The aim of this study was to develop and optimize silverleaf bioassay, esterase analysis and PCR-based techniques to distinguish quickly and reliably biotype B of the whitefly, Bemisia tabaci (Gennadius), from Indian indigenous biotypes. Zucchini and squash readily develop silverleaf symptoms upon feeding by the B biotype, but they are not readily available in Indian markets. A local pumpkin variety ‘Big’ was, therefore, used in silverleaf assay, which developed symptoms similar to those on zucchini and squash and can be used reliably to detect B biotype. Analysis of non-specific esterases of B and the indigenous biotypes indicated both quantitative and qualitative differences in esterase patterns. Two high molecular weight bands were unique to B biotype and they occurred in abundance. These esterases were used to develop quick and field-based novel detection methods for differentiating B from the indigenous biotypes. Development of these simple and cost-effective protocols has wider application as they can be potentially used to identify other agricultural pests. Mitochondrial cytochrome oxidase I gene sequences and randomly amplified polymorphic DNA (RAPD) polymorphisms, generated using the primer OpB11, were also found useful for detecting B. tabaci biotypes. A B biotype-specific RAPD band of 800 bp was sequenced, which was used to a develop sequence characterized amplified region (SCAR) marker. The SCAR marker involved the development of B biotype-specific primers that amplified 550 bp PCR products only from B biotype genomic DNA. Silverleaf assay, esterases, RAPDs or a SCAR marker were used in combination to analyse whitefly samples collected from selected locations in India, and it was found that any of these techniques can be used singly or in combination to detect B biotype reliably. The B biotype was found in southern parts of India but not in the north in 2004–06.

The effect of pyrethroid resistance on the fitness of a laboratory strain of Anopheles funestus originating from southern Mozambique was evaluated by comparing the developmental and reproductive characteristics of a pyrethroid resistant strain with an insecticide susceptible strain. Fitness was evaluated in terms of fecundity, fertility, egg production, developmental time and life stage progression and survival. Of the eggs laid by females of the resistant strain, 81.5% hatched while only 66.9% were recorded in the susceptible strain. The time from egg hatch to adult emergence was longer for the resistant strain (15.9 days) than the susceptible strain (15.2 days). A significantly higher proportion of eggs from the resistant strain (61.6%) survived to adulthood compared with those of the susceptible strain (49%). Fecundity and larval and pupal survival did not differ significantly between strains. Of spermathecae dissected from females of the resistant strain, 56.8% were fertilized compared to 52.6% from the susceptible strain. The proportion of females that successfully produced eggs was 43.3% and 23.3% for the resistant and susceptible strains respectively. Complete failure of larval hatch was recorded in 28.6% of susceptible strain families compared to 7.7% of resistant families. Our results show that pyrethroid resistance in southern African An. funestus does not incur any loss of fitness under laboratory conditions. These results suggest that the removal of pyrethroid insecticide selection pressure may not lead to a regression of resistance alleles in pyrethroid resistant An. funestus populations in southern Africa.

The susceptibility of representative pyrethroid (cypermethrin, deltamethrin, lambdacyhalothrin, bifenthrin), organophosphate (chlorpyriphos, triazophos, profenophos) and new chemistry insecticides (spinosad, indoxacarb and emamectin) was investigated for 18 field populations of Plutella xylostella (Linnaeus) from three different zones in Pakistan. The LC50 (mg ml−1; 48 h) values of pyrethroids for various populations ranged from 0.19–1.88 for cypermethrin, 0.31–2.64 for deltamethrin, 0.08–1.16 for lambdacyhalothrin and 0.07–0.88 for bifenthrin. The LC50 (mg ml−1; 48 h) of organophosphates ranged from 0.52–5.67 for chlorpyriphos, 0.37–4.14 for triazophos and 0.03–2.65 for profenophos. The most probable reason for low toxicity of organophosphates and pyrethroids is the evolution of multiple resistance mechanisms; however, further studies are required to establish these mechanisms. When these same products were tested against a susceptible laboratory population (Lab-Pak), the new chemistry compounds were significantly more toxic than pyrethroids and organophosphates. The results are discussed in relation to integrated pest management and insecticide resistance management strategies for P. xylostella.

