Larvicidal potential of Nerium indicum and Thuja oriertelis extracts against malaria and Japanese encephalitis vector

Ethanolic and acetone extracts of Nerium indicum and Thuja orientelis have been studied against III instar larvae of Anopheles stephensi and Culex quinquefasciatus. Ethanolic extract of N. indicum is found more effective than its acetone extract against anopheline larvae with LC50 values of 185.99 and 148.05 ppm for former and 229.28 and 149.43 ppm for the later after 24 and 48 hrs of exposure. The acetone extract with LC50 values of 209.00 and 155.97 ppm is more effective in case of culicine larvae than its ethanolic extract with LC50 494.07 and 194.49 ppm after 24 and 48 hours of treatment. Ethanolic extract of T. orientelis is more effective against both the larval species with LC50 values of 13.10 and 9.02 ppm after 24 and 48 hours for anopheline and 22.74 and 16.72 ppm against culicine larvae. The acetone extract showed LC50 values of 200.87 and 127.53 ppm against anopheline and 69.03 and 51.14 ppm against culicine larvae. Thus ethanolic extract of T. orientelis is an ideal potential larvicide for both types of mosquito larvae.     

Effect of Pb,Cd and Cu on survival and development of Culex quinquefasciatus (Diptera: Culicidae)

ABSTRACT

The present research aimed to determine the lowest levels of three heavy metals (Pb, Cd and Cu) to which the larvae of Southern House Mosquito, Culex quinquefasciatus are susceptible in water. The study also aimed to investigate the effects of these heavy metals on the development of Cx. quinquefasciatus at concentrations set by Pakistan Environmental Protection Agency (Pak-EPA) as permissible levels for liquid industrial effluents. The 2nd instar larvae of Cx. quinquefasciatus were exposed to different concentrations of Pb, Cd and Cu and their effects on oviposition preference, egg hatching rate and larval development were studied. The LC₅₀ values of Pb, Cd and Cu were 12.6, 6.3 and 2.6?ppm, respectively. Gravid female mosquito adults deposited a significantly lower number of egg rafts in containers containing 0.50?ppm Pb or 1.0?ppm Cu in water. Each of the heavy metals in water resulted in significantly (p?<?0.05) lower egg hatching rate, prolonged time to pupation, lower pupation rate, prolonged time to adult emergence, lower adult emergence rate and higher female to male ratio. It is concluded that the 2nd instar larvae of Cx. quinquefasciatus are susceptible to Pak-EPA permissible levels of Pb, Cd and Cu in municipal and liquid industrial effluents.     

Effective method for extraction of larvicidal component from leaves of Azadirachta indica and Artemisia annua Linn

Leaves of Artemisia annua linn. and Azadirachta indica were extracted in petroleum ether and hexane respectively by different methods of extraction i.e. cold extraction, reflux extraction and soxhlet extraction. The crude extract obtained was tested against third instar larvae of Anopheles stephensi. On comparison of larval mortality of crude extract obtained by these three methods, both soxhlet and reflux extraction method showed 100% mortality at 200 ppm after 48 hr in case of A. annua. However LC50 (20 ppm) value of crude extract obtained by soxhlet extraction showed better results than reflux extraction (35 ppm) method after 72 hr. In case of A. indica, crude obtained by soxhlet showed 100% mortality (after 48 hr) at 250 ppm and LC50 of 69 ppm at 72 hr. Reflux extraction does not show any appreciable mortality even at 250 ppm concentration.     

