Larval Tissues Research Articles

Potential Toxicity of <i>Indigofera tinctoria</i> Synthesized Iron Nanoparticles against <i>Aedes aegypti</i>

Mosquitoes are the most important single category of insects, killing millions of people worldwide each year by spreading a variety of diseases. The principal dengue vector Aedes aegypti, is expected to infect 2.5 billion people worldwide, or more than 40% of the world’s population. According to a WHO survey, 50-100 million cases are reported globally each year. Extensive fumigation of synthetic pesticides to control the mosquito vector in Pakistan during each post-monsoon season greatly increased environmental contamination and the loss of beneficial insects from urban environments. This study looked into the larvicidal and pupicidal efficiency of green synthesized iron nanoparticles against Ae. aegypti. Nanoparticles were subjected to several analyses, including UV-Vis, FTIR, FESEM, EDAX, XRD, Zeta Potential, and DLS. Ae. aegypti, the predominant dengue mosquito, was studied for its larvicidal and pupicidal activities. Indigofera tinctoria produced iron nanoparticles with LC50 values ranging from 4.468 ppm (I instar larvae) to 7.952 ppm (pupae). Laboratory experimental studies on larval body tissues, particularly fat cells, fingernail skin and midgut have been carried out. It has been determined the plant synthesis of iron nanoparticles are harmful to Ae. aegypti larval.

Ectopic expression in commonly used transgenic Drosophila GAL4 driver lines.

Transgenic tools such as the GAL4/UAS system in Drosophila have been used extensively to induce spatiotemporally controlled changes in gene expression and tissue-specific expression of a range of transgenes. We previously discovered unexpected expression of the commonly used dilp2-GAL4 line in tracheal tissue which significantly impacted growth phenotypes. We realized that few GAL4 lines have been thoroughly characterized, particularly when considering transient activity that may have significant impact on phenotypic readouts. Here, we characterized a further subset of 12 reportedly tissue-specific GAL4 lines commonly used in genetic studies of development, growth, endocrine regulation, and metabolism. Ten out of 12 GAL4 lines exhibited ectopic activity in other larval tissues, with seven being active in the larval trachea. Since this ectopic activity may result in phenotypes that do not depend on the manipulation in the intended target tissue, it is recommended to carefully analyze the outcome while taking this aspect into consideration.

Macrophages play a nutritive role in post-metamorphic maturation in Drosophila.

In the body of multicellular organisms, macrophages play an indispensable role in maintaining tissue homeostasis by removing old, apoptotic and damaged cells. In addition, macrophages allow significant remodeling of body plans during embryonic morphogenesis, regeneration and metamorphosis. Although the huge amount of organic matter that must be removed during these processes represents a potential source of nutrients, their further use by the organism has not yet been addressed. Here, we document that, during metamorphosis, Drosophila larval adipose tissue is infiltrated by macrophages, which remove dying adipocytes by efferocytosis and engulf leaking RNA-protein granules and lipids. Consequently, the infiltrating macrophages transiently adopt the adipocyte-like metabolic profile to convert remnants of dying adipocytes to lipoproteins and storage peptides that nutritionally support post-metamorphic development. This process is fundamental for the full maturation of ovaries and the achievement of early fecundity of individuals. Whether macrophages play an analogous role in other situations of apoptotic cell removal remains to be elucidated.

Toxocara canis infection in multiple types of animals: ophthalmological and pathological observations.

