Cytochrome P450 monooxygenase (P450) is a superfamily of enzymes that is important in metabolism of endogenous and exogenous compounds. In insects, these enzymes confer resistance to insecticides through its metabolic activities. Members of P450 from family 6 in insects are known to play a role in such function. In this study, we have isolated seven novel family 6 P450 from Aedes albopictus (Skuse) (Diptera: Culicidae), a vector of dengue and chikungunya fever. Induction profile of these seven genes was studied using several insecticides and xenobiotics. It was found that deltamethrin and permethrin did not induce expression of any genes. Another insecticide, temephos, inhibited expression of CYP6P15 for fivefold and twofold for CYP6N29, CYP6Y7, and CYP6Z18. In addition, copper II sulfate induced expression of CYP6M17 and CYP6N28 for up to sixfold. Benzothiazole (BZT), a tire leachate induced the expression of CYP6M17 by fourfold, CYP6N28 by sevenfold, but inhibited the expression of CYP6P15 for threefold and CYP6Y7 for twofold. Meanwhile, piperonyl butoxide (PBO) induced the expression CYP6N28 (twofold), while it inhibited the expression of CYP6P15 (fivefold) and CYP6Y7 (twofold). Remarkably, all seven genes were induced two- to eightfold by acetone in larval stage, but not adult stage. Expression of CYP6N28 was twofold higher, while expression of CYP6P15 was 15-fold lower in adult than larva. The other five P450s were not differentially expressed between the larvae and adult. This finding showed that acetone can be a good inducer of P450 in Ae. albopictus. On the other hand, temephos can act as good suppressor of P450, which may affect its own bioefficacy because it needs to be bioactivated by P450. To the best of our knowledge, this is the first report on acetone-inducible P450 in insects. Further study is needed to characterize the mechanisms involved in acetone induction in P450.
Natural insecticides from plant origin against mosquito vectors have been the main concern for research due to their high level of eco-safety. Control of mosquitoes in their larval stages are an ideal method since Aedes larvae are aquatic, thus it is easier to deal with them in this habitat. The present study was specifically conducted to explore the larvicidal efficacy of different plant parts of Ipomoea cairica (L.) or railway creeper crude extract obtained using two different solvents; methanol and acetone against late third-stage larvae of Aedes albopictus (Skuse) and Aedes aegypti (L.) (Diptera: Culicidae). Plant materials of I. cairica leaf, flower, and stem were segregated, airdried, powdered, and extracted using Soxhlet apparatus. Larvicidal bioassays were performed by using World Health Organization standard larval susceptibility test method for each species which were conducted separately for different concentration ranging from 10 to 450 ppm. Both acetone and methanol extracts showed 100% mortality at highest concentration tested (450 ppm) after 24 h of exposure. Results from factorial ANOVA indicated that there were significant differences in larvicidal effects between mosquito species, solvent used and plant parts (F=5.71, df=2, P<0.05). The acetone extract of I. cairica leaf showed the most effective larvicidal action in Ae. aegypti with LC50 of 101.94 ppm followed by Ae. albopictus with LC50 of 105.59 ppm compared with other fractions of I. cairica extract obtained from flower, stem, and when methanol are used as solvent. The larvae of Ae. aegypti appeared to be more susceptible to I. cairica extract with lower LC50 value compared with Ae. albopictus (F=8.83, df=1, P<0.05). Therefore, this study suggests that the acetone extract of I. cairica leaf can be considered as plant-derived insecticide for the control of Aedes mosquitoes. This study quantified the larvicidal property of I. cairica extract, providing information on lethal concentration that may have potential for a more eco-friendly Aedes mosquito control program.
