Community composition of the entomopathogenic fungal genus Metarhizium in soils of tropical and temperate conventional and organic strawberry fields.

Studies on community composition and population structure of entomopathogenic fungi are imperative to link ecosystem functions to conservation biological control. We studied the diversity and abundance of Metarhizium spp. from soil of conventionally and organically farmed strawberry crops and from the adjacent field margins in two different climatic zones: Brazil (tropical) and Denmark (temperate), using the same isolating methods. In Brazilian strawberry soil, Metarhizium robertsii (n = 129 isolates) was the most abundant species, followed by M. humberi (n = 16); M. anisopliae (n = 6); one new taxonomically unassigned lineage Metarhizium sp. indet. 5 (n = 4); M. pingshaense (n = 1) and M. brunneum (n = 1). In Denmark, species composition was very different, with M. brunneum (n = 33) being isolated most commonly, followed by M. flavoviride (n = 6) and M. pemphigi (n = 5), described for the first time in Denmark. In total, 17 haplotypes were determined based on MzFG543igs sequences, four representing Danish isolates and 13 representing Brazilian isolates. No overall difference between the two climatic regimes was detected regarding the abundance of Metarhizium spp. in the soil in strawberry fields and the field margins. However, we found a higher Shannon's diversity index in organically managed soils, confirming a more diverse Metarhizium community than in soils of conventionally managed agroecosystems in both countries. These findings contribute to the knowledge of the indigenous diversity of Metarhizium in agricultural field margins with the potential to contribute to pest regulation in strawberry cropping systems.

Bioactivity Screening and Chemical Characterization of Biocompound from Endophytic Neofusicoccum parvum and Buergenerula spartinae Isolated from Mangrove Ecosystem.

The discovery of biomolecules has been the subject of extensive research for several years due to their potential to combat harmful pathogens that can lead to environmental contamination and infections in both humans and animals. This study aimed to identify the chemical profile of endophytic fungi, namely Neofusicoccum parvum and Buergenerula spartinae, which were isolated from Avecinnia schaueriana and Laguncularia racemosa. We identified several HPLC-MS compounds, including Ethylidene-3,39-biplumbagin, Pestauvicolactone A, Phenylalanine, 2-Isopropylmalic acid, Fusaproliferin, Sespendole, Ansellone, Calanone derivative, Terpestacin, and others. Solid-state fermentation was conducted for 14-21 days, and methanol and dichloromethane extraction were performed to obtain a crude extract. The results of our cytotoxicity assay revealed a CC50 value > 500 µg/mL, while the virucide, Trypanosoma, leishmania, and yeast assay demonstrated no inhibition. Nevertheless, the bacteriostatic assay showed a 98% reduction in Listeria monocytogenes and Escherichia coli. Our findings suggest that these endophytic fungi species with distinct chemical profiles represent a promising niche for further exploring new biomolecules.

Potential Impact of Chemical Fungicides on the Efficacy of Metarhizium rileyi and the Occurrence of Pandora gammae on Caterpillars in Soybean Crops.

Entomopathogenic fungi may play a crucial role in the regulation of caterpillar populations in soybean crops, either through natural occurrences or applied as mycopesticides. In the present work, we reported the naturally occurring entomopathogenic fungus Pandora gammae attacking the caterpillar Chrysodeixis includens, with infection rates in field trials ran in two consecutive years in the 10-35% range. As many chemicals are potentially harmful to entomopathogenic fungi, this work aimed to investigate the potential impact of two chemical fungicides (azoxystrobin + benzovindiflupyr and trifloxistrobina + prothioconazole) used to control soybean rust (Phakopsora pachyrhizi) on the natural occurrence of P. gammae and Metarhizium rileyi, as well as the efficacy of the latter fungus applied as different formulations against the soybean caterpillars Anticarsia gemmatalis and C. includens. Under laboratory conditions, fungicides used at field-recommended rates had a considerable negative impact on the germinability of M. riley on the medium surface, and all tested formulations did not protect conidia from damage by these chemicals. This harmful effect also impacted host infectivity, as the larval mortality owing to this fungus was reduced by 30-40% compared to that of the fungicide-free treatments. In field trials conducted in two subsequent years, unformulated and formulated M. rileyi conidia applied to soybean plants produced primary infection sites in caterpillar populations after a single spray. Spraying unformulated or formulated M. rileyi conidia following fungicide application on plants did not affect host infection rates over time. Moreover, the use of M. rileyi-based formulations or chemical fungicide did not interfere with the natural infection rates by P. gammae on its host, C. includens. Although a higher degree of exposure to non-selective fungicides can negatively affect fungal entomopathogens, a single foliar application of fungicides may be harmless to both M. rileyi and P. gammae in soybean fields. Additionally, this work showed that naturally occurring wasps and tachnids also play an important role in the regulation of A. gemmatalis and, notably, C. includens, with parasitism rates above 40-50% in some cases.

