Control of the malaria vector An. gambiae is still largely obtained through chemical intervention using pyrethroids, such as permethrin. However, strains of An. gambiae that are resistant to the toxic effects of pyrethroids have become widespread in several endemic areas over the last decade. The objective of this study was to assess differences in five life-history traits (larval developmental time and the body weight, fecundity, hatch rate, and longevity of adult females) and energy metabolism between a strain of An. gambiae that is resistant to permethrin (RSP), due to knockdown resistance and enhanced metabolic detoxification, and a permethrin susceptible strain reared under laboratory conditions. We also quantified the expression levels of five antioxidant enzyme genes: GSTe3, CAT, GPXH1, SOD1, and SOD2. We found that the RSP strain had a longer developmental time than the susceptible strain. Additionally, RSP adult females had higher wet body weight and increased water and glycogen levels. Compared to permethrin susceptible females, RSP females displayed reduced metabolic rate and mitochondrial coupling efficiency and higher mitochondrial ROS production. Furthermore, despite higher levels of GSTe3 and CAT transcripts, RSP females had a shorter adult life span than susceptible females. Collectively, these results suggest that permethrin resistance alleles might affect energy metabolism, oxidative stress, and adult survival of An. gambiae. However, because the strains used in this study differ in their genetic backgrounds, the results need to be interpreted with caution and replicated in other strains to have significant implications for malaria transmission and vector control.

Stink bugs (Heteroptera: Pentatomidae), including Nezara viridula (L.), Euschistus servus (Say), and Chinavia hilaris (Say), are economic pests in farmscapes where they move within and between closely associated crop and non-crop habitats. Thus, field edges in these farmscapes include not only crop-to-crop interfaces but also those edges adjoining non-crop habitats. We examined the influence of field edges on colonization of stink bugs in southeastern USA farmscapes composed of typical combinations of corn, peanut, and cotton. For E. servus and N. viridula, egg-to-adult development and presence of both sexes on all crops indicated that the crops served as reproductive plants. Adult C. hilaris were rarely found on corn and on crops associated with it, and they were present mainly in cotton in peanut–cotton farmscapes. Mature crop height was significantly higher for corn than for cotton and significantly higher for cotton over peanut, and an edge effect in dispersal of stink bugs into a crop was detected up to 4.6, 8.2, and 14.6 m from the crop-to-crop interface in corn, cotton, and peanut, respectively. These results suggest that stink bug dispersal into a crop decreases as crop height increases. The first stink bug-infested crop at the crop-to-crop interface was the most significant contributor of colonizing stink bugs to an adjacent crop. An edge effect in dispersal of stink bug adults was detected in corn next to non-woodlands and woodlands and in cotton adjacent to woodlands. Edge effects were never detected in side edges of peanut. Overall, our results indicate that both plant height and host plant suitability can influence edge-mediated dispersal of stink bugs at field edges.

Rhynchophorus ferrugineus (Coleoptera, Curculionidae) is the most destructive pest of palm trees worldwide containing it invasive areas, such as the southern part of China. It is always emphasized to develop integrated pest management based on biological agents, but their success is not very exciting. Presently, the immune defenses of this pest against biological agents attract scarce attention. It is still unclear whether immune priming also generally occurs in insect pests and in response to different pathogens. Our results indicated that previous challenge of bacteria pathogen enhanced the magnitude of phenoloxidase activity and antibacterial activity in R. ferrugineus larvae against the secondary infection. Furthermore, trans-generational immune priming was also determined in this pest, and only challenged R. ferrugineus mothers transferred the immune protection to their offspring which suggested males and females of this pest might have evolved different strategies on the investment of delivering immune protection to their offspring. Importantly, our data provide the evidence to suggest that different kinds of biological control agents might be used alternatively or in combination to fight against R. ferrugineus because of the existence of immune priming with low species-specific level. On the other hand, for this invasive pest, the immune priming may also facilitate its adaptation and dispersal in the new regions.

