered a sustainable agroecological alternative for efficient pest control, as they are microorganisms of easy production in industrial scale, formulation and use. Therefore, the objective was to evaluate four entomopathogenic formulations in the control of Brevicoryne brassicae L., a key pest of cabbage (Brassica oleracea L.), in a randomized complete block design with 4 treatments, plus an absolute control and 4 replicates. The variables evaluated were the number of dead and live aphids per plant and the number of infested plants. The commercial formulations Biosafe, BesT-K, Metarrizo and Yurak containing strains of Bacillus subtilis, Bacillus thuringiensis var kurstaki (Btk), Metarhizium anisopliae and Bauveria bassiana respectively, are the ones that were used by spray application to the crop in two seasons: planting, season August-December 2019 and August-December 2021. Among the results, M. anisopliae was recorded with up to 91.11 % efficiency, followed by B. bassiana with 89.50 % and Bs with 79.38 % in the reduction of populations under field conditions. In the laboratory, Btk and M. anisopliae reached 100 % mortality in a period of seven days, so it is concluded that in the medium term, entomopathogenic fungi in the field are more efficient in aphid control; however, in controlled conditions Btk is also efficient.

Resumen El uso de hongos y bacterias con capacidad patogénica hacia los insectos se considera una alternativa agroecológica sostenible de lucha eficiente contra las plagas por ser microorganismos de fácil producción en escala industrial, formulación y uso; entonces, el objetivo ha sido evaluar cuatro formulados entomopatógenos en el control de Brevicoryne brassicae L, plaga clave de la col (Brassica oleracea L.), en diseño de bloques completos al azar de 4 tratamientos, sumado un testigo absoluto y 4 réplicas, siendo las variables evaluadas la cantidad de pulgones muertos y vivos por planta y cantidad de plantas infestadas. Los formulados comerciales Biosafe, BesT-K, Metarrizo y Yurak que contenían cepas de Bacillus subtilis, Bacillus thuringiensis var kurstaki (Btk), Metarhizium anisopliae y Bauveria bassiana, respectivamente, son los que fueron utilizados mediante la aplicación por aspersión al cultivo en dos épocas de siembra, temporada agosto-diciembre 2019 y agosto-diciembre 2021. Entre los resultados se registró al M. anisopliae con hasta 91,11 % de eficiencia, seguida por B. bassiana con 89,50 % y Bs con 79,38 % en la reducción de las poblaciones en condiciones de campo; en laboratorio el Btk y M. anisopliae alcanzaron el 100 % de mortalidad en un lapso de siete días, por lo que se concluye que a mediano plazo los hongos entomopatógenos en campo resultan ser más eficientes en el control del pulgón, sin embargo, en condiciones controladas el Btk también es eficiente.

Resumo O setor florestal brasileiro se destaca pela tecnologia, práticas de manejo florestal, responsabilidade social e ambiental e, principalmente, alta produtividade, mas pragas exóticas podem reduzir isto. O parasitoide Psyllaephagus bliteus (Riek, 1962) (Hymenoptera: Encyrtidae), percevejos predadores e fungos entomopatogênicos como Beauveria bassiana e Metarhizium anisopliae são inimigos naturais usados no controle biológico do psilídeo-de-concha do eucalipto Glycaspis brimblecombei (Moore, 1964) (Hemiptera: Aphalaridae), praga em plantios de eucalipto. A utilização de fungos entomopatogênicos contra pragas exóticas vem aumentando no setor florestal e a prospecção e identificação de isolados fúngicos é importante para o manejo integrado de pragas. O objetivo deste estudo foi o isolamento e identificação molecular de Cordyceps spp. e avaliar a patogenicidade dos isolados de Cordyceps javanica (LCBPF 11) e Cordyceps fumosorosea (LCBPF 12 e LCBPF 63), obtidos de adultos de Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae), ao psilídeo-de-concha G. brimblecombei. Os fungos foram isolados de adultos de B. tabaci encontrados em culturas de soja e tomate e identificados molecularmente. Os conídios foram suspensos em solução com Tween 80 (0,1%) na concentração de 1,0 x 108 conídios/mL e pulverizados sobre dez ninfas de G. brimblecombei por folha de Eucalyptus cortada e colocada sobre gel hidrorretentor no interior de placas de Petri por repetição. O número de insetos mortos foi quantificado, diariamente, por sete dias e transferidos para câmaras úmidas. Cordyceps javanica (LCBPF 11) e C. fumosorosea (LCBPF 12 e LCBPF 63) foram identificados por análise molecular e todos os isolados foram patogênicos para os insetos. Isto indica serem importantes para o manejo de G. brimblecombei e confirma relatos de que, normalmente, fungos causam maiores mortalidades em insetos da mesma ordem daquela da qual foram isolados.

