Larval Mortality Research Articles

Preferences for livestock bedding as a development substrate of the stable fly, Stomoxys calcitrans L. (Diptera: Muscidae), and potential application of entomopathogenic nematodes for controlling stable fly larvae.

The stable fly, Stomoxys calcitrans L. (Diptera: Muscidae), is a significant insect pest with global veterinary implications due to its capacity to both cause nuisance and transmit disease-causing pathogens to livestock. This study aimed to determine the livestock bedding preferred for use as a development substrate by S. calcitrans larvae and field-collected adults. The result showed that S. calcitrans larvae exhibited a preference (26.7%) for 7-day-old cow manure. Gravid females displayed a pronounced preference (55.0%) for fresh cow manure. As there were eight choices, indifference would result in 12.5% for each bedding substrate. Furthermore, the efficacy of four entomopathogenic nematodes (EPNs), namely Heterorhabditis bacteriophora (Poinar), Heterorhabditis indica Poinar, Karunakar & David (Rhabditida: Heterorhabditidae), Steinernema siamkayai Poinar, Karunakar & David and Steinernema carpocapsae (Weiser) (Rhabditida: Steinernematidae), against S. calcitrans larvae and the persistence after application to livestock bedding substrates were evaluated under laboratory conditions. In filter paper bioassays, all four EPNs caused 76.7%-100.0% mortality in the second instar larvae of S. calcitrans when applied at 50 and 100 infective juveniles (IJs)/cm2 within 5 days after exposure. For the third instar larvae of S. calcitrans, only H. indica induced high mortalities of 86.6% when applied at 100 IJs/cm2 within 5 days after exposure, while the other EPNs resulted in mortalities of less than 70%. The data further demonstrated that H. bacteriophora, H. indica and S. siamkayai remained present in the substrates linked to S. calcitrans larvae for as long as 7 days after the application of EPNs. This study demonstrates the potential of EPNs as a biologically based control agent against larvae of S. calcitrans, a serious pest and significant vector for various livestock animals.

Insecticidal activity of Baccharis essential oils against Duponchelia fovealis, toxicity to strawberry leaves, and in vitro biocompatibility with entomopathogenic fungi

ABSTRACT Duponchelia fovealis is responsible for great losses in the strawberry production, and entomopathogenic fungi (EPF) are an alternative to control this larva. They can be used in combination with other natural compounds with insecticidal activity, such as plant extracts or essential oils (EOs). Thus, our objectives are to evaluate the effects of Baccharis dracunculifolia and Baccharis uncinella EOs on strawberry leaves and third instar larvae of D. fovealis, and to evaluate their in vitro compatibility with strains of Beauveria bassiana and Cordyceps javanica. EOs were not phytotoxic to strawberry leaves at 2% v/v. Larval mortality ranged between 48% and 83% at 1% and 2%, and B. dracunculifolia EO at 2% presented the higher mortality. EO of B. uncinella (BU) at 1% was compatible with two strains of B. bassiana and C. javanica. Therefore, the combined use of BU EO with EPF may be an alternative to control D. fovealis.

Assessment of biorational larvicides and botanical oils against Culex quinquefasciatus Say (Diptera: Culicidae) larvae in laboratory conditions

Mosquitoes are known vectors that transmit deadly diseases to millions of people across the globe. The reliance on synthetic insecticides has been the sole way to combat mosquito vectors for decades. In recent years, the extensive use of conventional insecticides in mosquito suppression has led to significant pesticide resistance and serious human health hazards. In this light, investigating the potential application of biorational compounds for vector management has drawn significant attention. We, hereby, evaluated the efficacy of three microbial derivative biorational insecticides, abamectin, spinosad, and buprofezin, and two botanical oils, neem (Azadirachta indica A. Juss) and karanja oil (Pongamia pinnata Linn.) against the Culex quinquefasciatus under laboratory conditions. The fourth-instar C. quinquefasciatus larvae were exposed to different concentrations of the selected larvicides and lethality was estimated based on LC50 and LT50 with Probit analysis. All larvicides showed concentration-dependent significant effects on survival and demonstrated larvicidal activity against C. quinquefasciatus larvae. However, abamectin exerted the highest toxicity (LC50 = 10.36 ppm), exhibited statistically significant effects on C. quinquefasciatus larval mortality, followed by spinosad (LC50 = 21.32 ppm) and buprofezin (LC50 = 56.34 ppm). Abamectin caused larval mortality ranged from 30.00 to 53.33 % and 53.00–70.00 % at 06 and 07 h after treatment (HAT), respectively. In the case of botanicals, karanja oil (LC50 = 216.61 ppm) was more lethal (more than 1.5 times) and had a shorter lethal time than neem oil (LC50 = 330.93 ppm) and showed a classic pattern of relationship between concentrations and mortality over time. Overall, the present study highlighted the potential of deploying new generation biorational pesticides and botanicals in mosquito vector control programs.

