Efficacy of Acacia nilotica, Eucalyptus camaldulensis, and Salix safsafs on the mortality and development of two vector-borne mosquito species, Culex pipiens and Aedes aegypti, in the laboratory and field

Mosquitoes are one of the most lethal animals in the world and transmit many dangerous human pathogens, causing millions of deaths each year. The search for modern and better mosquito control is an endless effort almost all over the world. Phytochemicals are promising biological agents for getting rid of pests that are harmful to human and animal health and crops, they are inexpensive, biodegradable, and have diverse modes of action. The efficacy of acetone and hexane leaf extracts of Acacia nilotica, Eucalyptus camaldulensis, and Salix safsafs was investigated against the 2nd and 4th larvae and pupae of two vectors, Culex pipiens and Aedes aegypti. The results showed the obvious effect of A. nilotica extract on the mortality of mosquito larvae, the reduction of female eggs, and a higher mortality rate in sunlight than in shadow (fluorescein). Data from field trials revealed that A. nilotica extracts had the greatest effect on larval reduction, reaching 89.8% in 24 h and having a 12-day stability. Polyethylene glycol, sesquiterpenes, and fatty acids were the most common compounds found in A. nilotica, E. camaldulensis, and S. safsafs, respectively. The acacia plant had promising larvicidal activity, safe and effective alternative to chemical insecticides.


Introduction
Plants contain phytochemicals, or secondary metabolics, and ingesting them typically has positive health impacts. Due to their positive impacts on human health and significant health benefits for consumers, phytochemicals are of great interest and have significant antioxidant potential [1][2][3]. According to preclinical, clinical, and epidemiological research, phytochemicals may be useful in treating a variety of ailments because of their antioxidant and anti-inflammatory properties. On the other hand, consuming some phytochemicals may have some short-term and long-term harmful effects, including the potential to cause cancer and many other diseases [4].
Aromatic medicinal plants have been used to treat many diseases in humans and animals, both in the medical field and in industry. As a result, consumers who care about the environment are using more natural insecticides in many parts of their lives [5,6].
Phytochemicals still contain many natural, organic compounds that have a toxic or repellent effect on many insect pests, which is what we aim to achieve and explore through this work. Therefore, the trees chosen are available in the Egyptian environment, in addition to the fact that it contains various toxic substances. The Acacia tree needs a lot of attention, whereas this tree has been used since the Pharaonic time because it (Acacia or Sunt) contains substances with unique properties [42]. These secondary metabolites are crucial in the interactions between plants and insects and are what give plants their insect resistance [30].
This study aimed to evaluate the larvicidal effectiveness of plant leaf extracts, Acacia nilotica, Eucalyptus camaldulensis, and Salix safsafs against 2nd, 4th larvae and pupae of Culex pipiens and Aedes aegypti besides determining some biological aspects after being treated with sub-lethal concentrations of plant extracts. The structure chart of the article is shown in Fig. 1.

Mosquito colony
Mosquito larvae, Aedes aegypti and Culex pipiens used for all investigations were obtained from Department of Entomology, Faculty of Science, Benha University. Mosquito larvae were reared in enamel plates (25 × 20 × 10 cm) filled with 2 L de-chlorinated tap water and given Tetramin® fish food and powdered dog biscuits every 2 day. At 27 ± 2 • C, 75-80% RH, and a 12:12 h (L/D) photoperiod, the colony was kept in good condition. After pupation, Ae. aegypti was transferred into mosquito cage (30 × 25 × 25 cm) and Cx. pipiens pupae into (30 × 30 × 30 cm) cages in an insectary. Mosquito adults used 8-10% sucrose solution as food. Egg-laying dishes for Ae. aegypti mosquitoes are provided with a paper or a thin foam layer that sticks to the edge of the egg-laying dish to help females stand on it while laying eggs, in addition to facilitating the collection of eggs through that sheet. Both larval species were kept in the identical laboratory settings and were continually available for the tests [43].