Biological control of Biomphalaria, the intermediate host of Schistosoma spp.: a systematic review / Controle biológico de Biomphalaria, hospedeiro intermediário do Schistosoma spp.: uma revisão sistemática

ABSTRACT: Schistosomiasis is an important vector-borne disease transmitted by an intermediate host: a freshwater mollusk. Control of these snail vectors is one of the strategies of the World Health Organization against the disease. The present study was based on a systematic review of published scientific papers concerning the biological control of snails (genus Biomphalaria), and identified the ongoing challenges and propose future perspectives. The review methodology was based on the PRISMA statement, the international databases Web of Science and Scopus for the period 1945-2021. In total, 47 papers were analyzed, published by authors from 14 different countries, the majority being from: France, Brazil, the United States, and Egypt. The most widely used strategy for biological control was predation by fish (12 studies). Fourteen papers were published in the most prolific decade 2010-2019; during which there was also a greater diversity of biological control agents in studies. In this context, we believed that one of the principal challenges of this approach is the successful simultaneous use of multiple types of biological control agent: predators, competitors, and/or microbial agents. This new approach may provide important insights for the development of new biological control agents or microbial-based products, with the potential to reduce the parasite load carried by schistosomiasis snail vector and control its transmission in a sustainable way.

RESUMO: A esquistossomose é uma importante doença transmitida por vetor, um hospedeiro intermediário: um molusco de água doce. O controle desses caramujos vetores é uma das estratégias da Organização Mundial da Saúde para controle da doença. O presente estudo foi baseado em uma revisão sistemática de artigos científicos publicados sobre o controle biológico de caramujos (gênero Biomphalaria), e teve como objetivo identificar os desafios atuais e propor perspectivas futuras. A metodologia de revisão foi baseada na declaração PRISMA, nas bases de dados internacionais, Web of Science e Scopus, entre 1945-2021. No total, foram analisados 47 artigos, publicados por autores de 14 países diferentes, sendo a maioria: França, Brasil, Estados Unidos e Egito. A estratégia mais utilizada para controle biológico foi a predação por peixes (12 estudos). Quatorze artigos foram publicados na década mais produtiva 2010-2019, durante a qual também houve uma maior diversidade de agentes de controle biológico em estudos. Neste contexto, acreditamos que um dos principais desafios desta abordagem é a utilização simultânea bem-sucedida de múltiplos tipos de agentes de controle biológico: predadores, concorrentes e/ou agentes microbianos. Esta nova abordagem fornece importantes subsídios para o desenvolvimento de novos agentes de controle biológico ou produtos de base microbiana, com o potencial de reduzir a carga parasitária transportada pelo vetor esquistossomose de caramujos e controlar sua transmissão de forma sustentável.

Phenotypic and biochemical characterisation and pathogenicity assessment on Galleria mellonellaL. (Lepidoptera: Pyralidae) of symbionts of the entomopathogenic nematode Heterorhabditis amazonensis Andalo et al., 2006 / Caracterização fenotípica, bioquímica e avaliação da patogenicidade à Galleria mellonella L. (Lepidoptera: Pyralidae) de bactérias simbiontes do nematoide entomopatogênico Heterorhabditis amazonensisAndalo et al., 2006

The objective of this study was to describe phenotypically and biochemically the symbiotic bacteria associated with three populations of Heterorhabditis amazonensis Andalo et al., 2006 (isolates: UEL-n 01, UEL-n 07, and UEL-n 08) and evaluate their pathogenicity on Galleria mellonella L. (Lepidoptera: Pyralidae) larvae. Bacteria were isolated by maceration of infective juveniles (IJs) and grown in culture medium (NBTA and MacConkey). The characterization of the bacteria was evaluated by employing motility test and biochemical tests like Gram staining, lipase activity, protease, and lecithinase. The production of antibiotics and bioluminescence was also evaluated. The pathogenicity was evaluated on the last instar larvae of G. mellonella at a concentration of 104 cells/mL. The bacteria from the three entomopathogenic nematodes isolates were positive for all biochemical tests except for lecithinase, and have presented bioluminescence when subjected to ultraviolet light, indicating that they belong to the genus Photorhabdus sp. Both were pathogenic to G. mellonella larvae causing 93.3 to 100.0% mortality.(AU)

O objetivo desse trabalho foi descrever fenotípica e bioquimicamente as bactérias simbiontes associadas a três isolados do nematoide Heterorhabditis amazonensis Andalo et al., 2006 (isolates: UEL-n 01, UEL-n 07, and UEL-n 08) e avaliar sua patogenicidade sobre lagartas de Galleria mellonella L. (Lepidoptera: Pyralidae). As bactérias foram isoladas por meio de macerado de juvenis infectantes (JIs) e cultivadas em meio de cultura (NBTA e MacConkey). A caracterização das bactérias foi realizada por meio de testes de motilidade e bioquímicos de coloração de Gram, atividade de lipase, protease e lecitinase. Avaliou-se também a produção de antibióticos e de bioluminescência. A patogenicidade foi avaliada em lagartas de G. mellonella na concentração de 104 células/mL. Observou-se que as bactérias dos três isolados de nematoides entomopatogênicos foram positivas para todos os testes bioquímicos, com exceção da lecitinase, e apresentaram bioluminescência quando submetidas a luz ultravioleta, indicando que estas cepas pertencem ao gênero Photorhabdus sp. Ambas foram patogênicas às lagartas de G. mellonellacausando mortalidade entre 93,3 e 100,0%.(AU)

