Purpureocillium lilacinum SBF054: Endophytic in Phaseolus vulgaris, Glycine max, and Helianthus annuus; Antagonistic to Rhizoctonia solani; and Virulent to Euschistus heros.

Microorganisms with multiple ecological functions can be a useful biotechnological resource in integrated pest- and disease-management programs. This work aimed to investigate the potential endophytic and virulent effects of a strain of Purpureocillium lilacinum on organic cultivation in Brazil. Specifically, the strain's ability to establish itself as an endophyte in common bean, soybean, and sunflower plants when inoculated via seed was evaluated. Furthermore, its antifungal activity against phytopathogens and its pathogenicity and virulence against insects of the order Lepidoptera, Coleoptera, and Hemiptera were evaluated. Furthermore, the strain was evaluated for its biochemical and physiological characteristics. For virulence bioassays, the experiments were conducted under a factorial scheme (2 × 3), with the following factors: (a) fungal inoculation and control without inoculum and (b) types of inocula (blastospores, aerial conidia, and metabolites). The treatments were sprayed on insect species at different stages of development. In summary, it was found that the SBF054 strain endophytically colonized the common bean, with partial recovery from the root tissues of soybean and sunflower plants, 30 days after inoculation; suppressed 86% of Rhizoctonia solani mycelial growth in an in vitro assay; and controlled eggs, nymphs, and Euschistus heros adults. These multifunctional abilities are mainly attributed to the strain's mechanisms of producing metabolites, such as organic acids, soluble nutrients, and hydrolytic enzymes.

Fusarium oxysporum; its enhanced entomopathogenic activity with acidic silver nanoparticles against Rhipicephalus microplus ticks / Fusarium oxysporum e sua atividade entomopatogênica aprimorada com nanopartículas de prata ácida contra carrapatos Rhipicephalus microplus

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.

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.

Impact of osmotic stress on production, morphology, and fitness of Beauveria bassiana blastospores.

Beauveria bassiana is a cosmopolitan entomopathogenic fungus that can infect over 1000 insect species. During growth inside the host, B. bassiana transitions from hyphal to yeast-like unicellular growth as blastospores. Blastospores are well suited as an active ingredient in biopesticides due to their ease of production by liquid fermentation. Herein, we investigated the impact of hyperosmotic growth environments mediated by ionic and non-ionic osmolytes on two strains of B. bassiana (ESALQ1432 and GHA) relevant to growth morphology, blastospore production, desiccation tolerance, and insecticidal activity. Polyethylene glycol (PEG200) increased osmotic pressure in submerged cultures leading to decreased blastospore size but higher blastospore yields for one strain. Morphologically, decreased blastospore size was linked to increased osmotic pressure. However, smaller blastospores from PEG200 supplemented cultures after air-drying exhibited delayed germination. Ionic osmolytes (NaCl and KCl) generated the same osmotic pressure (2.5-2.7 MPa) as 20% glucose and boosted blastospore yields (> 2.0 × 109 blastospores mL-1). Fermentation performed in a bench-scale bioreactor consistently promoted high blastospore yields when using NaCl (2.5 MPa) amended media within 3 days. Mealworm larvae (Tenebrio molitor) were similarly susceptible to NaCl-grown blastospores and aerial conidia in a dose-time-dependent manner. Collectively, these results demonstrate the use of hyperosmotic liquid culture media in triggering enhanced yeast-like growth by B. bassiana. Understanding the role of osmotic pressure on blastospore formation and fitness will hasten the development of viable commercial fungal biopesticides. KEY POINTS: • Osmotic pressure plays a critical role in submerged fermentation of B. bassiana. • Ionic/non-ionic osmolytes greatly impact blastospore morphology, fitness, and yield. • Desiccation tolerance and bioefficacy of blastospores are affected by the osmolyte.

Biocontrol of Xyleborus affinis (Curculionidae: Scolitinae) Females and Progeny by Beauveria bassiana (Hypocreales: Cordycipitaceae) in a Sawdust Artificial Diet Model.

