Metarhizium Brunneum Research Articles

Camponotusfloridanus Ants Incur a Trade-Off between Phenotypic Development and Pathogen Susceptibility from Their Mutualistic Endosymbiont Blochmannia.

Various insects engage in microbial mutualisms in which the reciprocal benefits exceed the costs. Ants of the genus Camponotus benefit from nutrient supplementation by their mutualistic endosymbiotic bacteria, Blochmannia, but suffer a cost in tolerating and regulating the symbiont. This cost suggests that the ants face secondary consequences such as susceptibility to pathogenic infection and transmission. In order to elucidate the symbiont’s effects on development and disease defence, Blochmannia floridanus was reduced in colonies of Camponotus floridanus using antibiotics. Colonies with reduced symbiont levels exhibited workers of smaller body size, smaller colony size, and a lower major-to-minor worker caste ratio, indicating the symbiont’s crucial role in development. Moreover, these ants had decreased cuticular melanisation, yet higher resistance to the entomopathogen Metarhizium brunneum, suggesting that the symbiont reduces the ants’ ability to fight infection, despite the availability of melanin to aid in mounting an immune response. While the benefits of improved growth and development likely drive the mutualism, the symbiont imposes a critical trade-off. The ants’ increased susceptibility to infection exacerbates the danger of pathogen transmission, a significant risk given ants’ social lifestyle. Thus, the results warrant research into potential adaptations of the ants and pathogens that remedy and exploit the described disease vulnerability.

Efficacy of entomopathogenic fungi against large pine weevil, Hylobius abietis, and their additive effects when combined with entomopathogenic nematodes

The large pine weevil Hylobius abietis is an important pest of reforestation in northern Europe. In field trials, we assessed the efficacy of entomopathogenic fungi (EPF) alone and in combination with entomopathogenic nematodes (EPN) against immature stages. We used commercial strains of Metarhizium brunneum and Beauveria bassiana and a strain of Beauveria caledonica isolated from the pest’s habitat. The EPN used were Steinernema carpocapsae and Heterorhabditis downesi. Efficacy was assessed by weevil infection in stumps and by numbers of adult weevils emerging in traps above the stumps. EPF infected up to 23% of pine weevils, at depths up to 25 cm below ground. Three EPF application methods were compared, but application method did not affect infection rates. When applied at half doses, EPF and EPN had additive effects: M. brunneum and S. carpocapsae caused a reduction in emergence of 58% relative to control, M. brunneum and H. downesi 93%, B. caledonica and H. downesi 86%. EPN alone offered good suppression of H. abietis populations and suppression by the mixture did not surpass suppression afforded by EPN alone. Our trials show B. caledonica is a promising species, rivalling the success of the other two species. Applied EPF, especially M. brunneum, can persist for at least 2 years in the forest setting. Additionally, different fungi can have differential action against weevils above versus below ground; therefore, EPF combinations may be beneficial. Based on our trials, further development of strains and application methods are required before EPF can be recommended for suppression of pine weevil.

Compatibility of foliage-dwelling predatory mites and mycoinsecticides, and their combined efficacy against western flower thrips Frankliniella occidentalis

Foliage-dwelling predatory mites and foliar applications of mycoinsecticides are commonly used in biological control programs for Western flower thrips. A laboratory study was designed to examine the compatibility of two foliage-dwelling predatory mites with two commercially available mycoinsecticides, followed by a greenhouse study to assess their combined efficacy against Western flower thrips, with a view to their concurrent use in an integrated strategy. The following commercially available biocontrol agents were evaluated: the predatory mites, Neoseiulus cucumeris (Oudemans) and Amblyseius swirskii (Athias-Henriot); and entomopathogenic fungi, Metarhizium anisopliae (Metschnikoff) Sorokin (now classified as Metarhizium brunneum) strain F52 and Beauveria bassiana (Balsamo) GHA strain. Mortality caused by the mycoinsecticides ranged from 0 to 15.98% in the laboratory studies. In the greenhouse, the relative efficacy of predatory mite slow-release breeding sachets, Met52 EC spray, and a combined application was determined. Under high pest pressure, Met52 EC-alone was not as effective as N. cucumeris-alone or the combination treatment over 8 weeks. Neoseiulus cucumeris-alone provided better control of thrips than Met52 EC, but in a mixed infestation of thrips and two-spotted spider mites, the combination treatment worked best overall; the spider mites were effectively suppressed by Met52 EC. Under low pest pressure in the experiment with A. swirskii, use of Met52 EC or A. swirskii sachets effectively suppressed thrips population growth; moreover, the combination treatment completely eliminated both thrips and spider mites.

