The tea green leafhopper, Empoasca vitis Göthe, is one of the most serious pests in tea growing areas. This study investigated the roles played by olfaction and vision in host orientation behavior. The compound eye of E. vitis was found to be a photopic eye; few olfactory sensilla were found on the antennae, while abundant gustatory sensilla were recorded on the mouthparts. Three opsin genes (EV_LWop, EV_UVop, EV_Bop) were isolated and found to be mainly expressed in the compound eye compared with other parts of the body. Immunolocalization indicated that the opsins mainly located in the different regions of rhabdom. The transcription levels of EV_LWop, EV_Bop and EV_UVop were reduced by 77.3, 70.0 and 40.0%, respectively, by RNA interference induced by being fed a special RNA-rich diet for 6 days. The rate of tropism to host color was effectively impaired by 67.6 and 29.5% in the dsEV_LWop and dsEV_Bop treatment groups, but there was no significant change in the dsEV_UVop group. The determination of the cause of the tropism indicated that odors from the host over long distances were unable to attract E. vitis and were only detected when the insects were close to the host. The developed compound eye of E. vitis plays a leading role in host location, and the long-wavelength opsin significantly affects the tropism to host color; the lack of olfactory sensilla results in long-distance odors not being able to be detected until the insect is near to the host-plant. The understanding of these behavioral mechanisms, especially the importance of opsin genes is expected to be useful for pest management.

The present study investigated the patterns of microorganisms in an artificial larval diet during Dacus ciliatus (Diptera; Tephritidae) larval development. Microbial population contents in the diet of total heterotrophic bacteria, yeast and molds, coliform and lactobacilli, and their dynamics during development, were monitored. Initially, the microbial composition in diet trays failing to produce viable pupae and in trays successfully producing pupae and adult flies was characterized. The failing diet trays contained large populations of lactobacilli that increased during larval development, and low populations of coliforms. In contrast, the successful diet showed an increasing population of coliforms and a low, or undetected, population of lactobacilli. To study the hypothesis that lactobacilli affect D. ciliatus larval development, we conducted controlled inoculation experiments in which Lactobacillus plantarum was added into fresh diet at the time of egg seeding. L. plantarum inoculated trays showed no production of D. ciliatus. Control trays without lactobacilli inoculation showed variable results. One tray successfully produced viable pupae and adults, and showed a slight and slow increase in the indigenous populations of lactobacilli. The second tray, however, failed to produce pupae and showed a fast increase of the indigenous lactobacilli to very high levels. Monitored pH trends in L. plantarum-inoculated diet showed a sharp pH decrease during the first 4 days of larval development from 5 to less than 4 units, while successful diet, producing viable D. ciliatus pupae and adults, showed a moderate pH drop during most of the larval development period. The paper discusses the possible ecological interactions between D. ciliatus larvae, the microbial content of the diet and the physical properties of the diet. The discussion also points out at the usefulness of this approach in understanding and managing mass production parameters of tephritid fruit flies industrial diets used for Sterile Insect Technique.

Investigations on oviposition behaviour of dengue vectors are critical for effective controlling of vector breeding. Therefore, the aim of the present study was to determine the oviposition behaviour of dengue vectors, Aedes aegypti and Aedes albopictus in Sri Lanka. Batches of 1000 adult mosquitoes (1 : 1, male: female ratio) housed in rearing cages were used for each experimental setup from Ae. aegypti and Ae. albopictus. Oviposition responses with respect to the size of the ovitrap, colours of the ovitrap, water source, sodium chloride (NaCl) concentration and presence/absence of larvae were evaluated by enumerating the number of eggs laid in the ovitraps. The analysis of variance and cluster analysis were used to investigate the significance in the variations among oviposition. The number of eggs laid by both species were improved with the increasing size of ovitraps. Ae. albopictus indicated the highest mean number of eggs in 0.2% of NaCl than in the ovitraps filled with distilled water. However, the egg laying preference was reduced with increasing salinity in both species. Drain water with low dissolved oxygen (DO) level (0.43 ± 0.12 mg l−1) was the preferred water source for both species, while a significantly high oviposition rate was observed in ovitraps with larvae. Black colour ovitraps attracted the majority of gravid females, while white was least preferred. There were no significant variations among oviposition behaviours of Ae. albopictus and Ae. aegypti. The ability of these vectors to breed in waste water with low DO levels may lead them to attain wide dissemination in the natural environment, enhancing their potential threat to human life.

