Water Extracts From Trees Negatively Affect Oviposition Behavior and Fitness of Southern House Mosquito, Culex Quinquefasciatus Say (Diptera: Culicidae): A Simple, Easily Adoptable and Cost-Effective Approach to Control Mosquitoes

Water Extracts From Trees Negatively Affect Oviposition Behavior and Fitness of Southern House Mosquito, Culex Quinquefasciatus Say (Diptera: Culicidae): A Simple, Easily Adoptable and Cost-Effective Approach to Control Mosquitoes | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Water Extracts From Trees Negatively Affect Oviposition Behavior and Fitness of Southern House Mosquito, Culex Quinquefasciatus Say (Diptera: Culicidae): A Simple, Easily Adoptable and Cost-Effective Approach to Control Mosquitoes Muhammad Khan, Rizwan shah, Sarfraz Ali Shad, Shahid Majeed, Shahbaz Ahmad, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-143809/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background: The Culex quinquefasciatus Say is an important vector of many diseases of public health and veterinary importance. Environmental pollution, resistance development and health hazards associated with the use of synthetic insecticides have led to explore eco-friendly and cost effective management methods for mosquitoes. Methods: In our study, simple water extracts from leaves and bark of three tree species; Eucalyptus camaldulensis, Azadirachta indica (Neem) , and Moringa oleifera, were tested to explore their potential negative effects on the oviposition behavior and fitness of Cx. quinquefasciatus . Results: Oviposition bioassays showed a significant delay in onset of 1 st oviposition in the all treatments as compred to control. Number of egg rafts and fecundity were significantly lower in all treatments compared to control. Larval emergence was significantly less in the all treatments as compared with the control except Moringa bark. Highest pupation was observed in control as compared to all other treatments. Importantly, no adult emerged in Eucalyptus leaf, bark and in Neem bark extracts. In case of toxicity bioassays, median lethal time (LT 50 ) was lower at higher concentrations and higher at lower concentrations. At highest tested concentration of every botanical extract, LT 50 was highly age dependent that is higher for elder larvae and vice versa. Conclusions: Therefore, it could be concluded that simple water extrcats, made locally, from trees may have a great potential to be incorporated in the mosquito control programs. Translational Medicine Botanical Extract Mosquito Management Oviposition Behaviour Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Background Southern house mosquito, Culex quinquefasciatus Say, is a vector of many zoonotic diseases that affect humans and wild and domestic animals, such as avian malaria, Western equine encephalitis, lymphatic filariasis, St. Louis encephalitis, West Nile fever and Zika virus (Guedes et al. 2017; Simonsen and Mwakitalu 2013). Mortality rate due to vector-borne filariasis is low but a high morbidity rate is a social stigma. According to estimates, about 120 million people in 81 countries are currently infected, and 1.34 billion people who live in areas endemic to filariasis are at risk of infection (Sangshetti et al. 2017). In Pakistan, population of Cx. quinquefasciatus develops rapidly during rainy season and becomes a sever biting nuisance leading to restless nights (Tahir et al. 2009). Although, the ornithophilic behavior of Cx. quinquefasciatus is well reported, but the preference changes to mammals in the absence of primary host, which leads to significant impact (Kilpatrick et al. 2006). Application of synthetic insecticides as larvicides, insecticide-treated nets and indoor residual spraying (IRS) are the principal mosquito management measures adapted worldwide (Okumu et al. 2011). So far, chemical insecticides are the most effective control measures against mosquitoes, however, resistance against these chemical insecticides has recently been reported in many species of mosquitoes, which is resulting in repeated epidemic (Guessan et al. 2007; Van Bortel et al. 2008). Moreover, residual effects of synthetic insecticides are imposing serious threat to humans, animals, plants health and environment (References?). These major concerns have diverted the attention of researchers towards natural chemicals, such as plant extracts or oils, which are cost effective, safe, target specific and easily biodegradable in the environment (Borah et al. 2010; Hafeez et al. 2011). It has been shown by several studies that plant extracts or their non-volatile and volatile constituents have a great potential to be used as insecticides and insect repellents, respectively (Cetin et al. 2004; Lucantoni et al. 2006). Previously, different parts of medicinal plants such as leaves, seeds, peel, and succulent branches have shown great mosquitocidal properties (Ashfaq and Ashfaq 2012; Shaalan et al. 2005). In the current study, we have observed the female oviposition behavior and development period from hatching till adult emergence. This is the first study where trees extracts have been tested while previously mostly small plant extracts have been tested on mosquitoes. Moreover, mosquitoes might be more familiar with small vegetation like grasses and shrubs to hide during the day, while large trees like Eucalyptu, Neem and Moringa are something new and may induce behaviroal and physiological changes more effectively than other plants. For this purpose, we have used leaves and bark extracts of three tree species; Moringa oleifera, Azadirachta indica and Eucalyptus camaldulensis to explore their potential negative effects on the oviposition behavior and fitness as well as toxicity on immature stages of Cx. quinquefasciatus . Methods Insects Pupae as well as larvae of Cx. quinquefasciatus were collected from the sewage line of student hostel area of Bahauddin Zakariya University, Multan (30°5′11N, 71°39′15E), Punjab, Pakistan. The pupae and larvae were kept in plastic containers (2 L), containing the same sewerage water (1L) and covered at open end with a muslin cloth in laboratory till emergence of adults. After emergence, male and female adults were kept together in the Plexiglas cages (1×1×1 ft) for mating purpose and provided with 10% sugar solution for feeding provided on an ad libitum hydrated cotton pad. After 3-4 days, only females were given a pigeon with exposed keel for a period of 12 h (overnight) for blood feeding in dark conditions. After blood-feeding, blood-fed females were kept for next 48 h post-blood feeding prior to use in further experiments. All the experiments were conducted under standard laboratory conditions of temperature at 27±2 ºC, light: dark (12: 12h) photoperiod, and relative humidity 70±5% (Shah et al. 2017). Preparation of plant water extracts Water based plant extracts were prepared by following the methods used by Wakil et al. (2014) and Arunpandiyan (2011) with some modifications. Comprehensively, 500 g of fresh leaves as well as stem-bark of three different plant species ( Moringa oleifera, Azadirachta indica and Eucalyptus camaldulensis ) were collected separately in polythene bags. After collection, bark and leaves were grinded separately in mortar and pestle as well as in electric grinder (Black & Decker Company, New Britain, Connecticut, United States) by adding 500-800 mL of distilled water to make a uniform paste. Distilled water was added in the paste to make an amount of 2 L containing 500 gm of bark or leaves. Liquid mixtures of each botanical were poured in separate sterilized steel containers (5 L). These mixtures were stirred with a dried wooden stick continuously at an interval of 10-15 min for next three hours. After 3 h, mixtures were further extracted by using fine muslin cloth. After filtering the liquid, the remaining bark or leaf matter was pressed as much as possible to take out water from it. Furthermore, a standard solution (1000 mL) of 10% (w/v) for each type of extract was prepared by taking the required amount of filtrate and adding distilled water to prepare the solution of desired molarity. All the solutions were kept and maintained at 4 °C in the refrigerator. Oviposition bioassay For oviposition bioassays, 10 mL of 10% extract was added into a plastic cup (120 mL) containing 90 mL distilled water to make 1% (w/v) solution (i.e. 1 gm leaf weight in 99 mL water). Plastic cups containing 100 mL of different extracts (1%) i.e. Neem leaf (Neem L), Neem bark (Neem B), Eucalyptus leaf (Eucalyptus L), Eucalyptus bark (Eucalyptus B), Moringa leaf (Moringa L), and Moringa bark (Moringa B) were placed singly in adult cages as oviposition substrates and 10 blood-fed females were released in each cage including control containing only distilled water and 5 replications were completed for each and every extract type and control. Oviposition was observed on daily basis till death of last female in each treatment. Ovipositional substrates (botanical solution or distilled water in case of control) were replaced with the fresh ones at an interval of 48 h to avoid any fungal development. Number of egg rafts and number of eggs in each egg raft were counted (under a compound microscope having a magnification of 4X) on daily basis throughout the life span of females. Female longevity was also noted by observing the female mortality on daily basis till the last female died. Larval development Effects of tested plant extracts on the larval development of Cx. quinquefasciatus were also examined. An egg raft was placed in plastic cup containing 100 mL of 1% plant extract or the only distilled water (control). Larval emergence, larval mortality, pupae formation, and adult emergence were observed on daily basis (24 h) till all the larvae became pupae and adults. An egg raft was considered as a replicate and each treatment was replicated five times. Rearing mediums (extract or distilled water) were refreshed after every 48 h. Larvicide bioassays The larval toxicity of tested botanicals (mentioned above) was evaluated against the 24h, 48h and 72h old larvae of Cx. quinquefasciatus following the methodology described by World Health Organization (1996) with some modifications. Four concentrations of each botanical were prepared ranging from 100 ppm to 100,000 ppm. Each concentration was replicated five times. Ten (for each age group) larvae were used in each replicate, i.e., 50 larvae per concentration and 250 larvae in one bioassay including control. The control was consisted of only water. The data was recorded at 24h, 48h, 72h and 96h intervals. All those larvae which were unable to move were considered dead. Data analysis The Shapiro–Wilk test was applied to test the normality of the data and P -values of less than 0.05 were considered as significant (data set was taken as non-normal). Therefore, non-parametric test, Mann–Whitney U-test was used for pairwise comparison of different parameters of each treatment including delay in 1 st oviposition, female fecundity, larval emergence, larval development, pupation, adult emergence, larval survival, adult survival and longevity compared to that of control. For significance level, α was set at 5% and P -values of less than 0.05 were considered as significant. All the data were analyzed by using IBM SPSS Statistics v 21. The concentration response data was analyzed by probit analysis (Finney 1971) to determine the median lethal time (LT 50 ) values, their 95% confidence intervals (CI), slope ± standard error (SE) and chi square (χ 2 ). LT 50 values were considered significantly different when their 95% CI did not overlap (Litchfield and Wilcoxon 1949; Robertson and Preisler 1992). Pearson correlation was applied to determine the coefficient of correlation and significance of correlation ( P <0.05) between the LT 50 values and different age groups of exposed larvae at highest tested concentration of different botanicals. Results Delay in onset of 1 st oviposition The 1 st oviposition started earlier in control than all other treatments. The highly significant delay in oviposition was observed in Eucalyptus leaf (U=9, P =0.046) as compared to the control (Figure 1). Similarly, the 1 st oviposition was delayed with significant effect in Eucalyptus bark (U=9, P =0. 046), Neem leaf (U=9, P =0. 