1st Instar Larvae Research Articles

Synthesis, characterization and toxicity assessment of chlorantraniliprole nanoemulsion against Helicoverpa armigera

Helicoverpa armigera is a highly devastating and polyphagous pest attacking on different field crops. Effective management of this pest is essential for sustainable agriculture. While traditional chemical pesticides are widely used for their immediate results and ease of application, nanoemulsions offer a promising alternative, paving the way for advanced, efficient pesticide formulations.The current research aimed to develop oil-in-water (O/W) chlorantraniliprole stable nanoemulsion using combinations of different solvents (butanol, dimethyl sulfoxide and toluene), tween- 80 (non-ionic surfactant) and sodium laurel sulfonate (anionic co-surfactant) to reduce the dose and increase toxicity of chlorantraniliprole as targeted pesticide. Ultrasonication, a high energy emulsification method, is adopted to obtain the particle size in nanometre (nm). The nano size of emulsion is confirmed by Dynamic light scattering (62.43 nm), Scanning electron microscopy (55 ± 5 nm), X-ray diffraction (42.9 nm), and Transmission electron microscopy (22.5 ± 2.5 nm), Stability of synthesized nanoemulsion is confirmed via, centrifugation, freeze thaw cycle, heating cooling test, and zeta potential. Moreover, the toxicity of chlorantraniliprole nanoformulations is evaluated against H. armigera and compared with commercially available chlorantraniliprole formulation (18.5 SC). Chlorantraniliprole nanoemulsion formulation is 3.3 times more toxic than commercial formulation using topical application method (3rd instar larvae) and 2.2 times more toxic than commercial formulation using diet incorporation method (1st instar larvae). Consequently, this cutting-edge research holds great potential for revolutionizing targeted pesticide delivery systems, paving the way for more effective and precise pest management in agriculture.

Lifecycle Completion and Reproductive Improvement of Chrysoperla carnea (Stephens) (Neuroptera: Chrysopidae), Following a Prey Shift Routine During Larval Development.

The present study aimed to assess the lifecycle completion and reproductive improvement of green lacewing, Chrysoperla carnea (Stephens), larval instars in a prey shift routine while alternatively feeding on motiles of two-spotted spider mite, Tetranychus urticae Koch (natural prey), and the eggs of almond moth, Ephestia cautella (Walker) (factitious prey). The results showed that C. carnea larvae successfully completed their lifecycle and life span in the treatments where either the 2nd (T2) or 3rd (T3) instar larvae were fed with the eggs of E. cautella while other instars were provided with motiles of T. urticae, i.e., 1st-3rd and 1st-2nd, respectively. The predator larvae failed to transform from pupae to adults when only the 1st instar larvae were fed on the E. cautella eggs (T1). Similar results were found when all predator instars were reared on the motiles of T. urticae (T4). The shortest mean total larval duration and preoviposition period, the highest fecundity and fertility, and balanced sex ratio were recorded when 3rd instar larvae were fed only on E. cautella eggs (T3). The longest adult longevity and oviposition period were recorded when only the 2nd instar larvae were fed on E. cautella eggs (T2). The findings of the present study would be helpful in developing a self-sustained mite biological control strategy using C. carnea and provide a roadmap for future studies to elucidate the larval release efficiency of C. carnea with the supplementary diet for the management of greenhouse mite pests.

Comparative analysis of the insecticidal activity of Biogenic Zinc Oxide Nanoparticles, Iron Oxide Nanoparticles and piper nigrum fruits extract against 1st instar larva of House fly, (Musca domestica) (Diptera: Muscidae)

Comparative analysis of the insecticidal activity of Biogenic Zinc Oxide Nanoparticles, Iron Oxide Nanoparticles and piper nigrum fruits extract against 1st instar larva of House fly, (Musca domestica) (Diptera: Muscidae)

Isolation and characterization of halophilic Bacillus thuringiensis from local Egyptian sites and their potential against Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)