Larvicidal potential of different solvent extracts and oleanolic acid derivative from Coccinia indica against vector mosquitoes

The mosquito larvicidal activities of different solvent extracts (chloroform, ethyl acetate, and methanol) from the leaves of Coccinia indica were studied. Among the extracts tested, the methanol extract showed the highest larvicidal activity against early fourth instar larvae of Culex quinquefasciatus (LC₅₀?=?142?mg?L^?1), Aedes aegypti (LC₅₀?=?158?mg?L^?1) and Anopheles stephensi (LC₅₀?=?161?mg?L^?1) after 24?h of exposure period. The lowest larval mortality was observed with the ethyl acetate extract with the LC₅₀ values of 927, 872, and 730?mg?L^?1 against Cx. quinquefasciatus, Ae. aegypti, and An. stephensi, respectively. The chloroform extract showed the moderate larvicidal activity against the mosquito species tested with the LC₅₀ values from 446 to 546?mg?L^?1. A new triterpene was isolated from the methanol extract using bio-assay guided separation and identified as an oleanolic acid derivative by NMR, IR and mass spectral studies. The compound showed prominent larvicidal activity against Cx. quinquefasciatus, Ae. aegypti and An. stephensi with the LC₅₀ values of 5.6, 5.0, and 4.8?mg?L^?1, respectively. The results of the present investigation offer an opportunity for developing alternatives to rather expensive and environmentally hazardous organic insecticides.     

Comparative efficacy of certain plant extracts alone and combination with profenofos against Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae)

Ethyl alcohol, acetone, and petroleum ether extracts of three plant species belonging to three different botanical families [Strychnos nux-vomica (Loganiaceae), Euphorbia lathyrus (Euphorbiaceae), and Datura stramonlum (Solanaceae)], a chemical insecticide; profenofos and their combinations were tested against second and fourth instars of Spodoptera littoralis under lab conditions. Results revealed that the ethanol extract of S. nux-vomica was the most effective among all plant extracts, where the corrected mortality% were 92, 81, 58, and 27% to 2nd instar and 89, 74, 34, and 11% to 4th instar at concentrations 0.5, 0.25, 0.125, and 0.0625%, respectively. Calculated LC₅₀'s were 0.11, 0.22, and 0.34% to 2nd instar and 0.17, 0.37, and 0.52% to 4th instar for ethanol, petroleum ether, and acetone extracts, respectively. Acetone extracts of all plants were of lower effect. The chemical insecticide profenofos displayed higher efficacy than plant extracts (LC₅₀ = 0.002 and 0.003% for S. littoralis 2nd and 4th larval instars, respectively). The co-toxicity factor reached 76 and 60 when mixing S. nux-vomica + profenofos and D. stramonlum + profenofos at ratio 1:1 against S. littoralis 2nd instar larvae, thus indicating a potentiating effect. While treatment of the 4th instar larvae by the same mixtures resulted in a co-toxicity factor below 20 at all mixing ratios indicating, only, an additive effect against this instar.     

Rotational application of bioinsecticide with deltamethrin-An antilarval measure for the control of filarial vector, Culex quinquefasciatus (Culicidae: Diptera)

A laboratory investigation was undertaken to study the cyclic usage of field recommended doses of Bacillus thuringiensis israelensis (Bti), Bacillus sphaericus (Bsp) and combination of Bti and Bsp (half the recommended dose of each) with deltamethrin 2.8 EC to attain better control of mosquito larvae. To understand the susceptibility status of the Culex quinquefasciatus larvae in the field, early fourth instar field collected larvae were exposed to the above treatments of biopesticides and were monitored for their persistency till 50% mortality was obtained. The larvae those survived in each treatment were transferred to the concentration of deltamethrin that gave 50% mortality. Observations on larval mortality after the treatment with deltamethrin were recorded after 24 h. The results revealed that Bti excels Bsp, as it recorded 54% mortality only on 17th day after application. The other salient finding of this study is LC50 of deltamethrin is sufficient to follow the biopesticides application for an effective control of Culex larvae.     