Collection of plant materials
Leaves of Acacia nilotica (Fabaceae), Eucalyptus camaldulensis (Myrtaceae), and Salix safsafs (Salicaceae) were collected from different areas in Egyptian villages, Giza area, Egypt in May-September 2021 (Fig. 2). Plants were identified at Flora and Phytotaxonomic section, Agricultural Research Center, Giza, Egypt.

Preparation of plant extracts
Fresh leaves of A. nilotica, E. camaldulensis, and S. safsafs were washed with tap water and air dried. Stock solutions of plant leaves were extracted by mechanically grinding 50 g using a stainless-steel electric mixer and placing the powder in a Soxhlet apparatus for 4-7 h according to the type of solvent (acetone and hexane). The solution was filtered using Whatman No. 1 filter paper and dried in an oven at 30 • C for 6 h. The extracts were stored in a dark bottle in a refrigerator at − 5 • C for 24 h prior to the experiment.

Larvicidal and pupicidal efficacy
Plant extracts were tested for their larvicidal and pupicidal efficacy according to WHO [44]. against 2nd, 4th larval instar and pupal stage of Cx. pipiens and Ae. aegypti. Different concentrations were prepared as 62.5, 125, 250, 500, 1000, and 1500 ppm with twenty larvae were placed in a glass beaker containing 250 ml for two species. The experiment as well as the control group were treated with the solvent only. Five replicates were used for each concentration. Cx. pipiens and Ae. aegypti larval and pupal mortalities were recorded after 24 and 48 h post-treatment (PT).

Effect of plant extracts on larval development
The effect of sublethal (LC 50 : lethal concentrations of cumulative effect) of the plant extracts, A. nilotica, E. camaldulensis, and S. safsafs was tested through LC 50 values which were determined with the test larvicide after 48 h post-treatment with plant extracts. There were five replicates, each containing a total of 30 Cx. pipiens and Ae. aegypti from the 3rd larval instar. Daily readings were performed to check for larval stage, behavioral changes, the presence of exuvia, the emergence of adults, and potential mortality of larvae, pupae, and adults. The experiment was conducted until the death of the last pupae, or the last adult completely emerged at room temperature (27 ± 2 • C).

Effect of plant extracts on egg laying and hatching percentages
Testing of the efficacy of plant extracts on female egg-laying activity was performed using 20 females (3-5 days old) from both Cx. pipiens and Ae. aegypti after treating their larvae with their LC 50 and LC 90 values and monitoring egg laying in a bioassay cage (30 cm × 30 cm x 30 cm) containing cups with 350 ml of dis-tilled water for egg laying. Adults were given a steady supply of an 8% glucose solution. A piece of filter paper (Whatman® No. 1) was placed on the inner surface of each plastic cup to act as a support for female Ae. aegypti eggs. Under a stereomicroscope, the number of eggs laid in the cups for each treatment group was recorded.

Larvicidal activity of plant extracts as photosensitizer
The effect of plant extracts (A. nilotica, E. camaldulensis, and S. safsafs) was tested against Cx. pipiens and Ae. aegypti through a larval dipping test, according to a previously described protocol [44], after 24 h of post-treatment to determine the efficacy of plant extracts as photosensitizers. Two concentrations of plant extracts (62.5 and 125 ppm) were freshly prepared in distilled water. There were five replicates, each containing a total of 100 Cx. pipiens and Ae. aegypti from the 4th larval instar.