Efektivitas Biolarvasida Kombinasi Ekstrak Daun Sirsak (Annona muricata) dan Bunga Marigold (Tagetes erecta L.) sebagai Pembasmi Jentik Nyamuk

This research aims to determine the effectiveness of using a biolarvicide combination of soursop leaf extract and marigold flowers as an exterminator for mosquito larvae. This research included a laboratory experiment using a completely randomized 2 factorial design. The first factor is a combination of soursop leaf extract and marigold flowers, namely, a) 75%:25% (F1); b) 50%:50% (F2); c) 25%:75% (F3). The second factor is the treatment dose, namely, a) 1% (D1); b) 2% (D2); c) 2.5% (D3). Each factor was repeated 3 times. Data analysis uses a non-parametric test, namely Kruskal-Wallist. The results of the study showed that the soursop leaf and marigold flower extract formula 75%:25% was most effective against mosquito larvae mortality at a dose of 2.5% within an observation period of 6 hours. In conclusion, soursop leaf and marigold flower extracts are effective mosquito larva repellents. Keywords: Biolarvicide, Marigold Flowers, Soursop Leaves, Effectiveness, Mosquito Larvae

Toxicity of Bt Protein on Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) Fed With Corn Leaves in Paraguay

The fall armyworm Spodoptera frugiperda is the most destructive pest in corn cultivation in Paraguay with yield losses of 20 to 70%. The adoption of genetically modified crops (GMO) is a valid technique for managing this pest. The objective of this investigation was to evaluate the toxicity among Bt events used locally; determining mortality and important biological parameters. The S. frugiperda population previously collected from the field was raised under laboratory conditions fed with leaves from events MIR162TM (Vip3A19), VT3ProTM (Cry1A105, Cry2Ab2, Cry3Bb), PowerCoreTM (Cry1A105, Cry1F, Cry2Ab2) and a treatment with non-maize. Bt BR106TM. The mortality of 120 larvae was evaluated for each treatment and the viability until the adult stage of those larvae that managed to survive the different treatments was observed. A high toxicity effect was verified with 100% mortality of neonatal larvae fed with the MIR162TM (Vip3A19) event; as for the other events, 86% and 61% VT3ProTM (Cry1A105, Cry2Ab2, Cry3Bb), and PowerCoreTM (Cry1A105, Cry1F, Cry2Ab2) respectively; with 13% mortality for the non-Bt control variety BR106TM. The oviposition period, viability and number of eggs laid in adults surviving the PowerCoreTM (Cry1A105, Cry1F, Cry2Ab2) and VT3ProTM (Cry1A105, Cry2Ab2, Cry3Bb) events were reduced, affecting the fecundity of the surviving insect population.

Toxicity of Bt Protein on Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) Fed With Corn Leaves in Paraguay