Comparative larvicidal potential of different plant parts of Withania somnifera against vector mosquitoes in the semi-arid region of Rajasthan

Larvicidal potential of the extracts from different parts viz. green and red fruits, seeds, fruit without seeds, leaves and roots of Withania somnifera in different solvents was evaluated against larvae of Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus, the important disease vectors prevalent in the semi-arid region. Experiments were carried out on late 3rd or early 4th instar larvae of these mosquitoes using standard WHO technique. 24 and 48 hr LC50 values along with their 95% confidence limits, regression equation, chi-square (chi2)/heterogeneity of the response have been determined by log probit regression analysis. The 24 hr LC50 values as observed for whole green fruits in water, methanol and petroleum ether were 350.9, 372.4, 576.9; 115.0, 197.1, 554.6; 154.9, 312.0, 1085.0 while corresponding values for red fruits were 473.5, 406.4,445.2; 94.7, 94.5, 1013.0; 241.8, 535.0, 893.3 mg l(-1) for An. stephensi, Ae. aegypti and Cx. quinquefasciatus respectively showing that methanol extracts were more effective against anophelines as compared to culicines when whole fruits were taken. The 24 hr LC50 values as observed for seeds in acetone, methanol and petroleum ether were 188.1, 777.5, 822.5; 245.5, 769.0, 1169.0; 140.3, 822.9, 778.4 and for fruit without seeds were 80.2, 97.6, 146.6; 88.4, 404.4, 1030.0; 30.0, 44.5, 54.2 mg l(-1) for the above mosquito species respectively showing that extract of fruit without seeds were most effective in petroleum ether followed by acetone and methanol extracts. However, experiments conducted with methanol extracts of leaves and roots of this plant species did not show any appreciable larvicidal activity and a 20-40% mortality was observed up to 500 mg l(-1) of the extracts. Overall larvae of anophelines were found more susceptible as compared to culicines to all the extracts tested. Petroleum ether extract of fruit without seeds was found most effective against all the mosquito species showing that active ingredient might be present in this part of the plant species. The study would be of great importance while planning vector control strategy based on alternative plant derived insecticides.     

Developmental change in aggregation,defense and escape behavior of buckmoth caterpillars,Hemileuca lucina (Saturniidae)

Summary Changes in response to attack and the tendency to aggregate were examined in the six larval instars of the buckmoth, Hemileuca lucina (Saturniidae). In response to simulation of attack by a parasitoid and of biting by a predator, early instars (I, II, and III) exhibited defensive behavior much more often than escape behaviors, whereas late instar larvae (IV, V, and VI) usually resorted to escape rather than defend themselves. The situations in which attacked larvae were most likely to stimulate other group members to respond were: second and third instar larvae thrashing in response to simulation of a parasitoid or headrearing in response to simulation of a biting predator; and fourth, fifth and sixth instar larvae dropping in response to either stimulus. An index of reaggregation indicated that first instar larvae had difficulty reaggregating; second, third and fourth instar larvae reaggregated quickly; and fifth and sixth larvae dispersed. As larvae developed, the change from predominantly defense to escape behaviors paralleled the decline in tendency to aggregate.     

Acephate-induced shortening of developmental duration and early adult emergence in a nontarget insect Drosophila melanogaster

Similar to several environmental monitoring studies, the present study used Drosophila melanogaster as a model nontarget organism to explore the interfering effects of an organophosphate (OP) insecticide acephate on insect life cycle parameters. Acephate, a common OP, is readily available in nature from agricultural sources as an environmental contaminant. Along with target pests, nontarget fruit flies also suffer exposure to such environmental chemical. To evaluate the effects of such exposure, initially, acute LC₅₀ of acephate for third instar larvae was investigated and found to be between 14 and 16 μg/ml. This information yielded the following experimental concentrations (0.5, 1.5, 2.5, 3.5, 4.5, and 5.5 μg/ml) of test chemical for evaluation of effect, if any, on the insect model. Results showed that mean larval duration of insect significantly decreased on treatment with acephate, whereas the mean pupal duration remained unaffected. Interestingly the decreasing trend was seen to persist in case of mean adult emergence, where treated flies emerged significantly earlier in comparison to controls. Thus, the study demonstrated that acephate-induced shortening of developmental time and early emergence in Drosophila melanogaster.     