Human ocular toxocariasis (OT), caused by pet roundworm Toxocara canis (Nematoda Ascaridoidea), is a worldwide ocular parasitic infection that poses a severe threat to eyesight, especially in school-aged children. However, the infection process and pathological mechanism of Toxocara are difficult to study in the human body. This study was designed to explore long-term ocular manifestations in different rodents infected with Toxocara canis, uncovering the specific pathological mechanism and migration pathway of larvae after infection. The three types of experimental animals we selected were C57BL/6 mice, Mongolian gerbils and Brown Norway rats. Mice were randomly divided into five groups and infected orally with 1000, 2000, 4000, 8000 and 10,000 T. canis eggs; gerbils were randomly divided into four groups and infected orally with 1000, 2000, 4000 and 10,000 T. canis eggs; rats were randomly divided into three groups and infected orally with 2000, 6000 and 10,000 T. canis eggs. Their ocular changes were closely observed and recorded for at least 2months. We also enucleated the eyeballs of some animals to perform pathological sectioning and hematoxylin-eosin staining. After 3 dpi (days post-infection), hemorrhagic lesions, mechanical injury of the retina and larval migration could be observed in some infected animals. The ocular infection and mortality rates tended to be stable at 7 dpi. Larval tissue, structure disorder and inflammation could be observed in the pathological sections. In conclusion, the mice infected with 2000T. canis eggs and gerbils infected with 1000, 2000 and 4000T. canis eggs showing obvious ocular lesions and lower mortality rates could provide a basis for long-term observation.

The BTB transcription factor, Abrupt, acts cooperatively with Chronologically inappropriate morphogenesis (Chinmo) to repress metamorphosis and promotes leg regeneration

Many insects undergo the process of metamorphosis when larval precursor cells begin to differentiate to create the adult body. The larval precursor cells retain stem cell-like properties and contribute to the regenerative ability of larval appendages. Here we demonstrate that two Broad-complex/Tramtrack/Bric-à-brac Zinc-finger (BTB) domain transcription factors, Chronologically inappropriate morphogenesis (Chinmo) and Abrupt (Ab), act cooperatively to repress metamorphosis in the flour beetle, Tribolium castaneum. Knockdown of chinmo led to precocious development of pupal legs and antennae. We show that although topical application of juvenile hormone (JH) prevents the decrease in chinmo expression in the final instar, chinmo and JH act in distinct pathways. Another gene encoding the BTB domain transcription factor, Ab, was also necessary for the suppression of broad (br) expression in T. castaneum in a chinmo RNAi background, and simultaneous knockdown of ab and chinmo led to the precocious onset of metamorphosis. Furthermore, knockdown of ab led to the loss of regenerative potential of larval legs independently of br. In contrast, chinmo knockdown larvae exhibited pupal leg regeneration when a larval leg was ablated. Taken together, our results show that both ab and chinmo are necessary for the maintenance of the larval tissue identity and, apart from its role in repressing br, ab acts as a crucial regulator of larval leg regeneration. Our findings indicate that BTB domain proteins interact in a complex manner to regulate larval and pupal tissue homeostasis.

Induction of Oxidative Stress by Waterborne Copper and Arsenic in Larvae of European Seabass (Dicentrarchus labrax L.): A Comparison with Their Effects as Nanoparticles.