The fall armyworm Spodoptera frugiperda is the most destructive pest in corn cultivation in Paraguay with yield losses of 20 to 70%. The adoption of genetically modified crops (GMO) is a valid technique for managing this pest. The objective of this investigation was to evaluate the toxicity among Bt events used locally; determining mortality and important biological parameters. The S. frugiperda population previously collected from the field was raised under laboratory conditions fed with leaves from events MIR162TM (Vip3A19), VT3ProTM (Cry1A105, Cry2Ab2, Cry3Bb), PowerCoreTM (Cry1A105, Cry1F, Cry2Ab2) and a treatment with non-maize. Bt BR106TM. The mortality of 120 larvae was evaluated for each treatment and the viability until the adult stage of those larvae that managed to survive the different treatments was observed. A high toxicity effect was verified with 100% mortality of neonatal larvae fed with the MIR162TM (Vip3A19) event; as for the other events, 86% and 61% VT3ProTM (Cry1A105, Cry2Ab2, Cry3Bb), and PowerCoreTM (Cry1A105, Cry1F, Cry2Ab2) respectively; with 13% mortality for the non-Bt control variety BR106TM. The oviposition period, viability and number of eggs laid in adults surviving the PowerCoreTM (Cry1A105, Cry1F, Cry2Ab2) and VT3ProTM (Cry1A105, Cry2Ab2, Cry3Bb) events were reduced, affecting the fecundity of the surviving insect population.

Degradation mechanism and decomposition of sulfamethoxazole aqueous solution with persulfate activated by dielectric barrier discharge

ABSTRACT The present study focused on the degradation of sulfamethoxazole (SMX) aqueous solution and the toxicity of processing aqueous by the dielectric barrier discharge (DBD) activated persulfate (PS). The effects of input voltage, input frequency, duty cycle, and PS dosage ratio on the SMX degradation efficiency were measured. Based on the results of the Response Surface Methodology (RSM), SMX degradation efficiency reached 83.21% which is 10.54% higher than that without PS, and the kinetic constant was 0.067 min−1 in 30 min when the input voltage at 204 V (input power at 110.6 W), the input frequency at 186 Hz, the duty cycle at 63%, and the PS dosage ratio at 5.1:1. The addition of PS can produce more active particles reached 1.756 mg/L (O3), 0.118 mg/L (H2O2), 0.154 mmol/L (·OH) in 30 min. Furthermore, the DBD plasma system effectively activated an optimal amount of PS, leading to improved removal efficiency of COD, and TOC to 30.21% and 47.21%, respectively. Subsequently, eight primary by-products were pinpointed, alongside the observation of three distinct pathways of transformation. Predictions from the ECOSAR software indicated that most of the degradation intermediates were less toxic than SMX. The biological toxicity experiments elucidated that the treatment with the DBD/PS system effectively reduced the mortality of zebrafish larvae caused by SMX from 100% to 20.13% and improved the hatching rate from 55.69% to 80.86%. In particular, it is important to note that the degradation intermediates exhibit teratogenic effects on zebrafish larvae.

GABA attenuates neurotoxicity of zinc oxide nanoparticles due to oxidative stress via DAF-16/FoxO and SKN-1/Nrf2 pathways

Zinc oxide nanoparticles (ZnO-NPs) are one of the most widely used metal oxide nanomaterials. The increased use of ZnO-NPs has exacerbated environmental pollution and raised the risk of neurological disorders in organisms through food chains, and it is urgent to look for detoxification strategies. γ-Aminobutyric acid (GABA) is an inhibitory neurotransmitter that has been shown to have anxiolytic, anti-aging and inhibitory effects on nervous system excitability. However, there are few reports on the prevention and control of the toxicity of nano-metal ions by GABA. In zebrafish, ZnO-NPs exposure led to increased mortality and behavioral abnormalities of larva, which could be moderated by GABA intervention. Similar results were investigated in Caenorhabditis elegans, showing lifespan extension, abnormal locomotor frequency and behavior recovery when worms fed with GABA under ZnO-NPs exposure. Moreover, GABA enhanced antioxidant enzyme activities by upregulating the expression of antioxidant-related genes and thus scavenged excessive O2−. In the case of ZnO-NPs exposure, inhibition of nuclear translocation of DAF-16 and SKN-1 was restored by GABA. Meanwhile, the protective effect of GABA was blocked in daf-16 (−) and skn-1 (−) mutant, suggesting that DAF-16/FoxO and SKN-1/Nrf2 pathways is the key targets of GABA. This study provides a new solution for the application of GABA and mitigation of metal nanoparticle neurotoxicity.