043), Neem bark (U=9, P =0.046), Moringa leaf (U=9, P =0.043) and Moringa bark (U=9, P =0.046) as compared to control (Figure 1). Female fecundity Number of egg rafts Numbers of egg rafts laid per female in Neem leaf (U=1, P =0.11), Moringa leaf (U=1.5, P =0.18) and Moringa bark (U=1, P =0.08) did not differ significantly than that of control (Figure 2). Females exposed to botanical extracts of Eucalyptus bark (U=0.0, P =0.05), Eucalyptus leaf (U=0, P =0.05) and Neem bark (U=0, P =0.05) laid similar number of egg rafts significantly lower as compared with that of control. Number of eggs/female Number of eggs laid by a female were significantly lower in Eucalyptus leaf (U=0.0, P =0.05), Eucalyptus bark (U=0.0, P =0.05), Neem leaf (U=0.0, P =0.05), Neem bark (U=0, P =0.05), Moringa leaf (U=0.0, P =0.05) and Moringa bark (U=0.0, P =0.05) compared to control (Figure 3). Larval emergence The larval emergence was maximum in the control and was significantly reduced in Neem leaf (U=0.0, P =0.05), Eucalyptus bark (U=0, P =0.05), Eucalyptus leaf (U=0, P =0.05) and in Moringa leaf (U=0, P =0.05). However, Neem bark (U=0, P =0.05) and Moringa bark (U=5, P =0.83) did not show any negative effect on larval emergence as compared to control (Figure 4). Pupation Highest pupation (%) in control was observed while no pupae were formed in Eucalyptus leaf (U=0.0, P =0.04) and Eucalyptus bark (U=0.0, P =0.04). Pupation (%) was significantly lower in Neem leaf (U=0.0, P =0.04) and Neem bark (U=0.0, P =0.05) as compared with control. A relatively higher proportion of pupae were formed in Moringa leaf (U=0.0, P =0.05) and Moringa bark (U=0.0, P =0.05) but significantly lower as compared to control (Figure 5) . Adult emergence As no pupae were formed in Eucalyptus leaves and bark, there was no adult emergence in Eucalyptus leaf (U=0.0, P =0.05) and bark (U=0.0, P =0.04). Moreover, no adults were emerged in Neem bark (U=0.0, P =0.04) (Figure 6). Adult emergence was significantly lower in Neem leaf extract (U=0.0, P =0.046) and Moringa leaf (U=0.0, P =0.05) as compared to the control. However, Moringa bark (U=2.0, P =0.28) did not reduce adult emergence of Cx. quinquefasciatus as compared with control (Figure 6). Lethal time (LT 50 ) of botanical extracts to different ages of larvae In case of 24h old larvae, the LT 50 values of the Neem leaf (100,000 PPM), Neem Bark (100,000 PPM), Moringa leaf (100,000 PPM), Eucalyptus leaf (100,000 PPM) were similar to each other based on the overlapping 95%CI and were the least time taking among all tested concentrations to kill the 50% population of the exposed larvae (Table 1). While, Neem leaf (100 PPM), Moringa leaf (100 PPM) and Eucalyptus leaf (100 PPM) were longest time taking concentrations among all the tested concentrations against 24h old larvae to kill the 50% population of the exposed larvae. Neem bark (100 PPM) and Moringa bark (100 PPM) did not kill any of the exposed population. When 48h larvae were exposed to different botanical concentrations, Neem leaf (100,000 PPM), Neem bark (100,000 PPM), Eucalyptus Leaf (100,000 PPM) and Eucalyptus bark were the concentrations having similar LT 50 based on overlapping 95% CI and the shortest time taking (Table 2). Moreover, the neem leaf (100 PPM) was the longest time taking concentration among all tested concentrations to kill fifty percent of the population. However, Moringa bark (100 PPM) did not kill any larvae exposed to it. In case of 72h old larvae, Neem leaf (100,000 PPM), Neem bark (100,000 PPM), Moringa Bark (100,000 PPM), Eucalyptus Leaf (100,000 PPM) and Eucalyptus bark (100,000 PPM) were similar based on overlapping 95% CI and lowest time taking concentrations among all the tested concentrations (Table 3). No mortality was observed at 100 PPM of all the tested botanicals and 1000 PPM of the Neem leaf, Moringa leaf, Moringa bark and Eucalyptus Bark. At highest tested concentration of every botanical, LT 50 was highly age dependent that is higher for elder larvae and vice versa (Table 4). Our results showed a strong positive but non-significant correlation between the different age groups and LT 50 values of the highest tested concentrations of the tested botanicals. Discussion Application of synthetic pesticides for the control of insect vectors is questioned due to resistance, environmental pollution and public health problems (Rose 2001). Whereas, various studies suggested botanical extracts are an effective tool for the control of insect pests (Mohan et al. 2006; Rahuman et al. 2008; Shahi et al. 2010). Demand for exploration of the eco-friendly control tactics against mosquitos is on rise day by day (Benelli et al. 2016). Some phytochemicals interfere with growth and development either with reproduction or acting as olfactory stimulus in form of repellent or attractant, while some act as common toxicants against larval and adult stages of mosquitoes (Shaalan et al. 2005). Our results suggest that onset of 1 st oviposition was delayed as compared with control, when females were exposed to each of tested botanical extracts. Previously, Rajkumar and Jebanesan (2009) have reported negative impact of Cassia obtusifolia on oviposition behavior of the mosquitoes. Mehra and Hiradhar (2002) reported that Cuscuta hyaline crude extract showed good oviposition deterrence against Cx. quinquefasciatus females. Moreover, Coria et al. (2008) also reported that Melia azidarach leaf extracts has deterrent effects on oviposition of Ae. aegypti . Our results are also in line with that of Govindarajan et al. (2011b) who has shown that the leaf extracts of Caesalpinia pulcherrima and Ervatamia coronariaand have activities (larvicidal and ovicidal) against Anopheles stephensi, Ae. aegypti, and Cx. quinquefasciatus . Similar findings has been reported by Elango et al. (2009) that indigenous plant extracts such as Aegle marmelos, Andrographis lineate, Andrographis paniculata, Cocculus hirsutus, Eclipta prostrata , and Tagetes erecta have oviposition-deterrent, ovicidal, and repellent activity against An. subpictus . As oviposition was significantly delayed in exposed females in all treatments, oviposition deterrent activity of plant extracts might induce physiological changes leading to behavioral avoidance in female adult mosquitoes. It might also be due to volatile chemicals released from respective plant water extracts, which were added in their oviposition substrates. Tawatsin et al. (2006) has also reported that the relatively high oviposition deterrent activity was induced by essential oils of Curcuma longa, Zingiber officinale , Vitex trifolia, Melaleuca cajuputi , Manglietia garrettii , and Houttuynia cordata . The delay in oviposition leads to lower female fecundity and thus it is obvious that with vast biodiversity, many bioactive compounds could be isolated and utilized for mosquito control especially Cx. quinquefasciatus . Moreover, adding simple water extracts of potential botanicals in the standing waters in ditches, ponds or in sewerage lines, which are the main breeding sites of Cx. quinquefasciatus , may help to deter gravid females or delaying the oviposition leading to less female fecundity and ultimately less population of Cx. quinquefasciatus . Larval emergence was significantly lower in Neem leaf, Eucalyptus leaf, Eucalyptus bark and Neem bark. Prathibha et al. (2014) found that the ethyl acetate extracts of S. mauritiana have marked effect results in zero hatchability at 100 ppm against An. stephensi , Ae. aegypti , and Cx. quinquefasciatus. Previously, M. oleifera seed water extract has been reported to exhibit 80%, 41.3% and 9.33% larval mortality at 5%, 2.5% and 1% concentrations on larvae of Anopheles gambiae (NJOM et al. 2011) . At higher concentration (5% and 2.5%), no larvae reached to pupal stage but at lower concentration even few adults were emerged. Similarly,Tandon and Sirohi (2010) reported that A. indica aqueous crude leaf extract caused 70-99% larval mortality followed by Gymnema sylvestre 44-89%, Nerium indicum 41-74% and Datura metel 19-54% mortality of larvae. An inverse impact of botanical extracts on hatchability of eggs was observed against Cx. quinquefasciatus (Govindarajan et al. 2011a). Hence, it can be concluded that unprocessed water extracts of tested plant have more potential to suppress the life table of target insects due to presence of more diverse compounds (Naumann and Isman 1995). In the present study, no pupation occurred in Eucalyptus leaf and in Eucalyptus bark, while very few pupae were formed in Neem leaf and Neem bark. These results are similar to NJOM et al. (2011) where higher concentration of Moringa seed extracts did not allow any pupation. Previously, hexane based leaves extract of Eucalyptus has shown potential to suppress the pupal formation in from larvae of different exposed mosquito species including Cx. quinquefasciatus (Singh et al. 2007). This also indicates that plant extracts can kill the larvae as well as pupae of mosquitoes. In the present study, no adult emergence in Eucalyptus leaf, Eucalyptus bark and Neem bark while was highly significantly lower in Neem leaf. Elimam et al. (2009) reported that aqueous extracts from Ricinus communis inhibit adult emergence and also showed larvicidal as well as oviposition deterrent effect against Cx. quinquefasciatus and An. arabiensis. Conclusions The water based botanical extracts of bark and leaves of the all tested species showed a variable range of larvicidal activity against the larvae of the Cx. quinquefasciatus but have potential to be used as the traditional control agents . Among the tested botanicals, mortality of the larvae was highly dose dependent. Highest mortality at higher doses and lowest or no mortality was observed at lowest doses of the tested botanicals in our study. Previously, Govindarajan et al. (2011b) reported the similar dose dependent response of the Cx. quinquefasciatus to botanicals. Cetin et al. (2006) reported a similar dose dependent response of Cx. pipens exposed to plant extracts. Therefore, application, keeping in view the dose response of the target species, of these easily made botanical extracts shall be considered at least at small scale level, e.g., homes and poultry farms. Declarations Ethics approval and consent to participate: No human beings were subject of experiments; therefore, no consent to participate was required. Consent for publication: No human beings were subject of experiments; therefore, no consent to Publish was required Availability of data and material: Data will be available on demand Author Contribution: MK and RMS performed the experiment, SA, SM, SA and MB designed the experiment, provided materials and guidelines to perform the experiments. MK and RMS wrote the MS. SA, SM, SA and MB critically reviewed and finalized it. Funding: The authors did not receive support from any organization for the submitted work Competing interests: The authors hereby declared no competing interests Acknowledgements: All the authors are cordially thankful to laboratory staff in helping to perform labor work during experimentation. References Arunpandiyan G (2011) Toxicity of aqueous crude neem leaf extract against Culex Int J Pharm Biomed Sci 2(1):1-3 Ashfaq M, Ashfaq U (2012) Evaluation of mosquitocidal activity of water extract of Moringa oleifera seeds against Culex quinquefasciatus (Diptera: Culicidae) in Pakistan. Pak Entomol 34(1):21-26 Benelli G, Jeffries CL, Walker T (2016) Biological control of mosquito vectors: past, present, and future. Insects 7(4):52 Borah R, Kalita M, Kar A, Talukdar A (2010) Larvicidal efficacy of Toddalia asiatica (Linn.) Lam against two mosquito vectors Aedes aegypti and Culex quinquefasciatus . African Journal of Biotechnology 9(17):2527-2530 Cetin H, Cinbilgel I, Yanikoglu A, Gokceoglu M (2006) Larvicidal activity of some Labiatae (Lamiaceae) plant extracts from Turkey. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives 20(12):1088-1090 Cetin H, Erler F, Yanikoglu A (2004) Larvicidal activity of a botanical natural product, AkseBio2, against Culex pipiens . Fitoterapia 75(7):724-728 Coria C, et al. (2008) Larvicide and oviposition deterrent effects of fruit and leaf extracts from Melia azedarach on Aedes aegypti (L.)(Diptera: Culicidae). Bioresource Technology 99(8):3066-3070 Elango G, Rahuman AA, Bagavan A, Kamaraj C, Zahir AA, Venkatesan C (2009) Laboratory study on larvicidal activity of indigenous plant extracts against Anopheles subpictus and Culex tritaeniorhynchus . Parasitol Res 104(6):1381-1388 Elimam AM, Elmalik KH, Ali FS (2009) Larvicidal, adult emergence inhibition and oviposition deterrent effects of foliage extract from Ricinus communis against Anopheles arabiensis and Culex quinquefasciatus in Sudan. Trop Biomed 26(2):130-139 Finney D (1971) A statistical treatment of the sigmoid response curve. Probit analysis, 3rd edn Cambridge University Press, London 333 Govindarajan M, Mathivanan T, Elumalai K, Krishnappa K, Anandan A (2011a) Mosquito larvicidal, ovicidal, and repellent properties of botanical extracts against Anopheles stephensi , Aedes aegypti , and Culex quinquefasciatus (Diptera: Culicidae). Parasitology Research 109(2):353-367 Govindarajan M, Mathivanan T, Elumalai K, Krishnappa K, Anandan A (2011b) Ovicidal and repellent activities of botanical extracts against Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi (Diptera: Culicidae). Asian Pacific Journal of Tropical Biomedicine 1(1):43-48 Guedes DR, et al. (2017) Zika virus replication in the mosquito Culex quinquefasciatus in Brazil. Emerging Microbes & Infections 6(1):1-11 Guessan R, Corbel V, Akogbéto M, Rowland M (2007) Reduced efficacy of insecticide-treated nets and indoor residual spraying for malaria control in pyrethroid resistance area, Benin. Emerging Infect Dis 13(2):199 Hafeez F, Akram W, Shaalan EA-S (2011) Mosquito larvicidal activity of citrus limonoids against Aedes albopictus . Parasitol Res 109(1):221-229 Kilpatrick AM, Kramer LD, Jones MJ, Marra PP, Daszak P (2006) West Nile virus epidemics in North America are driven by shifts in mosquito feeding behavior. PLoS Biol 4(4):e82 Litchfield JT, Wilcoxon F (1949) A simplified method of evaluating dose-effect experiments. J Pharmacol Exp Ther 96(2):99-113 Lucantoni L, et al. (2006) Effects of a neem extract