Larval Viability Research Articles

EVALUATION OF BACTERIAL ISOLATES IN THE CONTROL OF Plutella xylostella L. IN BROCCOLI

<p>The diamondback moth (<em>Plutella xylostella</em> L.) is an insect that causes great losses in Brassica plantations, which may reach 100% loss in some cases. The most common method to control this pest is still the use of insecticide. However, its successive and arbitrary use has contributed to the development of insecticide resistance, and the study of alternative methods has become essential to successfully control the diamondback moth in broccoli. The objective of this study was to select isolates of rhizobacteria that act to control diamondback moth in broccoli. The experimental design was in randomized blocks, with three bacterial isolates [1 - <em>Kluyvera ascorbata</em> (EN4); 2 - <em>Bacillus subtllis</em> (R14); 3 - <em>Bacillus cereus</em> (C210) and 4 - Control (distilled water + spreader sticker)] tested with 5 repetitions in 20 m<sup>2</sup> plots. Larval viability (LV), pupal viability (PV), larval stage duration (LD), pupal stage duration (PD), pupal weight (PW) and total mortality (TM) were evaluated. The rhizobacteria isolates performed better to reduce larval and pupal viability, larval and pupal duration, and increase total mortality, and can be used as a management option to complement the control strategies of the diamondback moth in broccoli.</p>

Eficácia de ectoparasiticidas contra larvas de Aedes aegypti: nota de pesquisa

Foi avaliada a eficácia de produtos ectoparasiticidas contra larvas de Aedes aegypti. Os princípios ativos fluazuron 2,5 g, cipermetrina 150 g, flumetrina 1 g, propoxur 1 g, deltametrina 25 g, clorpirifós 30 g, cipermetrina 15 g, fenthion 15 g, diazinon técnico estabilizado 40 g, fipronil 2,5%, triclorfone 97 g, coumafós 3 g, propoxur 2 g, fentione 15 g e amitraz 12,5% foram adquiridos comercialmente e preparados conforme recomendações dos fabricantes. Os produtos foram distribuídos em recipientes individuais contendo larvas de A. aegypti em ambiente controlado. A inspeção da viabilidade larval foi verificada durante 24 horas. Os produtos foram aplicados nos locais de coleta das larvas (criadouros dos mosquitos), vistoriados diariamente por um mês. Foi observado que 100% das larvas morreram em até 60 minutos, tanto nos recipientes monitorados em laboratório como no meio ambiente. Nas áreas em que foi realizada a pulverização com os inseticidas supramencionados, não foi verificada a presença de larvas por um período de até 30 dias. Concluiu-se que produtos comumente utilizados em animais como ectoparasiticidas foram eficazes contra larvas de A. aegypti, podendo ser utilizados para o controle desse importante vetor, caso tal utilização seja analisada e regulamentada pelos órgãos oficiais do país.

Alternative to formaldehyde in artificial diet for sugarcane borer and its effects on the parasitoid

The formaldehyde used as anti-contaminant agent in artificial diets of insects can cause serious risks to human health. However, there are products with the same purpose, but with less toxicity to humans. In this sense, the aim of this study was to evaluate the biological parameters of Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) fed on an artificial diet containing different anti-infective agents to replace the formaldehyde, as well as analyze the effects on its parasitoid Cotesia flavipes (Cameron, 1891) (Hymenoptera: Braconidae). The experiment was conducted under a completely randomized design, studying the anti-contaminant agents: sodium benzoate, potassium sorbate, 2-phenylphenol, and formaldehyde (standard diet), with deionized water as control. The larvae of sugarcane borer were fed on diets with the cited agents, and were daily observed for: larval and pupal periods, larval and pupal viabilities, number of unviable larvae, longevity, pupae weight, sex ratio and the adults fecundity. To analyze the effects of the anti-contaminant agents on the C. flavipes parasitoid, 50 larvae of sugarcane borer were parasitized and daily observed until the parasitoid pupae formation, being evaluated: number of adults emerged per pupal mass, longevity and sex ratio. D. saccharalis larvae are positively influenced by replacing formaldehyde to 2-phenylphenol in their diet, not causing significant deleterious effects to C. flavipes.

