A Time and Cost-Effective Pipeline for Expression Screening and Protein Production in Insect Cells Based on the HR-Bac Toolbox to Generate Recombinant Baculoviruses.

The baculovirus expression vector system (BEVS) is recognized as a powerful platform for producing challenging proteins and multiprotein complexes both in academia and industry. Since a baculovirus was first used to produce heterologous human IFN-ß protein in insect cells, the BEVS has continuously been developed and its applications expanded. We have recently established a multigene expression toolbox (HR-bac) composed of a set of engineered bacmids expressing a fluorescent marker to monitor virus propagation and a library of transfer vectors. Unlike platforms that rely on Tn7-medidated transposition for the construction of baculoviruses, HR-bac relies on homologous recombination, which allows to evaluate expression constructs in 2 weeks and is thus perfectly adapted to parallel expression screening. In this chapter, we detail our standard operating procedures for the preparation of the reagents, the construction and evaluation of baculoviruses, and the optimization of protein production for both intracellularly expressed and secreted proteins.

Assessment of the Entomopathogenic Potential of Fungal and Bacterial Isolates from Fall Armyworm Cadavers Against Spodoptera frugiperda Caterpillars and the Adult Boll Weevil, Anthonomus grandis.

Increased attention is being focused on the biological control of agricultural pests using microorganisms, owing to their potential as a viable substitute for chemical control methods. Insect cadavers constitute a potential source of entomopathogenic microorganisms. We tested whether bacteria and fungi isolated from Spodoptera frugiperda (JE Smith) cadavers could affect its survival, development, egg-laying pattern, and hatchability, as well as induce mortality in Anthonomus grandis Boheman adults. We isolated the bacteria Enterobacter hormaechei and Serratia marcescens and the fungi Scopulariopsis sp. and Aspergillus nomiae from fall armyworm cadavers and the pest insects were subjected to an artificial diet enriched with bacteria cells or fungal spores to be tested, in the case of S. frugiperda, and only fungal spores in the case of A. grandis. Enterobacter hormaechei and A. nomiae were pathogenic to S. frugiperda, affecting the survival of adults and pupae. The fungus Scopulariopsis sp. does not affect the survival of S. frugiperda caterpillars and pupae; however, due to late action, moths and eggs may be affected. Aspergillus nomiae also increased mortality of A. grandis adults, as well as the development of S. frugiperda in the early stages of exposure to the diet, as indicated by the vertical spore transfer to offspring and low hatchability. Enterobacter hormaechei and A. nomiae are potential biocontrol agents for these pests, and warrant further investigation from a toxicological point of view and subsequently in field tests involving formulations that could improve agricultural sustainability practices.

Thiadiazole/Thiadiazine Derivatives as Insecticidal Agent: Design, Synthesis, and Biological Assessment of 1,3,4-(Thiadiazine/Thiadiazole)-Benzenesulfonamide Derivatives as IGRs Analogues against Spodoptera littoralis.

In keeping with our investigation, a simple and practical synthesis of novel heterocyclic compounds with a sulfamoyl moiety that can be employed as insecticidal agents was reported. The compound 2-hydrazinyl-N-(4-sulfamoylphenyl)-2-thioxoacetamide 1 was coupled smoothly with triethylorthoformate or a variety of halo compounds, namely phenacyl chloride, chloroacetyl chloride, chloroacetaldehyde, chloroacetone, 1,3-dichloropropane, 1,2-dichloroethane, ethyl chloroformate, 2,3-dichloro-1,4-naphthoquinone, and chloroanil respectively, which afforded the 1,3,4-thiadiazole and 1,3,4-thiadiazine derivatives. The new products structure was determined using elemental and spectral analysis. Under laboratory conditions, the biological and toxicological effects of the synthetic compounds were also evaluated as insecticides against Spodoptera littoralis (Boisd.). Compounds 3 and 5 had LC50 values of 6.42 and 6.90 mg/L, respectively. The investigated compounds (from 2 to 11) had been undergoing molecular docking investigation for prediction of the optimal arrangement and strength of binding between the ligand (herein, the investigated compounds (from 2 to 11)) and a receptor (herein, the 2CH5) molecule. The binding affinity within docking score (S, kcal/mol) ranged between -8.23 (for compound 5), -8.12 (for compound 3) and -8.03 (for compound 9) to -6.01 (for compound 8). These compounds were shown to have a variety of binding interactions within the 2CH5 active site, as evidenced by protein-ligand docking configurations. This study gives evidence that those compounds have 2CH5-inhibitory capabilities and hence may be used for 2CH5-targeting development. Furthermore, the three top-ranked compounds (5, 3, and 9) and the standard buprofezin were subjected to density functional theory (DFT) analysis. The highest occupied molecular orbital-lowest unoccupied molecular orbital (HOMO-LUMO) energy difference (ΔE) of compounds 5, 3, and 9 was found to be comparable to that of buprofezin. These findings highlighted the potential and relevance of charge transfer at the molecular level.

