From lab to field: biological control of the Japanese beetle with entomopathogenic fungi.

The Japanese beetle, Popillia japonica, is an invasive scarab and listed as quarantine organism in many countries worldwide. Native to Japan, it has invaded North America, the Azores, and recently mainland Europe. Adults are gregarious and cause agricultural and horticultural losses by feeding on leaves, fruits, and flowers of a wide range of crops and ornamental plants. Larvae feed belowground and damage grassland. To date, no efficient and environmentally friendly control measure is available. Larval populations of other scarab species such as Phyllopertha horticola and Melolontha melolontha are controlled by applying spores of the entomopathogenic fungi Metarhizium brunneum and Beauveria brongniartii to larval habitats. Here, we tested this control strategy against Japanese beetle larvae in grasslands, as well as spore spray applications against adults in crops. Using both, large-scale field experiments and inoculation experiments in the laboratory, we assess the efficacy of registered fungal strains against Japanese beetle larvae and adults. Metarhizium brunneum BIPESCO 5 established and persisted in the soil of larval habitats and on the leaves of adult's host plants after application. However, neither larval nor adult population sizes were reduced at the study sites. Laboratory experiments showed that larvae are not susceptible to M. brunneum ART 212, M. brunneum BIPESCO 5, and B. brongniartii BIPESCO 2. In contrast, adults were highly susceptible to all three strains. When blastospores were directly injected into the hemolymph, both adults and larvae showed elevated mortality rates, which suggests that the cuticle plays an important role in determining the difference in susceptibility of the two life stages. In conclusion, we do not see potential in adapting the state-of-the-art control strategy against native scarabs to Japanese beetle larvae. However, adults are susceptible to the tested entomopathogenic fungi in laboratory settings and BIPESCO 5 conidiospores survived for more than three weeks in the field despite UV-radiation and elevated temperatures. Hence, control of adults using fungi of the genera Beauveria or Metarhizium is more promising than larval control. Further research on efficient application methods and more virulent and locally adapted fungal strains will help to increase efficacy of fungal treatments for the control of P. japonica.

The impact of Beauveria species bioinocula on the soil microbial community structure in organic strawberry plantations.

Introduction: The multifunctionality of microorganisms, including entomopathogenic fungi, represents a feature that could be exploited to support the development, marketing, and application of microbial-based products for plant protection. However, it is likely that this feature could affect the composition and dynamics of the resident soil microorganisms, possibly over a longer period. Therefore, the methodology utilized to evaluate such impact is critical for a reliable assessment. The present study was performed to evaluate the impact of strains of Beauveria brongniartii and Beauveria bassiana on soil bacterial and fungal communities using an approach based on the terminal restriction fragment polymorphism (T-RFLP) analysis. Materials and methods: Soil samples in the vicinity of the root system were collected during a 3-year period, before and after the bioinocula application, in two organic strawberry plantations. Specific primers were used for the amplification of the bacterial 16S rRNA gene and the fungal ITS region of the ribosome. Results and discussion: Data of the profile analysis from T-RFLP analysis were used to compare the operational taxonomic unit (OTU) occurrence and intensity in the inoculated soil with the uninoculated control. With regard to the impact on the bacterial community, both Beauveria species were not fully consistently affecting their composition across the seasons and fields tested. Nevertheless, some common patterns were pointed out in each field and, sometimes, also among them when considering the time elapsed from the bioinoculum application. The impact was even more inconsistent when analyzing the fungal community. It is thus concluded that the application of the bioinocula induced only a transient and limited effect on the soil microbial community, even though some changes in the structure dynamic and frequency of soil bacterial and fungal OTUs emerged.

Immune-Related Genes of Megalurothrips usitatus (Bagrall) Against Beauveria brongniartii and Akanthomyces attenuatus Identified Using RNA Sequencing.