Embryos of eastern rainbowfish Melanotaenia splendida splendida (Peters, 1866) tolerate fluctuating hypoxia

Hypoxia can have profound sublethal effects on reproduction and embryonic development of some freshwater fish. In the present study, the effects of diel fluctuating hypoxia on embryo viability were investigated for the eastern rainbowfish Melanotaenia splendida splendida, a small-bodied species common in wetlands of tropical Queensland. After daily hypoxic exposure (minimum 5% saturation) from fertilisation until hatch, no effects were found on egg incubation time, egg and larval mortality, and viability and size of hatching larvae. Older life history stages of the species are vulnerable to this level of hypoxia. Embryos of phytolithophilic species are likely exposed to fluctuating dissolved oxygen saturations in their natural habitat, and hypoxia tolerance may be a requirement for fish species that spawn predominantly on submerged plant material.

Effects of rhizobacteria on the biology and behavior of Plutella xylostella (Lepidoptera: Plutellidae)

Plant induced resistance is a tool for integrated pest management, aimed at increasing plant defense against stress, which is compatible with other techniques. Rhizobacteria act in the plant through metabolic changes and may have direct effects on plant-feeding insects. The objective of this study was to determine the effects of cabbage plants inoculated with rhizobacteria on the biology and behavior of diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae). Cabbage seeds inoculated with 12 rhizobacteria strains were sowed in polystyrene trays and later transplanted into the greenhouse. The cabbage plants with sufficient size to support stress were then infested with diamondback moth caterpillars. Later, healthy leaves suffering injuries were collected and taken to the laboratory to feed P. xylostella second instar caterpillars that were evaluated for larval and pupal viability and duration, pupal weight, and sex ratio. The reduction of leaf area was then calculated as a measure of the amount of larval feeding. Non-preference for feeding and oviposition assays were also performed, by comparing the control treatment and plants inoculated with different rhizobacterial strains. Plants inoculated with the strains EN4 of Kluyvera ascorbata and HPF14 of Bacillus thuringiensis negatively affected the biological characteristics of P. xylostella when such traits were evaluated together, without directly affecting the insect behavior.

Reproduction and embryo viability of a range-limited tropical freshwater fish exposed to fluctuating hypoxia

Hypoxia can profoundly affect fish reproduction and larval development, but its effects on fish from tropical Australia are not well understood. In the present study, the effects of diel fluctuating hypoxia on reproduction and embryo viability were investigated for a range-limited tropical freshwater fish, namely the Utchee Creek rainbowfish (Melanotaenia utcheensis). The lethal level for adult rainbowfish after gradual oxygen depletion was ~7% dissolved oxygen (DO) saturation. After 28 days, the reproductive success of adult fish exposed to fluctuating hypoxia treatments was measured by fecundity, gonad health, egg incubation time, egg and larval mortality, viability and size of hatching larvae. Reproduction was impaired in the lowest sublethal treatment (minimum 10% DO saturation each day). No ill effects of parental exposure to diel fluctuating hypoxia on embryos were identified, and minor differences in temperature between aquaria had a greater effect on embryos than parental hypoxia treatments. Similarly, no effects of embryonic exposure to diel fluctuating hypoxia were identified. Utchee Creek rainbowfish appear to be more hypoxia tolerant than temperate species, in keeping with their habitat in warm lowland streams, but they are still susceptible to the increasing frequency and intensity of hypoxia possible with increasing temperature and reduced flow as a result of climate change.

Biological aspects and predation behavior of Ceraeochrysa cubana against Spodoptera frugiperda

Mortality of neonate larvae and eggs of Spodoptera frugiperda has an important contribution to the management of this pest. This work aimed to determine the development, quantify the consumption, the time of search and handling of eggs of S. frugiperda by Cereaochrysa cubana. The duration of development and the survival of C. cubana were determined using eggs of the alternative prey Anagasta kuehniella; eggs, newborn caterpillars and combined feeding of egg + newborn caterpillars of S. frugiperda. The daily and total consumption, the search and handling time were also evaluated, comparing eggs and caterpillars of S. frugiperda as prey. The duration of the larvae to adult phase of C. cubana was prolonged when the food was newborn caterpillars or eggs + newborn caterpillars of S. frugiperda (35.7 and 34.6 days) when compared to eggs of A. kuehniella and S. frugiperda (25.5 and 25.9 days). The predator’s larvae showed a shorter larval and adult period when fed with eggs in relation to caterpillars. Larval viability of C. cubana was lower when fed with eggs + newborn caterpillars. Higher consumption and shorter search and handling time on newborn caterpillars of S. frugiperda, compared to prey eggs, are carried out by larvae of C. cubana during the third instar.