Biological control of Biomphalaria, the intermediate host of Schistosoma spp, a systematic review / Controle biológico de Biomphalaria, hospedeiro intermediário do Schistosoma spp, uma revisão sistemática

Schistosomiasis is an important vector-borne disease transmitted by an intermediate host: a freshwater mollusk. Control of these snail vectors is one of the strategies of the World Health Organization against the disease. The present study was based on a systematic review of published scientific papers concerning the biological control of snails (genus Biomphalaria), and identified the ongoing challenges and propose future perspectives. The review methodology was based on the PRISMA statement, the international databases Web of Science and Scopus for the period 1945-2021. In total, 47 papers were analyzed, published by authors from 14 different countries, the majority being from: France, Brazil, the United States, and Egypt. The most widely used strategy for biological control was predation by fish (12 studies). Fourteen papers were published in the most prolific decade 2010-2019; during which there was also a greater diversity of biological control agents in studies. In this context, we believed that one of the principal challenges of this approach is the successful simultaneous use of multiple types of biological control agent: predators, competitors, and/or microbial agents. This new approach may provide important insights for the development of new biological control agents or microbial-based products, with the potential to reduce the parasite load carried by schistosomiasis snail vector and control its transmission in a sustainable way.

A esquistossomose é uma importante doença transmitida por vetor, um hospedeiro intermediário: um molusco de água doce. O controle desses caramujos vetores é uma das estratégias da Organização Mundial da Saúde para controle da doença. O presente estudo foi baseado em uma revisão sistemática de artigos científicos publicados sobre o controle biológico de caramujos (gênero Biomphalaria), e teve como objetivo identificar os desafios atuais e propor perspectivas futuras. A metodologia de revisão foi baseada na declaração PRISMA, nas bases de dados internacionais, Web of Science e Scopus, entre 1945-2021. No total, foram analisados 47 artigos, publicados por autores de 14 países diferentes, sendo a maioria: França, Brasil, Estados Unidos e Egito. A estratégia mais utilizada para controle biológico foi a predação por peixes (12 estudos). Quatorze artigos foram publicados na década mais produtiva 2010-2019, durante a qual também houve uma maior diversidade de agentes de controle biológico em estudos. Neste contexto, acreditamos que um dos principais desafios desta abordagem é a utilização simultânea bem-sucedida de múltiplos tipos de agentes de controle biológico: predadores, concorrentes e/ou agentes microbianos. Esta nova abordagem fornece importantes subsídios para o desenvolvimento de novos agentes de controle biológico ou produtos de base microbiana, com o potencial de reduzir a carga parasitária transportada pelo vetor esquistossomose de caramujos e controlar sua transmissão de forma sustentável.

Susceptibility of embryos of Biomphalaria tenagophila (Mollusca: Gastropoda) to infection by Pochonia chlamydosporia (Ascomycota: Sordariomycetes).

Schistosoma mansoni is a heteroxenous parasite, meaning that during its life cycle needs the participation of obligatory intermediate and definitive hosts. The larval development occurs in aquatic molluscs belonging to the Biomphalaria genus, leading to the formation of cercariae, which emerge to infect the final vertebrate host. For this reason, studies for control of the diseases caused by digenetic trematodes often focus on combating the snail hosts. Thus, the objective of this study was to evaluate the susceptibility of Biomphalaria tenagophila embryos to the fungus Pochonia chlamydosporia (isolate Pc-10). The entire experiment was conducted in duplicate, with five replicates for each repetition (five egg masses/replicate), utilizing a total of 100 egg masses, with 20-30 eggs/egg mass. At the end of 15 days, the egg masses were evaluated under a stereomicroscope to analyze the hatching of B. tenagophila embryos in both experimental groups. After days of interaction, the exposure to the fungal hyphae bodies significantly impaired the viability of the B. tenagophila eggs, inhibiting the embryogenesis process by 83.7% in relation to the control group. Transmission and scanning electron microscopic images revealed relevant structural alterations in the egg masses exposed to the hyphae action of the fungus, interfering in the development and hatching of the young snails under analysis. These results indicate the susceptibility of B. tenagophila embryos to the fungus P. chlamydosporia (isolate Pc-10) and suggest the potential of Pc-10 to be used in the control of intermediate host, for its ovicidal capacity and for being an ecologically viable option, but in vivo experiments become necessary.

Heterorhabditis bacteriophora (Rhabditida: Heterorhabditidae), isolate HP88, induces reproductive and physiological alterations in Biomphalaria glabrata (Gastropoda: Planorbidae): an alternative for biological control of schistosomiasis.