Anoplolepis gracilipes is one of the six most widespread and pestiferous invasive ant species. Populations of this invader in Arnhem Land, Australia have been observed to decline, but the reasons behind these declines are not known. We investigated if there is evidence of a pathogen that could be responsible for killing ant queens or affecting their reproductive output. We measured queen number per nest, fecundity and fat content of queens from A. gracilipes populations in various stages of decline or expansion. We found no significant difference in any of these variables among populations. However, 23% of queens were found to have melanized nodules, a cellular immune response, in their ovaries and fat bodies. The melanized nodules found in dissected queens are highly likely to indicate the presence of pathogens or parasites capable of infecting A. gracilipes. Queens with nodules had significantly fewer oocytes in their ovaries, but nodule presence was not associated with low ant population abundances. Although the microorganism responsible for the nodules is as yet unidentified, this is the first evidence of the presence of a pathogenic microorganism in the invasive ant A. gracilipes that may be affecting reproduction.

Dormancy can be defined as a state of suppressed development allowing insects to cope with adverse conditions and plant phenology. Among specialized herbivorous insects exploiting seasonal resources, diapause frequently evolves as a strategy to adjust to predictable plant seasonal cycles. To cope with acyclic and unpredictable climatic events, it has been found for some insects that a proportion of the population undergoes prolonged dormancy. We compared the response of three species in the Rhagoletis cingulata species group exploiting plants differing in fruiting phenology from environments varying in frequency and timing of acyclic climatic catastrophic events (frost during flowering and fruit set) and varying also in the time of the onset of the rainy season. Small proportions (<2%) of Central and Northeastern Mexico R. cingulata and Rhagoletis turpiniae, and a few associated adult parasitoids, emerged without becoming dormant. Pupae exposed to 5°C for different periods of time (0–16 weeks) became dormant and emerged as adults in proportions <30%, compared with other studied species in the group from the USA (up to 80%). Large proportions (19–55%) of exposed pupae entered prolonged dormancy (>10 months), and large proportions of pupae died without emerging as adults. The number of days elapsed from the end of artificial winter and adult eclosion was longer for R. cingulata exploiting late fruiting Prunus serotina in Northeastern Mexico than for flies recovered from earlier fruiting plants in the central Altiplano. Rhagoletis turpiniae and northeastern R. cingulata pupae suffered high proportions of parasitism. Large proportions of R. cingulata from central Mexico engaging in prolonged dormancy may be explained by the fact that flowering and fruit set for its host, P. serotina var capuli, driven by the timing of maximum precipitation, matches a period of highest probability of frost often resulting in large areas with fruitless trees at unpredictable time intervals. As a consequence of differences in host plant fruiting phenology, central and northeastern Mexican R. cingulata were found to be allochronically isolated. Prolonged dormancy may have resulted in escape from parasitism.

The brown planthopper Nilaparvata lugens Stål (BPH) can be found year-round in tropical region and causes severe damage to rice. Although there has been documented BPH damage to rice crops in the past decade in Cambodia, the extent of this epidemic is poorly understood. Here, we examined the time variation of BPH population in the abundance of morphotypes in 13 main rice-producing provinces (86 sites) by aspirator method and in the Takeo Province (five sites) by yellow sticky trap method. At least three generations were observed during the 3-month collection period in the rainy growing season. Regarding the occurrence of BPH morphotypes, in July the macropterous adults were restricted to south Cambodia and in August all morphotypes, adults (macropterous and brachypterous) and nymphs, appeared in all sampling sites. To explain the difference of regional distribution, the genetic differentiation was analyzed in south and northwest Cambodia (three sites) by using single nucleotide polymorphisms (SNP) analysis via genotyping-by-sequencing (GBS) using next-generation sequencing. The 2455 SNPs obtained by GBS clarified the three sub-populations and they corresponded to the expected dissemination patterns. These results provide a clue to understand the differentiation and epidemic of BPH in Cambodia.