In vitro ovicidal and larvicidal activity of a hydroalcoholic extract and its fractions from Cyrtocarpa procera fruits on Haemonchus contortus

This study describes the in vitro anthelmintic effect of a hydroalcoholic extract (HA-E) and its fractions from Cyrtocarpa procera fruits against Haemonchus contortus eggs and infective larvae. The HA-E was subjected to bipartition using ethyl acetate, which resulted in an aqueous fraction (Aq-F) and an organic fraction (EtOAc-F). The HA-E and both fractions were tested using the egg hatching inhibition assay (EHIA) and the larval mortality test (LMT). Fractionation of the EtOAc-F was achieved using different chromatographic processes, i.e., open glass column and HPLC analysis. Fractionation of the EtOAc-F gave 18 subfractions (C1R1-C1R18), and those that showed the highest yields (C1R15, C1R16, C1R17 and C1R18) were subjected to anthelmintic assays. The HA-E and the EtOAc-F displayed 100% egg hatching inhibition at 3 and 1 mg/mL, respectively, whereas Aq-F exhibited 92.57% EHI at 3 mg/mL. All subfractions tested showed ovicidal effect. Regarding the larval mortality test, HA-E and EtOAc-F exhibited a larvicidal effect higher than 50% at 50 and 30 mg/mL, respectively. The subfractions that showed the highest larval mortality against H. contortus were C1R15 and C1R17, with larval mortalities of 53.57% and 60.23% at 10 mg/mL, respectively. Chemical analysis of these bioactive subfractions (C1R15 and C1R17) revealed the presence of gallic acid, protocatechuic acid, and ellagic acid. This study shows evidence about the ovicidal and larvicidal properties of C. procera fruits that could make these plant products to be considered as a natural potential anthelmintic agents for controlling haemonchosis in goats and sheep.

First report of native Beauveria bassiana and Trichoderma ghanense as bio-control agents against Spodoptera frugiperda in Ghana

Signs of mycosis on mummified Spodoptera frugiperda cadavers were found in maize fields in the Ejura-Sekyedumase and Wa Municipalities in the Ashanti Region and Upper West Region of Ghana, respectively, during the 2019 to 2022 growing seasons. Two major signs were observed on two sets of cadavers that were collected. These were dense white powdery colony encapsulating cadavers, and green powdery growth engulfing cadavers. The causal fungi were confirmed as Beauveria bassiana from white cadaver mummies and Trichoderma ghanense from green cadaver mummies based on morphological and molecular characteristics. The two fungi were assessed for their lethal effect against 3rd instar larvae of S. frugiperda by artificial inoculation. The inoculation was performed with conidia suspension of 1 × 108 conidia/ml for each fungi. B. bassiana caused 55% larval mortality, which was significantly higher (p < 0.05) than the 32.5% caused by T. ghanense. No larval mortality was recorded for the control. Artificially inoculated 3rd instar S. frugriperda larvae exhibited signs similar to those observed on infected larvae collected from the field. The respective pathogens were re-isolated and the morphology matched B. bassiana and T. ghanense, respectively. The findings indicate the potential of the two fungi as bio-control agents for management of S. frugiperda on maize. The obtained result could find application in the formulation of bio-pesticides for S. frugiperda management.

Efficacy of Some Aquatic Plant Extracts on the Khapra Beetle &lt;i&gt;Trogoderma Granarium&lt;/i&gt; Everts

Khapra beetle is one of the dangerous pests that infests fields and warehouses, and using safe alternatives such as plant extracts is one of the options. The efficacy of aqueous extracts of Achillea fragrantissima and Chenopodium album (5, 10, 15%) at different exposure durations (48, 96, and, 144 hr) has been evaluated in this study. Mortality of fifth-instar larvae and adults was observed, and it was shown that an exposure period of 144 hr was superior with hot and coldwater extracts of C. album. The results also showed that maximum mortality of fifth instar larva (65,000 and 92,405%), respectively, was at a concentration of 15% for the cold and boiling water extracts. Mortality of adults was maximum with cold and hot water extracts of the C. album, at an exposure period of 144 hr, as well as at 15% (36,809 and 55,671%, and (51,299 and 74,671%, respectively). Thus the results showed that C. album extract was superior, there existed a significant effect of the interaction between (extract type, concentration, and treatment duration) on the mortality of larvae and adults.

Evaluation of Reproductive and Growth Performance in New Breeds and Hybrids of Bivoltine Silkworm (Bombyx mori L.)

