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Metarhizium anisopliae (Metschnikoff) Sorokin and Beauveria bassiana (Balsamo) Vuillemin are entomopathogenic fungi commonly used in microbial control of arthropods. In this study, we evaluated the insecticidal potential of six isolates of B. bassiana (BIITAC10.3.3, BIITAC6.2.2, and BIITAC8.1.5) and M. anisopliae (MIITAC11.3.4, MIITAC6.2.2, and MIITAC6.4.2) from Cameroon, against the banana aphid Pentalonia nigronervosa Coquerel, the vector of the banana bunchy top virus (BBTV). Pathogenicity tests were initially conducted using B. bassiana and M. anisopliae isolates at a concentration of 3.2 × 106 conidia/ml on P. nigronervosa adults sourced from four agroecologies in Cameroon. Four isolates (BIITAC6.2.2, BIITAC10.3.3, BIITAC8.1.5, and MIITAC6.2.2) were highly pathogenic, causing greater than 75% aphid mortality in all populations. A significant decrease in aphid fecundity was observed with BIITAC6.2.2, MIITAC6.2.2, and BIITAC10.3.3. These three isolates were in a test of a series of four fungal concentrations (3.2 × 101, 3.2 × 102, 3.2 × 104, and 3.2 × 106 conidia/ml). produced LC50 of 1.31 × 101 and 3.12 × 10−2 for BIITAC10.3.3 and MIITAC6.2.2, respectively. MIITAC6.2.2 had the lowest LC90 (1.55 × 103). Our results strongly support the continued development of biopesticides based on one or more of the three fungal entomopathogens for the control of banana aphids as a component of an Integrated Pest Management (IPM) strategy for the reduction of the prevalence and transmission of BBTV under field conditions. [ABSTRACT FROM AUTHOR]

Both Metarhizium robertsii ARSEF 2575 (Mr2575) and Metarhizium anisopliae ARSEF 549 (Ma549) infect a range of insects whilst also interacting with plants; however, little is known about the traits that affect the competitive ability of different strains. We examined the interactions between Mr2575 and Ma549 in culture and during co-infection of plants (Arabidopsis thaliana) and insects. Mr2575 outcompetes Ma549 under nutrient-limiting conditions, including root exudates, giving it a priority advantage on Arabidopsis roots. However, during co-infection of Manduca sexta or Drosophila melanogaster, Ma549's higher blastospore production enhanced its competitive ability within the host. In large M. sexta (fifth instar), blastospores facilitate dispersal, suppress host melanization and prevent Mr2575 from spreading from infection sites, reducing conidia production. However, colonization of smaller hosts such as first instar M. sexta and D. melanogaster did not provide Ma549 with a competitive advantage, as conidial production was dependent on retaining control of the cuticle through which conidiating hyphae emerge. Unexpectedly, Ma549 and Mr2575 segregate within hosts, suggesting resource partitioning with Mr2575 predominating in the thoraxes of Drosophila, especially in females, and Ma549 in the abdomen. In fifth instar M. sexta, Mr2575 was most prevalent around spiracles and the front end of segments, despite Ma549 and Mr2575 having similar susceptibility to hypoxia. Dispersing conidia homogeneously into the hemocoel of fifth instar M. sexta eliminated the blastospore production advantage, making Ma549 and Mr2575 equally competitive, with strict partitioning of Mr2575 at the anterior and Ma549 at the posterior ends of segments. As Metarhizium species have multiple roles in natural ecosystems and agroecosystems these discoveries are relevant to understanding their impact on maintaining biodiversity and for exploiting them to enhance food security. Author summary: Metarhizium spp. are keystone fungi providing essential ecosystem services as plant symbionts and insect pathogens, yet little is known about how different strains interact. We analyzed two strains with different insect-killing strategies: Mr2575 kills insects with toxins before colonizing cadavers, whereas Ma549 rapidly colonizes living insects. Ma549's rapid growth within insects serves as a strong anti-competitive strategy in larger insects. However, in smaller insects, Mr2575 remains competitive in reproductive potential (spore production on cadavers) if it maintains control of the cuticle through which it entered the insect. Consequently, small hosts may be more critical than large hosts for preserving pathogen diversity. Unexpectedly, Mr2575 and Ma549 partition insect hosts between them, promoting coexistence. We also investigated trade-offs between virulence traits and plant colonization traits, finding that Mr2575's rapid growth in sparse nutrients enhances its competitiveness on plant roots. Thus, Mr2575 may be more competitive on plants, while Ma549 excels in large insects, with both strains showing similar competitiveness in small insects. These insights are important for understanding Metarhizium species' roles in biodiversity and their potential to boost food security in natural and agricultural ecosystems. [ABSTRACT FROM AUTHOR]

In recent times, great prominence is moved towards reusing or recycling organic waste into various products with noteworthy applications. In this study, mixed vegetable wastes were decomposed by a microbial consortium composed of bacteria Pseudomonas stutzeri, Bacillus polymyxa, Lactococcus lactis, actinomycetes Streptomyces albus, filamentous fungi Aspergillus niger and yeast strain Candida utilis into high nutrient content humous material. Compost thus obtained was formulated into a liquid growth medium and checked for the mass multiplication of well-known fungal biopesticide Metarhizium anisopliae. Decomposition brought about the production of high nutrient content compost, which revealed high nitrogen (212.3 mg/kg), phosphorous (465.13 mg/kg), potassium (1871.13 mg/kg) and total organic carbon (122.03 mg/kg) content. Liquid medium formulated from the compost recorded maximum biomass, spore production with high germination index of M. anisopliae. Viable spores produced in CBM exhibited noteworthy pesticide activity against the larval instar of gram pod borer Helicoverpa armigera. Formulated medium supported the production of metabolites which inhibited the growth of the human pathogens. These findings demonstrated the noteworthy bioconversion of organic wastes via microbial consortium composting into high growth-promoting, economical, low-cost medium for the mass multiplication of versatile fungal biopesticides and the source of bioactive metabolite production. [ABSTRACT FROM AUTHOR]

The potential of entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae was studied in combination with the commonly used insecticides, viz. clothianidin 50 WDG, imidacloprid 17.8 SL and chlorpyriphos 20 EC, against grubs of Brahmina coriacea (Hope) and Holotrichia longipennis (Blanchard). Clothianidin, imidacloprid and chlorpyriphos were applied at 50, 75 and 100 ppm respectively, and both the fungi were applied at 107 conidia/ml. Synergistic interactions were consistent across all the combinations and calculated values of χ² ranged from 4.2 to 5.73 in both the species of white grubs. The grubs of H. longipennis exhibited a higher degree of synergism than B. coriacea as observed from the mortality data. Among entomopathogenic fungi, M. anisopliae showed more synergism than B. bassiana against both species. This study will also help reduce insecticidal pollution and the cost of interventions for the management of white grubs in a sense that application rates of chemical insecticides and synergists will be reduced. This practice may be incorporated in integrated pest management, and further study of the synergistic interactions will guide how entomopathogenic fungi and insecticides can be combined to combat environmental pollution effectively. [ABSTRACT FROM AUTHOR]

Background/Objective: Fungi that infect insects constitute a very large and diverse group, with approximately 1600 diagnosed and known species belonging to 90 genera, and The fungi have developed their capabilities and ability to infect insects and in different strains, the house fly Musca domestica L. (Diptera: Muscidae), has great medical importance due to its ability to transmit thousands of pathogenic microorganisms such as parasites, fungi, bacteria and viruses. Methods/Statistical analysis: This study was conducted in the insect laboratory at the Faculty of Education for Girls / Anbar University with the aim of isolating and characterizing the most important Entomopathogenic Fungi and evaluating their efficiency on the house fly Musca domestica L. (Diptera: Muscidae). House fly adults were collected and raised in the laboratory, approximately 50 adult house fly insects were collected and treated at three concentrations for seven days in two ways: mixing with food and direct spraying on insects from the suspension [spores] of the fungi Metarhizium anisopliae and Trichoderma harzianum that were isolated from the Zira'a area of Ramadi city. Findings: the results of isolation and diagnosis of fungi showed the registration of the two fungi M. anisopliae and Trichoderma harzianum and their diagnosis was confirmed. (ID: PP035895 and ID: PP065519). The results also showed that there were significant differences between the concentrations used in the experiment, and the percentage of mortality in insects was directly proportional to the increase in the suspension of fungal spores, as the concentration of 1 X 107 spores/ml showed the highest percentage of mortality in the house fly in both methods [direct spraying method and mixing with food], as the percentage of mortality in the direct spraying method from the fungus M. Anisopliae and after a period of 7 days, the percentage of mortality was 85.7% and the percentage of mortality of the same organism by mixing with food was 66.6%, while the same concentration (1 x 107 spores/mL) with the spore suspension of T. harzianum by mixing with food was mortality rate of 85.7% and gave 100% mortality with the direct spraying method after 7 days. The results also revealed that T. harzianum spore suspension at all concentrations used in the experiment showed a higher percentage of lethality than M. anisopliae. Conclusion: The study found that molecular diagnosis is indispensable in the diagnosis of microorganisms, especially fungi because they share many phenotypic and even microscopic characteristics, and the study showed the sensitivity of house fly adults to the spore suspension of the studied fungi, and the study showed the superiority of the fungus T. harzianum in increasing the percentage of mortality compared to M. anisopliae. [ABSTRACT FROM AUTHOR]