Ochlerotatus albifasciatus is a vector of western equine encephalomyelitis in Argentina and a nuisance mosquito affecting beef and dairy production. The objective of this study was to analyze whether environmental proxy data derived from 1 km resolution NOAA-AVHRR images could be useful as a rapid tool for locating areas with higher potential for Oc. albifasciatus activity at a regional scale. Training sites for mosquito abundance categories were 3.3×3.3 km polygons over sampling sites. Abundance was classified into two categories according to a proposed threshold for economic losses. Data of channels 1, 2, 4 and 5 were used to calculate five biophysical variables: normalized differences vegetation index (NDVI), land surface temperature, total precipitable water, dew point and vapour saturation deficit. A discriminant analysis correctly classified 100% of the areas predicted to be above or below the economic threshold of 2500 mosquitoes per night of capture, respectively. Components of the NDVI, the total precipitable water and the dew point temperature contributed most to the function value. The results suggest that environmental data derived from AVHRR-NOAA could be useful for rapidly identifying adequate areas for mosquito development or activity.

The population genetic structures of Culex pipiens Linnaeus were evaluated in China over a 2000 km transect that encompasses the two subspecies, C. p. pallens and C. p. quinquefasciatus. Four polymorphic allozyme loci were investigated in 1376 mosquitoes sampled from 20 populations across four provinces. These loci were not statistically dependent with no apparent heterozygote deficit or excess. On a regional scale (intra-province), a low (Fst=0.007–0.016) and significant genetic differentiation was found, with no clear geographical pattern. On a wider scale (inter-province), the genetic differentiation was higher (Fst=0.059), and an isolation by distance emerged. The results are compared with previous population genetic surveys of this mosquito species in different geographic areas over the world. The overall pattern suggests that Culex pipiens requires considerable distance (500–1000 km) to show isolation by distance, irrespective of the subspecies (C. p. pipiens, C. p. quinquefasciatus and C. p. pallens) or the geographic location.

Diamondback moth (DBM), Plutella xylostella, is known for causing damage to Brassica crops and developing resistance to chemical and biological pesticides; it has become one of the most difficult pests to manage in many regions around the world. The only way to reduce reliance on pesticides is to maximize the role of natural control agents for integrated pest management programs and be able to incorporate the mortality from control agents into pest control decision-making. More than 90 hymenopterous parasitoids are associated with DBM worldwide; among them, Diadegma semiclausum, is a major endoparasitoid of P. xylostella. To optimize parasitism of pests in pest control decision-making, it is necessary to develop rapid and simple methods for distinguishing parasitized from non-parasitized larvae in the field. Here we report on a number of diagnostic tools to identify parasitized larvae. One is based on differential melanization reactions in hemolymph due to immune suppression in parasitized larvae. The lack of coagulation reactions in hemolymph provides a simple initial test, where squashing a non-parasitized larva onto nitrocellulose membrane traps chlorophyll-containing gut content on the membrane leaving a green dot of clotted gut material. However, in immune-suppressed parasitized larvae, the gut content was washed away in absence of coagulation reactions and the membrane lacks a green dot. This tool alone or combined with others, allows quick detection of parasitized caterpillars in the field. We further showed that the antibody MAb 9A5 can be used to detect D. semiclausum parasitized larvae of DBM in Western blots.

Parasitoids employ different types of host-related volatile signals for foraging and host-location. Host-related volatile signals can be plant-based, originate from the herbivore host or produced from an interaction between herbivores and their plant host. In order to investigate potential sex- and species-related differences in the antennal response of parasitoids to different host-related volatiles, we compared the electroantennogram (EAG) responses of both sexes of the specialist parasitoid, Microplitis croceipes (Cresson), and the generalist, Cotesia marginiventris (Cresson), to varying doses of selected plant-based host-related volatiles: two green leaf volatiles (cis-3-hexenol and hexanal) and three inducible compounds (cis-3-hexenyl acetate, linalool, and (E,E)-α-farnesene). Mating had no significant effect on EAG response. Females of both species showed significantly greater EAG responses than conspecific males to green leaf volatiles, which are released immediately after initiation of herbivore feeding damage. In contrast, males showed greater responses than conspecific females to inducible compounds released much later after initial damage. Cotesia marginiventris females and males showed greater EAG responses than counterpart M. croceipes to the tested compounds at various doses, suggesting that the generalist parasitoid shows greater antennal sensitivity than the specialist to the tested host-plant volatiles. These results are discussed in relation to the possible roles of green leaf volatiles and inducible compounds in the ecology of female and male parasitoids.