This study aimed to evaluate the reproductive and growth performance of recently developed bivoltine silkworm parental breeds and their hybrids to improve silk production and ensure the long-term sustainability of sericulture. Utilizing five thermotolerant pure breeds (B1, B2, B4, B6 and B8) and four popular CSR breeds (CSR2, CSR27, CSR6 and CSR26), six foundation crosses and eight double hybrids were developed and evaluated for ten traits. Results showed that B1 exhibited superior performance in fecundity (524.00), hatching percentage (98.67%), dead eggs (1.33%), pupal weight (1.31 g), pupation rate (96.84%) and cumulative survival index (CSI) (93.84%). The foundation cross B1 × B2 excelled in fecundity (553.67), larval mortality (2.67%), pupal weight (1.53 g), ERR (97.33%) and CSI (93.27%) while FC2 performed better in hatching percentage (97.60%), dead eggs percentage (2.40%), fifth instar larval duration (157.21 h), fifth instar larval weight (35.64 g/10 larvae) and pupation rate (96.33%). Among the double hybrids, (B1 × B2) × (FC1) demonstrated superior performance in fecundity (614.33), larval mortality (1.41%), pupal weight (2.06 g), ERR (98.59%) and CSI (97.24%). FC2 × FC1 excelled in larval duration (141.11 h), fifth instar larval weight (51.96 g/10 larvae) and pupation rate (98.94%). Double hybrids showed significant improvements compared to parental breeds and foundation crosses, indicating potential for field testing and further development. These results indicate the potential of using the newly identified breeds and foundation crosses in silkworm breeding, particularly in developing new double hybrids as an alternative to established CSR breeds and hybrids.

First Report of &lt;i&gt;Metarhizium Rileyi&lt;/i&gt; Infecting Fall Army Worm &lt;i&gt;Spodoptera Frugiperda&lt;/i&gt; (J E Smith) From Nepal

The fall army worm (FAW) Spodoptera frugiperda (Noctuidae: Lepidoptera) is an invasive polyphagous pest that prefers maize crop in the newly invaded regions like Africa, Asia, Australasia, and some Pacific Islands and was recorded in Nepal in the year 2019. Surveys were conducted in the maize fields to collect various natural enemies for utilization in the management of FAW. Several larvae infected with an unidentified fungus were collected and subjected to morphological and molecular analysis for identification as Metarhizium rileyi. In bioassay studies we observed percent larval mortality at 1x108 conidia/ml suspension. This is the first report of M. rileyi infecting S. frugiperda from Nepal.

Mealworm (Tenebrio molitor) rearing and growth optimization as a sustainable food source using various larval diets under laboratory conditions

AbstractA new approach to supplying foods and feeds that are high in protein has emerged in the form of insect farming. Insect‐farmed food may also serve as part of a sustainable diet for humans. The mealworm Tenebrio molitor L. (Coleoptera: Tenebrionidae) is the most widely consumed insect worldwide. In this study, 10 larval diets were tested in the laboratory to determine their suitability for mass‐feeding mealworms at 28 °C and 60–65% r.h., in a completely randomized design. Various larval diets had a significant impact on the biological features of T. molitor. The larvae, pupae, and adults were greater on diets containing wheat bran alone and wheat bran in combination with maize and porridge. Larval mortality was significantly higher (60.7%) on a diet with porridge alone, compared to a control diet, followed by diets with porridge in combination with barley (55.6%) or maize (50.6%). Sex ratio was also affected by the tested larval diets. Female hatch (65%) was higher than male hatch (35%) on diet containing wheat bran alone as compared to all other diets. The findings contribute to optimizing mealworm rearing practices for sustainable food production and promote the potential use of mealworms as a viable protein source – especially livestock diets based on mealworm reared on wheat bran show promise.

Phytochemicals, antioxidant ability and in vitro anthelmintic activity of crude extracts from Vitexnegundo leaves against Haemonchus contortus.