Assessing Common Bean Cultivars for Resistance to the Soybean Looper Chrysodeixis includens (Lepidoptera: Noctuidae).

The soybean looper Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae) is known as an important pest of leguminous plants worldwide. In Brazil, this pest species is gaining importance to producers of the common bean Phaseolus vulgaris L. (Fabaceae) because it limits field production of the crop. Chemical control is still the primary method of insect control. However, due to the possible harmful effects of pesticides to humans and the environment, alternative and less aggressive practices are being investigated. For this reason, the use of resistant plant genotypes represents a valuable tool in insect control. This study evaluated the biological aspects of larvae of C. includens confined to 14 bean genotypes under laboratory conditions (26±2°C; 65±10% RH; photoperiod of 14h L:10h D). The duration of the instars, total duration of the larval phase, consumption while in the larval phase, weight of the fifth instar larvae, larval viability, duration of the pre-pupal and pupal phases, pupal weight, pupal viability, pupal deformity, caterpillar-to-adult cycle, duration of the pre-oviposition and oviposition periods, and total number of viable eggs per female were evaluated. The genotypes "IAC Boreal," "IAC Harmonia," and "IAC Formoso" expressed antibiosis, prolonging the caterpillar-to-adult cycle and reducing the larval viability; however, each of these genotypes also experienced high leaf consumption. "IAC Jabola" expressed moderate levels of antibiosis and/or antixenosis (feeding), while the genotype "BRS Horizonte" expressed antixenosis (feeding). The data obtained with IAC Boreal, IAC Harmonia, IAC Formoso, IAC Jabola, and BRS Horizonte are promising and may help with the improvement of programs aimed at managing C. includens damage to this leguminous agricultural crop.

Production of diatom-bacteria biofilm isolated fromSeriola lalandicultures for aquaculture application

Controlled incorporation of selected microalgae and bacteria in aquaculture systems can be beneficial because they can act as microbiological control. That is why the characteristics of biofilm generated naturally in Seriola lalandi culture cages were analysed, their potential benefit to the growth of larvae was studied, and their controlled use for improving the larval viability and as a vector to improve incorporation of previously studied probiotic bacteria was tested. According to biodiversity results, these biofilms are composed of a diatom–bacteria mix showing a decrease in biodiversity in laboratory culture conditions being dominated by Navicula phyllepta and bacteria of the family Rhodobacteraceae. This can be produced on mesh substrates incorporated in bioreactors with rapid growth rate and adhesiveness. Preliminary results from the addition of substrates with this specific biofilm in larvae culture systems showed that it is consumed by the larvae without negative effects, while positive effects on the viability of larvae in combination with probiotics were observed. Considering preliminary results, the addition of these specific substrates with diatom–bacteria biofilms could be a good improvement for aquaculture systems and together with the use of probiotics can contribute to maintaining a stable and controlled system improving the viability of the larval fish culture in its early stages.

Response to rearing in laboratory of the xylophagous grape pest, Xylotrechus arvicola (Coleoptera: Cerambycidae)

AbstractXylotrechus arvicola (Olivier) (Coleoptera: Cerambycidae) is an important pest in vineyards (Vitis vinifera) in the main Iberian wine‐producing regions. Larvae were reared with Semi‐Synthetic Iglesias (SSI) diet over 27 months and two generations in the laboratory. Larval mortality was highest during the first (49.49 %) and second (9.38 %) month of rearing, increasing to 50.52 % during the first month if F2 reared larvae were obtained from an F1 adult female obtained in laboratory. The diet had sufficient nutrients to enable the pest to complete its life cycle within nine months, with F1 larval viability ranging from 23.49 % to 27.97 % and F2 larval viability reduced to 2.07 %. However, the diet did not allow for the completion of additional life cycles and generations (F3, F4,…). Larval mortality increased as the months of rearing (66.13 %, 69.51 % and 89.50 %) and generations (59.10 % and 76.93 % in F1 and F2, respectively) progressed in the laboratory. The larva–adult period of females obtained in the laboratory was longer than for males. In the laboratory, the life cycle was shortened in relation to the life cycle in the field because larvae did not require a cold period to break diapause and start pupation. This indicates that X. arvicola has the potential to complete its life cycle inside grape wood in vineyards of wine‐producing regions with warmer winters.