The aim of this work was to compare the potential induction of oxidative stress and the antioxidant enzymatic response after a short-term waterborne exposure to copper (Cu) and arsenic (As) with that of the nanoparticles (NPs) of these elements (Cu-NPs and As-NPs) in fish larvae of the species Dicentrarchus labrax. Larvae were grouped in several tanks and exposed to different concentrations of contaminants (0 to 10 mg/L) for 24 or 96 h under laboratory conditions. Copper and arsenic concentrations were analysed in larval tissues using ICP-MS. A set of oxidative stress biomarkers, including the levels of hydroperoxides (HPs), and superoxide dismutase (SOD) and catalase (CAT) activities were assessed. The trace element concentrations (mg/kg d.w.) in larvae ranged as follows: 3.28-6.67 (Cu at 24 h) and 2.76-3.42 (Cu at 96 h); 3.03-8.31 (Cu-NPs at 24 h) and 2.50-4.86 (Cu-NPs at 96 h); 1.92-3.45 (As at 24 h) and 2.22-4.71 (As at 96 h); and 2.19-8.56 (As-NPs at 24 h) and 1.75-9.90 (As-NPs at 96 h). In Cu tests, the oxidative damage (ROOH levels) was induced from 0.1 mg/L at both exposure times, while for Cu-NPs, this damage was not observed until 1 mg/L, which was paralleled by concomitant increases in SOD activity. The CAT activity was also increased but at lower metal concentrations (0.01 mg/L and 0.1 mg/L for both chemical forms). No oxidative damage was observed for As or As-NPs after 24 h, but it was observed for As after 96 h of treatment with 0.01 mg/L. A decrease in SOD activity was observed for As after 24 h, but it turned out to be increased after 96 h. However, As-NPs did not alter SOD activity. The CAT activity was stimulated only at 96 h by As and at 24 h by As-NPs. Therefore, the two chemical forms of Cu exhibited a higher bioaccumulation and toxicity potential as compared to those of As. Importantly, the association of both Cu and As in NPs reduced the respective trace metal bioaccumulation, resulting also in a reduction in the toxic effects (mortality and biochemical). Furthermore, the assessment of oxidative stress-related biomarkers in seabass larvae appears to be a useful tool for biomonitoring environmental-occurring trace elements.

Exploring Ocimum basilicum's Secondary Metabolites: Inhibition and Molecular Docking against Rhynchophorus ferrugineus for Optimal Action.

The objective of our work is to create a practical procedure to produce in vitro cell suspensions of O. basilicum and to ascertain the factors that encourage enhanced secondary metabolite production. We investigated the impact of these metabolites on Rhynchophorus ferrugineus's adult and larval target enzymes. The explants were cultivated on Murashige and Skoog (MS) media with 0.1 to 1 mg/L plant growth regulators (PGRs) to create calluses. 2,4-Dichlorophenoxyacetic acid (2,4-D), kinetin, 1-naphthylacetic acid (NAA), and indole-3-butryic acid (IBA) at 0.5, 0.5, 0.1, and 1 mg/L, respectively, with 3% sucrose led to the highest biomass accumulation. In cell suspensions, the total phenolic content (TPC) and total flavonoid content (TFC) were 39.68 and 5.49 mg/g DW, respectively, with abiotic Verticillium dahliae as an activator. Rosmarinic acid, ursolic acid, nepetoidin A and B, salvigenin, and quercetin-3-O-rutinoside as flavonoids and phenolics were analyzed using UPLC-I TQD MS, with the highest concentrations reached after 40 days. The extract demonstrates insecticidal activity against the fourth-instar larvae of R. ferrugineus, with adults at 1197 µg/mL and 12.5 µg/larvae as LC50 and LD50 values. The extract inhibited acetylcholine esterase (AChE), acid phosphatases (ACPs), alkaline phosphatases (ALPs), and gamma-aminobutyric acid-transaminase (GABA-T) in larval tissue in vitro, with IC50 values of 124.2, 149.3, 157.8, and 204.8 µg/mL, and in vivo, with IC50 values of 157.2, 179.4, 185.3, and 241.6 µg/mL, after 24 h. Pure compounds identified the activity of the extract, showing the inhibition of AChE, ACPs, ALPs, and GABA-T with IC50 values ˂ 200 µg/mL (in vitro). The ABMET examination revealed good oral permeability, and docking tests showed that the compounds bind AChE, ACPs, ALPs, and GABA-T. These findings show that a green bioprocessing method such as an O. basilicum cell suspension is a quick and straightforward technique for producing phenolic compounds, and it may be used to develop sustainable bio-insecticides and new green procedures.

Acclimation of intertidally reproducing sea-snails protects embryos from lethal effects of transient hyperthermia.