Transgenerational maternal effects on performance (rm) after host transfer were evaluated in the generalist aphid Myzus persicae s.s., and in its subspecies specialized on tobacco, M. persicae nicotianae Blackman. We tested whether the performance of these taxa, when reared separately on optimal and suboptimal hosts (as sources of different maternal background) and then transferred to optimal hosts, experienced variations along four successive generations. Additionally, to compare the tolerance of both taxa to stress following host transfers, developmental instability (fluctuating asymmetry and body abnormalities) along the four generations was assessed. Taxon, rearing host, and generation affected the performance after host transfer. In the generalist, there was a significant improvement of rm along generations when transferred from suboptimal to optimal host and a significant decrease when transferred from optimal to optimal host; in the specialist, no increase or decrease occurred in any host transfer treatment. Transfer from suboptimal to optimal hosts caused higher losses of remaining replicates along generations than transfers from optimal to optimal hosts, and the specialist showed higher losses than the generalist. The only significant effect detected in comparisons involving fluctuating asymmetry values was that of taxon on length of siphunculi. Frequency of body abnormalities was not affected by treatments. Collectively, these results show a transgenerational weakening of maternal effects in the generalist but not in the specialist aphid, and suggest that rearing the latter in a suboptimal host causes not easily reversible changes that further give rise to constraints in performance.

The success or not of ladybirds as biological control agents is dependent on both their foraging behaviour and their individual survival rates. The former is a function of the habitats they utilise; the latter, a consequence of their reproductive strategy. Egg clustering was investigated in two ladybird species, Aphidecta obliterata, a conifer specialist, and Adalia bipunctata, an arboreal woodland generalist. The effect of oviposition substrate (filter papers vs. spruce needles) on clutch size and oviposition preference was also tested. Adalia bipunctata laid significantly more eggs than A. obliterata. The size of egg clusters laid by the two coccinellids varied between species and substrate types. Adalia bipunctata laid larger egg clusters than A. obliterata, with batches reaching a maximum size of 32 eggs on spruce and 41 eggs on paper, while those of A. obliterata contained a maximum of 5 eggs on spruce and 9 eggs on paper. Of the clusters laid by A. obliterata, 18.6% of those on paper and 21.4% of those on spruce contained only a single egg, whereas a minimum of two eggs per cluster were laid by A. bipunctata. Smaller clusters were laid on the spruce cuttings by both species when compared with those laid on the filter paper, but A. obliterata laid significantly more eggs on spruce than on the filter paper (77% vs. 23%), whilst A. bipunctata laid significantly more eggs on the filter paper (91%). It is suggested that coccinellid eggs are more likely to be washed off spruce needles than broad leaves and that, by laying smaller egg clusters on spruce, A. obliterata reduces this risk. Adalia bipunctata usually lays its eggs on the underside of broad leaved trees and thus does not face this risk and thus can lay larger egg clusters. No differences in cannibalism rates were found between the two species. These findings have implications for the use of ladybirds as biological control agents in spruce forests.

Geographic body size variation was analysed in males and females of 19 populations of the South American grasshopper Dichroplus vittatus Bruner spanning 20 degrees of latitude and 2700 m of altitude. Using mean and maximum body length of each sex and factors obtained from principal components analyses of six morphometric linear characters it was shown that D. vittatus followed the converse to Bergmann's rule latitudinally but not altitudinally where no significant trends were observed. For males, variability of body size increased with latitude but not with altitude. Both types of trends were significantly correlated with mean annual temperature and minimum annual temperature (positive correlations), and two estimators of seasonality, the coefficients of variation of mean annual temperature (negative) and mean annual precipitation (positive). Some allometric relationships also showed geographic variation. It is suggested that the observed decrease in size with latitude together with the increase in morphological variability is a consequence of a number of factors: the shortening of the growing season southwards; the increasing seasonality and climatic unpredictability; and the fact that the species exhibits protandry which contributes to smaller and more variable size in males and smaller but more constant body size in females.