Phytochemicals analysis
Thermo Scientific Trace GC Ultra/ISQ Single Quadrupole MS, TG-5MS fused silica capillary column, 0.1 mm, 0.251 mm, and 30 m thick were utilized for the GC/MS was employed for the biochemical analyses. It was achieved using an electronic ionizer with a 70 eV ionization energy. As a carrier gas, helium gas was used (flow rate: 1 ml/min). The MS transmission line and injector were both set to 280 • C. The oven was preheated to 50 • C, then increased to 150 • C at a rate of 7 • C per minute, 270 • C at a rate of 5 • C per minute (pause for 2 min), and lastly 310 • C at a rate of 3.5 • C per min (continued for 10 min). A relative peak area was employed to explore the quantification of all components discovered. The chemicals were at least partially identified by comparing the retention times and mass spectra of the chemicals to those of NIST, Willy Library data from the GC/MS instrument. Identification was done using the aggregate spectrum of user-generated reference libraries. To evaluate peak homogeneity, single-ion chromatographic reconstructions were performed. To verify GC retention periods, co-chromatographic analysis of reference substances was used whenever practical [45].

Larvicidal field assessment
In Kafr Saad village, Qalyubiya Governorate, Egypt, pools were selected (with average lengths of 6.5 m, 2.0 m, and 0.45 m) for the assessment of plant extracts. The pools used for larval breeding were chosen because they had a high density of mosquito stages. Each plant extract was applied in the breeding areas with a dose of LC95 × 2. Three replicates of each treatment were carried out. mosquito larvae were collected in field water from each site using an enamel pad (450 mL) in each larvicidal treatment [46].

Data analyses
The data were analyzed by the software, SPSS V23 (IBM, USA), for doing the Probit analyses to calculate the lethal concentration (LC) values and the one-way analysis of variance (ANOVA) (Post Hoc/Turkey's HSD test). The significant levels were set at p < 0.05.

Larvicidal activity
This work evaluated three plant extracts of Acacia nilotica, Eucalyptus camaldulensis, and Salix safsafs against the 2nd, 4th larval instars, and pupae of Culex pipiens and Aedes aegypti, and expressed dose and time-dependent efficacy extended at 2 days posttreatment.
The mortality % 24 h post-treatment using 1500 ppm, acetone extracts of Acacia nilotica, Eucalyptus camaldulensis, and Salix safsafs were 100% for 2nd, 100, 95, and 100% for 4th larval instar and 82, 70, and 77% for pupae of Cx. pipiens with LC 50 (50%, median lethal Unexpected, the morality recorded in Eucalyptus camaldulensis hexane extract was effective than acetone, where the morality reached 100, 96, and 54% for 2nd, 4th larval instars, and pupal stage respectively. In acetone extract, the mortality reached 100, 90, and 60% at 1000 ppm, 48 h post-treatment ( The results showed that all plant extracts in this study showed moderate to high toxicity effects against mosquito species Cx. pipiens and Ae. aegypti, and acetone extracts were more effective than hexane extracts.  Table 7). The results also showed that those effects were obvious on Ae. aegypti larvae, where it took 16, 14.7, and 18.5 days for the larvae to reach the adults in acetone extracts, and 13.6, 14.5, and 16.8 days in hexane extracts, respectively. The cumulative larvicidal mortality rates were 92.2, 81.1, 80, and 80, 82.2 and 75.6%, respectively (Table 7). For E. camaldulensis extract, it had the least effect on the elongation of mosquito larval growth * LC50 values = lethal concentration that kills 50% of the exposed larvae; (95%CL) = lower and upper confidence limit; ** Regression line equation; X2 = chi-square; Significant at P < 0.05 level.

Effect of sub-lethal on larval development
in Cx. pipiens or Ae. aegypti larvae, where the experiment took 10.5 and 14.7 days to reach adult with a mortality rate of 75.6 and 81.1%, respectively. During the experiments, it was observed that the larvae were able to develop and reach the adult stage. The plant extracts that significantly reduced the adult emergence rate of Cx. pipiens and Ae. aegypti was A. nilotica plant, either in acetone (5.6 and 8.9 days) or hexane (12.2 and 15.6 days) extracts (Table 7).