Heterorhabditis bacteriophora is an entomopathogenic nematode (EPN) that is mutually associated with Photorhabdus luminescens, utilized globally for biological control of numerous organisms. Freshwater snails of the species Biomphalaria glabrata have been incriminated as the main intermediate hosts of Schistosoma mansoni in Brazil, but virtually nothing is known about the susceptibility of these gastropod to EPNs. Information in this respect is relevant for control of these intermediate hosts, and thus of the helminthiases they transmit. This paper for the first time reports the susceptibility of B. glabrata to infective juveniles of H. bacteriophora (isolate HP88) under laboratory conditions. For that purpose, six groups were formed: three Control groups (not exposed) and three Treated groups, in which the snails were exposed to 300 juveniles infecting the nematode over three weeks. The entire experiment was conducted in triplicate, using a total of 270 snails. Significant physiological alterations in B. glabrata were observed in response to the infection by H. bacteriophora HP88, characterized by decreased levels of hemolymphatic glucose as well as reduced contents of glycogen stored in the host's digestive gland. In parallel, the hemolymphatic activity of lactate dehydrogenase increased in the infected snails, indicating that the infection induces breakdown of carbohydrate homeostasis in B. glabrata. Additionally, all the reproductive parameters analyzed were reduced as a consequence of the infection. The results indicate the occurrence of the phenomenon of parasitic castration in the B. glabrata/H. bacteriophora HP88 interface, probably due to the depletion of galactogen in the parasitized organism. Although the infection did not cause lethality in the population of infected snails, H. bacteriophora HP88 compromised the reproductive performance of B. glabrata, suggesting its applicability in programs for biological control of this planorbid.

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.

Maintenance of sterility in SMS packaging in dental environments: concern with biosafety

Healthcare services must be guided by biosafety practices and microbial control. This control is highly influenced by humidity, which directly impacts the maintenance of sterility of the materials used in the appointments. High concentration of moisture, in the form of aerosol, splashes and spills, is caused during dental care. During the COVID-19 times the contamination by aerosol and droplets worries greatly. Considering that it could cause harm to the sterility of an autoclaved material, especially in dental environments, the objective was to evaluate the behavior of SMS sterilization packages (Spunbonded / Meltblown / Spunbonded) against microbial penetration in an aqueous vehicle. SMS of three brands were challenged, equally divided into two groups: virgin and processed (subjected to a single autoclaving cycle). Each specimen was aseptically deposited on Macconkey agar. Subsequently, 5 µL of Escherichia coliATCC 25922 saline solution [108CFU mL-1] was deposited in center of the SMS specimen and the dish incubated at36°C/ 48h. Reading was performed by the presence or absence of bacterial growth typical of the species under the SMS, observed on the back of Petri dish. The lowest penetration rate observed was 60% for one of the brands in the virgin condition, and 75% for two brands in the processed condition. Statistical analysis showed an association between bacterial penetration and the evaluated group, this association being valid only in the virgin condition. The different SMS behave similarly in terms of resistance to bacterial penetration after being processed. The data show that moisture can assist in bacterial transport through sterilized SMS. Therefore, SMS packages are not able to prevent bacterial penetration, and possibly other microorganisms, when in aqueous vehicles, offering a potential risk of breaking the aseptic chain. Thus, care must be taken in routines for handling and storage sterile packaging.

Ultrastructural and pathogenicity of Brevibacillus laterosporus against sinantropic muscoid dipterans.

Brevibacillus laterosporus has entomopathogenic potential against several orders of insects and its wide bioactivity is associated with a variety of strain-specific molecules. In order to avoid the use of synthetic insecticides, along with the need to control insect pests, microbial control has been widely used. Muscoid dipterans are known for their medical-veterinary and sanitary importance, and synanthropy. The enormous biotechnological potential of B. laterosporus has been demonstrated, but there are still few studies with muscoid dipterans. The aim of the study was to verify the mortality of B. laterosporus NRS590 on synanthropic flies and to characterize its different cell stages ultrastructurally. The flies were collected from garbage bins and the colonies were adapted to the laboratory conditions. Bioassays with neo larvae were carried out from the bacterial growth in the phases: vegetative (6 hr), sporangium (20 hr), and free spores (44 hr). An aliquot of each phase was collected for Transmission (TEM) and Scanning Electron Microscopy (SEM). The effectiveness of NRS590 was observed in the sporulation phase, where the corrected mortality was 83.3, 85.1, and 99% for Chrysomya megacephala, Chrysomya putoria, and Musca domestica, respectively. The parasporal body was observed in detail on the entire spore surface. Although our knowledge of this bacterium is growing, it remains to be determined the real virulence factors responsible for the wide entomopathogenic activity observed on muscoid dipterans. Therefore, this study can provide subsidies for the improvement of efficient and safe microbial control techniques for the environment and living beings.

A comparative study on the virulence of entomopathogenic fungi against Trogoderma granarium (Everts) (Coleoptera: Dermestidae) in stored grains rice / Um estudo comparativo da virulência de fungos entomopatogênicos contra Trogoderma granarium (Everts) (Coleoptera: Dermestidae) em grãos armazenados de arroz