Reproductive interference is one of the major factors mediating species exclusion among insects. The cryptic species Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) of the whitefly Bemisia tabaci complex have invaded many parts of the world and often exhibit niche overlap and reproductive interference. However, contrasting patterns of competitive displacement between the two invaders have been observed between regions such as those in USA and China. Understanding the roles of reproductive interference in competitive interactions between populations of the two species in different regions will help unravel other factors related to their invasion. We integrated laboratory population experiments, behavioural observations and simulation modelling to investigate the role of reproductive interference on species exclusion between MEAM1 and MED in China. In mixed cohorts of the two species MEAM1 always excluded MED in a few generations when the initial proportion of MEAM1 was ⩾0.25. Even when the initial proportion of MEAM1 was only 0.10, however, MEAM1 still had a higher probability of excluding MED than that for MED to exclude MEAM1. Importantly, we show that as MEAM1 increased in relative abundance, MED populations became increasingly male-biased. Detailed behavioural observations confirmed that MEAM1 showed a stronger reproductive interference than MED, leading to reduced frequency of copulation and female progeny production in MED. Using simulation modelling, we linked our behavioural observations with exclusion experiments to show that interspecific asymmetric reproductive interference predicts the rate of species exclusion of MED by MEAM1. These findings not only reveal the importance of reproductive interference in the competitive interactions between the two invasive whiteflies as well as the detailed behavioural mechanisms, but also provide a valuable framework against which the effects of other factors mediating species exclusion can be explored.

Individual wild-caught sandflies from Iran were examined for infections of Wolbachia pipientis by targeting the major surface protein gene wsp of this intracellular α-proteobacterium. In total, 638 male and female sandflies were screened, of which 241 were found to be positive for one of three wsp haplotypes. Regardless of geographical origins and habitats, Phlebotomus (Phlebotomus) papatasi and other sandfly species were found to be infected with one common, widespread strain of A-group W. pipientis (Turk 54, GenBank accession EU780683; AY288297). In addition, a new A-group haplotype (Turk07, GenBank accession KC576916) was isolated from Phlebotomus (Paraphlebotomus) mongolensis and Phlebotomus (Pa.) caucasicus, and a new B-group haplotype (AZ2331, GenBank accession JX488735) was isolated from Phlebotomus (Larroussius) perfiliewi. Therefore, Wolbachia was found to occur in at least three of the incriminated vectors of zoonotic cutaneous leishmaniasis and zoonotic visceral leishmaniasis in different geographical regions of Iran. It may provide a new tool for the future control of leishmaniasis.

In this study, we use DNA barcodes for species delimitation to solve taxonomic conflicts in 86 specimens of 14 species belonging to the Corynoneura group (Diptera: Chironomidae: Orthocladiinae), from the Atlantic Forest, Brazil. Molecular analysis of cytochrome c-oxidase subunit I (COI) gene sequences supported 14 cohesive species groups, of which two similar groups were subsequently associated with morphological variation at the pupal stage. Eleven species previously described based on morphological criteria were linked to DNA markers. Furthermore, there is the possibility that there may be cryptic species within the Corynoneura group, since one group of species presented internal grouping, although no morphological divergence was observed. Our results support DNA-barcoding as an excellent tool for species delimitation in groups where taxonomy by means of morphology is difficult or even impossible.

Components of male seminal fluids are known to stimulate fecundity and fertility in females of numerous insect species and paternal effects on offspring phenotype are also known, but no studies have yet demonstrated links between male effects on female reproduction and those on progeny phenotype. In separate laboratory experiments employing 10-day-old virgin females of Cheilomenes sexmaculata (F.), we varied male age and mating history to manipulate levels of male allomones and found that the magnitude of paternal effects on progeny phenotype was correlated with stimulation of female reproduction. Older virgin males remained in copula longer than younger ones, induced higher levels of female fecundity, and sired progeny that developed faster to yield heavier adults. When male age was held constant (13 days), egg fertility declined as a function of previous male copulations, progeny developmental times increased, and the adult weight of daughters declined. These results suggest that male epigenetic effects on progeny phenotype act in concert with female reproductive stimulation; both categories of effects increased as a consequence of male celibacy (factor accumulation), and diminished as a function of previous matings (factor depletion). Male factors that influence female reproduction are implicated in sexual conflict and parental effects may extend this conflict to offspring phenotype. Whereas mothers control the timing of oviposition events and can use maternal effects to tailor progeny phenotypes to prevailing or anticipated conditions, fathers cannot. Since females remate and dilute paternity in polyandrous systems, paternal fitness will be increased by linking paternal effects to female fecundity stimulation, so that more benefits accrue to the male's own progeny.

