Mating behavior of Pseudococcus calceolariae and Pseudococcus longispinus (Hemiptera: Pseudococcidae): are asexual reproduction and hybridization possible?

The study of insect reproduction is important from both basic and applied perspectives, particularly in mealybugs (Hemiptera: Pseudococcidae), because of the diversity of reproduction modes and also because they are important agricultural pests. Sex pheromone control strategies are currently being developed for many species. Pseudococcus calceolariae (Maskell) and Pseudococcus longispinus (Targioni Tozzetti) are closely related species that often coexist in the same host plant. In this study, mating behavior, the possible occurrence of asexual reproduction, and hybridization between them were investigated. We confirmed that both species did not show asexual reproduction and required the presence of a male to reproduce. When couples of the same species were put together, males had a highly stereotyped mating behavior, and females showed an active role in mating success by accepting or rejecting males with abdominal movements. In hybridization trials, no progeny was obtained for any of the interspecific combinations. Moreover, in interspecific pairs, males mainly moved randomly in the arena without direct contact with females and females showed no willingness to mate, escape, or not move in the presence of the male. Therefore, courtship and copulation success in both species were directly related to the specificity of the mating pair and, there was no evidence of hybridization. This information is useful for the understanding of reproduction in this family and supports the development of management techniques based on sex pheromones to disrupt reproduction or to monitor these mealybug species populations.

Conservation biological control of arthropod pests using native plants.

Habitat or vegetation manipulation is a strategy within conservation biological control, yet, its formal study is relatively recent and lacking in many parts of the world, particularly when using native plants. We found 56 studies dealing with native plants and conservation biological control, from 2002 to 2022. Most were carried out in developed countries, and studied the provision of flower resources to natural enemies' complexes in several crops. The main gaps in knowledge, similar to studies with non-native plants, were the lack of understanding mechanisms, unknown effects on pest populations, and absence of cost-benefit analyses. Breaching these gaps will allow this strategy to be more transferable among cropping systems and locations and will promote its use by farmers.

A roadmap for ladybird conservation and recovery.

Ladybirds (Coleoptera: Coccinellidae) provide services that are critical to food production, and they fulfill an ecological role as a food source for predators. The richness, abundance, and distribution of ladybirds, however, are compromised by many anthropogenic threats. Meanwhile, a lack of knowledge of the conservation status of most species and the factors driving their population dynamics hinders the development and implementation of conservation strategies for ladybirds. We conducted a review of the literature on the ecology, diversity, and conservation of ladybirds to identify their key ecological threats. Ladybird populations are most affected by climate factors, landscape composition, and biological invasions. We suggest mitigating actions for ladybird conservation and recovery. Short-term actions include citizen science programs and education, protective measures for habitat recovery and threatened species, prevention of the introduction of non-native species, and the maintenance and restoration of natural areas and landscape heterogeneity. Mid-term actions involve the analysis of data from monitoring programs and insect collections to disentangle the effect of different threats to ladybird populations, understand habitat use by taxa on which there is limited knowledge, and quantify temporal trends of abundance, diversity, and biomass along a management-intensity gradient. Long-term actions include the development of a worldwide monitoring program based on standardized sampling to fill data gaps, increase explanatory power, streamline analyses, and facilitate global collaborations.

Las catarinas (Coleoptera: Coccinellidae) proporcionan servicios que son críticos para la producción de alimento, y juegan un papel ecológico como fuente de alimento para depredadores. Sin embargo, la riqueza, abundancia y distribución de catarinas están en peligro debido a muchas amenazas antropogénicas. La carencia de conocimiento sobre el estatus de conservación de la mayoría de las especies y los factores que inciden en su dinámica poblacional dificulta el desarrollo e implementación de estrategias de conservación para las catarinas. Realizamos una revisión de la literatura sobre la ecología, diversidad y conservación de catarinas para identificar sus amenazas ecológicas clave. Las poblaciones de catarinas fueron afectadas mayormente por factores climáticos, composición del paisaje e invasiones biológicas. Proponemos acciones de mitigación para la conservación y recuperación de catarinas. Acciones a corto plazo incluyen programas de ciencia y educación ciudadana, medidas de protección para la recuperación de hábitat y de especies amenazadas, prevención de la introducción de especies no nativas y el mantenimiento y restauración de áreas naturales y la heterogeneidad del paisaje. Acciones a mediano plazo implican el análisis de datos obtenidos de programas de monitoreo y colecciones de insectos para desenmarañar el efecto de las diferentes amenazas a las poblaciones de catarinas, comprender el uso del hábitat por taxa de los que se tiene conocimiento limitado y cuantifica las tendencias temporales de la abundancia, diversidad y biomasa a lo largo de un gradiente de intensidad de manejo. Acciones a largo plazo incluyen el desarrollo de un programa de monitoreo a nivel mundial basado en muestreos estandarizados para subsanar la falta de datos, incrementar el poder explicativo, optimizar los análisis y facilitar colaboraciones globales.