We tested the sulfur-modulated plant resistance hypothesis using potted cabbage (Brassica oleracea var. capitata) plants that were grown without and with increasing levels of sulfur fertilization. Changes in plant chemical traits were assessed and developmental performance of Plutella xylostella, a highly host-specific leaf-chewing insect, was followed. Leaf sulfur concentration gradually increased with growing addition of sulfur in soil; however, there was a generalized saturation response curve, with a plateau phase, for improvements in total leaf nitrogen, defense glucosinolates and insect performance. Plutella xylostella performed better in sulfur-fertilized cabbage probably because of the higher level of nitrogen, despite of the higher content of glucosinolates, which are toxic for many non-specialized insects. Despite the importance of sulfur in plant nutrition and production, especially for Brassica crops, our results showed that sulfur fertilization could decrease plant resistance against insects with high feeding specialization.

The bean bug (Riptortus pedestris) is a pest of soybeans and other legumes in Japan and other Asian countries. It enters a facultative adult diapause on exposure to short days. While photoperiodism and diapause are well understood in R. pedestris, knowledge of cold tolerance is very limited, as is information on the effect of diapause on cold tolerance. We examined the effect of photoperiod, cold acclimation, and feeding status on cold tolerance in R. pedestris. We found that cold acclimation significantly increased survival at −10°C in both long- and short-day adult R. pedestris. Since the difference in cold survival between long- and short-day cold-acclimated groups was only marginal, we conclude that entering diapause is not crucial for R. pedestris to successfully pass through cold acclimation and become cold tolerant. We observed similar effects in 5th instar nymphs, with both long- and short-day cold-acclimated groups surviving longer cold exposures compared with non-acclimated groups. Starvation, which was tested only in adult bugs, had only a negligible and negative impact on cold survival. Although cold tolerance significantly increased with cold acclimation in adult bugs, supercooling capacity unexpectedly decreased. Our results suggest that changes in supercooling capacity as well as in water content are unrelated to cold tolerance in R. pedestris. An analysis of metabolites revealed differences between the treatments, and while several metabolites markedly increased with cold acclimation, their concentrations were too low to have a significant effect on cold tolerance.

Gall-making Cynipidae manipulate the leaves of host plant to form galls where offspring find shelter and food. The relationship between oak gallwasp and biochemical mechanisms of galls still requires a better understanding. So, in this research, protein and phenolic compound contents, as well as the activity of antioxidative enzymes and pathogenesis-related (PR) proteins were determined. Galls caused by asexual generation of Cynips quercusfolii L., Neuroterus numismalis (Fourc.) and N. quercusbaccarum L., as a model were used. All cynipid species modified the protein levels of gall tissues, but they cannot be treated as protein sinks. Significantly higher levels of phenols were observed in the galled leaves and galls of all cynipid species when compared with the control tissues. Peroxidase and polyphenol oxidase activity was usually low or showed no activity in galled tissues of all species. PR proteins, such as chitinase and β-1,3-glucanase, had a similar activity profile. Their activity significantly increased in the leaves with galls of all cynipid species, especially those infested with C. quercusfolii. Data generated in this study clearly indicate that galling Cynipidae manipulate the biochemical machinery of the galls for their own needs. However, the pattern of the biochemical features of leaves with galls and galled tissues depends on gall-making species.

A comprehensive study on digestive trypsin was undertaken in the larval midgut of Pieris brassicae L. Results of enzymatic compartmentalization showed a significantly higher activity of crude trypsin in the anterior larval midgut rather than posterior-midgut. Using Diethylaminoethyl cellulose fast flow column chromatography a purified trypsin was obtained by specific activity of 21 U mg−1 protein, recovery of 22%, purification fold of 28-fold and molecular weight of 25 kDa. This purified enzyme showed the highest activity at pH 8 and the corresponding temperature of 40°C. However, the specific inhibitors used including 4-(2-Aminoethyl) benzenesulfonyl fluroride hydrochloride, N-p-Tosyl-L-lysine methyl ester hydrochloride and Soybean Trypsin Inhibitor significantly lowered the activity of the purified enzyme in vitro. Moreover, the activity of trypsin and likewise the nutritional indices were significantly altered in the larval midgut feeding upon the leaves treated by 1 mM concentration of each inhibitor in comparison with control. Determination of enzymatic characteristics of insect trypsins is crucial in paving the path for controlling pests by potential natural compounds via transgenic plants.

