Chironomids are excellent biological indicators for the health of aquatic ecosystems, but their use at finer taxonomic levels is hindered by morphological similarity of species at each life stage. Molecular markers have the potential to overcome these problems by facilitating species identification particularly in large-scale surveys. In this study, the potential of the polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) approach was tested to rapidly distinguish among chironomids within a geographic area, by considering chironomid species from Melbourne, Australia. By comparing molecular markers with diagnostic morphological traits, RFLP profiles of the cytochrome oxidase I (COI) region were identified that were specific to genera and some common species. These profiles were used to develop an RFLP–based key, which was validated by testing the markers on samples from several wetlands and streams. As well as allowing for rapid identification of species that are difficult to separate on morphological grounds, this approach also has the potential to resolve current taxonomic ambiguities.

The subterranean termites in the genus Reticulitermes have a complex and plastic life cycle, which has been the subject of a number of publications over the past century. Given the inherent difficulties in studying such cryptic, eusocial organisms it is not perhaps surprising that the literature on their biology has failed to reach a consensus. An overview of the literature is given, which is followed by a discussion of the various theories on the life cycle of Reticulitermes spp. A substantial proportion of the review focuses on the French literature, which constitutes the majority of the primary sources and can be difficult to access. There are many discrepancies in the literature in terms of the number of instars, the definition of workers and the question of whether they should be termed pseudergates or, potentially, an additional terminology used to differentiate between pseudergates and the true workers seen in the higher termites (Isoptera: Termitidae). It remains very difficult to compare publications as there is little conformity; a problem that is aggravated by a general absence of drawings of the relevant instars. Further work on the biology of Reticulitermes is clearly required. There is also a need for researchers to agree on a standard terminology for this genus. A glossary is provided for the various synonyms and definitions.

Bemisia tabaci (Gennadius) populations, collected from cassava and other plants in major cassava-cultivation areas of Sub-saharan Africa and from elsewhere around the world, were studied to determine their biotype status and genetic variation. Random amplified polymorphic DNA–polymerase chain reaction (RAPD–PCR) markers were used to examine the genetic structure of the populations. The dendogram obtained using the neighbour joining method (NJ) split the cassava-associated populations from the non-cassava types with a 100% bootstrap probability. Analysis of molecular variance (AMOVA) of the RAPD fragments revealed that 63.2% of the total variation was attributable to differences among populations, while the differences among groups (host) and within populations accounted for 27.1 and 9.8% respectively. Analysis of the internally transcribed spacer region I (ITS 1) of the ribosomal DNA confirmed that the cassava populations of B. tabaci populations were distinct from non-cassava populations. Experiments to establish whitefly populations on various host plants revealed that cassava-associated populations were restricted to cassava only, whereas B. tabaci from other hosts were polyphagous but did not colonize cassava. Hence, populations of B. tabaci from cassava in Africa represent a distinct group.

Global warming may affect the future pattern of many arthropod-borne diseases, yet the relationship between temperature and development has been poorly described for many key vectors. Here the development of the aquatic stages of Africa's principal malaria vector, Anopheles gambiae s.s. Giles, is described at different temperatures. Development time from egg to adult was measured under laboratory conditions at constant temperatures between 10 and 40°C. Rate of development from one immature stage to the next increased at higher temperatures to a peak around 28°C and then declined. Adult development rate was greatest between 28 and 32°C, although adult emergence was highest between 22 and 26°C. No adults emerged below 18°C or above 34°C. Non-linear models were used to describe the relationship between developmental rate and temperature, which could be used for developing process-based models of malaria transmission. The utility of these findings is demonstrated by showing that a map where the climate is suitable for the development of aquatic stages of A. gambiae s.s. corresponded closely with the best map of malaria risk currently available for Africa.