The ambrosia beetle Xyleborus affinis, recently reported affecting avocado trees in Mexico, represents one of the most widespread insects worldwide. Previous reports have shown that Xyleborus genera members are susceptible to Beauveria bassiana and other entomopathogenic fungus strains. However, their effect on borer beetles' progeny has not been fully investigated. The aim of the present study was to determine the insecticidal activity of B. bassiana on X. affinis adult females and their progeny in an artificial sawdust diet bioassay model. The B. bassiana strains CHE-CNRCB 44, 171, 431, and 485 were individually tested on females at concentrations ranging from 2 × 106 to 1 × 109 conidia mL-1. After 10 d of incubation, diet was evaluated to count laid eggs, larvae, and adults. Insect conidia loss after exposure was determined by attached conidia to each insect after 12 h of exposure. The results showed that females' mortality ranged between 3.4% and 50.3% in a concentration-response manner. Furthermore, we did not observe statistical differences among strains at the highest concentration. CHE-CNRCB 44 showed the highest mortality at the lowest concentration and reduced larvae and laid eggs at the highest concentration (p < 0.01). Strains CHE-CNRCB 44, 431, and 485 significantly decreased larvae, as compared with the untreated control. After 12 h, up to 70% of conidia was removed by the effect of the artificial diet. In conclusion, B. bassiana has the potential to control X. affinis adult females and progeny.

Study on the insecticidal activity of entomopathogenic fungi for the control of the fruit fly (Anastrepha obliqua), the main pest in mango crop in Colombia.

The aim of this study was to evaluate and select entomopathogenic fungi that produces insecticidal compounds for the control of adults of Anastrepha obliqua Macquart (Diptera: tephritidae) that are the main pest of mango (Mangifera indica L. Bark) in Colombia. Nine entomopathogenic fungi isolates were evaluated, five belonging to the genus Metarhizium and four belonging to the genus Beauveria. One strain of the species Metarhizium robertsii with insecticidal activity was selected. By column fractionation, an active fraction was obtained, which caused mortalities higher than 90% after 48 h of exposure. Through HPLC it was determined that the active fraction is composed of more than 22 metabolites. Identification of the metabolites by UHPLC MS/MS revealed the presence of destruxin in E, D, A and B groups (destruxin E-diol, destruxin D, destruxin D1, destruxin D2, destruxin A2, destruxin A, destruxin A3, dihydrodestruxin A, desmB, destruxin B2, destruxin B and destruxin B1). The evaluation of the insecticidal capacity of the organic fractions obtained by HPLC indicated that the extract obtained from the isolate M. robertsii had a compound with high activity on adults of A. obliqua (destruxin A) causing massive mortality of up to 100%, after 48 h of the treatment administration. Furthermore, two other compounds with medium activity were found (destruxin A2 and destruxin B), showing mortalities between 60.0 and 81.3%, respectively. The extract of the isolate MT008 of M. robertsii showed higher insecticidal activity and a potential source for the control of A. obliqua.

Natural infection of Chiromyzinae larvae (Diptera: Stratiomyidae) in southern Chile by Tolypocladium valdiviae sp. nov.

A new species from the fungal genus Tolypocladium (Hypocreales: Ophiocordycipitaceae) that infects Stratiomyidae larva from the genus Hylorops is described: Tolypocladium valdiviae Gallardo-Pillancari, Montalva & González. The description is based on both genomic data and morphological characteristics. The sexual stage of T. valdiviae presents fleshy and visible stromata; unlike Tolypocladium ophioglossoides, it is smaller and emerges directly from its host and resembles Tolypocladium longisegmentis and Tolypocladium capitatum, both of which are parasites of deer truffle fungi of the genus Elaphomyces (Ascomycota: Eurotiales). In the anamorphic state, T. valdiviae presents conidiogenous cells similar in shape and arrangement to those of Tolypocladium inflatum, however T. valdiviae produces larger conidiogenous cells and, occasionally, produces chlamydospores. Phylogenetic evidence suggested that T. valdiviae is in a clade close to T. longisegmentis, T. inflatum and T. ophioglossoides, species also recognized to be parasites of fungi of the genus Elaphomyces. The new species is known so far only from Valdivia, southern Chile.

In Planta Detection of Beauveria bassiana (Ascomycota: Hypocreales) Strains as Endophytes in Bean (Phaseolus vulgaris L.).