The white grubs infests diverse crops of food and horticultural importance such as groundnut, sugarcane, kharif pulses, vegetables areca nut, ginger, potato, etc. and it inflict losses as high as 100 per cent, depending upon the severity of infestation have been uncommon. In present study, attemts have been made to explore the potentiality of entomopathogens for the management of the white grub Holotrichia consanguinea a serious pest in groundnut crop in Rajasthan during kharif seasons, 2018 and 2019. Results revealed that furrow application of Beauveria bassiana, Metarhizium anisopliae powder formulation @ 5 kg (1×1012 CFU/gm) and H. indica galleria @ 5000 galleria per hectare at the time of sowing were significantly superior in reducing the per cent plant mortality (28.00%, 29.33%, 31.77%, respectively) and increase yield over untreated control (12.82, 11.53, 10.78 q/ha respectively). [ABSTRACT FROM AUTHOR]

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The experiment on insecticide evaluation against the Rice Gundhi bug was laid out at Heera Puri research field, Deen Dayal Upadhyaya Gorakhpur University, Gorakhpur, Uttar Pradesh, India in randomized Block Design with three replications and seven treatments including different newer insecticides and phytoextracts viz., imidacloprid 17.8% SL @ 300 ml/ha, NSKE 5% @ 25 l/ha, acetamiprid 20% SP @ 200 g/ha, Neem oil 2% @ 5 l/ha, Metarhizium anisopliae 1 x 109 cfu/ml @ 3 l/ha, thiamethoxam 25% WG @ 200 g/ha, and Untreated plot (Water). The number of bugs after 10 days of the second spray was significantly lower than the untreated plot. However, imidacloprid 17.8 SL @ 300 ml/ha was found superior to the rest of the insecticide treatments, as the lowest number of bugs was recorded by this treatment. The next best treatment was thiamethoxam 25% WG @ 200 g/ha, followed by acetamiprid 20% SP @ 200 g/ha. Neem oil 2% @ 5 l/ha was the least effective treatment. The benefit-cost ratio results revealed that the most cost-effective treatment was imidacloprid 17.8% SL @ 300 ml/ha, whereas the least economical treatment was NSKE 5% @ 25 l/ha. Imidacloprid 17.8 SL @ 300 ml/ha treatment recorded the highest B: C ratio and highest yield as compared to other treatments. [ABSTRACT FROM AUTHOR]

BACKGROUND: An innovative version of the sterile insect technique (SIT) for pest control, called boosted SIT, relies on the use of sterile males coated with a biocide to control a target wild pest population of the same species. The objective of the present study was to assess the relevance of such technology to control the fruit fly Bactrocera dorsalis and fruit losses in mango orchards using. An agent‐based simulation model named BOOSTIT was used to explore the reduction of fruit losses thank to sterile male fruit flies control and economic benefits according to different strategies of sterile male release. The simulation considered a landscape of 30.25 ha made up of four mango orchards. RESULTS: The SIT and the boosted SIT reduced fruit losses when releases were made before the mango fruiting period. According to model simulations, releases should be performed at least seven times at 2‐week intervals and with a sterile/wild male ratio of at least 10:1. Considering the benefit/cost ratio (BCR), few releases should be done with a late start date. The BCR showed economic gains from the two control methods, the number of saved fruits and BCR being higher for SIT. CONCLUSION: Our simulations showed that SIT would have better results than the boosted SIT to contribute to an effective control of Bactrocera dorsalis at the scale of a small landscape. We highlight the need for laboratory studies of other types of pathogen to find a suitable one with higher incubation time and lower cost. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The present study revealed the consequences of the interaction of a widely used bioinsecticide and endophyte Metarhizium anisopliae with the hazardous mycotoxin zearalenone (ZEN) as a pure substance and with ZEN as a native component of a crude Fusarium extract. In the environment, microorganisms encounter a mixture of metabolites secreted by other organisms living in the same area, not single substances. The obtained results suggest that M. anisopliae, exposed to a variety of active substances produced by Fusarium graminearum, is able to eliminate ZEN. Within 14 days, M. anisopliae biotransformed 90.8% and 85.8% of ZEN as a pure substance and ZEN as a native component of the F. graminearum extract from Rice Medium (E-Fg-RM), respectively, through reduction predominantly to α-epimers of zearalenols and zearalanols, considered more estrogenic than ZEN, which can raise concerns. Compared to pure ZEN, E-Fg-RM significantly affected the production of Metarhizium secondary metabolites by increasing the destruxins amount by approximately 20–25% and reducing the swainsonine content by 96.2%. All these findings provide a possible picture of the interaction of M. anisopliae with ZEN in the wild, mainly as a result of the use of crude extract from Fusarium, which contained a mixture of different metabolites. [ABSTRACT FROM AUTHOR]

Fall armyworm (FAW), Spodoptera frugiperda is a generalist pest known to feed on more than 300 plant species, including major staple crops such as rice, maize and sorghum. Biological control of FAW using a combination of a major indigenous egg parasitoid Telenomus remus and entomopathogenic fungi was explored in this study. Metarhizium anisopliae strains (ICIPE 7, ICIPE 41, and ICIPE 78) and Beauveria bassiana ICIPE 621 which demonstrated effectiveness to combat the pest, were evaluated through direct and indirect fungal infection to assess their pathogenicity and virulence against T. remus adults, S. frugiperda eggs and their effects on T. remus parasitism rates. Metarhizium anisopliae ICIPE 7 and ICIPE 78 exhibited the highest virulence against T. remus adults with LT50 values >2 days. ICIPE 7 induced the highest T. remus mortality rate (81.40 ± 4.17%) following direct infection with dry conidia. Direct fungal infection also had a significant impact on parasitoid emergence, with the highest emergence rate recorded in the M. anisopliae ICIPE 7 treatment (42.50 ± 5.55%), compared to the control ± (83.25 ± 5.94%). In the indirect infection, the highest concentration of 1 x 109 conidia ml-1 of ICIPE 78 induced the highest mortality (100 ± 0.00%) of T. remus adults, and the highest mortality (51.25%) of FAW eggs, whereas the least FAW egg mortality (15.25%) was recorded in the lowest concentration 1 x 105 conidia ml-1 of ICIPE 41. The number of parasitoids that emerged and their sex ratios were not affected by the different fungal strain concentrations except in ICIPE 7 at high dose. This study showed that potential combination of both M. anisopliae and B. bassiana with T. remus parasitoid can effectively suppress FAW populations. [ABSTRACT FROM AUTHOR]

Metarhizium anisopliae (Clavicipitaceae, Hypocreales) is a globally distributed entomopathogenic fungus, which has been largely studied and used in agriculture for its potent entomopathogenicity. Since its taxonomic establishment as a member of Metarhizium, many closely related taxa have been described with highly similar morphology (cryptic species). A holotype specimen of M. anisopliae is not extant, and the ex-neotype strain (CBS 130.71) does not form a monophyletic clade with other strains, up to now, recognized as M. anisopliae sensu stricto. In this study, we have conducted an integrative taxonomic treatment of M. anisopliae sensu lato by including the ex-neotype strain of M. anisopliae, other unknown strains from our collections identified as M. anisopliae s. lat., as well as other known species that have been previously delimited as closely related but distinct to M. anisopliae. By including whole-genome sequencing, morphometric analysis, LC–MS based metabolomics, and virulence assays, we have demonstrated that M. anisopliae s. str. should also include M. lepidiotae (synonym), and that M. anisopliae s. str. differentiates from the other species of the complex by its metabolome and less severe entomopathogenicity. New taxa, namely M. hybridum, M. neoanisopliae and M. parapingshaense spp. nov., are proposed. The novel taxa proposed here have strong phylogenomics support, corroborated by fine-scale differences in the length/width of conidia/phialides, while the metabolomics and virulence data still largely overlap. We have also demonstrated via population genomics data the existence of local clonal lineages, particularly the one corresponding to the persistence of a biocontrol candidate strain that has been used in the field application for three years. This study showcases the utility of combining various data sources for accurate delimitation of species within an important group of fungal biocontrol agents against pest insects. [ABSTRACT FROM AUTHOR]

Opisthoplatia orientalis Burmeister is a traditional medicinal insect in China that is mass‐produced by entrepreneur farmers. However, low temperatures pose a serious threat to the survival and quality of this insect, causing economic losses to the farmers. This study aimed to test the hypothesis that low temperatures induce a trade‐off between immunity and physical condition of O. orientalis. The phenoloxidase activity and hemocyte concentration of O. orientalis under 25°C and 5°C were measured by micro‐determination and haemocytometre, respectively. The immunocompetence of the insect was also evaluated by inoculating it with Metarhizium anisopliae M11, an entomopathogenic fungus. The physical factors (protein content, fatty acid content and body weight) of O. orientalis were determined accordingly. The results showed that phenoloxidase activity and hemocyte concentration increased from 9.21 mOD/min and 122.30 hemocyte/ml to 3.64 mOD/min and 55.70 hemocyte/ml when temperature down‐regulated from 25 to 5°C. However, the insects at 5°C had higher protein and fatty acid content, but lower body weight, than those at 25°C. Moreover, there was a significant negative correlation between immunity and protein or fatty acid content, indicating a trade‐off between immunity and physical condition in O. orientalis under low temperatures. This study provides useful insights for improving the efficiency of insect breeding and enhancing the quality of this medicinal insect. [ABSTRACT FROM AUTHOR]