Endophytic Metarhizium brunneum mitigates nutrient deficits in potato and improves plant productivity and vitality

Abstract There is growing evidence that entomopathogenic fungi play a role in plant growth promotion as colonizers of internal plant tissues. However, little is known about their potential to mitigate plant abiotic stress. Here, we investigated the influence of soil fertility on Metarhizium brunneum strain CB15 endophytism levels in potato plants linked to plant productivity and vitality. By application of encapsulated M. brunneum, the endophytism level, i.e. % presence of fungus in plant tissue, increased significantly under nutrient poor conditions. This correlated with significantly improved quantum yields of photosystem II. In addition, water use efficiency was increased with higher levels of M. brunneum endophytism. Furthermore, biomass, leaf area, nitrogen and phosphorus contents were enhanced indicating fungal nutrient mobilization and transfer. Our results provide first evidence for the role of entomopathogenic fungi in mitigating nutrient deficits in soil by improving plant productivity and vitality which may increase their use in plant protection strategies.

Presence of a generalist entomopathogenic fungus influences the oviposition behaviour of an aphid-specific predator

The predator Aphidoletes aphidimyza (Rondani) (Diptera: Cecidomyiidae) and the generalist entomopathogenic fungus Metarhizium brunneum Petch (Hypocreales: Clavicipitaceae) are effective biological control agents that can co-occur in pest management programmes. We exposed larvae to M. brunneum, on leaves and in soil, to explore possible outcomes of combining the two natural enemies. The number of emerging adults and their longevity were negatively affected by fungus in the soil. When exposed on leaves, adult emergence was unaffected but longevity was reduced. In choice experiments, females chose to lay eggs on fungus-free leaves: more control leaves had eggs and there were more eggs per leaf in the control. In no-choice experiments, the same frequency of females laid eggs in the treatment and control, but significantly more eggs were laid on the control leaves. Gravid A. aphidimyza can perceive M. brunneum and respond by choosing fungus-free oviposition sites, thus reducing contact between them.

Mortality of Solenopsis invicta Workers (Hymenoptera: Formicidae) After Indirect Exposure to Spores of Three Entomopathogenic Fungi

Mortality caused by indirect exposure to Metarhizium brunneum and Beauveria bassiana (GHA and NI8) to the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae), workers was evaluated. Groups of 50 workers were placed in one side of dual-box arenas. The opposite side of the arenas was lined with filter paper squares previously sprayed with unformulated purified spores (106 spores/ml) suspended in 0.2% Ethal TDA 3, HLB 8 of the three fungal strains, or untreated filter paper squares as the control. Daily observations were done for 1 wk to determine mortality. Dead ants from each treatment and control were collected, surface cleaned, and placed in PDA media and incubated at 27°C, 60% RH for 7 d to detect fungal growth. The presence of fungal growth in the dead ants confirmed that fungal spores infected workers while walking on the treated paper. In the M. brunneum and B. bassiana GHA treatments, 51.35 and 56.68% of the workers died, respectively, during days 1 and 2. However, only 9.47 and 35.96% of the mortality could be explained by fungal infection by M. brunneum and B. bassiana GHA, respectively. Most of the mortality observed in the B. bassiana NI8 treatment (84.48%) occurred later (between days 4–6) and most of this mortality occurring during day 4 (89.06%) could be explained by B. bassiana infection. Overall mortality was significantly higher in the B. bassiana NI8 treatment than the other two fungi tested and control. Potential application of these fungal strains for fire ant control are discussed.

Spore Acquisition and Survival of Ambrosia Beetles Associated with the Laurel Wilt Pathogen in Avocados after Exposure to Entomopathogenic Fungi.

