Synthesis and larvicidal efficacy of pyrazolopyrimidine derivatives conjugated with selenium nanoparticles against Culex pipiens L. and Musca domestica L. larvae.

The synthesized pyrazolopyrimidine derivatives conjugated with selenium nanoparticles were prepared via a reaction of pyrazolone 1 with aryl-aldehyde and malononitrile or 3-oxo-3-phenylpropanenitrile in the presence ammonium acetate or pipridine using an ultrasonic bath as a modified method in the organic synthesis for such materials. The structure of the synthesized compounds was elucidated through various techniques. All the synthesized pyrazolopyrimidines were used in the synthesis of selenium nanoparticles (SeNPs). These nanoparticles were confirmed using UV-spectra, Dynamic Light scattering and (TEM) techniques. The larvicidal efficiency;of the synthesized;compounds; was investigated against some strains such as Culex pipiens;and Musca domestica larvae. Bioassay test showed pyrazolopyrimide derivatives to exhibit an acceptable larvicidal;bio-efficacy. The derivative (3) exhibited;the highest;efficiency for more than; lab strains of both species. Moreover, C. pipiens larvae were more sensitive towards the examined compounds than M. domestica. The field;strain displayed lower affinity for the 2 folds compounds. Some biochemical changes were tracked through analysis of insect main metabolites (protein, lipid and carbohydrate), in addition to measuring the changes in seven enzymes after treatment. Generally, there was a reduction in the protein, lipids and carbohydrates after treatment with all tested compounds. Moreover, a decrement was noticed for acetylcholine esterase and glutathione;S-transferase; enzymes. There was an increment in the acid;phosphatase; and alkaline phosphatase. In addition, there was elevation in Phenoloxidase level but it noticed the declination in both Cytochrome P450 and Ascorbate peroxidase activity after treatment both flies with derivatives of selenium-nanoparticles in both lab and field strain. Generally, the experiments carried out indicate that antioxidant and detoxification enzymes may play a significant role in mechanism of action of our novel nanocompounds. The cytotoxicity of the synthesized compounds and conjugated with SeNPs showed enhanced compatibility with human normal fibroblast cell line (BJ1) with no toxic effect.

Chemical composition of four essential oils and their adulticidal, repellence, and field oviposition deterrence activities against Culex pipiens L. (Diptera: Culicidae).

Effective mosquito repellents can limit the transmission of vector-borne diseases to humans. Consequently, there is an urgent need to develop mosquito control strategies that prioritize eco-friendly and cost-effective repellents. Essential oils (EOs) have enormous potential for mosquito repellency. Here, cinnamon, basil, eucalyptus, and peppermint EOs were investigated for adulticide and repellency properties against Culex pipiens as well on the oviposition behavior of gravid females from laboratory (lab test) and field (field test) populations. Cinnamon oil was an effective oviposition deterrent regardless of the population and had high adulticidal activity with toxicity index of 75.00% at 24 h of exposure, relative to deltamethrin. In addition, it exhibited effective repellency at 98.01% and 71.22% at 6.67 and 1.71 µl/cm2, respectively. Peppermint oil had the least adulticidal activity with toxicity index of 6.2% at 24 h, and it resulted in low repellency at 70.90% and 50.64% at 6.67 and 1.71 µl/cm2, respectively. On average, basil and eucalyptus oils showed some adulticidal efficiency, repellency, and oviposition deterrent activity. For all treatments, the oviposition deterrent index values of gravid females from natural populations (field test) were lower than those from lab-reared (lab test) females. Different ratios of monoterpenoids, phenylpropanoids, and fatty acids in the EOs tested likely account for the activity variations observed. Our results suggest cinnamon, basil, eucalyptus, and peppermint EOs, which are widely available, economical, and eco-friendly, with good potential for mosquito control strategies.

Anti-Culex pipiens activity of different pomegranate cultivars and determination of their bioactive compounds using LC-MS profiling.