In this study, a powder of the larvae, pupae and adults of the fall armyworm Spodoptera frugiperda (J.E. Smith) was obtained, and 1% of this powder was added to solid culture media potato dextrose agar (PDA) and liquid media Sabouraud dextrose broth (SDB) and minimal medium (MM), to induce the development and virulence of the M1 strain of Metarhizium anisopliae (Metchnikoff) Sorokin, which is pathogenic against the L1 larvae of S. frugiperda. The pupa powder was selected because it causes the greatest degree of radial growth of the M1 strain (73.3 mm) in PDA (p < 0.05). The enzymatic activity of M1, in the medium with pupa powder of S. frugiperda (I), was increased on day 4 for the proteases in MMI (1.1 U min-1) and on day 2 for the lipases in MMI (7034 U) and SDBI (6004 U) (p<0.05). In the pathogenicity bioassay, the M1 strain grown on PDA with the pupa powder caused 90% mortality on day 4 in the L1 larvae, while M1 grown on PDA without the powder caused 50%. With the pupa powder, the growth of the M1 strain increased, which promoted the production of enzymes, thus, increasing the pathogenicity of M. anisopliae against the L1 larvae of S. frugiperda. [ABSTRACT FROM AUTHOR]

(1) Background: Insect pathogenic fungi of the genus Metarhizium are under study and in application as highly solicited, more eco-system friendly substitutes for chemical insecticides in many countries and in different agricultural contexts. In Cuba and Florida, Metarhizium strains have previously been isolated from economically important coffee and sugar cane pests. (2) Methods: Unambiguous species delineation within the Metarhizium anisopliae species complex is methodologically challenging. Recently, a species-discriminating PCR approach has been developed based on ribosomal intergenic spacer (rIGS) sequences that covered the prominent four "PARB" species within the complex. This approach is combined here with further genetic markers and is extended to a further species. (3) Results: Metarhizium isolates from Cuba, found to be more naturally associated with the coffee berry borer, Hypothenemus hampei, were morphologically, microscopically and molecular taxonomically characterized. Multilocus sequence analysis based on 5TEF, MzIGS3 and rIGS markers delineated these weevil-associated strains from all previously established Metarhizium species. (4) Conclusions: The isolates under study represent a new fungal taxon proposed to be designated Metarhizium caribense. The rIGS-based species-discriminating diagnostic PCR is a suitable tool for the identification of new Metarhizium species and can be productively combined to approaches using further genetic markers. [ABSTRACT FROM AUTHOR]

The use of entomopathogenic fungi (EF) is a promising alternative for the control of Rhipicephalus microplus, an important tick affecting cattle globally. This study aimed to evaluate the effect of ultraviolet irradiation (UV) exposure on the percentage of conidia germination and number of colony-forming units of eight strains of Metarhizium anisopliae (MaV55, MaV35, MaV31, MaV25, MaV13, Ma08, MaV05, and MaV02). The UV (UV-A and UV-A+B) irradiation was carried out with an ultraviolet radiation emission lamp. The conidia of each strain were exposed to the UV irradiation treatments for 3 h. MaV25, MaV08, MaV05, MaV13, and MaV31 showed higher tolerance to UV-A radiation exposure, as assessed by conidia germination. UV-A+B radiation decreased the germination percentage of all the M. anisopliae strains. The eight evaluated strains showed good tolerance to UV-A radiation, as assessed by the development of colony-forming units (CFU). UV-A+B radiation did not significantly affect (p > 0.05) the count of the CFU of six of the M. anisopliae strains evaluated (MaV35, MaV13, MaV08, MaV05, MaV31, and MaV02). The novel findings of the UV-tolerant M. anisopliae strains may potentially improve the effectiveness of EF under environmental conditions. Integral research under real tropical conditions is advised to evaluate the effectiveness of the EF strains. [ABSTRACT FROM AUTHOR]

The coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Scolytidae), is one of the most important coffee pests, and is present in almost all countries producing this crop, causing annual losses of US$350 million. Natural factors regulate H. hampei populations in coffee crops. Ecological life table analysis is a robust tool that allows the estimation of the mortality caused by these factors. Therefore, the main objective of this study was to determine the critical stage and the key mortality factors affecting H. hampei in the field. Data for constructing life tables were collected over 2 years in six coffee plantations in the regions of Paula Cândido and Viçosa, Minas Gerais State, Brazil. The average mortality of H. hampei was 75.34% ± 5.85%. The mortality at each developmental stage was 24.32% ± 2.83% (eggs), 7.29% ± 1.01% (first instar), 11.58% ± 1.46% (second instar), 9.68% ± 1.10% (third instar), 7.45% ± 1.01% (pupa), and 15.02% ± 1.29% (adult; n = 124). The mortality factors observed and quantified in this study were parasitism by Prorops nasuta (Hymenoptera: Bethylidae), predation by ants (Crematogaster spp., Pheidole spp. and Solenopsis spp.), physiological disorders and fungal infection by Beauveria bassiana and Metarhizium anisopliae. The critical mortality stage of H. hampei was the adult stage. The key factors for mortality of H. hampei adults were predatory ants, followed by B. bassiana and malformations (these insects had deformities in the head, wings, legs, or abdomen). This information is essential to conserve the activities of natural enemies and, thus maintain H. hampei natural mortality factors in coffee plantations, reducing damage to the crops and the need for excessive insecticide interventions. [ABSTRACT FROM AUTHOR]

The pathogenicity of the fungus, Metarhizium anisopliae, commonly referred to as the green muscardine fungus, has been demonstrated against numerous insect species inhabiting various habitats. Laboratory experiments were carried out to investigate the impact of naturally derived adjuvants on the growth and spore characteristics of M. anisopliae. The natural polysaccharide (guar gum, gum acacia and carboxymethyl cellulose), vegetable oil (groundnut and cottonseed oil) and a synthetic adjuvant were used in the study at three distinct concentrations: 0.05%, 0.1%, and 0.2% v/v or w/v. Among the various adjuvants tested, both gum acacia and guar gum at concentrations of 0.1% and 0.2% were observed to have a positive impact on the growth of the fungus, resulting in enhanced radial mycelial growth, conidiospore production, surface area coverage, and conidiospore germination. Conversely, this fundamental study highlights the detrimental effects of synthetic adjuvants on the growth and spore characteristics of M. anisopliae. [ABSTRACT FROM AUTHOR]

The present study was undertaken to screen the pesticidal activity of extra cellular metabolites derived from entomopathogenic fungus Metarhizium anisopliae against Spodoptera litura and Aphis crassivora an important aphid pest under laboratory and pot culture condition. Fungal strain used in the study was isolated from the soil by serial dilution technique. Isolated fungal strain was mass multiplied in a liquid medium sabouraud maltose yeast extract broth under optimum condition (28°C). Culture filtrate and the collected culture derived from the medium was used for the source of metabolites. The culture filtrate was extracted with ethyl acetate, acetone and the concentrated extract was screened for pesticidal activity against Spodoptera litura and Aphis crassivora adopting leaf disc and pot culture assay respectively. Extracted metabolites showed pesticidal activity against both insect pests. However, notable effect was recorded in ethyl acetate derived metabolites. The ethyl acetate extract treatment recorded 91.2% mortality at a concentration of 100 μg against Spodoptera litura and 100% mortality at a concentration of 100 μg against Aphis crassivora. Phytotoxicity was determined by measuring the germination index (GI) of groundnut seeds by exposing it to the crude metabolites extracted in ethyl acetate and acetone. The results showed that the germination index was high on ethyl acetate extract which was recorded to be 98.0% and germination index of acetone extract was found to be 90.0%. Production of insecticidal metabolites from M. anisopliae under laboratory condition proposed the possible utilization of metabolites as an effective, safe pesticidal agent against economic important insect pests. No impact on the seedling emergence revealed non phytotoxic effect of the metabolites. [ABSTRACT FROM AUTHOR]

BACKGROUND: Xylella fastidiosa is an important causative agent of Olive Quick Decline Syndrome in the Apulia region of Italy. The current study evaluated the bioefficacy of three entomopathogenic fungal strains: Beauveria bassiana SGB7004, Metarhizium robertsii SGB1K, and Akanthomyces lecanii SGB4711 against Philaenus spumarius the main vector of this pathogen, under laboratory conditions. Pathogenicity bioassays were performed by dipping nymphs and adults of P. spumarius in an aqueous suspension of powdered fungal culture (PFC) or conidial suspension (CS) of the three fungal strains. RESULTS: Both B. bassiana SGB7004 and M. robertsii SGB1K affected the viability of nymphs, resulting in more than 80% mortality at 48 h post treatment, while the effect of A. lecanii SGB4711 was not statistically significant. On adults, all three biocontrol strains were effective in a time‐ and concentration‐dependent manner. The PFCs of B. bassiana SGB7004, M. robertsii SGB1K, and A. lecanii SGB4711 at the highest concentration tested (120 mg mL−1) resulted in 97%, 83% and 27% mortality at the trial endpoint (120 h), respectively. Mycelial growth was observed on 38.5%, 37.0% and 61.5% of dead insects treated with B. bassiana SGB7004 (2.3 × 108 CFU mL−1), M. robertsii SGB1K (3.8 × 106 CFU mL−1) and A. lecanii SGB4711 (5.4 × 108 CFU mL−1), respectively. None of the PFCs of the tested strains was pathogenic when injected into nymph spittle. CONCLUSIONS: Beauveria bassiana SGB7004 and M. robertsii SGB1K significantly affected the survival of P. spumarius nymphs and adults, while A. lecanii SGB4711 was not effective on nymphs and only slightly effective against adults. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

