Bioefficacy of ursolic acid and its derivatives isolated from Catharanthus roseus (L) G. Don leaf against Aedes aegypti, Culex quinquefasciatus, and Anopheles stephensi larvae.

The aim of the present study was to identify the active principle from Catharanthus roseus leaf using larvicidal bioassay against three mosquito species viz. Aedes aegypti, Culex quinquefasciatus, and Anopheles stephensi. Preliminary studies of the three successive extracts such as hexane, chloroform, and methanol against Ae. aegypti larvae showed that the chloroform extract was more active with LC50 and LC90 values of 40.09 ppm and 189.15 ppm respectively. Bioassay guided fractionation of the active chloroform extract resulted in the isolation of a triterpenoid (ursolic acid) as the active constituent. Three derivatives acetate, formate, and benzoate were prepared using this, and they were tested for their larvicidal activity against three mosquito species. The acetyl derivative was highly active against all the three species compared to the parent compound ursolic acid; the activities of benzoate and formate were higher than ursolic acid when tested against Cx. quinquefasciatus. This is the first report related to ursolic acid from C. roseus with mosquito larvicidal activity. The pure compound could be considered for medicinal and other pharmacological applications in future.

Valorization of chick feather wastes by Geobacillus thermodenitrificans PS41 to enhance the growth of Vigna unguiculata plant and Cyprinus carpio fish.

Chicken feather meal has had a significant biofertilizer approach in recent years. The current study aims to assess feather biodegradation to promote plant and fish growth. The Geobacillus thermodenitrificans PS41 strain was more efficient in feather degradation. Feather residues were separated after degradation and evaluated under a scanning electron microscope (SEM) to detect bacterial colonization on feather degradation. It was observed that the rachi and barbules were entirely degraded. The complete degradation by PS41 suggests a relatively more efficient feather degradation strain. According to Fourier-transform infrared spectroscopy (FT-IR) studies, PS41 biodegraded feathers contain the functional groups of aromatic, amine, and nitro compounds. The present study suggested that biologically degraded feather meal improved plant growth. The feather meal combined with nitrogen-fixing bacterial strain showed the highest efficiency. The biologically degraded feather meal and Rhizobium combination induced physical and chemical changes in the soil. It is directly involved in soil amelioration, plant growth substance, and soil fertility, enhancing a healthy crop environment. The feather meal 4 and 5% was used as a feed diet of common carp (Cyprinus carpio) to increase growth performances and feed utilization parameters. In hematological and histological studies of formulated diets, significantly no toxic effects occurred in fish blood, gut, or fimbriae.

Bioefficacy of isolated compound l-isoleucine, N-allyloxycarbonyl-, and dodecyl ester from entomopathogenic actinobacteria Actinokineospora fastidiosa against agricultural insect pests, human vector mosquitoes, and antioxidant activities.

