Chemical Composition of Essential Oils from Natural Populations of Artemisia scoparia Collected at Different Altitudes: Antibacterial, Mosquito Repellent, and Larvicidal Effects.

The current study aimed to evaluate the presence of chemical variations in essential oils (EOs) extracted from Artemisia scoparia growing at different altitudes and to reveal their antibacterial, mosquito larvicidal, and repellent activity. The gas chromatographic-mass spectrometric analysis of A. scoparia EOs revealed that the major compounds were capillene (9.6-31.8%), methyleugenol (0.2-26.6%), ß-myrcene (1.9-21.4%), γ-terpinene (1.5-19.4%), trans-ß-caryophyllene (0.8-12.4%), and eugenol (0.1-9.1%). The EO of A. scoparia collected from the city of Attock at low elevation was the most active against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa bacteria (minimum inhibitory concentration of 156-1250 µg/mL) and showed the best mosquito larvicidal activity (LC50, 55.3 mg/L). The EOs of A. scoparia collected from the high-altitude areas of Abbottabad and Swat were the most repellent for females of Ae. aegypti and exhibited repellency for 120 min and 165 min, respectively. The results of the study reveal that different climatic conditions and altitudes have significant effects on the chemical compositions and the biological activity of essential oils extracted from the same species.

Prolonged Repellent Activity of Plant Essential Oils against Dengue Vector, Aedes aegypti.

Repellents are effective personal protective means against outdoor biting mosquitoes. Repellent formulations composed of EOs are finding increased popularity among consumers. In this study, after an initial screening of 11 essential oils (EOs) at the concentration of 33 µg/cm2, five of the most repellent EOs, Perovskia atriplicifolia, Citrus reticulata (fruit peels), C. reticulata (leaves), Mentha longifolia, and Dysphania ambrosioides were further investigated for repellent activity against Aedes aegypti mosquitoes in time span bioassays. When tested at the concentrations of 33 µg/cm2, 165 µg/cm2 and 330 µg/cm2, the EO of P. atriplicifolia showed the longest repellent effect up to 75, 90 and 135 min, respectively, which was followed by C. reticulata (peels) for 60, 90 and 120 min, M. longifolia for 45, 60 and 90 min, and C. reticulata (leaves) for 30, 45 and 75 min. Notably, the EO of P. atriplicifolia tested at the dose of 330 µg/cm2 showed complete protection for 60 min which was similar to the commercial mosquito repellent DEET. Gas chromatographic-mass spectrometric analyses of the EOs revealed camphor (19.7%), limonene (92.7%), sabinene (24.9%), carvone (82.6%), and trans-ascaridole (38.8%) as the major constituents of P. atriplicifolia, C. reticulata (peels), C. reticulata (leaves), M. longifolia, and D. ambrosioides, respectively. The results of the present study could help develop plant-based commercial repellents to protect humans from dengue mosquitoes.

Essential oils of four wild plants inhibit the blood seeking behaviour of female Aedes aegytpi.

Aedes aegypti (Diptera: Culicidae) mosquito is an important vector of many disease-causing pathogens. An effective way to escape from these mosquito-borne diseases is to prevent mosquito bites. In the current study, essential oils of Lepidium pinnatifidum, Mentha longifolia, Origanum vulgare, and Agrimonia eupatoria were evaluated for their repellent potential against Ae. aegypti females. Essential oils were extracted using steam distillation from freshly collected aerial parts of the plants and tested against 4-5 day old females of Ae. aegypti through the human bait technique for repellency and repellent longevity assays. The chemical composition of extracted essential oils was explored by gas chromatography coupled with mass spectrometry (GC-MS). The essential oils of L. pinnatifidum, M. longifolia, O. vulgare, and A. eupatoria at a dose of 33 µg/cm2 showed 100%, 94%, 87%, and 83% mosquito repellent activity, respectively. Furthermore, M. longifolia and O. vulgare essential oils exhibited 100% repellency at a dose of 165 µg/cm2, whereas A. eupatoria essential oil showed 100% repellency only at 330 µg/cm2. In the time-span bioassay, M. longifolia and O. vulgare essential oils showed protection against Ae. aegypti bites for 90 and 75 min, respectively whereas both A. eupatoria and L. pinnatifidum were found active for 45 min. Phenylacetonitrile (94%), piperitone oxide (34%), carvacrol (20%) and α-pinene (62%) were the most abundant compounds in L. pinnatifidum, M. longifolia, O. vulgare and A. eupatoria essential oils, respectively. The current study demonstrates that M. longifolia and O. vulgare essential oils possess the potential to be used as an alternative to synthetic chemicals to protect humans from mosquito bites.