Measuring wild pollinator services in agricultural production is very important in the context of sustainable management. In this study, we estimated the contribution of native pollinators to mango fruit set production of two mango cultivars Mangifera indica (L). cv. 'Sala' and 'Chok Anan'. Visitation rates of pollinators on mango flowers and number of pollen grains adhering to their bodies determined pollinator efficiency for reproductive success of the crop. Chok Anan failed to produce any fruit set in the absence of pollinators. In natural condition, we found that Sala produced 4.8% fruit set per hermaphrodite flower while Chok Anan produced 3.1% per flower. Hand pollination tremendously increased fruit set of naturally pollinated flower for Sala (>100%), but only 33% for Chok Anan. Pollinator contribution to mango fruit set was estimated at 53% of total fruit set production. Our results highlighted the importance of insect pollinations in mango production. Large size flies Eristalinus spp. and Chrysomya spp. were found to be effective pollen carriers and visited more mango flowers compared with other flower visitors.
The larval parasitoid Verticia fasciventris Malloch (Diptera: Calliphoridae) develops in the head of soldiers of the fungus-growing termite Macrotermes carbonarius (Hagen) (Isoptera: Termitidae). Morphological and behavioral changes in the host were evaluated and the termite castes and stages that were parasitized were identified. The larval emergence process is also described and possible mechanisms for the parasitoid fly's entry into the host body are discussed based on qualitative observations. Only a single larva per host was found. The mature larva pupated outside the host's body by exiting between the abdominal cerci. Parasitized soldiers possess a short and square-shaped head capsule, a pair of notably short mandibles, and a pair of 18-segmented antennae. Although parasitized soldiers were statistically less aggressive than healthy soldiers (P < 0.05), they expressed varying levels of aggression. Both minor and major soldiers can be parasitized and based on evidence from presoldiers, parasitization may begin during the precursor stages of soldiers. However, the stage at which parasitism first occurs has not been determined.
The red palm weevil (RPW), Rhynchophorus ferrugineus Olivier (Coleoptera: Dryophthoridae) is one of the most dangerous pests of major cultivated palms including coconut, oil palm, and sago. The larval stage of the weevil causes the most destruction of the palms as it completely destroys the palm cabbage. In this study, the larvae were given three different diets-coconut cabbage, oil palm cabbage, and sago stem, under laboratory conditions for food consumption and developmental time experiment. The protein profiles of the digestive systems of the larvae fed on these three diets were also determined. Although the coconut diet was the most consumed by RPW larvae compared to oil palm and sago diets, the growth rate of RPW larvae on oil palm diet was however significantly shorter than those on the coconut and sago diets: the RPW only need 1 mo and 9 d to complete the larval duration. Proteins profiling of eight 2-DE gel protein spots that range 50-20 kDa were identified by mass spectrometry sequence analysis. Based on the Matrix Science Software, the most dominant protein was cationic trypsin. However, based on the NCBI BLAST tool, aminopeptidase N was the most dominant enzyme. This finding can lead to the development of pest control strategies based on the antinutritional protease inhibitors as potential biocontrol agents. Urgent action to find effective control methods should be taken seriously as this weevil is presumed to be one of the serious pests of oil palm industry in Malaysia.
The oriental fruit fly, Bactrocera dorsalis (Handel) is one of the most destructive pests of fruits. The discovery of methyl eugenol (ME) as a potent male attractant for this species has led to its successful use in area-wide fruit fly control programs such as male annihilation. While the antenna is recognized as primarily responsible for male flies' detection of attractants such as ME, little is known of the involvement of the maxillary palp. Using behavioral assays involving males with intact and ablated antennae and maxillary palp structures, we seek to ascertain the relative involvement of the maxillary palp in the ability of the male fly to detect ME. In cage bioassays (distance of ≤40 cm from the source), >97% of unmodified males will normally show a response to ME. Here, we showed that 17.6% of males with their antennae ablated were still attracted to ME versus 75.0% of males with their palps ablated. However, none of the antennae-ablated males were able to detect ME over a distance of >100 cm. Furthermore, wind tunnel bioassays showed that maxillary palp-ablated males took a significantly longer time compared to unablated males to successfully detect and eventually feed on ME. These results suggest that although the antennae are necessary for detection of ME over longer distances, at shorter distances, both antennae and maxillary palps are also involved in detecting ME. Hence, those palps may play a larger role than previously recognized in maneuvering males toward lure sources over shorter ranges.