Spodoptera litura and Helicoverpa armigera are polyphagous pests of agricultural crops in the Asian tropics since these pests have been responsible for massive crop and carry economic losses and low commodity production. At the same time, mosquitoes are vectors for numerous dreadful diseases, which is the most important group of insect for their public health concern. Using synthetic insecticides to control the pests can lead to contamination of land surface and groundwater and impact beneficial soil organisms and nontarget species. Applications of bioactive compounds are received considerable attention across the world as alternatives to synthetic insecticides. In the current study, actinobacterial secondary metabolite was isolated from Actinokineospora fastidiosa for the first time. The effect of actinobacterial metabolite (l-isoleucine, N-allyloxycarbonyl-, and dodecyl ester) was assessed on agricultural pest S. litura and H. armigera, mosquito vectors larvae Ae. aegypti, An. stephensi, and Cx. quinquefasciatus. The bioactive fraction was characterized through UV, FTIR, and NMR analysis. GC-MS analyses reveal the existence of a bioactive compound with a respective retention time of 19.740 responsible for larvicidal activity. The bioefficacy of the l-isoleucine, N-allyloxycarbonyl-, and dodecyl ester showed high antifeedant activity on S. litura (80.80%) and H. armigera (84.49%); and larvicidal activity on S. litura (82.77%) and H. armigera (88.00%) at 25 µg/mL concentration, respectively. The effective LC50 values were 8.07 µg/mL (F = 2.487, r2 = 0.988, P ≤ 0.05) on S. litura and 7.53 µg/mL (F = 123.25, r2 = 0.951, P ≤ 0.05) on H. armigera. The mosquito larvicidal effect of isolated compounds l-isoleucine, N-allyloxycarbonyl-, and dodecyl ester treated against Ae. aegypti, An. stephensi, and Cx. quinquefasciatus the obtained percentage mortality was 96.66, 83.24, 64.52, 50.00, and 40.00% against Ae. aegypti; 100.00, 86.22, 73.81, 65.37, and 56.24% against An. stephensi; 100.00, 90.00, 76.24, 68.75, and 56.23% against Cx. quinquefasciatus. The mosquito larvae of Ae. aegypti obtained LC50 value was 13.25 µg/mL, F = 28.50, r2 = 0.90; on An. stephensi was 10.19 µg/mL, F = 15.55, r2 = 0.83, and Cx. quinquefasciatus was 9.68 µg/mL, F = 20.00, r2 = 0.87. Furthermore, l-isoleucine-, N-allyloxycarbonyl-, and dodecyl ester-treated larvae produced significant pupicidal activity on S. litura (62.71%) and H. armigera (66.50%) at 25 µg/mL, along with increased larval and pupal duration as compared to control group. Treated larvae revealed obliteration in the midgut epithelial cells and destruction of microvilli was noticed as compared to the control. The isolated compounds l-isoleucine, N-allyloxycarbonyl-, and dodecyl ester did not produce any significant mortality on zebrafish embryos in all tested concentrations on biosafety observation. The potential microbial isolated molecule may fit well in IPM programs. Since the risk to human health, the environment, etc. is unknown.

Hematological changes, redox imbalance, and changes in Na+/K+-ATPase activity caused by bisphenol-A and the integrated biomarker responses in Labeo rohita (Hamilton, 1822).

Bisphenol-A (BPA) is a plasticizer commonly used in the plastics industry to manufacture plastic materials. It is abundant in aquatic ecosystems, resulting in increased contamination and lower concentrations that may represent a significant threat to the aquatic system. Hence in the present study, an Indian major carp, Labeo rohita, was exposed to two different BPA concentrations (1 and 10 µg/L) for 30 days. Compared to control, the chronic effects resulted in significant alterations in red blood cell (RBC) and white blood cells (WBC) count. The exposure to BPA caused significant changes in antioxidant activity in gill, liver, and kidney tissues (inferred by catalase, glutathione peroxidase, and glutathione S-transferase activity) in L. rohita. Regarding lipid peroxidation (LPO), we observed an increase in liver and kidney alteration, while LPO was noted in gill tissue compared to the control. Furthermore, increased Na+/K+-ATPase activity was observed in gills at the end of the 10th day and a gradual decrease at the end of the 30th day. These results indicated that exposure to BPA alters the RBC and WBC levels, antioxidant enzyme activity (gills, liver, and kidney), and Na+/K+-ATPase activity in the gill of L. rohita exposed to BPA (at 1 and 10 µg/L). Therefore, our findings will help us gain better insight into the toxicity of BPA in freshwater ichthyofauna.

Mosquito Larvicidal Activity of Compounds from Unripe Fruit Peel of Avocado (Persea americana Mill.).