Toxicities and Cross-Resistance of Imidacloprid, Acetamiprid, Emamectin Benzoate, Spirotetramat, and Indoxacarb in Field Populations of Culex quinquefasciatus (Diptera: Culicidae).

Culex quinquefasciatus is a major vector of several pathogens and is capable of breeding in various aquatic habitats. The extensive and injudicious use of synthetic chemicals against the mosquito species has led to the problem of insecticide resistance. To explore this resistance in detail, toxicity bioassays of imidacloprid, acetamiprid, emamectin benzoate, spirotetramat, and indoxacarb were performed on five Cx. quinquefasciatus field populations from Pakistan in addition to a laboratory susceptible strain. Compared with the susceptible strain, results for the five Cx. quinquefasciatus field populations were as follows: susceptibility to high resistance against imidacloprid (resistance ratio (RR): 0.09-11.18), susceptibility to moderate resistance against acetamiprid (RR: 0.39-8.00), susceptibility to emamectin benzoate (RR: 0.002-0.020), susceptibility to spirotetramat (RR: 0.01-0.07), and low to high resistance against indoxacarb (RR: 3.00-118.00). Correlation analyses revealed a significant positive correlation between imidacloprid, acetamiprid, and spirotetramat median lethal concentration (LC50) values, indicating the possibility of cross-resistance. In contrast, there were no significant correlations between the LC50 values of other tested insecticides, indicating the possible absence of cross-resistance. These results can assist public health authorities, medical entomologists, and pest managers to manage the insecticide resistance of Cx. quinquefasciatus as well as the associated pollution and human health issues.

Effect of Prepartum Dietary Energy Level on Production and Reproduction in Nili Ravi Buffaloes.

The objective of this study was to evaluate the effect of prepartum dietary energy level on postpartum production and reproduction in Nili Ravi buffaloes (n = 21). The buffaloes were offered low energy (LE: 1.31 Mcal/kg DM NEL (net energy for lactation)), medium energy (ME: 1.42 Mcal/kg DM NEL) or high energy (HE: 1.54 Mcal/kg DM NEL) diet for 63 days prepartum, and received the same lactation diet (LD: 1.22 Mcal/kg DM NEL) during 14 weeks postpartum. The effects of dietary energy level and week were analyzed with Proc GLIMMIX model. Dry matter intake (DMI) was lower in buffaloes fed the LE diet compared with buffaloes fed the ME or HE diet. Calf birth weight (CBW) was higher in buffaloes fed the HE diet compared with buffaloes fed the ME or LE diet. Milk production was similar in buffaloes fed the HE, ME or LE diet within 14 weeks postpartum and throughout the lactation. Milk fat was higher in buffaloes fed the LE diet compared with buffaloes fed the ME or HE diet. Milk protein and lactose yields was high in buffaloes fed the HE diet compared with buffaloes fed the ME or LE diet. Body condition score was high in HE and was affected by diet × week interactions during pre and postpartum period. The concentrations of ß-hydroxybutyrate (BHBA) and triglycerides in serum was lowest in buffaloes fed the HE diet compared with the buffaloes fed the ME or LE diet. The buffaloes fed the HE diet had early uterine involution (UI), first estrus, short dry days, and calving interval (CI) compared with buffaloes fed the ME or LE diet. None of buffaloes fed the LE diet exhibited estrus during the first 14 weeks postpartum compared with buffaloes fed the ME or HE diet. In conclusion, prepartum feeding of high energy diet can be helpful in improving the postpartum productive and reproductive performance in Nili Ravi buffaloes.

Mosquito Repellent Potential of Carpesium abrotanoides Essential Oil and Its Main Components Against a Dengue Vector, Aedes aegypti (Diptera: Culicidae).