BackgroundBacillus thuringiensis (Bt) serves as a peculiar soil microbe that is Gram-positive, rod-shaped, spore-forming, and aerobic facultative. It endures across a range of conditions and has been demonstrated to be an effective biological pest control agent against several insect pests. The main objective of the study is to isolate and recognize Bt bacteria from saline environments in Egyptian soil for biological surveillance uses.ResultsIsolates three, four, five, ten, eleven, twelve, and thirteen were found to be moderately tolerant, while isolate eight showed highly tolerant, but isolate six was found to be borderline extreme halophile. On the other hand, the other isolates showed extreme halophiles. To find out the presence of crystals, a scanning electron microscope was carried out and the result showed that the crystals ranged from spherical to bi-pyramidal spherical. Molecular analysis was carried out using universal primers to confirm the presence of cry genes. The results showed that 84.6% of the isolates contain cry seven and eight, whereas cry one and four are distributed by 92.3%. Cry two genes were found to be 100 percent in all tested and isolated cultures. Cry genes were present and distribution among isolated was detected using gene-specific primers. cry3Ba1 and cry8B showed the lowest and equal distribution among the isolates, while the cry2Ab2gene frequency distribution was (84.6%) among isolated cultures. Around six cry genes were found to be absent in all isolated cultures. The bioinsecticidal activity and bioassays were carried out to check the potential effect of halophilic Bt isolates against 1st instar larvae of the Fall Armyworm (FAW) Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae). Mortality was calculated after infecting S. frugiperda with thirteen Bt isolates. The most virulent Bt isolate was isolate 7, followed by isolate 10, with LC50 values of 9.542 × 107 spores/ml and 1.289 × 108 spores/ml, respectively, after 72 h. Morphological traits of the larvae were carried out, to check the effect of Bt isolates as a biological control against the FAW.ConclusionTo protect and enhance the sustainability of field crops and vegetables, especially in new high salinity lands, from disease-causing insects such as FAW, it is necessary to identify new strains of Bt from these saline lands. In this field, local isolates of Bt from several regions in Egypt known for their high levels of salinity were documented and showed an effective bioinsecticide on FAW larvae.

Isolation and Evaluation of Indigenous Isolates of Beauveria bassiana and Synergistic Control of Spodoptera frugiperda with the Parasitoid Microplitis prodeniae.

Entomopathogenic fungi (EPFs) are capable of infecting a variety of insect pests and are widely used as biopesticides worldwide. This study intended to isolate indigenous EPFs from cadavers of Protaetia brevitarsis and investigate their effects on the fall armyworm Spodoptera frugiperda (FAW), a globally widespread invasive pest. Morphological and molecular analyses confirmed four native EPF strains all belong to Beauveria bassiana. Pathogenicity tests showed they were virulent toward FAW 1st instar larvae. The application of EPFs either by dipping or spraying significantly increased the larval mortalities compared to the control group, with corrected mortalities ranging from 92% to 73% after dipping in a fungal suspension of 108 conidia/mL, and those ranging from 76% to 35% after spraying with a fungal suspension of 107 conidia/mL. Our findings revealed the infectivity of four strains to FAW larvae significantly changed in a dose- and time-dependent manner. In addition, the combination use of the local B. bassiana strain and parasitoid Microplitis prodeniae resulted in a significantly enhanced S. frugiperda 3rd instar larval mortality compared to a single inoculation with one of them, suggesting an apparent synergistic effect for the joint application of these two biological control agents. The mortality inflicted by B. bassiana was probably promoted by the release of parasitoids, since the parasitoids' movements and attacks could strengthen the fungal distribution and infection processes. This study underscores the potential of the combination use of EPFs and parasitoids against S. frugiperda early-instar larvae, and provides insights into the consequences of integrating these EPFs into integrated pest management systems.