Mosquitoes are important vectors responsible for spreading a number of diseases affecting both humans and animals. Many diseases as dengue, chikungunya, yellow fever, malaria, filariasis and Japanese encephalitis are spread by mosquitoes. There are many reports of plant extracts and their active constituents showing anti-mosquito activities as larvicidal, pupicidal, ovicidal and adulticidal activities. Persea americana Mill. (Lauraceae), known as avocado, has been reported to show many pharmacological and antimicrobial activities. In this communication, the mosquito larvicidal activities of the three-active constituents, avocadene, avocadyne and avocadenol-A, from the methanolic extract of the unripe fruit peel are presented. The three mosquito species studied were Aedes aegypti, Culex quinquefasciatus and Anopheles stephensi. All three compounds showed the highest larvicidal activity against An. stephensi, LC50 values being 2.80ppm for avocadene, 2.33ppm for avocadyne and 2.07ppm for avocadenol-A. Avocadene showed larvicidal activity of 3.73ppm against Ae. aegypti and 5.96ppm against Cx. quinquefasciatus. The LC50 value of avocadyne was 5.35ppm against Ae. aegypti and 3.98ppm against Cx. quinquefasciatus. Similarly, avocadenol-A showed 6.56ppm against Ae. aegypti and 2.35ppm against Cx. quinquefasciatus. The active constituents were isolated by bioactivity-guided fractionation by silica gel column chromatography and RP HPLC. The compounds were identified by physical and spectroscopic data and compared with literature values already reported.

Larvicidal, antioxidant and biotoxicity assessment of (2-(((2-ethyl-2 methylhexyl)oxy)carbonyl)benzoic acid isolated from Bacillus pumilus against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus.

Mosquitoes are a vector for many dreadful diseases known for their public health concern. The continued use of synthetic insecticides against vector control has led to serious environmental impacts, human health problems, and the development of insect resistance. Hence, alternative mosquito control methods are needed to protect the environment and human health. In the present study, the bioefficacy of (2-(((2-ethyl-2 methylhexyl)oxy)carbonyl) benzoic acid isolated from Bacillus pumilus were tested against Aedes aegypti, Culex quinquefasciatus and Anopheles stephensi. The isolated bioactive compound was characterized through thin layer chromatography (TLC), UV-visible spectroscopy (UV), Fourier-transform infrared spectroscopy, nuclear magnetic resonance spectroscopy, and gas chromatography-mass spectrometry analysis. The pure compound caused a high percent mortality rate in a dose-dependent manner, the obtained values were 96, 82, 69, 50 and 34%; 86, 72, 56, 43, and 44%; 100, 90, 83, 70 and 56% against Ae. aegypti, Cx. quinquefasciatus, and An. stephensi respectively. The effective lethal concentration values (LC50) were 13.65, 14.90 and 9.64 ppm against Ae. aegypti, Cx. quinquefasciatus, An. Stephensi, respectively. The effect of (2-(((2-ethyl-2 methylhexyl)oxy)carbonyl) benzoic acid significantly increased the superoxide dismutase, catalase, α, ß esterase and Glutathione-S-transferase level after 24 h of the treatment period. The comet assay confirmed that isolated compound causes DNA damage in all tested insects. Histopathological examinations of treated larvae showed shrunken body posture, damaged epithelial cells and microvillus as compared to control organisms. The biosafety of the isolated compound was assessed against G. affinis and did not produce mortality which confirmed that the activity of the isolated compound is species specific. The current study concludes that the critical success factors of new insecticidal agent development are based on the eco-compatibility and alternative tools for the pesticide producing industry.

Evaluation of antioxidant response and Na+-K+-ATPase activity in zebrafish exposed to polyethylene microplastics: Shedding light on a physiological adaptation.