Megalurothrips usitatus (Bagrall) is an important pest of legumes worldwide, causing great economic loss every year. Beauveria brongniartii and Akanthomyces attenuatus have shown considerable pathogenicity against M. usitatus in our previous studies. The medial lethal concentration (LC50) and the sublethal lethal concentration (LC25) of B. brongniartii isolate SB010 against M. usitatus were 8.38 × 105 and 1.73 × 105 conidia mL-1, respectively, whereas those of A. attenuatus isolate SCAUDCL-53 against M. usitatus were 4.37 × 105 and 2.97 × 104 conidia mL-1, respectively. This study reports the transcriptome-based explanation of the stress responses of M. usitatus following the application of B. brongniartii and A. attenuatus. The analysis of the transcriptomic data revealed the expression of 254, 207, 195, and 234 immunity-related unigenes by M. usitatus in response to B. brongniartii LC50 (SB1), B. brongniartii LC25 (SB2), A. attenuatus LC50 (V1), and A. attenuatus LC25 (V2), respectively. The biological function and metabolic pathway analyses showed that these unigenes were mainly related to pattern recognition receptors, information transduction factors, and reaction factors, such as scavenger receptor, cytochrome b5, cuticle protein, lysozyme, and serine protease.

Biocontrol of Melolontha spp. Grubs in Organic Strawberry Plantations by Entomopathogenic Fungi as Affected by Environmental and Metabolic Factors and the Interaction with Soil Microbial Biodiversity.

The efficacy of two strains of two Beauveria species (B. bassiana and B. brongniartii), individually or as co-inoculants, to control Melolontha sp. grubs was assessed in two organic strawberry plantations in relation to the environmental conditions, their abundance after soil inoculation, and their in vitro chitinolytic activity, thereby also verifying their impact on soil microbial communities. A reduction of the grubs' damage to strawberry plants was observed when compared to the untreated control in one plantation, irrespective of the strain used and whether they were applied as single or as co-inoculum. The metabolic pattern expressed by the two fungi in vitro was different: B. bassiana showed a higher metabolic versatility in the use of different carbon sources than B. brongniartii, whose profile was partly overlapped in the co-inoculum. Similar differences in the chitinolytic activity of each of the fungi and the co-inoculum were also pointed out. A higher abundance of B. bassiana in the soils receiving this species in comparison to those receiving B. brongniartii, together with its in vitro metabolic activity, could account for the observed diverse efficacy of pest damage control of the two species. However, environmental and climatic factors also affected the overall efficacy of the two bioinocula. According to the monitoring of the two species in soil, B. bassiana could be considered as a common native species in the studied locations in contrast to B. brongniartii, which seemed to be a non-endemic species. Nevertheless, the inoculation with both species or the co-inoculum did not consistently affect the soil microbial (fungi and bacteria) biodiversity, as expressed by the operational taxonomic unit (OTU) number and Shannon-Wiener diversity index based on terminal restriction fragment length polymorphism (TRFLP) data. A small transient increase of the share of the inoculated species to the total fungal community was noted by the analysis of genes copy numbers only for B. brongniartii at the end of the third growing season.

Biological Impact and Enzyme Activities of Spodoptera litura (Lepidoptera: Noctuidae) in Response to Synergistic Action of Matrine and Beauveria brongniartii.

Matrine, a naturally occurring heterocyclic compound, has been shown to enhance the pathogenicity of the entomopathogenic fungus Beauveria brongniartii against Spodoptera litura. In the current study, the biological impacts and synergism activities of these two agents on nutritional efficiency and antioxidant enzymes in S. litura were explored. Our results showed a high antifeedant activity of B. brongniartii and matrine on S. litura. The S. litura larvae were unable to pupate and emerge when treated with combinations of matrine and B. brongniartii. Following on, we measured the activities of five important antioxidant enzymes [superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), acetylcholinesterase (AChE), and glutathione-S-transferase (GST)] when treated with B. brongniartii SB010 (1 × 109 spores/ml), matrine (0.5 mg/ml), and B. brongniartii SB010 (1 × 109 spores/ml) + matrine (0.5 mg/ml). The results indicated the detoxification activity of the five enzymes in the fat body and hemolymph of S. litura when facing a combined B. brongniartii and matrine challenge. The activities of the enzymes were significantly lower than that of the control group 7 days post-treatment, indicating the inhibitory effect of the two xenobiotics. Matrine had better inhibition effects than B. brongniartii in a majority of the trials. The improved detoxification activity of the five enzymes may be the internal mechanism of synergism of matrine on B. brongniartii.