Oviposition bioassay
To evaluate the effectiveness of the effect of A. nilotica, E. camaldulensis, and S. safsafs plant extracts on the reduction of egg laying and hatching percentages in Cx. pipiens and Ae. aegypti females at LC 50 and LC 90 concentrations, the results showed a clear effect of the plant extracts in reducing the rate of egg laying by female mosquitoes at the selected concentrations. The average number of eggs laid by female Cx. pipiens in cups was 28.5, 72, and 80.7 eggs at the LC 50 value and 8.1, 34.7, and 45.5 eggs at the LC 90 value. Similarly, Ae. aegypti female mosquitoes laid fewer eggs when exposed to LC 50 or LC 90 concentrations. The results also showed that the rate of   Table 8). The paired t-test revealed that these outcomes were statistically significant (P > 0.05).

Larvicidal activity of plant extracts as photosensitizer
All plant extracts in this study showed normal effects against Cx. pipiens and Ae. aegypti larvae after 24 h post-treatment, as shown in the larvicidal activity Tables (1-2, 5-6) at the same concentration. A. nilotica plant extracts showed high toxic effects on mosquito larvae, where the mortality % reached 36% and 93% at 62.5 and 125 ppm for Cx. pipiens in acetone, and 28% and 80% ppm in hexane extracts after sunlight exposure. In shadow light, the mortality % reached 13%, 34%, 8%, and 21% for A. nilotica acetone and hexane at 62.5 and 125 ppm, respectively (Table 9). Similarly, Ae. aegypti larvae mortality rates were 27% and 79% in sunlight for acetone and 22% and 70% for hexane, respectively, while in shadow light the mortality rates were 11 and 25% for acetone and 6 and 18% for hexane extracts (Table 9). Data showed that, at 125 ppm, the effects of E. camaldulensis and S. safsafs plant extracts were not affected on larval mortality either in sunlight or under shadow lights.