Disease vectoring mosquitoes are a serious threat to humans. However, till today only few mosquito repellents have been identified. The current study was conducted to evaluate the repellent potential of Carpesium abrotanoides essential oil against Aedes aegypti females by human bait technique. Essential oil was extracted by steam distillation process while the identification of chemical constituents was carried out by gas chromatography-mass spectrometry. Time span repellent bioassays of C. abrotanoides essential oil in comparison to DEET were performed at three different doses (33 µg/cm2, 165 µg/cm2, and 330 µg/cm2) under laboratory conditions. Highest repellency periods for essential oil and DEET were observed at the tested dose of 330 µg/cm2 with 315 min and 720 min, respectively. Lowest repellency period of 45 min for essential oil and 105 min for DEET was recorded at the tested dose of 33 µg/cm2. Major constituents caryophyllene (24.3%) and trans-nerolidol (12.0%) of C. abrotanoides essential oil were also evaluated as repellents at three different doses (330 µg/cm2, 165 µg/cm2, and 33 µg/cm2) against Ae. aegypti. Surprisingly, trans-nerolidol completely inhibited Ae. aegypti landings for 45 min when tested at 330 µg/cm2. However, caryophyllene did not completely inhibit Ae. aegypti landing even after immediate application at the tested dose of 330 µg/cm2. At the tested dose of 330 µg/cm2, the mixture (trans-nerolidol + caryophyllene) completely inhibited Ae. aegypti landing for 60 min indicating the synergistic effect of caryophyllene. Hence, C. abrotanoides as well as its major constituent, especially trans-nerolidol, have potential to formulate as mosquito repellent comparable of DEET.

Chemical Composition, Larvicidal and Repellent Activities of Wild Plant Essential Oils against Aedes aegypti.

Bio-degradable and eco-friendly essential oils (EOs) extracted from Mentha longifolia, Salsola imbricata, Erigeron bonariensis, E. canadensis, Ailanthus altissima, and Zanthoxylum armatum were investigated for their repellent and larvicidal potential against Aedes aegypti mosquitoes. The EOs of M. longifolia, S. imbricata, E. bonariensis, E. canadensis, A. altissima, and Z. armatum exhibited 99.0%, 96.8%, 40.2%, 41.7%, 29.1%, and 13.2% repellency against mosquitoes at a tested dose of 33.3 µg/cm2, respectively. In time span bioassays, the EOs of M. longifolia, S. imbricata, E. bonariensis, and E. canadensis showed more than 40% repellency for 60 min at a tested dose of 330 µg/cm2. Larvicidal bioassays revealed that larvae of Ae. aegypti were the most susceptible to M. longifolia (LC50, 39.3 mg/L), E. bonariensis (LC50, 26.0 mg/L), E. canadensis (LC50, 35.7 mg/L), and Z. armatum (LC50, 35.9 mg/L) EOs upon 48 h exposure. The most abundant constituents in the EOs of M. longifolia, S. imbricata, E. bonariensis, E. canadensis and A. altissima were piperitone oxide (45.5%), carvone (39.9%), matricaria ester (43.1%), (31.7%) and eugenol (24.4%), respectively. Our study demonstrates that EOs of M. longifolia, S. imbricata, E. bonariensis, and E. canadensis might be used to control Ae. aegypti mosquitoes without harming humans or the environment.

Taxonomy and distribution of agriculturally important plusiinae (Lepidoptera: Noctuidae) from southern Punjab, Pakistan.

Taxonomic identification and classifications of insect pest genera Chrysodeixis and Ctenoplusia of the subfamily Plusiinae is very compulsory due to their phytophagous nature and potential to damage the cash as well as cereal crops. Taxonomy plays a key role in proper not only in identification and classification of the pest but also in designing a successful managing strategy. In current study, specimens of Chrysodeixis and Ctenoplusia genera were collected from different geographical areas of south Punjab, Pakistan and their diagnostic features were examined following taxanomic keys. The data of temperature, relative humidity and rainfall were also recorded during the study period. Genitalia was extracted by dissecting of the abdomen and inspected under Stereo microscope. The results revealed two new species, Chryodeixis maxus and Ctenoplusia oleraceaus, from south Punjab region in addition to previously reported species: Chrysodeixis furihatai. Suitable management of the voracious insect pest at appropriate time may help in sustaining the agriculture in Pakistan.

Fitness cost, realized heritability and stability of resistance to spiromesifen in house fly, Musca domestica L. (Diptera: Muscidae).

