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This study aimed to isolate and characterize a native strain of Beauveria bassiana, coded as Bv065, showcasing its potential as a biological control agent targeting the palm weevil Dynamis borassi. Originating from a naturally infected D. borassi specimen collected in southwestern Colombia, the fungus underwent molecular identification and was identified as B. bassiana, exhibiting high sequence similarity with known reference strains. The physiological characterization revealed that Bv065 thrived within a temperature range of 25 to 30 °C and a pH range of 6 to 9. Moreover, the key carbon sources that allow optimal growth of the strain were identified through metabolic profiling, including sucrose, D-mannose, and γ-amino-butyric acid. These findings offer strategic insights for scalability and formulation methodologies. Additionally, enzymatic analyses unveiled robust protease activity within Bv065, crucial for catalysing insect cuticle degradation and facilitating host penetration, thus accentuating its entomopathogenic potential. Subsequent evaluations exposed Bv065's pathogenicity against D. borassi, causing significant mortality within nine days of exposure, albeit exhibiting limited effectiveness against Rhynchophorus palmarum. This study underscores the importance of understanding optimal growth conditions and metabolic preferences of B. bassiana strains for developing effective biopesticides. The findings suggest Bv065 as a promising candidate for integrated pest management strategies in neotropical regions, particularly for controlling palm weevil infestations in coconut and peach palm cultivation. Future research avenues include refining mass production methodologies, formulating novel delivery systems, and conducting comprehensive field efficacy trials to unlock the full potential of Bv065 in fostering sustainable pest management practices. Overall, this study contributes to the growing body of knowledge on entomopathogenic fungi and their pivotal role in biological control, offering nuanced perspectives on eco-friendly alternatives to conventional insecticidal interventions., (© 2024. The Author(s).)

Bacillus thuringiensis (Bt) and entomopathogenic fungi (EPF) are in use for management of insect pests. Continuous use of Bt can lead to problem of resistance development in insect pests. Hence use of combination formulations (CF) of microbials with diverse modes of action has been attempted to slow down the process of resistance development. Suspension concentrate (SC) formulations of a local strain of Bt var. kurstaki DOR Bt-127 were developed singly and in combination with conidia of the EPF Nomuraea rileyi (Nr) and Beauveria bassiana (Bb). Electron microscopy of Bt + Bb CF treated larvae of Helicoverpa armigera revealed simultaneous infection by both microbials indicating their compatibility. Endotoxin contents in Bt-SC, Bt + Bb and Bt + Nr CFs were 5.0, 4.7 and 4.7%, respectively. These formulations were effective against larvae of Spodoptera litura, H. armigera and Achaea janata coupled with a lowering of the effective requirement of Bt and EPF. In multi-location field trials, Bt-SC and Bt + Nr CF were highly effective against S. litura and A. janata on castor as well as H. armigera and Thysanoplusia orichalcea on sunflower. However, Bt + Bb CF was highly effective only on sunflower against H. armigera and T. orichalcea. All formulations had 24 months shelf-life at room temperature. DOR Bt-127 based SC formulations developed singly and in combination with Nr and Bb were effective against major lepidopteran pests of castor and sunflower and did not lose viability under storage at room temperature. The CFs of Bt with EPF could prove promising for mitigating resistance development to Bt.