Discussion
Several plants with bioactive compounds have been used with great efficacy to control and reduce a variety of medical insects and crop pests. Plants like Pyrethrum species, Curcuma longa, Juniperus communis, and sweet wormwood, Artemisia annua, are examples of plants that have been effectively used as safe pesticide sources for the management of pests and malaria vector-borne diseases [47].
Before technology, the practice of pest management using plant products was the predominant and most widely used practice, but over time, technology took hold of pesticides and synthetic pesticides were developed [48]. Due to their success in controlling typhoid fever and malaria, along with important crop diseases such as rust and blights [49,50], synthetic pesticides were more quickly accepted than plant pesticides. As a result, the use of natural products of plant origin gradually decreased until recently when the use of synthetic pesticides began to endanger public health and the environment [8,51], and monitoring of hazardous chemical pesticide residues in foods [52].
Because of this, the world has trouble with pest control and control programs, even though people keep finding new biologically * LC50 values = lethal concentration that kills 50% of the exposed larvae; (95%CL) = lower and upper confidence limit; ** Regression line equation; X2 = chi-square; Significant at P < 0.05 level.
active products that are widely used to fight medical, veterinary, and agricultural pests.
Our results revealed that all plant extracts in this study showed moderate to high toxicity effects against Cx. pipiens and Ae. aegypti larvae after 24 and 48 h of exposure and acetone extracts were more effective than hexane extracts.
Among the plant extracts, A. nilotica was the most effective regarding lethal concentrations (LC 50 ) and had lower values compared to other plant extracts either in methanol or hexane extracts, at 24 and 48 h post-treatment. Parallel studies of using botanicals against Culex larvae were also recorded. The aqueous extract of A. nilotica was effectively controlled on mosquito Culex larvae at 2% concentration with mortality rate between 75% and 100% (LC 50 = 400 ppm (0.004%) [53].
Acacia nilotica seeds and leaves have reportedly been found to have a variety of multipurpose active compounds [27,54]. As a result, extracts from the leaves and fruits of A. nilotica have shown promise as fungicides, bactericides, molluscicides, and insecticides [27,55].
Acacia nilotica seed essential oil and seed pod solvent extracts for bioefficacy against three important types of mosquitoes were  While the values of smoke toxicities were 82% in Cx. quinquefasciatus, 90% in Ae. aegypti, and 80% mortality in An. stephensi adults [55]. Our data agreed with the work of Zaitoun et al. [56], who studied the effect of A. nilotica extracts on the larval Cx. pipiens. They found that A. nilotica acetone leaf extracts were acutely toxic at 212.1 mg/L and chronically toxic at 144.2 mg/L; in addition, acetone and hexane extracts have demonstrated the capacity to inhibit egg hatchability and adult emergence.
The extracts of the Acacia plant are characterized by various phytochemicals, including terpenes, unsaturated and monounsaturated fatty acids, carboxylic acid, alkanes, and other plant extracts. Terpenoids, terpenes, and carbonyl were found to be the most effective phytochemical insecticides when tested against beetles, mosquitoes, caterpillars larvae and other flies over a 76-year period of literature review and meta-analyses of phytochemical insecticides [57].
Squalene is a multipotent triterpenoid widely present in a variety of plants, as in Acacia nilotica. Many studies have reported the pharmacological efficacies of squalene, including its antioxidant, antimicrobial, anticarcinogenic, and bioinsecticide properties [58,59].
The use of plant secondary metabolites and essential oils against mosquito larvae and adults has been documented in several laboratory studies [60,61]. Acacia nilotica crude extracts have been shown to be toxic to larvae of many common mosquito species [27,56].
Besides the efficacy of A. nilotica on mosquito larvae, the methanol and acetone bark extracts of A. nilotica were tested for decreased  * The significant differences between the treated and untreated group were determined by paired t-test (P < 0.05).
growth of 1st, 2nd, and 3rd instar Bactrocera cucurbitae larvae using an artificial diet bioassay [63]. The authors showed that both extracts had a negative impact on B. cucurbitae, a dangerous pest of cucurbit crops, during both their larval stage and overall developmental span. While significantly inhibited pupation and emergence percentages were recorded [63]. The literature review reveals that there have been very few investigations on the effects of willow leaf extracts on mosquito larvae. A study by Sameeh et al. [64] showed that the LC 50 value of an ethanol extract of Salix willow leaves against An. pharoensis larvae were 73.1 ppm, which is noticeable when combined with the 24 h (LC 50 = 2100 ppm) against 4th instar larvae of Cx. pipiens [65].
Salix safsaf, the deciduous herb, cape silver willow, or safsaf willow, is another name for the little tree known as the Egyptian willow, which has been found in Egypt since prehistoric times [38,39]. Typically, it can be found in moist places like those near waterways and on the Nile River in Egypt. Salicin willow, another name for white willow, has long been used for its therapeutic properties [40]. Its branches are supple, long, and thin. Over many generations, the plant's leaves, seeds, and other parts have been Five replicates were used for each concentration (20 larvae/replicate were used).