Cacopsylla pyricola (Förster) is one of the most important pests of pear in North America, where several native predators have been considered for integrated pest management (IPM) programmes. Two molecular markers of 271 and 188 bp were developed from C. pyricola cytochrome oxidase I (COI) fragments, in order to study the detection of this species in the gut of arthropod predators. Primer sensitivity and the detection period for pear psylla remains in the guts of Anthocoris tomentosus Pericart were determined. The sensitivity threshold was defined at 10-5 dilution of a C. pyricola fifth-instar nymph in all samples. Predator adults were evaluated immediately after ingestion of one to five C. pyricola nymphs (t = 0) and after 2, 4, 6, 8, 16, 24 and 32 h. Detection of the presence of C. pyricola DNA always lasted longer using the shorter fragment and was observed after 32 h of digestion using both markers. The primers amplifying the 188 bp fragment amplified all four psyllid species tested, whereas the primers designed to amplify the 271 bp fragment did so exclusively for C. pyricola and its close relative, Cacopsylla pyri (Linnaeus). Both primers failed to amplify DNA from representative species of the Coccinellidae, Chrysopidae, Hemerobiidae, Anthocoridae, Miridae, Salticidae, Aphididae, Tetranychidae and the Tortricidae, suggesting their suitability for general trophic studies.

The possibility to cross-species amplify microsatellites in fruit flies of the genus Ceratitis was tested with the polymerase chain reaction (PCR) by analysing 23 Ceratitis capitata (Wiedemann) microsatellite markers on the genomic DNA of three other economically important, congeneric species: C. rosa (Karsch), C. fasciventris (Bezzi) and C. cosyra (Walker). Twenty-two primer pairs produced amplification products in at least one of the three species tested. The majority of the products were similar, if not identical in size to those expected in C. capitata. The structures of the repeat motifs and their flanking sequences were examined for a total of 79 alleles from the three species. Sequence analysis revealed the same repeat type as the homologous C. capitata microsatellites in the majority of the loci, suggesting their utility for population analysis across the species range. A total of seven loci were differentially present/absent in C. capitata, C. rosa, C. fasciventris and C. cosyra, suggesting that it may be possible to differentiate these four species using a simple sequence repeat-based PCR assay. It is proposed that medfly-based microsatellite markers could be utilized in the identification and tracing of the geographical origins of colonist pest populations of the four tested species and in the assessment of their risk and invasive potentials; thereby assisting regulatory authorities in implementing quarantine restrictions and other pest control measures.

Bruchid pests such as Callosobruchus chinensis (Linnaeus) endanger stored legume seeds throughout the tropical belt. The chemical composition of the headspace volatiles from healthy and fourth instar larvae-infested cowpea seeds were identified, characterized, and compared using gas chromatography–mass spectrometry. Y-tube olfactometer bioassays were performed to evaluate the effect of these chemicals on the orientation of conspecific adult females. Analysis of volatiles released from healthy and infested seeds revealed qualitative differences for three out of the 17 compounds identified. Dimethyl disulphide, isobutenyl methyl ketone and methyl trisulphide were found only in the blend emitted from infested but not from healthy seeds. Quantitative differences were apparent for tridecane which was released in larger amounts from infested seeds. While volatiles collected from healthy seeds were attractive to female bruchids, volatiles collected from infested seeds were repellent. To test the hypothesis that the qualitative differences in the chemical composition found may be due to insect-derived components, the volatiles from frass and fourth instar larvae combined were analysed. These volatiles contained both of the sulphides emitted from infested seed but not from healthy seeds. Although a limited induction of volatiles from cowpea seeds cannot be excluded, it is postulated that behavioural differences of the female weevils are largely due to insect-derived semiochemicals. The potential use of such semiochemicals as part of an integrated pest management strategy is discussed.