INTRODUCTION: Pomegranate (Punica granatum L.) peels are rich in various bioactive compounds. Characterization of these compounds is crucial for the utilization of peel waste in industrial processing. OBJECTIVE: The study aimed (1) to establish and compare the metabolic profiles of the peel of seven pomegranate cultivars and (2) to identify bioactive compounds contributing to the larvicidal activity against the third instar larvae of Culex pipiens. MATERIALS AND METHODS: UPLC-ESI-MS/MS was utilized to analyze peel methanol extracts of different pomegranate cultivars. The larvicidal activity was determined by calculating the larval mortality among the third instar larvae of C. pipiens. Multivariate data analysis was conducted to identify the metabolites that exhibited a larvicidal effect. RESULTS: A total of 24 metabolites, including hydrolyzable tannins, flavonoids, and alkaloids, were tentatively identified in both negative and positive ionization modes. The extract of cultivar 'Black' exhibited the most potent larvicidal effect with LC50 values of 185.15, 156.84, and 138.12 ppm/mL after 24, 48, and 72 h of treatment, respectively. By applying chemometric techniques, the larvicidal activity could be directly correlated to the bioactive compounds punicalagin, quercetin-O-rhamnoside, quercetin-O-pentoside, and galloyl-HHDP-glucose. CONCLUSION: The present study implemented UPLC-ESI-MS/MS and chemometric techniques as potential tools for metabolomics analysis and differentiation between peels of different pomegranate cultivars. In addition, cultivar 'Black' extract could be a promising natural insecticide against mosquitoes since it is rich in bioactive compounds with larvicidal activity.

Human protozoa infection and dysplasia in ulcerative colitis: a neglected aspect in a prominent disease.

The chance of getting colorectal cancer (CRC) is higher in people with chronic ulcerative colitis (UC). The impact of parasitic infections on UC is underappreciated. The purpose of this study was to look into the effect of intestinal protozoal infections on the dysplastic changes generated by UC. The research included 152 adult patients with histologically confirmed UC and 152 healthy controls. Fecal samples were examined for the presence of parasites and fecal calprotectin (FC). The enzyme-linked immunosorbent assay measured serum anti-p53 antibodies (p53Abs) and metallothioneins (MTs). The advanced oxidation protein products (AOPPs) and reduced glutathione (GSH) levels were measured by a spectrophotometric method in all subjects. Serum C-reactive protein (CRP) and IL-6 were also measured. In addition, histopathological and immunohistochemical investigations of intestinal tissue were done. Our results exhibited significant increases in FC and CRP, IL-6, AOPPs, MTs, and p53Abs in ulcerative colitis patients with parasitic infections compared to those without parasites. In contrast, GSH levels showed a significant decrease in the same group compared with other groups. Histopathological and immunohistochemical assessments of intestinal tissue signified severe inflammation and strong expression of PD-L1 in patients with parasitic infections compared to others without parasitic infections. Our research indicated a greater frequency of intestinal protozoa in UC patients with elevated inflammatory and dysplastic biomarker levels. This suggests that these parasites may be involved in the etiology of chronic UC and the associated carcinogenetic process. This is the first report of a link between parasitic infections and dysplastic alterations in UC patients.

Co-application of entomopathogenic fungi with chemical insecticides against Culex pipiens.