Exploring the ameliorative potential of rutin against High-Sucrose Diet-induced oxidative stress and reproductive toxicity in Drosophila melanogaster

Sucrose is a vital ingredient in numerous food items consumed regularly. However, exposure to excessive sucrose for a prolonged period can promote health issues. The reproductive system has a delicate physiology that can be targeted by various chemical stressors, including sucrose. Hence, the present in vivo study aims to unveil the impacts of High-Sucrose Diet (HSD) on the reproductive fitness of Drosophila melanogaster. In addition, the present work has also assessed the protective potential of a bioactive compound, rutin, against it. Here, first instar larvae were exposed to HSD (30 %) alone and in combination with rutin (100–300 µM) till their adult stage. HSD disturbed sex comb morphology in adult males, while fecundity and hatchability of eggs in females. Moreover, HSD triggered gonadal ROS production, oxidative stress, and modulated endogenous antioxidants such as SOD, catalase, and glutathione in both sexes. Nuclear fragmentation and tissue injuries, along with protein and lipid oxidation, were also apparent. Elevated levels of cytosolic Iron suggested an active Fenton reaction in adults. Further, HSD modulated the activities of reproductive and metabolic mediators, including vitellogenin, malate dehydrogenase, glucose-6-phosphate dehydrogenase, and angiotensin-converting enzymes that are critical to maintain the overall reproductive fitness. Interestingly, co-treatment with rutin, mainly at 200 µM, mitigated these adverse effects and restored reproductive fitness. The protective potential of rutin might be attributed to its ability to normalize redox homeostasis, reduce oxidative stress, and optimize critical enzymes involved in reproductive physiology. These findings suggest that rutin has potential therapeutic implications for counteracting the reproductive hazards induced by HSD.

Studies On The Statistical Analysis And Impact Of Seasonal Temperature And Relative Humidity Variation On The Development Of A Flesh Fly Sarcophagadux(Thomson 1869)(Diptera Sarcophagidae)

Background: The fleshfly Sarcophaga dux (Diptera;Sarcophagidae) is found in Indian Sub continent is of limited forensic significance as its poorly known larval development relationship with temperature and relative humidity. Objective: The objective of this study was to determine the effect of the temperature and relative humidity in the larval development of this fly in two seasons, Summer and Rain, so that the results of this study may enabling the forensic use of this fly in the estimation of Post Mortem Interval(PMI). Material and Method: The present study was carried out in the outer premises of the Regional Forensic Science Laboratory Building, Jagdalpur, Bastardist,Chhattishgarh, India. The fresh meat of male goat was used for rearing of larvae. The effect of temperature and relative humidity on the larval developmental time of this fly in two seasons, Rainy and Summer were studied in normal condition. Statistical analysis of variance was performed and standard significance level of P<0.05 was taken as standard. Results: The results of this study shown that in Rainy season it was completed in 264 h (11 days) when the average temperature ranged from 21.8°C to 24.5°C and the average humidity ranged from 79.5% to 95.5% where as in Summer it was completed in 528 hrs (22 days), when the temperature ranged from 26.5°C to 35.1°C and the humidity ranged from 34% to 67.5%. The P value was found to be P<<.05 in Temperature variation; andP<<0.05 in Humidity variation in different stages(1st instar larvae, 2nd instar larvae, 3rd instar larvae, Prepupae, Pupae and Adult). Conclusion: In the present study we have found that fluctuating temperature and relative humidity of outer condition in two seasons, Rainy and Summer effect the life cycle duration of the fly from the larvi position up to adult eclosion and statistical analysis has proved it to significant rejecting null hypothesis. This data can be further used in the estimation of Post Mortem Interval (PMI) in Homicide and Suspicious cases of both human and wild life poaching death cases.