Only pyrethroid insecticides have so far been recommended for the treatment of mosquito nets for malaria control. Increasing resistance of malaria vectors to pyrethroids threatens to reduce the potency of this important method of vector control. Among the strategies proposed for resistance management is to use a pyrethroid and a non-pyrethroid insecticide in combination on the same mosquito net, either separately or as a mixture. Mixtures are particularly promising if there is potentiation between the two insecticides as this would make it possible to lower the dosage of each, as has been demonstrated under laboratory conditions for a mixture of bifenthrin (pyrethroid) and carbosulfan (carbamate). The effect of these types of treatment were compared in experimental huts on wild populations of Anopheles gambiae Giles and the nuisance mosquito Culex quinquefasciatus Say, both of which are multi-resistant. Four treatments were evaluated in experimental huts over six months: the recommended dosage of 50 mg m−2 bifenthrin, 300 mg m−2 carbosulfan, a mosaic of 300 mg m−2 carbosulfan on the ceiling and 50 mg m−2 bifenthrin on the sides, and a mixture of 6.25 mg m−2 carbosulfan and 25 mg m−2 bifenthrin. The mixture and mosaic treatments did not differ significantly in effectiveness from carbosulfan and bifenthrin alone against anophelines in terms of deterrency, induced exophily, blood feeding inhibition and overall mortality, but were more effective than in earlier tests with deltamethrin. These results are considered encouraging, as the combination of different classes of insecticides might be a potential tool for resistance management. The mixture might have an advantage in terms of lower cost and toxicity.

Delta and wing traps baited with synthetic female sex pheromone of Leucinodes orbonalis Guenée were found to catch and retain ten times more moths than either Spodoptera or uni-trap designs. Locally-produced water and funnel traps were as effective as delta traps, although ‘windows’ cut in the side panels of delta traps significantly increased trap catch from 0.4 to 2.3 moths per trap per night. Trap catch was found to be proportional to the radius of sticky disc traps in the range 5–20 cm radius, discs with a 2.5 cm radius caught no moths. Wing traps placed at crop height caught significantly more moths than traps placed 0.5 m above or below the crop canopy. Replicated integrated pest management (IPM) trials (3 × 0.5 ha per treatment) were conducted in farmers fields with young and mature eggplant crops. Farmers applied insecticides at least three times a week in all check and IPM plots. In addition pheromone traps were placed out at a density of 100 per ha and infested shoots removed weekly in the 0.5 ha IPM plots. Pheromone trap catches were reduced significantly from 2.0 to 0.4 moths per trap per night respectively in check and IPM plots in a young crop and 1.1 to 0.3 moths per trap per night in check and IPM plots respectively in a mature crop. Fruit damage was significantly reduced from an average of 41.8% and 51.2% in check plots of young and mature crops respectively to 22% and 26.4 respectively in the associated IPM plots. Significant differences in pheromone trap catches and fruit damage were attained four and two weeks respectively after IPM treatments began in the mature crop whereas in the immature crop significant differences were not observed for the first eight to nine weeks respectively. The relative impact of removing infested shoots and mass trapping on L. orbonalis larval populations was not established in these trials but in both cases there was an estimated increase of approximately 50% in marketable fruit obtained by the combination of control techniques compared to insecticide treatment alone.

A single strain of Wolbachia (α-proteobacteria, Rickettsiales) was found in widespread geographical populations of each of two Phlebotomus species, within which there was no indication of ‘infectious speciation’. The two strains were identified by sequencing a fragment of wsp (a major surface protein gene), amplified by polymerase chain reaction from DNA extracted from the body parts of individual sandflies. Infection rates were high in the males and females of both sandflies, but they were lower for the B-group wPrn strain of Wolbachia in Phlebotomus perniciosus Newstead (60.3% overall) than for the A-group wPap strain in P. papatasi (Scopoli) (81.7%). Infections were frequent in the thorax, where Leishmania develops infective forms, as well as in the abdomen, where Wolbachia must infect the reproductive tissues to ensure its vertical transmission. These findings were related to knowledge of the population biology of Wolbachia in other insects, leading to the conclusion that this endosymbiont could be useful for driving transgenes through wild populations of both sandflies. This will require characterizing the cytoplasmic incompatibility phenotypes of Wolbachia–sandfly combinations, as well as estimating for them the incidence of paternal transmission and the fidelity of maternal transmission. Paternal transmission is one explanation for finding a single Wolbachia strain associated with all mitochondrial haplotypes and lineages of each sandfly species. However, this distribution pattern could also result from multiple horizontal transmissions or the failure of wsp to provide strain markers.