The aim of this study was to evaluate the in vitro anthelmintic effect of crude aqueous, methanol, ethanol, hydro alcohol and acetone extracts of Vitex negundo leaves against Haemonchus contortus eggs and larvae. Phytochemical analysis to identify the number of compounds in extracts was done by chemical tests and gas chromatography coupled to a mass spectrophotometer detector (GC-MS). First off all the effectiveness of dried plant materials was evaluated on larval development by mixing powdered material (no nano particles) to faecal cultures from donor sheep. Adding powder to the faecal culture resulted into 100% inhibition in larval development at 200 and 300mg/g of faeces. The anthelmintic activity was assessed using the egg hatch assay (EHA) and the larval mortality assay (LMA). Comparison of mean inhibition percentage of egg embryonation, mean inhibition percentage of egg hatching and mean percentage of larval mortality at different concentrations with control was performed by one-way ANOVA. The means were compared for statistical significance using DMRT at P < 0.05. For both the assays, 50% inhibitory concentration (IC50) and lethal concentration (LC50) were calculated by probit analysis. Chemical test revealed presence of high concentration of saponin and flavoinoids and moderate concentration of total phenols in leaves. The antioxidant activity (radical scavenging activity, RSA %) measured was 35.47%. On GC-MS, the methanolic leaves extract revealed 30 phyto-compounds. On EHA, there was marked effect on inhibition of egg hatching by aqueous, hydro alcohol and acetone extracts. On LMA all the five extracts showed excellent larvicidal activity. V. negundo leaves methanol extract mediated silver nanoparticles were found very effective at much lower concentrations as compared to crude methanol extract. The results indicated that the V. negundo leaves crude extracts possessed excellent in vitro ovicidal and larvicidal properties against H. contortus which needs more investigation, especially in vivo trials for the control of parasite.

Virulence phenotypes differ between toxigenic Vibrio parahaemolyticus isolated from western coasts of Europe

Vibrio parahaemolyticus is the leading bacterial cause of gastroenteritis associated with seafood consumption worldwide. Not all members of the species are thought to be pathogenic, thus identification of virulent organisms is essential to protect public health and the seafood industry. Correlations of human disease and known genetic markers (e.g. thermostable direct hemolysin (TDH), TDH-related hemolysin (TRH)) appear complex. Some isolates recovered from patients lack these factors, while their presence has become increasingly noted in isolates recovered from the environment. Here, we used whole-genome sequencing in combination with mammalian and insect models of infection to assess the pathogenic potential of V. parahaemolyticus isolated from European Atlantic shellfish production areas. We found environmental V. parahaemolyticus isolates harboured multiple virulence-associated genes, including TDH and/or TRH. However, carriage of these factors did not necessarily reflect virulence in the mammalian intestine, as an isolate containing TDH and the genes coding for a type 3 secretion system (T3SS) 2α virulence determinant, appeared avirulent. Moreover, environmental V. parahaemolyticus lacking TDH or TRH could be assigned to groups causing low and high levels of mortality in insect larvae, with experiments using defined bacterial mutants showing that a functional T3SS1 contributed to larval death. When taken together, our findings highlight the genetic diversity of V. parahaemolyticus isolates found in the environment, their potential to cause disease and the need for a more systematic evaluation of virulence in diverse V. parahaemolyticus to allow better genetic markers.

Keap1 Negatively Regulates Transcription of Three Counter-Defense Genes and Susceptibility to Plant Toxin Gossypol in Helicoverpa armigera.

Expressions of a wide range of cytoprotective counter-defense genes are mainly regulated by the Keap1-Nrf2-ARE signaling pathway in response to oxidative stress from xenobiotics. Gossypol is the major antiherbivore secondary metabolite of cotton, but how the polyphagous pest Helicoverpa armigera copes with this phytochemical to utilize its favorite host plant cotton remains largely elusive. In this study, we first suppressed the Keap1 gene in newly hatched larvae of cotton bollworm by feeding them the siRNA diet for 4 days. All of the larvae were subsequently fed the artificial diet supplied with gossypol or the control diet for 5 days. We identified that the knockdown of the Keap1 gene significantly decreased larval mortality and significantly increased the percentages of larval survival, reaching the fourth instar, compared with ncsiRNA when exposed to a diet containing gossypol. Three counter-defense genes CYP9A17, CYP4L11 and UGT41B3, which were related to the induction or metabolism of gossypol according to the report before, were all significantly up-regulated after the knockdown of the Keap1 gene. The Antioxidant Response Elements (AREs) were also detected in the promoter regions of the three counter-defense genes above. These data indicate that the suppression of the Keap1 gene activates the Keap1-Nrf2-ARE signaling pathway, up-regulates the expressions of counter-defense genes involved in the resistance of oxidative stress and finally contributes to reducing the susceptibility of gossypol. Our results provide more knowledge about the transcriptional regulation mechanisms of counter-defense genes that enable the cotton bollworm to adapt to the diversity of host plants including cotton.