Resistance Realized Heritability and Fitness Cost of Cyproflanilide in Rice Stem Borer, Chilo suppressalis (Lepidoptera: Pyralidae)

The rice stem borer (RSB) Chilo suppressalis is a devastating rice pest with resistance to a number of insecticides. Recently, the new meta-diamide insecticide cyproflanilide has been considered an effective insecticide to control RSB. However, its resistance risk has not been reported. In the present study, we aimed to assess the resistance risk and evaluate the fitness cost after the RSB was exposed to cyproflanilide. After five generations of selection, the resistance level of RSB increased by 1.5-fold. When h2 was 0.125, a 10-fold resistance increase in the LD50 values was expected in fourteen and thirty-one generations at the selection intensity of 90% and 50%, respectively. The selected population (RSB-SEL) had significant differences in the developmental duration of eggs, 1st, 2nd, 3rd, and 6th instar larvae, and female pupae compared to the unselected population (RSB-UNSEL). Besides, the adult longevity was shortened, and the average pupal weight of males, the emergence rate, the sex ratio, the oviposition, the mean fecundity, and the full life cycle rate were decreased in RSB-SEL. The intrinsic rate of increase (r), the net reproductive rate (R0), and the finite rate of increase (λ) of RSB-SEL were significantly lower than those of RSB-UNSEL, while the mean generation time (T) of RSB-SEL was significantly longer than that of RSB-UNSEL. Based on the results of the prediction of the generations required for a 10-fold resistance increase in the LD50, a potential risk of resistance development exists in RSB after continuous and excessive use of cyproflanilide. These results will be useful in designing the dose of cyproflanilide to control C. suppressalis in field.

Biological and Physiological Changes in Spodoptera frugiperda Larvae Induced by Non-Consumptive Effects of the Predator Harmonia axyridis

The effects of predatory natural enemies on their prey or hosts involve both consumption and non-consumptive effects. This study investigated the non-consumptive effects of the predator, Harmonia axyridis (Coleoptera: Coccinellidae) on 1st, 2nd and 3rd instar larvae of Spodoptera frugiperda. We exposed larvae of different instars to the predator and assessed various parameters using a combination of biological and biochemical methods. Exposure to the predator significantly affected the growth and development of the S. frugiperda caterpillars. Firstly, the developmental duration of S. frugiperda larvae in the 1st–3rd instars and the pupal stage were notably prolonged. Moreover, we observed significant effects on pupal mass, pupal abnormality rate and emergence rate. These non-consumptive effects were gradually weakened with an increase in the larval stage exposed to the predator. Antioxidant enzyme activities including catalase (CAT) peroxidase (POD) and superoxide dismutase (SOD) activity increased significantly. Additionally, organismal triglyceride, trehalose and glycogen content were significantly altered by non-consumptive effects, while protein content showed no significant change. Spodoptera frugiperda larvae increased the activity of antioxidant enzymes in response to potential predators to mitigate oxidative stress and reduce cellular and tissue damage. This resources redistribution towards survival may inhibit growth and development of the species and further exacerbate these non-consumptive effects. These findings highlight the importance of considering non-consumptive effects in pest-management strategies to optimize control measures in agricultural systems.

Effects of developmental stages, sex difference, and diet types of the host marmalade hoverfly (Episyrphus balteatus) on symbiotic bacteria.

Symbiotic bacteria play key roles in a variety of important life processes of insects such as development, reproduction and environmental adaptation, and the elucidation of symbiont population structure and dynamics is crucial for revealing the underlying regulatory mechanisms. The marmalade hoverfly (Episyrphus balteatus) is not only a remarkable aphid predator, but also a worldwide pollinator second to honeybees. However, its symbiont composition and dynamics remain unclear. Herein, we investigate the symbiotic bacterial dynamics in marmalade hoverfly throughout whole life cycle, across two sexes, and in its prey Megoura crassicauda by 16S rRNA sequencing. In general, the dominant phyla were Proteobacteria and Firmicutes, and the dominant genera were Serratia and Wolbachia. Serratia mainly existed in the larval stage of hoverfly with the highest relative abundance of 86.24% in the 1st instar larvae. Wolbachia was found in adults and eggs with the highest relative abundance of 62.80% in eggs. Significant difference in species diversity was observed between the adults feeding on pollen and larvae feeding on M. crassicauda, in which the dominant symbiotic bacteria were Asaia and Serratia, respectively. However, between two sexes, the symbionts exhibited high similarity in species composition. In addition, our results suggested that E. balteatus obtainded Serratia mainly through horizontal transmission by feeding on prey aphids, whereas it acquired Wolbachia mainly through intergeneration vertical transmission. Taken together, our study revealed the effects of development stages, diet types and genders of E. balteatus on symbionts, and explored transmission modes of dominant bacteria Serratia and Wolbachia. Our findings lay a foundation for further studying the roles of symbiotic bacteria in E. balteatus life cycle, which will benefit for revealing the co-adaptation mechanisms of insects and symbiotic bacteria.