The sweet potato whitefly, Bemisia tabaci, harbors Portiera aleyrodidarum, an obligatory symbiotic bacterium, as well as several secondary symbionts including Rickettsia, Hamiltonella, Wolbachia, Arsenophonus, Cardinium and Fritschea, the function of which is unknown. Bemisia tabaci is a species complex composed of numerous biotypes, which may differ from each other both genetically and biologically. Only the B and Q biotypes have been reported from Israel. Secondary symbiont infection frequencies of Israeli laboratory and field populations of B. tabaci from various host plants were determined by PCR, in order to test for correlation between bacterial composition to biotype and host plant. Hamiltonella was detected only in populations of the B biotype, while Wolbachia and Arsenophonus were found only in the Q biotype (33% and 87% infection, respectively). Rickettsia was abundant in both biotypes. Cardinium and Fritschea were not found in any of the populations. No differences in secondary symbionts were found among host plants within the B biotype; but within the Q biotype, all whiteflies collected from sage harboured both Rickettsia and Arsenophonus, an infection frequency which was significantly higher than those found in association with all other host plants. The association found between whitefly biotypes and secondary symbionts suggests a possible contribution of these bacteria to host characteristics such as insecticide resistance, host range, virus transmission and speciation.

Restriction fragment length polymorphism analysis of two segments of mitochondrial DNA (COI and 16S rRNA) was used to examine genetic variation in Sesamia nonagrioides (Lefèbvre) populations from the Mediterranean basin. Four populations were collected from central and southern Greece, and five from northern latitudes: Greece, Italy, France and Spain. No variation was observed in COI, while 16S rRNA segment proved highly polymorphic and 28 different haplotypes were found. Lower intra-population polymorphism was found in the northern populations than in southern ones. Although no significant isolation by distance was found, the UPGMA tree based on Nei's raw number of nucleotide differences separated the populations into two major groups, i.e. one with the northern (40.6°N–43.4°N) and the other with the southern populations (37.3°N–39.2°N). Analysis of molecular variance revealed that most of the variation was between the two major groups (ΦCT=0.559), and all pairwise comparisons between the northern and southern populations resulted in high and significant FST values (overall FST=0.604). The high FST values and the strong spatial genetic structure indicate that long-distance migration may be a rare event. The populations do not seem to have experienced a strong historical bottleneck. The occurrence of a few widespread haplotypes and the genetic similarity of the northern populations could be attributed to a historical expansion of certain haplotypes from the south towards to the northern borders of the species' distribution area.

The aim of this study was to determine which Arabidopsis thaliana (L.) genes had significantly altered expression following 2–36 h of infestation by the aphid Myzus persicae (Sulzer). Six biological replicates were performed for both control and treatment at each time point, allowing rigorous statistical analysis of any changes. Only two genes showed altered expression after 2 h (one up- and one down-regulated) while two were down-regulated and twenty three were up-regulated at 36 h. The transcript annotation allowed classification of the significantly altered genes into a number of classes, including those involved in cell wall modification, carbon metabolism and signalling. Additionally, a number of genes were implicated in oxidative stress and defence against other pathogens. Five genes could not currently be assigned any function. The changes in gene expression are discussed in relation to current models of plant-insect interactions.