on blood feeding, oviposition and oocyte ultrastructure in Anopheles stephensi Liston (Diptera: Culicidae). Tissue and Cell 38(6):361-371 Mehra B, Hiradhar P (2002) Cuscuta hyalina Roth., an insect development inhibitor against common house mosquito Culex quinquefasciatus J Environ Biol 23(3):335-339 Mohan L, Sharma P, Srivastava C (2006) Evaluation of Solanum xanthocarpum extract as a synergist for cypermethrin against larvae of the filarial vector Culex quinquefasciatus (Say). Entomological Research 36(4):220-225 Naumann K, Isman MB (1995) Evaluation of neem Azadirachta indica seed extracts and oils as oviposition deterrents to noctuid moths. Entomologia experimentalis et applicata 76(2):115-120 NJOM VS, UMEH EDN, EZE CS (2011) MALARIA VECTOR Anopheles gambiae . Animal Research International 8(2):1392-1397 Okumu FO, Moore SJ, Okumu F, Moore S (2011) Combining indoor residual spraying and insecticide-treated nets for malaria control in Africa: a review of possible outcomes and an outline of suggestions for the future. Malaria journal 10(208):1-13 Prathibha K, Raghavendra B, Vijayan V (2014) Larvicidal, ovicidal, and oviposition-deterrent activities of four plant extracts against three mosquito species. Environmental Science and Pollution Research 21(10):6736-6743 Rahuman AA, Gopalakrishnan G, Venkatesan P, Geetha K (2008) Larvicidal activity of some Euphorbiaceae plant extracts against Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae). Parasitol Res 102(5):867-873 Rajkumar S, Jebanesan A (2009) Larvicidal and oviposition activity of Cassia obtusifolia Linn (Family: Leguminosae) leaf extract against malarial vector, Anopheles stephensi Liston (Diptera: Culicidae). Parasitol Res 104(2):337-340 Robertson J, Preisler H (1992) Pesticide Bioassays with Arthropods. CRC, Boca Raton, FL Rose RI (2001) Pesticides and public health: integrated methods of mosquito management. Emerging Infect Dis 7(1):17 Sangshetti JN, Shinde DB, Kulkarni A, Arote R (2017) Two decades of antifilarial drug discovery: a review. RSC advances 7(33):20628-20666 Shaalan EA-S, Canyon D, Younes MWF, Abdel-Wahab H, Mansour A-H (2005) A review of botanical phytochemicals with mosquitocidal potential. Environ Int 31(8):1149-1166 Shah RM, et al. (2017) Larval Habitat Substrates Could Affect the Biology and Vectorial Capacity of Culex quinquefasciatus (Diptera: Culicidae). J Med Entomol 54(3):638-645 Shahi M, Hanafi-Bojd A, Iranshahi M, Vatandoost H, Hanafi-Bojd M (2010) Larvicidal efficacy of latex and extract of Calotropis procera (Gentianales: Asclepiadaceae) against Culex quinquefasciatus and Anopheles stephensi (Diptera: Culicidae). J Vector Borne Dis 47(3):185-188 Simonsen PE, Mwakitalu ME (2013) Urban lymphatic filariasis. Parasitol Res 112(1):35-44 Singh R, Dhiman R, Mittal P (2007) Studies on mosquito larvicidal properties of Eucalyptus citriodora Hook (family-Myrtaceae). The Journal of Communicable Diseases 39(4):233-236 Tahir HM, Butt A, Khan SY (2009) Response of Culex quinquefasciatus to deltamethrin in Lahore district. J Parasitol Vector Biol 1(3):19-24 Tandon P, Sirohi A (2010) Assessment of larvicidal properties of aqueous extracts of four plants against Culex quinquefasciatus Jordan Journal of Biological Sciences 147(612):1-10 Tawatsin A, et al. (2006) Repellency of essential oils extracted from plants in Thailand against four mosquito vectors (Diptera: Culicidae) and oviposition deterrent effects against Aedes aegypti (Diptera: Culicidae). Southeast Asian J Trop Med Public Health 37(5):915-31 Van Bortel W, et al. (2008) The insecticide resistance status of malaria vectors in the Mekong region. Malaria journal 7(1):102 Wakil W, Ghazanfar M, Kwon Y, Ullah E, Islam S, Ali K (2014) Testing Paecilomyces lilacinus, diatomaceous earth and Azadirachta indica alone and in combination against cotton aphid (Aphis gossypii Glover)(Insecta: Homoptera: Aphididae). African Journal of Biotechnology 11(4):821-828 World Health Organization W (1996) Report of the WHO informal consultation on the evaluation and testing of insecticides, CTD/WHO PES/IC/ 1996; 96.1: 69. Tables Table 1 . Lethal time (LT 50 ) values of the 24h old larvae of Culex quinquefasciatus exposed to different botanicals Botanical Extract Concentration (PPM) LT a1 50 (95%CI b ) Slope ( ±SE) x2 c Df d P Neem leaf 100 184.312 (137.69-481.184) 0.017±0.006 1.475 2 0.478 1000 107.164 (81.902-1636.850) 0.022±0.003 4.529 2 0.104 10000 69.976 (44.343-120.657) 0.025±0.003 5.902 2 0.052 100000 8.257 (17.252-21.968) 0.016±0.003 0.089 2 0.956 Neem Bark 100 A A A A A 1000 95.819 (75.44-247.67) 0.025±0.003 4.658 2 0.097 10000 66.338 (61.365-71.625) 0.027±0.003 1.714 2 0.424 100000 28.52 (20.468-34.481) 0.029±0.003 3.098 2 0.212 Moringa leaf 100 150.372 (120.38-715.81) 0.032±0.015 0.13 2 0.937 1000 106.39 (96.33-122.95) 0.022±0.003 1.867 2 0.393 10000 75.232(68.402-83.20) 0.020 ±0.003 0.729 2 0.694 100000 11.16 (8.938-22.844) 0.019 ±0.003 1.37 2 0.504 Moringa Bark 100 A A A A A 1000 112.95(a) 0.023±0.004 4.847 2 0.089 10000 92.88 (84.145-106.24) 0.019±0.003 3.791 2 0.15 100000 41.24(33.735-47.240) 0.024 ±0.003 2.743 2 0.254 Eucalyptus Leaf 100 140.39(119.35-198.78) 0.024 ± 0.006 1.289 2 0.525 1000 94.34 (86.39-106.13) 0.022 ± 0.003 3.604 2 0.165 10000 51.778 (46.67-56.50) 0.029±0.003 2.360 2 0.307 100000 15.528(4.487-22.442) 0.036 ± 0.005 0.528 2 0.768 Eucalyptus Bark 100 191.880 (a) 0.021 ± 0.014 0.435 2 0.805 1000 94.343 (86.388-106.128) 0.022 ±0.003 3.604 2 0.165 10000 57.889(53.095-62.572) 0.029± 0.003 2.02 2 0.364 100000 34.738 (29.421-39.136) 0.037±0.004 0.076 2 0.963 “A/a”indicates that software was unable to calculate the value, “A/a”indicates that software was unable to calculate the value, a1=lethal time in hours, CI=confidence intervals, X 2 =chi-square goodness of fit test, df=degree of freedom Table 2. Lethal time (LT 50 ) values of the 48h old larvae of Culex quinquefasciatusa exposed to different botanicals Botanical Extract Concentration (PPM) LT a1 50 (95%CI b ) Slope ( ±SE) x2 c Df d P Neem leaf 100 545.688 (a) 0.005±0.006 0.141 2 0.932 1000 157.066 (124.926-360.985) 0.024±0.009 1.234 2 0.539 10000 71.049 (64.155-79.284) 0.019±0.003 0.759 2 0.684 100000 50.351 (44.858-55.319) 0.027±0.003 0.419 2 0.811 Neem Bark 100 191.888(a) 0.021±0.014 0.435 2 0.805 1000 169.160 (a) 0.025±0.014 0.268 2 0.874 10000 70.149 (64.385-76.722) 0.023±0.003 0.251 2 0.882 100000 54.006 (49.271-58.496) 0.031±0.003 0.725 2 0.696 Moringa leaf 100 123.14 (a) 0.085 ±0.372 0.001 2 0.999 1000 123.14 (a) 0.085±0.372 0.001 2 0.999 10000 88.49 (a) 0.022±0.003 8.512 2 0.014 100000 47.54 (a) 0.020± 0.003 9.935 2.000 0.007 Moringa Bark 100 A A A A a 1000 123.141 (a) 0.085±0.372 0.001 2 0.999 10000 115.256 ( 88.925-6217.97) 0.023 ±0.004 3.971 2 0.137 100000 72.400 (65.617-80.624) 0.019 ±0.003 2.426 2 0.297 Eucalyptus Leaf 100 285.803 (a) 0.011 ± 0.008 0.653 2 0.721 1000 207.524 (142.97-2516.20) 0.015 ± 0.007 0.405 2 0.817 10000 63.864 (57.784-70.262) 0.022 ± 0.003 1.383 2 0.501 100000 47.915 (12.16-68.37) 0.030 ± 0.003 6.1 2 0.047 Eucalyptus Bark 100 191.88 (a) 0.021 ±0.014 0.435 2 0.805 1000 339.974(a) 0.007±0.005 .086 2 0.903 10000 69.56 (63.327-76.703) 0.021 ± 0.003 1.963 2 0.375 100000 45.524 (40.361-50.093) 0.031± 0 .003 0.292 2 0.864 “A/a”indicates that software was unable to calculate the value, “A/a”indicates that software was unable to calculate the value, a1= a1=lethal time in hours, CI=confidence intervals, X 2 =chi-square goodness of fit test, df=degree of freedom Table 3.Lethal time (LT 50 ) values of the 72h old larvae of Culex quinquefasciatus exposed to different botanicals Botanical Extract Concentration (PPM) LT a1 50 (95%CI b ) Slope ( ±SE) x2 c Df d P Neem leaf 100 A a a A a 1000 A a a A a 10000 136.869 (116.511-182.905) 0.018±0.004 1.925 2 0.382 100000 61.404 (40.060-83.379) 0.028±0.003 4.761 2 0.093 Neem Bark 100 A a a A a 1000 114.102 (a) 0.114±0.913 <0.0001 2 1 10000 131.090 (a) 0.021±0.004 6.04 2 0.049 100000 62.932 (30.964-99.006) 0.029±0.003 7.476 2 0.024 Moringa leaf 100 A a a A a 1000 A a a A a 10000 131.09 (a) 0.021± 0.004 6.04 2 0.049 100000 56.106 (a) 0.023± 0.003 12.609 2 0.002 Moringa Bark 100 A a a A a 1000 A a a A a 10000 157.18 (a) 0.014 ± 0.004 7.024 2 0.03 100000 80.014(72.948-89.394) 0.020 ± 0.003 0.88 2 0.644 Eucalyptus Leaf 100 123.141 (a) 0.085 ±0.372 0.001 2 0.999 1000 169.79 (127.99-981.16) 0.023 ±0.011 0.843 2 0.656 10000 100.175 (81.453-210.426) 0.027 ±0.004 3.879 2 0.144 100000 59.590(54.563-64.613) 0.027 ±0 .003 1.427 2 0.49 Eucalyptus Bark 100 A a a A a 1000 123.141(a) 0.085±0 .372 0.001 2 0.999 10000 102.028 (92.181-117.970) 0.020± 0.003 1.825 2 0.401 100000 62.136 (56.978-67.410) 0.026± 0.003 0.155 2 0.925 “A/a”indicates that software was unable to calculate the value, a1=lethal time in hours, CI=confidence intervals, X 2 =chi-square goodness of fit test, df=degree of freedom Table 4. Correlation between lethal time (LT 50 ) values and different age groups of larvae of Culex quinquefasciatus exposed to highest tested concentration of different botanicals Botanical Extracts Concentration (PPM) Larval age (h) LT a1 50 (95%CI b ) Correlation Neem Leaf 100000 24 8.257 (17.252-21.968) (r c =0.95, P d =0.21) 48 50.351 (44.858-55.319) 72 61.404 (40.060-83.379) Neem Bark 100000 24 28.52 (20.468-34.481) (r=0.96, P =0.17) 48 54.006 (49.271-58.496) 72 62.932 (30.964-99.006) Moringa leaf 100000 24 11.16 (8.938-22.844) (r=0.94, P =0.22) 48 47.54 (a) 72 56.106 (a) Moringa Bark 100000 24 41.24(33.735-47.240) (r=0.94, P =0.21) 48 72.400 (65.617-80.624) 72 80.014(72.948-89.394) Eucalyptus Leaf 100000 24 15.528(4.487-22.442) (r=0.97, P =0.17) 48 47.915 (12.16-68.37) 72 59.590(54.563-64.613) Eucalyptus Bark 100000 24 34.738 (29.421-39.136) (r=0.99, P =0.08) 48 45.524 (40.361-50.093) 72 62.136 (56.978-67.410) “a” indicate that software was unable to calculate the value, a1=lethal time in hours, CI=confidence intervals, C=coefficient of correlation, P =Significance of correlation ( P >0.05) Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-143809","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research","associatedPublications":[],"authors":[{"id":7858642,"identity":"f7a80987-f5ed-4c21-b91a-2eccfb5329c4","order_by":0,"name":"Muhammad Khan","email":"","orcid":"","institution":"bzu","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"Khan","suffix":""},{"id":7858643,"identity":"5608fd85-2f54-4552-9ee6-3d941d3ca999","order_by":1,"name":"Rizwan shah","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABCUlEQVRIiWNgGAWjYDACZiT2B4YKCTkQ48ADorSwMTDOYDhjYwzWkkCUdSAtjG1piQ0gDj4t5u3szyR+7tiWOH9+88OGH2yH0+eHHX4ItMVOTrcBuxaZwwxpkr1nbiduOMZm2NjDczh34+00A6CWZGOzA9i1SDAzHLvB2wbUwsZg/oBHAqhldgJIy4HEbTi1MLbd/AvUMr+N/WPjH4PD6Yaz0z8Q0MLMdhtkS8MxHsNmnoS0BHnpHEK2sLH/lm27bbzhWE5hs8wBG8MN0jkFBxIM8PiF//hjw7dtt2XnNx/f2Pj2n4S8/Oz0zR8+VNjJ4dKCCQzAKg2IVQ4C8g2kqB4Fo2AUjIKRAADZSWaOKarqDgAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-7168-5700","institution":"Bahauddin Zakariya University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Rizwan","middleName":"","lastName":"shah","suffix":""},{"id":7858644,"identity":"8518964a-5007-449b-8d13-5b3b5fd1da58","order_by":2,"name":"Sarfraz Ali Shad","email":"","orcid":"","institution":"BZU: Bahauddin Zakariya University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Sarfraz","middleName":"Ali","lastName":"Shad","suffix":""},{"id":7858645,"identity":"25090910-f91e-4c4f-a1a7-1891e3430616","order_by":3,"name":"Shahid Majeed","email":"","orcid":"","institution":"BZU: Bahauddin Zakariya University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Shahid","middleName":"","lastName":"Majeed","suffix":""},{"id":7858646,"identity":"05bf0a78-2b4a-42ef-bd89-23118d2e13dc","order_by":4,"name":"Shahbaz Ahmad","email":"","orcid":"","institution":"BZU: Bahauddin Zakariya University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Shahbaz","middleName":"","lastName":"Ahmad","suffix":""},{"id":7858647,"identity":"ebdcff6d-1e36-41c2-bf26-a573fa0e5c78","order_by":5,"name":"Muhammad Binyameen","email":"","orcid":"","institution":"BZU: Bahauddin Zakariya University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"Binyameen","suffix":""}],"badges":[],"createdAt":"2021-01-08 23:46:54","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-143809/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-143809/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":5051436,"identity":"0b058fab-01ac-4fb2-b29c-771252e91fc4","added_by":"auto","created_at":"2021-01-18 15:59:47","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":12921,"visible":true,"origin":"","legend":"Effects of different plant water extracts in delaying 1st oviposition in Culex quinquefasciatus. *, ** and *** indicate the significant delay in those treatments compared to the control at P\u003c0.05, P \u003c0.005 and, P \u003c0.0001 respectively. “L” and “B” denote leaf and bark, respectively. The error bars represent the standard error of the mean.","description":"","filename":"Fig01.png","url":"https://assets-eu.researchsquare.com/files/rs-143809/v1/aa11231380577ab8d6895028.png"},{"id":5051532,"identity":"252469cc-82b0-43ad-9fee-950b79d59b95","added_by":"auto","created_at":"2021-01-18 16:02:47","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":49495,"visible":true,"origin":"","legend":"Effect of