The house fly, Musca domestica L. (Diptera: Muscidae), is an insect pest of public health and veterinary importance. Spiromesifen is a new chemistry insecticide widely used for the management of sucking insect pests of vegetables and crops. In the present study, assessment of resistance risk and fitness costs associated with spiromesifen resistance in M. domestica was studied. Moreover, stability of resistance to spiromesifen and other tested insecticides (fipronil, spinosad, and bifenthrin) was evaluated in the spiromesifen-selected-strain (SPIRO-SEL-POP). After 7-generations of selection with spiromesifen, SPIRO-SEL-POP developed 108.76-fold resistance compared with the unselected strain (UNSEL-POP). The estimated value of realized heritability was 0.59 for spiromesifen resistance. Due to withdrawal of spiromesifen selection for five generations (F6-F10) on SPIRO-SEL-POP, a decline in LC50 values against spiromesifen, spinosad and bifenthrin was 0.16, 0.14 and 0.13-folds, respectively. In biological trait experiments, larval weight of Cross1 (SPIRO-SEL-POP♀ × UNSEL-POP♂) and SPIRO-SEL-POP was significantly lower than that of Cross2 (SPIRO-SEL-POP♂ × UNSEL-POP♀) and UNSEL-POP. Pupal weight of SPIRO-SEL-POP was higher when compared with Cross1 while it was similar to that of Cross2 and UNSEL-POP. Adult emergence rate of UNSEL-POP was higher than Cross1, but similar to the Cross2 and SPIRO-SEL-POP. The SPIRO-SEL-POP and Cross1 showed the lowest relative fitness when compared with USEL-POP and Cross2. Intrinsic rate of natural increase of SPIRO-SEL-POP was much lower than that of UNSEL-POP and Cross2 followed by Cross1. The SPIRO-SEL-POP exhibited lower biotic potential when compared with UNSEL-POP and Cross2 but similar to Cross1. Fecundity and hatching rates were lower in SPIRO-SEL-POP compared to UNSEL-POP. It could be concluded that spiromesifen resistance in M. domestica comes with a cost and is instable. Therefore, spiromesifen rotation with other insecticides and withdraw of its usage for some period could help to sustain its efficacy by delaying the development of resistance.

Styrene, (+)-trans-(1R,4S,5S)-4-Thujanol and Oxygenated Monoterpenes Related to Host Stress Elicit Strong Electrophysiological Responses in the Bark Beetle Ips typographus.

Bark beetles kill apparently vigorous conifers during epidemics by means of pheromone-mediated aggregation. During non-endemic conditions the beetles are limited to use trees with poor defense, like wind-thrown. To find olfactory cues that help beetles to distinguish between trees with strong or weak defense, we collected volatiles from the bark surface of healthy felled or standing Picea abies trees. Furthermore, living trees were treated with methyl jasmonate in order to induce defense responses. Volatiles were analyzed by combined gas chromatography and electroantennographic detection (GC-EAD) on Ips typographus antennae. Compounds eliciting antennal responses were characterized by single sensillum recording for identification of specific olfactory sensory neurons (OSN). Release of monoterpene hydrocarbons decreased, while oxygenated compounds increased, from spring to early summer in felled trees. In both beetle sexes particular strong EAD activity was elicited by trace amounts of terpene alcohols and ketones. 4-Thujanol gave a very strong response and the absolute configuration of the tested natural product was assigned to be (+)-trans-(1R,4S,5S)-thujanol by stereoselective synthesis and enantioselective gas chromatography. One type of OSN responded to all ketones and five other OSN were characterized by the type of compounds that elicited responses. Three new OSN classes were found. Of the eight EAD-active compounds found in methyl jasmonate-treated bark, the known anti-attractant 1,8-cineole was the one most strongly induced. Our data support the hypothesis that highly active oxygenated host volatiles could serve as positive or negative cues for host selection in I. typographus and in other bark beetles.

Characterization of inheritance and preliminary biochemical mechanisms of spirotetramat resistance in Phenacoccus solenopsis Tinsley: An economic pest from Pakistan.

