Reed M. Johnson

A deficit of detoxification enzymes: pesticide sensitivity and environmental response in the honeybee

The honeybee genome has substantially fewer protein coding genes (≈ 11 000 genes) than Drosophila melanogaster (≈ 13 500) and Anopheles gambiae (≈ 14 000). Some of the most marked differences occur in three superfamilies encoding xenobiotic detoxifying enzymes. Specifically there are only about half as many glutathione‐S‐transferases (GSTs), cytochrome P450 monooxygenases (P450s) and carboxyl/cholinesterases (CCEs) in the honeybee. This includes 10‐fold or greater shortfalls in the numbers of Delta and Epsilon GSTs and CYP4 P450s, members of which clades have been recurrently associated with insecticide resistance in other species. These shortfalls may contribute to the sensitivity of the honeybee to insecticides. On the other hand there are some recent radiations in CYP6, CYP9 and certain CCE clades in A. mellifera that could be associated with the evolution of the hormonal and chemosensory processes underpinning its highly organized eusociality.

Genome sequences of the human body louse and its primary endosymbiont provide insights into the permanent parasitic lifestyle

As an obligatory parasite of humans, the body louse (Pediculus humanus humanus) is an important vector for human diseases, including epidemic typhus, relapsing fever, and trench fever. Here, we present genome sequences of the body louse and its primary bacterial endosymbiont Candidatus Riesia pediculicola. The body louse has the smallest known insect genome, spanning 108 Mb. Despite its status as an obligate parasite, it retains a remarkably complete basal insect repertoire of 10,773 protein-coding genes and 57 microRNAs. Representing hemimetabolous insects, the genome of the body louse thus provides a reference for studies of holometabolous insects. Compared with other insect genomes, the body louse genome contains significantly fewer genes associated with environmental sensing and response, including odorant and gustatory receptors and detoxifying enzymes. The unique architecture of the 18 minicircular mitochondrial chromosomes of the body louse may be linked to the loss of the gene encoding the mitochondrial single-stranded DNA binding protein. The genome of the obligatory louse endosymbiont Candidatus Riesia pediculicola encodes less than 600 genes on a short, linear chromosome and a circular plasmid. The plasmid harbors a unique arrangement of genes required for the synthesis of pantothenate, an essential vitamin deficient in the louse diet. The human body louse, its primary endosymbiont, and the bacterial pathogens that it vectors all possess genomes reduced in size compared with their free-living close relatives. Thus, the body louse genome project offers unique information and tools to use in advancing understanding of coevolution among vectors, symbionts, and pathogens.