Entomopathogenic fungi (EPF) now a possible safer microbial control measure that could be considered as a substitute for chemical control of insect pests. Three EPF viz., Metarihizium anisopliae, Isaria furnosoroseus and Beauveria bassiana were evaluated for their virulence against the grubs of Khapra beetle, Trogoderma granarium (Everts) under laboratory conditions. The isolates were applied by two methods viz., diet incorporation and an immersion method with 3rd instar 20 grubs of T. granarium for each. The virulence of EPF was determined using percent mortality. Significantly higher mortality was observed in M. anisopliae applied through immersion (98.33%) and diet incorporation (93.33%) methods followed by B. bassiana (90.83 and 85.83%, respectively). The mortality caused by I. furnosoroseus was statistically lower in immersion and diet incorporation methods i.e. 81.67 and 73.33%, respectively. Based on the immersion method, all EPF were studied for multiple conidial concentration i.e., 1×104, 1×105, 1×106, 1×107 and 1×108 under the same in-vitro conditions. All the isolates were pathogenic to grub of T. granarium at the highest conidial concentration. M. anisopliae was proved the most effective virulent resulting in 98.33% mortality of the pest with LT50 4.61 days at 1 × 108 conidial concentration followed by 90.83 and 81.67 percent mortality with 5.07 and 8.01 days LT50, in the application of B. bassiana and I. furnosoroseus, respectively. M. anisopliae showed higher efficacy and could be considered as promising EPF for the development of myco-insecticides against effective biocontrol of T. granarium.(AU)

Os fungos entomopatogênicos (FPE) são agora a possível medida de controle microbiano mais segura, que pode ser considerada um substituto para o controle químico de pragas de insetos. Três EPF viz., Metarihizium anisopliae, Isaria furnosoroseus e Beauveria bassiana foram avaliados quanto à sua virulência contra as larvas do besouro Khapra, Trogoderma granarium (Everts) em condições de laboratório. Os isolados foram aplicados por dois métodos, a saber: incorporação de dieta e um método de imersão com 20 larvas de T. granarium de 3º ínstar para cada um. A virulência do EPF foi determinada usando a mortalidade percentual. Mortalidade significativamente maior foi observada em M. anisopliae aplicado pelos métodos de imersão (98,33%) e incorporação de dieta (93,33%), seguido por B. bassiana (90,83% e 85,83%, respectivamente). A mortalidade causada por I. furnosoroseus foi estatisticamente menor nos métodos de imersão e incorporação de dieta, ou seja, 81,67% e 73,33%, respectivamente. Com base no método de imersão, todos os EPFs foram estudados para múltiplas concentrações de conídios, ou seja, 1 × 104, 1 × 105, 1 × 106, 1 × 107 e 1 × 108 nas mesmas condições in vitro. Todos os isolados foram patogênicos à larva de T. granarium na maior concentração de conídios. M. anisopliae provou ser o virulento mais eficaz, resultando em 98,33% de mortalidade da praga com LT50 4,61 dias na concentração de 1 × 108 conídios seguido por 90,83% e 81,67% de mortalidade com 5,07 e 8,01 dias LT50, na aplicação de B. bassiana e I. furnosoroseus, respectivamente. M. anisopliae apresentou maior eficácia e pode ser considerada como um PFE promissor para o desenvolvimento de micoinseticidas contra o biocontrole efetivo de T. granarium.(AU)

A comparative study on the virulence of entomopathogenic fungi against Trogoderma granarium (Everts) (Coleoptera: Dermestidae) in stored grains rice / Um estudo comparativo da virulência de fungos entomopatogênicos contra Trogoderma granarium (Everts) (Coleoptera: Dermestidae) em grãos armazenados de arroz

Entomopathogenic fungi (EPF) now a possible safer microbial control measure that could be considered as a substitute for chemical control of insect pests. Three EPF viz., Metarihizium anisopliae, Isaria furnosoroseus and Beauveria bassiana were evaluated for their virulence against the grubs of Khapra beetle, Trogoderma granarium (Everts) under laboratory conditions. The isolates were applied by two methods viz., diet incorporation and an immersion method with 3rd instar 20 grubs of T. granarium for each. The virulence of EPF was determined using percent mortality. Significantly higher mortality was observed in M. anisopliae applied through immersion (98.33%) and diet incorporation (93.33%) methods followed by B. bassiana (90.83 and 85.83%, respectively). The mortality caused by I. furnosoroseus was statistically lower in immersion and diet incorporation methods i.e. 81.67 and 73.33%, respectively. Based on the immersion method, all EPF were studied for multiple conidial concentration i.e., 1×104 , 1×105, 1×106, 1×107 and 1×108 under the same in-vitro conditions. All the isolates were pathogenic to grub of T. granarium at the highest conidial concentration. M. anisopliae was proved the most effective virulent resulting in 98.33% mortality of the pest with LT50 4.61 days at 1 × 108 conidial concentration followed by 90.83 and 81.67 percent mortality with 5.07 and 8.01 days LT50, in the application of B. bassiana and I. furnosoroseus, respectively. M. anisopliae showed higher efficacy and could be considered as promising EPF for the development of myco-insecticides against effective biocontrol of T. granarium.

