Quantitative assessment of plant-arthropod interactions in forest canopies: A plot-based approach.

Research on canopy arthropods has progressed from species inventories to the study of their interactions and networks, enhancing our understanding of how hyper-diverse communities are maintained. Previous studies often focused on sampling individual tree species, individual trees or their parts. We argue that such selective sampling is not ideal when analyzing interaction network structure, and may lead to erroneous conclusions. We developed practical and reproducible sampling guidelines for the plot-based analysis of arthropod interaction networks in forest canopies. Our sampling protocol focused on insect herbivores (leaf-chewing insect larvae, miners and gallers) and non-flying invertebrate predators (spiders and ants). We quantitatively sampled the focal arthropods from felled trees, or from trees accessed by canopy cranes or cherry pickers in 53 0.1 ha forest plots in five biogeographic regions, comprising 6,280 trees in total. All three methods required a similar sampling effort and provided good foliage accessibility. Furthermore, we compared interaction networks derived from plot-based data to interaction networks derived from simulated non-plot-based data focusing either on common tree species or a representative selection of tree families. All types of non-plot-based data showed highly biased network structure towards higher connectance, higher web asymmetry, and higher nestedness temperature when compared with plot-based data. Furthermore, some types of non-plot-based data showed biased diversity of the associated herbivore species and specificity of their interactions. Plot-based sampling thus appears to be the most rigorous approach for reconstructing realistic, quantitative plant-arthropod interaction networks that are comparable across sites and regions. Studies of plant interactions have greatly benefited from a plot-based approach and we argue that studies of arthropod interactions would benefit in the same way. We conclude that plot-based studies on canopy arthropods would yield important insights into the processes of interaction network assembly and dynamics, which could be maximised via a coordinated network of plot-based study sites.

Muscle mass drives cost in sexually selected arthropod weapons.

Sexually selected weapons often function as honest signals of fighting ability. If poor-quality individuals produce high-quality weapons, then receivers should focus on other, more reliable signals. Cost is one way to maintain signal integrity. The costs of weapons tend to increase with relative weapon size, and thereby restrict large weapons to high-quality individuals who can produce and maintain them. Weapon cost, however, appears to be unpredictably variable both within and across taxa, and the mechanisms underlying this variation remain unclear. We suggest variation in weapon cost may result from variation in weapon composition-specifically, differences in the amount of muscle mass directly associated with the weapon. We test this idea by measuring the metabolic cost of sexually selected weapons in seven arthropod species and relating these measures to weapon muscle mass. We show that individuals with relatively large weapon muscles have disproportionately high resting metabolic rates and provide evidence that this trend is driven by weapon muscle mass. Overall, our results suggest that variation in weapon cost can be partially explained by variation in weapon morphology and that the integrity of weapon signals may be maintained by increased metabolic cost in species with relatively high weapon muscle mass.

Ecotoxicological assessment of Fluazuron: effects on Folsomia candida and Eisenia andrei.

The cattle production in Brazil has increased considerably in the last years, mainly due to the control of parasite infestation of the animals, which cause loss of productivity to the sector. Fluazuron is an active ingredient (a.i.) of the benzoylurea class used to control ticks in cattle. As this a.i. has been found unchanged in animal feces, which may present a risk to edaphic organisms, this study aimed to assess the effects of fluazuron on survival, reproduction, and behavior of the soil invertebrates Folsomia candida and Eisenia andrei, through ecotoxicological assays. We carried out bioassays in a tropical artificial soil (TAS) spiked with increasing doses of the insecticide. Earthworm mortality was found only at the highest tested fluazuron concentration (LOEC = 160 mg a.i. kg-1 dry soil and NOEC = 80 mg kg-1), while the reproduction of F. candida and E. andrei was reduced at lower fluazuron concentrations (EC50 = 4.48 mg kg-1 and EC50 = 20.8 mg kg-1, respectively). Avoidance behavior was detected for both species at lower concentrations than those that caused impacts on reproduction, indicating that the substance may affect the soil habitat function. Since the possible adverse effects of fluazuron on edaphic fauna are still unknown or neglected, this study also warns about the possible harmful effect of veterinary pharmaceutical products on edaphic fauna.

Mixture toxicity assessment of a biocidal product based on reproduction and avoidance behaviour of the collembolan Folsomia candida.