Although the toxicity of microplastics has already been demonstrated in different animal models, our knowledge about the response of freshwater fish to this pollutant is still limited. Thus, we aimed to evaluate the impact of exposure of zebrafish adults (Danio rerio) to environmentally relevant concentrations of polyethylene microplastics (PE-MPs) (5 and 50 µg/L) and at different times of exposure (10 and 20 days). Initially, scanning electron microscope image illustrated size and format of the particle and FTIR analysis confirmed the presence of PE-MPs in the gastrointestinal tract of fish (at both concentrations tested). Subsequently, an alteration of oxidative and antioxidant responses was evaluated in the liver and brain. The results showed that catalase (CAT) activity, in liver, was significantly decreased, as was glutathione S-transferases (GSTs) activity (on the 10th experimental day). However, after 20 days of exposure, we observed a concentration-dependent increase in GST activity in liver of the animals exposed to PE-MPs. Furthermore, the lipid peroxidation (LPO) level was significantly increased by exposure to MPs, especially in the brain, after 20 days of exposure. The increase in Na+-K+-ATPase activity in the animals' gills was correlated with the increased production of malondialdehyde (MDA), which suggests the existence of a compensatory mechanism in which the high activity of this enzyme would be necessary to regulate the loss of ions caused by the increase in the processes of LPO, which has never been previously demonstrated. Thus, our study sheds light on a new physiological adaptation to deal with the oxidative effects of PE-MPs, in addition to supporting the future use of the assessment of Na+/K+-ATPase activity as a biomarker of the toxicity of these pollutants.

Green synthesis of silver nanoparticle using Leonotis nepetifolia and their toxicity against vector mosquitoes of Aedes aegypti and Culex quinquefasciatus and agricultural pests of Spodoptera litura and Helicoverpa armigera.

Pest insects causing damage to cultivable crops and food products by feeding, fecundity, and parasitizing livestock, also being a nuisance to human health. In consideration with human health, the World Health Organization reports that more than 50% of the world's population is presently at risk from mosquito-borne diseases. Mosquitoes are primary vectors for major dreadful diseases such as yellow fever, malaria, and dengue fever, which infect millions of human beings all over the world and kill millions of peoples every year. The present research work was carried out to evaluate the antifeedant, larvicidal, pupicidal, larval, and pupal duration activity of Leonotis nepetifolia-mediated silver nanoparticles (AgNPs) against Spodoptera litura, Helicoverpa armigera, Aedes aegypti, and Culex quinquefasciatus. Biosynthesized AgNPs were characterized through various techniques such as UV-Vis spectrometer, X-ray powder diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), transmission electron microscopy (TEM), dynamic light scattering (DLS), and zeta potential analysis. The AgNPs showed potential antifeedant activity of 78.77% and 82.16% against the larvae of S. litura and H. armigera, respectively. The maximum larval mortality rate (78.49% and 72.70%) and maximum pupal mortality rate (84.66% and 77.44%) were observed against S. litura and H. armigera. Mosquito larvae were tested with biosynthesized AgNPs, and recorded LC50 values were 47.44 ppm and 35.48 ppm on A. aegypti and C. quinquefasciatus, respectively. The histological examinations showed that the acceleration of the nanomaterial caused severe tissue damage in the epithelial and goblet cells in the larval midgut region of S. litura, H. armigera, A. aegypti, and C. quinquefasciatus. Biosynthesis of silver nanoparticles using L. nepetifolia is an ideal eco-friendly approach for the management of insect pests. Graphical abstract.

Larvicidal Activity of Elytraria acaulis against Culex quinquefasciatus and Aedes aegypti (Diptera: Culicidae).

