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We report here a new species belonging to Nephus (Nephus) Mulsant. Nephus (Nephus) apoloniasp. nov. was collected in the Reunion Island (Mascarene Archipelago, Indian Ocean). We describe this new species and redescribe and illustrate three other Nephus species already known from Reunion: Nephus (Nephus) oblongosignatus Mulsant, 1850, Nephus (Geminosipho) reunioni (Fürsch, 1974) and Nephus (Nephus) voeltzkowi Weise, 1910. Furthermore, we present a phylogenetic tree for these four species and calculate the genetic distances between them, using high-throughput DNA sequencing of the mitochondrial genome. The similar external morphology of N. apoloniasp. nov. and N. voeltzkowi very probably explains why individuals from the first species have been mistakenly identified as the latter and were not recognized as different until now. Other than external and genitalia traits, the present study provides molecular evidence confirming these are indeed two different species.

The ladybird beetle Serangium japonicum is an important predator of whiteflies. Investigations of the molecular mechanisms of this predatory beetle have been hindered by the scarcity of gene sequence data. To obtain gene sequences for the ladybird beetle and determine differences in gene expression between the summer and winter seasons, paired-end sequencing was performed. Real-time PCR was used to validate differences in Krueppel homolog 1 gene (Kr-h1) mRNA expression in summer vs. winter samples. To determined the diversity of the population, annotated cytochrome c oxidase subunit I gene (COX1) gene fragments were amplified from several ladybird beetle populations. The analysis yielded 191,246 assembled unigenes, 127,016 of which (66.4%) were annotated. These functional annotations of gene sequences are currently available from the National Center for Biotechnology Information (NCBI), and will provide a basis for studying the molecular mechanisms underlying the biological characteristics of S. japonicum. We found a change in expression of ribosome-associated genes across seasons, and postulate that this change is because of seasonal variation in temperature and photoperiod. The differential expression of Kr-h1 suggests that S. japonicum can successfully overwinter because the adults enter diapause. To explain the effects of season on Kr-h1 gene expression, we hypothesize a model in which that a short photoperiod affects the density of Ca2+, the subsequent activity of methyl farnesoate epoxidase and the synthesis of JH, and in turn Kr-h1 gene expression. COX1 annotation was concordant with the morphological ID. The same COX1 sequence was found in the samples from several provinces in China. Therefore, the COX1 sequence is worth further study to distinguish beetle species and populations.

Simple Summary The solanum whitefly, Aleurothrixus trachoides, is a polyphagous pest known to attack > 70 crops worldwide. Endemic to the Neotropical region, in the past few years, it has emerged as a significant pest of several horticultural crops including pepper in the United States. To develop an eco-friendly sustainable management strategy for this pest, in this study, we evaluated the efficacy and compatibility of two commercially available biological control agents, predatory beetle Delphastus catalinae (Dc) and entomopathogenic fungi Cordyceps fumosorosea (Cfr) under laboratory conditions. Results showed both Cfr, treatments (alone or in combination with Dc) were efficacious, and Cfr did not negatively impact the beetle longevity. This study’s outcome is important not only for organic horticultural growers, but also for conventional growers as it offers a low-risk alternative tool for solanum whitefly integrated pest management in Florida and other affected regions. Abstract Aleurothrixus (formerly known as Aleurotrachelus) trachoides Back (Hemiptera: Aleyrodidae), commonly known as pepper or solanum whitefly, is a new emerging whitefly pest of several horticultural crops in the United States. During the preliminary survey for pepper whitefly infestation in Florida, a whitefly-specific predatory beetle Delphastus catalinae Horn (Coleoptera: Coccinellidae) was observed associated with this pest in the natural ecosystem. The current study was undertaken to determine the efficacy of this naturally occurring predator, D. catalinae, and an entomopathogenic fungus, Cordyceps (formerly known as Isaria) fumosorosea (Cfr) (Wize) (Hypocreales: Cordycipitaceae), alone or in combination, under controlled laboratory conditions. Whitefly mortality for Cfr (88%), beetle (100%) and Cfr + beetle (100%) treatments were similar and significantly higher compared to control. In the combination treatment, Cfr did not impact D. catalinae longevity and daily food intake, indicating a neutral interaction between the two treatments. Results showed that both of these natural biological control agents could potentially offer an effective alternative in the battle against invasive whiteflies such as pepper whitefly in U.S. horticulture production, either as a stand-alone strategy or in an integrated approach. Although these findings are promising, compatibility of the two treatments needs to be evaluated further under greenhouse and semi-field conditions before recommending to commercial growers.