BACKGROUND: Most studies on efficacy of fungal pathogens and predatory mites against Tetranychus urticae have been done on individual species in the laboratory. We evaluated fungi and predatory mites separately and together against glasshouse populations of T. urticae on chrysanthemum plants. First, effectiveness of the fungal pathogens Beauveria bassiana (Bb88) and Metarhizium anisopliae (Ma129) was compared; then, effectiveness of the predatory mites Phytoseiulus persimilis and Neoseiulus californicus. Based on the results, N. californicus and isolate Ma129 were selected and evaluated in combination. In all experiments, treatment effects were assessed for eggs and motile stages of T. urticae. RESULTS: The first experiment showed no significant effect of either fungal isolate on T. urticae populations, except on plants initially infested with 20 mites, where more eggs were found in the control compared to the fungal treatments. In the second experiment, both predatory mites were equally effective at reducing T. urticae populations compared with the control, regardless of initial T. urticae population density. The last experiment demonstrated that populations of T. urticae were reduced most when M. anisopliae (Ma129) and N. californicus were applied together, compared with the control and when each natural enemy was applied separately. CONCLUSIONS: Metarhizium anisopliae (Ma129) and B. bassiana (Bb88) isolates did not have a significant effect on reducing T. urticae populations. Both predatory mites reduced T. urticae populations, regardless of T. urticae density. Combined application of M. anisopliae (Ma129) and N. californicus were more effective against T. urticae than the control or when each agent was applied separately. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Abstract Background Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae) infestation poses a serious risk to bitter gourd cultivation. Traditionally, B. cucurbitae has been controlled using synthetic pesticides, which have drawbacks such as non-target toxicity and pest resistance. Entomopathogenic fungi (EPF) provide concentrated ecological alternatives, which support ongoing pest reduction and sustainable agriculture by adhering to Integrated Pest Management principles. Therefore, EPF provides a viable alternative for chemical control of B. cucurbitae, addressing its shortcomings and promoting environmentally friendly pest control technology. This study evaluated the effectiveness of entomo-vectored horizontal transmission devices (EV-HTD) against B. cucurbitae in bitter gourd fields, focusing on GF-120 and Butanone acetate. Assessment parameters include converting fruit infestation data into yield loss per plant, marketable fruit yield per plant, marketable yield per hectare, and yield loss per hectare. Results The highest mean percentage of entomo-vectored B. cucurbitae (70.50%) was found in plots treated with Butanone acetate + B. bassiana-based EV-HTD. This was followed by GF-120 + B. bassiana-based EV-HTD (66.18%), Butanone acetate + M. anisopliae-based EV-HTD (58.95%), and GF-120 + M. anisopliae-based EV-HTD (54.78%). The Butanone acetate + B. bassiana-based EV-HTD produced the highest mean number of spores per B. cucurbitae (7.80 spores/cm2), while the other treatments produced low spore counts. Plots treated with Butanone acetate + B. bassiana-based EV-HTD had the highest percentage mortality of B. cucurbitae (81.20%). The percentage of fruit infestation varied between 9.00 and 34.00%, with the least amount of infestation seen in plots treated with B. bassiana + Butanone acetate. There were minimal yield losses in Butanone acetate. The Butanone acetate + B. bassiana-based EV-HTD showed the lowest yield losses (66.66 g/plant), while the other treatments showed high losses. Plots treated with Butanone acetate + B. bassiana-based EV-HTD had the highest marketable yield per plant (673.87 g/plant), while yields in control treatments were low. Plots treated with Butanone acetate + B. bassiana-based EV-HTD had the highest marketable yield (2217.85 kg/ha). Lastly, plots treated with Butanone acetate + B. bassiana-based EV-HTD (219.40 kg/ha) showed the lowest yield losses per hectare. Conclusions According to the study’s findings, Butanone acetate-based EV-HTD was more successful than GF-120. Furthermore, B. bassiana was more effective at controlling B. cucurbitae than M. anisopliae. With a maximum cost–benefit ratio of 14.99, the treatment Butanone acetate + B. bassiana was shown to be the most advantageous economically, suggesting its potential for use in practical pest management techniques.

Entomopathogenic fungi (EPF) represent the future of pest control, as they have the capacity to induce serious infections in insects, trigger epizootics, and effectively manage insect populations. The study aimed to identify and test indigenous EPF isolates against Tribolium confusum using soil samples from Northwestern and central Saudi Arabia. Galleria mellonella larvae were used as insect bait to collect EPF from soil samples from date palm groves, fruit groves, and maise fields. Only 27.3% of soil samples tested positive for palm grove-originating EPFs. Polymerase chain reaction (PCR) identified the cultured fungi as Metarhizium anisopliae and Beauveria bassiana. Tribolium confusum pathogenicity was assessed using nine local fungal isolates. Different fungus isolates were tested for their pathogenicity on larval and adult T. confusum. The results showed that M. anisopliae MaSA-2, MaSA-3, and MaSA-4 had infection rates of 85%, 85%, and 75%, respectively, in last-instar T. confusum larvae and 5%, 10%, and 20% in adults within seven days. On the other hand, B. bassiana BbSA-4 showed a 100% infection rate in T. confusum larvae, while BbSA-5, BbSA-6, BbSA-7, BbSA-8, and BbSA-9 had infection rates of 70%, 80%, 80%, 75%, and 95%, respectively. In terms of adult mortality rates, B. bassiana BbSA-4, BbSA-5, BbSA-6, BbSA-7, BbSA-8, and BbSA-9 had rates of 25%, 35%, 35%, 20%, 20%, and 40%, respectively. The research findings show that indigenous B. bassiana and M. anisopliae fungus isolates successfully eliminated T. confusum larvae, although their effectiveness against adults was limited.

The research aimed to isolate and characterize local entomopathogenic fungi from different agricultural ecosystems and assess their pathogenicity against cowpea aphid. Forty soil samples were collected from different ecosystems. In total, 38 fungal isolates were collected from the cadavers' exteriors of Galleria mellonella larvae. Microscopic observation of these fungal isolates revealed that they belonged to nine fungal genera among them Metarhizium sp. (3 isolates) and Beauveria sp. (5 isolates). Primary experiments showed different bioactivities of the isolates of the two selected genera against the target insect. Only Metarhizium isolates (M3 & M5) and Beauveria isolates (B1 & B5) were selected for further investigations. Identification of fungal cultures by morphological characters, molecular characterization, ITS sequencing, and NCBI Blast showed that two isolates were confirmed as M. anisopliae and two isolates as B. bassiana. The bioassay evaluation revealed that M. anisopliae (M3 and M5) and B. bassiana (B1) exhibited significant virulence against A. craccivora, as evidenced by their high LT50 and LC50 values and substantial sporulation on the cadavers. Therefore, they could be used for the formulation of pesticides targeting this pest on cowpea and other vegetable crops.

Penelitian ini bertujuan untuk mengetahui efektivitas tiga isolat jamur M. anisopliae dan ekstrak tembakau dengan kerapatan dan konsentrasi berbeda dalam mengendalikan populasi hama P. xylostella. Penelitian ini dilaksanakan pada bulan Oktober sampai Desember 2022 di Laboratorium Agens Hayati Brenjonk dan Laboratorium BPTPH Pasuruan. Penelitian ini menggunakan Rancangan Acak Kelompok (RAK) dengan 16 perlakuan dan 4 kali ulangan. Setiap perlakuan menggunakan 15 ekor P. xylostella yang diuji untuk mengetahui mortalitas, daya hambat makan, LC50, dan LT50 dengan sidik ragam ANOVA dan analisis probit. Hasil penelitian menunjukkan bahwa mortalitas dan daya hambat tertinggi diperoleh dari isolat jamur koleksi Bojonegoro dan Jatisari dengan kerapatan 108 serta ekstrak tembakau konsentrasi 50 ml/L. Sedangkan nilai LC50 terbaik terdapat pada isolat jamur koleksi Jatisari yaitu 4,99% dan LT50 terbaik terdapat pada isolat jamur koleksi Bojonegoro yaitu 133,09 jam. Berdasarkan hasil penelitian menunjukkan bahwa ketiga isolat jamur M. anisopliae dan ekstrak tembakau memiliki kemampuan terbaik dalam menekan populasi hama P. xylostella.