Citizen Science and Phytosanitary Surveillance Systems Are Complementary Tools to Follow the Invasion of Harmonia axyridis.

Citizen science is a valuable tool for early detection, distribution, and spread of invasive alien species (IAS). Nevertheless, citizen science initiatives have several potential biases and may be complemented with long-term structured monitoring schemes. We analyzed the spatial-temporal dynamics of the invasion of Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) in Chile, based upon two citizen sciences databases (WEB and INAT) and one structured monitoring (SAG). We collected 8638 H. axyridis occurrences between 2009 and 2020. WEB had a higher number of records than SAG and INAT, and in all databases, the number of records has increased over time. The three databases showed that the invasion started in central Chile and then spread toward the north and south. The WEB and SAG recorded occurrences in the extreme north and south, whereas INAT concentrated all the occurrences in a more limited area, included in WEB and SAG. Both citizen science initiatives concentrated their records in areas of high human populations whereas SAG records had a more even distribution across regions. At 2020, WEB accounted for 55%, SAG 54%, and INAT 8% of the total area accumulated with H. axyridis, with only 16% of area shared among databases. WEB and INAT obtained most of their records in urban and industrial land cover types, while SAG records were more evenly represented in different land cover types. Our results confirm that combined methods, including citizen science initiatives, national surveillance system, and localized samplings, complement each other in providing knowledge to understand the patterns, processes, and consequences of this invasion.

Mating Disruption of Pseudococcus calceolariae (Maskell) (Hemiptera, Pseudococcidae) in Fruit Crops.

Pseudococcus calceolariae, the citrophilous mealybug, is a species of economic importance. Mating disruption (MD) is a potential control tool. During 2017-2020, trials were conducted to evaluate the potential of P. calceolariae MD in an apple and a tangerine orchard. Two pheromone doses, 6.32 g/ha (2017-2018) and 9.45 g/ha (2019-2020), were tested. The intermediate season (2018-2019) was evaluated without pheromone renewal to study the persistence of the pheromone effect. Male captures in pheromone traps, mealybug population/plant, percentage of infested fruit at harvest and mating disruption index (MDI) were recorded regularly. In both orchards, in the first season, male captures were significantly lower in MD plots compared to control plots, with an MDI > 94% in the first month after pheromone deployment. During the second season, significantly lower male captures in MD plots were still observed, with an average MDI of 80%. At the third season, male captures were again significant lower in MD than control plots shortly after pheromone applications. In both orchards, population by visual inspection and infested fruits were very low, without differences between MD and control plots. These results show the potential use of mating disruption for the control of P. calceolariae.

Natural enemies of armored scales (Hemiptera: Diaspididae) and soft scales (Hemiptera: Coccidae) in Chile: Molecular and morphological identification.

Scale insects (Hemiptera: Sternorrhyncha: Coccomorpha) are key pests of agricultural crops and ornamental plants worldwide. Their populations are difficult to control, even with insecticides, due to their cryptic habits. Moreover, there is growing concern over the use of synthetic pesticides for their control, due to deleterious environmental effects and the emergence of resistant populations of target pests. In this context, biological control may be an effective and sustainable approach. Hymenoptera Chalcidoidea includes natural enemies of scale insects that have been successfully used in many biological control programs. However, the correct identification of pest scale species and their natural enemies is particularly challenging because these insects are very small and highly specialized. Integrative taxonomy, coupling DNA barcoding and morphological analysis, has been successfully used to characterize pests and natural enemy species. In this study, we performed a survey of parasitoids and predators of armored and soft scales in Chile, based on 28S and COI barcodes. Fifty-three populations of Diaspididae and 79 populations of Coccidae were sampled over the entire length of the country, from Arica (18°S) to Frutillar (41°S), between January 2015 and February 2016. The phylogenetic relationships obtained by Bayesian inference from multilocus haplotypes revealed 41 putative species of Chalcidoidea, five Coccinellidae and three Neuroptera. Species delimitation was confirmed using ABGD, GMYC and PTP model. In Chalcidoidea, 23 species were identified morphologically, resulting in new COI barcodes for 12 species and new 28S barcodes for 14 species. Two predator species (Rhyzobius lophantae and Coccidophilus transandinus) were identified morphologically, and two parasitoid species, Chartocerus niger and Signiphora bifasciata, were recorded for the first time in Chile.