Oligonychus punicae and Oligonychus perseae (Acari: Tetranychidae) are the most important mite species affecting avocado orchards in Mexico. Here we used nucleotide sequence data from segments of the nuclear ribosomal internal transcribed spacers (ITS1 and ITS2) and mitochondrial cytochrome oxidase subunit I (COI) genes to assess the phylogenetic relationships between both sympatric mite species and, using only ITS sequence data, examine genetic variation and population structure in both species, to test the hypothesis that, although both species co-occur, their genetic population structures are different in both Michoacan state (main producer) and Mexico state. Phylogenetic analysis showed a clear separation between both species using ITS and COI sequence information. Haplotype network analysis done on 24 samples of O. punicae revealed low genetic diversity with only three haplotypes found but a significant geographical population structure confirmed by analysis of molecular variance (AMOVA) and Kimura-2-parameter (K2P) analyses. In addition, a Mantel test revealed that geographical isolation was a factor responsible for the genetic differentiation. In contrast, analyses of 22 samples of O. perseae revealed high genetic diversity with 15 haplotypes found but no geographical structure confirmed by the AMOVA, K2P and Mantel test analyses. We have suggested that geographical separation is one of the most important factors driving genetic variation, but that it affected each species differently. The role of the ecology of these species on our results, and the importance of our findings in the development of monitoring and control strategies are discussed.

Whiteflies have distinct nymphal stages: their first stage is mobile, whereas the later immature stages are sessile. The developmental and structural changes of antennae and antennal sensilla in whiteflies during these stages have rarely been investigated. This paper describes the morphology of antennae and antennal sensilla in four nymphal stages of Aleurodicus dispersus based on scanning electron microscopy. There were significant differences found in shape and length of the antennae, and differences in type, number, morphological structure and distributional pattern of antennal sensilla in the four nymphal stages of A. dispersus. We found two types of sensilla on the antennae of first-instar nymph, three types on the third-instar nymphal antennae, four types on the second-instar and seven types on the fourth-instar nymphal antennae. Sensilla trichoidea (ST) and elevated sensilla placodea were found on the antennae of each nymphal stage, sensilla chaetica only occurred on the antennae of fourth-instar nymph. Sensilla furcatea occurred on the antennae of second- and third-instar nymphs, and sensilla basiconica were found on the antennae of second- and fourth-instar nymphs. In addition, there were sensilla campaniform and sensilla coeloconica found only on the antennae of fourth-instar nymph, whereas the ST of fourth-instar nymphs included sensilla trichoidea 1 and sensilla trichoidea 2. The possible functions of antennal sensilla are discussed. Our results contribute to a better understanding of the development of the olfactory system of whitefly nymphal stages, and provide a basis for further exploration of chemical communication mechanisms between whiteflies and host plants.

The tachinid fly Exorista larvarum (L.) (Diptera: Tachinidae) is a polyphagous larval endoparasitoid that deposits its eggs on the host exoskeleton of lepidopteran and tenthredinid larvae. The attachment of larval E. larvarum and the formation of the respiratory funnel were studied during infestation in the last larval instar of the wax moth, Galleria mellonella (L.) (Lepidoptera: Pyralidae). The tachinid larvae burrow through the host integument after hatching, using their robust cephalopharyngeal skeleton, leaving a dark spot at the point of their penetration as a result of host cuticle melanization. Endoparasitoid penetration induces the host cellular defence, resulting in the formation of a haemocyte capsule consisting of multi-cellular sheaths. This enveloping capsule later undergoes melanization, which is mostly obvious towards the posterior part of the endoparasitoid. The endoparasitoid uses the host encapsulation response to build a respiratory funnel from the modified host integument, leading to the host surface. The encapsulated larva remains attached to the respiratory funnel via an anal hook and cuticular spines until fully developed. Additional immunohistochemical analyses were used to study host–parasitoid interactions. Indirect immunofluorescence showed no labelling of potential tachinid antigens and confirmed no effect on the surrounding host tissues. A simulated parasitization with coated polybead microspheres revealed the mortal impact of tachinid antigens to the host. Hosts injected with antigen-coated polybeads died as a consequence of an acute and extensive immunological response to the tachinid antigens and not due to the trauma caused by foreign objects inside their body.

