Study on the virulence of Metarhizium anisopliae against Spodoptera frugiperda (J. E. Smith, 1797).

This study examined the impact of Metarhizium anisopliae (Hypocreales: Clavicipitaceae) conidia on the eggs, larvae, pupae, and adults of Spodoptera frugiperda. The results showed that eggs, larvae, pupae, and adults exhibited mortality rates that were dependent on the dose. An increased amount of conidia (1.5 × 109 conidia/mL) was found to be toxic to larvae, pupae, and adults after 9 days of treatment, resulting in a 100% mortality rate in eggs, 98% in larvae, 76% in pupae, and 85% in adults. A study using earthworms as bioindicators found that after 3 days of exposure, M. anisopliae conidia did not cause any harmful effects on the earthworms. In contrast, the chemical treatment (positive control) resulted in 100% mortality at a concentration of 40 ppm. Histopathological studies showed that earthworm gut tissues treated with fungal conidia did not show significant differences compared with those of the negative control. The gut tissues of earthworms treated with monocrotophos exhibited significant damage, and notable differences were observed in the chemical treatment. The treatments with 70 and 100 µg/mL solutions of Eudrilus eugeniae epidermal mucus showed no fungal growth. An analysis of the enzymes at a biochemical level revealed a decrease in the levels of acetylcholinesterase, α-carboxylesterase, and ß-carboxylesterase in S. frugiperda larvae after exposure to fungal conidia. This study found that M. anisopliae is effective against S. frugiperda, highlighting the potential of this entomopathogenic fungus in controlling this agricultural insect pest.

No evidence for competition over floral resources between winter-active parasitoids and pollinators in agroecosystems.

Warming temperate winters are resulting in increased insect winter activity. With modern agroecosystems largely homogenous, characterised by low floral diversity, competitive interactions may arise between flower-visiting species, with potential implications for the ecosystem services they provide (e.g. biological control and pollination). Flower strips may be implemented during winter months to support flower-visiting insects and enhance ecosystem service provision. Employing field trials conducted in Brittany, France between 2019 and 2021 and laboratory cage experiments, the current study examined the impact of winter flower strips on aphid biological control performed by parasitoid wasps and the potential for competitive interactions between winter-active parasitoids and pollinators. Results revealed that parasitism rate was not enhanced by the presence of winter flower strips. This lack of effect was not the consequence of pollinator presence, and the current study found no effect of pollinator abundance on parasitism rate. Flower strips may thus be implemented during winter months to support nectar-feeding insects when floral resources are scarce, with no evidence of exploitative competition between pollinators and parasitoids, nor a detrimental impact on biological control provision.

Biocontrol effect of entomopathogenic fungi Metarhizium anisopliae ethyl acetate-derived chemical molecules: An eco-friendly anti-malarial drug and insecticide.

Insect pests represent a major threat to human health and agricultural production. With a current over-dependence on chemical insecticides in the control of insect pests, leading to increased chemical resistance in target organisms, as well as side effects on nontarget organisms, the wider environment, and human health, finding alternative solutions is paramount. The employment of entomopathogenic fungi is one such potential avenue in the pursuit of greener, more target-specific methods of insect pest control. To this end, the present study tested the chemical constituents of Metarhizium anisopliae fungi against the unicellular protozoan malaria parasite Plasmodium falciparum, the insect pests Anopheles stephensi Listen, Spodoptera litura Fabricius, and Tenebrio molitor Linnaeus, as well as the nontarget bioindicator species, Eudrilus eugeniae Kinberg. Fungal crude chemical molecules caused a noticeable anti-plasmodial effect against P. falciparum, with IC50 and IC90 values of 11.53 and 7.65 µg/mL, respectively. The crude chemical molecules caused significant larvicidal activity against insect pests, with LC50 and LC90 values of 49.228-71.846 µg/mL in A. stephensi, 32.542-76.510 µg/mL in S. litura, and 38.503-88.826 µg/mL in T. molitor at 24 h posttreatment. Based on the results of the nontarget bioassay, it was revealed that the fungal-derived crude extract exhibited no histopathological sublethal effects on the earthworm E. eugeniae. LC-MS analysis of M. anisopliae-derived crude metabolites revealed the presence of 10 chemical constituents. Of these chemicals, three major chemical constituents, namely, camphor (15.91%), caprolactam (13.27%), and monobutyl phthalate (19.65%), were highlighted for potential insecticidal and anti-malarial activity. The entomopathogenic fungal-derived crude extracts thus represent promising tools in the control of insect pests and malarial parasites.