Viable biocontrol agents for mosquito control are quite rare, therefore improving the efficacy of existing biological agents is an important study. We need to have a better understanding of the predation-risk behavioral responses toward prey. This research examined prey choices by Toxorhynchites splendens by monitoring the behavioral responses of Aedes aegypti, Aedes albopictus, and Anopheles sinensis larvae when exposed to the predator. The results show that Tx. splendens prefers to consume Ae. aegypti larvae. The larvae exhibited different behavioral responses when Tx. splendens was present which suggest vulnerability in the presence of predators. "Thrashing" and "browsing" activities were greater in Ae. aegypti larvae. Such active and risky movements could cause vulnerability for the Ae. aegypti larvae due to increasing of water disturbance. In contrast, Ae. albopictus and An. sinensis larvae exhibited passive, low-risk behaviors, spending most of the time on the "wall" position near the edges of the container. We postulated that Ae. aegypti has less ability to perceive cues from predation and could not successfully alter its behavior to reduce risk of predation risk compared with Ae. albopictus and An. sinensis. Our results suggest that Tx. splendens is a suitable biocontrol agent in controlling dengue hemorrhagic vector, Ae. aegypti.
Numerous biological processes are governed by the biological clock. Studies using Drosophila melanogaster (L.) are valuable that could be of importance for their effective applications on rodent studies. In this study, the beneficial role of quercetin (a flavonoid) on H2O2 induced stress in D. melanogaster was investigated. D. melanogaster flies were divided into four groups (group I - control, group II - H2O2 (acute exposure), group III - quercetin, and group IV - quercetin + H2O2 treated). Negative geotaxis assay, oxidative stress indicators (protein carbonyls, thiobarbituric reactive substances [TBARS]), and antioxidants (superoxide dismutase [SOD], catalase [CAT], glutathione-S-transferase [GST], glutathione peroxidase, and reduced glutathione [GSH]) were measured at 4 h intervals over 24 h and temporal expression of heat shock protein-70 (Hsp70), Upd1 (homolog of IL-6 in Drosophila), and nitric oxide synthase (Nos) was analyzed by Western blotting. Groups II and IV showed altered biochemical rhythms (compared with controls). Decreased mesor values of negative geotaxis, SOD, CAT, GST, and GSH were noticed in H2O2, increased mesor of oxidative stress indicators (TBARS and protein carbonyl content) and a reversibility of the rhythmic characteristics were conspicuous after quercetin treatment. The expression levels of Hsp70, Upd1, and Nos were noticeably maximum at 04:00. Significant elevation of expression by H2O2 was nearly normalized by quercetin treatment. The possible mechanism by which quercetin modulates oxidant-antioxidant imbalance under oxidative stress could be ascribed to the modulation of the rhythmic properties. Our results will be helpful to understand the molecular interlink between circadian rhythm and oxidative stress mechanism.
Tapinoma indicum (Forel) (Hymenoptera: Formicidae) is a nuisance pest in Asia countries. However, studies on T. indicum are limited, especially in the field of molecular biology, to investigate the species characteristic at the molecular level. This paper aims to provide valuable genetic markers as tools with which to study the T. indicum population. In this study, a total of 143,998 microsatellite markers were developed based on the 2.61 × 106 microsatellites isolated from T. indicum genomic DNA sequences. Fifty selected microsatellite markers were amplified with varying numbers of alleles ranging from 0 to 19. Seven out of fifty microsatellite markers were characterized for polymorphism with the Hardy-Weinberg equilibrium (HWE) and linkage disequilibrium (LD) analysis. All seven microsatellite markers demonstrated a high polymorphic information content (PIC) value ranging from 0.87 to 0.93, with a mean value of 0.90. There is no evidence of scoring errors caused by stutter peaks, no large allele dropout, and no linkage disequilibrium among the seven loci; although loci Ti-Tr04, Ti-Tr09, Ti-Te04, Ti-Te13, and Ti-Pe5 showed signs of null alleles and deviation from the HWE due to excessive homozygosity. In conclusion, a significant amount of microsatellite markers was developed from the data set of next-generation sequencing, and seven of microsatellite markers were validated as informative genetic markers that can be utilized to study the T. indicum population.