Laurel wilt is a disease threatening the avocado industry in Florida. The causative agent of the disease is a fungus vectored by ambrosia beetles that bore into the trees. Until recently, management strategies for the vectors of the laurel wilt fungus relied solely on chemical control and sanitation practices. Beneficial entomopathogenic fungi (EPF) are the most common and prevalent natural enemies of pathogen vectors. Laboratory experiments demonstrated that commercial strains of EPF can increase the mortality of the primary vector, Xyleborus glabratus, and potential alternative vectors, Xylosandrus crassiusculus, Xyleborus volvulus and Xyleborus bispinatus (Coleoptera: Curculionidae: Scolytinae). Our study provides baseline data for three formulated commercially-available entomopathogenic fungi used as potential biocontrol agents against X. crassiusculus, X. volvulus and X. bispinatus. The specific objectives were to determine: (1) the mean number of viable spores acquired per beetle species adult after being exposed to formulated fungal products containing different strains of EPF (Isaria fumosorosea, Metarhizium brunneum and Beauveria bassiana); and (2) the median and mean survival times using paper disk bioassays. Prior to being used in experiments, all fungal suspensions were adjusted to 2.4 × 106 viable spores/mL. The number of spores acquired by X. crassiusculus was significantly higher after exposure to B. bassiana, compared to the other fungal treatments. For X. volvulus, the numbers of spores acquired per beetle were significantly different amongst the different fungal treatments, and the sequence of spore acquisition rates on X. volvulus from highest to lowest was I. fumosorosea > M. brunneum > B. bassiana. After X. bispinatus beetles were exposed to the different suspensions, the rates of acquisition of spores per beetle amongst the different fungal treatments were similar. Survival estimates (data pooled across two tests) indicated an impact for each entomopathogenic fungus per beetle species after exposure to a filter paper disk treated at the same fungal suspension concentration. Kaplan–Meier analysis (censored at day 7) revealed that each beetle species survived significantly shorter in bioassays containing disks treated with EPF compared to water only. This study demonstrated that ambrosia beetles associated with the laurel wilt pathogen in avocados are susceptible to infection by EPF under laboratory conditions. However, the EPF needs to be tested under field conditions to confirm their efficacy against the beetles.

Investigating the potential of an autodissemination system for managing populations of vine weevil, Otiorhynchus sulcatus (Coleoptera: Curculionidae) with entomopathogenic fungi.

Vine weevil, also known as black vine weevil, (Otiorhynchus sulcatus) is an economically important pest affecting soft fruit and nursery stock in temperate regions. We used laboratory and polytunnel experiments to investigate a novel control system based on autodissemination of spores of an entomopathogenic fungus to populations of adult vine weevils. The fungus was applied as a conidial powder, used on its own or formulated with talc, to a simple plastic refuge for vine weevils. The potential for adult weevils to disseminate the fungus was investigated first in polytunnel experiments using fluorescent powders applied to the refuge in lieu of fungal conidia. In this system, 88% of adult weevils came in contact with the powder within 48 h. When the powder was applied to five adult weevils that were then placed within a population of 35 potential recipients, it was transmitted on average to 75% of the recipient population within 7 days. Three isolates of entomopathogenic fungi (Beauveria bassiana isolate codes 433.99 and 1749.11 and Metarhizium brunneum isolate code 275.86), selected from a laboratory virulence screen. These three isolates were then investigated for efficacy when applied as conidial powders in artificial refuges placed among populations of adult weevils held in experimental boxes in the laboratory at 20 °C. Under this regime, the fungal isolates caused 70-90% mortality of adult weevils over 28 days. A final polytunnel experiment tested the efficacy of conidial powders of M. brunneum 275.86 placed in artificial refuges to increase vine weevil mortality. Overall weevil mortality was relatively low (26-41%) but was significantly higher in cages in which the conidial powders were placed in refuge traps than in cages with control traps. The lower weevil mortality recorded in the polytunnel experiment compared to the laboratory test was most likely a consequence of the greater amounts of inoculum required to kill adult weevils when conditions fluctuate between favourable and unfavourable temperatures e.g. below 15 °C. The potential of an autodissemination system for entomopathogenic fungi as a means of controlling vine weevil as part of an integrated pest management programme is discussed.

Entomopathogenic Nematodes and Fungi Virulence to Cowpea Curculio (Coleoptera: Curculionidae) Larvae1

Abstract Cowpea curculio, Chalcodermus aeneus (Boheman), is the major pest of southern peas or cowpea, Vigna unguiculata (L.). Throughout the southeastern United States, current management recommendations for cowpea curculio rely on insecticide applications; however, resistance has been observed. Entomopathogenic biocontrol agents, specifically nematodes in the genera Steinernema and Heterorhabditis and fungi in the genera Beauveria and Metarhizium, have shown efficacy with other curculio pests. We conducted laboratory bioassays to assess the virulence of entomopathogenic agents as a first step to provide alternative control strategies against cowpea curculio. Using controlled environmental conditions, we tested the effects of nematodes (Steinernema carpocapsae Weiser, Steinernema feltiae Filipjev, Heterorhabditis bacteriophora Poinar, and Heterorhabditis indica Poinar) and two fungi (Beauveria bassiana Balsamo and Metarhizium brunneum Petch) on survival and infectivity of cowpea curculio. Heterorhabditis...