Pesticides and honey bee toxicity - USA*

Until 1985 discussions of pesticides and honey bee toxicity in the USA were focused on pesticides applied to crops and the unintentional exposure of foraging bees to them. The recent introduction of arthropod pests of honey bees, Acarapis woodi (1984), Varroa destructor (1987), and Aethina tumida (1997), to the USA have resulted in the intentional introduction of pesticides into beehives to suppress these pests. Both the unintentional and the intentional exposure of honey bees to pesticides have resulted in residues in hive products, especially beeswax. This review examines pesticides applied to crops, pesticides used in apiculture and pesticide residues in hive products. We discuss the role that pesticides and their residues in hive products may play in colony collapse disorder and other colony problems. Although no single pesticide has been shown to cause colony collapse disorder, the additive and synergistic effects of multiple pesticide exposures may contribute to declining honey bee health.ZusammenfassungNeuere systemisch wirkende Pestizide, einschließlich der Neonikotinoide (z. B. Imidacloprid) und Phenylpyrazole (z. B. Fipronil) finden in den USA verbreitete Anwendung im Pflanzenschutz. Das Gefährdungspotenzial von Bienen durch diese Präparate unterscheidet sich von dem traditioneller Pestizidanwendungen, bei denen die hauptsächliche Sorge der akuten Giftigkeit galt. Im Hinblick auf die Verordnungen zu Pestiziden in den USA wurden die Folgen von chronischer und sublethaler Belastung durch systemische Mittel bisher nicht umfassend in Betracht gezogen, obwohl die Sachlage, was diese Präparate betrifft, gegenwärtig von der Umweltbehörde (EPA) begutachtet wird. Zahlreiche in den USA angebaute Pflanzen wurden genetisch verändert, um entweder insektizid wirkende Bt Toxine oder Herbizidresistenz zu exprimieren. Insektizid wirkende Bt Toxine scheinen jedoch spezifisch toxisch für Ertragsschädlinge zu sein und können daher den Bienen nützen, indem sie die Anwendung traditioneller Pestizide reduzieren.Bis zur Einführung von arthropoden Bienenschädlingen in die USA in der Mitte der achtziger Jahre wurden Bienen den verschiedenen Pestiziden nur unbeabsichtigt ausgesetzt, während sie auf gespritzten Pflanzen sammelten. Die Notwendigkeit, Bienenschädlinge, besonders die Varroamilbe (Varroa destructor), zu bekämpfen, erfordert seitdem jedoch oft eine absichtliche Anwendung von Pestiziden in Bienenvölkern. Tau-Fluvalinat und Coumaphos, jeweils in Streifenform angewendet, sind in den USA immer noch für die Anwendung in Bienenvölkern zugelassen, obwohl die Wirksamkeit dieser Substanzen gegen Varroamilben durch die Entwicklung von Resistenzen vermindert wurde. Ein neues Varroazid, Fenpyroximate, wurde in einigen Staaten zur Anwendung zugelassen. Essentielle Öle, einschließlich Thymol und Menthol, sind ebenso wie Ameisensäure zur Anwendung in der Verdampfung zugelassen. Oxalsäure ist nur in Kanada, jedoch nicht in den USA zugelassen.Über 150 verschiedene Pestizide wurden in Proben aus Bienenständen in den USA gefunden. Von Imkern eingesetzte Pestizide werden tendenziell öfter im Wachs der Völker nachgewiesen, von wo aus Pollen, Bienenbrot und Honig damit kontaminiert werden. Auf der anderen Seite werden Pestizide, vor allem Fungizide, die nicht in Bienenvölkern eingesetzt werden, tendenziell am häufigsten in Pollen nachgewiesen und kontaminieren das Wachs nur dann, wenn sie eingelagert werden. Da Honigbienen den sublethalen Konzentrationen zahlreicher Pestizide gleichzeitig ausgesetzt sind, wird zusätzliche Forschung zur Aufklärung synergistischer Effekte bei chronischer sublethaler Belastung mit mehreren Pesitziden benötigt.

Draft genome of the red harvester ant Pogonomyrmex barbatus

We report the draft genome sequence of the red harvester ant, Pogonomyrmex barbatus. The genome was sequenced using 454 pyrosequencing, and the current assembly and annotation were completed in less than 1 y. Analyses of conserved gene groups (more than 1,200 manually annotated genes to date) suggest a high-quality assembly and annotation comparable to recently sequenced insect genomes using Sanger sequencing. The red harvester ant is a model for studying reproductive division of labor, phenotypic plasticity, and sociogenomics. Although the genome of P. barbatus is similar to other sequenced hymenopterans (Apis mellifera and Nasonia vitripennis) in GC content and compositional organization, and possesses a complete CpG methylation toolkit, its predicted genomic CpG content differs markedly from the other hymenopterans. Gene networks involved in generating key differences between the queen and worker castes (e.g., wings and ovaries) show signatures of increased methylation and suggest that ants and bees may have independently co-opted the same gene regulatory mechanisms for reproductive division of labor. Gene family expansions (e.g., 344 functional odorant receptors) and pseudogene accumulation in chemoreception and P450 genes compared with A. mellifera and N. vitripennis are consistent with major life-history changes during the adaptive radiation of Pogonomyrmex spp., perhaps in parallel with the development of the North American deserts.