Culex pipiens (Diptera: Culicidae) is a vector of many human and animal diseases. Its control is regarded as a preventative approach that is focused on effectively managing such diseases. In this context, dose response assays of two insecticides, bendiocarb and diflubenzuron were performed with two entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae against 3rd instar C. pipiens larvae. The most effective agents, combination experiments as well as enzymatic activities of phenoloxidase (PO) and chitinase (CHI) were also assessed. The results showed that diflubenzuron was more effective at low concentrations (LC50: 0.001 ppm) than bendiocarb (LC50: 0.174 ppm), whereas M. anisopliae was more effective (LC50: 5.2 × 105 conidia/mL) than B. bassiana (LC50: 7.5 × 107 conidia/mL). Synergistic interactions were observed when diflubenzuron was applied at 2- and 4-days post- exposure to M. anisopliae, with the highest degree of synergism observed when diflubenzuron was applied 2 days post-fungal exposure (χ2 = 5.77). In contrast, additive interactions were recorded with all other insecticide-fungal combinations. PO activities significantly (p ≤ 0.05) increased during 24 h after a single diflubenzuron treatment as well as when diflubenzuron was applied prior to M. anisopliae, whereas suppressed after 24 h when M. anisopliae applied prior to diflubenzuron as well as after 48 h from single and combined treatments. CHI activity increased 24 h after both single and combined treatments, the activity remained elevated 48 h after a single diflubenzuron treatment and when diflubenzuron was applied after M. anisopliae. Histological study of the cuticle by transmission electron microscopy revealed abnormalities following single and combined treatments. Germination of the conidia and production of the mycelium that colonizes the lysing cuticle was obvious when diflubenzuron was applied 48 h after M. anisopliae exposure. Overall, these results demonstrate that M. anisopliae is compatible with diflubenzuron at lower concentrations and that combined applications can improve C. pipiens management.

Phytochemical Investigation of Three Cystoseira Species and Their Larvicidal Activity Supported with In Silico Studies.

Culex pipiens mosquitoes are transmitters of many viruses and are associated with the transmission of many diseases, such as filariasis and avian malaria, that have a high rate of mortality. The current study draws attention to the larvicidal efficacy of three methanolic algal extracts, Cystoseira myrica, C. trinodis, and C. tamariscifolia, against the third larval instar of Cx. pipiens. The UPLC-ESI-MS analysis of three methanol fractions of algal samples led to the tentative characterization of twelve compounds with different percentages among the three samples belonging to phenolics and terpenoids. Probit analysis was used to calculate the lethal concentrations (LC50 and LC90). The highest level of toxicity was attained after treatment with C. myrica extract using a lethal concentration 50 (LC50) of 105.06 ppm, followed by C. trinodis (135.08 ppm), and the lowest level of toxicity was achieved by C. tamariscifolia (138.71 ppm) after 24 h. The elevation of glutathione-S-transferase (GST) and reduction of acetylcholine esterase (AChE) enzymes confirm the larvicidal activity of the three algal extracts. When compared to untreated larvae, all evaluated extracts revealed a significant reduction in protein, lipid, and carbohydrate contents, verifying their larvicidal effectiveness. To further support the observed activity, an in silico study for the identified compounds was carried out on the two tested enzymes. Results showed that the identified compounds and the tested enzymes had excellent binding affinities for each other. Overall, the current work suggests that the three algal extractions are a prospective source for the development of innovative, environmentally friendly larvicides.

Larvicidal and repellent potential of Ageratum houstonianum against Culex pipiens.