The ladybird Cheilomenes sexmaculata is reported for the first time from Colombia. Previously, this species wasrecorded in Asia and Australia, then introduced to South America and reported from countries such as Chile, Ecuador,Perú, and Venezuela, where it is used for biological control.

Satellite DNAs are the major repetitive DNA components in eukaryotic genomes. Although satellite DNA has long been called "parasite DNA" there is substantial evidence that it could be associated with some functions of chromosome biology. Ladybird beetles (Coccinellidae) are one of the largest and most important groups of beetles. Many ladybirds are of economic interest as biological control agents because they eat some agricultural pests such as aphids and scale insects. However, other species are phytophagous and can damage crops. Despite the ecological importance of the latter group there are no studies on their satellite DNA. A satellite DNA family was isolated and characterized in the ladybird Henosepilachna argus. This satellite DNA is organized in tandem repeats of 658 bp and is A + T rich (67.3%). The recorded high sequence conservation of the monomers together with the detection of putative gene conversion processes indicate concerted evolution. Reverse transcription polymerase chain reaction (RT-PCR) revealed that this satellite DNA is transcribed and in situ hybridization its location in the subtelomeric regions of all chromosomes except the long arm of the X chromosome. The presence of this satellite DNA in other species of the genus Henosepilachna and Epilachna was also tested using PCR. The results indicate that this satellite DNA sequence is so far specific to H. argus.

Differences in the time of occurrence of both sexes of adult ladybirds, which reflect differences in the longevity and reproductive strategies of the sexes, is important from the perspective of applied entomology, including biological control, as well as basic entomology. Nevertheless, there is little field census data on this topic. We investigated the seasonal occurrence of both sexes of Cheilomenes sexmaculata (Fabricius) in Osaka City, central Japan, from 2003 to 2011. Female adults tended to appear earlier than males after hibernation or aestivation. That males never appeared earlier than females during the nine year period of this study indicate that protogyny occurs after hibernation and aestivation in this species.

Hippodamia variegata is one of the most commercialized ladybirds used for the biological control of aphid pest species in many economically important crops. This species is the first Coccinellidae whose satellitome has been studied by applying new sequencing technologies and bioinformatics tools. We found that 47% of the H. variegata genome is composed of repeated sequences. We identified 30 satellite DNA (satDNA) families with a median intragenomic divergence of 5.75% and A+T content between 45.6% and 74.7%. This species shows satDNA families with highly variable sizes although the most common size is 100&ndash, 200 bp. However, we highlight the existence of a satDNA family with a repeat unit of 2 kb, the largest repeat unit described in Coleoptera. PCR amplifications for fluorescence in situ hybridization (FISH) probe generation were performed for the four most abundant satDNA families. FISH with the most abundant satDNA family as a probe shows its pericentromeric location on all chromosomes. This location is coincident with the heterochromatin revealed by C-banding and DAPI staining, also analyzed in this work. Hybridization signals for other satDNA families were located only on certain bivalents and the X chromosome. These satDNAs could be very useful as chromosomal markers due to their reduced location.

The ladybird beetle, Cheilomenes sexmaculata (Fabricius), has been extending its distribution in and around urban areas at higher latitudes in Japan over the past 100 years. Between 2003 and 2011, we investigated the seasonal occurrence, aphid prey and population dynamics of this species in an urban park in Osaka City, central Japan. We found that C. sexmaculata completes three generations a year in Osaka. Overwintered adults emerge in March or April and produce two (or rarely one) generations by summer. Second-generation adults aestivate and subsequently produce another generation in autumn. This species feeds on five species of aphids that infest planted shrubs and alien weeds throughout the vegetative season. We analyzed the association between accumulated temperature and when overwintered adults first emerged. In addition, we determined the association between their time of emergence, peak abundance and last occurrence, and meteorological conditions. There was no association between the sum of effective temperatures and termination of adult overwintering. There was, however, an association between first occurrence and peak abundance, and climatic conditions, such as warm temperatures or low humidity. This species maintained a stable population in an urban park by becoming quiescent when climatic conditions were unfavourable or prey was scarce. We discussed these findings in relation to urban environmental factors, such as climate, food conditions and vegetation.