Methodology for detailed dissections of the phylogenetically most informative structures from adult aleocharine staphylinid beetles is provided. These methods include specimen preparation, relaxation, clearing, bleaching, dehydration and full dissection. A detailed summary of mouthpart and male and female genitalic dissections is provided. Recently described character systems, including those of the mouthparts and the thoracic regions, have been successfully used in phylogenetic studies of aleocharine staphylinids. This paper provides the first thorough description of the techniques necessary to reveal these character systems.

A study was undertaken to identify the major larval habitats of the Anopheles gambiae (Giles) complex in rural Gambia. Mosquito larvae and pupae were sampled along transects and in specific habitats in the central region of the country during the rainy seasons of 1996 and 1997. The sampling showed that the major breeding sites were located on the flooded alluvial soils bordering the river. The largest numbers of larvae were found during September, one month after the peak rains. Polymerase chain reaction analysis of specimens showed that Anopheles melas (Theobald) was the dominant species in the flooded areas (81.5%), followed by A. gambiae sensu stricto (Giles) (18.0%) and A.arabiensis (Patton) (0.5%). By sampling in specific habitats it was evident that A.arabiensis was mainly breeding in rain-fed rice fields along the edge of the alluvial soils. Anophelesmelas and A.gambiae s.s. often coexisted but whereas A. melas were found in water with a salinity of up to 72% sea water (25.2 g NaCl l−1), A.gambiae s.s. only occurred in water with up to 30% sea water (10.5 g NaCl l−1). Anophelesmelas larvae were found in association with plant communities dominated by sedges and grasses (Eleocharis sp., Paspalum sp., Sporobolus sp.) and sea-purslane Sesuviumportulacastrum (L.) and the presence of cattle hoof prints, whereas A.gambiae s.s. larvae mainly occurred in association with Paspalum sp. and Eleocharis sp. The study showed that even during the peak rainy season, breeding of the A.gambiae complex is almost entirely restricted to the extensive alluvial areas along the river.

In 1984–1985 insecticide-treated targets were deployed in the 600-km2 Rifa Triangle, Zambezi Valley, Zimbabwe. Trap catches of Glossina pallidipes Austen were modelled using a function combining logistic growth with diffusive movement. A simulation routine was linked to a non-linear least-squares optimization programme and fits optimized with respect to population carrying capacities, rates of growth and movement, and to levels of imposed mortality. In March–September 1984, the overall additional mortality was 2% per day of adult female G. pallidipes, increasing thereafter to 8% per day, due to the deployment of more targets, the onset of the hot, dry season and the ground-spraying of the adjoining Zambezi escarpment with DDT. For G. m. morsitans Westwood the corresponding estimates were 1 and 2% per day. For both species, the deployment of four targets km−2 in a closed population will ensure eradication. For G. m. morsitans a halving of target efficacy would reduce the killing rate to the point where eradication would be unlikely. Estimated daily displacements were c. 200 m for G. m. morsitans and 660 m for G. pallidipes. The lower rate for G. m. morsitans means that, while targets kill this species less effectively, re-invasion of cleared areas is slower. Targets do not markedly affect robust populations outside the deployment area. The Zambian tsetse population adjacent to the Rifa Triangle declined markedly during the experiment, however, suggesting that it is largely maintained by immigration. The methods developed here will be applied to data from other campaigns with the aim of improving the efficiency of tsetse control programmes.