Draft genome of the globally widespread and invasive Argentine ant (Linepithema humile)

Ants are some of the most abundant and familiar animals on Earth, and they play vital roles in most terrestrial ecosystems. Although all ants are eusocial, and display a variety of complex and fascinating behaviors, few genomic resources exist for them. Here, we report the draft genome sequence of a particularly widespread and well-studied species, the invasive Argentine ant (Linepithema humile), which was accomplished using a combination of 454 (Roche) and Illumina sequencing and community-based funding rather than federal grant support. Manual annotation of >1,000 genes from a variety of different gene families and functional classes reveals unique features of the Argentine ants biology, as well as similarities to Apis mellifera and Nasonia vitripennis. Distinctive features of the Argentine ant genome include remarkable expansions of gustatory (116 genes) and odorant receptors (367 genes), an abundance of cytochrome P450 genes (>110), lineage-specific expansions of yellow/major royal jelly proteins and desaturases, and complete CpG DNA methylation and RNAi toolkits. The Argentine ant genome contains fewer immune genes than Drosophila and Tribolium, which may reflect the prominent role played by behavioral and chemical suppression of pathogens. Analysis of the ratio of observed to expected CpG nucleotides for genes in the reproductive development and apoptosis pathways suggests higher levels of methylation than in the genome overall. The resources provided by this genome sequence will offer an abundance of tools for researchers seeking to illuminate the fascinating biology of this emerging model organism.

Changes in transcript abundance relating to colony collapse disorder in honey bees (Apis mellifera)

Colony collapse disorder (CCD) is a mysterious disappearance of honey bees that has beset beekeepers in the United States since late 2006. Pathogens and other environmental stresses, including pesticides, have been linked to CCD, but a causal relationship has not yet been demonstrated. Because the gut acts as a primary interface between the honey bee and its environment as a site of entry for pathogens and toxins, we used whole-genome microarrays to compare gene expression between guts of bees from CCD colonies originating on both the east and west coasts of the United States and guts of bees from healthy colonies sampled before the emergence of CCD. Considerable variation in gene expression was associated with the geographical origin of bees, but a consensus list of 65 transcripts was identified as potential markers for CCD status. Overall, elevated expression of pesticide response genes was not observed. Genes involved in immune response showed no clear trend in expression pattern despite the increased prevalence of viruses and other pathogens in CCD colonies. Microarray analysis revealed unusual ribosomal RNA fragments that were conspicuously more abundant in the guts of CCD bees. The presence of these fragments may be a possible consequence of picorna-like viral infection, including deformed wing virus and Israeli acute paralysis virus, and may be related to arrested translation. Ribosomal fragment abundance and presence of multiple viruses may prove to be useful diagnostic markers for colonies afflicted with CCD.

Acaricide, Fungicide and Drug Interactions in Honey Bees (Apis mellifera)

Background Chemical analysis shows that honey bees (Apis mellifera) and hive products contain many pesticides derived from various sources. The most abundant pesticides are acaricides applied by beekeepers to control Varroa destructor. Beekeepers also apply antimicrobial drugs to control bacterial and microsporidial diseases. Fungicides may enter the hive when applied to nearby flowering crops. Acaricides, antimicrobial drugs and fungicides are not highly toxic to bees alone, but in combination there is potential for heightened toxicity due to interactive effects. Methodology/Principal Findings Laboratory bioassays based on mortality rates in adult worker bees demonstrated interactive effects among acaricides, as well as between acaricides and antimicrobial drugs and between acaricides and fungicides. Toxicity of the acaricide tau-fluvalinate increased in combination with other acaricides and most other compounds tested (15 of 17) while amitraz toxicity was mostly unchanged (1 of 15). The sterol biosynthesis inhibiting (SBI) fungicide prochloraz elevated the toxicity of the acaricides tau-fluvalinate, coumaphos and fenpyroximate, likely through inhibition of detoxicative cytochrome P450 monooxygenase activity. Four other SBI fungicides increased the toxicity of tau-fluvalinate in a dose-dependent manner, although possible evidence of P450 induction was observed at the lowest fungicide doses. Non-transitive interactions between some acaricides were observed. Sublethal amitraz pre-treatment increased the toxicity of the three P450-detoxified acaricides, but amitraz toxicity was not changed by sublethal treatment with the same three acaricides. A two-fold change in the toxicity of tau-fluvalinate was observed between years, suggesting a possible change in the genetic composition of the bees tested. Conclusions/Significance Interactions with acaricides in honey bees are similar to drug interactions in other animals in that P450-mediated detoxication appears to play an important role. Evidence of non-transivity, year-to-year variation and induction of detoxication enzymes indicates that pesticide interactions in bees may be as complex as drug interactions in mammals.