Embryos of Ilyanassa obsoleta (from Massachusetts and Florida) and Phrontis vibex (from Florida) were exposed to temperatures from 33to 37°C. In both species, very young embryos are especially sensitive to thermal stress. Brief early heat shock did not disturb spiral cleavage geometry but led to variable, typically severe defects in larval morphogenesis and tissue differentiation. In Ilyanassa but not P. vibex, early heat shock resulted in immediate slowing or arrest of interphase progression during early cleavage. This reversible arrest was correlated with improved prognosis for larval development and (in Massachusetts snails, at least) depended on parental acclimation to warm temperature (~25.5°C). Embryos from Massachusetts snails housed at lower temperature (16°C) exhibited cytokinesis failure when briefly incubated at 33°C during early cleavage, and tissue differentiation failure during incubation at 33°C begun at later stages. This preliminary study reveals a case in which stress-conditioned parents may endow embryos with protection against potentially lethal thermal stress during the most vulnerable stages of life.

Insecticidal activity and possible modes of action of secondary metabolites of some fungal strains and wild plants as natural pesticides against Spodoptera frugiperda

BackgroundThe fall armyworm (FAW), Spodoptera frugiperda, is an invasive and destructive pest to certain strategic crops, especially maize in Egypt. This research was conducted mainly to obtain secondary metabolites from some wild plants and fungal strains, use them in controlling FAW, and investigate their mode of action.ResultsThe ethyl acetate extracts of Cladosporium cladosporioides and Verticillium lecanii, as well as the alkaloid extracts of Ricinus communis and Nicotiana glauca, were extracted to obtain their secondary metabolites. The secondary metabolite contents were identified by gas chromatography mass spectrometry and NMR. The toxicity of all extracts against the 3rd-instar larvae of FAW was evaluated. The possible mode of action of the extracts was studied via their effects on larval enzyme activities and larval tissue. The toxicity results illustrated that, the extract of C. cladosporioides was more effective with LC50 229 ppm, followed by the extract of V. lecanii with LC50 341 ppm and N. glauca with LC50 404 ppm, while the least effective extract was R. communis with LC50 1110 ppm after 72 h of treatment. While, the results of larval enzyme activities showed that C. cladosporioides, V. lecanii, and R. communis extracts led to significant activity of AST, ALT, ACP, and ALP enzymes, but GST and AchE were inhibited in treated larvae compared with control. While N. glauca alkaloid extract caused significant inhibition of AST, ALT, ACP, AchE, and GST enzymes, ALP was activated in the treated larvae compared with control. The results of larval tissue slides indicated that the most affected tissues were the cuticle layer and the membrane lining of the midgut, in addition to the fatty bodies.ConclusionThus, natural pesticides would have a promising role in terms of controlling the FAW and according to this study, it was recommended that, alkaloid extracts of tested wild plants and ethyl acetate extracts of fungal strains be used as natural pesticides to control the fall armyworm, S. frugiperda.

Genetic diversity of black soldier flies in Vietnam based on DNA COI sequence

Abstract. Nguyen TN, Tran TBN, Tran HN. 2023. Genetic diversity of black soldier flies in Vietnam based on DNA COI sequence. Biodiversitas 24: 6727-6732. The Black Soldier Fly (BSF), Hermetia illucens (Linnaeus, 1758), is found in temperate and tropical regions of the world. In recent years, BSF has been cultivated as fodder in some areas of Vietnam. The genetic diversity of BSF in Vietnam has not yet been assessed. This study aimed to investigate the nucleotide diversity in COI sequences of BSF in Vietnam. We examined the COI nucleotide sequences of 22 BSFs using flies or larval tissue samples for DNA extraction and subsequent sequencing. Our analysis showed that the nucleotide composition of BSF consisted of 25.36% A, 36.04% T, 21.25% C, and 17.35% G, with a significant prevalence of A+T nucleotides of 61.40% compared to 38.60% for G+C nucleotides. Among 22 individual BSFs samples collected from ten provinces and cities, spanning seven agro-ecological regions in Vietnam, we identified 9 different haplotypes, which were distributed in five clades but mainly clustered in two main clades: E and F. Clade E, prominently represented by branch E, showed a higher abundance in Asia, while Clade F has been identified in Europe, Uganda, South Korea, Oceania and North America. Our results highlight the relatively high genetic diversity of the Vietnamese BSF population. In addition, the COI nucleotide sequences of these Vietnamese BSF samples were similar to those of Asian and Australian countries, showing significant similarities to the Palearctic and Oriental regions. These results provide valuable insights to inform BSF breeding and development in Vietnam.