Effect of Lupinus (Fabaceae) on the biology of Spodoptera frugiperda (Lepidoptera, Noctuidae)

The activity of aqueous extracts at 5% from seeds of six wild lupin species wasevaluated to determine the effect on the growth, development and viability ofSpodoptera frugiperda. Six grams of powdered seeds of each plant sample wereadded in 120 mL of water during 24 h; then this aqueous extract was used in theexperiment. The diet was spilled in glass vials after neonates larvae of S.frugiperda were put in to vials. Immediately they were kept in the raise room at 27± 0.5°C, 60 ± 10% relative humidity and 14 h light, until larvae changed to pupaephase. Length and mortality of larvae and pupae phases, and weight pupae in eachdiet, were recorded. The results showed that at the evaluated concentration, theactivity of the extract was insectistatic; inhibiting feeding and growth of S.frugiperda. The extract was not insecticidal.

Exposure of Beauveria bassiana from different sites to changes in behaviour and mortality of oil palm beetle larvae: Oryctes rhinoceros

The potential of Beauveria bassiana as a biological control agent for Oryctes rhinoceros is still rarely reported. The objective of the research was to analyse the behaviour and mortality of O. rhinoceros larva due to exposure to several of B. bassiana. The research method used the Completely Randomized Design and analysis with 5% Tukey’s test. The results showed that B. bassiana form different sites had the potential to be developed as an effective biological control product in controlling O. rhinoceros as a part of novelty to explore the biodiversity of native biological control in Indonesia. Larvae of O. rhinoceros that were exposed to and infected with B. bassiana experienced changes in movement activity and decreased appetite as well as differences in cuticle colour, the initially limp body will turn stiff while the cuticle colour will change and the next symptom will be white mycelium growth which was proven by the SEM. The B. bassiana from Jember 1 had the highest mortality rate of 37.5% at 30 days after application even though it had the lowest spore density. These results indicate something new and require further study regarding the toxin content of each B. bassiana from different sites.

Twinstar is a chitin synthase interacting protein with an essential role in insect cuticle biosynthesis

Chitin is an abundant natural biopolymer that plays a crucial role in insect growth and development as a fundamental structural component of the exoskeleton. The membrane-integral β-glycosyltransferase, chitin synthase, has been identified as the central component in chitin biosynthesis. However, the precise roles of other proteins in facilitating chitin synthase in chitin biosynthesis remain unclear. In this study, we employed split-ubiquitin membrane yeast two-hybrid (MYTH) and pull-down assays to demonstrate the physical interaction between Twinstar (Tsr), a small molecular protein in the actin-depolymerizing factor ADF/Cofilin protein family, and chitin synthase Krotzkopf verkehrt (Kkv) in Drosophila melanogaster in vitro. The RNA interference (RNAi)-mediated global knockdown of Tsr in D. melanogaster resulted in larval lethality. Furthermore, targeted suppression of Tsr in the tracheal and epidermal tissues also led to larval mortality, while knocking down Tsr in the wing tissues led to wrinkled wings. Additionally, silencing Tsr not only reduced the chitin content in the first longitudinal vein of the wings but also led to the absence of the chitin lamellar structure. To validate the functional conservation of Tsr in other insect orders, the two agricultural pests Ostrinia furnacalis and Tribolium castaneum, representing lepidoptera and coleoptera insects, respectively, were investigated. Knockdown experiments targeting the Drosophila Tsr orthologues OfTsr in O. furnacalis and TcTsr in T. castaneum produced abnormal larvae during molting or pupation in O. furnacalis and lethality in T. castaneum. Our findings not only improve our knowledge of the chitin biosynthesis machinery in insect cuticles but also provide new potential targets for the control of major agricultural pests.