Galeruca daurica (Joannis) is a new outbreak pest in the Inner Mongolia grasslands in northern China. Heat shock protein 10 and 60 (Hsp10 and Hsp60) genes of G. daurica, designated as GdHsp10 and GdHsp60, were cloned by rapid amplification of cDNA ends techniques. Sequence analysis showed that GdHsp10 and GdHsp60 encoded polypeptides of 104 and 573 amino acids, respectively. Sequence alignment and phylogenetic analysis clearly revealed that the amino acids of GdHsp10 and GdHsp60 had high homology and were clustered with other Hsp10 and Hsp60 genes in insects which are highly relative with G. daurica based on morphologic taxonomy. The mRNA expression analysis by real-time PCR revealed that GdHsp10 and GdHsp60 were expressed at all development stages and in all tissues examined, but expressed highest in eggs and in adults’ abdomen; both heat and cold stresses could induce mRNA expression of GdHsp10 and GdHsp60 in the 2nd instar larvae; the two Hsp genes were expressed from high to low with the extension of treatment time in G. daurica eggs exposed to freezing point. Overall, our study provides useful information to understand temperature stress responses of Hsp60 and Hsp10 in G. daurica, and provides a basis to further study functions of Hsp60/Hsp10 relative to thermotolerance and cold hardiness mechanism.

European dry-wood termites belong to the genus Kalotermes (Kalotermitidae), one of the two termite genera in Europe. Until the recent description of two new species, Kalotermes italicus in Italy and Kalotermes phoenicae in the eastern Mediterranean area, Kalotermes flavicollis was the only taxon known in this region. The presence of additional entities, suggested by morphological and physiological variation observed in K. flavicollis, was supported by molecular studies revealing four distinct genetic lineages: lineage A, K. flavicollis sensu strictu, from the Aegean area to Italy; lineage B, in Tuscany; lineage SC, in Sardinia and Corsica; lineage SF, in southern France. Lineages A and B may form mixed colonies, suggesting hybridization. To draw a more detailed picture of Kalotermes evolution and biogeography in Europe, we analyzed samples from previously unsampled areas, such as Spain and southern Italy, by means of the highly informative cox1/trnL/cox2 mitochondrial DNA marker. Overall, phylogenetic analyses confirmed previously identified lineages and taxa, but widened the distribution of the lineage SC to the mainland and of the lineage SF to Spain and Portugal. Results further provided evidence for the synonymy between lineage B and K. italicus. Species delimitation analysis suggested that the three K. flavicollis lineages, as well as K. italicus, can be separate taxa. Data also suggest a possible interspecific hybridization between K. italicus and both K. flavicollis lineages A and SC.

The subterranean termite Reticulitermes grassei Clément causes lesions in the trunk of Quercus suber L. by constructing feeding galleries, but no information is available regarding other Quercus species from the Mediterranean region. This work aimed to study the suitability of the other main oak species of Mediterranean forests as a food resource for R. grassei. Two experiments, choice and non-choice feeding, were conducted lasting for 15, 30, and 45 days each. In the non-choice experiment, termites were offered one of the following food types: Quercus suber, Quercus ilex L., Quercus faginea Lam, cork or Pinus pinea L., which was considered the control. The choice feeding experiment used all the same food types listed above, supplied simultaneously in the same container. Food selection was examined by analysing the relationships over time between surviving termites and food consumption. The results indicated that R. grassei could be considered a generalist species, as it consumed the cork and wood of all oak species, as well as displaying a clear preference for soft wood (pine). Correlation analysis indicated that consumption was not dependent on wood density. Survival of R. grassei was influenced by the time of exposure to different oak species, but a high survival rate was maintained over time in the pine treatment (upper 70% in the three experiments). Given these results, it can be concluded that all the oak species are a suitable food source for R. grassei.

