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Comparison of Comet assay profile for genotoxicity in larvae of Culex quiquefasciatus. (A) Control. (B) A. aspera. (C) Bti. (D) A. aspera + Bti.
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Five different weed plants viz. Convulvulus arvensis, Chenopodium murale, Tribulus terrestris, Trianthema portulacastrum, and Achyranthes aspera were investigated for their entomocidal and genotoxic effects against Culex quinquefasciatus mosquitoes. High mortality was observed at 72 hours in a dose dependent manner. Among all the tested plants, A....
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... assay. The comet assay was performed to detect the DNA damage in cells of treated larvae of Cx. quiquefasciatus with significant plant extract A. aspera and its combination with Bti. It was found that no DNA damage had occurred in treatment group compared to control (Fig. ...
Citations
... Among the potential secondary biological consequences of these pesticides, genotoxicity, and carcinogenicity are of special importance. Even the less toxic organophosphorus pesticides malathion and methyl parathion are reported to be genotoxic and carcinogenic 6,[22][23][24][25] . Our results indicated that, in the tested condition, malathion caused a significant micronucleus induction in mice at both 24 and 48 hours exposure periods. ...
Isolation of plant-based pesticides gaining recent interest due to the clastogenic implications of synthetic insecticides. The polyphenolic-rich fraction from the stem bark of Streblus asper was proven as an insecticide against Dysdercus cingulatus. This study aims to evaluate the genotoxicity of polyphenolic bioinsecticide from Streblus asper (PBSA) using the mouse bone marrow micronucleus assay and compare its effect with malathion, an organophosphorus insecticide, and vepacide, a neem-based bioinsecticide. The micronucleus assay was conducted after 24 and 48 hours after the second administration of the pesticides (two doses by i.p. injection for two consecutive days). Swiss albino mice were divided into six groups, each comprising six animals. The first group received dimethyl sulphoxide (Group I-DMSO control), the second group (Group II-positive control) received 100mg cyclophosphamide/ Kg body weight, Group III received 276mg (LD 10
... Ae. aegypti mosquitoes were reared in insect cage using the protocol optimized by Zulhussnain et al. (2020) with 12 h day/night cycle under standard conditions (60 ± 10% RH and 26 ± 1°C) in Entomology Lab, Department of Zoology, Government College University Faisalabad. Grounded fish food and Purina cat food were used to feed newly hatched larvae and 5-8 days old larvae, respectively. ...
... DNA extraction was performed as described by Zulhussnain et al. (2020). Ae. aegypti samples were homogenized in 300 μl lysis buffer (2 mM EDTA, 0.4 M NaCl, and 10 mM Tris-HCL pH 8.0), 20% SDS (sodium dodecyl sulfate) and 100 μl Proteinase K (100 mg μl). ...
The vector role of Aedes aegypti for viral diseases including dengue and dengue hemorrhagic fever makes it imperative for its proper control. Despite various adopted control strategies, genetic control measures have been recently focused against this vector. CRISPR Cas9 system is a recent and most efficient gene editing tool to target the sex determination pathway genes in Ae. aegypti . In the present study, CRISPR Cas9 system was used to knockout Ae. aegypti doublesex ( Aaedsx ) and Ae. aegypti sexlethal ( AaeSxl ) genes in Ae. aegypti embryos. The injection mixes with Cas9 protein (333 ng ul ⁻¹ ) and gRNAs (each at 100 ng ul ⁻¹ ) were injected into eggs. Injected eggs were allowed to hatch at 26 ± 1°C, 60 ± 10% RH. The survival and mortality rate was recorded in knockout Aaedsx and AaeSxl . The results revealed that knockout produced low survival and high mortality. A significant percentage of eggs (38.33%) did not hatch as compared to control groups ( P value 0.00). Highest larval mortality (11.66%) was found in the knockout of Aaedsx female isoform, whereas, the emergence of only male adults also showed that the knockout of Aaedsx (female isoform) does not produce male lethality. The survival (3.33%) of knockout for AaeSxl eggs to the normal adults suggested further study to investigate AaeSxl as an efficient upstream of Aaedsx to target for sex transformation in Ae. aegypti mosquitoes.