Tetraniliprole risk assessment: Unveiling a hidden threat for managing a generalist herbivore.

Insecticide resistance poses a significant challenge in managing generalist herbivores such as the tobacco cutworm (TCW), Spodoptera litura. This study investigates the potential risks associated with using the novel diamide insecticide tetraniliprole to control TCW. A tetraniliprole-resistant strain was developed through twelve generations of laboratory selection, indicating an intermediate risk of resistance development. Field monitoring in China revealed a significant incidence of resistance, particularly in the Nanchang (NC) population (>100-fold). Tetraniliprole showed moderate to high cross-resistance to multiple insecticides and was autosomally inherited with incomplete dominance, controlled by multiple genes, some of which belong to the cytochrome P450 family associated with enhanced detoxification. Life table studies indicated transgenerational hormesis, stimulating TCW female fecundity and increasing population net reproduction rates (R0). These findings suggest a potential for pest resurgence under tetraniliprole use. The integrated risk assessment provides a basis for the sustainable management of TCW using tetraniliprole.

Characterization of Spodoptera littoralis (Lepidoptera: Noctuidae) resistance to indoxacarb: inheritance mode, realized heritability, and fitness costs.

Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae) is a major economic pest attacking a variety of crops in Egypt and other Mediterranean countries. S. littoralis has developed resistance to both traditional and novel insecticides. The current study investigated S. littoralis resistance to indoxacarb regarding inheritance mode, realized heritability (h2), and fitness costs. An indoxacarb-resistant strain (Indoxa-SEL) was obtained by selecting a field strain with indoxacarb. Indoxa-SEL strain outperformed the susceptible one (Indoxa-S) by 29.77-fold after 16 consecutive generations of selection. Based on the LC50 values of the progenies of reciprocal crosses F1 (R♂ × S♀) and F1' (R♀ × S♂), S. littoralis resistance to indoxacarb was found to be autosomal and partially recessive. Chi-square tests for goodness-of-fit between observed and expected mortalities of self-bred F1 and resistant strain reciprocal crosses revealed that the resistance was controlled by multiple genes. The resistant strain had a relative fitness of 0.80, with significantly increased total preovipositional period of females, egg, larvae, pupae, preadult, adult, and total longevity period. The estimated realized heritability value in the Indoxa-SEL strain was 0.21. The current study will contribute to sustaining indoxacarb efficacy and designing effective resistance management programs against S. littoralis.

Functional assessment of lysosomal Rab7 and RILP with RNA interference and overexpression in Spodoptera frugiperda Sf9 cell lines.

The interaction manner and biological function of Rab7 and its effector, Rab-interacting lysosomal protein (RILP), remain unclear in invertebrates. We provide a protocol for detecting the effects of Rab7 and RILP terminals on lysosome and autophagy in Spodoptera frugiperda Sf9 cells with overexpression and RNA interference. We describe steps for overexpressing plasmids, generating long double-stranded RNA, and transfecting them into Sf9 cells. We then detail procedures for cell immunofluorescence imaging with harmine treatment and fluorescence analysis. For complete details on the use and execution of this protocol, please refer to Cui et al. (2023).1.

Biology and quality assessment of Telenomus remus (Hymenoptera: Scelionidae) and Trichogramma spp. (Hymenoptera: Trichogrammatidae) in eggs of Spodoptera spp. for augmentative biological control programs.