Biology of Termites: A Modern Synthesis (Bignell DE, Roisin Y, Lo N, (Editors), Springer, Dordrecht, 576pp, ISBN 978-90-481-3976-7, e-ISBN 978-90-481-3977-4, DOI 10.1007/978-90-481-3977-4) was published in 2011. With the agreement of the publishers, we give a taxonomic index of the book comprising 494 termite entries, 103 entries of other multicellular animal species mentioned as associates or predators of termites, with 9 fungal, 60 protist, and 64 prokaryote identities, which are listed as termite symbionts (sensu stricto). In addition, we add descriptive authorities for living (and some fossil) termite genera and species. Higher taxonomic groupings for termites are indicated by 25 code numbers. Microorganisms (prokaryotes, protists, and fungi) are listed separately, using broad modern taxonomic affiliations from the contemporary literature of bacteriology, protozoology, and mycology.
Larval age and nutrition significantly affected the insect's physiology. These influences are important when rearing a population of vectors that is used to monitor the resistance level, in which standardized conditions are crucial for a more harmonized result. Little information has been reported on the effects of larval age and nutrition on the susceptibility of insects to insecticides, and therefore, we studied the effects on the susceptibility of Culex quinquefasciatus Say's (Diptera: Culicidae) larvae to temephos by comparing the median lethal concentration (LC50) after 24 hr between the second and fourth instar larvae and between the larvae that fed on protein-based and carbohydrate-based larval diets. The susceptibility of the larvae was significantly affected by the larval diets, as the larvae that fed on protein-based beef food and milk food demonstrated significantly higher LC50 value compared with the larvae that fed on carbohydrate-based food: lab food and yeast food. The larval diet interacted significantly with the larval age: while the second instar larvae were susceptible to temephos when supplied with carbohydrate-based food, the second and fourth instar larvae had no significant effect when supplied with protein-based diets, implying that a protein-rich environment may cause the mosquito to be less susceptible to temephos. This study suggested the importance of standardizing nutrition when rearing a vector population in order to obtain more harmonized dosage-response results in an insecticide resistance monitoring program. Future research could focus on the biochemical mechanism between the nutrition and the enzymatic activities of the vector.
Dual choice bioassays were used to evaluate the antifeedant property of essential oil and methanolic extract of Alpinia galanga (L.) (locally known as lengkuas) against two species of termites, Coptotermes gestroi (Wasmann) and Coptotermes curvignathus (Holmgren) (Isoptera: Rhinotermitidae). A 4-cm-diameter paper disc treated with A. galanga essential oil and another treated with either methanol or hexane as control were placed in a petri dish with 10 termites. Mean consumption of paper discs (miligram) treated with 2,000 ppm of essential oil by C. gestroi was 3.30 ± 0.24 mg and by C. curvignathus was 3.32 ± 0.24 mg. A. galanga essential oil showed significant difference in antifeedant effect, 2,000 ppm of A. galanga essential oil was considered to be the optimum concentration that gave maximum antifeedant effect. The essential oil composition was determined using gas chromatography-mass spectrometry. The major component of the essential oil was 1,8-cineol (61.9%). Antifeedant bioassay using 500 ppm of 1,8-cineol showed significant reduction in paper consumption by both termite species. Thus, the bioactive agent in A. galangal essential oil causing antifeeding activity was identified as 1,8-cineol. Repellent activity shows that 250 ppm of 1,8-cineol caused 50.00 ± 4.47% repellency for C. gestroi, whereas for C. curvignathus 750 ppm of 1,8-cineol was needed to cause similar repellent activity (56.67 ± 3.33%). C. curvignathus is more susceptible compare to C. gestroi in Contact Toxicity study, the lethal dose (LD50) of C. curvignathus was 945 mg/kg, whereas LD50 value for C. gestroi was 1,102 mg/kg. Hence 1,8-cineol may be developed as an alternative control against termite in sustainable agriculture practices.