Abstract The Brazilian forestry sector stands out for its technology, forestry management practices, social and environmental responsibility and, mainly, for its high productivity and exotic pests can reduce it. The red gum lerp psyllid Glycaspis brimblecombei (Moore, 1964) (Hemiptera: Aphalaridae) is an important pest in Eucalyptus plantations. The parasitoid Psyllaephagus bliteus (Riek, 1962) (Hymenoptera: Encyrtidae), predatory bugs and entomopathogenic fungi such as Beauveria bassiana and Metarhizium anisopliae are the natural enemies and used in the biological control of the red gum lerp psyllid. The use of entomopathogenic fungi against exotic pests is increasing in the forestry sector and the prospecting and identification of fungus isolates is important for integrated pest management. The objective of this work was the isolation and molecular identification of Cordyceps spp. And to evaluate the pathogenicity of isolates, obtained from Bemisia tabaci (Gennadius, 1889) (Hemiptera: Aleyrodidae) adults, against to the red gum lerp psyllid G. brimblecombei. The fungi were isolated from B. tabaci adults found in soybean and tomato crops and molecularly identified. The conidia obtained were suspended in solution with Tween 80 (0.1%) at a concentration of 1.0 × 108 conidia/mL and sprayed on ten G. brimblecombei nymphs per Eucalyptus leaf cut and placed on a hydroretentive gel inside per Petri dishes as a replication. The number of dead insects was quantified, daily, for seven days, and transferred to humid chambers. Cordyceps javanica (LCBPF 11) and C. fumosorosea (LCBPF 12 and LCBPF 63) were identified with a molecular analysis and all isolates were pathogenic to the insects and indicates that they could be used to manage G. brimblecombei and adds to reports that, normally, fungi cause greater mortality on insects of the same order as that from which they were isolated.

Abstract Fusarium oxysporum is an entomopathogenic fungus, and it has anti-biological activity against arthropods. Ticks are blood sucking arthropods which are responsible for transmitting different diseases in humans and animals. The use of chemical insecticides against ticks is not eco-friendly option and results in the development of acaricide resistance. Previously, we had cultured a local isolate of Fusarium oxysporum from soil samples which were identified through microscopy and confirmed through molecular technique. In our previous experiments, we have prepared Silver nanoparticles (AgNP) at pH 7 and they had been characterized through X-Ray Diffraction (XRD), UV-visible and zeta-potential. In our current study, the AgNP were prepared at different pH conditions and characterized through Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). The protein molecules of F. oxysporum were charged with Ag ions. F. oxysporum NP were observed to enhance anti-biological activity by killing Rhipicephalus microplus and they caused 100% mortality at pH 4 and pH 5 in 24 h in anti-tick biological assay. Our study is the first report to do biological assay against Rhipicehalus ticks by using Fusarium AgNP at acidic pH. Biological control using entomopathogenic fungi can be the best alternative of the chemical method to control the tick population.

Resumo Fusarium oxysporum é um fungo entomopatogênico com atividade antibiológica contra artrópodes. Os carrapatos são artrópodes sugadores de sangue responsáveis pela transmissão de diversas doenças em humanos e animais. O uso de inseticidas químicos contra carrapatos não é uma opção ecologicamente correta e resulta no desenvolvimento de resistência acaricida. Anteriormente, havíamos cultivado um isolado local de Fusarium oxysporum a partir de amostras de solo que foram identificadas por microscopia e confirmadas por técnica molecular. Em nossos experimentos anteriores, preparamos nanopartículas de Prata (AgNP) em pH 7 e elas foram caracterizadas por Difração de Raios X (XRD), UV-visível e potencial zeta. No presente estudo, os AgNP foram preparados em diferentes condições de pH e caracterizados através de Microscopia Eletrônica de Varredura (MEV) e Microscopia Eletrônica de Transmissão (TEM). As moléculas de proteína de F. oxysporum foram carregadas com íons Ag. Assim, observou-se que F. oxysporum NP aumenta a atividade antibiológica matando Rhipicephalus microplus e causando 100% de mortalidade em pH 4 e pH 5 em 24h no ensaio biológico anticarrapato. Este estudo é o primeiro relato de caso a realizar um ensaio biológico contra carrapatos Rhipicehalus usando Fusarium AgNP em pH ácido. Nesse sentido, é possível concluir que o controle biológico utilizando fungos entomopatogênicos pode ser a melhor alternativa do método químico para controlar a população de carrapatos.