Arthropods exhibit a large variety of sex determination systems both at the chromosomal and molecular level. Male heterogamety, female heterogamety, and haplodiploidy occur frequently, but partially different genes are involved. Endosymbionts, such as Wolbachia, Cardinium,Rickettsia, and Spiroplasma, can manipulate host reproduction and sex determination. Four major reproductive manipulation types are distinguished: cytoplasmic incompatibility, thelytokous parthenogenesis, male killing, and feminization. In this review, the effects of these manipulation types and how they interfere with arthropod sex determination in terms of host developmental timing, alteration of sex determination, and modification of sexual differentiation pathways are summarized. Transitions between different manipulation types occur frequently which suggests that they are based on similar molecular processes. It is also discussed how mechanisms of reproductive manipulation and host sex determination can be informative on each other, with a special focus on haplodiploidy. Future directions on how the study of endosymbiotic manipulation of host reproduction can be key to further studies of arthropod sex determination are shown.

Transmission of plant pathogens through insect vectors is a complex biological process involving interactions between the host plants, insects, and pathogens. Simultaneous impact of the insect damage and pathogenic bacteria in infected host plants induce volatiles that modify not only the behavior of its insect vector but also of their natural enemies, such as parasitoid wasps. Therefore, it is essential to understand how insects such as the predator ladybird beetle responds to volatiles emitted from a host plant and how the disease transmission alters the interactions between predators, vector, pathogens, and plants. In this study, we investigated the response of Propylaea japonica to volatiles from citrus plants damaged by Diaphorina citri and Candidatus Liberibacter asiaticus through olfactometer bioassays. Synthetic chemical blends were also used to determine the active compounds in the plant volatile. The results showed that volatiles emitted by healthy plants attracted more P. japonica than other treatments, due to the presence of high quantities of D-limonene and beta-ocimene, and the lack of methyl salicylate. When using synthetic chemicals in the olfactory tests, we found that D-limonene attracted P. japonica while methyl salicylate repelled the predator. However, beta-ocimene attracted the insects at lower concentrations but repelled them at higher concentrations. These results indicate that P. japonica could not efficiently search for its host by using volatile cues emitted from psyllids- and Las bacteria-infected citrus plants.

A new species of Scymnobius Casey (Coleoptera, Coccinellidae, Scymnini) from Pernambuco, Brazil. Scymnobius pernambucensis sp. nov. from Pernambuco, Brazil, is described and illustrated. This is the third species of this genus recorded from Brazil.

The effect of temperature on the functional response of female adults of the two-spot ladybird, Adalia bipunctata L. (Coleoptera: Coccinellidae) was examined in petri dish arenas containing sweet pepper leaves infested with different densities of the green peach aphid, Myzus persicae (Sulzer) (Hemiptera: Aphididae). The predator showed a type II functional response at three tested temperatures ranging from 19A degrees C to 27A degrees C. The theoretical maximum number of prey captured by the predator increased with temperature. Based on the random predator equation, the estimated attack rates ranged from 0.13 h(-1) at 19A degrees C to 0.35 h(-1) at 27A degrees C on a leaf area of 20-25 cm(2). There was no significant difference between the attack rates of the predator at 23A degrees C and 27A degrees C. Handling time significantly decreased as temperature increased from 19A degrees C (0.39 h) to 27A degrees C (0.24 h). This study shows that A. bipunctata displays high predation rates on M. persicae for a wide range of temperatures, indicating its potential for augmentative releases against this aphid pest. The limitations of the predictions generated by functional response experiments are discussed.