Synergistic interactions between in-hive miticides in Apis mellifera

ABSTRACT The varroa mite, Varroa destructor Anderson & Trueman, is a devastating pest of honey bees, Apis mellifera L., that has been primarily controlled over the last 15 yr with two in-hive miticides: the organophosphate coumaphos (Checkmite+), and the pyrethroid tau-fluvalinate (Apistan). Both coumaphos and tau-fluvalinate are lipophilic compounds that are absorbed by the wax component of the hive, where they are stable and have the potential to build up over repeated treatments such that bees could be exposed to both compounds simultaneously. Although these compounds were chosen as in-hive miticides due to their low toxicity to honey bees, that low toxicity depends, at least in part, on rapid detoxification mediated by cytochrome P450 monooxygenase enzymes (P450s). In this laboratory study, we observed a large increase in the toxicity of tau-fluvalinate to 3-d-old bees that had been treated previously with coumaphos, and a moderate increase in the toxicity of coumpahos in bees treated previously with tau-fluvalinate. The observed synergism may result from competition between miticides for access to detoxicative P450s. These results suggest that honey bee mortality may occur with the application of otherwise sublethal doses of miticide when tau-fluvalinate and coumaphos are simultaneously present in the hive.

Genomic signatures of evolutionary transitions from solitary to group living

For bees, many roads lead to social harmony Eusociality, where workers sacrifice their reproductive rights to support the colony, has evolved repeatedly and represents the most evolved form of social evolution in insects. Kapheim et al. looked across the genomes of 10 bee species with varying degrees of sociality to determine the underlying genomic contributions. No one genomic path led to eusociality, but similarities across genomes were seen in features such as increases in gene regulation and methylation. It also seems that selection pressures relaxed after the emergence of complex sociality. Science, this issue p. 1139 Social evolution in bees has followed diverse genomic paths but shares genomic patterns. The evolution of eusociality is one of the major transitions in evolution, but the underlying genomic changes are unknown. We compared the genomes of 10 bee species that vary in social complexity, representing multiple independent transitions in social evolution, and report three major findings. First, many important genes show evidence of neutral evolution as a consequence of relaxed selection with increasing social complexity. Second, there is no single road map to eusociality; independent evolutionary transitions in sociality have independent genetic underpinnings. Third, though clearly independent in detail, these transitions do have similar general features, including an increase in constrained protein evolution accompanied by increases in the potential for gene regulation and decreases in diversity and abundance of transposable elements. Eusociality may arise through different mechanisms each time, but would likely always involve an increase in the complexity of gene networks.

Metabolic enzymes associated with xenobiotic and chemosensory responses in Nasonia vitripennis.

The numbers of glutathione S‐transferase, cytochrome P450 and esterase genes in the genome of the hymenopteran parasitoid Nasonia vitripennis are about twice those found in the genome of another hymenopteran, the honeybee Apis mellifera. Some of the difference is associated with clades of these families implicated in xenobiotic resistance in other insects and some is in clades implicated in hormone and pheromone metabolism. The data support the hypothesis that the eusocial behaviour of the honeybee and the concomitant homeostasis of the nest environment may obviate the need for as many gene/enzyme systems associated with xenobiotic metabolism as are found in other species, including N. vitripennis, that are thought to encounter a wider range of potentially toxic xenobiotics in their diet and habitat.