Synthesis of citrophilus mealybug sex pheromone using chrysanthemol extracted from Pyrethrum ( Tanacetum cinerariifolium).

A commercial pyrethrum extract was used as a source of chrysanthemol for the synthesis of the citrophilus mealybug ( Pseudococcus calceolariae) sex pheromone. The chrysanthemic acid esters (pyrethrins I) were isolated and subsequently reduced to obtain chrysanthemol, which was used for ester pheromone synthesis. Field tests showed that the pheromone synthesized using plant-derived chrysanthemol was as attractive to male P. calceolariae as the pheromone obtained using a commercial isomeric chrysanthemol mixture.

Effects of inbreeding on a gregarious parasitoid wasp with complementary sex determination.

Inbreeding and inbreeding depression are processes in small populations of particular interest for a range of human activities such as animal breeding, species conservation, or pest management. In particular, biological control programs should benefit from a thorough understanding of the causes and consequences of inbreeding because natural enemies experience repetitive bottlenecks during importation, laboratory rearing, and introduction. Predicting the effect of inbreeding in hymenopteran parasitoid wasps, frequently used in biological control programs, is nonetheless a difficult endeavor. In haplodiploid parasitoids, the purge of deleterious alleles via haploid males should reduce genetic load, but if these species also have complementary sex determination (CSD), abnormal diploid males will be produced, which may jeopardize the success of biological control introductions. Mastrus ridens is such a parasitoid wasp with CSD, introduced to control the codling moth, Cydia pomonella (L.). We studied its life history traits in the laboratory under two conditions: inbred (full-sib) and outbred (nonsib) crosses, across five generations, to examine the consequences of inbreeding in this species. We found that in inbred lines, nonreproducing females live less, the number of daughters produced was lower, and sex ratio (proportion of males) and proportion of diploid males were higher. Diploid males were able to produce fertile daughters, but fewer than haploid males. Lineage survival was similar for inbred and outbred lines across the five generations. The most significant decrease in fitness was thus a consequence of the production of diploid males, but this effect was not as extreme as in most other species with CSD, due to the fertility of diploid males. This study highlights the importance of determining the type of sex determination in parasitoid wasps used for biological control, and the importance of maintaining genetic diversity in species with CSD when importation or augmentation is the goal.

Searching Behavior of Cryptolaemus montrouzieri (Coleoptera: Coccinellidae) in Response to Mealybug Sex Pheromones.

Cryptolaemus montrouzieri (Mulsant) (Coleoptera: Coccinellidae) is a predatory insect widely commercialized for the augmentative biological control of mealybugs (Hemiptera: Pseudococcidae). Though visual factors that guide this predator to its prey have been reported, the chemical cues that they may also use have not been well studied. In this study, the response of C. montrouzieri to synthetic sex pheromone of two mealybug species, Pseudococcus calceolariae (Maskell) (Hemiptera: Pseudococcidae) and Pseudococcus viburni (Signoret) (Hemiptera: Pseudococcidae), were assessed through Y-tube olfactometer and characterization of searching behavior with a video-tracking system. In the olfactometer, both C. montrouzieri males and females showed a significant response to the pheromones of both mealybug species separately. The searching behavior differs depending on the pheromone species, with a shorter total distance, reduction of mean velocity and higher meander with the pheromone stimulus of P. calceolariae when compared to P. viburni. Therefore, it is shown that this predator responds to the pheromone stimuli of its prey, and can do it in a species-specific way. These results can have a practical application through the use of pest pheromones to attract predators to a crop as part of an integrated pest management program.

Landscape crop composition effects on cotton yield, Lygus hesperus densities and pesticide use.

BACKGROUND: Landscape crop composition surrounding agricultural fields is known to affect the density of crop pests, but quantifying these effects, as well as measuring how they translate to changes in yield, is difficult. Using a large dataset consisting of 1498 records of commercial cotton production in California between 1997 and 2008, we explored the relationship between landscape composition and cotton yield, the density of Lygus hesperus (a key cotton pest) at field-level and within-field spatial scales and pesticide use. RESULTS: We found that the crop composition immediately adjacent to a cotton field was associated with substantial differences in cotton yield, L. hesperus density and pesticide use. Furthermore, crops that tended to be associated with increased L. hesperus density also tended to be associated with increased pesticide use and decreased cotton yield. CONCLUSION: Our results suggest a possible mechanism by which landscape composition can affect cotton yield: by increasing the density of pests which in turn damage cotton plants. Our quantification of how surrounding crops affect pest densities, and in turn yield, in cotton fields has significant impacts for cotton farmers, who can use this information to help optimize crop selection and ranch layout. © 2016 Society of Chemical Industry.