While the effect of several factors such as predator and prey size, morphology and developmental stage on intraguild predation (IGP) is widely investigated, little is known about the influence of diet on the occurrence and outcome of IGP. In the present study, the effect of the diet experienced during larval development on IGP between the ladybird Harmonia axyridis and the syrphid Episyrphus balteatus is investigated. Four diets were tested for H. axyridis: eggs of the Mediterranean flour moth Ephestia kuehniella, pea aphids, Acyrthosiphon pisum, in an ad libitum amount, pea aphids in a limited amount, and honey bee pollen. For E. balteatus only the two aphid diets were tested. First, experiments were performed to determine the quality of the various diets for development of both predators. Second, IGP experiments between H. axyridis and E. balteatus were performed both in Petri dishes and on potted pepper plants. The diet of both species influenced the incidence of IGP between H. axyridis and E. balteatus both in Petri dishes and on potted plants. In general, smaller larvae of H. axyridis (those fed on poor or restricted diet) fed more on hoverflies than large (well-nourished) ladybird larvae. Further, poorly nourished (smaller) larvae of E. balteatus were more susceptible to predation than well-fed (larger) hoverfly larvae. The observed effects were not only due to the lower fitness of larvae of both predators reared on an inferior quality diet but also to changes in predator behaviour. The results from this study show that IGP interactions are influenced by a multitude of factors, including feeding history of the organisms involved, and emphasize the importance of taking these factors into account in order to fully understand the ecological relevance of IGP.

The objective of this work was to establish and test the induced-feeding bioassay in order to detect Bacillus thuringiensis insecticidal activity against Epilachna paenulata (Coleoptera: Coccinellidae). Larvae were induced to swallow high concentrations of spore-crystal suspensions of seven exotic and 30 Argentine B. thuringiensis strains. The great majority of strains showed no toxicity to E. paenulata larvae, and observed mortality was lower than 30%. Induced-feeding bioassay is feasible, and should be used for prospecting strains that produce right combinations of Cry proteins needed to an efficient pest control.

The aphid predator Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) is an invasive alien species in Europe and North America with negative effects on non-target species (including a decline of native ladybird populations), as well as fruit production, and human health. It is, therefore, important to find out which natural enemies could be used to reduce their numbers. Knowledge of H. axyridis’ natural enemies is summarised and data collected from the Netherlands over the past ten years are presented. Beetles were sampled from winter aggregations and from spring through to autumn with illuminated screens at night. Natural enemies were not found in samples of H. axyridis from 2003–2007. From 2008 onward H. axyridis adults were infested by: Hesperomyces virescens Thaxt. fungi (summer and winter), Parasitylenchus bifurcatus Poinar and Steenberg nematodes (winter), Coccipolipus hippodamiae (McDaniel and Morrill) mites (winter), and Dinocampus coccinellae (Schrank) parasitoids (summer and winter). Our results indicate that these natural enemies are starting to use H. axyridis as a host, but are as yet not sufficiently abundant to control the population.

Ethanol extracts obtained from the leaf of the thorn apple (Datura stramonium L.) (Solanaceae) were examined for lethal and repellent properties against adults of the European red mite Panonychus ulmi (Koch) (Acari: Tetranychidae) and its predator Stethorus gilvifrons (Muls.) (Col.: Coccinellidae) under laboratory conditions. The Petri leaf disc-spray tower method was used. The results showed that D. stramonium leaf extracts were lethal to both the mite and its predator. Furthermore, an increase in the dose of leaf extract caused a significant increase in the death rates of both P. ulmi and S. gilvifrons adults. Probit analysis and parameter estimation showed that the LC50 values of leaf extract were 7097.5 and 1853.9mg/l at 24h residual activity for spider mite and ladybird adults, respectively. The mite and ladybird mortality rates associated with the extract were higher at 48h than at 24h, with LC50 values of 2013 and 1030mg/l, respectively. However, the LC90 values of the extract were lower for ladybirds than for spider mites at both 24 and 48h. According to Pearson's chi-squared test, both spider mite and ladybird adults showed strong aversions to apple leaf surfaces sprayed with sub-lethal doses of the extract (from 13.72 to 1058.0mg/l). Moreover, an odour test using a two-choice olfactometer demonstrated that ladybird adults prefer fresh air to the extract odour. These results suggest that D. stramonium extract can be used effectively as a safer phytopesticidal product for the management of P. ulmi in both organic and inorganic apple production, but it is not suitable for use in integrated pest management strategies due to its toxic and repellent effects against S. gilvifrons.