Beauveria bassiana (Bals.) Vuillemin (B. bassiana) is an entomopathogenic fungus that establishes endophytic symbiosis with plants. In the present study, the effects of B. bassiana strains colonization in growing Zea mays L. (Z. mays), crop production, and drought tolerance were evaluated. Z. mays seeds were inoculated with B. bassiana strains (GHA, PTG4, and PTG6), using 1 × 10 6 blastospores/mL and methyl cellulose (MC) or cornstarch (CS) as adherents. Colonization was determined by B. bassiana recovery from plant tissues plated on PDA medium. Plant height, fresh and dry weight, and flowering time were analyzed to assess plant performance. Drought tolerance was evaluated by stopping watering for 10 days, watering again, and determining vigor recovery after 24 h. Results showed 100% endophytic roots colonization, regardless of adherent type or strain tested. Colonization was variable in shoots and leaves, but GHA strain achieved the highest inoculation rates, including 88% in stems and 50% in leaves, which did not depend on adherent type used; for PTG4 strain, adherent type had an important effect (MC = 100% stems and leaves; CS = 63% stems and 25% leaves). For PTG6 strain, the best adherent type was CS (71% stems and 75% leaves), whereas MC showed variable inoculation percentage (25% stems and 75% leaves). Interestingly, only MCPTG4 treatment showed consistent positive effects on germination percentage (day 5 = 46 ± 2%; day 14 = 87 ± 7%) compared with controls (CC = 63 ± 4%, CMC = 50 ± 3%, CCS = 47 ± 0%). In addition, the other treatments showed low germination percentages at day 5 (7 ± 7% to 46 ± 2%), which recovered at day 14 (53 ± 0% to 73 ± 8%), except for MCPTG6 treatment with 23 ± 10% germination. About plant performance, not significant effects on plant height and fresh/dry weight in all the treatments were observed. However, B. bassiana-treated plants, using either GHA, PTG4 or PTG6 strains, and MC as adherent, showed tolerance to drought and flowered one to two weeks earlier, providing evidence supporting further applications of these seed treatments in agricultural systems, for abiotic stress sustainable management practices.

Beauveria bassiana plays a prominent role in biocontrol of houseflies, Musca domestica (L.). Thus, a deeper insight into immune response of M. domestica during B. bassiana infection was warranted to assist the production of more efficient mycoinsecticides. The present study investigates changes in protein profile of M. domestica hemolymph serum post B. bassiana infection using two-dimensional difference gel electrophoresis (2D-DIGE) followed by identification of selected proteins by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). The non-infected or control group of flies showed an expression of 54 proteins, while M. domestica infected with B. bassiana expressed a total of 68 hemolymph serum proteins. Thirty three proteins were expressed in both groups of houseflies, whereas 35 proteins were exclusively expressed in infected flies and 21 proteins were exclusively expressed in control flies. Among the 33 proteins which were expressed in both groups of houseflies, 17 proteins showed downregulation, while16 proteins were upregulated in the infected flies compared to the non-infected ones. The results from this study are expected to facilitate better understanding of insect's immune response mechanism.

The fungal entomopathogen Beauveria bassiana is a promising biocontrol agent for many pests. Some B. bassiana strains have been found effective against jute pests. To monitor the survival of field released B. bassiana a rapid and efficient detection technique is essential. Conventional methods such as plating method or direct culture method which are based on cultivation on selective media followed by microscopy are time consuming and not so sensitive. PCR based methods are rapid, sensitive and reliable. A single primer PCR may fail to amplify some of the strains. However, multiplex PCR increases the possibility of detection as it uses multiple primers. Therefore, in the present investigation a multiplex PCR protocol was developed by multiplexing three primers SCA 14, SCA 15 and SCB 9 to detect field released B. bassiana strains from soil as well as foliage of jute field. Using our multiplex PCR protocol all the five B. bassiana strains could be detected from soil and three strains viz., ITCC 6063, ITCC 4563 and ITCC 4796 could be detected even from the crop foliage after 45 days of spray.

Apoptosis and phagocytosis are crucial processes required for developmental morphogenesis, pathogen deterrence and immunomodulation in metazoans. We present data showing that amebocytes of the chelicerate, Limulus polyphemus, undergo phagocytosis-induced cell death after ingesting spores of the fungus, Beauveria bassiana, in vitro. The observed biochemical and morphological modifications associated with dying amebocytes are congruent with the hallmarks of apoptosis, including: extracellularisation of phosphatidylserine, intranucleosomal DNA fragmentation and an increase in caspase 3/7-like activities. Previous studies have demonstrated that phosphatidylserine is a putative endogenous activator of hemocyanin-derived phenoloxidase, inducing conformational changes that permit phenolic substrate access to the active site. Here, we observed extracellular hemocyanin-derived phenoloxidase activity levels increase in the presence of apoptotic amebocytes. Enzyme activity induced by phosphatidylserine or apoptotic amebocytes was reduced completely upon incubation with the phosphatidylserine binding protein, annexin V. We propose that phosphatidylserine redistributed to the outer plasma membrane of amebocytes undergoing phagocytosis-induced apoptosis could interact with hemocyanin, thus facilitating its conversion into a phenoloxidase-like enzyme, during immune challenge.