Mosquitoes are unquestionably the most medic arthropod vectors of disease. Culex pipiens, usually defined as a common house mosquito, is a well-known carrier of several virus diseases. Crude ethanol extracts of different organs of Agratum houstonianum are tested with Culex pipiens Linnaeus (Diptera: Culicidae) to determine their larvicidal, antifeedant, and repellency effects. Alongside biochemical analysis, the activity of the AChE, ATPase, CarE, and CYP-450 is detected in the total hemolymph of the C. pipiens larvae to examine the enzymatic action on the way to explain their neurotoxic effect and mode of action. Through HPLC and GC-MS analysis of the phytochemical profile of A. houstonianum aerial parts is identified. The larvicidal activity of aerial parts; flower (AF), leaf (AL), and stem (AS) of A. houstonianum extracts are evaluated against the 3rd instar larvae of C. pipiens at 24-, 48- and 72-post-treatment. A. houstonianium AF, AL, and AS extracts influenced the mortality of larvae with LC50 values 259.79, 266.85, and 306.86 ppm, respectively after 24 h of application. The potency of AF and AL extracts was 1.69- and 1.25-folds than that of AS extract, respectively. A high repellency percentage was obtained by AF extract 89.10% at a dose of 3.60 mg/cm2. A. houstonianium AF prevailed inhibition on acetylcholinesterase and decrease in carboxylesterase activity. Moreover, a significant increase in the ATPase levels and a decrease in cytochrome P-450 monooxegenase activity (- 36.60%) are detected. HPLC analysis prevailed chlorogenic and rosmarinic acid as the major phenolic acids in AL and AF, respectively. GC-MS analysis of A. houstonianum results in the identification of phytol as the major makeup. Precocene I and II were detected in AF. Linoleic, linolenic, and oleic acid were detected in comparable amounts in the studied organs. Overall, results suggest that the A. houstonianum flower extract (AF) exhibits significant repellent, antifeedant, and larvicidal activities.

Facile synthesis, larvicidal activity, biological effects, and molecular docking of sulfonamide-incorporating quaternary ammonium iodides as acetylcholinesterase inhibitors against Culex pipiens L.

Insecticides participate with a vital role in our lives especially in preventing the spread of human diseases via controlling the dangerous pests. It is a challenge to identify alternatives to the ordinary insecticides with new mode of action to be used for mosquitoes' control in an environmentally sustainable manner. Using a facile two-step procedure, three novel series of sulfonamide-incorporating quaternary ammonium iodides (3a-i, 4a-i and 5a-i) were synthesized and their chemical structures were successfully characterized. The uncharged sulfonamide intermediates (2a-i) were constructed through simple amidation of the corresponding (hetero)aryl sulfonyl chlorides then the cationic target molecules were formed by quaternizing the tertiary nitrogen with methyl, ethyl, and allyl iodides. The larvicidal activities and biological effects of most synthesized compounds against Culex pipiens L. were extensively investigated and they exhibited good and comparable activities to temephos. Among these hybrids, 4a showed the most potent activity with LC50 = 26.71 ppm. Additionally, the developmental durations of larval and pupal stages were significantly prolonged after treatment with all concentrations of 4h. At high concentration (160 ppm) of 4a and 4b, no adults emerged due to the complete death of pupae, and consequently zero growth index. Moreover, the results of the molecular docking demonstrated that the activities of compounds correlate partially to their binding with acetylcholinesterase (AChE) and it is not the sole parameter for determining the activity.

Screening and evaluation of different algal extracts and prospects for controlling the disease vector mosquito Culex pipiens L.

Continual application of synthetic insecticides in controlling mosquito larvae has resulted in several problems as build-up of mosquito resistance beside to negative impacts on human health and environment. Discovering new and affordable bio-insecticidal agents with high efficiency, cost effective and target specific become a crucial need. The current study assessed the larvicidal activity of eight methanolic algal extracts belong to three different algal divisions against the 3rd larval instar of Culex pipiens L. (Diptera: Culicidae). Comparative studies showed that four species of red and green algal extracts exhibited good larvicidal activity. Galaxaura elongata and Jania rubens (Rhodophyta), Codium tomentosum and Ulva intestinales (Chlorophyta) showed higher larvicidal potencies than Padina boryana, Dictyota dichotoma, and Sargassum dentifolium (Phaeophyta) and Gelidium latifolium (Rhodophyta). The maximum level of toxicity was achieved by exposure to G. elongata extract with LC50 (31.13 ppm), followed by C. tomentosum (69.85 ppm) then J. rubens (84.82 ppm) and U. intestinalis (97.54 ppm), while the lowest toxicity exhibited by G. latifolium (297.38 ppm) at 72 h post- treatment. The application of LC50 values of G. elongate, J. rubens, C. tomentosum, and U. intestinalis extracts affected the activities of antioxidant enzymes viz. superoxide dismutase, catalase and glutathione peroxidase as oxidative stress markers. An increase of antioxidant enzymes activities was recorded. Therefore, a significant elimination of free radicals, causing toxic effects. Overall, this study casts light on the insecticidal activity of some algal extracts, suggesting the possibility of application of these bio- agents as novel and cost- effective larvicides.