Piwi expressed in Drosophila adipose tissues regulates systemic IGF signaling and growth via IGF-binding protein

Piwi and its partner, Piwi-interacting RNA (piRNA), are pivotal in suppressing the harmful effects of transposable elements (TEs) linked to genomic insertional mutagenesis. While primarily active in Drosophila's adult gonadal tissues, causing sterility in its absence, Piwi's role in post-embryonic development remains unclear. Our study reveals Piwi's functional presence in the larval fat body, where it governs developmental growth through systemic insulin/insulin-like growth factor (IGF) signaling (IIS). Piwi knockdown in the fat body resulted in dysregulated TE expression, reduced developmental rate and body growth, and diminished systemic IIS activity. Notably, Piwi knockdown increased Imaginal Morphogenic Protein Late 2 (Imp-L2) expression, akin to insulin-like growth factor-binding protein 7 (IGFBP7), reducing systemic IIS and inhibiting body growth. This unveils a novel role for Piwi in larval adipose tissues, emphasizing its importance in regulating systemic IIS and overall organismal growth.

Characterization of genetic and molecular tools for studying the endogenous expression of Lactate dehydrogenase in Drosophila melanogaster.

Drosophila melanogaster larval development relies on a specialized metabolic state that utilizes carbohydrates and other dietary nutrients to promote rapid growth. One unique feature of the larval metabolic program is that Lactate Dehydrogenase (Ldh) activity is highly elevated during this growth phase when compared to other stages of the fly life cycle, indicating that Ldh serves a key role in promoting juvenile development. Previous studies of larval Ldh activity have largely focused on the function of this enzyme at the whole animal level, however, Ldh expression varies significantly among larval tissues, raising the question of how this enzyme promotes tissue-specific growth programs. Here we characterize two transgene reporters and an antibody that can be used to study Ldh expression in vivo. We find that all three tools produce similar Ldh expression patterns. Moreover, these reagents demonstrate that the larval Ldh expression pattern is complex, suggesting the purpose of this enzyme varies across cell types. Overall, our studies validate a series of genetic and molecular reagents that can be used to study glycolytic metabolism in the fly.

Screening of insect immune suppressors using a recombinant phospholipase A2 of a lepidopteran insect.

Phospholipase A2 (PLA2 ) catalyzes phospholipids at the sn-2 position to release free fatty acids, including arachidonic acid (AA) or its precursor. The free AA is then oxygenated into different eicosanoids, which mediate the diverse physiological processes in insects. Any inhibition of the PLA2 catalysis would give rise to serious malfunctioning in insect growth and development. An onion moth, Acrolepiopsis sapporensis, encodes four different PLA2 genes (As-PLA2 A-As-PLA2 D), in which As-PLA2 A is dominantly expressed at all developmental stages and in different larval tissues. RNA interference of the As-PLA2 A expression significantly reduced the PLA2 activity of A. sapporensis, which suffered from immunosuppression. A recombinant As-PLA2 A protein was purified from a bacterial expression system, which exhibited a typical Michaelis-Menten kinetics and hence susceptible to a specific inhibitor to sPLA2 and dithiothreitol. A total of 19 bacterial metabolites derived from Xenorhabdus and Photorhabdus were screened against the recombinant As-PLA2 A. Five potent metabolites were highly inhibitory and followed a competitive enzyme inhibition. These five inhibitors suppressed the immune responses of A. sapporensis by inhibiting hemocyte-spreading behavior and phenoloxidase activity. However, an addition of AA could significantly rescue the immunosuppression induced by the selected inhibitors. These studies suggest that the recombinant As-PLA2 A protein can be applied for high-throughput screening of insect immunosuppressive compounds.