First report on the enzymatic and immune response of Metarhizium majus bag formulated conidia against Spodoptera frugiperda: An ecofriendly microbial insecticide.

Entomopathogenic fungi from microbial sources are a powerful tool for combating insecticide resistance in insect pests. The purpose of the current study was to isolate, identify, and evaluate bag-formulated entomopathogenic fungal conidial virulence against insect pests. We further investigated the enzymatic responses induced by the entomopathogenic fungi as well as the effect on a non-target species. Entomopathogenic fungi were isolated from the Palamalai Hills, India, using the insect bait method, and the Metarhizium majus (MK418990.1) entomopathogen was identified using biotechnological techniques (genomic DNA isolation and 18S rDNA amplification). Bag-formulated fungal conidial efficacy (2.5 × 103, 2.5 × 104, 2.5 × 105, 2.5 × 106, and 2.5 × 107 conidia/ml) was evaluated against third instar larvae of Spodoptera frugiperda at 3, 6, 9, and 12 days of treatment, and acid and alkaline phosphatases, catalase, and superoxide dismutase enzymatic responses were evaluated at 3 days post-treatment. After 12 days of treatment, non-target assays on the earthworm Eudrilus eugeniae were performed using an artificial soil assay. Results of the bag formulated fungal conidial treatment showed that S. frugiperda had high susceptibility rates at higher concentrations (2.5 × 107 conidia/ml) of M. majus. Lower concentration of 2.5 × 103 conidia/ml caused 68.6% mortality, while 2.5 × 107 conidia/ml caused 100% mortality at 9 days post treatment. Investigation into enzymatic responses revealed that at 3 days post M. majus conidia exposure (2.5 × 103 conidia/ml), insect enzyme levels had significantly changed, with acid and alkaline phosphatases, and catalase enzymes significantly reduced and superoxide dismutase enzymes significantly raised relative to the control. After 12 days of treatment, no sublethal effects of M. majus conidia were observed on E. eugeniae, with no observed damage to gut tissues including lumen and epithelial cells, the nucleus, setae, coelom, mitochondria, and muscles. This study offers support for the use of fungal conidia in the target-specific control of insect pests.

Landscape effects on the thermotolerance of carabid beetles and the role of behavioral thermoregulation.

Physiological thermotolerance and behavioral thermoregulation are central to seasonal cold adaptation in ectothermic organisms. For species with enhanced mobility, behavioral responses may be of greater importance in the cold stress response. Employing the carabid beetles as a study organism, the current study compared physiological thermotolerance and behavioral thermoregulation in carabid species inhabiting cereal fields in different landscape contexts, from fine grain heterogeneous "complex" landscapes to homogenous "simple" landscapes. Physiological thermotolerance was determined via measurement of the CTmin and chill coma temperature. Behavioral responses to cold temperature exposure were determined employing a purpose built arena, and thoracic temperature measured to estimate the efficacy of the behavior as a form of behavioral thermoregulation. Results revealed an influence of landscape composition on the cold tolerance of carabid beetles, although species differed in their sensitivity to landscape intensification. A reduced effect of landscape on the thermotolerance of larger carabid beetles was observed, thought to be the consequence of greater mobility preventing local acclimation to microclimatic variation along the landscape intensification gradient. Investigation into behavioral thermoregulation of the 3 largest species revealed burrowing behavior to be the main behavioral response to cold stress, acting to significantly raise carabid body temperature. This finding highlights the importance of behavioral thermoregulation as a strategy to evade cold stress. The use of behavioral thermoregulation may negate the need to invest in physiological thermotolerance, further offering explanation for the lack of landscape effect on the physiological thermotolerance of larger carabids.