Virulence of entomopathogenic fungi against Culex pipiens: Impact on biomolecules availability and life table parameters.

Culex pipiens mosquitoes considered as vectors for many arboviruses such as the West Nile virus and encephalitis virus showing a global impact on human health. The natural management of the aquatic stages of this pest is crucial for maintaining an insecticide-free and sustained environment. The present work focused on studying the biological and biochemical effects of the entomopathogenic fungi: Metarhizium anisopliae, Beauveria bassiana, and Paecilomyces lilicanus, against 3rd instar larvae of Culex pipiens laboratory colony. The results revealed that M. anisopliae showed maximum larval mortality (88%) with the lowest lethal time (LT50) (22.6 hrs) at 108 spores/ml followed by B. bassiana (73.33%) with LT50 (38.35 hrs), while P. lilicanus showed minimum percent mortality (65%) with highest LT50 (51.5 hrs). The median lethal concentration (LC50) values were found to be 1.027 × 105 spores/ml for M. anisopliae, 1.24 × 106 spores/ml for B. bassiana, while it was 8.453 × 106 spores/ml for P. lilicanus. A reduction in female fecundity, number of hatched eggs, pupation and adult emergence percentage were recorded. The biochemical analysis of the treated larvae revealed different quantitative decrease in total soluble proteins, lipids, and carbohydrate hydrolyzing enzymes compared to control. Histopathological effects of fungal infection upon insect cuticles, muscles, and midgut were investigated. Based on the obtained results, M. anisopliae proved its superior virulent effect as a bio-control agent against Cx. pipiens.

New quinolizidine alkaloid and insecticidal activity of Sophora secundiflora and Sophora tomentosa against Culex pipiens (Diptera: Culicidae).

Phytochemical investigation of Sophora secundiflora alkaloid fraction led to isolation of one new quinolizidine alkaloid (1) 13-methoxyanagyrine together with six known ones (2-7). The insecticidal activity of 70% methanol extract of leaves of S. secundiflora, S. tomentosa and the isolated alkaloids were assessed against 3rd instar larvae of Culex pipiens (Diptera: Culicidae) using different concentrations and mortality rate was recorded. Sophora tomentosa extract showed highest mortality rate with median lethal concentration LC50 3.11 ppm after 24 h and 0.66 ppm after 48 h and anagyrine (6) exhibited remarkably insecticidal activity with LC50 value of 3.42 ppm after 24 h of exposure. Additionally, cytotoxic activity of alkaloid fraction of S. secundiflora, S. tomentosa and isolated alkaloids was also studied using crystal violet assay against MCF-7 and HEPG-2 cell lines. Anagyrine (6) exhibited IC50 values of 27.3 ± 0.7 and 30.2 ± 0.9 µg/mL against MCF-7 and HEPG-2 cancer cells, respectively.

Agro-waste derived compounds (flax and black seed peels): Toxicological effect against the West Nile virus vector, Culex pipiens L. with special reference to GC-MS analysis.