Os fungos entomopatogênicos (FPE) são agora a possível medida de controle microbiano mais segura, que pode ser considerada um substituto para o controle químico de pragas de insetos. Três EPF viz., Metarihizium anisopliae, Isaria furnosoroseus e Beauveria bassiana foram avaliados quanto à sua virulência contra as larvas do besouro Khapra, Trogoderma granarium (Everts) em condições de laboratório. Os isolados foram aplicados por dois métodos, a saber: incorporação de dieta e um método de imersão com 20 larvas de T. granarium de 3º ínstar para cada um. A virulência do EPF foi determinada usando a mortalidade percentual. Mortalidade significativamente maior foi observada em M. anisopliae aplicado pelos métodos de imersão (98,33%) e incorporação de dieta (93,33%), seguido por B. bassiana (90,83% e 85,83%, respectivamente). A mortalidade causada por I. furnosoroseus foi estatisticamente menor nos métodos de imersão e incorporação de dieta, ou seja, 81,67% e 73,33%, respectivamente. Com base no método de imersão, todos os EPFs foram estudados para múltiplas concentrações de conídios, ou seja, 1 × 104, 1 × 105, 1 × 106, 1 × 107 e 1 × 108 nas mesmas condições in vitro. Todos os isolados foram patogênicos à larva de T. granarium na maior concentração de conídios. M. anisopliae provou ser o virulento mais eficaz, resultando em 98,33% de mortalidade da praga com LT50 4,61 dias na concentração de 1 × 108 conídios seguido por 90,83% e 81,67% de mortalidade com 5,07 e 8,01 dias LT50, na aplicação de B. bassiana e I. furnosoroseus, respectivamente. M. anisopliae apresentou maior eficácia e pode ser considerada como um PFE promissor para o desenvolvimento de micoinseticidas contra o biocontrole efetivo de T. granarium.

A comparative study on the virulence of entomopathogenic fungi against Trogoderma granarium (Everts) (Coleoptera: Dermestidae) in stored grains rice / Um estudo comparativo da virulência de fungos entomopatogênicos contra Trogoderma granarium (Everts) (Coleoptera: Dermestidae) em grãos armazenados de arroz

Entomopathogenic fungi (EPF) now a possible safer microbial control measure that could be considered as a substitute for chemical control of insect pests. Three EPF viz., Metarihizium anisopliae, Isaria furnosoroseus and Beauveria bassiana were evaluated for their virulence against the grubs of Khapra beetle, Trogoderma granarium (Everts) under laboratory conditions. The isolates were applied by two methods viz., diet incorporation and an immersion method with 3rd instar 20 grubs of T. granarium for each. The virulence of EPF was determined using percent mortality. Significantly higher mortality was observed in M. anisopliae applied through immersion (98.33%) and diet incorporation (93.33%) methods followed by B. bassiana (90.83 and 85.83%, respectively). The mortality caused by I. furnosoroseus was statistically lower in immersion and diet incorporation methods i.e. 81.67 and 73.33%, respectively. Based on the immersion method, all EPF were studied for multiple conidial concentration i.e., 1×104, 1×105, 1×106, 1×107 and 1×108 under the same in-vitro conditions. All the isolates were pathogenic to grub of T. granarium at the highest conidial concentration. M. anisopliae was proved the most effective virulent resulting in 98.33% mortality of the pest with LT50 4.61 days at 1 × 108 conidial concentration followed by 90.83 and 81.67 percent mortality with 5.07 and 8.01 days LT50, in the application of B. bassiana and I. furnosoroseus, respectively. M. anisopliae showed higher efficacy and could be considered as promising EPF for the development of myco-insecticides against effective biocontrol of T. granarium.

Os fungos entomopatogênicos (FPE) são agora a possível medida de controle microbiano mais segura, que pode ser considerada um substituto para o controle químico de pragas de insetos. Três EPF viz., Metarihizium anisopliae, Isaria furnosoroseus e Beauveria bassiana foram avaliados quanto à sua virulência contra as larvas do besouro Khapra, Trogoderma granarium (Everts) em condições de laboratório. Os isolados foram aplicados por dois métodos, a saber: incorporação de dieta e um método de imersão com 20 larvas de T. granarium de 3º ínstar para cada um. A virulência do EPF foi determinada usando a mortalidade percentual. Mortalidade significativamente maior foi observada em M. anisopliae aplicado pelos métodos de imersão (98,33%) e incorporação de dieta (93,33%), seguido por B. bassiana (90,83% e 85,83%, respectivamente). A mortalidade causada por I. furnosoroseus foi estatisticamente menor nos métodos de imersão e incorporação de dieta, ou seja, 81,67% e 73,33%, respectivamente. Com base no método de imersão, todos os EPFs foram estudados para múltiplas concentrações de conídios, ou seja, 1 × 104, 1 × 105, 1 × 106, 1 × 107 e 1 × 108 nas mesmas condições in vitro. Todos os isolados foram patogênicos à larva de T. granarium na maior concentração de conídios. M. anisopliae provou ser o virulento mais eficaz, resultando em 98,33% de mortalidade da praga com LT50 4,61 dias na concentração de 1 × 108 conídios seguido por 90,83% e 81,67% de mortalidade com 5,07 e 8,01 dias LT50, na aplicação de B. bassiana e I. furnosoroseus, respectivamente. M. anisopliae apresentou maior eficácia e pode ser considerada como um PFE promissor para o desenvolvimento de micoinseticidas contra o biocontrole efetivo de T. granarium.