Toxicity of Metarhizium flavoviride conidia virulence against Spodoptera litura (Lepidoptera: Noctuidae) and its impact on physiological and biochemical activities.

Insect pests of agricultural crops have establish immunological tolerance against fungal infection caused by pathogens via different humoral and cellular processes. Fungal infection can be prevented by insect antioxidant and detoxifying enzymes, but there is no clear understanding of how they physiologically and biochemically interact. Our study aims to examine the antioxidant and detoxifying enzyme defense systems of the pest insect Spodoptera litura in response to infection by Metarhizium flavoviride. At 48 h following exposure to M. flavoviride, antioxidant enzyme levels were modified, and phenoloxidase and total hemocyte count were decreased significantly. The amount of detoxifying enzymes increased significantly. M. flavoviride appears to directly affect the S. litura immune system and results in decreased immunity. In a bioassay, M. flavoviride was found to be harmful to S. litura larvae in their third and fourth instar stage. M. flavoviride may be an effective tool in the control of S. litura larvae. Such entomopathogenic fungi represent cheaper, pollution free, target specific, promising alternatives to synthetic chemical tools in the for control insect pests.

Measuring the evolutionary potential of a winter-active parasitic wasp to climate change.

In temperate climates, as a consequence of warming winters, an increasing number of ectothermic species are remaining active throughout winter months instead of diapausing, rendering them increasingly vulnerable to unpredictable cold events. One species displaying a shift in overwintering strategy is the parasitoid wasp and biological control agent Aphidius avenae. The current study aimed to better understand the consequence of a changing overwintering strategy on the evolutionary potential of an insect population to adapt to the cold stress events, set to increase in frequency, even during milder winters. Using a parental half-sibling breeding design, narrow-sense heritability of the cold tolerance, morphology and longevity of A. avenae was estimated. The heritability of cold tolerance was estimated at 0.07 (CI95% = [0.00; 0.25]) for the Critical Thermal Minima (CTmin) and 0.11 (CI95% = [0.00; 0.34]) for chill coma temperature; estimates much lower than those obtained for morphological traits (tibia length 0.20 (CI95% = [0.03; 0.37]); head width 0.23 (CI95% = [0.09; 0.39]); wing surface area 0.28 (CI95% = [0.11; 0.47])), although comparable with the heritability estimate of 0.12 obtained for longevity (CI95% = [0.00; 0.25]). The heritability estimates obtained thus suggest that A. avenae possesses low adaptive potential against cold stress. If such estimates are indicative of the evolutionary potential of A. avenae cold tolerance, more emphasis may be placed on adaptive phenotypic plasticity at the individual level to persist in a changing climate, with potential implications for the biological control function they provide.

Assessment of neuropeptide binding sites and the impact of biostable kinin and CAP2b analogue treatment on aphid (Myzus persicae and Macrosiphum rosae) stress tolerance.