The development of different approaches to use agricultural residues as a source of high value-added products, become a must, especially after the problems emerged due to their accumulation. This contribution demonstrates the potential of agricultural residues, Linuim usitatissium (flax seed) and Nigella sativa (black seed) peels, as raw materials for the production of bioactive products, botanical insecticides, against Cx. pipiens, with deep analysis to their chemical constituents by gas chromatography-mass spectrometry, the larvicidal efficacies of the three crude extracts (methylene chloride, petroleum ether and methanol 70%) from the two plant waste peels were evaluated for the first time against the late third instar larvae of Cx. pipiens. Results indicated different lethal doses in larvae depending on the efficacy of organic solvent used. For both compounds methanol 70% extracts produced the highest dry yield. The most efficient solvent is petroleum ether in case of both flax and Black seed peels. Petroleum ether extract exhibited the highest toxicity against Cx. pipiens with an LC50 of 69.6383 ppm. The same results for black seed indicated that petroleum ether was the most efficient against Cx. pipiens with an LC50 of 40.7748 ppm. The study revealed for the first time the type of phytochemical constituents presents in peels of flax and black seeds using GC-MS analysis which revealed twenty-eight constituents among extracts of flax and black seed peels ranging from to 58.8711% to 99.99% of the total extracts. GC-MS profiling showed that a five constituents, 9-2-Methyl-Z, Z-3, 13 octadecadienol (terpenoid), 9,17-Octadecadienal, (Z)-, Nonanoic acid, 9-oxo-, methyl ester, 9,12-Octadecadienoic acid Z,Z and Octasiloxane, 1,1,3,3,5,5,7,7,9,9,11,11,13,13,15,15-hexadecamethyl- have insecticidal activity beside many other biological activities as recorded from a variety of botanical extracts. While the constituents like Hexadecanoic acid, methyl ester and cis-9-Hexadecenal, both of them are larvicidal, cis-Vaccenic acid and 9-Oxononanoic acid showing only an insecticidal activity beside Undecanoic acid the mosquito repellent. The other six constituents Linoelaidic acid, Oleic Acid, Z-2-Octadecen-1-ol, 1-Methoxy-3-hydroxymethylheptane, Cis-11,14-Eicosadieonic acid-methyl ester and Heptasiloxane, 1,1,3,3,5,5,7,7,9,9,11,11,13,13-tetradecamethyl- are constituents of other plant extracts which showed as a whole an insecticidal activity.

The larvicidal activity of natural inspired piperine-based dienehydrazides against Culex pipiens.

A series of piperine-based dienehydrazide derivatives were designed and synthesized to be used as insecticides against Culex pipiens. The chemical structure of compound 5n was confirmed by single-crystal x-ray diffraction. Their insecticidal activities of synthesized compounds were tested against third-instar larval of Cx. pipiens at concentrations ranging from 0.1 to 1.2 mg/mL. Among all derivatives, compounds 5a, 5b, 5f, 5g, 5m, 5n, 5o, 5p, and 5u displayed good activities. The final mortality rates at the concentration of 0.75 mg/mL after 48 h treatment, were found to be in the range from 80.00 to 83.33% and with LC50 values ranging from 0.221 to 0.094 mg/mL. These compounds demonstrated higher insecticidal activities than piperine and Deltamethrin (a commercial positive control). Molecular modelling reveals several molecular interactions between synthesized compounds and the substrate binding sits of acetylcholinesterase (AChE) that are predicted to be responsible for its binding and inhibition activity. .

Deroplax silphoides (Thunberg, 1783) (Hemiptera: Heteroptera: Scutelleridae) Invasive Species in Egypt with additional morphological and behavioral data.

Invasive species continue to be a noteworthy risk to a wide range of biological systems and species diversity throughout the world. At present, accessible data about invasive species in Egypt are still insufficient despite the genuine danger these species pose to the Egyptian environment, economy and human well-being. The present study adds new distribution records of the invasive scutellerid species, Deroplax silphoides (Thunberg, 1783), to the Egyptian fauna. In 2008, this bug was first observed in Egypt in South Sinai, followed by Giza in 2016. Recently, hundreds of specimens were reported from Cairo in 2018. A new host plant, Chrysanthemum morifolium L. (Asteraceae), is reported for this bug. This work also provides additional information about the male and female genitalia. New characteristic features, photos, examined material, map of its world distribution, ecological and behavioral notes are also provided. Moreover, egg sculpturing of Deroplax silphoides was examined for the first time.