Responses of entomopathogenic fungi to the mutagen 4-nitroquinoline 1-oxide

Survival of entomopathogenic fungi under solar ultraviolet (UV) radiation is paramount to the success of biological control of insect pests and disease vectors. The mutagenic compound 4-nitroquinoline 1-oxide (4-NQO) is often used to mimic the biological effects of UV radiation on organisms. Therefore, we asked whether tolerance to 4-NQO could predict tolerance to UV radiation in thirty isolates of entomopathogenic fungi and one isolate of a xerophilic fungus. A dendrogram obtained from cluster analyses based on the 50 and 90 % inhibitory concentrations (IC50 and IC90, respectively) divided the fungal isolates into six clusters numbered consecutively based on their tolerance to 4-NQO. Cluster 6 contained species with highest tolerance to 4-NQO (IC50 > 4.7 μM), including Mariannaea pruinosa, Lecanicillium aphanocladii, and Torrubiella homopterorum. Cluster 1 contained species least tolerant to 4-NQO (IC50 < 0.2 μM), such as Metarhizium acridum (ARSEF 324), Tolypocladium geodes, and Metarhizium brunneum (ARSEF 7711). With few exceptions, the majority of Metarhizium species showed moderate to low tolerances (IC50 between 0.4 and 0.9 μM) and were placed in cluster 2. Cluster 3 included species with moderate tolerance (IC50 between 1.0 and 1.2 μM). In cluster 4 were species with moderate to high tolerance (IC50 between 1.3 and 1.6 μM). Cluster 5 contained the species with high tolerance (IC50 between 1.9 and 4.0 μM). The most UV tolerant isolate of M. acridum, ARSEF 324, was the least tolerant to 4-NQO. Also, L. aphanocladii, which is very susceptible to UV radiation, showed high tolerance to 4-NQO. Our results indicate that tolerance to 4-NQO does not correlate with tolerance to UV radiation. Therefore this chemical compound is not a predictor of UV tolerance in entomopathogenic fungi.

Effects of endophytic entomopathogenic fungi on soybean aphid and identification of Metarhizium isolates from agricultural fields.

Terrestrial plants can harbor endophytic fungi that may induce changes in plant physiology that in turn affect interactions with herbivorous insects. We evaluated whether the application of entomopathogenic fungi Beauveria bassiana and Metarhizium brunneum to soybean seeds could become endophytic and affect interactions with soybean aphid (Aphis glycines Matsumura). It was found that A. glycines population sizes increased on plants with M. brunneum (strain F52) seed inoculum, but no significant effects were shown with analogous treatments with B. bassiana (strain GHA). Fungi recovered from soybean plant tissues indicate that endophytism was established, and that B. bassiana was more prevalent. Metarhizium brunneum was only recovered from stems, but B. bassiana was recovered from stems and leaves. This work confirms that some entomopathogenic fungi can be endophytic in soybean, however, some of these fungi may have a negative effect on the plants by increasing susceptibility of soybean to A. glycines. We also used DNA sequence data to identify species of Metarhizium obtained from agricultural fields in Iowa. Phylogenetic analyses, based on DNA sequence data, found that all isolates were Metarhizium robertsii, which is consistent with past studies indicating a cosmopolitan distribution and wide host range for this species. These results are important for understanding the dynamics of implementing environmentally sustainable measures for the control of pest insects.