The Spodoptera complex of the family Noctuidae, represented here by S. frugiperda (J.E. Smith), S. eridania (Stoll), S. albula (Walker), and S. cosmioides (Walker), is an important group of crop pests in Brazil. Spodoptera frugiperda and S. eridania are invasive in Africa, and the former also in Asia and Oceania. The egg parasitoids Telenomus remus Nixon (Hymenoptera: Scelionidae) and Trichogramma spp. (Hymenoptera: Trichogrammatidae) are potential control agents for field use against these noctuids. We evaluated the parasitism efficiency, development, and flight capacity of an isofemale line and a regular line of T. remus, and 2 genetically variable populations of Trichogramma pretiosum Riley and Trichogramma atopovirilia Oatman and Platner (Hymenoptera: Trichogrammatidae) in these 4 members of the Spodoptera complex. All parasitoids were able to develop in the 4 hosts. The parasitoids showed good flight capacity, except for the regular line of T. remus. The Trichogramma species, despite having high viability and female:male sex ratios, showed poorer parasitism performances than T. remus. The regular T. remus line also showed good parasitism capacity and high viability but had a predominance of males. In general, the isofemale line of T. remus showed good rates of parasitism and flight capacity as well as a high viability and sex ratio, proving to be a potential candidate for an augmentative biological-control program for Spodoptera spp Guenée (Lepidoptera: Noctuidae).

Quality assessment of Telenomus remus successively reared on Spodoptera litura eggs for 30 generations.

BACKGROUND: Telenomus remus (Nixon) is a dominant natural enemy controlling the invasive pest Spodoptera frugiperda (J. E. Smith). Continuous rearing of egg parasitoids on alternative hosts is crucial for mass production and cost reduction. However, to ensure the effectiveness of natural enemy products against target pests in the field, it is necessary to evaluate the parasitoid quality during the mass-rearing process. Despite the successful rearing of this parasitoid on the alternative host Spodoptera litura (Fabricius) eggs, less attention has been paid to the quality of parasitoids continuously reared for multiple generations. Therefore, we evaluated the performance of T. remus reared on S. litura eggs for 30 generations via morphological characteristics, flight ability, and life table analysis. RESULTS: Wing length, wing width, body length, and right hind tibia length of T. remus did not differ among the different generations. However, the body length of female parasitoids was significantly longer than that of males for any generation. Although the proportion of 'flyers' and 'deformed' T. remus varied among generations, the flight ability did not decline significantly after rearing on S. litura eggs. Moreover, T. remus continuously reared on S. litura eggs maintained stable parasitism performance and life table parameters on the target host S. frugiperda eggs. CONCLUSION: S. litura eggs are suitable hosts for the mass-rearing of T. remus. This study can be subsequently used to guide the production and facilitate the application of T. remus in the control of S. frugiperda. © 2023 Society of Chemical Industry.

The Cost of Resistance to Diamide Insecticide Varies With the Host Plant in Spodoptera frugiperda (Lepidoptera: Noctuidae).

Fitness costs associated with insect resistance to insecticides can be exploited to implement resistance management programs. However, most of these studies are restricted to evaluating biological traits on artificial diets. Here, we investigated the fitness cost associated with chlorantraniliprole in Spodoptera frugiperda (J.E. Smith) feeding on corn, soybean, and cotton plants. We used a near-isogenic strain of S. frugiperda resistant to chlorantraniliprole (Iso-RR), a susceptible strain (SS), and heterozygotes strains (H1 and H2) to evaluate several biological and population growth parameters. Larval survival of the Iso-RR strain was on average 90% on corn, 65% on soybean, and 57% on cotton plants. Development time of the larval stage also differed among host plants, Iso-RR strain took on average 14, 17, and 26 days to reach the pupal stage on corn, soybean, and cotton plants respectively. Net reproductive rate, intrinsic rate of population increase, and finite rate of population increase were higher for Iso-RR strain feeding on corn plants than other host plants. The relative fitness, based on the intrinsic rate of population increase, of S. frugiperda resistant strain on corn, soybean, and cotton plants were 1.04, 0.85, and 0.88, respectively. Therefore, no fitness cost was observed for S. frugiperda feeding on corn plants, but a significant fitness cost was observed when this pest fed on soybean and cotton plants. We showed that the food source influences the fitness cost of S. frugiperda resistant to diamide. Such information may help to implement resistance management strategies based on each crop.