Isolates of the fungus Beauveria bassiana have different levels of virulence and repellency against the termite Macrotermes michaelseni. In the present study, we compared the volatile profiles and gene sequences of two isolates of the fungus with different levels of virulence and repellence to the termite. Gas chromatography-mass spectrometric analyses showed quantitative and qualitative differences in the composition of volatiles of the two isolates. The repellencies of synthetic blends of 10 prominent components that mimicked the volatiles of each of the two isolates were significantly different. Subtractive bioassays showed that the repellency of each isolate was due to synergistic effects of a few constituents. As previously reported for isolates of Metarhizium anisopliae, some differences also were found in the nucleotide sequences of the two isolates of B. bassiana, suggesting a genetic basis for the observed intra-specific differences in their repellency and virulence against the termite.

The effect of three levels of oxygen (normal atmosphere (21% O(2)), low oxygen (16% O(2)) and enriched oxygen (26% O(2))) on the production and germination of conidia by Beauveria bassiana was evaluated using rice as a substrate. The maximum yield of conidia was achieved under hypoxia (16% O(2)) after 8 days of culture (1.51 × 10(9) conidia per gram of initial dry substrate), representing an increase of 32% compared to the normal atmosphere. However, germination was reduced by at least 27% due to atmospheric modifications. Comparison of antioxidant enzyme activity (superoxide dismutases and catalases) with the oxidation profiles of biomolecules (proteins and lipids) showed that a decrease in catalase activity in the final days of culture coincided with an increase in the amount of oxidized lipids, showing that oxidative stress was a consequence of pulses of different concentrations of O(2). This is the first study describing oxidative stress induction by atmospheric modification, with practical implications for conidia production.

Trichogramma pretiosum Riley and the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae are efficient biological control agents and are thought to be used jointly. In here, we investigated if these entomopathogens could have any side-effects on T. pretiosum. Therefore, 1 x 8 cards containing sterilized eggs of Anagasta kuehniella (Zeller) that were sprayed with 0.2 ml of B. bassiana or M. anisopliae (1.0 x 10(9) conidia/ml) were offered to a T. pretiosum female for 24h (30 cards/fungus = 30 replicates). Afterwards, females were isolated in glass tubes. The control group was sprayed with sterile distillated water + Tween 80 (0.01%). In addition, 60 cards with sterilized eggs of A. kuehniella were submitted to parasitism by females of T. pretiosum for 24h. Of these cards, 30 were sprayed with B. bassiana or M. anisopliae and 30 with distillated water + Tween 80 (0.01%), and observed daily until parasitoid emergence. Metarhizium anisopliae decreased parasitoid emergence and caused confirmed mortality. Therefore, field and semi-field experiments should be conducted for a final assessment of the side-effects of these entomopathogens on Trichogramma as a ways to develop a control strategy in which both can be used.

To explore possible role of intracellular trehalose accumulation in fungal tolerance to summer-like thermal stress, 3-day colonies of Beauveria bassiana grown on a glucose-free medium at 25 degrees C were separately exposed to 35, 37.5 and 40 degrees C for 1-18 h, respectively. Trehalose accumulation in stressed mycelia increased from initial 4.2 to 88.3, 74.7 and 65.5 mg g(-1) biomass after 6-h stress at 35, 37.5 and 40 degrees C, respectively, while intracellular mannitol level generally declined with higher temperatures and longer stress time. The stress-enhanced trehalose level was significantly correlated to decreased trehalase activity (r(2) = 0.73) and mannitol content (r(2) = 0.38), which was inversely correlated to the activity of mannitol dehydrogenase (r(2) = 0.41) or mannitol 1-phosphate dehydrogenase (r(2) = 0.30) under the stresses. All stressed cultures were successfully recovered at 25 degrees C but their vigor depended on stressful temperature, time length and the interaction of both (r (2) = 0.98). The highest level of 6-h trehalose accumulation at 35 degrees C was found enhancing the tolerance of the stressed cultures to the greater stress of 48 degrees C. The results suggest that the trehalose accumulation result partially from metabolized mannitol and contribute to the fungal thermotolerance. Trehalase also contributed to the thermotolerance by hydrolyzing accumulated trehalose under the conditions of thermal stress and recovery.