different botanicals on the production of egg rafts in Culex quinquefasciatus. *, ** and *** indicate the significant delay in those treatments compared to the control at P\u003c0.05, P \u003c0.005 and, P \u003c0.0001, respectively. The error bars represent the standard error of the mean. ","description":"","filename":"Fig02.png","url":"https://assets-eu.researchsquare.com/files/rs-143809/v1/8874e645ca581a50707b44f8.png"},{"id":5051669,"identity":"e90befaf-f8d6-412d-854c-20a44d7ec5e8","added_by":"auto","created_at":"2021-01-18 16:05:47","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":12349,"visible":true,"origin":"","legend":"Effects of different plant water extracts on the fecundity of the Culex quinquefasciatus. *, ** and *** indicate the significant delay in those treatments compared to the control at P\u003c0.05, P \u003c0.005 and, P \u003c0.0001 respectively. The error bars represent the standard error of the mean. ","description":"","filename":"Fig03.png","url":"https://assets-eu.researchsquare.com/files/rs-143809/v1/68525e61e150f44c475d5842.png"},{"id":5051531,"identity":"5ed4f25c-aae9-41d7-b797-0f3af15adc8b","added_by":"auto","created_at":"2021-01-18 16:02:47","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":18793,"visible":true,"origin":"","legend":"Effects of different plant water extracts on the larval emergence of the Culex quinquefasciatus. *, ** and *** indicate the significant delay in those treatments compared to the control at P\u003c0.05, P \u003c0.005 and, P \u003c0.0001, respectively. The error bars represent the standard error of the mean.","description":"","filename":"Fig04.png","url":"https://assets-eu.researchsquare.com/files/rs-143809/v1/f6a30057a303540382c2797c.png"},{"id":5051437,"identity":"456aaa89-b95d-45a1-8b2a-873c0e93fbc6","added_by":"auto","created_at":"2021-01-18 15:59:47","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":8796,"visible":true,"origin":"","legend":"Effects of different plant water extracts on the pupation (%) of the Culex quinquefasciatus. *, ** and *** indicate the significant delay in those treatments compared to the control at P\u003c0.05, P \u003c0.005 and, P \u003c0.0001 respectively. The error bars represent the standard error of the mean. ","description":"","filename":"Fig05.png","url":"https://assets-eu.researchsquare.com/files/rs-143809/v1/38da4c41bd4c32e4406ff32d.png"},{"id":5051534,"identity":"b3110206-2bb7-4cec-94aa-0ed831dec44a","added_by":"auto","created_at":"2021-01-18 16:02:47","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":11845,"visible":true,"origin":"","legend":"Effects of different plant water extracts on adult emergence of the Culex quinquefasciatus. *, ** and *** indicate the significant delay in those treatments compared to the control at P\u003c0.05, P \u003c0.005 and, P \u003c0.0001 respectively. The error bars represent the standard error of the mean. ","description":"","filename":"Fig06.png","url":"https://assets-eu.researchsquare.com/files/rs-143809/v1/aeefb8475f438bc1614b56f5.png"},{"id":13647220,"identity":"89cc5a0c-5570-4b3d-92a0-af755e210811","added_by":"auto","created_at":"2021-09-17 09:27:15","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":667602,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-143809/v1/d537841d-d2b4-47c1-9381-95cae7cfa336.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eWater Extracts From Trees Negatively Affect Oviposition Behavior and Fitness of Southern House Mosquito, Culex Quinquefasciatus Say (Diptera: Culicidae): A Simple, Easily Adoptable and Cost-Effective Approach to Control Mosquitoes\u003c/p\u003e","fulltext":[{"header":"Background","content":"\u003cp\u003eSouthern house mosquito, \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e Say, is a vector of many zoonotic diseases that affect humans and wild and domestic animals, such as avian malaria, Western equine encephalitis, lymphatic filariasis, St. Louis encephalitis, West Nile fever and Zika virus (Guedes et al. 2017; Simonsen and Mwakitalu 2013). Mortality rate due to vector-borne filariasis is low but a high morbidity rate is a social stigma. According to estimates, about 120 million people in 81 countries are currently infected, and 1.34 billion people who live in areas endemic to filariasis are at risk of infection (Sangshetti et al. 2017). In Pakistan, population of \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e develops rapidly during rainy season and becomes a sever biting nuisance leading to restless nights (Tahir et al. 2009). Although, the ornithophilic behavior of \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e is well reported, but the preference changes to mammals in the absence of primary host, which leads to significant impact (Kilpatrick et al. 2006).\u003c/p\u003e\n\u003cp\u003eApplication of synthetic insecticides as larvicides, insecticide-treated nets and indoor residual spraying (IRS) are the principal mosquito management measures adapted worldwide (Okumu et al. 2011). So far, chemical insecticides are the most effective control measures against mosquitoes, however, resistance against these chemical insecticides has recently been reported in many species of mosquitoes, which is resulting in repeated epidemic (Guessan et al. 2007; Van Bortel et al. 2008). Moreover, residual effects of synthetic insecticides are imposing serious threat to humans, animals, plants health and environment (References?). These major concerns have diverted the attention of researchers towards natural chemicals, such as plant extracts or oils, which are cost effective, safe, target specific and easily biodegradable in the environment (Borah et al. 2010; Hafeez et al. 2011).\u003c/p\u003e\n\u003cp\u003eIt has been shown by several studies that plant extracts or their non-volatile and volatile constituents have a great potential to be used as insecticides and insect repellents, respectively (Cetin et al. 2004; Lucantoni et al. 2006). Previously, different parts of medicinal plants such as leaves, seeds, peel, and succulent branches have shown great mosquitocidal properties (Ashfaq and Ashfaq 2012; Shaalan et al. 2005). In the current study, we have observed the female oviposition behavior and development period from hatching till adult emergence. This is the first study where trees extracts have been tested while previously mostly small plant extracts have been tested on mosquitoes. Moreover, mosquitoes might be more familiar with small vegetation like grasses and shrubs to hide during the day, while large trees like Eucalyptu, Neem and Moringa are something new and may induce behaviroal and physiological changes more effectively than other plants. For this purpose, we have used leaves and bark extracts of three tree species; \u003cem\u003eMoringa oleifera, Azadirachta indica\u003c/em\u003e and \u003cem\u003eEucalyptus camaldulensis \u003c/em\u003eto explore their potential negative effects on the oviposition behavior and fitness as well as toxicity on immature stages of \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e.\u003c/p\u003e"},{"header":"Methods","content":"\u003ch2\u003eInsects\u003c/h2\u003e\n\u003cp\u003ePupae as well as larvae of \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e were collected from the sewage line of student hostel area of Bahauddin Zakariya University, Multan (30\u0026deg;5\u0026prime;11N, 71\u0026deg;39\u0026prime;15E), Punjab, Pakistan. The pupae and larvae were kept in plastic containers (2 L), containing the same sewerage water (1L) and covered at open end with a muslin cloth in laboratory till emergence of adults. After emergence, male and female adults were kept together in the Plexiglas cages (1\u0026times;1\u0026times;1 ft) for mating purpose and provided with 10% sugar solution for feeding provided on an \u003cem\u003ead libitum\u003c/em\u003e hydrated cotton pad. After 3-4 days, only females were given a pigeon with exposed keel for a period of 12 h (overnight) for blood feeding in dark conditions. After blood-feeding, blood-fed females were kept for next 48 h post-blood feeding prior to use in further experiments. All the experiments were conducted under standard laboratory conditions of temperature at 27\u0026plusmn;2 \u0026ordm;C, light: dark (12: 12h) photoperiod, and relative humidity 70\u0026plusmn;5% (Shah et al. 2017).\u003c/p\u003e\n\u003ch2\u003ePreparation of plant water extracts\u003c/h2\u003e\n\u003cp\u003eWater based plant extracts were prepared by following the methods used by Wakil et al. (2014) and Arunpandiyan (2011) with some modifications. Comprehensively, 500 g of fresh leaves as well as stem-bark of three different plant species (\u003cem\u003eMoringa oleifera, Azadirachta indica \u003c/em\u003eand\u003cem\u003e Eucalyptus camaldulensis\u003c/em\u003e) were collected separately in polythene bags. After collection, bark and leaves were grinded separately in mortar and pestle as well as in electric grinder (Black \u0026amp; Decker Company, New Britain, Connecticut, United States) by adding 500-800 mL of distilled water to make a uniform paste. Distilled water was added in the paste to make an amount of 2 L containing 500 gm of bark or leaves. Liquid mixtures of each botanical were poured in separate sterilized steel containers (5 L). These mixtures were stirred with a dried wooden stick continuously at an interval of 10-15 min for next three hours. After 3 h, mixtures were further extracted by using fine muslin cloth. After filtering the liquid, the remaining bark or leaf matter was pressed as much as possible to take out water from it. Furthermore, a standard solution (1000 mL) of 10% (w/v) for each type of extract was prepared by taking the required amount of filtrate and adding distilled water to prepare the solution of desired molarity. All the solutions were kept and maintained at 4 \u0026deg;C in the refrigerator.\u003c/p\u003e\n\u003ch2\u003eOviposition bioassay\u003c/h2\u003e\n\u003cp\u003eFor oviposition bioassays, 10 mL of 10% extract was added into a plastic cup (120 mL) containing 90 mL distilled water to make 1% (w/v) solution (i.e. 1 gm leaf weight in 99 mL water). Plastic cups containing 100 mL of different extracts (1%) i.e. Neem leaf (Neem L), Neem bark (Neem B), Eucalyptus leaf (Eucalyptus L), Eucalyptus bark (Eucalyptus B), Moringa leaf (Moringa L), and Moringa bark (Moringa B) were placed singly in adult cages as oviposition substrates and 10 blood-fed females were released in each cage including control containing only distilled water and 5 replications were completed for each and every extract type and control. Oviposition was observed on daily basis till death of last female in each treatment. Ovipositional substrates (botanical solution or distilled water in case of control) were replaced with the fresh ones at an interval of 48 h to avoid any fungal development. Number of egg rafts and number of eggs in each egg raft were counted (under a compound microscope having a magnification of 4X) on daily basis throughout the life span of females. Female longevity was also noted by observing the female mortality on daily basis till the last female died.\u003c/p\u003e\n\u003ch2\u003eLarval development\u003c/h2\u003e\n\u003cp\u003eEffects of tested plant extracts on the larval development of \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e were also examined. An egg raft was placed in plastic cup containing 100 mL of 1% plant extract or the only distilled water (control). Larval emergence, larval mortality, pupae formation, and adult emergence were observed on daily basis (24 h) till all the larvae became pupae and adults. An egg raft was considered as a replicate and each treatment was replicated five times. Rearing mediums (extract or distilled water) were refreshed after every 48 h.\u003c/p\u003e\n\u003ch2\u003eLarvicide bioassays\u003c/h2\u003e\n\u003cp\u003eThe larval toxicity of tested botanicals (mentioned above) was evaluated against the 24h, 48h and 72h old larvae of \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e following the methodology described by World Health Organization (1996) with some modifications. Four concentrations of each botanical were prepared ranging from 100 ppm to 100,000 ppm. Each concentration was replicated five times. Ten (for each age group) larvae were used in each replicate, i.e., 50 larvae per concentration and 250 larvae in one bioassay including control. The control was consisted of only water. The data was recorded at 24h, 48h, 72h and 96h intervals. All those larvae which were unable to move were considered dead.