Abstract Macrofungi, with their extraordinary diversity and ability to adapt to various substrates, play crucial ecological roles in ecosystems. However, their development is strongly influenced by the interaction of several environmental factors. Understanding these factors and their impact on the distribution of fungi is essential to understand their ecological dynamics. To obtain data how these organisms are distributed under certain topographic factors, an analysis of the species richness of macrofungi was carried out in a tropical deciduous forest on the coast of Oaxaca, taking both the eco-nutritional habits and substrate on which, they develop in response to altitude, slope and terrain orientation. A review was conducted of a field database of 251 biological samples representing 169 species of macrofungi registered in the Environmental Management Unit for the Conservation of Wildlife "Jardín Botánico-Puerto Escondido". The review was used to assigned a life form (parasitic, mutualistic micorrhiza, saprotrophic) and substrate preferences (lignicolous, humicolous, terricolous, entomopathogenic parasite). To understand the topographic characteristics of the collection sites, maps of altitude, slope and solar exposure were designed in which the geographical coordinates recorded in the database for each study sample were plotted. The results reveal a predominance of saprotrophic life form in this tropical deciduous forest, with a marked preference for edaphic substrates. In addition, more than 50% of the species analyzed show a tendency to coexist on moderate to gentle slopes, oriented to the north, and at altitudes ranging from 120 to 140 m. This study highlights the complex interactions between the diversity of macroscopic fungi, their nutritional preferences and their relationship with altitude, slope, and terrain orientation. Relevant information is provided that can be used in resource management and conservation strategies, by identifying sites within the study area with the greatest species richness.

Resumen Los hongos macroscópicos, dada su extraordinaria diversidad y capacidad de adaptación a diversos sustratos, desempeñan funciones ecológicas cruciales en los ecosistemas. No obstante, su desarrollo está fuertemente condicionado por la interacción de factores ambientales. La comprensión de esos factores y su impacto en la distribución de los hongos es esencial para entender su dinámica ecológica. Con el fin de conocer cómo se distribuyen estos organismos bajo ciertos factores topográficos, se llevó a cabo un análisis de la riqueza de macromicetos en un bosque tropical caducifolio en la costa de Oaxaca, considerando los hábitos eco-nutricionales y el sustrato donde se desarrollan, en respuesta a la altitud, pendiente y orientación del terreno. El estudio consistió en realizar una revisión de una base de datos de campo de 251 muestras biológicas que representan 169 especies de hongos macroscópicos registradas en la Unidad de Manejo Ambiental para la Conservación de la Vida Silvestre "Jardín Botánico-Puerto Escondido". La revisión permitió determinar la forma de vida (parásita, mutualista micorrícica, saprobia) y las preferencias de sustrato (lignícola, humícola, terrícola, parásito entomopatógeno) de cada uno de los especímenes. Para conocer las características topográficas de los sitios donde fueron recolectados los ejemplares, se diseñaron mapas de altitud, pendiente y exposición solar en los cuales se posicionaron las coordenadas geográficas registradas en la base de datos de cada una de las muestras de estudio. Los resultados revelan un predominio de la forma de vida saprobia en este bosque tropical caducifolio, con una marcada preferencia por sustratos edáficos. Además, más del 50% de las especies analizadas mostraron una tendencia por coexistir en laderas con pendientes medianas y suaves, orientadas hacia el norte y a altitudes que oscilan entre los 120 y 140 m. El estudio resalta las complejas interacciones entre la diversidad de macromicetos, sus preferencias nutricionales en relación con la altitud, pendiente y orientación del terreno. Se provee información relevante que puede ser utilizada en estrategias de manejo y conservación de recursos, ubicando los sitios con mayor riqueza de especies dentro de la zona de estudio.