Table 10
The major chemical constituents of Acacia nilotica acetone extracts. used in traditional medicine to relieve inflammation, pain, and fever [41].
Besides the evaluation of extracts of the willow tree (Salix safsaf) on mosquitoes is very rare, in addition to extracting the compound acetylsalicylic acid (Aspirin) from willow tree [66], which has multiple medical uses [67].
13 plant extracts were evaluated against Musca domestica. The prickly pear (Opuntia vulgaris) and sugarcane tree (Saccharum spp.) were excluded from the preliminary toxicity against M. domestica adult at 300 and 1000 ppm because they displayed very low toxicity even at the higher dose. According to their effectiveness, the bioassayed extracts could be grouped as follows: Salix safsaf (0.24 mg/ cm 2 ), Conyza aegyptiaca (0.25 mg/cm 2 ), Azadirachta indica (0.28 mg/cm 2 ) is followed by five extracts with the same RC 50 value (Recovery concentration) was 0.29 mg/cm 2 and (Sonchus oleracues, Citrus aurantifolia, Cichorium intybus, Zea mays and Piper nigrum) [68].
Salix safsaf leaf extracts were investigated for insecticidal efficacy, growth regulation, adult performance, and repellency against third-instar larvae of the housefly Musca domestica by Hasaballah et al. [69]. Data obtained showed that the percentage of deaths among pupae and third-instar larvae rose with concentration, where the LC 50  Previous studies on the genus Salix's phytochemical composition have produced findings on phenolic substances, flavonoids, terpenes, and lignans [70,71]. Salicylic glycosides, the most prevalent phenolic compounds found, have been shown to have analgesic, antipyretic, anti-inflammatory, and anti-rheumatic, antioxidant and anticancer activities [71,72].
In addition to the GC-MS profile of the S. safsafs acetone and hexane extracts, which showed that willow contain main chemical compounds were α-Amyrin (20.38%), and Lupeol (13.32%) that are triterpene, and 1-Heptacosanol (12.15%) and n-Hexadecanoic acid (9.91%) are fatty acids in S. safsafs acetone extracts. 1-Heptacosanol (29.51%) is an alkane and Hexacosanal (25.42%) is saturated fatty alcohol, while α-Sitosterol (13.86%) and Ethyl iso-allocholate (10.07%) are triterpene for S. safsafs hexane extracts, respectively, which are considered essential compounds. Secondary metabolites are a diverse class of molecules that plants manufacture to protect them from herbivores and microbes. Alkaloids, phenolics, and terpenoids make up a large portion of these secondary metabolites and these compounds are toxic substances against insects [76].
Pharmacological studies are closely related to phytochemical compounds. The goal of these studies is to find out how to use many of these phytochemical compounds for different medical, therapeutic, and cosmetic purposes. These phytochemicals, derived or extracted from several aromatic plants such as Malva sylvestris, A. nilotica, Ageratina pichinchensis, E. camaldulensis, Capparis spinosa, and many other aromatic plants, are responsible for many pharmacological activities such as anti-inflammatory, antimicrobial, hepatoprotective, laxative, antiproliferative, and antioxidant [77][78][79][80][81]. Similar studies indicated that methanol leave extract of A. nilotica contained calycanthidine, linoleic acid, catechine, malic acid and octadeconic acid [82]. The GC-MS profile of seed essential oil from A. nilotica showed the presence of hexadecane (18.440%) and heptacosane (15.914%), which are the main and active compounds, and which may be involved in insecticidal activity [83].
The antioxidant activity of nonvolatile chemicals in leaves of Eucalyptus camaldulensis trees was investigated, where the extracts obtained by ethanol digestion and supercritical fluid extraction (SFE; CO 2 with 15% ethanol) showed a new promising antioxidative activities [84].
Eucalyptus camaldulensis leaves extracts showed the gallic and ellagic acid were found to be the prevailing antioxidants in the ethanolic extract and the main two compounds; 5-hydroxy-7,49-dimethoxy flavone and 5-hydroxy-7,49-dimethoxy-8-methyl flavone of the supercritical fluid extraction extract with antioxidative activity revealed to be flavones [84,85].
The fruit volatile oil of E. camaldulensis var. brevirostris was extracted by hydrodistillation and ethanol, where the gas chromatography-mass spectrometry was used to analyses the volatile oil that was produced. There were 38 volatile components found. Aromadendrene (17.99%), α-pinene (12.68%), cubenol (9.23%), α-gurjunene (6.65%), p-cymenene (5.39%), thymol (1.62%), and pcymen-7-ol (0.73%) were the primary volatile components in the fruit volatile oil. Monoterpenes (20.6%), sesquiterpenes (33.8%), light-oxygenated (8.1%), and heavily oxygenated (37.6%) compounds were the four groups into which the volatile components were divided [86]. Our data showed that the extracts of A. nilotica are characterized by an amber color, and with the passing of time, they change to a dark brown color and then black, so the plant extracts were evaluated against mosquito larvae, Cx. pipiens and Ae. aegypti at diluted concentrations (62.5 and 125 ppm) in sunlight and shadow light. The biochemical analysis with the GC-MS chromatogram confirmed that the A. nilotica extracts contains a photosensitizer (Benz[e]acephenanthrylen-12-ol), which explains the highly toxic effects on mosquito larvae, where the mortality rates in sunlight exposure reached 93% and 80% at acetone and hexane extracts against Cx. pipiens, 125 ppm, respectively. While in the shadow, the mortality rates % reached 34, and 21%, respectively. Similarly, in sunlight, the mortality rates were 79% and 70% in acetone and hexane extracts, respectively against Ae. aegypti larvae, while in shadow light, the mortality rates were 25% and 18%, respectively. * Fatty acid methyl esters (FAME); aromatic carboxylic acid (ACA); sesquiterpene lactone (STL); saturated fatty acids (SFA).
The data of the present work confirmed that the extract of A. nilotica contains naturally occurring photosensitizing substances such as Fluoromethacin, dl-α-Tocopherol and Benz[E]acephenanthrylen-12-OL.
Natural photosensitizing compounds are considered safe mechanisms in the control of insect pests, in addition to their use in photodynamic therapy, which is a minimally invasive, alternative, and promising treatment for several diseases, including cancer, actinic keratosis, atherosclerotic plaques, and macular degeneration and other diseases [88]. * Fatty acid methyl ester (FAME); saturated fatty alcohol (SFA); unsaturated fatty acids (USFA). Insects have a significant negative impact on agriculture all over the world other than transmitting many deadly diseases to humans or animals. Crops have created defenses against these pests, including a variety of specific compounds based on secondary metabolites. These substances (phytoalexins, phytocytibins) can either directly affect pests or can be used unintentionally to attract their natural enemies [89].