Abstract Background Green bean, Phaseolus vulgaris L. (Fam.: Leguminosae) is a widely consumed grain legume prized for its edible seeds and pods. It is susceptible to infestations with various pests as insects and mites throughout the growing season. In this study, the efficacy of the predatory species, Chrysoperla carnea (Stephens), Phytoseiulus persimilis Athias-Henriot and the entomopathogenic fungus (EPF) Metarhizium anisopliae (Hypocreales: Clavicipitaceae), as well as conventional pesticides, Mospilan and Vertimec, were evaluated against the most important pests, mainly the whitefly, Bemisia tabaci (Genn.) and the two-spotted spider mite, Tetranychus urticae Koch, infesting the green beans cultivated at two locations Giza and El-Menoufia Governorates in Egypt. Results The findings demonstrate that treatments using C. carnea and M. anisopliae effectively reduced the whitefly population, while pesticide treatments were comparatively less effective. In the Giza plots, at the end of the experiments, the use of both M. anisopliae and C. carnea showed high reductions in whitefly population (85.57 and 84.87%), respectively, while in El-Menoufia, C. carnea (97.74%) was the most effective treatment followed by M. anisopliae (90.32%). Pesticide treatment in this case yielded a reduction rate of (22.76 and 59.67%) in Giza and El-Menoufia plots, respectively. However, for spider mite control, P. persimilis proved to be the most effective treatment in Giza and El-Menoufia plots, reducing the spider mite population to 98.44 and 96.14%, respectively. Metarhizium anisopliae treatment also displayed moderate effectiveness, with reduction rates of 75.62 and 75.37% in Giza and El-Menoufia plots, respectively. In comparison, pesticide treatment showed low effectiveness, with reduction rates of only 23.92 and 53.16% in the two locations, respectively. Conclusion Applications of the predator, C. carnea and the EPF, M. anisopliae were highly effective in reducing the population of whitefly, while the predator mite P. persimilis proved to be the most effective for controlling the spider mites. Overall, the study suggests that biocontrol agents, such as the predators and the EPF, can be considered as alternatives to synthetic chemical pesticides for controlling pests infesting green beans.

AbstractLeopard moth Zeuzera multistrigata (Lepidoptera: Cossidae) is a xylophagous insect whose larval stages are cryptic woodborers in apple and many other horticultural and forestry crops. The present study was carried out to test effectiveness of some biocontrol agents along with most effective pesticide against this pest on apple (cv. Royal Delicious) in Himachal Pradesh, India, for three consecutive years by injecting biocontrol agents/chemicals directly into galleries made by pest larvae. Investigations were conducted to observe the effect of Beauveria bassiana, Metarhizium anisopliae, Steinernemma feltiae, Heterorhabditis bacteriophora and azadirachtin along with a synthetic pesticide chlorpyriphos. The application of chlorpyriphos (0.04%) caused a 100%%larval mortality, whereas among the biocontrol agents H. bacteriophora (5000 IJs/gallery) exhibited the highest mortality of 66.7 to 80.0%% during these three years, which was significantly at par with application of azadirachtin (Neem baan) 1500 ppm @ 2 ml/Litre and S. feltiae @ 5000 IJs/gallery. The lowest mortality (33.3%) was observed with B. bassiana (5 g/Litre) for the first two years and with M. anisopliae (5 g/L) in the third year i.e. 40.0%. Application of entomopathogenic nematode H. bacteriophora (5000 IJs/gallery) is proven to be best among all biocontrol agents. [ABSTRACT FROM AUTHOR]

Background: Lyme disease continues to expand in Canada and the USA and no single intervention is likely to curb the epidemic. Methods: We propose a platform to quantitatively assess the effectiveness of a subset of Ixodes scapularis tick management approaches. The platform allows us to assess the impact of different control treatments, conducted either individually (single interventions) or in combination (combined efforts), with varying timings and durations. Interventions include three low environmental toxicity measures in differing combinations, namely reductions in white-tailed deer (Odocoileus virginianus) populations, broadcast area-application of the entomopathogenic fungus Metarhizium anisopliae, and fipronil-based rodent-targeted bait boxes. To assess the impact of these control efforts, we calibrated a process-based mathematical model to data collected from residential properties in the town of Redding, southwestern Connecticut, where an integrated tick management program to reduce I.xodes scapularis nymphs was conducted from 2013 through 2016. We estimated parameters mechanistically for each of the three treatments, simulated multiple combinations and timings of interventions, and computed the resulting percent reduction of the nymphal peak and of the area under the phenology curve. Results: Simulation outputs suggest that the three-treatment combination and the bait boxes–deer reduction combination had the overall highest impacts on suppressing I. scapularis nymphs. All (single or combined) interventions were more efficacious when implemented for a higher number of years. When implemented for at least 4 years, most interventions (except the single application of the entomopathogenic fungus) were predicted to strongly reduce the nymphal peak compared with the no intervention scenario. Finally, we determined the optimal period to apply the entomopathogenic fungus in residential yards, depending on the number of applications. Conclusions: Computer simulation is a powerful tool to identify the optimal deployment of individual and combined tick management approaches, which can synergistically contribute to short-to-long-term, costeffective, and sustainable control of tick-borne diseases in integrated tick management (ITM) interventions. [ABSTRACT FROM AUTHOR]

Cauliflower is an essential vegetable crop globally, providing essential nutrition and income for millions of people. However, it is highly susceptible to various pests, resulting in a significant reduction in both quality and yield. The conventional solution to control these pests has been the use of chemical pesticides, but their usage has resulted in severe negative effects on the environment, human health, and non-target organisms. Therefore, it is crucial to explore alternative methods, such as biological control with microbials, to manage cauliflower pests sustainably. The study aimed to assess the effectiveness of different microbial agents for controlling Pieris brassicae and Plutella xylostella on cauliflower over two rabi seasons of 2020–2021 and 2021–2022. The microbial agents evaluated were Beauveria bassiana 1.0% WP at 1.50 and 1.75 kg/acre, Bacillus thuringiensis var kurstaki 0.5% WP at 600 g and 800 g/acre, Metarhizium anisopliae 1.0% WP at 1.50 and 1.75 kg/acre, and Spinosad 2.5 SC at 240 ml/acre. The results showed that B. thuringiensis var kurstaki 0.5% WP (800 and 600 g/acre) was the most effective treatment against P. brassicae and P. xylostella on cauliflower, followed by B. bassiana 1.0% WP (1.75 g/acre). These two treatments significantly reduced the population of the target pests on cauliflower crops compared to untreated control. Additionally, treatments with B. bassiana and M. anisopliae recorded the highest populations of natural enemies, including coccinellid beetles, spiders, syrphids, and Cotesia glomerate cocoon on cauliflower. Regarding yield, spinosad 2.5 SC was found to be at par with B. thuringiensis var kurstaki 0.5% WP (800 and 600 g/acre) during rabi seasons of 2020–2021 and 2021–2022. The study suggests that B. thuringiensis var kurstaki 0.5% WP (800 and 600 g/acre) can be a suitable alternative to chemical pesticides, against P. brassicae and P. xylostella on cauliflower. [ABSTRACT FROM AUTHOR]

Looper caterpillars including the Biston (= Buzura) suppressaria (Lepidoptera: Geometridae) and Hyposidra talaca (Lepidoptera: Geometridae), are recorded as major defoliating pests in tea [Camellia sinensis (L.) O. Kuntze], are responsible for a decline in tea bush quality, a decrease in yield, and extensive foliage deterioration. The hasty and unlawful usage of chemical pesticides for looper control has adverse effects, such as the deterioration of natural soil fertility, disturbance of beneficial plant-microbe interactions, and an elevated risk of pesticide resistance. Undesired pesticide residues are frequently detected in the final tea product as pesticides have been applied to eradicate looper caterpillars in tea crop. In the current investigation, spore suspensions of two entomopathogenic fungi, Beauveria bassiana (MG547581) and Metarhizium anisopliae (MG547580), have been employed in vitro to evaluate their effectiveness in controlling three distinct stages of the life cycle of looper caterpillar. The highest rates of microbial infection and pest mortality were measured in the prepupal stage of each pest during in vitro incubation studies and bioassay. Pest mortality up to 90% was recorded with a spore load of 3 × 107 cfu/mL at the prepupal stage, while mortality in adults was up to 70% and influenced by species variations. The lowest median lethal dose (LD50) values of M. anisopliae were recorded as 1.1 × 106 (prepupae), 1.9 × 106 (pupae) and 3.2 × 106 (adult) against H. talaca and against B. suppressaria both the entomopathogens showed equal LD50 values in all distinct stages like prepupae (5.3 × 105), pupae (3.2 × 106) and adult (7.6 × 104). Further, median lethal time (LT50) values increased gradually from prepupae, and pupae to adult, both in cases of H. talaca (3.69, 3.97 and 4.40 days in 3 × 107 cfu/mL of M. anisopliae) and B. suppressaria (4.30, 4.72 and 5.34 days in 3 × 107 cfu/mL of B. bassiana) due to entomopathogenesis. As eco-friendly and nonchemical options for tea looper caterpillar management appear to be limited, the current study explores putative microbial alternatives such as B. bassiana and M. anisopliae for the sustainable management of two significant defoliating pests of tea. [ABSTRACT FROM AUTHOR]

Sustainable control of cereal aphids during late spring and early summer requires alternatives to mitigate increasing resistance to chemical insecticides. Emulsifiable oil-formulated Beauveria bassiana ZJU435 (Bb) and Metarhizium anisopliae CQ421 (Ma) commercially registered in China were tested as alternatives against the aphids Rhopalosiphum padi and Sitobion avenae, which infested 29% of winter wheat tillers with ~11 aphids per tiller at elongating stage, followed by rapid population growth. Bb and Ma were sprayed biweekly at the low and high rates of 1.0 × 1013 and 1.5 × 1013 conidia ha−1 over five weeks of three independent field trials from elongating to milk ripening stages. In four fungal treatments (three randomized plots of 100 m2 per treatment), percent tiller infestation and aphid density were reduced respectively to 19–23% and 9.5–15.8 aphids per tiller, contrasting with 53% and 63 (~110 at peak) aphids per tiller in unsprayed control throughout the field trials. Overall mean (±SE) control efficacies of Bb and Ma assessed with weekly sampled aphid densities were 85% (±0.3) and 80% (±0.6) at the high rate and 79% (±0.6) and 75% (±0.8) at the low rate, and differed significantly with each other at either spray rate (P < 0.01). Conclusively, both Bb and Ma were highly efficacious against cereal aphids, and Bb was superior to Ma perhaps due to a lower rate of larger Ma conidia deposited on wheat plants at either spray rate. Their high efficacies were attributed to the qualified formulation and the season warming up with frequent drizzles to favor fungal aphidicidal activity. [ABSTRACT FROM AUTHOR]