The selective advantages of cannibalism in a Neotropical mosquito

We investigated the benefits of larval cannibalism in the Neotropical mosquito Trichoprosopon digitatum. The clutch size of the mosquito in the field was strongly correlated with adult female size, indicating a fitness advantage to being large. In controlled laboratory experiments, we compared the survivorship and eventual adult sizes of larvae that were given the opportunity to cannibalise conspecifics throughout their lifetimes with the survivorship and adult sizes of larvae that were prevented from cannibalising. Since the benefits of cannibalism are likely to depend on the context in which it occurs, the experiment was conducted at two levels of alternative food availability. When food availability was high most larvae survived to adulthood, females cannibalised more than males and there was no measurable advantage to cannibalism in terms of survival rate, emergence time or adult size. Larvae were significantly more cannibalistic when food availability was lower, although under these conditions no larvae survived to adulthood. Nevertheless, under low food an important fitness benefit to cannibalism was revealed: individuals which had the opportunity to cannibalise survived significantly longer as larvae than those which did not. This increased longevity is likely to provide an important advantage to mosquito larvae when they wait for the input of unpredictable food sources. Received: 7 October 1995/Accepted after revision: 13 April 1996     

Relative susceptibility of some common mosquito vector larvae to synthetic insecticidal compounds in north-western Rajasthan

Relative susceptibility of three important mosquito vector larvae viz., Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus, prevalent in the arid region was determined against four organophosphates (malathion, fenitrothion, fenthion, temephos) and three synthetic pyrethroid (alphamethrin, deltamethrin and fanvalerate) compounds. Studies were carried out on late 3rd or early 4th instar larvae of these species using standard WHO technique. Based on concentration mortality data LC50 and LC90 values along with their fiducial limits, regression equation, chi-square (chi2)/heterogeneity of the response have been determined by log probit regression analysis. LC50 values as observed for the above seven insecticides were 0.8097, 0.0398. 0.0432, 0.0035, 0.0025, 0.0092, 0.1006; 1.2370, 0.0531, 0.0655, 0.0076, 0.00004, 0.00004, 0.0046 and 1.4980, 0.0719, 0.0817, 0.0056, 0.00021, 0.00073, 0.0112 mg/l for the above three mosquito species respectively Among the four organophosphates tested temephos was the most effective followed by fenitrothion, fenthion and malathion. In general, Anopheles was found more susceptible as compared to the other two culicines to the above four compounds. The results also showed that larvae of Ae. aegypti were most susceptible followed by Cx. quinquefasciatus and An. stephensi to all the three pyrethroids tested. Among the three compounds tested alphamethrin was found to be the most toxic followed by deltamethrin while fanvalerate was the least toxic. The study would be of great importance while planning use of these insecticides for the control of different vector species in this area.     

Effect of nicotine (plant extract) on sex-ratio of Drosophila melanogaster (Meigen)

Experiments were conducted by using nicotine (plant extract) for its toxic effects on Drosophila melanogaster, LC50 estimated is 2.9552 microl/100 ml. Studies revealed that nicotine affects adult emergence of males and females (sex-ratio) of mutant form (Yellow) of Drosophila melanogaster.     

Size-assortative pairing across three developmental stages in the Zeus bug, Phoreticovelia disparata

The mechanisms underlying size-assortative pairing have received considerable attention. Typically, pairing is assumed to occur at, or just prior to, the adult phase of the life cycle. However, in many invertebrates, males commence associations with juvenile females who are more than a single moult away from sexual maturity. These species are ideal to explore the importance of reproductive and survival benefits as mechanisms driving size-assortative pairing. In the Zeus bug, Phoreticovelia disparata, adult males are found riding on juvenile (fourth and fifth instar) and adult females—a behaviour that is costly for females but has survival benefits for males. Using a combination of field collections and laboratory manipulations, we show that pairing is size-assortative both within and between female age classes and that riding males are smaller than non-riding males. In a series of mating trials, we revealed that males attempt to ride any female but that their riding success is dependent on female age. We also provide the first direct evidence of female resistance to male riding attempts in P. disparata. We propose that size-assortative pairing arises through adaptations that have evolved to minimise the potential costs of sexual conflict. We suggest that the selective pressure on males to maximise survival benefits is sufficiently high that it outweighs the reproductive benefits of discriminating against fourth instar females. Finally, given that female resistance is under direct selection in juvenile females, it is likely to be the main form of selective pressure for adult females.     