Conclusions
The diseases that mosquitoes spread around the world must be prevented from fatally affecting humans and their animals. The basis of pest control is the use of traditional pesticides, yet virtually all pesticides are no longer effective against insects. Due to the great variety and high efficacy of phytochemicals compounds, the use of natural products as ecologically benign pesticides is currently a significant topic of research. Therefore, this study revealed for the first time the efficacy of acetone and hexane extracts of A. nilotica, E. camaldulensis, and S. safsafs against larvae and pupae of Culex pipiens and Aedes aegypti. Acetone plant extracts were more recommended than hexane extracts as an ideal eco-friendly and inexpensive pest control approach that could be incorporated into integrated pest management used for protecting human from vector-borne diseases. All plant extracts in this study showed moderate to high toxic effects against Cx. pipiens and Ae. aegypti, beside that A. nilotica was the most effective regarding lethal concentration, while willow extracts showed a clear effect on the delay of larval development. A. nilotica had a higher mortality rate in sunlight than in shade, either from acetone or hexane extracts because they contained the photosensitizer. A. nilotica had a higher mortality rate in sunlight than in shadow light, either from acetone or hexane extracts because they contained the photosensitizer (Benz [E] cephenanthrylen-12-ol) (1.03%), and this distinguishes the amber color of A. nilotica extracts that are with the passage of time it turns to dark brown and then black. General, local, regional, and rural populations with few other control options can manage mosquito vectors by using such eco-friendly, inexpensive plant extracts in safe phytochemical pesticides. Further studies could be directed towards studying the phytochemical compounds and field application, and the safety profile of Acacia nilotica, Eucalyptus camaldulensis, and Salix safsafs against non-target organisms.

Author contribution statement
Rowida S. Baeshen: Conceived and designed the experiments; Analyzed and interpreted the data; Contributed reagents, materials, analysis tools or data; Wrote the paper.
Mohamed M. Baz: Conceived and designed the experiments; Performed the experiments; Analyzed and interpreted the data; Contributed reagents, materials, analysis tools or data; Wrote the paper.

Data availability statement
Data will be made available on request. Funding: This research received no external funding Fig. 6. Field efficacy of A. nilotica (A), E. camaldulensis (B), and S. safsafs (C) treated at dose of LC 90 X2 (529.9, 1503.5 and 902.9 ppm), respectively, in larval breeding sites.