BACKGROUND: Neuropeptides are regulators of critical life processes in insects and, due to their high specificity, represent potential targets in the development of greener insecticidal agents. Fundamental to this drive is understanding neuroendocrine pathways that control key physiological processes in pest insects and the screening of potential analogues. The current study investigated neuropeptide binding sites of kinin and CAPA (CAPA-1) in the aphids Myzus persicae and Macrosiphum rosae and the effect of biostable analogues on aphid fitness under conditions of desiccation, starvation and thermal (cold) stress. RESULTS: M. persicae and M. rosae displayed identical patterns of neuropeptide receptor mapping along the gut, with the gut musculature representing the main target for kinin and CAPA-1 action. While kinin receptor binding was observed in the brain and VNC of M. persicae, this was not observed in M. rosae. Furthermore, no CAPA-1 receptor binding was observed in the brain and VNC of either species. CAP2b/PK analogues (with CAPA receptor cross-activity) were most effective in reducing aphid fitness under conditions of desiccation and starvation stress, particularly analogues 1895 (2Abf-Suc-FGPRLa) and 2129 (2Abf-Suc-ATPRIa), which expedited aphid mortality. All analogues, with the exception of 2139-Ac, were efficient at reducing aphid survival under cold stress, although were equivalent in the strength of their effect. CONCLUSION: In demonstrating the effects of analogues belonging to the CAP2b neuropeptide family and key analogue structures that reduce aphid fitness under stress conditions, this research will feed into the development of second generation analogues and ultimately the development of neuropeptidomimetic-based insecticidal agents. © 2019 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Renal neuroendocrine control of desiccation and cold tolerance by Drosophila suzukii.

BACKGROUND: Neuropeptides are central to the regulation of physiological and behavioural processes in insects, directly impacting cold and desiccation survival. However, little is known about the control mechanisms governing these responses in Drosophila suzukii. The close phylogenetic relationship of D. suzukii with Drosophila melanogaster allows, through genomic and functional studies, an insight into the mechanisms directing stress tolerance in D. suzukii. RESULTS: Capability (Capa), leucokinin (LK), diuretic hormone 44 (DH44 ) and DH31 neuropeptides demonstrated a high level of conservation between D. suzukii and D. melanogaster with respect to peptide sequences, neuronal expression, receptor localisation, and diuretic function in the Malpighian tubules. Despite D. suzukii's ability to populate cold environments, it proved sensitive to both cold and desiccation. Furthermore, in D. suzukii, Capa acts as a desiccation- and cold stress-responsive gene, while DH44 gene expression is increased only after desiccation exposure, and the LK gene after nonlethal cold stress recovery. CONCLUSION: This study provides a comparative investigation into stress tolerance mediation by neuroendocrine signalling in two Drosophila species, providing evidence that similar signalling pathways control fluid secretion in the Malpighian tubules. Identifying processes governing specific environmental stresses affecting D. suzukii could lead to the development of targeted integrated management strategies to control insect pest populations. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The effect of landscape complexity and microclimate on the thermal tolerance of a pest insect.

Landscape changes are known to exacerbate the impacts of climate change. As such, understanding the combined effect of climate and landscape on agroecosystems is vital if we are to maintain the function of agroecosystems. This study aimed to elucidate the effects of agricultural landscape complexity on the microclimate and thermal tolerance of an aphid pest to better understand how landscape and climate may interact to affect the thermal tolerance of pest species within the context of global climate change. Meteorological data were measured at the landscape level, and cereal aphids (Sitobion avenae, Metopolophium dirhodum and Rhopalosiphum padi) sampled, from contrasting landscapes (simple and complex) in winter 2013/2014 and spring 2014 in cereal fields of Brittany, France. Aphids were returned to the laboratory and the effect of landscape of origin on aphid cold tolerance (as determined by CTmin ) was investigated. Results revealed that local landscape complexity significantly affected microclimate, with simple homogenous landscapes being on average warmer, but with greater temperature variation. Landscape complexity was shown to impact aphid cold tolerance, with aphids from complex landscapes being more cold tolerant than those from simple landscapes in both winter and spring, but with differences among species. This study highlights that future changes to land use could have implications for the thermal tolerance and adaptability of insects. Furthermore, not all insect species respond in a similar way to microhabitat and microclimate, which could disrupt important predator-prey relationships and the ecosystem service they provide.

Why is there no impact of the host species on the cold tolerance of a generalist parasitoid?