\u003c/p\u003e\n\u003ch2\u003eData analysis\u003c/h2\u003e\n\u003cp\u003eThe Shapiro\u0026ndash;Wilk test was applied to test the normality of the data and \u003cem\u003eP\u003c/em\u003e-values of less than 0.05 were considered as significant (data set was taken as non-normal). Therefore, non-parametric test, Mann\u0026ndash;Whitney U-test was used for pairwise comparison of different parameters of each treatment including delay in 1\u003csup\u003est\u003c/sup\u003e oviposition, female fecundity, larval emergence, larval development, pupation, adult emergence, larval survival, adult survival and longevity compared to that of control. For significance level, \u0026alpha; was set at 5% and \u003cem\u003eP\u003c/em\u003e-values of less than 0.05 were considered as significant. All the data were analyzed by using IBM SPSS Statistics v 21. The concentration response data was analyzed by probit analysis (Finney 1971) to determine the median lethal time (LT\u003csub\u003e50\u003c/sub\u003e) values, their 95% confidence intervals (CI), slope \u0026plusmn; standard error (SE) and chi square (\u0026chi;\u003csup\u003e2\u003c/sup\u003e). LT\u003csub\u003e50\u003c/sub\u003e values were considered significantly different when their 95% CI did not overlap (Litchfield and Wilcoxon 1949; Robertson and Preisler 1992). Pearson correlation was applied to determine the coefficient of correlation and significance of correlation (\u003cem\u003eP\u003c/em\u003e\u0026lt;0.05) between the LT\u003csub\u003e50\u003c/sub\u003e values and different age groups of exposed larvae at highest tested concentration of different botanicals.\u003c/p\u003e"},{"header":"Results","content":"\u003ch2\u003eDelay in onset of 1\u003csup\u003est\u003c/sup\u003e oviposition\u003c/h2\u003e\n\u003cp\u003eThe 1\u003csup\u003est\u003c/sup\u003e oviposition started earlier in control than all other treatments. The highly significant delay in oviposition was observed in Eucalyptus leaf (U=9, \u003cem\u003eP\u003c/em\u003e=0.046) as compared to the control (Figure 1). Similarly, the 1\u003csup\u003est\u003c/sup\u003e oviposition was delayed with significant effect in Eucalyptus bark (U=9, \u003cem\u003eP\u003c/em\u003e=0. 046), Neem leaf (U=9, \u003cem\u003eP\u003c/em\u003e=0. 043), Neem bark (U=9, \u003cem\u003eP\u003c/em\u003e=0.046), Moringa leaf (U=9, \u003cem\u003eP\u003c/em\u003e=0.043) and Moringa bark (U=9, \u003cem\u003eP\u003c/em\u003e=0.046) as compared to control (Figure 1).\u003c/p\u003e\n\u003ch2\u003eFemale fecundity\u003c/h2\u003e\n\u003ch3\u003eNumber of egg rafts\u003c/h3\u003e\n\u003cp\u003eNumbers of egg rafts laid per female in Neem leaf (U=1, \u003cem\u003eP\u003c/em\u003e=0.11), Moringa leaf (U=1.5, \u003cem\u003eP\u003c/em\u003e=0.18) and Moringa bark (U=1, \u003cem\u003eP\u003c/em\u003e=0.08) did not differ significantly than that of control (Figure 2). Females exposed to botanical extracts of Eucalyptus bark (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05), Eucalyptus leaf (U=0, \u003cem\u003eP\u003c/em\u003e=0.05) and Neem bark (U=0, \u003cem\u003eP\u003c/em\u003e=0.05) laid similar number of egg rafts significantly lower as compared with that of control.\u003c/p\u003e\n\u003ch3\u003eNumber of eggs/female\u003c/h3\u003e\n\u003cp\u003eNumber of eggs laid by a female were significantly lower in Eucalyptus leaf (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05), Eucalyptus bark (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05), Neem leaf (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05), Neem bark (U=0, \u003cem\u003eP\u003c/em\u003e=0.05), Moringa leaf (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05) and Moringa bark (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05) compared to control (Figure 3).\u003c/p\u003e\n\u003ch2\u003eLarval emergence\u003c/h2\u003e\n\u003cp\u003eThe larval emergence was maximum in the control and was significantly reduced in Neem leaf (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05), Eucalyptus bark (U=0, \u003cem\u003eP\u003c/em\u003e=0.05), Eucalyptus leaf (U=0, \u003cem\u003eP\u003c/em\u003e=0.05) and in Moringa leaf (U=0, \u003cem\u003eP\u003c/em\u003e=0.05). However, Neem bark (U=0, \u003cem\u003eP\u003c/em\u003e=0.05) and Moringa bark (U=5, \u003cem\u003eP\u003c/em\u003e=0.83) did not show any negative effect on larval emergence as compared to control (Figure 4).\u003c/p\u003e\n\u003ch2\u003ePupation\u003c/h2\u003e\n\u003cp\u003eHighest pupation (%) in control was observed while no pupae were formed in Eucalyptus leaf (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.04) and Eucalyptus bark (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.04). Pupation (%) was significantly lower in Neem leaf (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.04) and Neem bark (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05) as compared with control. A relatively higher proportion of pupae were formed in Moringa leaf (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05) and Moringa bark (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05) but significantly lower as compared to control (Figure 5)\u003cem\u003e.\u003c/em\u003e\u003c/p\u003e\n\u003ch2\u003eAdult emergence\u003c/h2\u003e\n\u003cp\u003eAs no pupae were formed in Eucalyptus leaves and bark, there was no adult emergence in Eucalyptus leaf (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05) and bark (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.04). Moreover, no adults were emerged in Neem bark (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.04) (Figure 6). Adult emergence was significantly lower in Neem leaf extract (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.046) and Moringa leaf (U=0.0, \u003cem\u003eP\u003c/em\u003e=0.05) as compared to the control. However, Moringa bark (U=2.0, \u003cem\u003eP\u003c/em\u003e=0.28) did not reduce adult emergence of \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e as compared with control (Figure 6).\u003c/p\u003e\n\u003ch2\u003eLethal time (LT\u003csub\u003e50\u003c/sub\u003e) of botanical extracts to different ages of larvae\u003c/h2\u003e\n\u003cp\u003eIn case of 24h old larvae, the LT\u003csub\u003e50\u003c/sub\u003e values of the Neem leaf (100,000 PPM), Neem Bark (100,000 PPM), Moringa leaf (100,000 PPM), Eucalyptus leaf (100,000 PPM) were similar to each other based on the overlapping 95%CI and were the least time taking among all tested concentrations to kill the 50% population of the exposed larvae (Table 1). While, Neem leaf (100 PPM), Moringa leaf (100 PPM) and Eucalyptus leaf (100 PPM) were longest time taking concentrations among all the tested concentrations against 24h old larvae to kill the 50% population of the exposed larvae. Neem bark (100 PPM) and Moringa bark (100 PPM) did not kill any of the exposed population.\u003c/p\u003e\n\u003cp\u003eWhen 48h larvae were exposed to different botanical concentrations, Neem leaf (100,000 PPM), Neem bark (100,000 PPM), Eucalyptus Leaf (100,000 PPM) and Eucalyptus bark were the concentrations having similar LT\u003csub\u003e50\u003c/sub\u003e based on overlapping 95% CI and the shortest time taking (Table 2). Moreover, the neem leaf (100 PPM) was the longest time taking concentration among all tested concentrations to kill fifty percent of the population. However, Moringa bark (100 PPM) did not kill any larvae exposed to it.\u003c/p\u003e\n\u003cp\u003eIn case of 72h old larvae, Neem leaf (100,000 PPM), Neem bark (100,000 PPM), Moringa Bark (100,000 PPM), Eucalyptus Leaf (100,000 PPM) and Eucalyptus bark (100,000 PPM) were similar based on overlapping 95% CI and lowest time taking concentrations among all the tested concentrations (Table 3). No mortality was observed at 100 PPM of all the tested botanicals and 1000 PPM of the Neem leaf, Moringa leaf, Moringa bark and Eucalyptus Bark.\u003c/p\u003e\n\u003cp\u003eAt highest tested concentration of every botanical, LT\u003csub\u003e50\u003c/sub\u003e was highly age dependent that is higher for elder larvae and vice versa (Table 4). Our results showed a strong positive but non-significant correlation between the different age groups and LT\u003csub\u003e50\u003c/sub\u003e values of the highest tested concentrations of the tested botanicals.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eApplication of synthetic pesticides for the control of insect vectors is questioned due to resistance, environmental pollution and public health problems (Rose 2001). Whereas, various studies suggested botanical extracts are an effective tool for the control of insect pests (Mohan et al. 2006; Rahuman et al. 2008; Shahi et al. 2010). Demand for exploration of the eco-friendly control tactics against mosquitos is on rise day by day (Benelli et al. 2016). Some phytochemicals interfere with growth and development either with reproduction or acting as olfactory stimulus in form of repellent or attractant, while some act as common toxicants against larval and adult stages of mosquitoes (Shaalan et al. 2005).\u003c/p\u003e\n\u003cp\u003eOur results suggest that onset of 1\u003csup\u003est\u003c/sup\u003e oviposition was delayed as compared with control, when females were exposed to each of tested botanical extracts. Previously, Rajkumar and Jebanesan (2009) have reported negative impact of \u003cem\u003eCassia obtusifolia\u003c/em\u003e on oviposition behavior of the mosquitoes. Mehra and Hiradhar (2002) reported that \u003cem\u003eCuscuta hyaline\u003c/em\u003e crude extract showed good oviposition deterrence against \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e females. Moreover, Coria et al. (2008) also reported that \u003cem\u003eMelia azidarach\u003c/em\u003e leaf extracts has deterrent effects on oviposition of \u003cem\u003eAe. aegypti\u003c/em\u003e. Our results are also in line with that of Govindarajan et al. (2011b) who has shown that the leaf extracts of \u003cem\u003eCaesalpinia pulcherrima\u003c/em\u003e and \u003cem\u003eErvatamia coronariaand\u003c/em\u003e have activities (larvicidal and ovicidal) against \u003cem\u003eAnopheles stephensi, Ae. aegypti, and Cx. quinquefasciatus\u003c/em\u003e. Similar findings has been reported by Elango et al. (2009) that indigenous plant extracts such as \u003cem\u003eAegle marmelos, Andrographis \u003c/em\u003elineate, \u003cem\u003eAndrographis paniculata, Cocculus hirsutus, Eclipta prostrata\u003c/em\u003e, and \u003cem\u003eTagetes erecta\u003c/em\u003e have oviposition-deterrent, ovicidal, and repellent activity against \u003cem\u003eAn. subpictus\u003c/em\u003e. As oviposition was significantly delayed in exposed females in all treatments, oviposition deterrent activity of plant extracts might induce physiological changes leading to behavioral avoidance in female adult mosquitoes. It might also be due to volatile chemicals released from respective plant water extracts, which were added in their oviposition substrates. Tawatsin et al. (2006) has also reported that the relatively high oviposition deterrent activity was induced by essential oils of \u003cem\u003eCurcuma longa, Zingiber officinale\u003c/em\u003e, \u003cem\u003eVitex\u003c/em\u003e \u003cem\u003etrifolia, Melaleuca cajuputi\u003c/em\u003e, \u003cem\u003eManglietia garrettii\u003c/em\u003e, and \u003cem\u003eHouttuynia cordata\u003c/em\u003e. The delay in oviposition leads to lower female fecundity and thus it is obvious that with vast biodiversity, many bioactive compounds could be isolated and utilized for mosquito control especially \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e. Moreover, adding simple water extracts of potential botanicals in the standing waters in ditches, ponds or in sewerage lines, which are the main breeding sites of \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e, may help to deter gravid females or delaying the oviposition leading to less female fecundity and ultimately less population of \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003eLarval emergence was significantly lower in Neem leaf, Eucalyptus leaf, Eucalyptus bark and Neem bark. Prathibha et al. (2014) found that the ethyl acetate extracts of \u003cem\u003eS. mauritiana\u003c/em\u003e have marked effect results in zero hatchability at 100 ppm against \u003cem\u003eAn. stephensi\u003c/em\u003e, \u003cem\u003eAe. aegypti\u003c/em\u003e, and \u003cem\u003eCx. quinquefasciatus.\u003c/em\u003e Previously,\u003cem\u003e M. oleifera \u003c/em\u003eseed water extract has been reported to exhibit 80%, 41.3% and 9.33% larval mortality at 5%, 2.5% and 1% concentrations on larvae of \u003cem\u003eAnopheles gambiae\u003c/em\u003e (NJOM et al. 2011)\u003cem\u003e.\u003c/em\u003e At higher concentration (5% and 2.5%), no larvae reached to pupal stage but at lower concentration even few adults were emerged. Similarly,Tandon and Sirohi (2010) reported that \u003cem\u003eA. indica\u003c/em\u003e aqueous crude leaf extract caused 70-99% larval mortality followed by \u003cem\u003eGymnema sylvestre\u003c/em\u003e 44-89%, \u003cem\u003eNerium indicum\u003c/em\u003e 41-74% and \u003cem\u003eDatura metel\u003c/em\u003e 19-54% mortality of larvae. An inverse impact of botanical extracts on hatchability of eggs was observed against \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e (Govindarajan et al. 2011a). Hence, it can be concluded that unprocessed water extracts of tested plant have more potential to suppress the life table of target insects due to presence of more diverse compounds (Naumann and Isman 1995).\u003c/p\u003e\n\u003cp\u003eIn the present study, no pupation occurred in Eucalyptus leaf and in Eucalyptus bark, while very few pupae were formed in Neem leaf and Neem bark. These results are similar to NJOM et al. (2011) where higher concentration of Moringa seed extracts did not allow any pupation. Previously, hexane based leaves extract of Eucalyptus has shown potential to suppress the pupal formation in from larvae of different exposed mosquito species including \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e (Singh et al. 2007). This also indicates that plant extracts can kill the larvae as well as pupae of mosquitoes.