Temperature-dependent development and population growth of the invasive pest Cassida vittata vill. In sugar beet crops

The sugar beet flea beetle, Cassida vittata Vill. is an invasive pest that significantly damages sugar beet crops in Morocco by affecting the leaves. To gain a deeper understanding of how temperature influences the development and reproduction of C. vittata and to facilitate its population management, laboratory-based experiments were conducted. Individuals were subjected to a range of constant temperatures spanning from 15 °C to 36 °C. The most extended lifespans for both adult females and males were recorded at 30 °C, with respective longevities of 62.8 and 62.5 days. The highest likelihood of eggs maturing into the adult stage occurred at 25 °C, with survival rates of 39.0% for females and 37.5% for males. Preadult survival rates displayed significant variability with temperature, with the highest survivorship observed at 25 °C (76.5%). The maximum average number of offspring produced per female reached 225.1 eggs at 25 °C. Daily reproduction, maximum total fecundity, and maximum daily fecundity were also notably higher at 25 °C. The population growth parameters exhibited their most favorable values at 25 °C, with r (0.0933 d−1), and R0 (87.15 offspring per female) being highest at this temperature. The temperature thresholds for the complete pre-adult stage (1st instar larva to adult) were estimated to be 9.4 °C for females and 9.0 °C for males. The thermal constants from the 1st instar larva to the adult stage were calculated to be 526.3 and 476.2 DD, respectively. These findings indicate that the most favorable temperature range for the population growth of this invasive pest falls between 25 °C and 30 °C.

RNAi-mediated pest control targeting the Troponin I (wupA) gene in sweet potato weevil, Cylas formicarius.

The sweet potato weevil (Cylas formicarius) is a critical pest producing enormous global losses in sweet potato crops. Traditional pest management approaches for sweet potato weevil, primarily using chemical pesticides, causes pollution, food safety issues, and harming natural enemies. While RNA interference (RNAi) is a promising environmentally friendly approach to pest control, its efficacy in controlling the sweet potato weevil has not been extensively studied. In this study, we selected a potential target for controlling C. formicarius, the Troponin I gene (wupA), which is essential for musculature composition and crucial for fundamental life activities. We determined that wupA is abundantly expressed throughout all developmental stages of the sweet potato weevil. We evaluated the efficiency of double-stranded RNAs in silencing the wupA gene via microinjection and oral feeding of sweet potato weevil larvae at different ages. Our findings demonstrate that both approaches significantly reduced the expression of wupA and produced high mortality. Moreover, the 1st instar larvae administered dswupA exhibited significant growth inhibition. We assessed the toxicity of dswupA on the no-target insect silkworm and assessed its safety. Our study indicates that wupA knockdown can inhibit the growth and development of C. formicarius and offer a potential target gene for environmentally friendly control.