Laboratory studies were conducted to assess the effect of temperature on the survival, development, longevity and fecundity of Helicoverpa armigera (Lepidoptera: Noctuidae) at eight different fluctuating temperatures with an amplitude ±9 °C under constant photoperiodic conditions of 16:8 h (L:D). H. armigera achieved complete development from egg to adult emergence between mean 17.5 and 32.5 °C. At mean 35 °C, all newly hatched larvae died and at mean 15 °C entered diapause at pupal stage. The lower developmental thresholds of the immature stages were estimated by a linear model and ranged from 4.63 °C (pupal stage) to 7.69 °C (egg stage). The developmental thresholds estimated by a nonlinear model were slightly higher than those estimated by the linear model. Adult longevity and fecundity were reduced at mean fluctuating temperatures 17.5 and 32.5 °C, but tended to be independent of the pattern of temperature change at moderate temperatures. The maximum reproductive performance, 1130 eggs per female, was observed at mean 25 °C. The intrinsic rates of increase were positive, meaning that H. armigera could be expected to persist or increase in number between mean 17.5 and 32.5 °C, with the maximum value at mean 27.5 °C. H. armigera survives, develops and reproduces within a wide range of fluctuating temperatures, while it completes the above functions with different levels of success at different mean temperatures of diurnal variation. Comparison of our results with similar data from the literature involving constant conditions is discussed. This information will provide a better understanding of H. armigera phenology and population dynamics under natural conditions and is essential to understanding the ecological and evolutionary consequences of climate change on this important species.

The rice stem bug, Tibraca limbativentris Stal. (Hemiptera: Pentatomidae) is one of the most important pests of rice crops, especially irrigated crops. Plant defence strategies against these bugs may involve the emission of chemical compounds, which are released following herbivore attacks, directly or indirectly harming pest performance. The aim of this study was to evaluate the influence of constitutive and herbivory-induced volatiles from rice plants (Oryza sativa L.) on the behavioural responses of T. limbativentris adults and egg parasitoids Trissolcus basalis (Wollaston) and Telenomus podisi (Ashmead) (Hymenoptera, Platygastridae). Plant volatiles were collected from undamaged plants of the rice cultivar IRGA 424 and from plants that suffered herbivory by five males or five females of T. limbativentris. Air-entrainment extracts were analysed by GC–flame ionization detector and GC–MS, and insect responses evaluated in a ‘Y’ olfactometer. T. limbativentris feeding damaged on rice plants induced the release of 16 volatiles compounds in a higher amounts compared to undamaged plants The main compounds induced were (E)-2-hexenal, (E)-2-octen-1-ol, methyl salicylate and α-muurolene. Female bugs were significantly attracted to air-entrainment extracts containing volatiles from undamaged plants compared with air-entrainment extracts containing volatiles emitted from plants damaged by T. limbativentris, whereas males showed no preference. Telenomus podisi females were significantly attracted to volatiles from air-entrainment extracts of plants damaged by females, whereas T. basalis showed no preference. These results suggest that rice plants may be emitting defence compounds, which could be avoided by T. limbativentris females and also acted indirectly by attracting natural enemies.

Several species of the genus Cryptolestes Ganglbauer, 1899 (Coleoptera: Laemophloeidae) are commonly found in stored products. In this study, five species of Cryptolestes, with almost worldwide distribution, were obtained from laboratories in China, Czech Republic and the USA: Cryptolestes ferrugineus (Stephens, 1831), Cryptolestes pusillus (Schönherr, 1817), Cryptolestes turcicus (Grouvelle, 1876), Cryptolestes pusilloides (Steel & Howe, 1952) and Cryptolestes capensis (Waltl, 1834). Molecular identification based on a 658 bp fragment from the mitochondrial DNA cytochrome c oxidase subunit I (COI) was adopted to overcome some problems of morphological identification of Cryptolestes species. The utility of COI sequences as DNA barcodes in discriminating the five Cryptolestes species was evaluated on adults and larvae by analysing Kimura 2-parameter distances, phylogenetic tree and haplotype networks. The results showed that molecular approaches based on DNA barcodes were able to accurately identify these species. This is the first study using DNA barcoding to identify Cryptolestes species and the gathered DNA sequences will complement the biological barcode database.

Natural enemies that respond to prey in a density-dependent manner may be able to quickly suppress pest populations before they reach economically damaging levels. Although it is primarily the combination of a natural enemy's functional response and a population numerical response that will influence the maximum number of pests attacked, other factors may influence a density-dependent response. We conducted large-scale field experiments, both artificially inoculating grapevines with larvae and using naturally occurring populations, to quantify and characterize the response of a parasitoid, Dolichogenidea tasmanica (Cameron) (Hymenoptera: Braconidae) to different densities of its host, the pest of grapevines, Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae). We showed that the response of D. tasmanica to the density of E. postvittana was inversely density-dependent, and that the degree of parasitism was consistently and significantly higher in the grape variety Cabernet Sauvignon compared with Chardonnay. While the significant effect of variety on the degree of parasitism may provide an option for increasing the parasitism of E. postvittana by D. tasmanica, it also highlights how differences in host plant can influence trophic interactions.