Since 2008, several outbreaks of yellow fever and dengue occurred in Abidjan, the economic capital of Côte d'Ivoire. A better knowledge of the biology of Aedes aegypti populations, the main vector of yellow fever and dengue viruses, is necessary to tailor vector control strategies implemented in the city. This study was designed to determine some biological parameters, occurring during the life cycle of two morphological phenotypes of Ae. aegypti larvae. Mosquitoes were sampled in a suburb of Abidjan (Treichville) using the WHO layer-traps technique. Biological parameters were studied in laboratory under standard conditions of temperature (27°C ± 2°C) and relative humidity (80% ± 10%). Our results indicated that the mean eggs laid by females from ‘brown larvae’ (BL) (85.95, 95% confidence interval (CI 95%) 78.87–93.02) was higher than those from ‘white larvae’ (WL) (64.40%, CI 95% 55.27–73.54). The gonotrophic cycle was 3 and 4 days in females from BL and WL, respectively. The overall yield of breeding mosquitoes from BL (63.88%, CI 95% 62.61–65.14) was higher compared with those of mosquitoes from WL (59.73%, CI 95% 58.35–61.12). The sex ratio (male/female) was 0.95 and 1.68 in Ae. aegypti populations from BL and WL, respectively. Females from BL lived slightly longer than those from WL (t = −2.332; P = 0.021). This study shows that Ae. Aegypti populations from BL and WL present different biological parameters during their life cycle. This could have an implication on their ability to transmit human disease viruses such as dengue and yellow fever. Further molecular studies are needed to determine genetic divergence between these Ae. aegypti populations.

In this study, we investigated resistance traits to the sugarcane borer Diatraea saccharalis Fab. (Lepidoptera: Crambidae) in the leaves and stalks of six sugarcane cultivars in a series of greenhouse and laboratory assays. Investigation of plant factors and infestation rates to better discriminate stalk damage by the sugarcane borer indicated that infestation of 7-month-old, single plants with 20 larvae at the third or fourth instar per plant was suitable to assess tunneling length. Three cultivars (i.e. SP803280, RB928064, and RB835486) had lower stalk damage (i.e. tunnel length) than cultivar SP891115, which exhibited relatively greater susceptibility to tunneling by the borer. The time required for the larvae to enter the sugarcane stalk was longer for cultivar SP803280, indicating resistance traits on the stalk surface, which correlated with lower stalk damage. Larvae feeding on SP813250 stalks had the lowest weight gain, indicating that this cultivar has resistance traits to larval development within its stalks. Cultivars RB867515 and SP891115 resulted in the highest mortality of early-stage larvae feeding on leaves, indicating the presence of resistance factors in their leaves. Multi-trait cluster and principal component analyses placed the cultivars into three and four clusters, respectively. The cultivars placed in different groups that exhibited resistance to leaf feeding, stalk entrance, and tunneling by the sugarcane borer could be used for crossings in sugarcane breeding programs with the goal of obtaining higher levels of resistance to D. saccharalis.

The purpose of this study was to investigate and compare root morpho-anatomical traits and physiological responses of susceptible (SP81–3250) and resistant (H. Kawandang) sugarcane genotypes exposed to the attack by nymphs of spittlebug Mahanarva fimbriolata (Stål) (Hemiptera: Cercopidae). Two experiments were conducted to compare the damage caused by spittlebug nymphs on fresh and dry biomass weight; lignin content in stalks; root anatomy; chlorophyll content; photosynthetic rate (A); carboxylation efficiency (A/Ci); stomatal conductance (gS) and transpiration rate (E) of these genotypes. SP81–3250 consistently obtained significantly higher damage scores than H. Kawandang in both experiments, confirming the previously observed level of resistance in each genotype. Attack by spittlebug nymphs had a much higher effect on both fresh and dry biomass weight, chlorophyll content, A, A/Ci, gs and E of SP81–3250, than that on H. Kawandang. Anatomical studies indicated the presence of aerenchyma tissue in the root cortex of SP81–3250, a feature which may facilitate penetration of the nymph's stylet into the vascular cylinder. In contrast, roots of H. Kawandang are characterized by having more dense and compact parenchyma cells. In addition, infested plants of this genotype contained an unidentified mucilaginous compound in the vascular cylinder of the roots. We conclude that resistance of H. Kawandang to spittlebug is related to the ability of this genotype to maintain normal chlorophyll content, as well as stomatal conductance and photosynthesis, thus, allowing for biomass accumulation under spittlebug attack, in contrast to SP81–3250. In addition, the presence of more compact and denser parenchymal cells, as well as that of an induced mucilaginous compound in the root's vascular cylinder, are likely to hinder host-feeding activity in nymphs, causing higher nymph mortality and therefore, reduced damage in plants of this genotype.