Beauveria bassiana (B. bassiana) is a significant entomopathogenic fungus (EPF) in agriculture as a sprayable biocontrol agent. It has the potential to be established as an endophyte (ENP) in various crops, resulting in beneficial effects for the host plants, including resistance to pest insects and increased growth and yield. However, it is not known whether a B. bassiana strain has such a favorable impact on the plant, since it is a common soil microorganism. Therefore, techniques that allow strain monitoring will be advantageous. To date, methods for detecting or monitoring a specific EPF strain after external application are scarce. In the present study, an in planta nested PCR technique was standardized to differentiate between three B. bassiana strains (GHA, PTG4, and BB37) established as endophytes in bean plants under laboratory conditions by detecting the insertion profile of four group I introns located in the 28S gene of B. bassiana ribosomal DNA. This technique recognized a distinct pattern of bands of different sizes for each strain, with a sensitivity of 1 pg per 10 ng of plant DNA. This molecular approach may be more effective monitoring B. bassiana strains after application to evaluate their significance on crops.

Clonostachys spp., natural mosquito antagonists, and their prospects for biological control of Aedes aegypti.

Aedes aegypti (Linnaeus, 1762) is an important vector of arboviruses in the tropics and subtropics. New control strategies based on natural enemies such as entomopathogenic fungi are of utmost importance, and the present study reports the first isolation of Clonostachys spp. (Hypocreales: Bionectriaceae) from mosquitoes and their activity against A. aegypti. Entomopathogenic fungi were surveyed in central Brazil using A. aegypti larvae as sentinels and, also, a CDC light trap. Clonostachys eriocamporesii R.H. Perera & K.D. Hyde, 2020 (IP 440) and Clonostachys byssicola Schroers, 2001 (IP 461) were identified by sequence analysis of the nuclear ribosomal internal transcribed spacer gene, and tested against eggs, larvae, and adults. Both strains were highly active against A. aegypti third instar larvae, with mortalities ≥ 80% at 107 conidia/mL after 5 days but distinctly less active against eggs and adults. This is the first report of both C. eriocamporesii and C. byssicola as naturally occurring pathogens affecting mosquitoes, and IP 440 appears to be a promising control agent against aquatic stages of A. aegypti.

Occurrence of entomopathogenic hypocrealean fungi in mosquitoes and their larval habitats in Central Brazil, and activity against Aedes aegypti.

Collecting entomopathogenic fungi associated with mosquitoes and studies on their activity against mosquito developmental stages will improve the understanding of their potential as agents to control important mosquito vectors. Twenty-one strains of entomopathogenic fungi affecting mosquitoes in Central Brazil were studied: 7 of Beauveria bassiana, 7 of Metarhizium humberi, 3 of M. anisopliae, 2 of Cordyceps sp. and one each of Akanthomyces saksenae and Simplicillium lamellicola. These fungi were isolated from field-collected mosquito adults (3 strains) or larvae (a single strain); the other 17 strains were isolated from laboratory-reared Aedes aegypti sentinel larvae set out in partially immersed cages placed in diverse small- to middle-sized aquatic mosquito habitats in or close to areas with secondary tropical forest. The frequent recovery of normally soil-borne Metarhizium spp. and B. bassiana from aquatic habitats is notable. Our laboratory findings indicated that M. anisopliae IP 429 and IP 438 and M. humberi IP 421 and IP 478 were highly active against immature stages and, together with M. anisopliae IP 432, also against adults. These strains appear to be the most promising candidates to develop effective control strategies targeting the different developmental stages of A. aegypti, the most important vector of viral diseases in humans in the tropics.

Side effects of a fungus-based biopesticide on stingless bee guarding behaviour.