ABSTRACT This study was conducted in a commercial apple orchard (Malus domestica) with high woolly apple aphid (Eriosoma lanigerum) incidence, which is highly harmful to apple culture and infects the trunk, branch, roots, shoot, and fruit. Although the fungus Beauveria bassiana is widely reported as a control agent, there is no evidence of its control action against this pest. This case study was conducted under natural infestation conditions to evaluate the effective action of B. bassiana in E. lanigerum nymph mortality and aimed to evaluate the control potential of E. lanigerum treated with different formulation doses based on B. bassiana strain ICBBb252. The experimental unit consisted of ten plants in four randomized blocks, where one colony per plant was previously marked in an orchard (2500 plants/ha). The treatments consisted of applying the formulate in 100, 200, and 300 mL/ha dosages at a 109 spore/mL concentration and one control group; 10, 20, 30, and 40 days after application, the treatments were evaluated by counting the number of colonies alive. The control of woolly apple aphids by B. bassiana strain ICBBb252 was more effective at 200 and 300 mL/ha dosages and has no phytotoxic effects on plants.

Abstract: In the present contribution, the survey of entomopathogenic fungal diversity associated with organic yerba mate (Ilex paraguariensis A. St.-Hil.) crops allowed the identification of fungi with bioinsecticidal potential of great importance for pest management in organic crops. For this purpose, samples collected from 12 georeferenced sites of certified organic yerba mate plantations in the district of Bella Vista, Itapúa, were analyzed. Isolation and characterization of all fungal strains found in samples of live and dead insects was performed. The direct seeding plate methodology was applied and the colonies were analyzed macro and microscopically. As a result, and with favorable environmental conditions, it was possible to isolate the entomopathogenic fungus Beauveria as well as the genera Alternaria, Rhizopus, Absidia, Mucor, Aspergillus and Fusarium infecting dead insect tissues. In all sampling sites, Alternaria, Aspergillus and Fusarium genera were abundant and were preserved in straparia for future pathogenicity tests. The low frequency in which entomopathogenic fungi were found could be due to the intense droughts that occurred during the months of December/2021 to February/2022, drastically affecting the presence of insect pests and fungi sensitive to changes in temperature and humidity.

Resumen: En la presente contribución, el relevamiento de la diversidad fúngica entomopatógena asociada a cultivos orgánicos de yerba mate (Ilex paraguariensis A. St.-Hil.) permitió identificar los hongos con potencial bioinsecticida de gran importancia para el manejo de plagas en cultivos orgánicos. Para ello se analizaron muestras colectadas de 12 sitios georreferenciados de los yerbales orgánicos certificados del distrito de Bella Vista, Itapúa. Se realizó el aislamiento y caracterización de todas las cepas fúngicas encontradas en muestras de insectos vivos y muertos. Se aplicó la metodología de siembra directa en placa y se analizaron las colonias macro y microscópicamente. Como resultado, y con las condiciones ambientales favorables se pudo aislar el hongo entomopatógeno Beauveria como así también los géneros Alternaria, Rhizopus, Absidia, Mucor, Aspergillus y Fusarium infectando los tejidos de insectos muertos. En todos los sitios de muestreo abundaron los géneros Alternaria, Aspergillus y Fusarium que fueron conservados en cepario para futuros ensayos de patogenicidad. La baja frecuencia en que se encontraron los hongos entomopatógenos pudo deberse a las intensas sequías que acontecieron durante los meses de diciembre/2021 a febrero/2022, afectando drásticamente la presencia de insectos plagas y la de hongos sensibles a los cambios de temperatura y humedad.

Resumen Las tucuras causan graves pérdidas económicas en la agricultura a nivel mundial. En la actualidad, los insecticidas químicos siguen siendo el único medio utilizado para el control de acridios, pero los efectos de su utilización son ambientalmente preocupantes. El objetivo de este trabajo fue probar la eficacia de la cepa Beauveria bassiana (LPSc 1067) sobre seis especies de tucuras consideradas plagas de Argentina. En cuanto a la patogenicidad de B. bassiana sobre ninfas de tercer estadio de las diferentes especies tratadas, se encontraron diferencias significativas (DF= 5; F= 9.93; P<0.0001). Los valores de mortalidad más altos (100%) se registraron en ninfas de Trimerotropis pallidipennis y Dichroplus maculipennis y la mortalidad más baja se observó en ninfas de Scotussa lemniscata con una mortalidad de 48,6 +3,5%. El tiempo medio de supervivencia (MST) más bajo se registró para T. pallidipennis (3,5 +0,15 días) y el MST más alto se observó en ninfas de Dichroplus pratensis (7,48 +0,28 días). Los resultados sugieren que B. bassiana LPSc 1067 constituye un excelente candidato para ser estudiado en profundidad como agente de control biológico de T. pallidipennis y D. maculipennis.