Mealybug species from Chilean agricultural landscapes and main factors influencing the genetic structure of Pseudococcus viburni.

The present study aimed to characterize the distribution of mealybug species along Chilean agro-ecosystems and to determine the relative impact of host plant, management strategy, geography and micro-environment on shaping the distribution and genetic structure of the obscure mealybug Pseudococcus viburni. An extensive survey was completed using DNA barcoding methods to identify Chilean mealybugs to the species level. Moreover, a fine-scale study of Ps. viburni genetic diversity and population structure was carried out, genotyping 529 Ps. viburni individuals with 21 microsatellite markers. Samples from 16 localities were analyzed using Bayesian and spatially-explicit methods and the genetic dataset was confronted to host-plant, management and environmental data. Chilean crops were found to be infested by Ps. viburni, Pseudococcus meridionalis, Pseudococcus longispinus and Planococcus citri, with Ps. viburni and Ps. meridionalis showing contrasting distribution and host-plant preference patterns. Ps. viburni samples presented low genetic diversity levels but high genetic differentiation. While no significant genetic variance could be assigned to host-plant or management strategy, climate and geography were found to correlate significantly with genetic differentiation levels. The genetic characterization of Ps. viburni within Chile will contribute to future studies tracing back the origin and improving the management of this worldwide invader.

Monitoring Pseudococcus calceolariae (Hemiptera: Pseudococcidae) in Fruit Crops Using Pheromone-Baited Traps.

The citrophilus mealybug, Pseudococcus calceolariae (Maskell), is an important pest of fruit crops in many regions of the world. Recently, its sex pheromone has been identified and synthesized. We carried out field experiments with the goal of developing monitoring protocols for P. calceolariae using pheromone-baited traps. Traps checked hourly for 24 hours showed a distinct diel pattern of male flight, between 18:00 and 21:00 h. The presence of unnatural stereoisomers did not affect trap captures, with isomeric mixtures capturing similar amounts of males as the biological active isomer. Dose of isomeric mixture pheromone (0-100 µg) had a nonlinear effect on male captures, with 10, 30, and 50 µg capturing similar amounts. The effective range of pheromone traps was determined by placing traps at different distances (15, 40, and 80 m) from an infested blueberry field, loaded with 0, 1 and 25 µg of the pheromone. For all distances, 25 µg dose captured more males, and was highly attractive up to 40 m. There was a significant effect of lure age on male captures (0-150 d), with similar amount of males captured up to 90-day-old lure, and lower captures in the 150-day-old lure compared with fresh ones. We found significant positive correlations between P. calceolariae males caught in pheromone traps with female abundance and fruit infestation at harvest. Our results show the usefulness of P. calceolariae pheromones for monitoring at field level and provide information for the design of monitoring protocols.

Development of microsatellite markers and detection of genetic variation between Goniozus wasp populations.

Molecular genetic markers reveal differences between genotypes according to the presence of alleles (the same or different) at target loci. Microsatellite markers are especially useful co-dominant markers that have been used in a wide range of studies to elucidate the population structure and dynamics of a range of organisms, including agriculturally beneficial insects such as parasitic wasps (parasitoids). In the present study, twelve primer pairs were designed for the south Asian , Goniozus nephantidis (Muesebeck) (Hymenoptera: Bethylidae), and 24 for its New World congener, Goniozus legneri Gordh, parasitoids of the larvae of the lepidopteran coconut pest Opisina arenosella Walker (Lepidoptera: Crytophasidae) and other lepidopteran pests, respectively, in order to investigate polymorphism within and between populations. The wasps fingerprinted were a total of 85 G. nephantidis and G. legneri, including individuals belonging to three putatively different strains of G. legneri. Annealing gradient tests (50-65°C) were conducted to study the quality of the PCR amplification across an annealing temperature gradient using a mixed genotype DNA template from each species separately. Seven primer pairs, which amplified clear products of approximately the expected size of G. nephantidis and 18 of G. legneri, were then selected for capillary analysis for fragment size determination on a Beckmann CEQ 8000. Neither G. nephantidis nor G. legneri were polymorphic within populations. However, there were six primer pairs that did show polymorphism between G. legneri populations that originated from different geographical areas within South America (Uruguay and Chile). Furthermore, one primer pair revealed diversity between the two strains collected within Chile. One of the markers was subsequently used to provide unbiased assessment of primary sex ratio in G. legneri.