Grape phylloxera, Daktulosphaira vitifoliae Fitch, is an important pest of grapevines (Vitis vinifera L.) (Vitaceae). Using microsatellite DNA markers it was demonstrated strong associations can exist between D. vitifoliae asexual lineages and vine host type within a vineyard. Also, in excised root bioassays, D. vitifoliae collected from three regions where different genotypic classes predominated showed host-specific differences in life table parameters of reproductive rate and intrinsic rate of increase. Lastly, comparisons of mitochondrial DNA (cytochrome oxidase I) sequences revealed that D. vitifoliae in Australia have paraphyletic origins and fall into two clades partially related to vine host usage. These findings indicate introduction of separate lineages of D. vitifoliae which have close host associations and as such, have important implications for management of this pest in Australia.

Intensive chemical treatments have led to the development of a number of insecticide resistance mechanisms in the peach–potato aphid Myzus persicae (Sulzer). Some of these mechanisms are known to be associated with negative pleiotropic effects (resistance costs). Molecular and biochemical methods were used to determine the genotypes or phenotypes associated with four insecticide resistance mechanisms in single aphids from sexually-reproducing populations in southern France. The mechanisms considered were E4 and FE4 carboxylesterase overproduction, modified acetycholinesterase, and kdr and rdl resistance-associated mutations. A new method for determining individual kdr genotypes is presented. Almost all resistant individuals overproduced FE4 carboxylesterase, whereas modified acetylcholinesterase was rare. Both the kdr and rdl resistance mutations were present at high frequencies in French sexually-reproducing populations. The frequencies of insecticide resistance genes were compared before and after sexual reproduction in one peach orchard at Avignon to evaluate the potential impact of selection on the persistence of resistance alleles in the over-wintering phase. The frequencies of the kdr and rdl mutations varied significantly between autumn and spring sampling periods. The frequency of the kdr mutation increased, probably due to pyrethroid treatments at the end of the winter. Conversely, the frequency of the rdl mutation decreased significantly during winter, probably because of a fitness cost associated with this mutation.

The autumn return migration of the beet armyworm Spodoptera exigua Hübner and other insects in northern China was observed with radar and with a simultaneously-operated searchlight trap and ground light-trap at a site in Langfang, near Beijing. The searchlight trap was found to be an efficient tool for trapping migrants and, operated alongside a ground light-trap, could distinguish migrant from locally-flying species. It was confirmed that S. exigua and some other species were high-altitude nocturnal windborne migrants during September and October in northern China. Maximum density of moths typically occurred below 500 m, and strong layering was often observed at about 200 m above ground level in airflows that would carry the moths towards the south. Descent of S. exigua in the vicinity of the radar site in late September was often associated with rain.

Laboratory experiments examined interspecific interactions between larvae of three coccinellid species, Curinus coeruleus Mulsant (Chilocorinae), Harmonia axyridis Pallas and Olla v-nigrum (Mulsant) (Coccinellinae), and between these and larvae of the green lacewing, Chrysoperla rufilabris (Burmeister). Larvae of C. coeruleus, although defended on their dorsal surface with long spines, had the smallest mandibles, were the slowest-moving, and the least successful in interspecific larval combat. The long spines of third instar C. coeruleusappeared to reduce their palatability as food to H. axyridis and O. v-nigrum larvae in choice tests with dead larvae, but were not an effective defence against these species in Petri dish arenas. Larvae of O. v-nigrum had a smooth dorsal surface, were intermediate in terms of mandible size, but were the fastest moving, a trait that benefited their survival in intraguild combat. Larvae of H. axyridis were intermediate with respect to dorsal spines and speed of movement, but had the largest mandibles. This species was the most effective intraguild combatant among the coccinellids and the only one to successfully compete against C. rufilabris larvae of similar age. The speed, manoeuverability and long mandibles of C. rufilabris enabled them to impale coccinellid larvae at a relatively safe distance. The spines of C. coeruleus larvae impeded laterally oriented attacks by C. rufilabris, but did not provide sustained protection from repeated attacks. Success in these interactions appeared largely a function of offensive weaponry (mandible size and morphology) and speed of movement, although the role of dorsal spines as defensive structures was not ruled out. Rates of larval cannibalism were highest for C. rufilabris and largely mirrored the level of aggression observed in interspecific combat for each species.