\u003c/p\u003e\n\u003cp\u003eIn the present study, no adult emergence in Eucalyptus leaf, Eucalyptus bark and Neem bark while was highly significantly lower in Neem leaf. Elimam et al. (2009) reported that aqueous extracts from \u003cem\u003eRicinus communis\u003c/em\u003e inhibit adult emergence and also showed larvicidal as well as oviposition deterrent effect against \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e and \u003cem\u003eAn. arabiensis.\u003c/em\u003e\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThe water based botanical extracts of bark and leaves of the all tested species showed a variable range of larvicidal activity against the larvae of the \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e but have potential to be used as the traditional control agents\u003cem\u003e.\u003c/em\u003e Among the tested botanicals, mortality of the larvae was highly dose dependent. Highest mortality at higher doses and lowest or no mortality was observed at lowest doses of the tested botanicals in our study. Previously, Govindarajan et al. (2011b) reported the similar dose dependent response of the \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e to botanicals. Cetin et al. (2006) reported a similar dose dependent response of \u003cem\u003eCx. pipens\u003c/em\u003e exposed to plant extracts. Therefore, application, keeping in view the dose response of the target species, of these easily made botanical extracts shall be considered at least at small scale level, e.g., homes and poultry farms.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cul\u003e\n\u003cli\u003e\u003cstrong\u003eEthics approval and consent to participate: \u003c/strong\u003eNo human beings were subject of experiments; therefore, no consent to participate was required.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eConsent for publication: \u003c/strong\u003eNo human beings were subject of experiments; therefore, no consent to Publish was required\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003e Availability of data and material:\u003c/strong\u003e Data will be available on demand\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003e Author Contribution:\u003c/strong\u003e MK and RMS performed the experiment, SA, SM, SA and MB designed the experiment, provided materials and guidelines to perform the experiments. MK and RMS wrote the MS. SA, SM, SA and MB critically reviewed and finalized it.\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003e Funding:\u003c/strong\u003e The authors did not receive support from any organization for the submitted work\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003e Competing interests:\u003c/strong\u003e The authors hereby declared no competing interests\u003c/li\u003e\n\u003cli\u003e\u003cstrong\u003eAcknowledgements: \u003c/strong\u003eAll the authors are cordially thankful to laboratory staff in helping to perform labor work during experimentation.\u003c/li\u003e\n\u003c/ul\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eArunpandiyan G (2011) Toxicity of aqueous crude neem leaf extract against \u003cem\u003eCulex\u003c/em\u003e Int J Pharm Biomed Sci 2(1):1-3\u003c/li\u003e\n\u003cli\u003eAshfaq M, Ashfaq U (2012) Evaluation of mosquitocidal activity of water extract of Moringa oleifera seeds against \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e (Diptera: Culicidae) in Pakistan. Pak Entomol 34(1):21-26\u003c/li\u003e\n\u003cli\u003eBenelli G, Jeffries CL, Walker T (2016) Biological control of mosquito vectors: past, present, and future. Insects 7(4):52\u003c/li\u003e\n\u003cli\u003eBorah R, Kalita M, Kar A, Talukdar A (2010) Larvicidal efficacy of Toddalia asiatica (Linn.) Lam against two mosquito vectors\u003cem\u003e Aedes aegypti \u003c/em\u003eand\u003cem\u003e Culex quinquefasciatus\u003c/em\u003e. African Journal of Biotechnology 9(17):2527-2530\u003c/li\u003e\n\u003cli\u003eCetin H, Cinbilgel I, Yanikoglu A, Gokceoglu M (2006) Larvicidal activity of some Labiatae (Lamiaceae) plant extracts from Turkey. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives 20(12):1088-1090\u003c/li\u003e\n\u003cli\u003eCetin H, Erler F, Yanikoglu A (2004) Larvicidal activity of a botanical natural product, AkseBio2, against \u003cem\u003eCulex pipiens\u003c/em\u003e. Fitoterapia 75(7):724-728\u003c/li\u003e\n\u003cli\u003eCoria C, et al. (2008) Larvicide and oviposition deterrent effects of fruit and leaf extracts from \u003cem\u003eMelia azedarach\u003c/em\u003e on \u003cem\u003eAedes aegypti \u003c/em\u003e(L.)(Diptera: Culicidae). Bioresource Technology 99(8):3066-3070\u003c/li\u003e\n\u003cli\u003eElango G, Rahuman AA, Bagavan A, Kamaraj C, Zahir AA, Venkatesan C (2009) Laboratory study on larvicidal activity of indigenous plant extracts against \u003cem\u003eAnopheles subpictus\u003c/em\u003e and \u003cem\u003eCulex tritaeniorhynchus\u003c/em\u003e. Parasitol Res 104(6):1381-1388\u003c/li\u003e\n\u003cli\u003eElimam AM, Elmalik KH, Ali FS (2009) Larvicidal, adult emergence inhibition and oviposition deterrent effects of foliage extract from\u003cem\u003e Ricinus communis\u003c/em\u003e against \u003cem\u003eAnopheles arabiensis\u003c/em\u003e and \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e in Sudan. Trop Biomed 26(2):130-139\u003c/li\u003e\n\u003cli\u003eFinney D (1971) A statistical treatment of the sigmoid response curve. Probit analysis, 3rd edn Cambridge University Press, London 333\u003c/li\u003e\n\u003cli\u003eGovindarajan M, Mathivanan T, Elumalai K, Krishnappa K, Anandan A (2011a) Mosquito larvicidal, ovicidal, and repellent properties of botanical extracts against \u003cem\u003eAnopheles stephensi\u003c/em\u003e, \u003cem\u003eAedes aegypti\u003c/em\u003e, and \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e (Diptera: Culicidae). Parasitology Research 109(2):353-367\u003c/li\u003e\n\u003cli\u003eGovindarajan M, Mathivanan T, Elumalai K, Krishnappa K, Anandan A (2011b) Ovicidal and repellent activities of botanical extracts against \u003cem\u003eCulex quinquefasciatus, Aedes aegypti\u003c/em\u003e and \u003cem\u003eAnopheles stephensi \u003c/em\u003e(Diptera: Culicidae). Asian Pacific Journal of Tropical Biomedicine 1(1):43-48\u003c/li\u003e\n\u003cli\u003eGuedes DR, et al. (2017) Zika virus replication in the mosquito \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e in Brazil. Emerging Microbes \u0026amp; Infections 6(1):1-11\u003c/li\u003e\n\u003cli\u003eGuessan R, Corbel V, Akogb\u0026eacute;to M, Rowland M (2007) Reduced efficacy of insecticide-treated nets and indoor residual spraying for malaria control in pyrethroid resistance area, Benin. Emerging Infect Dis 13(2):199\u003c/li\u003e\n\u003cli\u003eHafeez F, Akram W, Shaalan EA-S (2011) Mosquito larvicidal activity of citrus limonoids against \u003cem\u003eAedes albopictus\u003c/em\u003e. Parasitol Res 109(1):221-229\u003c/li\u003e\n\u003cli\u003eKilpatrick AM, Kramer LD, Jones MJ, Marra PP, Daszak P (2006) West Nile virus epidemics in North America are driven by shifts in mosquito feeding behavior. PLoS Biol 4(4):e82\u003c/li\u003e\n\u003cli\u003eLitchfield JT, Wilcoxon F (1949) A simplified method of evaluating dose-effect experiments. J Pharmacol Exp Ther 96(2):99-113\u003c/li\u003e\n\u003cli\u003eLucantoni L, et al. (2006) Effects of a neem extract on blood feeding, oviposition and oocyte ultrastructure in Anopheles stephensi Liston (Diptera: Culicidae). Tissue and Cell 38(6):361-371\u003c/li\u003e\n\u003cli\u003eMehra B, Hiradhar P (2002) Cuscuta hyalina Roth., an insect development inhibitor against common house mosquito \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e J Environ Biol 23(3):335-339\u003c/li\u003e\n\u003cli\u003eMohan L, Sharma P, Srivastava C (2006) Evaluation of \u003cem\u003eSolanum xanthocarpum\u003c/em\u003e extract as a synergist for cypermethrin against larvae of the filarial vector \u003cem\u003eCulex quinquefasciatus \u003c/em\u003e(Say). Entomological Research 36(4):220-225\u003c/li\u003e\n\u003cli\u003eNaumann K, Isman MB (1995) Evaluation of neem \u003cem\u003eAzadirachta indica \u003c/em\u003eseed extracts and oils as oviposition deterrents to noctuid moths. Entomologia experimentalis et applicata 76(2):115-120\u003c/li\u003e\n\u003cli\u003eNJOM VS, UMEH EDN, EZE CS (2011) MALARIA VECTOR \u003cem\u003eAnopheles gambiae\u003c/em\u003e. Animal Research International 8(2):1392-1397\u003c/li\u003e\n\u003cli\u003eOkumu FO, Moore SJ, Okumu F, Moore S (2011) Combining indoor residual spraying and insecticide-treated nets for malaria control in Africa: a review of possible outcomes and an outline of suggestions for the future. Malaria journal 10(208):1-13\u003c/li\u003e\n\u003cli\u003ePrathibha K, Raghavendra B, Vijayan V (2014) Larvicidal, ovicidal, and oviposition-deterrent activities of four plant extracts against three mosquito species. Environmental Science and Pollution Research 21(10):6736-6743\u003c/li\u003e\n\u003cli\u003eRahuman AA, Gopalakrishnan G, Venkatesan P, Geetha K (2008) Larvicidal activity of some Euphorbiaceae plant extracts against \u003cem\u003eAedes aegypti \u003c/em\u003eand \u003cem\u003eCulex quinquefasciatus \u003c/em\u003e(Diptera: Culicidae). Parasitol Res 102(5):867-873\u003c/li\u003e\n\u003cli\u003eRajkumar S, Jebanesan A (2009) Larvicidal and oviposition activity of \u003cem\u003eCassia obtusifolia\u003c/em\u003e Linn (Family: Leguminosae) leaf extract against malarial vector, \u003cem\u003eAnopheles stephensi\u003c/em\u003e Liston (Diptera: Culicidae). Parasitol Res 104(2):337-340\u003c/li\u003e\n\u003cli\u003eRobertson J, Preisler H (1992) Pesticide Bioassays with Arthropods. CRC, Boca Raton, FL\u003c/li\u003e\n\u003cli\u003eRose RI (2001) Pesticides and public health: integrated methods of mosquito management. Emerging Infect Dis 7(1):17\u003c/li\u003e\n\u003cli\u003eSangshetti JN, Shinde DB, Kulkarni A, Arote R (2017) Two decades of antifilarial drug discovery: a review. RSC advances 7(33):20628-20666\u003c/li\u003e\n\u003cli\u003eShaalan EA-S, Canyon D, Younes MWF, Abdel-Wahab H, Mansour A-H (2005) A review of botanical phytochemicals with mosquitocidal potential. Environ Int 31(8):1149-1166\u003c/li\u003e\n\u003cli\u003eShah RM, et al. (2017) Larval Habitat Substrates Could Affect the Biology and Vectorial Capacity of Culex quinquefasciatus (Diptera: Culicidae). J Med Entomol 54(3):638-645\u003c/li\u003e\n\u003cli\u003eShahi M, Hanafi-Bojd A, Iranshahi M, Vatandoost H, Hanafi-Bojd M (2010) Larvicidal efficacy of latex and extract of \u003cem\u003eCalotropis procera \u003c/em\u003e(Gentianales: Asclepiadaceae) against \u003cem\u003eCulex quinquefasciatus \u003c/em\u003eand \u003cem\u003eAnopheles stephensi\u003c/em\u003e (Diptera: Culicidae). J Vector Borne Dis 47(3):185-188\u003c/li\u003e\n\u003cli\u003eSimonsen PE, Mwakitalu ME (2013) Urban lymphatic filariasis. Parasitol Res 112(1):35-44\u003c/li\u003e\n\u003cli\u003eSingh R, Dhiman R, Mittal P (2007) Studies on mosquito larvicidal properties of \u003cem\u003eEucalyptus citriodora\u003c/em\u003e Hook (family-Myrtaceae). The Journal of Communicable Diseases 39(4):233-236\u003c/li\u003e\n\u003cli\u003eTahir HM, Butt A, Khan SY (2009) Response of \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e to deltamethrin in Lahore district. J Parasitol Vector Biol 1(3):19-24\u003c/li\u003e\n\u003cli\u003eTandon P, Sirohi A (2010) Assessment of larvicidal properties of aqueous extracts of four plants against \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e Jordan Journal of Biological Sciences 147(612):1-10\u003c/li\u003e\n\u003cli\u003eTawatsin A, et al. (2006) Repellency of essential oils extracted from plants in Thailand against four mosquito vectors (Diptera: Culicidae) and oviposition deterrent effects against \u003cem\u003eAedes aegypti \u003c/em\u003e(Diptera: Culicidae). Southeast Asian J Trop Med Public Health 37(5):915-31\u003c/li\u003e\n\u003cli\u003eVan Bortel W, et al. (2008) The insecticide resistance status of malaria vectors in the Mekong region. Malaria journal 7(1):102\u003c/li\u003e\n\u003cli\u003eWakil W, Ghazanfar M, Kwon Y, Ullah E, Islam S, Ali K (2014) Testing Paecilomyces lilacinus, diatomaceous earth and Azadirachta indica alone and in combination against cotton aphid (Aphis gossypii Glover)(Insecta: Homoptera: Aphididae). African Journal of Biotechnology 11(4):821-828\u003c/li\u003e\n\u003cli\u003eWorld Health Organization W (1996) Report of the WHO informal consultation on the evaluation and testing of insecticides, CTD/WHO PES/IC/ 1996; 96.1: 69.