Biocidal products represent mixtures that might be released into the environment at application and continuously during service life. Concentration addition (CA) has been proposed as default model to calculate theoretical mixture toxicity. However, the suitability of CA for chronic toxicity towards soil organisms has so far rarely been evaluated and therefore needs further experimental evidence. The present study investigated the toxicity of a wood preservative product and the individual active substances (tebuconazole and IPBC) therein with the aim to evaluate the compliance with the CA prediction for the product. Folsomia candida was selected as test organism for this purpose using the endpoints reproduction and avoidance behaviour. Both endpoints were increasingly impacted by increasing concentrations of the wood preservative product as well as its active substances tested individually. The chronic effects of the product could be predicted by CA with less than 4-fold deviation, while the assessment for avoidance behaviour indicated a strong underestimation. This underestimation could not be attributed to the one known formulation additive, an organic solvent. Overall, the present study provides some more evidence that CA could be applied as default model for standard endpoints of soil organisms, but warns against using CA for behavioural responses.

Ecological hazard assessment of methyl ethyl ketone using the species sensitivity distribution approach in a soil ecosystem.

Methyl ethyl ketone (MEK) is a common and widely used industrial solvent. However, few studies have investigated its toxicity, or its effects as a contaminant in soil ecosystems. In this study, acute and chronic toxicity data for MEK were generated, and ecological risk based on a species sensitivity distribution was assessed. Seven soil organisms from six taxonomic groups were used for acute toxicity tests and five soil organisms from four taxonomic groups were used for chronic toxicity tests. Acute and chronic soil HC5 (hazardous concentration for 5% of species) values for MEK were estimated as 53.04 and 2.593 mg MEK/kg dry soil, respectively. This is the first study to conduct battery testing for MEK; it specifies hazardous concentrations, warns of the need for accident preparedness, and points to serious potential hazards of MEK at various levels of the soil ecosystem which can translate into greater environmental damage with implications for human health. The specific sensitivity levels determined may serve as a benchmark for establishing soil standards and strategies for ecosystem protection in the face of accidental contamination.

Ecotoxicological assessment of biosolids by microcosms.

Biosolids have been applied as soil amendments to improve and maintain the soil fertility and faster plant growth. In spite of its beneficial use, the potential risks of land disposal should be analyzed, considering potential ecological receptors in soil and water. This work describes the use of an early warning laboratory microcosm system to evaluate the integrated ecotoxicological potential of two biosolids: BIO-1 and BIO-2 (18 and 28 months after landfarming, respectively), from an effluent treatment station in a petrochemical and industrial district. The endpoints related to habitat function were: a) germination, growth and biomass of Phaseolus vulgaris; b) survival, biomass and number of cocoons of Eisenia andrei (Oligochaeta) and; c) reproduction of Folsomia candida (Collembola). The retention function was evaluated by testing the leachates using the tropical cladoceran Latonopsis australis (Cladocera) in a 48-h acute toxicity test, and growth of the aquatic plant Lemna minor in a 7-d chronic test. Tropical artificial soil (TAS) and a natural soil (NS) from the region were used as control soils. Results showed no chronic toxicity of BIO-1 and BIO-2 to the soil organisms tested, but acute toxicity of BIO-1 in the leachate for 50% of L. australis, and chronic toxicity of both biosolid leachates to L. minor (inhibition of growth rate), indicating potential risks to aquatic ecosystems. The results confirmed the ability of this microcosm system as a rapid tool to assess biosolid toxicity over time and its potential for hazardous waste characterization in environmental risk assessment, in a screening phase.

Assessment the short-term effects of wrack removal on supralittoral arthropods using the M-BACI design on Atlantic sandy beaches of Brazil and Spain.

Wrack removal has been adopted indiscriminately, with no previous assessment of the ecological implications for sandy beach ecosystem. This study evaluated, through an M-BACI design, the effect of wrack removal on supralittoral arthropods on Atlantic sandy beaches receiving different types of wrack: mangrove propagules (Brazil), seagrasses and macroalgae (Spain). Impacted plots were contrasted with controls in 8 successive periods before and after experimental wrack removal. After the disturbance, drastic decreases in the densities of the amphipod Platorchestia monodi, coleopterans Cleridae, Nitidulidae and Phaleria testacea (Brazilian beaches) and amphipod Talitrus saltator (Spanish beaches) were detected in the impacted plots. The recovery patterns of arthropods might be related to wrack features (amount, composition, and degradation) combined with density and species-specific strategies (e.g. mobility, feeding preferences) in each Atlantic region. The temporary suppression of wrack and its associated fauna can have potential effects on the wrack-derived process and food-web structure on sandy beaches.