The time during which prey remains are detectable in the gut of a predator is an important consideration in the interpretation of molecular gut-content data, because predators with longer detectability times may appear on the basis of unweighted data to be disproportionately important agents of prey population suppression. The rate of decay in detectability, typically expressed as the half-life, depends on many variables; one that has not been explicitly examined is the manner in which the predator processes prey items. The influence of differences in feeding mode and digestive physiology on the half-life of DNA for a single prey species, the Colorado potato beetle Leptinotarsa decemlineata (Say), is examined in two predators that differ dramatically in these attributes: the pink ladybeetle, Coleomegilla maculata (DeGeer), which feeds by chewing and then ingesting the macerated material into the gut for digestion; and the spined soldier bug, Podisus maculiventris (Say), which physically and enzymatically processes the prey extra-orally before ingestion and further digestion in the gut. In order to standardize the amount of DNA consumed per predator, a single L. decemlineata egg was used as the prey item; all predators were third instars. The PCR assay yields estimated prey DNA half-lives, for animals maintained under field temperatures, of 7.0 h in C. maculata and 50.9 h in P. maculiventris. The difference in the prey DNA half-lives from these two predators underscores the need to determine detectabilities from assemblages of predators differing in feeding mode and digestive physiology, in order to weight positives properly, and hence determine the predators' relative impacts on prey population suppression.

Two experiments were done to examine the predation of thrips, and the movement of Orius laevigatus Fieber and Neoseiulus cucumeris (Oudemans) in the presence and absence of two supplemental food sources, pollen and the fungus Trichoderma viride. The presence of pollen led to a 55% reduction in predation of the thrips by N. cucumeris and a 40% reduction in thrips predation by O. laevigatus, in experiments using single predators. The presence of fungus had no significant effect on thrips predation by either of the natural enemy species. Movement of the natural enemies was examined in a multiple predator experiment, and this showed that O. laevigatus was more likely to remain on the plant in the presence of thrips and when supplemental food, either pollen or fungus, was present. For N. cucumeris, there was no association between the presence of thrips and the mite, with the majority of the mites being found on the leaves where pollen was present. Although the single and multiple predator experiments were done at different times, the indications are that the predation rates of the N. cucumeris do not differ greatly between the two experiments, suggesting that there may be a potential interference effect between the mites, which is not present for O. laevigatus. The significance of these results for the use of supplemental food sources in biological control is discussed.

Similarities and differences in foraging behaviour between first and third instar Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) on Vigna radiata (L.) R. Wilczek (Fabaceae) (mung bean) were determined by frequent, repeated observation of insects on whole plants. The time apportioned to feeding, resting and searching by these two larval instars differed. Third instars were often found feeding on exposed areas of the plant and, if in the terminal parts, would leave these areas more frequently than first instars. Third instars fed at fewer sites and spent less time searching and resting, fed in longer bouts and spent 20% more time feeding than first instars. Although both instars tended to move upwards to the top of the plant, this applied to approximately half of the third instars observed compared to all of the first instars. First instars were found to have a higher relative growth rate than third instars. The results show that it cannot be assumed that first instars are just smaller versions of late instars.

Effects of the antiparasitic drug, ivermectin, on the dung beetles, Caccobius jessoensis Harold, 1867 and the rare species, Copris ochus Motschulsky, 1860 and Copris acutidens Motschulsky, 1860 were studied in laboratory and field experiments in Hokkaido, Japan. Ivermectin was detected in dung from 1 to 21 or 28 days following treatment, with a peak on the first day after treatment in two pour-on administrations (500 μg kg−1), although there were considerable differences between the two peaks. In C. jessoensis, brood balls constructed by the female were not reduced in the dung of treated cattle except for seven days after treatment in experiment 2. Also, there was no significant difference in the mean weight of brood balls between dung from treated and control cattle. However, the emergence rates were significantly reduced in dung 1–3 days after treatment. In the field study, brood balls constructed by C. jessoensis were more abundant in dung from treated cattle in experiment 1, but adult emergence was significantly reduced at one and seven days after treatments. Adult mortality of C. ochus Motschulsky at 90 days after the beginning of rearing was 11.1% in dung from control cattle with 22 brood balls constructed, whereas it was 84% in dung from treated cattle with no brood balls and/or ovipositioning. Also, in C. acutidens Motschulsky, adult mortality at 90 days after the beginning of rearing was 3.6% in dung from control cattle with 13 brood balls constructed, whereas it was 94.1% in dung from treated cattle with no brood balls or ovipositioning. The environmental risk in the use of ivermectin during breeding period of dung beetles in pasture is discussed.