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e. \u003cstrong\u003eLethal time (LT\u003csub\u003e50\u003c/sub\u003e) values of the 24h old larvae of \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e exposed to different botanicals\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eBotanical Extract\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eConcentration (PPM)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eLT\u003csup\u003ea1\u003c/sup\u003e\u003csub\u003e50\u003c/sub\u003e (95%CI\u003csup\u003eb\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eSlope (\u003c/strong\u003e\u003cstrong\u003e\u0026plusmn;SE)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003ex2\u003csup\u003ec\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eDf\u003csup\u003ed\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eP\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eNeem leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e184.312 (137.69-481.184)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.017\u0026plusmn;0.006\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1.475\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.478\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e107.164 (81.902-1636.850)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.022\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e4.529\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.104\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e69.976 (44.343-120.657)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.025\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e5.902\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.052\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e8.257 (17.252-21.968)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.016\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.089\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.956\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eNeem Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e95.819 (75.44-247.67)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.025\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e4.658\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.097\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e66.338 (61.365-71.625)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.027\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1.714\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.424\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e28.52 (20.468-34.481)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.029\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e3.098\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.212\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eMoringa leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e150.372 (120.38-715.81)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.032\u0026plusmn;0.015\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.13\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.937\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e106.39 (96.33-122.95)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.022\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1.867\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.393\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e75.232(68.402-83.20)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.020 \u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.729\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.694\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e11.16 (8.938-22.844)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.019 \u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1.37\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.504\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eMoringa Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e112.95(a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.023\u0026plusmn;0.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e4.847\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.089\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e92.88 (84.145-106.24)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.019\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e3.791\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.15\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e41.24(33.735-47.240)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.024 \u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2.743\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.254\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eEucalyptus Leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e140.39(119.35-198.78)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.024 \u0026plusmn; 0.006\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1.289\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.525\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e94.34 (86.39-106.13)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.022 \u0026plusmn; 0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e3.604\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.165\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e51.778 (46.67-56.50)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.029\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2.360\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.307\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e15.528(4.487-22.442)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.036 \u0026plusmn; 0.005\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.528\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.768\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eEucalyptus Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e191.880 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.021 \u0026plusmn; 0.014\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.435\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.805\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e94.343 (86.388-106.128)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.022 \u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e3.604\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.165\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e57.889(53.095-62.572)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.029\u0026plusmn; 0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2.02\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.364\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e34.738 (29.421-39.136)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.037\u0026plusmn;0.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.076\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.963\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026ldquo;A/a\u0026rdquo;indicates that software was unable to calculate the value, \u0026ldquo;A/a\u0026rdquo;indicates that software was unable to calculate the value, a1=lethal time in hours, CI=confidence intervals, X\u003csup\u003e2\u003c/sup\u003e=chi-square goodness of fit test, df=degree of freedom\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2. Lethal time (LT\u003csub\u003e50\u003c/sub\u003e) values of the 48h old larvae of \u003cem\u003eCulex quinquefasciatusa\u003c/em\u003e exposed to different botanicals \u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eBotanical Extract\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eConcentration (PPM)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eLT\u003csup\u003ea1\u003c/sup\u003e\u003csub\u003e50\u003c/sub\u003e (95%CI\u003csup\u003eb\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eSlope (\u003c/strong\u003e\u003cstrong\u003e\u0026plusmn;SE)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003ex2\u003csup\u003ec\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eDf\u003csup\u003ed\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eP\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eNeem leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e545.688 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.005\u0026plusmn;0.006\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.141\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.932\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e157.066 (124.926-360.985)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.024\u0026plusmn;0.009\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1.234\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.539\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e71.049 (64.155-79.284)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.019\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.759\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.684\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e50.351 (44.858-55.319)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.027\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.419\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.811\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eNeem Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e191.888(a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.021\u0026plusmn;0.014\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.435\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.805\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e169.160 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.025\u0026plusmn;0.014\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.268\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.874\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e70.149 (64.385-76.722)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.023\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.251\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.882\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e54.006 (49.271-58.496)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.031\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.725\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.696\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eMoringa leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e123.14 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.085 \u0026plusmn;0.372\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e123.14 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.085\u0026plusmn;0.372\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e88.49 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.022\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e8.512\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.014\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e47.54 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.020\u0026plusmn; 0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e9.935\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2.000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.007\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eMoringa Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e123.141 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.085\u0026plusmn;0.372\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e115.256 ( 88.925-6217.97)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.023 \u0026plusmn;0.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e3.971\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.137\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e72.400 (65.617-80.624)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.019\u0026nbsp; \u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2.426\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.297\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eEucalyptus Leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e285.803 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.011 \u0026plusmn; 0.008\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.653\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.721\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e207.524 (142.97-2516.20)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.015 \u0026plusmn; 0.007\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.405\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.817\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e63.864 (57.784-70.262)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.022\u0026nbsp; \u0026plusmn; 0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1.383\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.501\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e47.915 (12.16-68.37)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.030 \u0026plusmn; 0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e6.