Pathogenic fungi have been used worldwide to control crop pests and are assumed to pose negligible threats to the survival of pollinators. Although eusocial stingless bees provide essential pollination services and might be exposed to these biopesticides in tropical agroecosystems, there is a substantial knowledge gap regarding the side effects of fungal pathogens on behavioural traits that are crucial for colony functioning, such as guarding behaviour. Here, we evaluated the effect of Beauveria bassiana on the sophisticated kin recognition system of Tetragonisca angustula, a bee with morphologically specialized entrance guards. By combining behavioural assays and chemical analyses, we show that guards detect pathogen-exposed nestmates, preventing them from accessing nests. Furthermore, cuticular profiles of pathogen-exposed foragers contained significantly lower amounts of linear alkanes than the unexposed ones. Such chemical cues associated with fungal conidia may potentially trigger aggression towards pathogen-exposed bees, preventing pathogen spread into and among colonies. This is the first demonstration that this highly abundant native bee seems to respond in a much more adaptive way to a potentially infectious threat, outweighing the costs of losing foraging workforce when reducing the chances of fungal pathogen outbreaks within their colonies, than honeybees do.

Genomic signatures and insights into host niche adaptation of the entomopathogenic fungus Metarhizium humberi.

The genus Metarhizium is composed of species used in biological control programs of agricultural pests worldwide. This genus includes common fungal pathogen of many insects and mites and endophytes that can increase plant growth. Metarhizium humberi was recently described as a new species. This species is highly virulent against some insect pests and promotes growth in sugarcane, strawberry, and soybean crops. In this study, we sequenced the genome of M. humberi, isolate ESALQ1638, and performed a functional analysis to determine its genomic signatures and highlight the genes and biological processes associated with its lifestyle. The genome annotation predicted 10633 genes in M. humberi, of which 92.0% are assigned putative functions, and ∼17% of the genome was annotated as repetitive sequences. We found that 18.5% of the M. humberi genome is similar to experimentally validated proteins associated with pathogen-host interaction. Compared to the genomes of eight Metarhizium species, the M. humberi ESALQ1638 genome revealed some unique traits that stood out, e.g., more genes functionally annotated as polyketide synthases (PKSs), overrepresended GO-terms associated to transport of ions, organic and amino acid, a higher percentage of repetitive elements, and higher levels of RIP-induced point mutations. The M. humberi genome will serve as a resource for promoting studies on genome structure and evolution that can contribute to research on biological control and plant biostimulation. Thus, the genomic data supported the broad host range of this species within the generalist PARB clade and suggested that M. humberi ESALQ1638 might be particularly good at producing secondary metabolites and might be more efficient in transporting amino acids and organic compounds.

Low- or high-white light irradiance induces similar conidial stress tolerance in Metarhizium robertsii.

White light during mycelial growth influences high conidial stress tolerance of the insect-pathogenic fungus Metarhizium robertsii, but little is known if low- or high-white light irradiances induce different stress tolerances. The fungus was grown either in the dark using two culture media: on minimal medium (Czapek medium without sucrose = MM) or on potato dextrose agar (PDA) or PDA medium under five different continuous white light irradiances. The stress tolerances of conidia produced on all treatments were evaluated by conidial germination on PDA supplemented with KCl for osmotic stress or on PDA supplemented with menadione for oxidative stress. Conidia produced on MM in the dark were more tolerant to osmotic and oxidative stress than conidia produced on PDA in the dark or under the light. For osmotic stress, growth under the lower to higher irradiances produced conidia with similar tolerances but more tolerant than conidia produced in the dark. For oxidative stress, conidia produced under the white light irradiances were generally more tolerant to menadione than conidia produced in the dark. Moreover, conidia produced in the dark germinated at the same speed when incubated in the dark or under lower irradiance treatment. However, at higher irradiance, conidial germination was delayed compared to germination in the dark, which germinated faster. Therefore, growth under light from low to high irradiances induces similar conidial higher stress tolerances; however, higher white light irradiances cause a delay in germination speed.

Optimization of granular formulations of Metarhizium humberi microsclerotia with humectants.