The ladybird beetles Propylea quatuordecimpunctata and P. japonica have largely overlapping distributions in northern Japan, and in the laboratory produce fertile hybrids. In this study, we surveyed the distribution and morphological differentiation of these species and the hybrids in natural populations, with a focus on western Hokkaido, northern Japan. Phenotypic analyses were conducted for 987 individuals collected at 90 localities. In addition, the nuclear internal transcribed spacer-II (ITS2) region (549 bp) and part of the mitochondrial cytochrome c oxidase subunit I (COI) gene (700 bp) were sequenced for 620 individuals from 53 localities. Analyses of both phenotypic and genotypic features discriminated two distinct entities assignable to P. quatuordecimpunctata and P. japonica. However, individuals with intermediate phenotypes and/or genotypes also occurred extensively, indicating natural hybridization. Putative P. quatuordecimpunctata individuals were collected across a wide range of altitudes (30-600 m), whereas those of P. japonica were found mostly lower than 300 m alt. In addition, P. quatuordecimpunctata was dominant in semi-open habitats shaded by canopy foliage, whereas P. japonica was frequent in more open habitats. The perceived altitudinal difference in the distributions may thus in part be a consequence of this different habitat preference, as open habitats are more common at lower altitudes in the study area.

The ladybird beetle Stethorus gilvifrons (Muls.) does not provide sufficient mite control in many of the apple orchards in Bursa, despite being an effective and voracious predator of the European red mite, Panonychus ulmi (Koch). This is due in large part to a heavy dependency on organophosphate (OP) and synthetic pyrethroid (SP) insecticides, which were applied to target key pests. To help understand the effects of the insecticides on the ladybird beetle and its main prey, P. ulmi, this study aims to determine their susceptibilities to the residues of parathion-methyl and bifenthrin and detoxifying enzyme activities, such as carboxylesterase (CarE) and glutathione S-transferase (GST). In addition, the insensitivity of acetylcholineesterase (AChE) to the inhibitors, paraoxon and primicarb, is also determined. There are differences in susceptibility to parathion-methyl, in CarE activity and AChE insensitivity, among the populations of predator ladybird and the spider mite. Although GST might play an important role in detoxifying OPs, there is no relationship between susceptibility to parathion-methyl and bifenthrin and GST activity in both the ladybird and the spider mite. On the other hand, two S. gilvifrons populations were surprisingly tolerant to two insecticides. The disparity in susceptibility to parathion-methyl among the predator populations was due to the responsibility of multiple biochemical resistance mechanisms, including elevated CarE activity and target-site insensitivity. Furthermore, the tolerance of the two populations of ladybirds to bifenthrin was similar to that of two P. ulmi populations collected from the same orchards, which indicates a coadaptation to SPs. Therefore, enhanced detoxification by increased activity of CarE is largely responsible for the observed tolerance to bifenthrin in S. gilvifrons populations. Consequently, these results demonstrate that the ladybird species tend to develop resistance to insecticides in the field.

The incidence of sibling cannibalism in clutches of Harmonia axyridis infected by the male-killing Spiroplasma was com- pared with that in uninfected clutches, and the way in which fitness compensation was realized by sibling cannibalism was investi- gated. Primarily the rate of sibling cannibalism was determined by the hatching rate as all unviable eggs were consumed both in infected and uninfected clutches. Per capita consumption of roughly 0.1 individuals was estimated for uninfected clutches in the pre- sent study, as compared to 0.3 individuals in previous studies. The per capita consumption in infected clutches was 1.4, showing that the male-killing behavior of the Spiroplasma provides an approximately 4-14 fold increase in the chance of sibling cannibalism. Both in infected and uninfected clutches, the median of the starting time of sibling cannibalism was soon after that of the completion of hatching. Larvae started to disperse from their egg clutches only about 7.5-8.5 h after the initiation of hatching. These time sequences indicate that sibling cannibalism occurs at the earlier stage of the hatching process and the density of aphids in the area has little or no influence on incidence of the sibling cannibalism. During the maintenance of the infected line for five generations, exclusively females were produced and the median of the hatching rate was 0.395, suggesting a very high rate of vertical transmis- sion of the Spiroplasma infection. Although the measurement of other parameters, such as the effect of Spiroplasma infection on fecundity or longevity, are necessary, the high compensation rate shown in the present study, together with the very high vertical transmission, may explain the prevalence of the present male-killing agent in the local populations of the ladybird beetle.