Selection and Suitability of an Artificial Diet for Tuta absoluta (Lepidoptera: Gelechiidae) Based on Physical and Chemical Characteristics.

Tuta absoluta (Meyrick, 1917) is a key tomato pest in South America and, recently, in Europe and Africa. To develop efficient control methods for this pest, adequate rearing protocols are desirable. As an alternative to tomato leaves (natural diet), we evaluated four artificial diets. Biological traits including larval and pupal viability and development time, pupal weight and deformations were assessed. Additionally, the optimum container size and larval density were evaluated. The diet based on casein, wheat germ and cellulose allowed the best development of T. absoluta, showing higher viability and no negative effects on larval instars and pupal weight. The best container was a glass tube measuring Ø 1 × h 6 cm, topped with waterproof cotton, with a density of three larvae. To evaluate the suitability of this diet, T. absoluta was reared during eight generations and life-table parameters were estimated for the F1, F3, F6, and F8 generations. The total viability (egg–adult) increased over the generations, reaching 75% in the eighth generation. Based on life-table estimations no differences among generations were found. The net reproductive rate (Ro) was higher than 40, the intrinsic rate of increase (rm) ranged between 0.08 and 0.11, the finite rate of increase (λ) was 1.1, the mean generation time (T) have a maximum of 44 d and doubling time ranged from 5.89–8.32 generations. These results indicated that a diet based on casein, wheat germ and cellulose was suitable for T. absoluta rearing in laboratory conditions.

A Theoretical Approach to Analyze the Parametric Influence on Spatial Patterns of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) Populations.

Studies of the influence of biological parameters on the spatial distribution of lepidopteran insects can provide useful information for managing agricultural pests, since the larvae of many species cause serious impacts on crops. Computational models to simulate the spatial dynamics of insect populations are increasingly used, because of their efficiency in representing insect movement. In this study, we used a cellular automata model to explore different patterns of population distribution of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), when the values of two biological parameters that are able to influence the spatial pattern (larval viability and adult longevity) are varied. We mapped the spatial patterns observed as the parameters varied. Additionally, by using population data for S. frugiperda obtained in different hosts under laboratory conditions, we were able to describe the expected spatial patterns occurring in corn, cotton, millet, and soybean crops based on the parameters varied. The results are discussed from the perspective of insect ecology and pest management. We concluded that computational approaches can be important tools to study the relationship between the biological parameters and spatial distributions of lepidopteran insect pests.

Coordinated inhibition of C/EBP by Tribbles in multiple tissues is essential for Caenorhabditis elegans development.

BackgroundTribbles proteins are conserved pseudokinases that function to control kinase signalling and transcription in diverse biological processes. Abnormal function in human Tribbles has been implicated in a number of diseases including leukaemia, metabolic syndromes and cardiovascular diseases. Caenorhabditis elegans Tribbles NIPI-3 was previously shown to activate host defense upon infection by promoting the conserved PMK-1/p38 mitogen-activated protein kinase (MAPK) signalling pathway. Despite the prominent role of Tribbles proteins in many species, our knowledge of their mechanism of action is fragmented, and the in vivo functional relevance of their interactions with other proteins remains largely unknown.ResultsHere, by characterizing nipi-3 null mutants, we show that nipi-3 is essential for larval development and viability. Through analyses of genetic suppressors of nipi-3 null mutant lethality, we show that NIPI-3 negatively controls PMK-1/p38 signalling via transcriptional repression of the C/EBP transcription factor CEBP-1. We identified CEBP-1’s transcriptional targets by ChIP-seq analyses and found them to be enriched in genes involved in development and stress responses. Unlike its cell-autonomous role in innate immunity, NIPI-3 is required in multiple tissues to control organismal development.ConclusionsTogether, our data uncover an unprecedented crosstalk involving multiple tissues, in which NIPI-3 acts as a master regulator to inhibit CEBP-1 and the PMK-1/p38 MAPK pathway. In doing so, it keeps innate immunity in check and ensures proper organismal development.Electronic supplementary materialThe online version of this article (doi:10.1186/s12915-016-0320-z) contains supplementary material, which is available to authorized users.