Orthotospovirus iridimaculaflavi (iris yellow spot virus): An emerging threat to onion cultivation and its transmission by Thrips tabaci in India

The yellow spot disease caused by the virus species Orthotospovirus iridimaculaflavi (Iris yellow spot virus-IYSV), belonging to the genus Orthotospovirus, the family Tospoviridae, order Bunyavirales and transmitted by Thrips tabaci Lindeman. At present, emerging as a major threat in onion (Allium cepa) in Tamil Nadu, India. The yellow spot disease incidence was found to be 53–73 % in six districts out of eight major onion-growing districts surveyed in Tamil Nadu during 2021–2023. Among the onion cultivars surveyed, the cultivar CO 5 was the most susceptible to IYSV. The population of thrips was nearly 5–9/plant during vegetative and flowering stages. The thrips infestation was 34–60 %. The tospovirus involved was confirmed as IYSV through DAS-ELISA, followed by molecular confirmation through RT-PCR using the nucleocapsid (N) gene. The predominant thrips species present in onion crops throughout the growing seasons was confirmed as Thrips tabaci based on the nucleotide sequence of the MtCOI gene. The mechanical inoculation of IYSV in different hosts viz., Vigna unguiculata, Gomphrena globosa, Chenopodium amaranticolor, Chenopodium quinoa and Nicotiana benthamiana resulted in chlorotic and necrotic lesion symptoms. The electron microscopic studies with partially purified sap from onion lesions revealed the presence of spherical to pleomorphic particles measuring 100–230 nm diameter. The transmission of IYSV was successful with viruliferous adult Thrips tabaci in cowpea (Cv. CO7), which matured from 1st instar larva fed on infected cowpea leaves (24 h AAP). Small brown necrotic symptoms were produced on inoculated plants after an interval of four weeks. The settling preference of non-viruliferous and viruliferous T. tabaci towards healthy and infected onion leaves resulted in the increased preference of non-viruliferous thrips towards infected (onion-61.33 % and viruliferous thrips towards healthy onion leaves (75.33 %). The study isolates shared 99–100 % identity at a nucleotide and amino acid level with Indian isolates of IYSV in the N gene. The multiple alignment of the amino acid sequence of the N gene of IYSV isolates collected from different locations and IYSV isolates from the database revealed amino acid substitution in the isolate ITPR4. All the IYSV isolates from India exhibited characteristic amino acid substitution of serine at the 6th position in the place of threonine in the isolates from Australia, Japan and USA. The phylogenetic analysis revealed the monophyletic origin of the IYSV isolates in India.

Death Feigning in Larvae of Scorpionflies (Mecoptera: Panorpidae): Frequency and Postural Changes Based on Larval Instars

AbstractDeath feigning is thought to have evolved primarily as a predator avoidance behavior, and has been reported in 10 of the 31 orders of insects. However, there have been no reports of death-feigning behavior in Mecoptera species. We found that larvae of two scorpionfly species, Panorpa japonica and P. pryeri, showed death feigning in response to external stimuli by brush poking stimulation. First, we examined the frequencies of death-feigning postures. The two species showed two different postures of death feigning, “straight” and “ball.” Most of the 1st instar larvae of P. japonica and P. pryeri adopted the straight death-feigning posture. Next, we examined duration of death feigning. As the larval instar progressed, the death-feigning posture shifted from straight to ball in both Panorpa species. In P. japonica, the longest durations of death feigning were found in the 2nd to 3rd instars, while the longest duration of death feigning was found in the late 4th instar in P. pryeri larvae.

Age-Stage, Two-Sex Life Table of an Alarming Quarantine Pest Trogoderma granarium on Stored Products

Trogoderma granarium (Coleoptera: Dermestidae) is a polyphagous pest of stored products and an alarming threat to food security around the globe. The current study is designed to construct the life table of T. granarium on four stored products, i.e., wheat, barley, rice, and maize at controlled conditions (33±1°C, 65% RH, and a photoperiod of 14 : 10 (L:D) h). The incubation period of T. granarium was longer (5.13 d) on maize and shorter (4.30 d) on wheat. The developmental period of 1st, 2nd, 3rd, and 4th larval instars of T. granarium was longer on maize than the other three hosts were significant. Statistical difference between the longevity of females and males was recorded, while no statistically significant difference was found in the preoviposition period. The mean fecundity values observed on wheat, barley, rice, and maize were 31.33±1.62, 26.21±1.49, 28.00±1.35, and 25.58±1.13, respectively. The highest fertility was observed in wheat and the lowest in maize. The net reproductive rate (R0) of T. granarium ranged from 8.66±2.86, 13.16±2.65, 12.53±2.27, and 10.23±2.32 offsprings on wheat, barley, rice, and maize, respectively. The highest gross reproductive value (GRR) was for T. granarium reared on barley (18.31±4.10/d), and the lowest was for maize (15.35±2.95/d). To complete the development, T. granarium took 48.97±0.68 d on maize, significantly higher than on other treatments (p<0.05). The mean generation time (T) of T. granarium was considerably affected by tested products. Based on the results of the present study, maize is a less favorable diet for the development and reproduction of T. granarium and may be considered for the management of this detrimental pest.