BACKGROUND: Mosquitoes are blood sucking arthropods and serve as vectors of many diseases causing serious health problems to human beings. Culex quinquefasciatus and Aedes aegypti were responsible for Filariasis and Dengue. Synthetic pesticides were effective against mosquitoes as well as main sources of environmental pollution and most of them are immunosuppressant. Botanicals were widely used as insecticides, growth disruptors, repellents, etc. The aim of this research was to determine larvicidal properties of powdered leaf, Elytraria acaulis against late third or early fourth in-star larvae of Cx. quinquefasciatus and Ae. aegypti. METHODS: Larvae of Cx. quinquefasciatus and Ae. aegypti were tested at various concentrations of 100, 120, 140, 160, 180 and 200mg/100ml and mortality was recorded after 24h. The LC50 values of the E. acaulis leaf powder were calculated by Probit analysis. RESULTS: The plant powder exhibited strong larvicidal activity against Cx. quinquefasciatus with LC50 value of 116.07mg/100ml against Ae. aegypti 124.25mg/100ml respectively. The result indicated that the plant powder of E. acaulis showed potential larvicidal activity against Cx. quinquefasciatus and Ae. aegypti. CONCLUSION: The overall findings of the present investigation suggested that the E. acaulis highly effective against Cx. quinquefasciatus and Ae. aegypti larvae. Elytraria acaulis may be used as an alternative to synthetic chemical pesticides for control of vectors to reduce vector borne diseases and did not harm to total environment.

A novel biopesticide PONNEEM to control human vector mosquitoes Anopheles stephensi L. and Culex quinquefasciatus Say.

Organophosphate pesticides are widely used in vector mosquito management and agricultural pest management. These chemicals enter into natural water bodies and soil and cause hazards to the environment. The objective of this study was to prepare a natural pesticide which will not harm the environment and yet control vector mosquitoes. PONNEEM, a novel biopesticide, patented and prepared from the oils of Azadirachta indica and Pongamia glabra, was tested against Anopheles stephensi and Culex quinquefasciatus. One hundred percent larvicidal and ovicidal activities were observed at 0.1-ppm concentration of PONNEEM against the two mosquito species under laboratory and sunlight conditions up to 12 months from the date of manufacture. Very high oviposition reduction of 26.46 and 32.16 % is also recorded. Reductions in α-esterase level (0.0818 ± 0.340 and 0.2188 ± 0.003), ß-esterase level (0.0866 ± 0.026 and 0.0398 ± 0.010 µg naphthol produced/min/mg larval protein), glutathione S-transferase enzyme (14.2571 ± 0.51 and 15.3326 ± 0.51 µmol/min/mg larval protein) and total protein levels (0.0390 ± 0.008 and 0.1975 ± 0.029 mg/individual larva in treated groups of A. stephensi and C. quinquefasciatus at 0.1-ppm concentration, respectively. The non-target organisms such as Gambusia affinis and Diplonychus indicus were not affected. Biopesticides are good alternatives to synthetic pesticides. PONNEEM can be effectively used for the management of human vector mosquitoes. Since it has a biodegradable nature and does not alter the environmental condition of water and soil.

Effect of confertifolin from Polygonum hydropiper L. against dengue vector mosquitoes Aedes aegypti L.

The essential oil from the leaves of Polygonum hydropiper L. (Polygonaceae) was tested against Aedes aegypti L. The LC50 values were 190.72 and 234.37 ppm against second and fourth instar larvae of A. aegypti, respectively. Confertifolin (6,6,9a-trimethy l-4,5,5a,6,7,8,9,9a-octahydronaphtho [1,2-c] furan-3 (1H)-one) was isolated from the essential oil of P. hydropiper leaves using silica gel column chromatography. The LC50 values were 2.90 and 2.96 ppm for second and fourth instar larvae of A. aegypti, respectively. At 10 ppm, the concentration of confertifolin showed ovicidal activity of 100, 100, and 77.6 % on 0-6, 6-12, and 12-18 h old eggs; the repellent activity was 323.2 min; and oviposition deterrent activity was 97.52 % and adulticidal activity was 100 % against A. aegypti. The results were statistically significant at P < 0.05 level. The results suggested that confertifolin as an effective major constituent against A. aegypti and might be considered as a potent source for the production of superior natural mosquitocides.

Effect of Polygonum hydropiper L. against dengue vector mosquito Aedes albopictus L.