Background: Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae) infestation poses a serious risk to bitter gourd cultivation. Traditionally, B. cucurbitae has been controlled using synthetic pesticides, which have drawbacks such as non-target toxicity and pest resistance. Entomopathogenic fungi (EPF) provide concentrated ecological alternatives, which support ongoing pest reduction and sustainable agriculture by adhering to Integrated Pest Management principles. Therefore, EPF provides a viable alternative for chemical control of B. cucurbitae, addressing its shortcomings and promoting environmentally friendly pest control technology. This study evaluated the effectiveness of entomo-vectored horizontal transmission devices (EV-HTD) against B. cucurbitae in bitter gourd fields, focusing on GF-120 and Butanone acetate. Assessment parameters include converting fruit infestation data into yield loss per plant, marketable fruit yield per plant, marketable yield per hectare, and yield loss per hectare. Results: The highest mean percentage of entomo-vectored B. cucurbitae (70.50%) was found in plots treated with Butanone acetate + B. bassiana-based EV-HTD. This was followed by GF-120 + B. bassiana-based EV-HTD (66.18%), Butanone acetate + M. anisopliae-based EV-HTD (58.95%), and GF-120 + M. anisopliae-based EV-HTD (54.78%). The Butanone acetate + B. bassiana-based EV-HTD produced the highest mean number of spores per B. cucurbitae (7.80 spores/cm2), while the other treatments produced low spore counts. Plots treated with Butanone acetate + B. bassiana-based EV-HTD had the highest percentage mortality of B. cucurbitae (81.20%). The percentage of fruit infestation varied between 9.00 and 34.00%, with the least amount of infestation seen in plots treated with B. bassiana + Butanone acetate. There were minimal yield losses in Butanone acetate. The Butanone acetate + B. bassiana-based EV-HTD showed the lowest yield losses (66.66 g/plant), while the other treatments showed high losses. Plots treated with Butanone acetate + B. bassiana-based EV-HTD had the highest marketable yield per plant (673.87 g/plant), while yields in control treatments were low. Plots treated with Butanone acetate + B. bassiana-based EV-HTD had the highest marketable yield (2217.85 kg/ha). Lastly, plots treated with Butanone acetate + B. bassiana-based EV-HTD (219.40 kg/ha) showed the lowest yield losses per hectare. Conclusions: According to the study's findings, Butanone acetate-based EV-HTD was more successful than GF-120. Furthermore, B. bassiana was more effective at controlling B. cucurbitae than M. anisopliae. With a maximum cost–benefit ratio of 14.99, the treatment Butanone acetate + B. bassiana was shown to be the most advantageous economically, suggesting its potential for use in practical pest management techniques. [ABSTRACT FROM AUTHOR]

This study focused on evaluating the effectiveness of seed treatments and different sugar beet varieties in controlling flea beetles (Chaetocnema tibialis) and sugar beet weevils (Asproparthenis punctiventris) in Croatia. The field trials were conducted in Vukovar-Sirmia County and targeted the developmental stages of sugar beet from BBCH 12 to BBCH 31. Although the sowing was done within the optimal period, no clear pattern between germination of the seeds and susceptibility was identified as the results showed different responses at different development stages and among the three variants. The experimental design comprised no insecticide, thiamethoxam + tefluthrin, cyantraniliprole, flupyradifurone and Beauveria bassiana + Metarhizium anisopliae. The results show that the treatments with thiamethoxam + tefluthrin effectively reduced pest damage only at the critical stages of development. The current findings suggest that While some of these alternative methods offer good control, they may prove insufficient when applied individually. Hence, integrating them into a comprehensive pest management approach could be necessary for effectively safeguarding sugar beet yields. Further studies should explore potential additive or synergistic benefits to enhance these strategies. [ABSTRACT FROM AUTHOR]

Entomopathogenic fungi (EPF) represent the future of pest control, as they have the capacity to induce serious infections in insects, trigger epizootics, and effectively manage insect populations. The study aimed to identify and test indigenous EPF isolates against Tribolium confusum using soil samples from Northwestern and central Saudi Arabia. Galleria mellonella larvae were used as insect bait to collect EPF from soil samples from date palm groves, fruit groves, and maise fields. Only 27.3% of soil samples tested positive for palm grove-originating EPFs. Polymerase chain reaction (PCR) identified the cultured fungi as Metarhizium anisopliae and Beauveria bassiana. Tribolium confusum pathogenicity was assessed using nine local fungal isolates. Different fungus isolates were tested for their pathogenicity on larval and adult T. confusum. The results showed that M. anisopliae MaSA-2, MaSA-3, and MaSA-4 had infection rates of 85%, 85%, and 75%, respectively, in last-instar T. confusum larvae and 5%, 10%, and 20% in adults within seven days. On the other hand, B. bassiana BbSA-4 showed a 100% infection rate in T. confusum larvae, while BbSA-5, BbSA6, BbSA-7, BbSA-8, and BbSA-9 had infection rates of 70%, 80%, 80%, 75%, and 95%, respectively. In terms of adult mortality rates, B. bassiana BbSA-4, BbSA-5, BbSA-6, BbSA-7, BbSA-8, and BbSA-9 had rates of 25%, 35%, 35%, 20%, 20%, and 40%, respectively. The research findings show that indigenous B. bassiana and M. anisopliae fungus isolates successfully eliminated T. confusum larvae, although their effectiveness against adults was limited. [ABSTRACT FROM AUTHOR]

Insulin receptor substrate (IRS) proteins are key mediators in insulin signaling pathway. In social insect lives, IRS proteins played important roles in caste differentiation and foraging, but there function in disease defenses such as active immunization has not been reported yet. To investigate the issue, we successfully suppressed the IRS gene 3 days after dsRNA injection. Suppressing IRS gene increased the contents of glucose, trehalose, glycogen, and triglyceride and decreased the content of pyruvate in termites, and led to the metabolic disorder of glucose and lipids. IRS suppressing significantly enhanced grooming behaviors of nestmates of fungus-contaminated termites and hence increased the conidial load in the guts of the nestmates. Additionally, IRS suppressing led to significant downregulation of the immune genes Gram-negative bacteria-binding protein2 (GNBP2) and termicin and upregulation of the apoptotic gene caspase8 , and hence diminished antifungal activity of nestmates of fungus-contaminated termites. The above abnormal behavioral and physiological responses significantly decreased the survival rate of ds IRS -injected nestmates of the fungus-contaminated termites. These findings suggest that IRS is involved in regulation of active immunization in termites, providing a better understanding of the link between insulin signaling and the social immunity of termites. [ABSTRACT FROM AUTHOR]

BACKGROUND: Entomophagous fungi (EPF) not only directly kill insect pests, but also colonize plants and improve their resistance against pests. However, most previous research has focused on Beauveria bassiana and Metarhizium anisopliae, and there are few reports on whether other EPF can enhance resistance against pests via endogenous colonization. Herein, an EPF strain was isolated from diseased larvae of Spodoptera litura in a soybean field, and subjected to genome‐wide sequencing at the chromosomal level. The pathogenicity of the isolate toward various pest insects was evaluated, and the ability to colonize plants and induce resistance against phytopathogens and insect pests was tested. RESULTS: The purified isolate was identified as M. rileyi and designated MrS1Gz1–1. Biological assays revealed its strong pathogenicity toward five insect pests belonging to Lepidoptera and Hemiptera. Furthermore, the strain inhibited the growth of soil‐borne plant disease caused by Sclerotinia sclerotiorum in vitro. It colonized plants as an endophyte via soil application, thereby inducing plant resistance‐related genes against phytopathogen infection, and it disrupted the feeding selectivity of S. litura larvae. CONCLUSION: M. rileyi MrS1Gz1–1 has potential as a broad‐spectrum microbial control agent that can induce resistance against phytopathogens and insect pests feeding as an endotype. The complete genome provides a valuable resource for exploring host interactions. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Although most known viruses infecting fungi pathogenic to higher eukaryotes are asymptomatic or reduce the virulence of their host fungi, those that confer hypervirulence to entomopathogenic fungus still need to be explored. Here, we identified and studied a novel mycovirus in Metarhizium flavoviride, isolated from small brown planthopper (Laodelphax striatellus). Based on molecular analysis, we tentatively designated the mycovirus as Metarhizium flavoviride partitivirus 1 (MfPV1), a species in genus Gammapartitivirus, family Partitiviridae. MfPV1 has two double-stranded RNAs as its genome, 1,775 and 1,575 bp in size respectively, encapsidated in isometric particles. When we transfected commercial strains of Metarhizium anisopliae and Metarhizium pingshaense with MfPV1, conidiation was significantly enhanced (t test; P-value < 0. 01), and the significantly higher mortality rates of the larvae of diamondback moth (Plutella xylostella) and fall armyworm (Spodoptera frugiperda), two important lepidopteran pests were found in virus-transfected strains (ANOVA; P-value < 0.05). Transcriptomic analysis showed that transcript levels of pathogenesis-related genes in MfPV1-infected M. anisopliae were obviously altered, suggesting increased production of metarhizium adhesin-like protein, hydrolyzed protein, and destruxin synthetase. Further studies are required to elucidate the mechanism whereby MfPV1 enhances the expression ofpathogenesis-related genes and virulence ofMetarhizium to lepidopteran pests. This study presents experimental evidence that the transfection of other entomopathogenic fungal species with a mycovirus can confer significant hypervirulence and provides a good example that mycoviruses could be used as a synergistic agent to enhance the biocontrol activity of entomopathogenic fungi. [ABSTRACT FROM AUTHOR]