A cost-efficient and alternative technique of managing fall armyworm Spodoptera frugiperda (J.E. Smith) larvae in maize crop.

An experiment was conducted to test the efficacy of grease and emamectin benzoate in a randomized complete block design with five replications to reduce fall armyworm, Spodoptera frugiperda (J.E. Smith) larvae load from a maize field in the winter seasons of 2020 and 2021 in Sarlahi, Nepal. Standard agronomic package of practices followed for crop proper growth and development, and plant spacing maintained at 20 × 60  cm2 with a plot size of 390 m2. The treatments were applied when the maize crop was at knee height and larvae damaged nearly 5-8% of the total plants. Emamectin benzoate sprayed at 0.4 g/liter of water and grease of about 0.15 g applied to the maize whorl or tip of a drooping leaf that touched the soil. A significant reduction in larval infestation was observed after 7-days of treatment applications. Fall armyworm larvae were found dead in the chemical-sprayed plots, but they were absent in the grease-applied fields. No crop damage was observed among the grease-treated plants, which might be due to restrictions in the movement of larvae on the maize crop. The armyworm larvae might get irritated, feel insecure, and move far away from the test plots searching for food materials. Thus, an eco-friendly material like grease can be used as an agroecological method for managing fall armyworm larvae among small-scale land-holding maize farmers.

Assessing fitness costs of the resistance of Spodoptera frugiperda (Lepidoptera: Noctuidae) to pyramided Cry1 and Cry2 insecticidal proteins on different host plants.

Fall armyworm (FAW), Spodoptera frugiperda (Smith), is one of the major pests targeted by transgenic crops expressing insecticidal proteins from Bacillus thuringiensis (Bt) Berliner. However, FAW presents a high capacity to develop resistance to Bt protein-expressing crop lines, as reported in Brazil, Argentina, Puerto Rico and the southeastern U.S. Here, FAW genotypes resistant to pyramided maize events expressing Cry1F/Cry1A.105/Cry2Ab2 (P-R genotype) and Cry1A.105/Cry2Ab2 (Y-R genotype) from Brazil were used to investigate the interactions between non-Bt hosts (non-Bt maize, non-Bt cotton, millet and sorghum) and fitness costs. We also tested a FAW genotype susceptible to Bt maize and F1 hybrids of the resistant and susceptible genotypes (heterozygotes). Recessive fitness costs (i.e., costs affecting the resistant insects) were observed for pupal and neonate to adult survival of the P-R genotype on non-Bt cotton; larval developmental time of the P-R genotype on millet and sorghum; larval and neonate-to-adult developmental time of the Y-R genotype on non-Bt cotton and sorghum; the fecundity of the Y-R genotype on non-Bt cotton; and mean generation time of both resistant genotypes. However, on non-Bt cotton and non-Bt maize, the P-R genotype had a higher fitness (i.e., fitness benefits), displaying greater fecundity and rates of population increases than the Sus genotype. Non-recessive fitness costs (i.e., costs affecting heterozygotes) were found for fecundity and population increases on millet and sorghum. These findings suggest that, regardless of the disadvantages of the resistant genotypes in some hosts, the resistance of FAW to Cry1 and Cry2 Bt proteins is not linked with substantial fitness costs, and may persist in field conditions once present.

Fitness costs associated with chlorantraniliprole resistance in Spodoptera frugiperda (Lepidoptera: Noctuidae) strains with different genetic backgrounds.