Transportable data from non-target arthropod field studies for the environmental risk assessment of genetically modified maize expressing an insecticidal double-stranded RNA.

As part of an environmental risk assessment, the potential impact of genetically modified (GM) maize MON 87411 on non-target arthropods (NTAs) was evaluated in the field. MON 87411 confers resistance to corn rootworm (CRW; Diabrotica spp.) by expressing an insecticidal double-stranded RNA (dsRNA) transcript and the Cry3Bb1 protein and tolerance to the herbicide glyphosate by producing the CP4 EPSPS protein. Field trials were conducted at 14 sites providing high geographic and environmental diversity within maize production areas from three geographic regions including the U.S., Argentina, and Brazil. MON 87411, the conventional control, and four commercial conventional reference hybrids were evaluated for NTA abundance and damage. Twenty arthropod taxa met minimum abundance criteria for valid statistical analysis. Nine of these taxa occurred in at least two of the three regions and in at least four sites across regions. These nine taxa included: aphid, predatory earwig, lacewing, ladybird beetle, leafhopper, minute pirate bug, parasitic wasp, sap beetle, and spider. In addition to wide regional distribution, these taxa encompass the ecological functions of herbivores, predators and parasitoids in maize agro-ecosystems. Thus, the nine arthropods may serve as representative taxa of maize agro-ecosystems, and thereby support that analysis of relevant data generated in one region can be transportable for the risk assessment of the same or similar GM crop products in another region. Across the 20 taxa analyzed, no statistically significant differences in abundance were detected between MON 87411 and the conventional control for 123 of the 128 individual-site comparisons (96.1%). For the nine widely distributed taxa, no statistically significant differences in abundance were detected between MON 87411 and the conventional control. Furthermore, no statistically significant differences were detected between MON 87411 and the conventional control for 53 out of 56 individual-site comparisons (94.6 %) of NTA pest damage to the crop. In each case where a significant difference was observed in arthropod abundance or damage, the mean value for MON 87411 was within the reference range and/or the difference was not consistently observed across collection methods and/or sites. Thus, the differences were not representative of an adverse effect unfamiliar to maize and/or were not indicative of a consistent plant response associated with the GM traits. Results from this study support a conclusion of no adverse environmental impact of MON 87411 on NTAs compared to conventional maize and demonstrate the utility of relevant transportable data across regions for the ERA of GM crops.

The microbiome of Folsomia candida: an assessment of bacterial diversity in a Wolbachia-containing animal.

The springtail Folsomia candida is an important model organism for soil ecology, ecotoxicology and ecogenomics. The decomposer activities of soil invertebrates like Folsomia depend on their relationship with microbial communities including gut symbionts. In this paper, we apply high-throughput sequencing to provide a detailed characterization of the bacterial community associated with parthenogenetic F. candida. First, we evaluated a method to suppress the amplification of DNA from the endosymbiont Wolbachia, to prevent it from interfering with the identification of less abundant operational taxonomic units (OTUs). The suppression treatment applied was effective against Wolbachia and did not interfere with the detection of the most abundant OTUs (59 OTUs, contributing over 87% of the reads). However, this method did affect the inferred community composition. Significant differences were subsequently observed in the composition of bacterial communities associated with two different strains of F. candida. A total of 832 OTUs were found, of which 45% were only present in one strain and 17% only in the other. Among the 20 most abundant OTUs, 16 were shared between strains. Denaturing gradient gel electrophoresis and clone libraries, although unable to capture the full diversity of the bacterial community, provided results that supported the NGS data.

Characterization of the ecological interactions of Roundup Ready 2 Yield® soybean, MON 89788, for use in ecological risk assessment.