Granular microsclerotial formulations of entomopathogenic fungi deserve attention because of their post-application, in situ production of new conidia that enhance and prolong mycoinsecticidal efficacy against a target pest insect. Because high ambient moisture is a crucial condition to induce fungal development and conidiogenesis on granules, we tested the impacts of the additions of three humectants-glycerin, propylene glycol, and polyethylene glycol 400-on water absorption by pellets incorporating microsclerotia of Metarhizium humberi IP 46 with microcrystalline cellulose or vermiculite carriers, and on the production of infective conidia of IP 46 microsclerotia in ambient humidities suboptimal for routine conidiogenesis. Glycerin facilitated greater and faster absorption of water than the other humectants. Microcrystalline cellulose absorbed low quantities of water without any added humectant whereas vermiculite did not. IP 46 did not grow or sporulate on pellets prepared with or without glycerin at 86% relative humidity (RH) or on control pellets without glycerin at 91% RH; conidial production on pellets prepared with vermiculite or microcrystalline cellulose and 10% glycerin reached 1.1 × 105 conidia/mg and 1 × 105 conidia/mg, respectively, after 20 days of exposure at 91% RH. Hence, these results strongly support glycerin as a suitable humectant for granular microsclerotial formulations of this fungus.

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. KEY POINTS: • High conidial production on granular microsclerotial formulations at >96.5% RH • Vermiculite is more appropriate as a carrier than microcrystalline cellulose • Granules with IP 46 microsclerotia are highly active against Aedes aegypti adults.

Fumigant activity of essential oils from Cinnamomum and Citrus spp. and pure compounds against Dermanyssus gallinae (De Geer) (Acari: Dermanyssidae) and toxicity toward the nontarget organism Beauveria bassiana (Vuill.).

Dermanyssus gallinae(De Geer) (Acari: Dermanyssidae) is the main ectoparasite associated with laying poultry. This mite is commonly controlled by the application of synthetic chemical insecticides, wich lead to the selection of resistant populations and formation of residues in eggs. Thus, new molecules must be developed to control D. gallinae. This work evaluated the toxicity of essential oils (EOs) from Cinnamomum cassia, Cinnamomum camphora, Cinnamomum camphora var. linalooliferum, Citrus aurantium, Citrus aurantium var. bergamia, Citrus aurantifolia and Citrus reticulata var. tangerine against D. gallinae. Additionally, the chemical profiles of the most bioactive EOs were analyzed by gas chromatography coupled with mass spectrometry (GC-MS) and the major compounds were subjected to new tests using D. gallinae. The most toxic EOs against D. gallinae were evaluated for the nontarget entomopathogenic fungus Beauveria bassiana (Unioeste 88). The EOs from C. cassia (LC50 = 25.43 ± 1.0423 µg/cm3) and C. camphora var. linalooliferum (LC50 = 39.84 ± 1.9635 µg/cm3) were the most active in the fumigant bioassay and caused mortality rates of 96 and 61%, respectively. The GC-MS analysis revealed that the major constituents of EOs from C. cassia and C. camphora var. linalooliferum were trans-cinnamaldehyde and linalool, respectively. The pure compounds, trans-cinnamaldehyde (LC50 = 68.89 ± 3.1391 µg/cm3) and linalool (LC50 = 51.45 ± 1.1967 µg/cm3), were tested on D. gallinae and showed lower toxicity than the EOs. Thus, the compounds were not the only active substances produced by C. cassia and C. camphora var. linalooliferum; moreover synergism may have occurred between the substances. The EOs from C. cassia and C. camphora var. linalooliferum were also toxic to B. bassiana (Unioeste 88). Thus, EOs from C. cassia and C. camphora var. linalooliferum are promising candidates for use in D. gallinae control, but cannot be used in conjunction with the fungus B. bassiana.

Susceptibility of All Nymphal Stages of Bemisia tabaci Biotype B (Hemiptera: Aleyrodidae) to Three Brazilian Isolates of Cordyceps sp. (Hypocreales: Cordycipitaceae) in a Screenhouse Under Variable Temperature and Moisture Conditions.