Beetles share with other eukaryotes an innate immune system that mediates endogenous defense against pathogens. In addition, larvae of some taxa produce fluid exocrine secretions that contain antimicrobial compounds. In this paper, we provide evidence that larvae of the brassy willow leaf beetle Phratora vitellinae constitutively release volatile glandular secretions that combat pathogens in their microenvironment. We identified salicylaldehyde as the major component of their enveloping perfume cloud, which is emitted by furrow-shaped openings of larval glandular reservoirs and which inhibits in vitro the growth of the bacterial entomopathogen Bacillus thuringiensis. The suggested role of salicylaldehyde as a fumigant in exogenous antimicrobial defense was confirmed in vivo by its removal from glandular reservoirs. This resulted in an enhanced susceptibility of the larvae to infection with the fungal entomopathogens Beauveria bassiana and Metarhizium anisopliae. Consequently, we established the hypothesis that antimicrobial defense in beetles can be expanded beyond innate immunity to include external disinfection of their microenvironment, and we report for the first time the contribution of fumigants to antimicrobial defense in animals.

This study was carried out to evaluate entomopathogenic fungi isolates as microbial control agents ofthe lesser mealworm, Alphitobius diaperinus (Panzer). Larvae and adults were inoculated with conidial suspension of 99 isolates of Beauveria bassiana (Bals.) Vuill. and Metarhizium anisopliae (Metsch.) Sorok. (10(5) to 10(9) conidia/ml). Vegetative growth on culture media and sporulation on culture media, cooked rice and lesser mealworm cadavers were also evaluated. Isolates of B. bassiana were more effective than the M. anisopliae isolates and larvae were more susceptible than adults. The isolates UNIOESTE 04 and UNIOESTE 02 were originally obtained from adults of the lesser mealworm and were the most efficient isolates. In addition, UNIOESTE 04 showed high level of sporulation on different culture media and is considered a potential useful tool to lesser mealworm control.

Chemical insecticides are broadly applied to control diamondback moth, Plutella xylostella (L.). Diamondback moth is a major pest of cruciferous worldwide, and resistance of this pest to insecticide has been often reported. Thus, this research investigated the interactions among the fungi Metarhizium anisopliae (Metsch.) Sorok., Beauveria bassiana (Bals.) Vuill., and the larval-pupal parasitoid Oomyzus sokolowskii (Kurdjumov) before and after application of the fungi on DBM larvae offered to the parasitoid. The experiment was carried out at 26+/-l degreeC, 75+/- 5% RH and 12h photophase using a completely randomized design, with eight treatments with six replications each. The isolates E9 of M. anisopliae and ESALQ 447 of B. bassiana, were used at the concentration of 10(7) conidia ml(-1). The results showed that M. anisopliae and B. bassiana reduced the parasitism of P. xylostella by O. sokolowskii. Additive effects were found on the mortality of P. xylostella with the different combinations among the fungi and parasitoid, except for the treatment B. bassiana inoculated 24h before exposition of the larvae to O. sokolowskii. The isolates were more efficacious when applied after exposition of the larvae to the parasitoid. The efficiency of O. sokolowskii was negatively influenced by the presence of the fungi, mainly when the fungi were applied 24h before diamondback's larvae were exposed to the parasitoid. The association of the fungi with the parasitoid presents potential to be tested in field. The use of these natural enemies in the integrated management of P. xylostella may economically improve the cabbage productive system, especially for organic farming.