Screening of high-virulent entomopathogenic fungal strains to infect Xylotrechus rusticus larvae.

The gray tiger longicorn beetle, Xylotrechus rusticus Linnaeus (Coleoptera: Cerambycidae) is a stem-boring pest that can inhibit not only the transportation of nutrients in the trunk but also the tree growth, increasing the risk of tree breakage and causing economic losses. It is distributed in China, Iran, Turkey, Russia, Korea, Japan, and Southern Europe. This study aimed to investigate selected Beauveria strains that could be used as entomopathogenic fungi for the biological control of this pest. The high-virulence strains were screened among the selected strains by cumulative mortality, correct mortality, and lethal time to 50% mortality (LT50). These screened high-virulence strains were Bb01, CFCC83486, and CFCC81428. Bb01 exhibited 96.96% cumulative mortality, with an estimated LT50 of 3.28 days. CFCC83486 and CFCC81428 caused 89.29% and 75.74% cumulative mortality, with an estimated LT50 of 3.45 and 4.28 days, respectively. Pathogenicity at different concentrations and lethal concentration of 50% (LC50) of these high-virulence strains were investigated. The pathogenicity was found to be positively correlated with suspension concentration, and LC50 was negatively correlated with infection time. These suspensions of high-virulence strains at different concentrations were also investigated in the forest by brushing the suspensions on the poplar tree trunk infested with X. rusticus L. larvae. The most effective strain was found to be Bb01, whose cumulative mortality reached 76.33% at 1.32 × 108 conidia mL-1, followed by the strain CFCC83486, whose cumulative mortality reached 65.17% at 1.32 × 108 conidia mL-1. This study provides an important basis for using B. bassiana in the biological control of X. rusticus L.

Efecto de Beauveria bassiana y Beauveria brongniartii en el control de moscas (Musca domestica) en condiciones de laboratorio y en galpones avícolas / Effect of Beauveria bassiana and Beauveria brongniartii for the house fly (Musca domestica) control on laboratory and poultry sheds conditions

Se realizó un experimento en el estado Trujillo, Venezuela con el objetivo de evaluar en condiciones de laboratorio y en galpones avícolas la patogenicidad de Beauveria brongniartii (cepaLF-05) y Beauveria bassiana (LF-08) sobre Musca domestica. Se determinó que a la dosis de 1,2 x107 conidiosporas/mL para B. brongniartii y B. bassiana los tiempos letales (TL50 y TL95) fueron5 y 9 días y 5 y 6 días, respectivamente. En condiciones de campo las esporas de B. brongniartiiy B. bassiana fueron nebulizadas en el interior de galpones de cría de pollos con una densidad de 7 pollos/m2, a dosis de 9 x 107 conidias/mL en 15L de agua por cada 1200 m2; el porcentaje de reducción poblacional después de nebulizar una vez por semana, durante tres semanas, fue de 13,99 y 100% para B. brongniartii y -35, 91 y 100% para B. bassiana

An experiment was carried out in Trujillostate, Venezuela, in laboratory and poultry shedsconditions in order to study the Beauveria brongniartii(strain: LF-05) and Beauveria bassiana (strain: LF-08) pathogenicity in Musca domestica. With 1.2 x 107conidiospores/mL the lethal times (TL50 and TL95)were 5 and 9, and 5 and 6 days, respectively. In fieldconditions spores of B. brongniartii and B. bassianawere applied inside the poultry sheds (7 chickens/m2)with a dose of 9 x 107 conidia/mL in 15L of water foreach 1200 m2, the percentage of population reductionwere induced after vaporized once a week for threeweeks in 13.99 and 100% for B. brongniartii; and -35,91 and 100% for B. bassiana