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.047\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eEucalyptus Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e191.88 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.021 \u0026plusmn;0.014\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.435\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.805\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e339.974(a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.007\u0026plusmn;0.005\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e.086\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.903\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e69.56 (63.327-76.703)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.021 \u0026plusmn; 0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1.963\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.375\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e45.524 (40.361-50.093)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.031\u0026plusmn; 0 .003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.292\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.864\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026ldquo;A/a\u0026rdquo;indicates that software was unable to calculate the value, \u0026ldquo;A/a\u0026rdquo;indicates that software was unable to calculate the value, a1= a1=lethal time in hours, CI=confidence intervals, X\u003csup\u003e2\u003c/sup\u003e=chi-square goodness of fit test, df=degree of freedom\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3.Lethal time (LT\u003csub\u003e50\u003c/sub\u003e) values of the 72h old larvae of \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e exposed to different botanicals\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eBotanical Extract\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eConcentration (PPM)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eLT\u003csup\u003ea1\u003c/sup\u003e\u003csub\u003e50\u003c/sub\u003e (95%CI\u003csup\u003eb\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eSlope (\u003c/strong\u003e\u003cstrong\u003e\u0026plusmn;SE)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003ex2\u003csup\u003ec\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eDf\u003csup\u003ed\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cem\u003eP\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eNeem leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e136.869 (116.511-182.905)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.018\u0026plusmn;0.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1.925\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.382\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e61.404 (40.060-83.379)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.028\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e4.761\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.093\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eNeem Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e114.102 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.114\u0026plusmn;0.913\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026lt;0.0001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e131.090 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.021\u0026plusmn;0.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e6.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.049\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e62.932 (30.964-99.006)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.029\u0026plusmn;0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e7.476\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.024\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eMoringa leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e131.09 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.021\u0026plusmn; 0.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e6.04\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.049\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e56.106 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.023\u0026plusmn; 0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e12.609\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.002\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eMoringa Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e157.18 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.014 \u0026plusmn; 0.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e7.024\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.03\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e80.014(72.948-89.394)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.020 \u0026plusmn; 0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.88\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.644\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eEucalyptus Leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e123.141 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.085 \u0026plusmn;0.372\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e169.79 (127.99-981.16)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.023 \u0026plusmn;0.011\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.843\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.656\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100.175 (81.453-210.426)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.027 \u0026plusmn;0.004\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e3.879\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.144\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e59.590(54.563-64.613)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.027 \u0026plusmn;0 .003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1.427\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.49\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd rowspan=\"4\"\u003e\n\u003cp\u003eEucalyptus Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003eA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003ea\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e1000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e123.141(a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.085\u0026plusmn;0 .372\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.001\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.999\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e10000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e102.028 (92.181-117.970)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.020\u0026plusmn; 0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e1.825\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.401\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e62.136 (56.978-67.410)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.026\u0026plusmn; 0.003\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.155\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e0.925\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026ldquo;A/a\u0026rdquo;indicates that software was unable to calculate the value, a1=lethal time in hours, CI=confidence intervals,\u003c/p\u003e\n\u003cp\u003eX\u003csup\u003e2\u003c/sup\u003e=chi-square goodness of fit test, df=degree of freedom\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4. Correlation between lethal time (LT\u003csub\u003e50\u003c/sub\u003e) values and different age groups of larvae of \u003cem\u003eCulex quinquefasciatus\u003c/em\u003e exposed to highest tested concentration of different botanicals \u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eBotanical Extracts\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eConcentration (PPM)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eLarval age (h)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eLT\u003csup\u003ea1\u003c/sup\u003e\u003csub\u003e50\u003c/sub\u003e (95%CI\u003csup\u003eb\u003c/sup\u003e)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u003cstrong\u003eCorrelation\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003eNeem Leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e24\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e8.257 (17.252-21.968)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e(r\u003csup\u003ec\u003c/sup\u003e=0.95, \u003cem\u003eP\u003c/em\u003e\u003csup\u003ed\u003c/sup\u003e=0.21)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e50.351 (44.858-55.319)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e61.404 (40.060-83.379)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003eNeem Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e24\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e28.52 (20.468-34.481)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e(r=0.96, \u003cem\u003eP\u003c/em\u003e=0.17)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e54.006 (49.271-58.496)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e62.932 (30.964-99.006)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003eMoringa leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e24\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e11.16 (8.938-22.844)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e(r=0.94, \u003cem\u003eP\u003c/em\u003e=0.22)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e47.54 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e56.106 (a)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003eMoringa Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e24\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e41.24(33.735-47.240)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e(r=0.94, \u003cem\u003eP\u003c/em\u003e=0.21)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e72.400 (65.617-80.624)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e80.014(72.948-89.394)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003eEucalyptus Leaf\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e24\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e15.528(4.487-22.442)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e(r=0.97, \u003cem\u003eP\u003c/em\u003e=0.17)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e47.915 (12.16-68.37)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e59.590(54.563-64.613)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003eEucalyptus Bark\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e100000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e24\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e34.738 (29.421-39.136)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e(r=0.99, \u003cem\u003eP\u003c/em\u003e=0.08)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e45.524 (40.361-50.093)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e72\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e62.136 (56.978-67.410)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026ldquo;a\u0026rdquo; indicate that software was unable to calculate the value, a1=lethal time in hours, CI=confidence intervals,\u003c/p\u003e\n\u003cp\u003eC=coefficient of correlation, \u003cem\u003eP\u003c/em\u003e=Significance of correlation (\u003cem\u003eP\u003c/em\u003e\u0026gt;0.05)\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Botanical Extract, Mosquito Management, Oviposition Behaviour ","lastPublishedDoi":"10.21203/rs.3.rs-143809/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-143809/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground: \u003c/strong\u003eThe \u003cem\u003eCulex quinquefasciatus \u003c/em\u003eSay is an important vector of many diseases of public health and veterinary importance. Environmental pollution, resistance development and health hazards associated with the use of synthetic insecticides have led to explore eco-friendly and cost effective management methods for mosquitoes. \u0026nbsp;\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eMethods: \u003c/strong\u003eIn our study, simple water extracts from leaves and bark of three tree species; \u003cem\u003eEucalyptus camaldulensis, Azadirachta indica \u003c/em\u003e(Neem)\u003cem\u003e, \u003c/em\u003eand \u003cem\u003eMoringa oleifera, \u003c/em\u003ewere tested\u003cem\u003e \u003c/em\u003eto explore their potential negative effects on the oviposition behavior and fitness of \u003cem\u003eCx. quinquefasciatus\u003c/em\u003e. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e Oviposition bioassays showed a significant delay in onset of 1\u003csup\u003est\u003c/sup\u003e oviposition in the all treatments as compred to control. Number of egg rafts and fecundity were significantly lower in all treatments compared to control. Larval emergence was significantly less in the all treatments as compared with the control except Moringa bark. Highest pupation was observed in control as compared to all other treatments. Importantly, no adult emerged in Eucalyptus leaf, bark and in Neem bark extracts. In case of toxicity bioassays, median lethal time (LT\u003csub\u003e50\u003c/sub\u003e) was lower at higher concentrations and higher at lower concentrations. At highest tested concentration of every botanical extract, LT\u003csub\u003e50\u003c/sub\u003e was highly age dependent that is higher for elder larvae and vice versa. \u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusions: \u003c/strong\u003eTherefore, it could be concluded that simple water extrcats, made locally, from trees may have a great potential to be incorporated in the mosquito control programs.\u003c/p\u003e","manuscriptTitle":"Water Extracts From Trees Negatively Affect Oviposition Behavior and Fitness of Southern House Mosquito, Culex Quinquefasciatus Say (Diptera: Culicidae): A Simple, Easily Adoptable and Cost-Effective Approach to Control Mosquitoes","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-01-18 15:59:46","doi":"10.21203/rs.3.rs-143809/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"3b6e86ec-8d72-4ae1-a661-e9c6b7885c22","owner":[],"postedDate":"January 18th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":1913810,"name":"Translational Medicine"}],"tags":[],"updatedAt":"2021-01-18T15:59:47+00:00","versionOfRecord":[],"versionCreatedAt":"2021-01-18 15:59:46","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-143809","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-143809","identity":"rs-143809","version":["v1"]},"buildId":"WrCJVZZCHTDjtuVLN7oU0","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}