As part of an ecological risk assessment, Roundup Ready 2 Yield® soybean (MON 89788) was compared to a conventional control soybean variety, A3244, for disease and arthropod damage, plant response to abiotic stress and cold, effects on succeeding plant growth (allelopathic effects), plant response to a bacterial symbiont, and effects on the ability of seed to survive and volunteer in a subsequent growing season. Statistically significant differences between MON 89788 and A3244 were considered in the context of the genetic variation known to occur in soybean and were assessed for their potential impact on plant pest (weed) potential and adverse environmental impact. The results of these studies revealed no effects of the genetic modification that would result in increased pest potential or adverse environmental impact of MON 89788 compared with A3244. This paper illustrates how such characterization studies conducted in a range of environments where the crop is grown are used in an ecological risk assessment of the genetically modified (GM) crop. Furthermore, risk assessors and decision makers use this information when deciding whether to approve a GM crop for cultivation in-or grain import into-their country.

Short-term effects of different genetically modified maize varieties on arthropod food web properties: an experimental field assessment.

There is concern that genetically modified (GM) plants may have adverse affects on the arthropod biodiversity comprising agricultural landscapes. The present study report on a two year field experimental test of whether four different genotypic lines, some are novel with no previous field tests, of GM maize hybrids alter the structure of arthropod food webs that they harbour, relative to non-GM maize (control) that is widely used in agriculture. The different GM genotypes produced either Bt toxins, conferred glyphosate tolerance or a combination of the two traits. Quantitative food web analysis, based on short-term assessment assigning a total of 243,896 arthropod individuals collected from the treatments to their positions in food webs, revealed that complex and stable food webs persisted in each maize treatment. Moreover, food web structure remained relatively unchanged by the GM-genotype. The results suggest that at least in short-term period these particular GM maize genotypes will not have adverse effects on arthropod biota of agricultural landscapes.

Potential use of an arthropod database to support the non-target risk assessment and monitoring of transgenic plants.

Worldwide, plants obtained through genetic modification are subject to a risk analysis and regulatory approval before they can enter the market. An area of concern addressed in environmental risk assessments is the potential of genetically modified (GM) plants to adversely affect non-target arthropods and the valued ecosystem services they provide. Environmental risk assessments are conducted case-by-case for each GM plant taking into account the plant species, its trait(s), the receiving environments into which the GM plant is to be released and its intended uses, and the combination of these characteristics. To facilitate the non-target risk assessment of GM plants, information on arthropods found in relevant agro-ecosystems in Europe has been compiled in a publicly available database of bio-ecological information during a project commissioned by the European Food Safety Authority (EFSA). Using different hypothetical GM maize case studies, we demonstrate how the information contained in the database can assist in identifying valued species that may be at risk and in selecting suitable species for laboratory testing, higher-tier studies, as well as post-market environmental monitoring.

A simulation study on effects of exposure to a combination of pesticides used in an orchard and tuber crop on the recovery time of a vulnerable aquatic invertebrate.

The aim of the present study was to assess whether population effects and recovery times increase when a population of a vulnerable aquatic invertebrate is exposed to concentrations of 1 or multiple pesticides. The 2 sets of pesticide combinations tested are typical for orchard and tuber crops in The Netherlands. Exposure concentrations were predicted using the FOCUS step 3 modeling framework and the Dutch drainage ditch scenario. Recovery times were assessed using the MASTEP population model. We simulated the population dynamics and pesticide effects in a Monte Carlo style by using median effective concentration values drawn from an arthropod species sensitivity distribution. In the tuber scenario, exposure to λ-cyhalothrin resulted in long-term effects, whereas exposure to the co-occurring compound fluazinam hardly resulted in (additional) effects. In the orchard scenario, 3 pesticides resulted in large effects just after exposure, but pulse exposures to these compounds did not coincide. The probabilities of effects for the single compounds added up for the combination; in contrast, the recovery times were not higher for the combination compared to those associated with exposure to the individual compounds. The conclusion from the present study's simulations is that exposure to the evaluated pesticide packages may lead to increased mortality probabilities and effect sizes of the combination, but does not lead to longer recovery times for populations with synchronized reproduction than when exposed to the individual compounds.

Artificial light pollution: are shifting spectral signatures changing the balance of species interactions?

Technological developments in municipal lighting are altering the spectral characteristics of artificially lit habitats. Little is yet known of the biological consequences of such changes, although a variety of animal behaviours are dependent on detecting the spectral signature of light reflected from objects. Using previously published wavelengths of peak visual pigment absorbance, we compared how four alternative street lamp technologies affect the visual abilities of 213 species of arachnid, insect, bird, reptile and mammal by producing different wavelength ranges of light to which they are visually sensitive. The proportion of the visually detectable region of the light spectrum emitted by each lamp was compared to provide an indication of how different technologies are likely to facilitate visually guided behaviours such as detecting objects in the environment. Compared to narrow spectrum lamps, broad spectrum technologies enable animals to detect objects that reflect light over more of the spectrum to which they are sensitive and, importantly, create greater disparities in this ability between major taxonomic groups. The introduction of broad spectrum street lamps could therefore alter the balance of species interactions in the artificially lit environment.