BACKGROUND: Spodoptera frugiperda (J.E. Smith) is a difficult pest to manage mainly because of its resistance to insecticides and Bt proteins. We evaluated fitness costs of S. frugiperda resistant strains to diamide insecticides with different genetic backgrounds aiming to highlight the importance of using isogenic strains. We established a near-isogenic strain of S. frugiperda resistant to diamides (Iso-RR), using a chlorantraniliprole resistant strain (RR) selected from a field-collected population and a susceptible reference strain (SS). Fitness costs were assayed using strains with close-related genetic backgrounds (Iso-RR and SS) and strains with distant-related genetic backgrounds (RR and SS). RESULTS: No fitness cost associated with chlorantraniliprole resistance in S. frugiperda was observed using the Iso-RR strain, based on life history traits. The only parameter that differs between Iso-RR and SS strains was the mean length of a generation (T), whereas the Iso-RR strain presented T = 35.8 and SS strain showed T = 34.6. On the other hand, a significant fitness cost was detected using the RR strain. All population growth parameters differ between RR and SS strains. Based on the intrinsic rate of population increase (rm ) parameter, the relative fitness estimated was 1.02 for the Iso-RR strain and 0.64 for the RR strain. CONCLUSION: The genetic background of the resistant strains alters fitness cost outcomes. The RR strain showed fitness costs associated with resistance, but the Iso-RR did not. Our work supports the decision-making process of resistance management programs and adds to the growing body of research that enlightens the importance of strain genetics in fitness cost experiments.

Socioeconomic and health impacts of fall armyworm in Ethiopia.

Since 2016, fall armyworm (FAW) has threatened sub-Saharan 'Africa's fragile food systems and economic performance. Yet, there is limited evidence on this transboundary pest's economic and food security impacts in the region. Additionally, the health and environmental consequences of the insecticides being used to control FAW have not been studied. This paper presents evidence on the impacts of FAW on maize production, food security, and human and environmental health. We use a combination of an agroecology-based community survey and nationally representative data from an agricultural household survey to achieve our objectives. The results indicate that the pest causes an average annual loss of 36% in maize production, reducing 0.67 million tonnes of maize (0.225 million tonnes per year) between 2017 and 2019. The total economic loss is US$ 200 million, or 0.08% of the gross domestic product. The lost production could have met the per capita maize consumption of 4 million people. We also find that insecticides to control FAW have more significant toxic effects on the environment than on humans. This paper highlights governments and development partners need to invest in sustainable FAW control strategies to reduce maize production loss, improve food security, and protect human and environmental health.

Pathogenic Assessment of SfMNPV-Based Biopesticide on Spodoptera frugiperda (Lepidoptera: Noctuidae) Developing on Transgenic Soybean Expressing Cry1Ac Insecticidal Protein.

Pathogenic assessment of a baculovirus-based biopesticide containing Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV: Baculoviridae: Alphabaculovirus) infecting fall armyworm, Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae) is reported. In the bioassays, neonates were infected with different doses of SfMNPV applied on Cry1Ac Bt soybean and non-Bt soybean. Our findings indicated that S. frugiperda neonates did not survive at 10 d post infection or develop into adults on Bt and non-Bt soybean sprayed with the field recommended dose of SfMNPV. In contrast, a proportion of the infected neonates developed into adults when infected with lower doses of SfMNPV (50%, 25%, and 10% of field dose) in both Bt and non-Bt soybean. However, S. frugiperda neonates surviving infection at the lowest virus doses on both soybean varieties showed longer neonate-to-pupa and neonate-to-adult periods, lower larval and pupal weights, reduced fecundity, and increased population suppression. Nevertheless, more pronounced pathogenicity of SfMNPV infecting neonates of S. frugiperda were verified on larvae that developed on Bt soybean. These findings revealed that, beyond mortality, the biopesticide containing SfMNPV also causes significant sublethal pathogenic effects on neonates of S. frugiperda developing on Bt and non-Bt soybean and suggested an additive effect among SfMNPV and Cry1Ac insecticidal protein expressed in Bt soybean.

Rapid and cost-effective process based on insect larvae for scale-up production of SARS-COV-2 spike protein for serological COVID-19 testing.