Variation in the effectiveness of biocontrol agents on the weed Parthenium hysterophorus L. was evaluated at two properties (Mount Panorama and Plain Creek) in Queensland, Australia for four years (1996–2000) using a pesticide exclusion experiment. At Mount Panorama, higher levels of defoliation by the leaf-feeding beetle Zygogramma bicolorata Pallister and galling by the moth Epiblema strenuana Walker in 1996–97 coincided with an above average summer rainfall, but in the following three years with below average summer rainfall the defoliation and galling levels were significantly lower. Biocontrol had significant negative impact on the weed only in 1996–97 with no major impact in the following three years. At Plain Creek, galling by E. strenuana was evident in all the four years, but varied significantly between years due to non-synchrony between P. hysterophorus germination and E. strenuana emergence. At Plain Creek biocontrol had limited impact on the weed in 1996–97 and 1997–98, with no significant impact in the following two years. Over the 4-year period, defoliation and galling resulted in 70% reduction in the soil seed bank at Mount Panorama, but the reduction in the soil seed bank at Plain Creek due to galling was not significant. Effectiveness of Z. bicolorata and E. strenuana was dependent on weather conditions and as a result had only limited impact on the weed in three out of four years.

Laboratory bioassays and field trials were used to characterize resistance to three aphid species (Myzus persicae (Sulzer), Acyrthosiphon kondoi Shinji, Aphis craccivora (Koch) in two aphid-resistant varieties (Kalya, Tanjil) and one susceptible variety (Tallerack) of Lupinus angustifolius L., and in one resistant variety (Teo) and one susceptible variety (Wodjil) of L. luteus L. Host selection tests in the glasshouse showed that alates of all three species preferred L. luteus to L. angustifolius, but provided no evidence that alates selected susceptible varieties over resistant. These results were supported by a field trial, which showed no difference in the number of colonizing A. kondoi alates collected from the resistant and susceptible lines of each lupin species, but there were significantly more late-instar nymphs and apterous adults on the susceptible lines. In laboratory host suitability experiments, there was much greater suppression of aphid growth and survival on Teo than on Kalya and Tanjil. In field trials, the numbers of aphids were generally lower on resistant compared to susceptible lines of both lupin species with one notable exception: M. persicae numbers were not lower on the resistant variety Tanjil compared to the susceptible variety Tallerack (L. angustifolius). These results suggest that the resistance mechanisms in both lupin species do not affect the selection of hosts by colonizing aphids, but rather are affecting the growth, survival and possibly reproduction of aphids after settling.

The separation of the closely related predatory species Macrolophus melanotoma Costa (= M. caliginosus Wagner) and Macrolophus pygmaeus (Rambur) based exclusively on the different colour pattern of the first antennal segment (white central band in M. melanotoma and entirely black in M. pygmaeus) is rather precarious and their taxonomic status is still in doubt. In the present study their taxonomic status was evaluated by DNA confirmatory analysis and hybridization experiments between M. pygmaeus and a Macrolophus taxon, resembling M. melanotoma, with a first antennal segment entirely black or with a white central band collected from Dittrichia viscosa (L.) W. Greuter (Asteraceae) in southern Greece. Adult females from Dittrichia plants hybridized with males of M. pygmaeus and vice versa did not produce viable eggs. The Macrolophus species from Dittrichia irrespective of the first antennal segment coloration differed from M. pygmaeusin digestive patterns generated by AseI, XbaI, and MseI on 16S rRNA and in RAPD profiles produced by the primers OPA-18 and OPA-20. These results demonstrate that on Dittrichia plants there is a distinct dimorphic taxon, M. melanotoma, as it is the only species of the genus Macrolophus bearing a first antennal segment with a central white band. Given the limitation of the coloration pattern, the mtDNA genetic markers are the appropriate method for the identification of M. melanotomaand M. pygmaeus.