Analyzing the evolution of insect TMED gene and the expression pattern of silkworm TMED gene

Transmembrane emp24 domain (TMED) gene is closely related to immune response, signal transduction, growth and disease development in mammals. However, only the Drosophila TMED gene has been reported on insects. We identified the TMED family genes of silkworm, Tribolium castaneum, tobacco moth and Italian bee from their genomes, and found that the TMED family gene composition patterns of one α-class, one β-class, one δ-class and several γ-classes arose in the common ancestor of pre-divergent Hymenoptera insects, while the composition of Drosophila TMED family members has evolved in a unique pattern. Insect TMED family γ-class genes have evolved rapidly, diverging into three separate subclasses, TMED6-like, TMED5-like and TMED3-like. The TMED5-like gene was lost in Hymenoptera, duplicated in the ancestors of Lepidoptera and duplicated in Drosophila. Insect TMED protein not only has typical structural characteristics of TMED, but also has obvious signal peptide. There are seven TMED genes in silkworm, distributed in six chromosomes. One of seven is single exon and others are multi-exons. The complete open reading frame (ORF) sequences of seven TMED genes of silkworm were cloned from larval tissues and registered in GenBank database. BmTMED1, BmTMED2 and BmTMED6 were expressed in all stages and tissues of the silkworm, and all genes were expressed in the 4th and 5th instar and silk gland of the silkworm. The present study revealed the composition pattern of TMED family members, their γ class differentiation and their evolutionary history, providing a basis for further studies on TMED genes in silkworm and other insects.

Neochloris aquatica induces larval mortality, molting defects, and unstable flightless adults in the Asian tiger mosquito

The Asian tiger mosquito, Aedes albopictus, is a highly invasive and aggressive species capable of transmitting a large number of etiological agents of medical and veterinary importance, posing a high risk for the transmission of emerging viruses between animals and humans. In this work, we evaluated the mosquitocidal activity of Neochloris aquatica against A. albopictus throughout its development and analyzed whether this effect was potentiated when the microalga was cultivated under stress conditions due to nutrient deprivation. Our results suggest that N. aquatica produces metabolites that have negative effects on these insects, including larval mortality, interruption of pupal development, and incomplete emergence of adults when fed on microalgae in the larval stages. When microalgae were cultured under stress conditions, an increase in molting defects was recorded, and the number of healthy adults emerged drastically decreased. Histological studies revealed severe signs of total disintegration of different tissues and organs in the thorax and abdomen regions. The muscles and fat bodies in the midgut and foregut were severely distorted. In particular, larval intestinal tissue damage included vacuolization of the cytoplasm, destruction of brush border microvilli, and dilation of the intercellular space, which are distinctive morphological characteristics of apoptotic cells. Evidence suggests that N. aquatica produces metabolites with mosquitocidal effects that affect development and, therefore, the ability to vector etiological agents of medical and veterinary importance.

Developmental toxicity and neurobehavioral effects of sodium selenite and selenium nanoparticles on zebrafish embryos