Life cycle and population dynamics of Pycnogonum litorale (Pycnogonida) in a natural habitat

A natural population of Pycnogonum litorale Ström was examined every 4 weeks over a period of 15 months and thereafter at yearly intervals for 15 years. Adult pycnogonids – mating couples and males carrying egg batches – and freshly hatched protonymphon larvae within these egg batches were found throughout the year. The second, third, and fourth instar larvae were only found from April to July, during the vegetation period of their hydroid host Clava multicornis. After metamorphosis to the fifth instar (first juvenile instar) the pycnogonids have a significantly larger proboscis than during the larval period, and they feed on the sea anemone Metridium senile. First juvenile instars were found on M. senile from May to August. Older and larger juvenile stages were found over longer time spans throughout the year, and the maximum number of successive instars shifted slowly from June to December. Freshly moulted adults occurred throughout the year. Males, which on the average are smaller, usually reach the adult stage during late autumn of the first year and females, at the end of the following spring. We conclude that in nature the development from egg to adult stage is completed within one year. Continuous reproduction and asynchronous embryonic development provide offspring throughout most of the year. The annual cycle is synchronized by the vegetation period of C. multicornis, the only host of these pycnogonid larvae in the investigated habitat, and by the arrest of growth during low winter temperatures. The low level of locomotory activity of P. litorale probably requires an environment in which both host species coexist. The abundance of C. multicornis, M. senile, and juvenile pycnogonids decreased from 1990 to 1996, maybe due to hydrographic conditions.     

Evaluation of the larvicidal efficacy of extracts from three plants and their synergistic action with propoxur against larvae of the filarial vector Culex quinquefasciatus (Say)

Petroleum ether extracts of Piper longum, Curcuma aromatica, and Ocimum basilicum were tested for their larvicidal activity against Culex quinquefasciatus, a filarial vector using WHO standard procedures. Among the three plants, extract of P. longum was found to be highly effective with LC₅₀ and LC₉₀ being 0.89 and 1.49 ppm, respectively. C. aromatica also showed significant larvicidal activity with LC₅₀ and LC₉₀ being 11.41 and 17.36 ppm, respectively. The LC₅₀ and LC₉₀ for O. baslicum were 76.67 and 200.56 ppm, respectively. Propoxur, a carbamate was also studied for its larvicidal activity, which showed LC₅₀ to be 0.09 ppm and LC₉₀ to be 0.16 ppm. Synergistic studies of all three plant extracts with propoxur revealed P. longum as a more efficient synergist with synergistic factor (SF) of 4.13 for LC₅₀ value and 1.68 for LC₉₀. Both C. aromatica and O. basilicum exhibited antagonistic activity at their respective LC₉₀ values with propoxur. At 1 : 1 ratio of the phytoextracts and propoxur, P. longum and C. aromatica showed synergism whereas O. basilicum displayed additive effect. Thus, P. longum was found to be a better larvicidal and synergistic agent at all tested ratios. Evidence indicates that mixed formulations of phytoextracts with insecticides might reduce the concentration of the chemicals to be applied for vector control.     

Short-term and long-term survival studies in Rana tigrina tadpoles with reference to methyl parathion toxicity

Toxicity of methyl parathion was studied on survival of Rana tigrina tadpoles. The LC50 values for short-term (upto 96 h) and long term studies (hatching to froglet) were determined. The LC50 values for 96 h and the total larval period were 4.36 and 1.30 ppm respectively. The toxicity of methyl parathion enchanced in relation to increase in concentration and duration. Teratological defects such as scoliosis and failure in emergence of fore limb were noticed. Safe levels were calculated for short-term (0.1 ppm) and long-term (0.03 ppm) studies. The present work stresses the neccessity of long term study especially for the developing amphibians.     