Phenacoccus solenopsis is an economically important insect pest of different agronomic and horticultural field crops. In Pakistan, the cotton crop was severely attacked by P. solenopsis during 2007 and since then a varied group of insecticides are used by farmers to manage this pest. As a result, insecticide resistance has become a barrier in control of P. solenopsis. The current study was designed to explore the basics of genetics, realized heritability and possible genetic mechanisms of resistance against spirotetramat in P. solenopsis. Before selection, the wild population (Wild-Pop) showed 5.97-fold resistance when compared with lab-reared susceptible strain (Susceptible Lab-Pop). The P. solenopsis was selected with spirotetramat to 21 generations, called Spiro-SEL Pop, which showed 463.21-fold resistance as compared with the Susceptible Lab-Pop. The values of LC50 for F1 (Spiro-SEL Pop ♂ × Susceptible Lab-Pop ♀) and F1 (Spiro-SEL Pop ♀ × Susceptible Lab-Pop ♂) populations were statistically similar and values of dominance level were 0.42 and 0.54, respectively. Reciprocal crosses between Susceptible Lab-Pop and Spiro-SEL Pop showed that resistance was of autosomal in nature with incomplete dominant traits. According to the fit test, monogenic model estimation of the number of genes, which are responsible for the development of spirotetramat resistance in a population of P. solenopsis, showed that multiple genes are involved in controlling the resistance levels in tested strains of P. solenopsis. The value of heritability for resistance against spirotetramat was 0.13 in P. solenopsis. Our results suggested the presence of a metabolic-based resistance mechanism associated with the monooxygenases in P. solenopsis, while testing the synergism mechanism. These results will provide the baseline to design an effective control strategy to manage P. solenopsis in the field.

Eugenol, a Plant Volatile, Synergizes the Effect of the Thrips Attractant, Ethyl Iso-Nicotinate.

The onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), is a polyphagous pest that causes serious damage to agricultural crops, vegetables, and ornamental plants worldwide. Farmers rely on the extensive usage of synthetic chemical insecticides to control T. tabaci. There is a dire need to develop alternative control strategies to overcome the problems posed by chemical insecticides. Efficient traps would allow sensitive monitoring and possibly mass trapping. A field experiment was conducted to evaluate the potential of three plant compounds with known release rates (ranging from 6-30 mg/d); eugenol (Eug), 1, 8-cineole (eucalyptol), and linalool in all possible combinations with a thrips attractant, ethyl iso-nicotinate (EI). A combination of EI with Eug increased the effect of EI by attracting 100% more thrips (effect size, 1.95) as compared to the control of EI alone. Catches in remaining treatments were lower and or not significantly different from EI alone. The results from our study could be used to develop improved volatile blends to be used for monitoring traps. Our data suggests that these traps could be effective even at very low populations.

Influence of Frequently Used Chemical Insecticides on Mycoflora Carried by Common Housefly, Musca domestica L.

The housefly, Musca domestica L. (Diptera: Muscidae), is a major medical and veterinary insect pest. It serves as a vector of many pathogenic microorganisms causing spoilage of food and diseases in human and animals. Use of chemical insecticides is adapted as a principal tool to manage housefly. Insecticides have many unforeseen ecological consequences including effects on non-target organisms. In the present study, we have assessed the effects of 10 different synthetic insecticides on the growth of mycoflora associated with the external body of the housefly by using poison food technique. Our results reveled that all synthetic insecticides enhanced the growth. Surprisingly, in most of the cases, mycelial growth of fungi was significantly increased at high concentration as compared with lower concentration. This study provides useful information about the dangerous effects of synthetic insecticides on environment by increasing the spread of various non-target pathogenic, mycotoxigenic, and food spoiling fungi, carried by houseflies.

Functional evolution of Lepidoptera olfactory receptors revealed by deorphanization of a moth repertoire.

Insects detect their hosts or mates primarily through olfaction, and olfactory receptors (ORs) are at the core of odorant detection. Each species has evolved a unique repertoire of ORs whose functional properties are expected to meet its ecological needs, though little is known about the molecular basis of olfaction outside Diptera. Here we report a pioneer functional analysis of a large array of ORs in a lepidopteran, the herbivorous pest Spodoptera littoralis. We demonstrate that most ORs are narrowly tuned to ubiquitous plant volatiles at low, relevant odorant titres. Our phylogenetic analysis highlights a basic conservation of function within the receptor repertoire of Lepidoptera, across the expansive evolutionary radiation of different major clades. Our study provides a reference for further studies of olfactory mechanisms in Lepidoptera, a historically crucial insect order in olfactory research.