... Genomic DNA was extracted from earthworms by the TNE (Tris-HCL-NaCl-EDTA) buffer method (Zulhussnain et al., 2020). Earthworms (prostomium, clitella, gut, and tail) were separately homogenized in 400 µl of TNE buffer (0.45 M NaCl, 3 mM EDTA, and 10 mM Tris-HCl; pH: 8.0). ...
... DNA variations of earthworms were determined by employing the RAPD-PCR assay (Zulhussnain et al., 2020). Six oligomer primers of Gene Link-A series were used for the amplification of DNA (Supplementary Table S1). ...
... The other three levels are intermediate between 0 and 4. Computer image analysis (Casp R software) was used to measure head length, tail length, comet length, head DNA, tail DNA, and tail movement. Cells with maximum damage showed the highest value for tail DNA and minimum value of head DNA (Attullah et al., 2020;Zulhussnain et al., 2020). ...
Sewage sludge, rich in organic matter and nutrients, is widely used as a fertilizer to increase the fertility of soil. The direct application of sewage sludge without any treatment causes soil contamination as well as significantly affects the earthworm population. In the present study, the effect of sewage sludge-amended soil on growth, enzyme activities, and the DNA damage in Pheretima posthuma earthworms was studied under wheat straw and biochar treatment of 30-day laboratory incubation. Wheat straw, biochar, and sewage sludge were applied at 0 (control), 5, 10, and 25% w/w along with combined treatment of sewage sludge with biochar and wheat straw, respectively at 25% each. After the incubation period, the percentage change in mean weight and length was measured and esterase and phosphatase enzyme activities were quantitatively determined. RAPD-PCR and the comet assay were performed to assess the genotoxicity. A significant weight loss was observed (26%) at a 25% rate of sewage sludge and biochar (11%). Similarly, a maximum decrease in length was observed with sewage sludge (2.5%) followed by biochar (0.80%) at a 25% application rate. Mean weight and length both decreased by increasing the rate of sewage sludge and biochar. In addition, no change was observed in the weight and length of P. posthuma in the treatment consisting of sewage sludge and wheat straw. Moreover, sewage sludge caused inhibition of esterase and phosphatase activities as well as induced DNA damage. The comet parameters showed that wheat straw and biochar ameliorated the toxic effects of sewage sludge. It is, therefore, concluded that sewage sludge has a tangible impact on earthworms which ultimately disrupts ecosystem functions and wheat straw and biochar can thus be utilized to reduce the toxicity of sewage sludge in Pheretima posthuma earthworms.
... Bakar et al. Phytochemical compounds in Achyranthes aspera extracts, such as saponins, tannins, steroids, glycosides, alkaloids, anthraquinones, and terpenoids, cause metabolic and neurotoxic damage in Cx. quinquefasciatus larvae (Zulhussnain et al. 2020). The authors suggested that these phytochemicals negatively modulate the enzymatic activities of insect esterase and phosphatase, such as AChE, AcP (acid phosphatase), AkP (alkaline phosphatase), α-carboxylesterases, and β-carboxylesterases. ...