Ecoinformatics for integrated pest management: expanding the applied insect ecologist's tool-kit.

Experimentation has been the cornerstone of much of integrated pest management (IPM) research. Here, we aim to open a discussion on the possible merits of expanding the use of observational studies, and in particular the use of data from farmers or private pest management consultants in "ecoinformatics" studies, as tools that might complement traditional, experimental research. The manifold advantages of experimentation are widely appreciated: experiments provide definitive inferences regarding causal relationships between key variables, can produce uniform and high-quality data sets, and are highly flexible in the treatments that can be evaluated. Perhaps less widely considered, however, are the possible disadvantages of experimental research. Using the yield-impact study to focus the discussion, we address some reasons why observational or ecoinformatics approaches might be attractive as complements to experimentation. A survey of the literature suggests that many contemporary yield-impact studies lack sufficient statistical power to resolve the small, but economically important, effects on crop yield that shape pest management decision-making by farmers. Ecoinformatics-based data sets can be substantially larger than experimental data sets and therefore hold out the promise of enhanced power. Ecoinformatics approaches also address problems at the spatial and temporal scales at which farming is conducted, can achieve higher levels of "external validity," and can allow researchers to efficiently screen many variables during the initial, exploratory phases of research projects. Experimental, observational, and ecoinformatics-based approaches may, if used together, provide more efficient solutions to problems in pest management than can any single approach, used in isolation.

The absolute configuration of the sex pheromone of the citrophilous mealybug, Pseudococcus calceolariae.

The absolute configuration of the sex pheromone of the citrophilous mealybug, Pseudococcus calceolariae, was determined to be (1R,3R)-[2,2-dimethyl-3-(2-methylprop-1-enyl)cyclopropyl]methyl (R)-2-acetoxy-3-methylbutanoate. NMR, derivatization reactions, chiral gas chromatography-mass spectrometry, and comparison with synthetic chiral reference compounds, were used to determine the absolute configuration of this compound. This activity of this compound was further confirmed by testing synthetic stereoisomers of the compound as lures in traps for adult male mealybugs. Traps baited with 1,000 µg of the pheromone compound caught 36 times more males than traps baited with virgin females. A mixture of stereoisomers of the pheromone compound can be used for field trapping without adverse effects on trap catch. A comparison with the structures of other sex pheromones of mealybugs is presented.

Development of a multiplex PCR for identification of vineyard mealybugs.

A simple molecular tool was developed and tested to identify seven mealybug species found in North American vineyards: Pseudococcus maritimus Ehrhorn, Pseudococcus viburni (Signoret), Pseudococcus longispinus (Targioni-Tozzeti), Pseudococcus calceolariae (Maskell), Planococcus ficus (Signoret), Planococcus citri (Risso), and Ferrisia gilli Gullan. The developed multiplex PCR is based on the mitochondrial cytochrome c oxidase subunit one gene. In tests, this single-step multiplex PCR correctly identified 95 of 95 mealybug samples, representing all seven species and collected from diverse geographic regions. To test the sensitivity, single specimen samples with different Pl. ficus developmental stages (egg to adult female and adult male) were processed PCR and the resulting output provided consistent positive identification. To test the utility of this protocol for adult males caught in sex baited pheromone traps, Pl. ficus adult males were placed in pheromone traps, aged at a constant temperature of 26±2°C, and processed with the multiplex each day thereafter for 8 d. Results showed consistent positive identification for up to 6 d (range, 6-8 d). Results are discussed with respect to the usefulness of this molecular tool for the identification of mealybugs in pest management programs and biosecurity of invasive mealybugs.

A transient, positive effect of habitat fragmentation on insect population densities.

We conducted an experimental landscape study to test the hypotheses that: (1) habitat removal results in short-term increases in population density in the remaining habitat patches (the crowding effect); (2) following habitat removal, density is higher in landscapes with more, smaller patches and more habitat edge (i.e., a higher level of habitat fragmentation per se) than in less fragmented landscapes, for the same total amount of habitat on the landscapes; (3) this positive effect of fragmentation per se on density is larger in landscapes with smaller inter-patch distances; and (4) these last two effects should be reduced or disappear over time following habitat removal. Our results did not support the first hypothesis, but they provided some support for the other three hypotheses, for two of the four Coccinellid species studied. As in other empirical studies of fragmentation per se on population density, the effects of fragmentation per se were weak and positive (when they did occur). This is the first study to document a transient effect of fragmentation per se on population density, and to show that this effect depends on inter-patch distances. We suggest that fragmentation per se increased the rate of immigration to patches, resulting in higher population densities in more fragmented landscapes.