A comparative study on the virulence of entomopathogenic fungi against Trogoderma granarium (Everts) (Coleoptera: Dermestidae) in stored grains rice

Abstract Entomopathogenic fungi (EPF) now a possible safer microbial control measure that could be considered as a substitute for chemical control of insect pests. Three EPF viz., Metarihizium anisopliae, Isaria furnosoroseus and Beauveria bassiana were evaluated for their virulence against the grubs of Khapra beetle, Trogoderma granarium (Everts) under laboratory conditions. The isolates were applied by two methods viz., diet incorporation and an immersion method with 3rd instar 20 grubs of T. granarium for each. The virulence of EPF was determined using percent mortality. Significantly higher mortality was observed in M. anisopliae applied through immersion (98.33%) and diet incorporation (93.33%) methods followed by B. bassiana (90.83 and 85.83%, respectively). The mortality caused by I. furnosoroseus was statistically lower in immersion and diet incorporation methods i.e. 81.67 and 73.33%, respectively. Based on the immersion method, all EPF were studied for multiple conidial concentration i.e., 1×104, 1×105, 1×106, 1×107 and 1×108 under the same in-vitro conditions. All the isolates were pathogenic to grub of T. granarium at the highest conidial concentration. M. anisopliae was proved the most effective virulent resulting in 98.33% mortality of the pest with LT50 4.61 days at 1 × 108 conidial concentration followed by 90.83 and 81.67 percent mortality with 5.07 and 8.01 days LT50, in the application of B. bassiana and I. furnosoroseus, respectively. M. anisopliae showed higher efficacy and could be considered as promising EPF for the development of myco-insecticides against effective biocontrol of T. granarium.

Resumo Os fungos entomopatogênicos (FPE) são agora a possível medida de controle microbiano mais segura, que pode ser considerada um substituto para o controle químico de pragas de insetos. Três EPF viz., Metarihizium anisopliae, Isaria furnosoroseus e Beauveria bassiana foram avaliados quanto à sua virulência contra as larvas do besouro Khapra, Trogoderma granarium (Everts) em condições de laboratório. Os isolados foram aplicados por dois métodos, a saber: incorporação de dieta e um método de imersão com 20 larvas de T. granarium de 3º ínstar para cada um. A virulência do EPF foi determinada usando a mortalidade percentual. Mortalidade significativamente maior foi observada em M. anisopliae aplicado pelos métodos de imersão (98,33%) e incorporação de dieta (93,33%), seguido por B. bassiana (90,83% e 85,83%, respectivamente). A mortalidade causada por I. furnosoroseus foi estatisticamente menor nos métodos de imersão e incorporação de dieta, ou seja, 81,67% e 73,33%, respectivamente. Com base no método de imersão, todos os EPFs foram estudados para múltiplas concentrações de conídios, ou seja, 1 × 104, 1 × 105, 1 × 106, 1 × 107 e 1 × 108 nas mesmas condições in vitro. Todos os isolados foram patogênicos à larva de T. granarium na maior concentração de conídios. M. anisopliae provou ser o virulento mais eficaz, resultando em 98,33% de mortalidade da praga com LT50 4,61 dias na concentração de 1 × 108 conídios seguido por 90,83% e 81,67% de mortalidade com 5,07 e 8,01 dias LT50, na aplicação de B. bassiana e I. furnosoroseus, respectivamente. M. anisopliae apresentou maior eficácia e pode ser considerada como um PFE promissor para o desenvolvimento de micoinseticidas contra o biocontrole efetivo de T. granarium.

Field Efficacy of Metarhizium rileyi Applications Against Spodoptera frugiperda (Lepidoptera: Noctuidae) in Maize.

The efficacy of the fungus Metarhizium rileyi (Farlow) Kepler, S. A. Rehner & Humber (Hypocreales: Clavicipitaceae) against the fall armyworm Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) and the impact on the community of epigeans predators as well as the natural occurrence of native fungal strains were evaluated after sequential sprays in maize. In the first season, the infestation was lower than 0.2 larvae per plant throughout the three fungal sprays and no significant difference was observed in the number of infected larvae among treatments. In the second season, insect population reached an average of 0.8 larvae per plant between the second and third sprays and the number of larvae succumbed by M. rileyi was higher in fungal-treated plots in the following weeks. Molecular characterization of the collected isolates on mummified larvae revealed the introduction of a new isolate did not eliminate the native strain on the fungal-treated plots. In both seasons, the diversity and abundance of predator species in the plots were not affected by any of the treatments. We showed that applications of selected strains of M. rileyi early in the season have no significant impact on the occurrence of natural enemies in maize.

Sub-lethal effects of a Bt-based bioinsecticide on the biological conditioning of Anticarsia gemmatalis.