Mosquitoes (Diptera: Culicidae) are insect vectors of epidemiologically important arboviruses owing to their behavior, physiology, morphology, and proximity to humans, which require incisive strategies to contain their spread. The failure of current arbovirus management plans and lack of fully effective treatments suggest that vector control by botanical insecticides could be an effective and safe strategy. Botanical insecticides are obtained from renewable sources and have complex chemical compositions, different modes of action, and selective toxicity for target organisms. In this review, we present the main control strategies for insects belonging to the genera Aedes, Culex, and Anopheles and discuss the possibility of using botanical insecticides in the integrated management of vectors. Numerous botanical insecticide formulations are presented, and their potential modes of action during the immature stages include damage to the egg exocorionic network and abnormal disruption of embryos, which result from deficiencies in egg chitinization, impairment of larval morphology, and inhibition or differential expression of enzymes, promoting changes in the digestive tract epithelium and reduced larval mobility, and impairment of external surfaces or the respiratory system of pupae, altering pupal swimming patterns. In adult insects, botanical insecticides can promote incomplete ecdysis, in addition to dysfunction of olfactory receptors, food traffic, and reproductive function. Thus, broad-spectrum botanical insecticides can be used to control the different stages of insect development. The contributions of nanotechnology to vector control should be further explored to enhance the insecticidal activity and stability of botanical insecticides under different conditions.
... This has prompted the integration of effective, environmentallyfriendly and nontoxic plant-based larvicides into mosquito control programmes (Allgeier et al., 2018). Several studies have highlighted the toxicity potentials of different plants to mosquitoes (Sillo et al., 2019;Zulhussnain et al., 2020;Attaullah et al., 2020). In a study, Karthi et al. (2020) described the larvicidal activity of crude and acetone extracts of Rhizophora mucronata against Cx. ...
... Also, obtained in this study was increased larvae mortality with increased concentrations of H. suaveolens extracts. Thus, this is in agreement with previous studies (Conti et al., 2012;Sultana et al., 2016;Raiz et al., 2018;Zulhussnain et al., 2020). ...
... These are bioactive compounds that play crucial role individually or H. K. Aremu et al. synergistically to cause mortality or delay larvae growth (Elumalai et al. 2013;Yadav et al., 2014;Pavela et al., 2018). It has been previously reported by Elumalai et al. (2017b) and Zulhussnain et al. (2020) that the presence of phytochemicals in leaves extracts of Leucas aspera and Convulvulus arvensis significantly contributed to their larvicidal and insecticidal potentials. ...
Vector control strategies have focused on the development of effective and ecofriendly alternatives. In the present study, investigation of larvicidal and genotoxic effects of leaves of Hyptis suaveolens from four different extraction solvents (aqueous, hexane, methanol and acetone) on fourth instar larvae of Culex quinquefasciatus was carried out. Extraction was done using soxhlet apparatus and the characteristics functional group of active constituents were identified using Fourier Transform Infrared spectrophotometer. Larvicidal activities were screened using three different concentrations (50, 150 and 250 mg/mL) following WHO standard protocol and mortality was recorded after 24, 48 and 72 hr. Hexane extract showed the highest mortality (27.92, 38.75, 90.42 %; LC50: 272.5, 191.3, 114.8 mg/mL), followed by aqueous extract (20.83, 34.58, 59.58 %; LC50: 496.6, 392.9, 208.1 mg/mL) and acetone extract (20.83, 32.08, 59.58 %; LC50: 1111.2, 393.6, 266.1 mg/mL) and methanol extract (17.92, 29.17, 52.92 %; LC50: 466.0, 400.1, 272.3 mg/mL). Enzyme profile such as superoxide dismutase (SOD), glutathione-S-transferase (GST), catalase (CAT), glutathione (GSH) and alkaline phosphatase (ALP) were significantly altered in the larvae exposed to the four extracts. Phytochemical screening of all solvents extract revealed the presence of saponins, flavonoids, terpenoids and alkaloids as common constituents. Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) profile implied hexane and aqueous extracts altered the DNA of larvae. Furthermore, FTIR Spectroscopic analysis revealed phenols, alcohols, aliphatic primary amines and saponins as the major groups in the extracts. Conclusively, this study established the lethal potential of extracts of H. suaveolens as alternative plant-based and eco-friendly larvicide against Cx. quinquefasciatus.