Selection, Realized Heritability, and Fitness Cost Associated With Dimethoate Resistance in a Field Population of Culex quinquefasciatus (Diptera: Culicidae).

Mosquitoes are known to be vectors of numerous diseases leading to human morbidity and mortality at large scale in the world. Insecticide resistance has become a serious concern in controlling the insect vectors of public health importance. Dimethoate is an organophosphate insecticide used to control different insect pests including mosquitoes. Biological parameters of susceptible, unselected, and dimethoate-selected strains of Culex quinquefasciatus Say were studied in the laboratory to recognize resistance development potential and associated fitness cost. The dimethoate-selected strain showed 66.48-fold resistance to dimethoate compared with the susceptible strain after three continuous selections of generations. Realized heritability estimates of dimethoate resistance in Cx. quinquefasciatus yielded a value of 0.19. In dimethoate-selected strain, the biological traits including larval weight, survival from first instar to pupae, fecundity, number of next-generation larvae, relative fitness, net reproductive rate, intrinsic rate of natural increase, and biotic potential were significantly reduced as compared with the unselected strain. However, adult longevity, mean relative growth rate, weight of egg raft, female ratio, pupal duration, and emergence rate of the dimethoate-selected strain did not differ significantly compared with that of the unselected strain. This study provides useful information to devise retrospective management strategy for dimethoate resistance in Cx. quinquefasciatus.

Larval Habitat Substrates Could Affect the Biology and Vectorial Capacity of Culex quinquefasciatus (Diptera: Culicidae).

Culex quinquefasciatus Say is an important disease vector throughout much of the world. Experiments were conducted to determine the effects of different larval habitat substrates on the fitness and biting efficiency of Cx. quinquefasciatus adults. Our findings indicate that the development time (egg to adult) of larvae reared in irrigation water was 8.63 d while that of larvae reared in distilled water was 17.10 d (Effect size = 0.95). However, the rate of adult emergence was similar for all the tested treatments. Furthermore, the mean weight of an egg raft varied between larval habitats: distilled water (1.83 mg), rainfall water (1.25 mg), irrigation water (1.52 mg), and sewerage water (2.52 mg) (Effect size = 0.91). But, the fecundity (eggs per female) and hatchability (%) were statistically similar in all the rearing mediums (Effect size = 0.79). Longevity of females in all the tested populations did not differ significantly (Effect size = 0.91). The mean relative growth rates of larvae reared in tap water (0.80) and distilled water (0.86) habitats were lower than growth rates in all other rearing habitats (Effect size = 0.96). The intrinsic rate of natural increase in tap water (0.27) and irrigation water (0.35) was significantly higher than that in distilled water (0.09) and sewerage water (0.16) (Effect size = 0.84). Adults reared in rain water had the highest biting efficiency among all the tested populations. These results provide useful information for the management of Cx. quinquefasciatus.

Assessment of resistance risk to fipronil and cross resistance to other insecticides in the Musca domestica L. (Diptera: Muscidae).

Fipronil, a phenyl-pyrazole insecticide has been used frequently for the control of disease vector house flies, Musca domestica L., (Diptera: Muscidae) worldwide including Pakistan. This experiment was performed to determine the selection and assessment of fipronil resistance evolution along with cross resistance to other three insecticides. After 26 generations of selection, the house fly strain developed 430-fold resistance to fipronil compared to a susceptible strain. Realized heritability (h(2)) of resistance to fipronil was 0.05. The projected rate of resistance development revealed that if 30-90% house flies were selected then a tenfold increase in lethal concentration 50 happened after 95.51-26.59 generations for fipronil (h(2)=0.05, Slope=2.34). At similar slope, if h(2)=0.15, then 31.84-8.86 generations are required for tenfold increase in LC50 at 30-90% selection intensity, respectively. Likewise, if h(2)=0.25, then similar would occur in 19.10-5.32 generations. Differences in any of the variable would affect the rate of resistance development. Selection with fipronil did not increase the level of resistance to lambda-cyhalothrin, profenofos and indoxacarb, suggesting no cross resistance to these insecticides. The results of our study concluded that house flies have the potential to develop resistances following continued selection pressure with fipronil.