ZWEDGI: Wounding and Entrapment Device for Imaging Live Zebrafish Larvae.

Zebrafish, an established model organism in developmental biology, is also a valuable tool for imaging wound healing in space and time with cellular resolution. However, long-term imaging of wound healing poses technical challenges as wound healing occurs over multiple temporal scales. The traditional strategy of larval encapsulation in agarose successfully limits sample movement but impedes larval development and tissue regrowth and is therefore not amenable to long-term imaging of wound healing. To overcome this challenge, we engineered a functionally compartmentalized device, the zebrafish Wounding and Entrapment Device for Growth and Imaging (zWEDGI), to orient larvae for high-resolution microscopy, including confocal and second harmonic generation (SHG), while allowing unrestrained tail development and regrowth. In this device, larval viability was maintained and tail regrowth was improved over embedding in agarose. The quality of tail fiber SHG images collected from larvae in the device was similar to fixed samples but provided the benefit of time lapse data collection. Furthermore, we show that this device was amenable to long-term (>24 h) confocal microscopy of the caudal fin. Finally, the zWEDGI was designed and fabricated using readily available techniques so that it can be easily modified for diverse experimental imaging protocols.

Embryo-larval exposure to atrazine reduces viability and alters oxidative stress parameters in Drosophila melanogaster

The herbicide atrazine has been used worldwide with subsequent residual contamination of water and food, which may cause adverse effects on non-target organisms. Animal exposure to this herbicide may affect development, reproduction and energy metabolism. Here, the effects of atrazine regarding survival and redox metabolism were assessed in the fruit fly D. melanogaster exposed during embryonic and larval development. The embryos (newly fertilized eggs) were exposed to different atrazine concentrations (10μM and 100μM) in the diet until the adult fly emerged. Pupation and emergence rates, developmental time and sex ratio were determined as well as oxidative stress parameters and gene expression of the antioxidant defence system were evaluated in newly emerged male and female flies. Atrazine exposure reduced pupation and emergence rates in fruit flies without alterations to developmental time and sex ratio. Different redox imbalance patterns were observed between males and females exposed to atrazine. Atrazine caused an increase in oxidative damage, reactive oxygen species generation and antioxidant capacity and decreased thiol-containing molecules. Further, atrazine exposure altered the mRNA expression of antioxidant genes (keap1, sod, sod2, cat, irc, gss, gclm, gclc, trxt, trxr-1 and trxr-2). Reductions in fruit fly larval and pupal viability observed here are likely consequences of the oxidative stress induced by atrazine exposure.

Relationships between broodstock condition, oocyte quality, and 24 h D-larval survival during the spawning season of the pullet carpet shell Venerupis corrugata (Gmelin, 1791)

Venerupis corrugata is commercially exploited in Europe. Over-fishing and recruitment failure is causing the decline of its populations and stock sustainability. Knowledge of this species reproduction is paramount to establish hatchery production of juveniles for restoring natural beds. This work aimed to find a relationship between broodstock condition, oocyte quality, and viability of 24 h D-larvae. Adult specimens were induced to spawn by thermal stimulation. From each female, oocytes were taken for biochemical analyses (proteins, total lipids, and carbohydrates), and the remaining oocytes were fertilized. The 24 h D-larval yield was calculated after embryo incubation. Spawning in the hatchery with ‘wild’ broodstock was possible for a long period, however, subsequent larval viability varied according to oocyte quality. Two distinct periods of spawning were recorded: in January/March, with a higher number of oocytes released, and in June/July with a lower response to the spawning stimulation, however with greater success in 24 h D-larval survival. The condition index of broodstock and the total lipids of oocytes released can be used as benchmarks for estimating the success of D veliger larvae.