... This has prompted the integration of effective, environmentallyfriendly and nontoxic plant-based larvicides into mosquito control programmes (Allgeier et al., 2018). Several studies have highlighted the toxicity potentials of different plants to mosquitoes (Sillo et al., 2019;Zulhussnain et al., 2020;Attaullah et al., 2020). In a study, Karthi et al. (2020) described the larvicidal activity of crude and acetone extracts of Rhizophora mucronata against Cx. ...
... Also, obtained in this study was increased larvae mortality with increased concentrations of H. suaveolens extracts. Thus, this is in agreement with previous studies (Conti et al., 2012;Sultana et al., 2016;Raiz et al., 2018;Zulhussnain et al., 2020). ...
... These are bioactive compounds that play crucial role individually or H. K. Aremu et al. synergistically to cause mortality or delay larvae growth (Elumalai et al. 2013;Yadav et al., 2014;Pavela et al., 2018). It has been previously reported by Elumalai et al. (2017b) and Zulhussnain et al. (2020) that the presence of phytochemicals in leaves extracts of Leucas aspera and Convulvulus arvensis significantly contributed to their larvicidal and insecticidal potentials. ...
Vector control strategies have focused on the development of effective and ecofriendly alternatives. In the present study, investigation of larvicidal and genotoxic effects of leaves of Hyptis suaveolens from four different extraction solvents (aqueous, hexane, methanol and acetone) on fourth instar larvae of Culex quinquefasciatus was carried out. Extraction was done using soxhlet apparatus and the characteristics functional group of active constituents were identified using Fourier Transform Infrared spectrophotometer. Larvicidal activities were screened using three different concentrations (50, 150 and 250 mg/mL) following WHO standard protocol and mortality was recorded after 24, 48 and 72 hr. Hexane extract showed the highest mortality (27.92, 38.75, 90.42 %; LC50: 272.5, 191.3, 114.8 mg/mL), followed by aqueous extract (20.83, 34.58, 59.58 %; LC50: 496.6, 392.9, 208.1 mg/mL) and acetone extract (20.83, 32.08, 59.58 %; LC50: 1111.2, 393.6, 266.1 mg/mL) and methanol extract (17.92, 29.17, 52.92 %; LC50: 466.0, 400.1, 272.3 mg/mL). Enzyme profile such as superoxide dismutase (SOD), glutathione-S-transferase (GST), catalase (CAT), glutathione (GSH) and alkaline phosphatase (ALP) were significantly altered in the larvae exposed to the four extracts. Phytochemical screening of all solvents extract revealed the presence of saponins, flavonoids, terpenoids and alkaloids as common constituents. Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) profile implied hexane and aqueous extracts altered the DNA of larvae. Furthermore, FTIR Spectroscopic analysis revealed phenols, alcohols, aliphatic primary amines and saponins as the major groups in the extracts. Conclusively, this study established the lethal potential of extracts of H. suaveolens as alternative plant-based and eco-friendly larvicide against Cx. quinquefasciatus.
Plants store chemical defenses that act as toxins against herbivores, such as toxic isothiocyanates (ITCs) in Brassica plants, hydrolyzed from glucosinolate (GLS) precursors. The fitness of herbivorous larvae can be strongly affected by these toxins, causing immature death. We modeled this phenomenon using a set of ordinary differential equations and established a direct relationship between feeding, toxin exposure, and the net energy of a larva, where the fitness of an organism is proportional to its net energy according to optimal foraging theory. Optimal foraging theory is widely used in ecology to model the feeding and searching behavior of organisms. Although feeding provides energy gain, plant toxins and foraging cause energy loss for the larvae. Our equations explain that toxin exposure and foraging can sharply reduce larval net energy to zero at an instar. Since herbivory needs energy, the only choice left for a larva is to stop feeding at that time point. If that is significantly earlier than the end of the last instar stage, the larva dies without food. Thus, we show that plant toxins can cause immature death in larvae from the perspective of optimal foraging theory.