Background: Green bean, Phaseolus vulgaris L. (Fam.: Leguminosae) is a widely consumed grain legume prized for its edible seeds and pods. It is susceptible to infestations with various pests as insects and mites throughout the growing season. In this study, the efficacy of the predatory species, Chrysoperla carnea (Stephens), Phytoseiulus persimilis Athias-Henriot and the entomopathogenic fungus (EPF) Metarhizium anisopliae (Hypocreales: Clavicipitaceae), as well as conventional pesticides, Mospilan and Vertimec, were evaluated against the most important pests, mainly the whitefly, Bemisia tabaci (Genn.) and the two-spotted spider mite, Tetranychus urticae Koch, infesting the green beans cultivated at two locations Giza and El-Menoufia Governorates in Egypt. Results: The findings demonstrate that treatments using C. carnea and M. anisopliae effectively reduced the whitefly population, while pesticide treatments were comparatively less effective. In the Giza plots, at the end of the experiments, the use of both M. anisopliae and C. carnea showed high reductions in whitefly population (85.57 and 84.87%), respectively, while in El-Menoufia, C. carnea (97.74%) was the most effective treatment followed by M. anisopliae (90.32%). Pesticide treatment in this case yielded a reduction rate of (22.76 and 59.67%) in Giza and El-Menoufia plots, respectively. However, for spider mite control, P. persimilis proved to be the most effective treatment in Giza and El-Menoufia plots, reducing the spider mite population to 98.44 and 96.14%, respectively. Metarhizium anisopliae treatment also displayed moderate effectiveness, with reduction rates of 75.62 and 75.37% in Giza and El-Menoufia plots, respectively. In comparison, pesticide treatment showed low effectiveness, with reduction rates of only 23.92 and 53.16% in the two locations, respectively. Conclusion: Applications of the predator, C. carnea and the EPF, M. anisopliae were highly effective in reducing the population of whitefly, while the predator mite P. persimilis proved to be the most effective for controlling the spider mites. Overall, the study suggests that biocontrol agents, such as the predators and the EPF, can be considered as alternatives to synthetic chemical pesticides for controlling pests infesting green beans. [ABSTRACT FROM AUTHOR]

This study aimed to determine the effectiveness of three M. anisopliae isolates and tobacco extract with different densities and concentrations in controlling P. xylostella pest populations. This research was conducted from October to December 2022 at the Brenjonk Biological Agents Laboratory and the BPTPH Pasuruan Laboratory. This study employed a Randomized Block Design (RBD) with 16 treatments and 4 replications. Each treatment used 15 P. xylostella, which were tested for mortality, feeding inhibition, LC50, and LT50 with ANOVA and probit analysis of variance. The results showed that the highest mortality and inhibition were obtained from mushroom isolates from the Bojonegoro and Jatisari collections with a density of 108 and tobacco extract with a concentration of 50 ml/L. Meanwhile, the best LC50 value was found in the fungus isolated from the Jatisari collection, namely 4.99%, and the best LT50 was found in the mushroom isolate, the Bojonegoro collection, which was 133.09 hours. Based on the results of the study, it was shown that tobacco extract and the three isolates of the fungus M. anisopliae had the best ability to suppress P. xylostella pest populations. [ABSTRACT FROM AUTHOR]

The Guatemalan potato moth (Tecia solanivora) is one of the most destructive pests affecting potatoes in Colombia. Our objective was to evaluate the susceptibility of T. solanivora to a combination of entomopathogenic nematodes and entomopathogenic fungi, to study the interactions of these two agents under laboratory conditions. We evaluated the mortality of fourth instar larvae against 200 Infective Juveniles (IJs) per larva of ten strains of entomopathogenic nematodes, and 104–108 conidia per larva of Beauveria bassiana sensu lato Bb9205 and Metarhizium anisopliae sensu lato Ma9236. We found that Steinernema feltiae SC125 caused a mortality rate of 98.2% with an LC50 of 33 IJs/larva. M. anisopliae sensu lato Ma9236 achieved a mortality rate of 88.8% with an LC50 of 6.9 × 105 conidia/larva. We selected these strains for the interaction tests. We determined that the interaction test results were statistically additive regardless of any difference in application times of the entomopathogens and despite the observation of signs of infection in the hosts from both the nematode and the fungus. Despite not having found synergistic interactions between the evaluated control agents, these laboratory tests suggest a possible strategy of combining entomopathogenic nematode and fungi for biological management of the guatemalan moth T. solanivora in potato crops. [ABSTRACT FROM AUTHOR]

Four species of Trichoderma genius (T. harzianum, T. citrinoviride, T. viride and T. asperellum) and Metarhizium anisopliae were assessed for their effects on larval mortality and progeny production of Trogoderma granarium Everts (Coleoptera: Dermestidae). The fungal strains were tested at 2.0 × 106, 2.0 × 107, 2.0 × 108 and 2.0 × 109 spores/kg. The larval mortality of T. granarium was recorded after 7, 14 and 21 days of treatment. The emerged adults (F1) from treated larvae were counted after 30 days of treatment, while F2 and grain weight loss were examined after 80 days of treatment. In general, increasing of spore concentration of all fungal strains enhanced the larval mortality. After 21 of treatment, all fungal strains induced notable larval mortality particularly at the highest concentration (2.0 × 109 spores/kg) except T. viride (19.4%). The highest larval mortality was observed in treatments with M. anisopliae (82.1%) and T. citrinoviride (69.0%) at 2.0 × 109 spores/kg, respectively. Furthermore, all spore concentrations of tested fungi decreased the emerged adults (F1 and F2) with M. anisopliae, T. citrinoviride and T. harzianum being the most potent, particularly at 2.0 × 109 spores/kg. After 80 days, M. anisopliae and T. citrinoviride at 2.0 × 109 spores/kg induced the highest protection of wheat grains against larvae of T. granarium with weight loss of 0.7 and 1.3%, respectively. The outcomes of the present study indicate that the two fungi, M. anisopliae and T. citrinoviride are highly effective seed protectants against T. granarium and could be applied in the integrated pest management programs (IPM) of khapra beetle as effective biological control agents. [ABSTRACT FROM AUTHOR]

Simple Summary: Arthropods are a large and diverse group of animals, some of which are pests, while others transmit human and livestock diseases as they bite them to feed on blood. Chemical-based vector and pest control for decades has made many arthropods resistant to the chemicals used. So, there is a need to investigate other control agents, such as the biological, arthropod-killing fungus Metarhizium anisopliae, for the ability to infect and kill human and livestock disease-transmitting arthropods. In this study, we tested if the fungus M. anisopliae infects and kills mosquitoes, tsetse flies, and ticks. This was performed by finding out how long mosquitoes lived after being infected with fungus spores, as well as the amount of fungus spores suitable to lead to their death. The same optimal concentration of fungus that killed mosquitoes also successfully killed tsetse flies and ticks. We recommend that the fungus be formulated in a suitable concentration of 109 spores/mL and its ability to kill mosquitoes, tsetse flies, and ticks be confirmed in the field. This would lead to the development of a fungal formulation that can be sprayed on livestock such as cattle, for the exposure of multiple blood-feeding and disease-transmitting arthropods at the same time, to impact their survival. Arthropod vectors are responsible for a multitude of human and animal diseases affecting poor communities in sub-Saharan Africa. Their control still relies on chemical agents, despite growing evidence of insecticide resistance and environmental health concerns. Biorational agents, such as the entomopathogenic fungus Metarhizium anisopliae, might be an alternative for vector control. Recently, the M. anisopliae isolate ICIPE 7 has been developed into a commercial product in Kenya for control of ticks on cattle. We were interested in assessing the potential of controlling not only ticks but also disease-transmitting mosquitoes and tsetse flies using cattle as blood hosts, with the aim of developing a product for integrated vector management. Laboratory bioassays were carried out with M. anisopliae, isolate ICIPE 7 and isolate ICIPE 30, to compare efficacy against laboratory-reared Anopheles arabiensis. ICIPE 7 was further tested against wild Glossina fuscipes and Rhipicephalus spp. Dose–response tests were implemented, period of mosquito exposure was evaluated for effects on time to death, and the number of spores attached to exposed vectors was assessed. Exposure to 109 spores/mL of ICIPE 7 for 10 min resulted in a similar mortality of An. arabiensis as exposure to ICIPE 30, albeit at a slower rate (12 vs. 8 days). The same ICIPE 7 concentration also resulted in mortalities of tsetse flies (LT50: 16 days), tick nymphs (LT50: 11 days), and adult ticks (LT50: 20 days). Mosquito mortality was dose-dependent, with decreasing LT50 of 8 days at a concentration of 106 spores/mL to 6 days at 1010 spores/mL. Exposure period did not modulate the outcome, 1 min of exposure still resulted in mortality, and spore attachment to vectors was dose-dependent. The laboratory bioassays confirmed that ICIPE 7 has the potential to infect and cause mortality to the three exposed arthropods, though at slower rate, thus requiring further validation under field conditions. [ABSTRACT FROM AUTHOR]