Toxic dinoflagellates and Vibrio spp. act independently in bivalve larvae

Harmful algal blooms (HABs) and marine pathogens – like Vibrio spp. – are increasingly common due to climate change. These stressors affect the growth, viability and development of bivalve larvae. Little is known, however, about the potential for interactions between these two concurrent stressors. While some mixed exposures have been performed with adult bivalves, no such work has been done with larvae which are generally more sensitive. This study examines whether dinoflagellates and bacteria may interactively affect the viability and immunological resilience of blue mussel Mytilus edulis larvae. Embryos were exposed to environmentally relevant concentrations (100, 500, 2500 & 12,500 cells ml−1) of a dinoflagellate (Alexandrium minutum, Alexandrium ostenfeldii, Karenia mikimotoi, Protoceratium reticulatum, Prorocentrum cordatum, P. lima or P. micans), a known pathogen (Vibrio coralliilyticus/neptunius-like isolate or Vibrio splendidus; 105 CFU ml−1), or both. After five days of exposure, significant (p < 0.05) adverse effects on larval viability and larval development were found for all dinoflagellates (except P. cordatum) and V. splendidus. Yet, despite the individual effect of each stressor, no significant interactions were found between the pathogens and harmful algae. The larval viability and the phenoloxidase innate immune system responded independently to each stressor. This independence may be related to a differential timing of the effects of HABs and pathogens.

Ekstrak Daun Pepaya dan Kangkung untuk Meningkatkan Daya Tetas Telur dan Kelangsungan Hidup Larva Lele (EXTRACTS OF CARICA PAPAYA AND IPOMOEA AQUATICA FOR IMPROVING EGG HATCHABILITY AND LARVAL VIABILITY OF CATFISH)

This research was aimed to investigate the potential use of leaf extract of Carica papaya and Ipomoeaaquatica lto improve egg hatchability and larval viability of catfish. Dried leaves of Carica papaya andIpomoea aquatica were macerated and extracted in water and ethanol. Eggs and larvae were tested in theaquarium size of 10 L with a a diameter of 28 cm. The extracts in concentration 600, 800 and 1.000 ppmwere tested on the egg hatchability of catfish with immersion method, and challed with Aeromonashydrophyla, Pseudomonas sp., and Saprolegnia spp. The extracts in concentration 800 and 1.000 ppm weretested on the larval viability with immersion method, and challed with pathogens. Water or ethanolextract of Carica papaya and Ipomoea aquatica can improve egg hatchability 67±8% until 90±6% andlarval viability of catfish 77±0,5 until 90±9%. Eight hundred ppm ethanol extract of Carica papaya has thebest egg hatchability and 1000 ppm can improve larval viability of catfish.

Redox imbalance mediates entomotoxic effects of the conifer Araucaria angustifolia in Anticarsia gemmatalis velvetbean caterpillar

AbstractThe velvetbean caterpillar, Anticarsia gemmatalis is one of the most important pests of soybean crops in tropical America. By feeding on leaves, significant defoliation occurs resulting in reduced photosynthetic capacity required for plants’ maintenance and growth, which subsequently can lead to crop losses and reduced agricultural productivity. Many studies have sought to look for compounds that have insecticidal effects. One class of compounds is phenolics, which are produced by plants and have been found to influence the behavior and development of defoliators, representing an important alternative approach to many synthetic insecticides. Particularly, Araucaria angustifolia is a plant rich in polyphenols, which are compounds able to alter cellular dynamics through modulating redox status. In this study, A. angustifolia extract (AAE) was added to the artificial diet of A. gemmatalis. The results demonstrated that AAE was able to reduce larval viability by inducing morphological changes and a de...

Efeito do extrato etanólico de sementes de Annona mucosa no desenvolvimento e comportamento alimentar de Spodoptera frugiperda

RESUMO No Brasil, a lagarta-do-cartucho Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) é uma praga importante do milho e de outras espécies cultivadas, incluindo cultivos hortícolas, soja, arroz e algodão. Inseticidas botânicos são considerados uma opção interessante para o manejo de S. frugiperda, especialmente em sistemas de produção orgânica, e derivados de Annonaceae constituem uma fonte promissora destes. Diante desse potencial, o efeito do extrato etanólico de sementes de Annona mucosa — ESAM (Annonaceae) no desenvolvimento e comportamento alimentar de S. frugiperda foi avaliado através de bioensaios de exposição em meio artificial (ingestão). ESAM reduziu a viabilidade das fases larval e pupal e o peso de pupa bem como provocou um aumento na duração da fase larval de S. frugiperda. Além disso, ESAM também causou um aumento na proporção de pupas e adultos com alterações morfológicas. Entretanto, ESAM não causou efeito deterrente de alimentação para lagartas de quarto ínstar de S. frugiperda, embora esse extrato tenha ocasionado um decréscimo significativo no consumo ao longo do tempo (toxicidade pós-ingestão).