Toxicity assessment of heavy metals with Nostoc muscorum L

Heavy metals in aquatic ecosystem (Mn, Ni, Zn, Cu) were tested for their toxicity against Nostoc muscorum L., a common fresh water phytoplankton, with respect to chlorophyll, protein, total carbohydrate and starch contents along with growth (OD). Mn showed stimulatory effect up to 1.5 ppm for all the observed parameters. However, Ni, Zn and Cu (0.1, 0.2, 0.3, 0.4 and 0.5 ppm) showed adverse effects even at 0.1 ppm. Chlorophyll was most sensitive parameter followed by carbohydrate, protein and starch. Ni was most toxic to N. muscorum followed by Cu and Zn. The IC50 for Ni was 0.1 ppm for growth, chlorophyll and protein, while total carbohydrate and starch showed IC50 at 0.3 and 0.4 ppm, respectively. The IC50 for Cu was recorded at 0.2 ppm for all the parameters. Zn showed IC50 at 0.3 ppm for growth, chlorophyll, protein and starch at 0.4 ppm for carbohydrate. This study can be applied as bioassay using cyanobacteria for toxicity assessment of various industrial wastes in aquatic ecosystem.     

Nanoweapon: control of mosquito breeding using carbon-dot-silver nanohybrid as a biolarvicide

Health issues induced by mosquito illnesses highlight the need for effective control. Here, we developed an efficient carbon-dot-silver nanohybrid to control Anopheles stephensi and Culex quinquefasciatus mosquito species. The nanohybrid was synthesized using a thermal method without addition of any toxic-reducing agent. Monohybrids are found within the ranges of 2–6 nm for carbon-dot and 10–35 nm for silver nanoparticles with uniform distribution. The uniformly dispersed nanohybrid solutions show excellent larvicidal activity within the concentration range of 0.5–1.0 ppm. Morphological studies evidence the presence of strong bonds between nanohybrid and sulphur- or phosphorus-containing compounds such as proteins and DNA present in the larval body. This explains tissue damage at very low concentrations of nanohybrid. Therefore, this nanoweapon has high potential for field applications.     

The toxicity of heavy metals to embryos of the American oyster Crassostrea virginica

The acute toxicity of 11 heavy metals to embryos of the American oyster Crassostrea virginica was studied and the concentrations at which 50% of the embryos did not develop were determined. The most toxic metals and their LC₅₀ values were mercury (0.0056 ppm), silver (0.0058 ppm), copper (0.103 ppm) and zinc (0.31 ppm). Those metals that were not as toxic and their LC₅₀ values were nickel (1.18 ppm), lead (2.45 ppm) and cadmium (3.80 ppm). Those metals that were relatively non-toxic and their LC₅₀ values were arsenic (7.5 ppm), chromium (10.3 ppm) and manganese (16.0 ppm). Aluminum was non-toxic at 7.5 ppm, the highest concentration tested.     

Effect of mixtures of metals on the spotted Babylon snail (Babylonia areolata) under different temperature conditions

A study was conducted on the Babylon snail (Babylonia areolata) to examine the effects of copper (Cu), cadmium (Cd), nickel (Ni), and zinc (Zn) on different life stages of this gastropod. Metal toxicity significantly varied according to the life stage of the snail. The different LC₅₀ values obtained were 0.51, 5.49, 0.31, and 0.2 ppm for Cu, Zn, Cd, and Ni for the larval stage and 4.98, 15.19, 0.91, and 1.21 ppm at the juvenile stage and 8.54, 17.52, 1.14, and 1.44 ppm in the adult stage. Studies were also conducted on the effects of dual metal concentrations and experiments were repeated with temperature as a variable. Results demonstrated that metal toxicity values were altered depending on the metals involved in the combination as well as temperature under which the experiment was conducted.