Effects of different animal manures on attraction and reproductive behaviors of common house fly, Musca domestica L.

Insects rely mainly on their well-developed and highly sophisticated olfactory system to discriminate volatile cues released from host and nonhost substances, mates, oviposition substrates, and food sources. Onset of first mating, mating duration, and onset of first oviposition, oviposition period, fecundity (number of eggs laid by a female), and longevity of freshly emerged Musca domestica L. (Diptera: Muscidae) adults were observed in the presence of different animal manures: cow, horse, donkey, poultry, and an artificial diet. The M. domestica adults exposed to horse manure showed a delay in onset of first mating and first oviposition, prolonged mating duration, and reduced fecundity compared to the artificial diet (control). Likewise, the fecundity was reduced in the presence of donkey manure as compared to artificial diet. The onset of first mating was delayed and duration of first mating was shortened in the presence of cow manure as compared to artificial diet and no oviposition was observed throughout the duration of the experiment. However, the reproductive behaviors and all fitness measures in adults exposed to poultry manure were similar or even better, compared to the artificial diet. Surprisingly, in a free-choice attraction assay, the highest numbers of adult flies were attracted toward the cow manure as compared to all other manures as well as the artificial diet. However, the numbers of flies captured in all other types of manures were not different than the artificial diet (control). Furthermore, chemical analysis of headspace samples of manures revealed qualitative differences in odor (volatile) profiles of all manures and artificial diet, indicating that behavioral differences could be due to the differences in the volatile chemistry of the adult ovipositional substrates and larval growth mediums. This study may contribute toward both understanding the linkage between ecological adaptations and host selection mechanisms and the development of pest management strategies against this serious pest of medical and veterinary importance.

Concurrent modulation of neuronal and behavioural olfactory responses to sex and host plant cues in a male moth.

Mating has profound effects on animal physiology and behaviour, not only in females but also in males, which we show here for olfactory responses. In cotton leafworm moths, Spodoptera littoralis, odour-mediated attraction to sex pheromone and plant volatiles are modulated after mating, producing a behavioural response that matches the physiological condition of the male insect. Unmated males are attracted by upwind flight to sex pheromone released by calling females, as well as to volatiles of lilac flowers and green leaves of the host plant cotton, signalling adult food and mating sites, respectively. Mating temporarily abolishes male attraction to females and host plant odour, but does not diminish attraction to flowers. This behavioural modulation is correlated with a response modulation in the olfactory system, as shown by electro-physiological recordings from antennae and by functional imaging of the antennal lobe, using natural odours and synthetic compounds. An effect of mating on the olfactory responses to pheromone and cotton plant volatiles but not to lilac flowers indicates the presence of functionally independent neural circuits within the olfactory system. Our results indicate that these circuits interconnect and weigh perception of social and habitat odour signals to generate appropriate behavioural responses according to mating state.

Identification of plant semiochemicals and characterization of new olfactory sensory neuron types in a polyphagous pest moth, Spodoptera littoralis.

Phytophagous insects use blends of volatiles released from plants to select hosts for feeding and oviposition. To behaviorally analyze complex blends, we need efficient and selective methods for elucidating neuron types, their ligands, and specificity. Gas chromatography-combined single sensillum recordings (GC-SSRs) from antennal olfactory sensilla of female moth, Spodoptera littoralis revealed 38 physiologically active peaks in the headspace volatile blends from both larvae-damaged cotton plants and lilac flowers. Using GC-combined mass spectrometry, 9 new physiologically active compounds were identified from damaged cotton and 11 from lilac compared with earlier electrophysiological studies using antennae of female S. littoralis. We characterized 14 novel classes of olfactory sensory neurons (OSNs). Among these, we found the first 2 ligands for a frequent type of short trichoid sensillum, for which no ligands were identified earlier. By using GC-SSR, a substantial increase in functional classes of OSNs and active compounds, 40% and 34% more, respectively, compared with recent studies using GC-electroantennogram or SSR using single compounds was detected. Compared with the estimated number of corresponding antennal olfactory receptors, the OSN classes now correspond to 83% of a likely maximum. The many specialist OSNs observed may facilitate behavioral confirmation of key plant volatiles in blends.