Understanding the biology of the housefly (Musca domestica L.) is crucial for the development of mass-rearing protocols in order to use this insect as a degradation agent for livestock waste. In this study, the biological and genetic differences between different laboratory strains of M. domestica were analysed. Additionally, hybrids were obtained by mixing the strains and their biological parameters were also measured. The three strains of M. domestica presented differences in their biological and morphological parameters, the main differences were: size, egg production and developmental time. The strain A (specimens from Central Europe) had the best qualities to be used in mass-rearing conditions: it produced the largest quantities of eggs (5.77±0.38 eggs per female per day), the individuals were larger (12.62±0.22 mg) and its developmental time was shorter (15.22±0.21 days). However, the strain C (specimens from SW Europe) produced the fewest eggs (3.15±0.42 eggs per female per day) and needed 18.16±0.49 days to develop from larva to adult, whilst the females from strain B (from South America) produced 4.25±0.47 eggs per day and needed 17.11±0.36 days to complete its development. Genetic analysis of the original laboratory strains showed four different mtDNA cytochrome c oxidase subunit I haplotypes. Statistical parsimony network analysis showed that the SW Europe and South-American strains shared haplotypes, whereas the Central Europe strain did not. Upon hybridizing the strains, variations in egg production and in developmental time were observed in between hybrids and pure strains, and when mixing Central European and South-American strains only males were obtained.

For a better understanding of the population structure and dispersal rates of Sogatella furcifera, we developed 21 novel polymorphic expressed sequence tags (EST) derived microsatellites, which were successfully amplified in four multiplex polymerase chain reaction sets. These new microsatellites were firstly assessed in 20 individuals sampled from Wenshan in China. The results showed that all 21 loci were highly polymorphic; the number of alleles ranged from 3 to 9, with an average of 4.8 alleles per locus. The observed and expected heterozygosity ranged from 0.200 to 0.900 and from 0.184 to 0.799, respectively. Nineteen of the 21 microsatellites without null allele, were subsequently used for population genetic structure analyses of five S. furcifera populations sampled in south region of China (sites up to 1314 kilometers apart). The observed and expected heterozygosity for each population ranged from 0.436 to 0.494 and from 0.454 to 0.482, respectively. The level of population differentiation was very low, with an average pairwise FST of 0.002. Bayesian cluster analysis result suggested that the five S. furcifera populations formed one genetic cluster. Discriminant analysis of principle components detected three genetic clusters. The spread of the three clusters across the five populations explained the lack of population differentiation and the Bayesian cluster result. All the results indicated that long-distance migration of this pest allowed genetic mixing between populations from remote geographical origins. These new microsatellites will be powerful tools for population genetics studies of S. furcifera.

Plutella xylostella (L.) is the most important pest of Brassicaceae worldwide, with a recent estimate of US$ 4–5 billion expenditure for the control of this insect. A case of very high resistance of this pest to chlorantraniliprole was recently associated with reduced efficacy in a Brazilian field of Brassica spp. Although diamide resistance has been characterized, the fitness of insects due to such resistance has yet to be examined. Therefore, in this study, biological parameters were assessed in both susceptible and resistant strains of P. xylostella subjected to sublethal chlorantraniliprole concentrations. The field strain showed high resistance to chlorantraniliprole (RR50=27,793-fold), although resistance rapidly decreased in the first generations, showing instability. The exposure of susceptible and resistant larvae to their respective LC1, LC10, and LC25 values led to an increased duration of the larval and pupae phases and reduced weight in both strains; however, no significant differences in pupal viability across the treatments were observed. The resistant insects presented significantly lower larval weight and fecundity and higher larval and pupal periods, hatchability, and male longevity when not exposed to chlorantraniliprole, suggesting a fitness cost associated with resistance. In addition, resistant females showed a significantly higher egg-laying period and longevity at LC25, whereas the males lived longer at LC1. Chlorantraniliprole negatively impacted the biological parameters of both strains tested, although these effects were more relevant to the resistant insects. Resistant P. xylostella showed negative and positive biological trade-offs when compared with the susceptible individuals in both the absence and presence of chlorantraniliprole. Despite the important role that these trade-offs may play in the evolution of resistance to chlorantraniliprole, practical applications still depend on such information as the dominance of fitness costs and resistance.