Cellulase enhances endophytism of encapsulated Metarhizium brunneum in potato plants

The recent discovery that entomopathogenic fungi can grow endophytically in plant tissues has spurred research into novel plant protection measures. However, current applications of fungi aiming at endophytism mostly lack targeted formulation strategies resulting in low efficacy. Here, we aimed at enhancing Metarhizium brunneum CB15 endophytism in potato plants by (i) improvement of fungal growth from beads and (ii) cellulase formation or addition to encapsulated mycelium. We found that beads supplemented with cellulose alone or in addition with inactivated baker's yeast exhibited cellulase activity and increased mycelial growth by 12.6 % and 13.6 %, respectively. Higher enzymatic activity achieved by cellulase co-encapsulation promoted a shift from mycelial growth to spore formation with maximum numbers of 2.5 × 108 ± 6.1 × 107 per bead. This correlated with improved endophytism in potato plants by 61.2 % compared to non-supplemented beads. Our study provides first evidence that customized formulations of fungal entomopathogens with enzymes can improve endophytism and this may increase efficacy in plant protection strategies against herbivorous pests.

Improving biocontrol of black vine weevil (Otiorhynchus sulcatus) with entomopathogenic fungi in growing media by incorporating spent mushroom compost

Amending a peat-based growing medium with 10% v/v spent mushroom compost, a source of fungal chitin and other nutrients, prolonged the persistence of entomopathogenic fungi (Metarhizium brunneum Petsch and Beauveria bassiana (Balsamo) Vuillemin; Hypocreales: Clavicipitaceae). This resulted in improved efficacy of M. brunneum against black vine weevil, Otiorhynchus sulcatus F. (Coleoptera: Curculionidae) larvae compared with using inoculum without spent mushroom compost. B. bassiana only controlled larvae when used in combination with spent mushroom compost (75 ± 7% reduction in live larvae). Mixing entomopathogenic fungal inoculum with spent mushroom compost and growing medium was as effective in controlling black vine weevil larvae as using spent mushroom compost colonised with M. brunneum or B. bassiana in the growing medium (80 ± 12% reduction in live larvae). The former method is preferable since it does not require production and storage of colonised spent mushroom compost, or registration of new substrate formulations of M. brunneum or B. bassiana.

Significant mortality of eggs and young larvae of two pine processionary moth species due to the entomopathogenic fungus Metarhizium brunneum

ABSTRACTBioassays were conducted to determine the susceptibility of egg masses and young larvae of two pine processionary moth species, Thaumetopoea pityocampa and Thaumetopoea wilkinsoni, to two strains (ARSEF4556, V275) of the entomopathogenic fungus Metarhizium brunneum. Mortality of treated eggs by both strains ranged from 96% to 99% but not all of this was caused by M. brunneum since control groups also experienced egg mortality due to saprophytic fungi. Still, larvae hatched in the laboratory from eggs treated with M. brunneum were all killed by this fungus, acquiring M. brunneum conidia, whereas larval mortality was 0% in the control groups. Young larvae of both pine processionary moth species were also highly susceptible to ARSEF4556 and V275 with larval mortality ranging between 94% and 100%, 8 days post-inoculation, with the vast majority of larvae being killed within the first 2–4 days. Larval mortality was dose dependent. Results were consistent across the two pine processionary moth species, showing that the pathogenicity of M. brunneum to both eggs and young larvae might be promising for biological control of these insect pests. The study also showed that non-target parasitoids of pine processionary moth eggs were also susceptible to M. brunneum. Further work is required to understand and reduce the M. brunneum effect on non-target insects.

Combined use of the entomopathogenic fungus, Metarhizium brunneum, and the mosquito predator, Toxorhynchites brevipalpis, for control of mosquito larvae: Is this a risky biocontrol strategy?

Mosquitoes transmit several diseases, which are of global significance (malaria, dengue, yellow fever, Zika). The geographic range of mosquitoes is increasing due to climate change, tourism and trade. Both conidial and blastospore formulations of the entomopathogenic fungus, Metarhizium brunneum ARSEF 4556, are being investigated as mosquito larvicides. However, concerns have been raised over possible non-target impacts to arthropod mosquito predators such as larvae of Toxorhynchites brevipalpis which feed on larvae of mosquito vector species. Laboratory-based, small container bioassays showed, that T. bevipalpis larvae are susceptible to relatively high concentrations (i.e. ≥107 spores ml-1) of inoculum with blastospores being significantly more virulent than conidia. At lower concentrations (e.g. <107 spores ml-1), it appears that M. brunneum complements T. brevipalpis resulting in higher control than if either agent was used alone. At a concentration of 105 spores ml-1, the LT50 of for conidia and blastospores alone was 5.64 days (95% CI: 4.79-6.49 days) and 3.89 days (95% CI: 3.53-4.25 days), respectively. In combination with T. brevipalpis, this was reduced to 3.15 days (95% CI: 2.82-3.48 days) and 2.82 days (95% CI: 2.55-3.08 days). Here, combined treatment with the fungus and predator was beneficial but weaker than additive. At 107 and 108 blastospores ml-1, mosquito larval mortality was mostly due to the fungal pathogen when the predator was combined with blastospores. However, with conidia, the effects of combined treatment were additive/synergistic at these high concentrations. Optimisation of fungal concentration and formulation will reduce: (1) risk to the predator and (2) application rates and costs of M. brunneum for control of mosquito larvae.