Citation: Petchidurai, G.; Sahayaraj, K.; Al-Shuraym, L.A.; Albogami, B.Z.; Sayed, S.M. Insecticidal Activity of Tannins from Selected Brown Macroalgae against the Cotton Leafhopper Amrasca devastans. Plants 2023, 12, 3188. https://doi. Abstract: Seaweeds, also known as marine macroalgae, are renewable biological resources that are found worldwide and possess a wide variety of secondary metabolites, including tannins. Drifted brown seaweed (DBSW) is particularly rich in tannins and is regarded as biological trash. The cotton leaf hopper Amrasca devastans (Distant) has caused both quantitative and qualitative losses in cotton production. Drifted brown seaweeds (DBSWs) were used in this study to extract, qualitatively profile, and quantify the levels of total tannins, condensed tannins, hydrolyzable tannins, and phlorotannins in the seaweeds; test their insecticidal activity; and determine the mechanism of action. The largest amount of tannin extract was found in Sargassum wightii Greville (20.62%) using the Soxhlet method (SM). Significantly higher amounts of hydrolyzable tannins (p = 0.005), soluble phlorotannins (p = 0.005), total tannins in the SM (p = 0.003), and total tannins in the cold percolation method (p = 0.005) were recorded in S. wightii. However, high levels of condensed tannins (CTAs) were observed in Turbinaria ornata (Turner) J. Agardh (p = 0.004). A. devastans nymphs and adults were examined for oral toxicity (OT) and contact toxicity (CT) against DBSW tannin crude extract and column chromatographic fractions 1 (Rf = 0.86) and 2 (Rf = 0.88). Stoechospermum polypodioides (J.V. Lamouroux) J. Agardh crude tannin was highly effective against A. devastans using the OT method (LC 50 , 0.044%) when compared with the standard gallic acid (LC 50 , 0.044%) and tannic acid (LC50, 0.122%). Similarly, S. wightii fraction 2 (LC 50 , 0.007%) showed a greater insecticidal effect against A. devastans adults in OT than gallic acid (LC 50 , 0.034%) and tannic acid (LC 50 , 0.022%). The mechanism of action results show that A. devastans adults treated with crude tannin of T. ornata had significantly decreased amylase, protease (p = 0.005), and invertase (p = 0.003) levels when compared with the detoxification enzymes. The levels of glycosidase, lactate dehydrogenase, esterase, lipase, invertase, and acid phosphate activities (p = 0.005) of S. wightii were reduced when compared with those of the Vijayneem and chemical pesticide Monocrotophos. In adult insects treated with LC 50 concentrations of S. wightii tannin fraction 1, the total body protein (9.00 µg/µL) was significantly reduced (OT, LC 50-0.019%). The SDS-PAGE analysis results also show that S. wightii tannin fraction 1 (OT and CT), fraction 2 (OT), and S. polypodioides fraction 2 (CT) had a significant effect on the total body portion level, appearance, and disappearance of some proteins and polypeptides. This study shows that the selected brown macroalgae can be utilized for the safer management of cotton leaf hoppers.
Plants store chemical defenses that act as toxins to herbivores, such as toxic isothiocyanates (ITCs) in Brassica plants, hydrolyzed from the precursor glucosinolates (GLSs). Fitness of herbivorous larvae can be highly affected by these toxins, causing immature deaths. Theoretically, toxins can inordinately reduce larval fitness to death. We model this phenomenon by a set of ordinary differential equations and establish a direct relationship between feeding, toxin exposure and net energy of a larva, where the fitness of an organism is proportional to its net energy according to the optimal foraging theory. Our equations explain that toxin exposure can steeply reduce larval net energy to zero at an instar stage. Since herbivory needs energy, the only choice left for a larva is to stop feeding at the time point. If that time-point is significantly earlier than the end of the last instar stage, the larva dies.