Discrimination of particular species within the species complexes of tephritid fruit flies is a very challenging task. In this fruit-fly family, several complexes of cryptic species have been reported, including the African cryptic species complex (FAR complex). Cuticular hydrocarbons (CHCs) appear to be an excellent tool for chemotaxonomical discrimination of these cryptic species. In the present study, CHC profiles have been used to discriminate among three important agricultural pests from the FAR complex, Ceratitis fasciventris, Ceratitis anonae and Ceratitis rosa. Hexane body surface extracts of mature males and females were analyzed by two-dimensional gas chromatography with mass spectrometric detection and differences in CHC profiles between species and sexes tested through multivariate statistics and compared with species identification by means of microsatellite markers. Quantitative as well as qualitative CHC profile differences between sexes and species are reported. The CHC profiles consisted of a mixture of linear, internally methyl-branched and mono-, di- and tri-unsaturated alkanes. Twelve compounds were pinpointed as potential chemotaxonomical markers. The present study shows that presence or absence of particular CHCs might be used in the chemical diagnosis of the FAR complex. Moreover, our results represent an important first step in the development of a useful chemotaxonomic tool for cryptic species identification of these important agricultural pests.

The greenhouse whitefly Trialeurodes vaporariorum Westwood, 1856 (Hemiptera: Aleyrodidae) is an invasive and highly polyphagous phloem-feeding pest of vegetables and ornamentals. Trialeurodes vaporariorum causes serious damage due to direct feeding and transmits several important plant viruses. Excessive use of insecticides has resulted in significantly reduced levels of susceptibility of various T. vaporariorum populations. To determine the genetic variability within and among populations of T. vaporariorum from Serbia and to explore their genetic relatedness with other T. vaporariorum populations, we analysed the mitochondrial cytochrome c oxidase I (COI) sequences of 16 populations from Serbia and six neighbouring countries: Montenegro (three populations), Macedonia (one population) and Croatia (two populations), for a total of 198 analysed specimens. A low overall level of sequence divergence and only five variable nucleotides and six haplotypes were found. The most frequent haplotype, H1, was identified in all Serbian populations and in all specimens from distant localities in Croatia and Macedonia. The COI sequence data that was retrieved from GenBank and the data from our study indicated that H1 is the most globally widespread T. vaporariorum haplotype. A lack of spatial genetic structure among the studied T. vaporariorum populations, as well as two demographic tests that we performed (Tajima's D value and Fu's Fs statistics), indicate a recent colonisation event and population growth. Phylogenetic analyses of the COI haplotypes in this study and other T. vaporariorum haplotypes that were retrieved from GenBank were performed using Bayesian inference and median-joining (MJ) network analysis. Two major haplogroups with only a single unique nucleotide difference were found: haplogroup 1 (containing the five Serbian haplotypes and those previously identified in India, China, the Netherlands, the United Kingdom, Morocco, Reunion and the USA) and haplogroup 3 (containing the single Serbian haplotype H3 and haplotypes from Costa Rica, the USA and Spanish Canary Islands). Collectively, our data indicate a rather limited value of COI as a genetic marker for discrimination between different T. vaporariorum populations in the investigated area. Possible explanations for the observed lack of COI sequence variability, such as specific genetics of biological invasion and/or the influence of bacterial symbionts that manipulate insect reproduction, are discussed.

Mealybugs (Hemiptera: Pseudococcidae) are important pests for crops worldwide. Different species, cryptic taxa under the same species name or even populations within a species can differ in biological characteristics, such as phenology, resistance to insecticides, virus transmission and susceptibility to natural enemies. Therefore, their management efficacy depends on their accurate identification. Microsatellite genetic markers are efficient in revealing the fine-scale taxonomic status of insects, both at inter- and intra-specific level. Despite their potential uses, microsatellites have been developed only for one mealybug species so far. Hence, it is unclear whether microsatellites may be useful to assess mealybug population differentiation and structuring. In this work, we tested the feasibility of developing microsatellite markers in mealybugs by: (i) producing and characterizing microsatellite DNA libraries for three species: Pseudococcus viburni, Pseudococcus comstocki and Heliococcus bohemicus, and (ii) by developing and testing markers for Ps. viburni. The obtained libraries contained balanced percentages of dinucleotide (ranging from 15 to 25%) and trinucleotide (from 5 to 17%) motifs. The marker setup for Ps. viburni was successful, although 70% of the primers initially tested were discarded for a lack of polymorphism. Finally, 25 markers were combined in two multiplex polymerase chain reactions with 21 displaying no evidence of deviation from Hardy–Weinberg equilibrium. Ps. viburni markers were tested on one population from France and one from Chile. The markers revealed a significant genetic differentiation between the two populations with an Fst estimate of 0.266.