Serology testing for COVID-19 is important in evaluating active immune response against SARS-CoV-2, studying the antibody kinetics, and monitoring reinfections with genetic variants and new virus strains, in particular, the duration of antibodies in virus-exposed individuals and vaccine-mediated immunity. In this study, recombinant S protein of SARS-CoV-2 was expressed in Rachiplusia nu, an important agronomic plague. One gram of insect larvae produces an amount of S protein sufficient for 150 determinations in the ELISA method herein developed. We established a rapid production process for SARS-CoV-2 S protein that showed immunoreactivity for anti-SARS-CoV-2 antibodies and was used as a single antigen for developing the ELISA method with high sensitivity (96.2%) and specificity (98.8%). Our findings provide an efficient and cost-effective platform for large-scale S protein production, and the scale-up is linear, thus avoiding the use of complex equipment like bioreactors.

Fitness Costs of Chlorantraniliprole Resistance Related to the SeNPF Overexpression in the Spodoptera exigua (Lepidoptera: Noctuidae).

Spodopteraexigua, a multifeeding insect pest, has developed a high level of resistance to chlorantraniliprole, which is a benzoylurea insecticide that targets the ryanodine receptors (RyRs). Herein, the resistant strain (SE-Sel) and sensitive strain (SE-Sus) were obtained by bidirectional screening for six generations. The potential oviposited eggs and oviposition rate of the SE-Sel strain were dramatically lower than those of the SE-Sus strain; on the contrary, the weights of prepupae and preadult were significantly increased. As a post-mating response, the higher number of non-oviposited eggs in the SE-Sel strain was caused by a lower mating rate. In addition, the expression levels of vitellogenin (SeVg) and its receptor (SeVgR) in the SE-Sel strain were consistently lower than those in the SE-Sus strain. An RyRI4743M mutation, contributing to the resistance to chlorantraniliprole, was located in the S3 transmembrane segments and might have affected the release of calcium ions; it led to the upregulated expression of the neuropeptide SeNPF and its receptor SeNPFR, and the mating and oviposition rate were significantly recovered when the SeNPF was knocked down though RNA interference (RNAi) in the male adult of the SE-Sel strain. Moreover, the expression of the juvenile hormone-binding proteins SeJHBWDS3 and SeJHBAN in the male adult of the SE-Sel strain was significantly decreased, which proved the existence of a fitness cost from another angle. Therefore, these results indicate that the fitness cost accompanied by chlorantraniliprole resistance in S. exigua may be related to the decrease in mating desire due to SeNPF overexpression.

What Will Fall Armyworm (Lepidoptera: Noctuidae) Cost Western Australian Agriculture?

Following the detection of fall armyworm Spodoptera frugiperda (J.E. Smith, Lepidoptera: Noctuidae) in Western Australia in early 2020 and the lack of government response action, we estimate the impact it is likely to have on the state's agriculture. A bioeconomic model is used to estimate cost and revenue implications for broadacre cropping and horticulture industries. We assume permanent S. frugiperda populations are likely to establish in areas of the state's north and mid-west over the next decade, and other regions may experience sporadic outbreaks over single seasons. Over 0.8 million hectares of host crops could be permanently affected, while sporadic outbreaks may affect a further 150,000 hectares. Expressed in Australian dollars (A$), S. frugiperda is likely to add a A$14.2-39.3 million burden to agricultural producers per annum by year 10 of the outbreak. Approximately 55% of these damage costs are attributable to yield loss and 45% to increased variable production costs.

Fitness Cost of Chlorpyrifos Resistance in Spodoptera frugiperda (Lepidoptera: Noctuidae) on Different Host Plants.

Spodoptera frugiperda (J. E. Smith, 1797) is a polyphagous pest of global relevance due to the damage it inflicts on agricultural crops. In South American countries, this species is one of the principal pests of maize and cotton. Currently, S. frugiperda is also emerging as an important pest of soybeans and winter cereals in Brazil. Chemical control is one of the main control tactics against S. frugiperda, even though resistance against numerous modes of action insecticides has been reported. To support insect resistance management programs, we evaluated the fitness costs of resistance of S. frugiperda to the acetylcholinesterase inhibitor chlorpyrifos. Fitness costs were quantified by comparing biological parameters of chlorpyrifos-resistant and -susceptible S. frugiperda and their F1 hybrids (heterozygotes) on non-Bt cotton, non-Bt maize, non-Bt soybean, and oats. The results revealed that the chlorpyrifos-resistant genotype showed lower pupa-to-adult and egg-to-adult survivorship and reduced larval weights on oats; longer neonate-to-pupa and egg-to-adult developmental periods, and lower pupal weights and fecundity on maize; lower pupal weights on soybean; and reduced fecundity on cotton compared with the chlorpyrifos-susceptible genotype. Fitness costs also affected fertility life table parameters of the resistant genotype, increasing the mean length of a generation on cotton and maize and reducing the potential for population growth on all hosts. These findings suggest fitness costs at the individual and population levels of chlorpyrifos resistance in S. frugiperda, indicating that removal of the selective agent from the environment would result in reduced resistance and opportunities for the restoration of susceptibility.