Abstract Grasshoppers affect agriculture worldwide, causing serious economic damage. Currently, the application of chemical insecticides against grasshoppers is the only effective strategy, even considering the significant environmental concern. This study aimed to test the entomopathogenic fungi Beauveria bassiana (LPSc 1067) as biocontrol agent on six harmful grasshopper species in Argentina. Significant differences were observed (DF= 5; F= 9.93; P<0.0001) when considering B. bassiana pathogenicity on third-instar nymphs of the different grasshopper species. The highest mortality (100%) was registered on Trimerotropis pallidipennis and Dichroplus maculipennis nymphs while the lowest mortality (48.6 ±3.5%) was observed on Scotussa lemniscata nymphs. The lowest mean survival time (MST) was recorded for T. pallidipennis (3.5 ±0.15 days) and the highest MST was observed on Dichroplus pratensis nymphs (7.48 ±0.28 days). Results suggest that B. bassiana LPSc 1067 may constitute an excellent candidate to be further studied as biological control agent of T. pallidipennis and D. maculipennis.

Resumen Sitophilus granarius, es un pequeño insecto el cual bajo condiciones favorable llega a afectar al trigo almacenado hasta en un 85 %. Los esfuerzos investigativos están siendo orientados hacia el desarrollo de bioproductos a base de hongos o bacterias con acción entomopatógena, con utilidad potencial como bioinsecticidas. Estudios relacionados con Trichoderma harzianum y espinosina en el control del coleópteros-plaga de granos almacenados son mínimos y es en este contexto, el objetivo del presente trabajo consistió en evaluar el efecto de Trichoderma y la espinosina en el control de Sitophilus granarius en trigo almacenado. Para el desarrollo del estudio, el grano de todos los tratamientos fue impregnado primeramente con espinosina; tres concentraciones (tres tratamientos) de conidias de T. harzianum fueron asperjadas (T1:103, T2:106 y T3:109 conidias.mL-1), con cinco repeticiones cada tratamiento. En el estudio fueron considerados controles sin tratamientos. Los resultados muestran que, Trichoderma harzianum presenta un efecto biorregulador sobre S. granarius, el cual es significativo cuando se combina con la espinosina. Este efecto bioregulador se enfatiza cuando se inoculan concentraciones superiores de 109 condias/mL. Se deben realizar estudios relacionados con la coinoculación y el uso de la espinosina, asimismo, evaluar la viabilidad de la semilla y propiedades organolépticos del grano.

Abstract Sitophilus granarius, is a small insect which, under favorable conditions, affects up to 85 % of stored wheat. Research efforts are being directed towards the development of bioproducts based on fungi or bacteria with entomopathogenic action, with potential utility as bioinsecticides. Studies related to Trichoderma harzianum and spinosyn in the control of the beetle pest of stored grains are minimal and it is in this context, that the objective of this work was to evaluate the effect of Trichoderma and spinosyn in the control of Sitophilus granarius in stored wheat. For the development of the study, the grain of all treatments was first impregnated with spinosyn; three concentrations (three treatments) of T. harzianum conidia were sprayed (T1:103, T2:106 y T3:109 conidias.mL-1), with five repetitions for each treatment. In the study they were considered controls without treatments. The results show that Trichoderma harzianum has a bioregulatory effect on S. granarius, which is significant when combined with spinosyn. This bioregulatory effect is emphasized when higher concentrations than 109 condia/mL are inoculated. Studies related to coinoculation and the use of spinosyn should be carried out, as well as evaluating the viability of the seed and the organoleptic properties of the grain.

Resumen Macrodactylus nigripes (Bates) es una plaga que ataca el cultivo del maíz causando pérdidas económicas en la producción. Con la finalidad de encontrar alternativas agroecológicas para su manejo, el objetivo fue evaluar el efecto de la aplicación de Argemone mexicana L., Ricinus communis L, Beauveria bassiana (Bals) Vuill y Metharrizium anisoplae (Metschnikoff) Sorokin, así como colecta de M. nigripes con red de golpeo en el cultivo de maíz. El experimento se realizó bajo un diseño de bloques completamente al azar con cuatro repeticiones, donde se probaron los cinco métodos de control y fueron comparados con el tratamiento testigo (agua). R. communis y A. mexicana disminuyeron significativamente la infestación de plantas por M. nigripes, aumentando la producción en 36 y 30% respectivamente. Estos resultados demuestran que ambos tratamientos alternados con jabón zote pueden ser una buena alternativa del manejo de esta especie plaga.