The susceptibility of 1st to 4th instars of Bemisia tabaci (Gennadius, 1989) (Hemiptera: Aleyrodidae) to three isolates of Cordyceps sp. (Hypocreales: Cordycipitaceae) was evaluated in screenhouse experiments under variable temperatures and moisture conditions. No differences in susceptibility to the Cordyceps sp. isolates were observed among 1st, 2nd, and 3rd instar nymphs with respect to median lethal time (LT50) values. Confirmed mortalities ranged from 63.7 to 87.8% when the isolates were tested at 5 × 107 conidia mL-1. The 4th instar was the least susceptible to the fungal isolates (≤ 36.6% mortality). However, 60.0 to 99.5% of the adults that emerged from 4th instar nymphs previously treated with the fungus succumbed to the infection. Temperature was more detrimental to Cordyceps sp. virulence towards B. tabaci nymphs than relative humidity (RH). At similar RH, median LT50 for 1st instar (9.4 days) was higher than for 3rd instar (5.3 days) when the fungus was tested at 5 × 107 conidia mL-1; minimal temperatures of ≥ 12.6°C compared to ≥ 17.0°C were registered for experiments with 1st and 3rd instars, respectively. However, temperatures ≥ 35°C for 4 to 6 h daily did not affect the efficacy of the fungus against nymphs. Cordyceps sp. showed high virulence to all life stages of B. tabaci at relatively low RH, and an ability to grow extensively over the leaf surface and to produce high amounts of conidia on infected hosts. These attributes certainly boost its potential as an important pest control component of B. tabaci biotype B, especially for management of populations resistant to synthetic insecticides.

UV-B Radiation Tolerance and Temperature-Dependent Activity Within the Entomopathogenic Fungal Genus Metarhizium in Brazil.

Metarhizium comprises a phylogenetically diverse genus of entomopathogenic fungi. In Brazil, Metarhizium anisopliae s.str. subclade Mani 2 is predominantly isolated from insects, while M. robertsii and M. brunneum mostly occur in the soil environment. Solar radiation and high temperatures are important abiotic factors that can be detrimental to fungal propagules. We hypothesized that among 12 Brazilian isolates of Metarhizium spp., M. anisopliae Mani 2 (n = 6), being adapted to abiotic conditions of the phylloplane, is more tolerant to UV light and high temperatures than M. robertsii (n = 3) and M. brunneum (n = 3). Inoculum of each isolate was exposed to UV-B for up to 8 h and viability evaluated 48 h later. After 8 h under UV-B, most of the isolates had germination rates below 5%. Discs of mycelia were incubated at different temperatures, and diameter of colonies were recorded for 12 days. Mycelia of M. robertsii isolates grew faster at 33 °C, while M. anisopliae and M. brunneum grew most at 25 °C. Dry conidia were incubated at 20, 25 or 40 °C for 12 days, and then viabilities were examined. At 40 °C, conidia of five M. anisopliae isolates were the most tolerant. In the three experiments, considerable intra- and inter-specific variability was detected. The results indicate that conclusions about tolerance to these abiotic factors should be made only at the isolate level.

Production of conidia using different culture media modifies the virulence of the entomopathogenic fungus Metarhiziumys against Aedes aegypti larvae.

BACKGROUND & OBJECTIVES: Entomopathogenic fungi are being investigated for the biological control of a range of mosquitoes. Metarhizium conidiospores (conidia) effectively kill Aedes aegypti larvae and could be deployed as an alternative to chemical insecticides. Conidial yield and virulence of fungi when cultured on three different types of solid media, was investigated. METHODS: Three culture media were tested: a) Sabouraud dextrose agar (SDA); b) rice flour yeast agar (RYA) and c) rice grains. Conidia produced using these substrates were tested for virulence against Ae. aegypti larvae obtained from field collected eggs. Larvae (2nd - 3rd instar) were exposed to aqueous conidial suspensions and survival monitored over 7 days. Survival analysis was performed using Log-Rank and Kaplan Meier tests, while fungal growth and conidial yields were analyzed using a two-way ANOVA. RESULTS: There were only small differences between growth rates on RYA and SDA; however, ESALQ 818 showed the highest conidial yield on rice. Conidia produced on rice grains were more virulent, rapidly reducing survival rates of mosquito larvae. ESALQ 818 conidia produced on rice grains, RYA and SDA killed 100% of the larvae on the 2nd, 3rd and 4th day of exposure, respectively. IP 46 virulence of was consistently lower than ESALQ 818 for all the media tested. INTERPRETATION & CONCLUSION: The choice of culture media can influence the virulence of fungal conidia to Ae. aegypti larvae, demonstrating the importance of not only selecting the most virulent isolate but also standardizing growth conditions when screening for virulence.

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

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.

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.

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

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.(AU)

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.(AU)