The present study aimed to evaluate the essential oil and an isolated compound from the leaves of Polygonum hydropiper L. against dengue vector mosquito Aedes albopictus L. The plant material was macerated and steam distilled using clavenger apparatus for oil extraction. The essential oil was tested at different concentrations of 100, 50, 25, 12.5 and 6.25 ppm concentrations against the larvae of Ae. albopictus. The isolated compound was tested for larvicidal, ovicidal, repellent, oviposition deterrent and adulticidal activities at 10, 5, 2.5, 1.25 and 0.625 ppm concentrations. The essential oil exhibited LC50 values of 194.63 and 199.65 and confertifolin exhibited LC50 values of 2.02 and 3.16 against the second and fourth instar larvae of Ae. albopictus, respectively. The ovicidal activity of 100% on 0- to 6-h-old eggs, repellent activity of 320.6 min, oviposition deterrent activity of 98.51% and adulticidal activity of 100% at 10 ppm concentration of confertifolin were recorded. No mortality of was observed in negative control. To the best of our knowledge, this is the first report on the potential mosquitocidal activities of confertifolin against Ae. albopictus.

Bioefficacy of essential oil from Polygonum hydropiper L. against mosquitoes, Anopheles stephensi and Culex quinquefasciatus.

The biological activity of essential oil extracted from the leaves of Polygonum hydropiper and a compound, confertifolin, isolated from this plant was bioassayed against larva of mosquitoes, Anopheles stephensi and Culex quinquefasciatus. The essential oil showed the LC50 values of 189 and 243ppm; 217 and 242ppm, confertifolin (6,6,9a-trimethyl-4,5,5a,6,7,8,9,9a-octahydronaphtho[1,2-c]furan-3(1H)-one) showed the LC50 values of 2.40 and 3.09ppm; 4.07 and 4.18ppm against the second and fourth instar larvae of An. stephensi and Cx. quinquefasciatus, respectively. At 10ppm confertifolin showed ovicidal activity of 100, 98.6 and 86.4% against An. stephensi and 100, 100 and 75.2% against Cx. quinquefasciatus on 0-6, 6-12 and 12-18h old eggs; the repellent activity persisted for 314.6 and 319.0min; oviposition deterrent activity was 97.2 and 99% and adulticidal activity was 100 and 100% against An. stephensi and Cx. quinquefasciatus, respectively. The results were statistically significant. Confertifolin could be considered for use in the control of human vector mosquitoes.

A novel herbal formulation against dengue vector mosquitoes Aedes aegypti and Aedes albopictus.

The objective of this study was to develop a herbal formulation to control dengue vector mosquitoes. PONNEEM, a novel herbal formulation prepared using the oils of neem (Azadirachta indica), karanj (Pongamia glabra) and their extracts, was tested for larvicidal, ovicidal and oviposition deterrent activities against Aedes aegypti and Aedes albopictus at 1, 0.5, 0.3 and 0.1 ppm concentrations. Cent percent larvicidal and ovicidal activities were observed at 0.1 ppm in the two mosquito species under laboratory and sunlight-exposed conditions up to 12 months from the date of manufacture. Oviposition deterrent activity of 69.97% and 71.05% was observed at 1 ppm concentration of PONNEEM against A. aegypti and A. albopictus, respectively. Reduction in enzyme levels for α-esterase was 0.089 ± 0.008 and 0.099 ± 0.140 µg napthol produced/min/mg larval protein; for ß-esterase, it was 0.004 ± 0.009 and 0.001 ± 0.028 µg napthol produced/min/mg larval protein; for glutathione S-transferase, it was 10.4814 ± 0.23 and 11.4811 ± 0.21 µmol/min/mg larval protein and for total protein, it was 0.177 ± 0.010 and 0.008 ± 0.005 mg/individual larva in treated groups of A. aegypti and A. albopictus, respectively. The nontarget organisms such as Gambusia affinis and Diplonychus indicus were not affected. No mortality was observed in control. PONNEEM can be used effectively for the management of human vector mosquitoes.