Selenium, a trace mineral, is essential for several physiological processes in humans and animals. It is an antioxidant vital for the immunological response, DNA synthesis, thyroid hormone metabolism, and antioxidant defense enzymes. Zebrafish embryos and larvae were exposed to different concentrations of sodium selenite (SodSe) and selenium nanoparticles (SeNs) at various developmental stages. The study evaluated the impact of SodSe and SeNs on larvae survival, hatching rate, and morphological abnormalities. Also, acridine orange staining was used to analyze the apoptotic cell death, and behavioral tests were conducted to assess anxiety-like behaviors. The results showed that both SodSe and SeNs influence the development and neurobehavior of zebrafish larvae in a concentration-dependent manner. SodSe at high concentration causes low survival rates, delayed hatching, and increased morphological defects in zebrafish larvae. In addition, exposure to SodSe resulted in elevated apoptosis in different larval tissues. Zebrafish larvae treated with SodSe and SeNs exhibited anxiety-like behaviour, increased thigmotaxis, less exploratory behaviour, and less swimming patterns. The nerve conductions and stimuli responses evaluated through acetylcholine esterase (AChE) and cortisol assays, revealed a decrease in the activity in a dose-dependent manner of SodSe and SeNs. Interestingly, the effects of SeNs were lower even at higher concentrations when compared with SodSe at lower concentrations on zebrafish embryos. This shows that SeNs synthesized through biological methods may be less toxic and may have lower effect on the development and neurobehavior of zebrafish larvae. Thus, our study confirms the cytotoxic and neurobehavioral effects of SodSe and suggests the use of SeNs at lower concentration to provide insights into better understanding of developmental stages and metabolic pathways in zebrafish larvae.

Phenolics from Nigella sativa L. straw: Characterization and insecticidal activity against Agrotis ipsilon (Hüfnagel)

The excessive production of food and agro-waste has become a significant problem for society, the economy, and the environment. To meet the growing demand for food free from harmful synthetic insecticides, a recent study has investigated the potential use of an ethanolic extract obtained from the straw of Nigella sativa L., a byproduct of seed collection, as a bioinsecticide. The study also evaluated its in-vitro and in-silico acetylcholinesterase (AChE) inhibitory potential against the Agrotis ipsilon (Hufnagel) moth species, which is known to cause damage to various crops and ornamental plants. The high-performance liquid chromatography examination revealed that the ethanolic N. sativa straw extract contained 18 phenolics, including 3 simple phenols, 8 phenolic acids, and 7 flavonoids. Catechol (330.14 μg/ml), chlorogenic (169.23 μg/ml), and gallic (110.93 μg/ml) acids were the predominant phenolics. On the other hand, catechin (94.07 μg/ml), naringenin (91.99 μg/ml), and rutin (78.16 μg/ml) were the major flavonoids identified in the extract. The insecticidal activity of the extract against the 4th larval instar of A. ipsilon was evaluated using four concentrations (1.25–10 %). The study found that higher extract concentrations led to increased mortality in the larvae. Specifically, the concentration of 10 % resulted in the highest mortality rate of 96.67 %. Lower concentrations of 5 %, 2.5 %, and 1.25 % resulted in mortality rates of 51.11 %, 18.89 %, and 9.17 %, respectively. The extract also showed higher activity against AChE in larval tissue, with an inhibition percentage of 65.2 % after 24 h of treatment. Docking experiments confirmed that ellagic acid and apigenin had higher binding affinity than the control (lanate). These results demonstrate the potential of utilizing agricultural waste like N. sativa straw to create innovative and sustainable bioinsecticides.

Aneuploidy during development in facultative parthenogenetic Drosophila

From concatenated chromosomes to polyploidization, large-scale genome changes are known to occur in parthenogenetic animals. Here, we report mosaic aneuploidy in larval brains of facultatively parthenogenetic Drosophila. We identified a background of aneuploidy in D. mercatorum strains and found increased levels of aneuploidy in the larval brain tissue of animals arising parthenogenetically versus those arising from sexual reproduction. There is also intra-individual variation in germline-derived aneuploidy within the same strain. To determine if this is a general feature of facultative parthenogenesis in drosophilids, we compared sexually reproduced and parthenogenetic offspring from an engineered facultative parthenogenetic strain of D. melanogaster. In addition to germline-derived aneuploidy, this revealed somatic aneuploidy that increased by up to fourfold in parthenogens compared to sexually reproduced offspring. Therefore, the genetic combination identified in D. mercatorum that causes facultative parthenogenesis in D. melanogaster results in aneuploidy, which indicates that the loss of mitotic control resulting in parthenogenesis causes subsequent genome variation within the parthenogenetic offspring. Our findings challenge the assumption that parthenogenetic offspring are near genetic replicas of their mothers.