The Xenon Test Chamber Q-SUN® for testing realistic tolerances of fungi exposed to simulated full spectrum solar radiation

The low survival of insect-pathogenic fungi when used for insect control in agriculture is mainly due to the deleterious effects of ultraviolet radiation and heat from solar irradiation. In this study, conidia of 15 species of entomopathogenic fungi were exposed to simulated full-spectrum solar radiation emitted by a Xenon Test Chamber Q-SUN XE-3-HC 340S (Q-LAB® Corporation, Westlake, OH, USA), which very closely simulates full-spectrum solar radiation. A dendrogram obtained from cluster analyses, based on lethal time 50 % and 90 % calculated by Probit analyses, separated the fungi into three clusters: cluster 3 contains species with highest tolerance to simulated full-spectrum solar radiation, included Metarhizium acridum, Cladosporium herbarum, and Trichothecium roseum with LT50 > 200 min irradiation. Cluster 2 contains eight species with moderate UV tolerance: Aschersonia aleyrodis, Isaria fumosorosea, Mariannaea pruinosa, Metarhizium anisopliae, Metarhizium brunneum, Metarhizium robertsii, Simplicillium lanosoniveum, and Torrubiella homopterorum with LT50 between 120 and 150 min irradiation. The four species in cluster 1 had the lowest UV tolerance: Lecanicillium aphanocladii, Beauveria bassiana, Tolypocladium cylindrosporum, and Tolypocladium inflatum with LT50 < 120 min irradiation. The QSUN Xenon Test Chamber XE3 is often used by the pharmaceutical and automotive industry to test light stability and weathering, respectively, but it was never used to evaluate fungal tolerance to full-spectrum solar radiation before. We conclude that the equipment provided an excellent tool for testing realistic tolerances of fungi to full-spectrum solar radiation of microbial agents for insect biological control in agriculture.

Metarhizium brunneum (Ascomycota; Hypocreales) Treatments Targeting Olive Fly in the Soil for Sustainable Crop Production.

Soil treatments with Metarhizium brunneum EAMa 01/58-Su strain conducted in both Northern and Southern Spain reduced the olive fly (Bactrocera oleae) population density emerging from the soil during spring up to 70% in treated plots compared with controls. A model to determine the influence of rainfall on the conidial wash into different soil types was developed, with most of the conidia retained at the first 5 cm, regardless of soil type, with relative percentages of conidia recovered ranging between 56 and 95%. Furthermore, the possible effect of UV-B exposure time on the pathogenicity of this strain against B. oleae adults coming from surviving preimaginals and carrying conidia from the soil at adult emergence was also evaluated. The UV-B irradiance has no significant effect on M. brunneum EAMa 01/58-Su pathogenicity with B. oleae adult mortalities of 93, 90, 79, and 77% after 0, 2, 4, and 6 of UV-B irradiance exposure, respectively. In a next step for the use of these M. brunneum EAMa 01/58-Sun soil treatments within a B. oleae IPM strategy, its possible effect of on the B. oleae cosmopolitan parasitoid Psyttalia concolor, its compatibility with the herbicide oxyfluorfen 24% commonly used in olive orchards and the possible presence of the fungus in the olive oil resulting from olives previously placed in contact with the fungus were investigated. Only the highest conidial concentration (1 × 108 conidia ml−) caused significant P. concolor adult mortality (22%) with enduing mycosis in 13% of the cadavers. There were no fungal propagules in olive oil samples resulting from olives previously contaminated by EAMa 01/58-Su conidia. Finally, the strain was demonstrated to be compatible with herbicide since the soil application of the fungus reduced the B. oleae population density up to 50% even when it was mixed with the herbicide in the same tank. The fungal inoculum reached basal levels 4 months after treatments (1.6 × 103 conidia g soil−1). These results reveal both the efficacy and environmental and food safety of this B. oleae control method, protecting olive groves and improving olive oil quality without negative effects on the natural enemy P. concolor.