Bioinsecticides based on Bacillus thuringiensis (Bt) Berliner, 1915 are widely used to control lepidopteran in several crops. However, surviving insects exposed to the sub-lethal concentration of Bt-based bioinsecticides can suffer a multitude of effects on the biological conditioning known as hormesis. Here, we aimed to provide a clearer understanding of the biological conditioning of Anticarsia gemmatalis (Hübner, 1818), exposed to different concentrations of a Bt-based bioinsecticide, by assessing life table parameters over three generations. We defined five sub-lethal concentrations (LC5, LC10, LC15, LC20, and LC25) from the response curve estimate of A. gemmatalis. Deionized water was used as a control. We assessed the parameters of eggs-viability and the duration of the stages, incubation, larval, pre-pupal, pupal, adult, pre-oviposition and total biological cycle. Data were used to construct the fertility life table using the two-sex program. The survival curves showed greater variation in the proportion of individuals at each development stage using the LC25. The sub-lethal concentrations did not influence the incubation-eggs period, pre-pupal and pupal. However, the larval and adult stages using LC25 and LC10 were the most affected. Changes in sex ratio were observed using LC20 and LC5. The toxic effect of Bt-based bioinsecticide interfered mainly in the parameters of fertility, sex ratio, net reproduction rate (R0), and gross reproduction rate (GRR).

Encapsulation Strategies for Bacillus thuringiensis: From Now to the Future.

Bacillus thuringiensis (Bt) has been recognized for its high potential in the control of various agricultural pests. Developments in micro/nanotechnology have opened new perspectives for the production of more efficient formulations that can overcome some obstacles associated with its use in the field, such as formulation instability and loss of activity as a result of the degradation of pesticidal protein by its exposure to ultraviolet radiation, among other problems. This review describes current studies and recent discoveries related to Bt and processes for the encapsulation of Bt derivatives, such as Cry pesticidal proteins. Different techniques are described, such as extrusion, emulsion, spray drying, spray cooling, fluidized bed, lyophilization, coacervation, and electrospraying to obtain micro- and nanoparticulate systems. It is noteworthy that products based on microorganisms present less risk to the environment and non-target organisms. However, systematic risk assessment studies of these new Bt biopesticides are necessary, considering issues, such as interactions with other organisms, the formation of toxic secondary metabolites, or the interspecific transfer of genetic material. Given the great potential of these new formulations, a critical assessment is provided for their future use, considering the technological challenges that must be overcome to achieve their large-scale production for efficient agricultural use.

Effects of Trichoderma strigosellum in Eucalyptus urophylla Development and Leaf-Cutting Ant Behavior.

Fungal endophytes can protect plants against herbivory and be used to control leaf-cutting ants. In this study, we aimed to evaluate the potential of endophytic colonization of Eucalyptus urophylla by three filamentous fungal species and their influence on the plant development and foraging behavior of Atta sexdens. The study design was completely randomized and comprised a factorial scheme of 4 × 3, three antagonistic fungal species (Escovopsis sp., Metarhizium anisopliae, and Trichoderma strigosellum) of the leaf-cutting ant, and one control and three inoculation methods (conidial suspension via foliar spray [FS] and soil drench [SD] inoculation, and seedlings inoculated with mycelium [SWM]). The SWM method allowed T. strigosellum to colonize all plant organs, and these plants exhibited higher height, leaf number, shoot dry mass, and total dry mass than the ones subjected to the other inoculation methods. The SWM method increased the plant height than the control plants and those inoculated with Escovopsis sp. and M. anisopliae. Trichoderma strigosellum, previously isolated from soil, colonized E. urophylla plants and positively influenced their development, as demonstrated by the SWM method. Trichoderma strigosellum promoted the increase in E. urophylla height compared with when the FS and SD methods were used (by 19.62% and 18.52%, respectively). Our results reveal that A. sexdens workers preferentially began cutting the leaves from plants not previously colonized by T. strigosellum. This behavior can be explained by modifications in the phenotypic traits of the eucalyptus leaves.

Characterization of antimicrobial effect of organotin-based catalysts on diesel-biodiesel deteriogenic microorganisms.

Organotin compounds are applied in several industrial reactions and can present antifungal and antibacterial activities. Incorrect handling and storage practices of biodiesel and diesel-biodiesel blends can lead to microbial development, impacting its final quality. Concerning this problem, this work investigated the antimicrobial action of two organotin catalysts used in biodiesel production with four isolated microroorganisms (Bacillus pumilus, Pseudomonas aeruginosa, Pseudallescheria boydii, and Aureobasidium pullulans) and a pool of microorganisms (ASTM E1259 standard practice). Samples of soybean biodiesel with different concentrations of dibutyl tin dilaurate (catalyst 1) and di-n-butyl-oxo-stannane (catalyst 2) were prepared and added of mineral medium. The pool of microorganisms was inoculated and incubated at 30 °C and final biomass was weighted after 14 days. Thereafter, soybean biodiesel with catalyst 2 was used. Fungal biomass was weighted, and plate count was used to assess bacterial growth. Results show that catalysts 1 and 2 presented no inhibitory activity on the pool of microorganisms evaluated. A slight inhibitory activity was observed for B. pumilus and A. pullulans growth, but not for P. boydii, P. aeruginosa, or the pool of microorganisms. All experiment exhibited acidification higher than sterile control. Infrared analysis show less microbiological degradation products in the tin-protected fuel with ASTM inoculum. These results suggest that these tin-based catalysts show no toxic effect on native microbial population and a slight effect on some isolated microbial population in laboratory scale and for the first time shows that these organotin compounds can be employed safely as biodiesel catalyst. Graphical abstract.