For generalist parasitoids such as those belonging to the Genus Aphidius, the choice of host species can have profound implications for the emerging parasitoid. Host species is known to affect a variety of life history traits. However, the impact of the host on thermal tolerance has never been studied. Physiological thermal tolerance, enabling survival at unfavourable temperatures, is not a fixed trait and may be influenced by a number of external factors including characteristics of the stress, of the individual exposed to the stress, and of the biological and physical environment. As such, the choice of host species is likely to also have implications for the thermal tolerance of the emerging parasitoid. The current study aimed to investigate the effect of cereal aphid host species (Sitobion avenae, Rhopalosiphum padi and Metopolophium dirhodum) on adult thermal tolerance, in addition to sex and size, of the aphid parasitoids Aphidius avenae, Aphidius matricariae and Aphidius rhopalosiphi. Results revealed no effect of host species on the cold tolerance of the emerging parasitoid, as determined by CTmin and Chill Coma, for all parasitoid species. Host species significantly affected the size of the emerging parasitoid for A. rhopalosiphi only, with individuals emerging from R. padi being significantly larger than those emerging from S. avenae, although this did not correspond to a difference in thermal tolerance. Furthermore, a significant difference in the size of male and female parasitoids was observed for A. avenae and A. matricariae, although, once again this did not correspond to a difference in cold tolerance. It is suggested that potential behavioural thermoregulation via host manipulation may act to influence the thermal environment experienced by the wasp and thus wasp thermal tolerance and, in doing so, may negate physiological thermal tolerance or any impact of the aphid host.

Potential Host Manipulation by the Aphid Parasitoid Aphidius avenae to Enhance Cold Tolerance.

During parasitoid development, the immature parasitoid is confined to the host species. As a result, any potential to modify the physiology or behaviour of the host could play an important role in parasitoid fitness. The potential for host manipulation by the aphid parasitoid Aphidius avenae to increase cold thermotolerance was investigated using the aphid host species Metopolophium dirhodum and Sitobion avenae. Aphids were parasitized at L3/L4 instar stage (5 d old) and allowed to develop into pre-reproductive adults (10 d old) containing a 5 d old parasitoid larva. A control group was created of non-parasitized pre-reproductive adults (10 d old). The inherent physiological thermotolerance (LT50) and potential behavioural thermoregulation (behaviour in a declining temperature regime) of parasitized and non-parasitized aphids were investigated. Results revealed no effect of parasitism on the physiological thermotolerance of S. avenae and M. dirhodum. Significant differences in the behaviour of parasitized and non-parasitized aphids were observed, in addition to differences between host species, and such behaviours are discussed in view of the potential for host manipulation.

Could behaviour and not physiological thermal tolerance determine winter survival of aphids in cereal fields?

Traits of physiological thermotolerance are commonly measured in the laboratory as predictors of the field success of ectotherms at unfavourable temperatures (e.g. during harsh winters, heatwaves, or under conditions of predicted global warming). Due to being more complicated to measure, behavioural thermoregulation is less commonly studied, although both physiology and behaviour interact to explain the survival of ectotherms. The aphids Metopolophium dirhodum, Rhopalosiphum padi and Sitobion avenae are commercially important pests of temperate cereal crops. Although coexisting, these species markedly differ in winter success, with R. padi being the most abundant species during cold winters, followed by S. avenae and lastly M. dirhodum. To better understand the thermal physiology and behavioural factors contributing to differential winter success, the lethal temperature (physiological thermotolerance) and the behaviour of aphids in a declining temperature regime (behavioural thermotolerance) of these three species were investigated. Physiological thermotolerance significantly differed between the three species, with R. padi consistently the least cold tolerant and S. avenae the most cold tolerant. However, although the least cold tolerant of the study species, significantly more R. padi remained attached to the host plant at extreme sub-zero temperatures than S. avenae and M. dirhodum. Given the success of anholocyclic R. padi in harsh winters compared to its anholocyclic counterparts, this study illustrates that behavioural differences could be more important than physiological thermotolerance in explaining resistance to extreme temperatures. Furthermore it highlights that there is a danger to studying physiological thermotolerance in isolation when ascertaining risks of ectotherm invasions, the establishment potential of exotic species in glasshouses, or predicting species impacts under climate change scenarios.