Destructive disinfection of infected brood prevents systemic disease spread in ant colonies.

In social groups, infections have the potential to spread rapidly and cause disease outbreaks. Here, we show that in a social insect, the ant Lasius neglectus, the negative consequences of fungal infections (Metarhizium brunneum) can be mitigated by employing an efficient multicomponent behaviour, termed destructive disinfection, which prevents further spread of the disease through the colony. Ants specifically target infected pupae during the pathogen's non-contagious incubation period, utilising chemical 'sickness cues' emitted by pupae. They then remove the pupal cocoon, perforate its cuticle and administer antimicrobial poison, which enters the body and prevents pathogen replication from the inside out. Like the immune system of a metazoan body that specifically targets and eliminates infected cells, ants destroy infected brood to stop the pathogen completing its lifecycle, thus protecting the rest of the colony. Hence, in an analogous fashion, the same principles of disease defence apply at different levels of biological organisation.

Species of the Metarhizium anisopliae complex with diverse ecological niches display different susceptibilities to antifungal agents

Species of the Metarhizium anisopliae complex are globally ubiquitous soil-inhabiting and predominantly insect-pathogenic fungi. The Metarhizium genus contains species ranging from specialists, such as Metarhizium acridum that only infects acridids, to generalists, such as M. anisopliae, Metarhizium brunneum, and Metarhizium robertsii that infect a broad range of insects and can also colonize plant roots. There is little information available about the susceptibility of Metarhizium species to clinical and non-clinical antifungal agents. We determined the susceptibility of 16 isolates comprising four Metarhizium species with different ecological niches to seven clinical (amphotericin B, ciclopirox olamine, fluconazole, griseofulvin, itraconazole, tebinafine, and voriconazole) and one non-clinical (benomyl) antifungal agents. All isolates of the specialist M. acridum were clearly more susceptible to most antifungals than the isolates of the generalists M. anisopliae sensu lato, M. brunneum, and M. robertsii. All isolates of M. anisopliae, M. brunneum, and M. robertsii were resistant to fluconazole and some were also resistant to amphotericin B. The marked differences in susceptibility between the specialist M. acridum and the generalist Metarhizium species suggest that this characteristic is associated with their different ecological niches, and may assist in devising rational antifungal treatments for the rare cases of mycoses caused by Metarhizium species.

Entomopathogens in conjunction with imidacloprid could be used to manage wireworms (Coleoptera: Elateridae) on spring wheat

AbstractWe examined the effect of biopesticides used alone, mixed with other biopesticides, or in conjunction with an imidacloprid against wireworms (Coleoptera: Elateridae) in spring wheat Triticum aestivum Linnaeus (Poaceae) (variety: Duclair). The study was conducted at Ledger and Valier, Montana, United States of America in 2015 and 2016. Ten biopesticides (spinosad, azadirachtin, pyrethrin, Beauveria bassiana (Balsamo-Crivelli) Vuillemin (Fungi: Clavicipitaceae) GHA (Mycotrol), B. bassiana ANT-03 (BioCeres), Chromobacterium subtsugae Martin et al. (Bacteria: Neisseriaceae), Burkholderia Yabuuchi et al. (Burkholderiaceae) species, Metarhizium brunneum Petch (Fungi: Clavicipitaceae) ESC1 (MbESC1), and M. brunneum F52 (MetF52) as microsclerotial and corn grit-based granules) were tested in addition to thimet and imidacloprid. Treatment efficacy was based on plant stand protection, wireworm populations, and yield. In 2015, there was considerable variation between sites in treatment efficacy. Mycotrol, BioCeres, MetF52+spinosad, and MetF52+imidacloprid applications protected seedlings from wireworm damage better than the control at Ledger, while only MetF52 at Valier. Wireworm populations were significantly higher with Mycotrol, spinosad, MetF52+spinosad, MbESC1 (25 g/L), and MetF52 treatments, compared with control, at 14 and 28 days post application at Ledger, but without effect at Valier, 2015. We found significantly higher yield in plots treated with imidacloprid+MetF52 and Mycotrol+azadirachtin (Xpulse) compared with control at Ledger. In 2016, no significant treatment effects were observed at either site. In summary, this study provides insight on what treatments should be explored in more detail despite variable results.