Nilaparvata lugens Stål, the Brown Planthopper (BPH), is a major pest in rice. The lycosid Pardosa pseudoannulata (Bösenberg & Strand) and the linyphiid Atypena formosana (Oi) are the early dominant predators in irrigated rice. Later, predatory bugs, including the mirid Cyrtorhinus lividipennis Reuter, become dominant. In unsprayed rice, BPH numbers normally remain low. While P. pseudoannulata is known to be a key natural enemy of BPH, the contribution to BPH reduction by the smaller A. formosana is less well known. Due to the size difference, A. formosana may also be an intraguild prey of P. pseudoannulata. To investigate predation on BPH by the two spider species alone, together and in combination with C. lividipennis, two cage experiments were conducted, the first with adult spiders, the second with immature/unmated spiders. In the latter, spiders were introduced with a delay to allow a better establishment of C. lividipennis. In both experiments, BPH numbers were lowest in treatments with more than one predator species present. Intermediate BPH reduction was obtained with P. pseudoannulata followed by A. formosana and C. lividipennis. While P. pseudoannulata can prey upon large nymphs and adult BPH, A. formosana favour early instar BPH, leading to a more even control across instars in treatments with both spider species. Pardosa pseudoannulata numbers increased more with A. formosana or A. formosana and C. lividipennis present. In contrast, P. pseudoannulata reduced A. formosana and C. lividipennis numbers. The presence of C. lividipennis apparently lessened intraguild predation on A. formosana and may be an intraguild prey of A. formosana. Competition for prey may have added to the antagonistic interactions found. From a biological control perspective, the presence of both spiders in early rice is an advantage for the biological control of BPH in rice. In early rice, results show that A. formosana is an important predator of BPH and, like C. lividipennis, can complement control by P. pseudoannulata and serve as intraguild prey for the latter species.

Recent studies show that Vetiver grass, (Vetiveria zizanioides (L.) Nash), may have potential as a dead-end trap crop in an overall habitat management strategy for the spotted stem borer, Chilo partellus (Swinhoe) (Lepidoptera: Crambidae). Vetiver grass is highly preferred for oviposition, in spite of the fact that larval survival is extremely low on this grass. The oviposition behaviour of female Chilo partellus moths was investigated by determining the amount and size of egg batches allocated to maize and Vetiver plants and studying the effect of rearing conditions and oviposition experience on host plant selection. Two-choice preference tests were used to examine the effect of experience of maize (a suitable host plant) and Vetiver plants on the oviposition choice of C. partellus. For both field-collected and laboratory-reared moths, no significant differences were found in the preference distributions between the experienced groups. It is concluded that females do not learn, i.e. that they do not change their preference for Vetiver grass after having experienced oviposition on either maize or this grass, which supports the idea that trap cropping could have potential as a control method for C. partellus. Differences observed between field-collected and laboratory-reared moths in the amount and size of egg batches laid on maize and Vetiver grass indicate that data obtained from experiments with laboratory-reared insects should be treated with caution.

As consequence of the concern about the biosafety of genetically modified plants, biological and ecological studies are considered crucial for environmental risk assessment. Laboratory experiments were carried out in order to evaluate the transfer of the Cry1Ac Bt-toxin from a transgenic Bt-oilseed rape to a non-target pest, Myzus persicae Sulzer. Cry1Ac protein levels in plants and aphids were determined using a double sandwich enzyme-linked immunosorbent assay. Phloem sap from (Bt+) and (Bt−) oilseed rape plants was collected from leaves using a standard method of extraction in an EDTA buffer. Bt-toxin was present in phloem sap, with a mean concentration of 2.7±1.46 ppb, corresponding to a 24-fold lower level than in oilseed rape leaves. Toxin was also detected in aphid samples, with a mean concentration in the positive samples of 2.0±0.8 ppb. The evidence that Bt-toxin remains in herbivores, in this case an aphid, could be useful to clarify functional aspects linked to possible consequences of Bt-crops on food chains involving herbivore–natural enemy trophic systems. Further studies are needed in order to improve the knowledge on the functional aspects linked to the transfer of the Cry1Ac Bt-toxin from GM-oilseed rape to aphids and their possible consequence.