Mosquito larvicidal bioassay with the fruit extracts of Diospyros kaki on filarial vector Culex quinquefasciatus

Due to the emergence of resistance of the commonly used synthetic pesticide, vector control is in grave danger. Application of biocontrol agents may be a suitable alternative approach in vector control. The purpose of this study was to evaluate the larvicidal effectiveness of crude and methanolic fruit extracts of Diospyros kaki against Culex quinquefasciatus, the carrier of human filarial worm. The methanol extract showed promising result in biocontrol assay than crude extract. The methanolic extract had the LC50 and LC90 values of 46 ppm and 186 ppm, respectively against 1st instar larvae after 72 hours of exposure, The rate of mortality was positively associated with the solvent extracts in increasing concentrations, having a regression coefficient value near to 1 in each case, showing a definite positive dose-dependent mortality. During chemical characterization of the solvent extract by GC-MS and FT-IR studies, some bioactive compounds like 5,8,11,14,17-Eicosapentaenoic Acid, Methyl 4,7,10,13,16,19-Docosahexaenoate, cyclotrisiloxane hexamethyl and Arsenous acid, tris(trimethylsilyl) ester were identified that showed the larvicidal activity against mosquito larvae either singly or jointly. Thus, D. kaki fruit extract showed excellent potential as a bio control agent for Cx. quinquefasciatus and the isolated bioactive phytochemical may be utilized as a source of a powerful insecticide.

Chronic sub-lethal exposure to clothianidin triggers organismal and sub-organismal-level health hazards in a non-target organism, Drosophila melanogaster

Neonicotinoids are among the most widely used systemic pesticides across the world. These chemicals have gathered significant attention for their potential adverse impacts on non-target organisms. Clothianidin is a novel neonicotinoid pesticide, employed globally to control sucking and chewing types of pests. In nature, various non-target organisms can be exposed to this chemical through contaminated food, water, and air. Nonetheless, extensive investigations demonstrating the sub-lethal impacts of clothianidin on non-target entities are limited. Hence, the present study was aimed to unravel the chronic sub-lethal impacts (LC50 0.74 μg/mL) of clothianidin on a non-target organism, Drosophila melanogaster. The study parameters involved multiple tiers of life ranging from organismal level to the sub-cellular level. 1st instar larvae were exposed to the six sub-lethal concentrations viz. 0.05, 0.06, 0.07, 0.08, 0.09, and 0.1 μg/mL of clothianidin till their 3rd larval instar. Investigations involving organismal level have revealed clothianidin-induced significant reduction in the developmental duration, life span, phototaxis, and physical activities of the treated individuals. Interestingly, the tested compound has also altered the compound eye morphology of treated flies. Study was extended to the tissue and cellular levels where reduced cell viability in gut, brain, and fat body was apparent. Additionally, increased ROS production, nuclear disorganization, and higher lipid deposition were evident in gut of exposed individuals. Study was further extended to the sub-cellular level where chronic exposure to clothianidin up-regulated the major oxidative stress markers such as lipid peroxidation, protein carbonylation, HSP-70, SOD, catalase, GSH, and thioredoxin reductase. Furthermore, the activities of detoxifying enzymes such as CYP4501A1 and GST were also altered. Chronic exposure to clothianidin also triggered DNA fragmentation in treated larvae. In essence, results of this multi-level study depict the ROS-mediated toxicity of clothianidin on a non-target organism, D. melanogaster.