Differential expression of immunity-related genes in larval Manduca sexta tissues in response to gut and systemic infection.

The midgut epithelium functions as tissue for nutrient uptake as well as physical barrier against pathogens. Additionally, it responds to pathogen contact by production and release of various factors including antimicrobial peptides, similar to the systemic innate immune response. However, if such a response is restricted to a local stimulus or if it appears in response to a systemic infection, too is a rather underexplored topic in insect immunity. We addressed the role of the midgut and the role of systemic immune tissues in the defense against gut-borne and systemic infections, respectively. Manduca sexta larvae were challenged with DAP-type peptidoglycan bacteria - Bacillus thuringiensis for local gut infection and Escherichia coli for systemic stimulation. We compared the immune response to both infection models by measuring mRNA levels of four selected immunity-related genes in midgut, fat body, hematopoietic organs (HOs), and hemocytes, and determined hemolymph antimicrobial activity. Hemocytes and HOs were tested for presence and distribution of lysozyme mRNA and protein. The midgut and circulating hemocytes exhibited a significantly increased level of lysozyme mRNA in response to gut infection but did not significantly alter expression in response to a systemic infection. Conversely, fat body and HOs responded to both infection models by altered mRNA levels of at least one gene monitored. Most, but not all hemocytes and HO cells contain lysozyme mRNA and protein. These data suggest that the gut recruits immune-related tissues in response to gut infection whereas systemic infections do not induce a response in the midgut. The experimental approach implies a skewed cross-talk: An intestinal infection triggers immune activity in systemic immune organs, while a systemic infection does not elicit any or only a restricted immune response in the midgut. The HOs, which form and release hemocytes in larval M. sexta, i) synthesize lysozyme, and ii) respond to immune challenges by increased immune gene expression. These findings strongly suggest that they not only provide phagocytes for the cellular immune response but also synthesize humoral immune components.

Effects of acute and chronic chromium stress on the expression of heat shock protein genes and activities of antioxidant enzymes in larvae of Orthetrum albistylum

The dragonfly species Orthetrum albistylum, can accumulate heavy metals from its aquatic environment and thus serves as a biological indicator for monitoring and evaluating water quality. Heat shock proteins (HSPs) play important biological roles in resistance to various types of environmental stress. The full-length cDNA sequences of the heat shock cognate (hsc) 70 and heat shock protein (hsp) 70 genes were cloned from O. albistylum larvae. Relative levels of expression of hsc70 and hsp70 in the head, epidermis, midgut, and adipose tissue were measured by qRT-PCR after chronic and acute contamination of 5–8 instar larvae with chromium (Cr) solution, and under control conditions. Activities of superoxide dismutase (SOD) and catalase (CAT) in chronically contaminated larvae were also measured. Phylogenetic analysis revealed that the cloned hsc70 and hsp70 genes were highly homologous to known HSP70 family members reported in other insects. The mRNA levels of hsc70 and hsp70 did not differ significantly in various larval tissues. Under chronic chromium stress, hsc70 and hsp70 expression were upregulated to a maximum and then downregulated; hsp70 mRNA levels were higher than those of hsc70 at all concentrations of chromium. Under acute chromium stress, hsc70 expression was inhibited at low chromium concentrations and upregulated at chromium concentrations higher than 125 mg/L. However, hsp70 expression was higher than that in the control group and markedly higher than that of hsc70. Changes in SOD and CAT activities displayed consistent trends for different chronic chromium concentrations, first increasing and then decreasing over time. Collectively, these findings demonstrated the response of the HSP family of genes and antioxidant enzymes following exposure to heavy metal stress, as well as their potential applicability as biomarkers for monitoring environmental pollutants.