Abstract Macrodactylus nigripes (Bates) is a pest that attacks corn crops, causing economic losses in production. In order to find agroecological alternatives for its management, the objective was to evaluate the effect of the application of Argemone mexicana L., Ricinus communis L, Beauveria bassiana (Bals) Vuill and Metharrizium anisoplae (Metschnikoff) Sorokin, as well as the collection of M. nigripes with a beating net in corn crops. The experiment was conducted under a completely randomized block design with four repetitions, where the five control methods were tested and compared with the control treatment (water). R. communis and A. mexicana significantly decreased plant infestation by M. nigripes, increasing production by 36% and 30%, respectively. These results show that both treatments alternated with zote soap can be a good alternative for the management of this pest species.

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.

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.

Eriosoma lanigerum (Hausmann) (Hemiptera: Aphididae), woolly apple aphid (WAA) is an important pest on apples globally and a key pest of apple production in South Africa. The aphid has developed some level of resistance against several chemical insecticides. Entomopathogenic fungi (EPF) have been identified as promising biological control agents against a wide array of insect pests. The main aim of this study was to conduct a survey in local apple orchards in the Western Cape province for EPF and to use isolates to test the susceptibility of WAA under optimum laboratory conditions. Soil samples were collected from apple orchards and baited using larvae of Galleria mellonella and Tenebrio molitor to isolate EPF. Six EPF species: Beauveria bassiana, Cordyceps fumosorosea, Metarhizium brunneum, M. pinghaense, M. robertsii and Purpureocillium lilacinum were identified from the soil samples. The results from bioassays showed that Metarhizium robertsii and M. pinghaense isolates were the most effective with an average percentage mortality of > 90%. Metarhizium brunneum also proved to be effective when used against the insect with an average percentage mortality of > 80%, while B. bassiana and C. fumosorosea were the least effective with average percentage mortality of 52% and 48%, respectively. The lethal time required to kill 50% and 90% of the colony at a concentration of 107 conidia/ml over five days, LT50 and LT90, of M. robertsii (2.12 and 4.19 days, respectively) and M. pinghaense (2.05 and 4.45 days, respectively) showed to require similar mortality time in days of E. lanigerum. The results obtained in the study have provided an insight into the diversity of EPF species across apple orchards of the Western Cape and shown the efficacy of the Metarhizium isolates as potential biological control agents of the E. lanigerum.

Natural enemies are major challenges in laboratory rearing of grasshoppers, but the identity and virulence of these towards the edible long-horned grasshopper Ruspolia differens (Serville) is scarcely known. In this study, fungi and bacteria were isolated from R. differens collected from Mbarara, Masaka, Hoima, Kampala and Kabale districts in Uganda in 2018, cultured on standard microbial media, identified using molecular techniques and screened for virulence against the insect in laboratory bioassays. Fourteen and nine species of fungi and bacteria were isolated from R. differens, respectively, with the number of isolates varying based on collection site. The most prevalent entomopathogenic fungal species were Aspergillus flavus Link (27.3%), Fusarium equiseti (Corda) (24.2%), Mucorfragilis Fresen (12.1%), Clonostachys rosea (Link) (6.0%) and Aspergillus tamarii Kita (6.0%); whereas the most prevalent bacterial isolates were Serratia marcescens Bizio (38.1(0)%), Bacillus thuringiensis (Berliner) (14.3%) and Enterobacter cloacae (Jordan) (14.3%). Nine of the fungal species namely Clavispora lusitaniae Rodrigues de Miranda, Lichtheimia corymbifera (Cohn), Trichoderma koningii Oudem, F. equiseti, M. fragilis, Aspergillus niger van Tieghem, Epicoccum sorghinum (Saccardo), C. rosea, Penicillium commune Charles Thorn; and five bacterial species (Proteus penneri Hickman, S. marcescens, B. thuringiensis, Staphylococcus sciuri Kloos and Enterococcus faecalis (Andrewes and Horder)) were ~5-7-fold and ~4-5-fold, more lethal to third instars of R. differens than untreated controls, respectively. This study is the first to report C. lusitaniae, Exserohilum mcginnis Padhye and Ajello, E. sorghinum, P. penneri and E. cloacae as insect pathogens. The results suggest a need to quarantine field collected R. differens before introducing them into the insectary, as well as performing antimicrobial practices during rearing of the insect to prevent entomopathogen-based mortality.