The research aimed to isolate and characterize local entomopathogenic fungi from different agricultural ecosystems and assess their pathogenicity against cowpea aphid. Forty soil samples were collected from different ecosystems. In total, 38 fungal isolates were collected from the cadavers' exteriors of Galleria mellonella larvae. Microscopic observation of these fungal isolates revealed that they belonged to nine fungal genera among them Metarhizium sp. (3 isolates) and Beauveria sp. (5 isolates). Primary experiments showed different bioactivities of the isolates of the two selected genera against the target insect. Only Metarhizium isolates (M3 & M5) and Beauveria isolates (B1 & B5) were selected for further investigations. Identification of fungal cultures by morphological characters, molecular characterization, ITS sequencing, and NCBI Blast showed that two isolates were confirmed as M. anisopliae and two isolates as B. bassiana. The bioassay evaluation revealed that M. anisopliae (M3 and M5) and B. bassiana (B1) exhibited significant virulence against A. craccivora, as evidenced by their high LT50 and LC50 values and substantial sporulation on the cadavers. Therefore, they could be used for the formulation of pesticides targeting this pest on cowpea and other vegetable crops. [ABSTRACT FROM AUTHOR]

Nezara viridula (Linnaeus) (Hemiptera: Pentatomidae), commonly known as the southern green stink bug, poses a significant threat to agricultural productivity due to its destructive feeding nature which leads to substantial reductions in crop quality and yield. The entomopathogenic fungal genus Metarhizium (Hypocreales: Clavicipitaceae) has been known as a promising biological control agent with a wide range of arthropod hosts. Among them, Metarhizium anisopliae has shown potential as an alternative management strategy for controlling pests within crop ecosystems. This study aimed to assess the efficacy of different doses of M. anisopliae AUMC 3262 air-dried conidia (0.5, 2.5, and 5 g) against N. viridula adults and evaluate physiological and structural alternations under laboratory conditions. Results indicated that M. anisopliae AUMC 3262 dry conidia induced significant mortality in N. viridula in a time- and dose-dependent manner. Mortality rates reached 86.65%., 99.98% and 100 % of N. viridula after 5 and 6 days of exposure to 0.5, 2.5 and 5 g of dry conidia, respectively. Also, a significant reduction in the total soluble proteins was observed. Scanning Electron Microscope analysis confirmed deformation in the body structure of N. viridula, highlighting fungal penetration and proliferation within the insect's body. Overall, M. anisopliae AUMC 3262 is a virulent strain for N. viridula, which might qualify this strain to be used as a strong candidate for further investigation as a control agent for N. viridula. [ABSTRACT FROM AUTHOR]

The fall armyworm, Spodoptera frugiperda (J E Smith) is an emerging polyphagous pest. Its primary food is the corn cob, which suffers significant damage with its larvae skeletonizing or “windowing” the leaves. Only the larval stage is seriously risky and small and large communities as well as researchers have used a variety of control methods A number of low-cost mitigation strategies are available and these strategies include encouraging smallholder farmers to plant crops directly after major rainstorms, using cultural methods, such as intercropping and crop rotation, and avoiding some of the ineffective insecticides. Understanding the function of natural enemies in agricultural systems requires monitoring them in the field. Chelonus insularis, Pristomerus spinator, Cotesia marginiventris, Meteorus laphygmae, Chelonus sonorensis, Orius insidiosus, Chelonus texanus, Archytas marmoratus, Elenomus remus, and Trichogramma spp. are parasitoids of S. frugiperda, Metarhizium anisopliae, Erynia radicans, Fusarium solani, Nomuraea rileyi and Beauveria bassiana are the main entomopathogenic fungi. These biocontrol agents should be promoted to manage this pest in a sustainable manner. [ABSTRACT FROM AUTHOR]

The Asian long-horn beetle (ALB) is a serious wood-boring insect. Continuous isolation of different fungal strains is vital for using fungi for the control of ALB. The virulence and pathological characteristics of a new Metarhizium anisopliae strain DES3 isolated from the desert afforestation stands against the larvae of ALB were assessed in this study. The corrected mortality reached 100% at the conidial concentration of 109 and 108 conidia/mL, and 91.11 ± 4.44% at 107 conidia/mL. Similarly, the LC/LT showed high virulence as well. Meanwhile, the virulence of a commercial M. anisopliae strain against the ALB larvae was evaluated. The corrected mortality was only 33.33% at 109 conidia/mL, and less than 10% at 108 conidia/mL. The pathological characteristics after infection by the M. anisopliae strain DES3 were evident, mainly embodied in the rupture of the adipose tissue, muscle tissue, and midgut. But there was no obvious change after infection by the commercial M. anisopliae strain. In conclusion, these results establish that the M. anisopliae strain DES3 has high virulence in a dosage-dependent manner against ALB larvae, indicating the potential of fungal strain DES3 to be developed as biopesticide for biocontrol of A. glabripennis. [ABSTRACT FROM AUTHOR]

Metarhizium anisopliae is an important class of entomopathogenic fungi used for the biocontrol of insects, but its virulence is affected by insect immunity. We identified a novel FK506 binding protein gene that was differentially expressed between control and Metarhizium-treated Locusta migratoria manilensis. We hypothesized that this protein played an important role in Metarhizium infection of L. migratoria and could provide new insights for developing highly efficient entomopathogenic fungi. We, therefore, cloned the specific gene and obtained its purified protein. The gene was then named FKBP52 , and its dsRNA (dsFKBP52) was synthesized and used for gene interference. Bioassay results showed that the mortality of L. migratoria treated with dsFKBP52  + Metarhizium was significantly lower than that of other treatments. Furthermore, immune-related genes (MyD88 , Dorsal , Cactus , and Defensin) in L. migratoria treated with dsFKBP52  + Metarhizium showed significant upregulation compared to that treated with Metarhizium only. However, the activities of peroxidase (POD), superoxide dismutase (SOD), and calcineurin (CaN) showed fluctuations. These results suggest that the FKBP52 gene may play a crucial role in the innate immunity of L. migratoria. The effect of its silencing indicated that this immunity-related protein might be a potential target for insect biocontrol. [ABSTRACT FROM AUTHOR]

(1) Background: The mycophagous mite, Tyrophagus putrescentiae, was found to feed on entomopathogenic fungi (EPF) in our previous experiments, which seriously impacted the culture and preservation of fungal strains. Therefore, it is necessary to investigate the biological characteristics of the occurrence and damage to EPF. (2) Methods: The mite's growth and development and feeding preference were surveyed by comparative culture and observation; also, optical and electronic microscopies were employed. (3) Results: T. putrescentiae could survive normally after being fed on seven EPF species, including Purpureocillium lilacinum, Marquandii marquandii, Cordyceps fumosorosea, Beauveria bassiana, Metarhizium flavoviride, Lecanicillium dimorphum, and Metacordyceps chlamydosporia. The first four fungi were the mite's favorites with their greater feeding amount and shorter developmental duration. Interestingly, the mite could also feed on Metarhizium anisopliae and Metarhizium robertsii, but this led to the mite's death. After feeding on M. anisopliae and M. robertsii, the mites began to die after 24 h, and the mortality rate reached 100% by 72 h. Observation under optical microscopy and scanning electron microscopy revealed that the conidia of M. anisopliae and M. robertsii adhered to the mite's surface, but there was no evidence of penetration or invasion. However, dissection observation indicated that the two Metarhizium species germinate and grow within the mite's digestive tract, which implies that Metarhizium generalists with broad-spectrum hosts and the production of destruxins have acaricidal activity toward the mycophagous mites. [ABSTRACT FROM AUTHOR]

Fall armyworm (FAW), Spodoptera frugiperda is a generalist pest known to feed on more than 300 plant species, including major staple crops such as rice, maize and sorghum. Biological control of FAW using a combination of a major indigenous egg parasitoid Telenomus remus and entomopathogenic fungi was explored in this study. Metarhizium anisopliae strains (ICIPE 7, ICIPE 41, and ICIPE 78) and Beauveria bassiana ICIPE 621 which demonstrated effectiveness to combat the pest, were evaluated through direct and indirect fungal infection to assess their pathogenicity and virulence against T. remus adults, S. frugiperda eggs and their effects on T. remus parasitism rates. Metarhizium anisopliae ICIPE 7 and ICIPE 78 exhibited the highest virulence against T. remus adults with LT50 values >2 days. ICIPE 7 induced the highest T. remus mortality rate (81.40 ± 4.17%) following direct infection with dry conidia. Direct fungal infection also had a significant impact on parasitoid emergence, with the highest emergence rate recorded in the M. anisopliae ICIPE 7 treatment (42.50 ± 5.55%), compared to the control ± (83.25 ± 5.94%). In the indirect infection, the highest concentration of 1 x 109 conidia ml-1 of ICIPE 78 induced the highest mortality (100 ± 0.00%) of T. remus adults, and the highest mortality (51.25%) of FAW eggs, whereas the least FAW egg mortality (15.25%) was recorded in the lowest concentration 1 x 105 conidia ml-1 of ICIPE 41. The number of parasitoids that emerged and their sex ratios were not affected by the different fungal strain concentrations except in ICIPE 7 at high dose. This study showed that potential combination of both M. anisopliae and B. bassiana with T. remus parasitoid can effectively suppress FAW populations.