Characterization of the serosal cells surrounding Cotesia kariyai larvae and their role in host immunosuppression

About half of the serosal cells (Scs) of Cotesia kariyai (Ck) eggs are released as teratocytes into the host body cavity after hatching, and another half remain attached to the larval epidermis until the 1st instar larvae of Ck ecdysis to 2nd instars. To investigate the role of the serosal cells surrounding Ck 1st instar larvae (1st Scs) in immune avoidance, Ck 1st instar larvae with and without Scs removed using dispase were transplanted into Mythimna separata larvae (MsLrv), respectively. As a result, Ck 1st instar larvae surrounded by Scs were less susceptible to the MsL encapsulation than Ck 1st instar larvae without the Scs, suggesting that the Scs are involved in suppressing the encapsulation of the MsL. Furthermore, when the granular cells and plasmatocytes of the MsL involved in the encapsulation were incubated in a medium supplemented with proteins extracted from 1st Scs, the plasmatocytes failed to adhere to glass slides, and did not spread their filopodium and lamellipodium. These findings suggest that 1st Scs express proteins that inhibit filopodium and lamellipodium spreading to prevent the MsL plasmatocytes from adhering to Ck larvae.

Effects of Diet on the Gut Bacterial Community of Aldrichina grahami (Diptera: Calliphoridae) across Developmental Stages.

The blowfly, Aldrichina grahami (Diptera: Calliphoridae), is a well-known forensically important insect. Basic data related to A. grahami have been well documented; but despite the pivotal role of gut microbes in various facets of insect biology, little is known about its gut microbiome. To investigate the gut bacterial community of A. grahami and explore its stability, diet and developmental stage were selected as the two variables. Larvae were reared on bovine liver, swine manure, and chicken manure, and high-throughput sequencing of the 1st, 2nd, and 3rd instar larvae and the newly emerged adults was performed. According to our results, the alpha diversity of the gut bacterial community did not significantly vary among different diets and developmental stages. Principal coordinate analysis revealed that the gut microbiome of A. grahami clustered together among different diets and developmental stages. The main phyla in the gut microbial community of A. grahami were Firmicutes and Proteobacteria, and the dominant genera were Vagococcus, Providencia, Lactobacillus, and Morganella. These findings characterized the gut microbiome of A. grahami and demonstrated that the gut bacterial community is fairly stable. The dominant genera Vagococcus, Providencia, Lactobacillus, and Morganella have the potential to serve as core microbiomes within the gut microbial community of A. grahami.

Chronic exposure to 2,2′-azobis-2-amidinopropane that induces intestinal damage and oxidative stress in larvae of Drosophila melanogaster

Embryonic development is exceptionally susceptible to pathogenic, chemistry and mechanical stressors as they can disrupt homeostasis, causing damage and impacted viability. Oxidative stress has the capacity to induce alterations and reshape the environment. However, the specific impacts of these oxidative stress-induced damages in the gastrointestinal tract of Drosophila melanogaster larvae have been minimally explored. This study used 2,2-azobis (2-amidinopropane) dihydrochloride (AAPH), a free radical generator, to investigate oxidative stress effects on Drosophila embryo development. The results showed that exposing Drosophila eggs to 30 mM AAPH during 1st instar larva, 2nd instar larva and 3rd instar larva stages significantly reduced hatching rates and pupal generation. It increased the activity of antioxidant enzymes and increased oxidative damage to proteins and MDA content, indicating severe oxidative stress. Morphological changes in 3rd individuals included decreased brush borders in enterocytes and reduced lipid vacuoles in trophocytes, essential fat bodies for insect metabolism. Immunostaining revealed elevated cleaved caspase 3, an apoptosis marker. This evidence validates the impact of oxidative stress toxicity and cell apoptosis following exposure, offering insights into comprehending the chemically induced effects of oxidative stress by AAPH on animal development.