Formulation of entomopathogens refers to the mixing of various inert ingredients, like clays and mineral oils, with the active ingredients which are the entomopathogens. Successful formulation enhances the survival of the entomopathogen and eases their transportation, storage, preparation and application. The aim of this study was to develop a formulation to maintain the longevity and pathogenicity of the mass-produced conidia of local Metarhizium pinghaense and M. robertsii, for above-ground future commercial field application against Pseudococcus viburni. The objectives were to develop a cost-effective protocol for formulation of infective propagules and to test their effectiveness under laboratory conditions. The conidia of both isolates were formulated using four different oils (liquid paraffin, coconut, canola and olive oils) as liquid carriers, and diatomaceous earth as a mineral carrier. Conidial viability and pathogenicity were assessed over a period of eight weeks. Conidia formulated in oil carriers maintained a high conidial viability and survival rate of > 95% over a period of eight weeks for both isolates, relative to when formulated in mineral carriers, or when stored as dry conidial powder. The conidia in all the oil formulations were also observed to induce high mortality, ranging between 60% and 90% for M. pinghaense, and between 70% and 90% for M. robertsii, when used against P. viburni. The ability of conidia of both isolates to maintain viability and pathogenicity, following storage in the oil formulations, increased the likelihood of the local isolates being successfully integrated as biological control agents for management of P. viburni under field conditions.

Entomopathogenic fungi (EPF) are cosmopolitan soil borne pathogens that cause epizootics in various insect orders. EPF isolates of Metarhizium brunneum and M. pinghaense have shown the potential for use as biological agents of important agricultural insect pests. The aim of the current study was to test for the persistence of M. brunneum and M. pinghaense on apple bark over a period of three weeks, under laboratory conditions. Apple bark was sprayed with conidial suspensions of both fungi, at a standard infective conidial concentration of 10(7) conidia/ml. The persistence, or survival, of the conidia on apple bark was measured using codling moth larvae (CM) (Cydia pomonella) and females of woolly apple aphid (WAA) (Eriosoma lanigerum) as indicator species. The results showed that conidia of M. pinghaense can induce mortality of insect pests through contact with an EPF-treated substrate, with mortality of 39% to 82% for WAA over a period of 10 days post application, and with mortality of 3% to 68% for CM over a period of 7 days, after application to apple bark. Further evaluation showed that the conidia of M. pinghaense persisted longer on apple bark, up to 63%, than did M. brunneum, up to 11%, three weeks post application of the conidial suspensions. The study provides insights into the potential persistence of fungal isolates on apple bark over time post application. Further evaluation of the persistence of the isolates on apple bark under both glasshouse and field conditions should be conducted.

Entomopathogenic fungi (EPF) are effective and environment-friendly insect biological control agents. Ultraviolet (UV) light is known to have an effect on the survival of fungal conidia, and natural sunlight is potentially one of the most damaging factors undermining EPF persistence and pathogenicity. This study aimed to test the infection potential of an isolate of Beauveria bassiana and five Metarhizium species after exposure to different light treatments, on soil and leaf surfaces under laboratory and field conditions, using Thaumatotibia leucotreta (Lepidoptera: Tortricidae) as the test host. Conidia were exposed either to growth light alone, which emits the same visible light as the sun, but excluding UV light, or directly exposed to UV light for 12 h. The results indicated no negative effect on the infection potential of the conidia of most species tested. The conidia of the two Metarhizium pinghaense (5HEID and TH149) isolates showed the greatest tolerance to visible light and UV radiation exposure on both soil and leaf surfaces. Exposure of M. pinghaense isolates to visible light on soil surfaces showed pathogenicity of > 80% for both isolates, and of between 58% and 88% after exposure to UV light. On leaf surfaces, three Metarhizium isolates, M. pinghaense (5HEID and TH149) and M. majus (TH153) had > 90% pathogenicity following exposure to UV light, and M. pinghaense (TH149) and M. robertsii (6EIKEN) showed greater tolerance of > 70%, under laboratory conditions. However, the pathogenicity of the EPF isolates was very low in field trials, indicating that further trials on the use of formulations and adjuvants with the isolates are needed to improve long-term persistence and efficacy under field conditions.