Fall armyworm invasion heightens pesticide expenditure among Chinese smallholder farmers.

Invasive species are a prominent feature of global change. Aside from their direct impacts on biodiversity and ecosystem functioning, invasive crop pests routinely trigger environmentally-disruptive actions e.g., unguided applications of synthetic pesticides. Since 2016, the polyphagous fall armyworm (FAW, Spodoptera frugiperda J.E. Smith) has rapidly spread across Africa and Asia, impacting millions of hectares of agricultural crops. Upon its invasion of Yunnan (China) in late 2018, S. frugiperda attained outbreak population levels and inflicted important feeding damage in smallholder-managed maize crops. In this study, we show how local maize growers rely primarily on pesticides for FAW management and employ these products at 3-fold higher application frequencies as compared to 2018. Local reliance upon high-risk compounds (i.e., pyrethroids, organophosphates) decreased over time, with a respective 100% and 62% farmers using these compounds in 2018 versus 27% and 5% in 2020. Conversely, 71% and 95% farmers used new, selective compounds such as emamectin benzoate and chlorfenapyr by 2020. The full cost of pesticide-based crop protection increased from US $81 per hectare and season in 2018 to $276 in 2020. In farmer-managed fields, FAW infestation levels averaged 8.3 larvae per 100 plants and thus remained below economic injury levels (EILs) as established in other countries. Farmers' use of two or more pesticide sprays per season likely was not economically justified. Our work demonstrates how the FAW invasion has altered pest management regimes in Yunnan's maize crop, deepening farmers' pesticide dependency, and potentially exacerbating its burden on household budgets. Sustainable pest management schemes urgently need to be devised for smallholder maize systems in China and across the FAW invaded range.

Fitness costs associated with chlorantraniliprole resistance in Spodoptera exigua (Lepidoptera: Noctuidae).

BACKGROUND: The field population of Spodoptera exigua, an intermittently occurring polyphagous pest, has developed resistance to chlorantraniliprole, while whether or not such resistance carries fitness costs remains poorly understood. Here we selected six generations of the Leshan population (LS-P) by two-way selecting method, and obtained a highly resistant strain (CH-RE) and resistant degeneration strain (CH-SE) sharing a similar genetic background. After that fitness costs were evaluated by comparing the life history characteristics of CH-RE, CH-SE and the laboratory susceptible strain (SE-Lab) via the age-stage two-sex life table method. RESULTS: The resistance ratio of CH-RE and CH-SE were 226.69-fold and 3.72-fold, respectively, and the estimated realized heritability (h2 ) of CH-RE was 0.058. Compared with CH-SE, the duration of pre-adult, the longevity of adult, adult preoviposition period (APOP) and average generation time (T) of CH-RE had significantly increased, but the oviposition days, average fecundity, intrinsic growth rate (r), weekly growth rate (λ) and reproductive rate (R0 ) decreased significantly. Moreover, the relative fitness of CH-RE was 0.25, and showed fitness costs. Concurrently, the fecundity of CH-SE was slightly lower than SE-Lab, but there was no significant difference in r, λ and R0 , and the fitness (1) of CH-SE was similar to SE-Lab (1.02), which was no fitness cost. CONCLUSION: These findings represent that chlorantraniliprole resistance in S. exigua has a fitness cost, and the fitness cost will disappear with the recovery of sensitivity when the insecticide is stopped for field populations, supporting that such resistance would be managed by switching off the selection pressure with rotation with alternate insecticides. © 2020 Society of Chemical Industry.