Ecotoxicological assessment of metal-polluted urban soils using bioassays with three soil invertebrates.

This study aimed at assessing the quality of urban soils by integrating chemical and ecotoxicological approaches. Soils from five sites in downtown Naples, Italy, were sampled and characterized for physical-chemical properties and total and water-extractable metal concentrations. Bioassays with Eisenia andrei, Enchytraeus crypticus and Folsomia candida were performed to assess toxicity of the soils, using survival, reproduction and growth as the endpoints. Metal bioaccumulation in the animals was also measured. The properties and metal concentrations of the soils strongly differed. Metal bioaccumulation was related with total metal concentrations in soil and was highest in E. crypticus, which was more sensitive than E. andrei and F. candida. Responses of the three species to the investigated soils seemed due to both metal contamination and soil properties.

Seasonal activity of millipedes (Diplopoda)--their economic and medical significance.

The millipede Brachydesmus superus Latzel, Polydesmus inconstans Latzel (Diplopoda: Polydesmida) and Kryphioiulus occultus C. L. Koch (Diplopoda: Julida) were collected from compost in gardens in Lublin, eastern Poland. Collections were made by using pitfall traps between April - September 2009 and 2010. Brachydesmus superus, Polydesmus inconstans and Kryphioiulus occultus play a significant role in composting of plant residues. Cylindroiulus caeruleocinctus Wood and Ommatoiulus sabulosus Linnaeus (Diplopoda: Julida) were collected manually in 2009-2011 in fallows and ruderals of Lublin and Kraków. C. caeruleocinctus and O. sabulosus caused considerable nuisance during mass occurrence and migration in human residences. The sex ratio has been determined for populations of C. caeruleocinctus, total sex ratio average 1:1.46. The number of females prevailed. Millipedes of the temperate climate have two peaks in the spring and autumn pattern of activity of the year. Both common species C. caeruleocinctus and O. sabulosus were analysed bacteriologically. The millipede Cylindroiulus caeruleocinctus transmits Citrobacter freundii, Pantoea agglomerans, Serratia marcescens, Raoultella planticola, Salmonella arizonae. The millipede Ommatoiulus sabulosus transmits Citrobacter freundii, Pantoea agglomerans, Raoultella planticola and Xanthomonas maltophila.

Invasive arthropods.

Many arthropod species have been transported around the globe and successfully invaded new regions. Invasive arthropods can have severe impacts on animal and human health, agriculture and forestry, and the biodiversity of natural habitats as well as those modified by humans. The economic and environmental effects of invasion can be both direct, through feeding and competition, and indirect, such as the transmission of pathogens. In this paper, the authors consider ten examples that illustrate the main mechanisms of introduction, the characteristics that enable species to rapidly expand their ranges and some of the consequences of their arrival.

Reproductive flexibility: genetic variation, genetic costs and long-term evolution in a collembola.

In a variable yet predictable world, organisms may use environmental cues to make adaptive adjustments to their phenotype. Such phenotypic flexibility is expected commonly to evolve in life history traits, which are closely tied to Darwinian fitness. Yet adaptive life history flexibility remains poorly documented. Here we introduce the collembolan Folsomia candida, a soil-dweller, parthenogenetic (all-female) microarthropod, as a model organism to study the phenotypic expression, genetic variation, fitness consequences and long-term evolution of life history flexibility. We demonstrate that collembola have a remarkable adaptive ability for adjusting their reproductive phenotype: when transferred from harsh to good conditions (in terms of food ration and crowding), a mother can fine-tune the number and the size of her eggs from one clutch to the next. The comparative analysis of eleven clonal populations of worldwide origins reveals (i) genetic variation in mean egg size under both good and bad conditions; (ii) no genetic variation in egg size flexibility, consistent with convergent evolution to a common physiological limit; (iii) genetic variation of both mean reproductive investment and reproductive investment flexibility, associated with a reversal of the genetic correlation between egg size and clutch size between environmental conditions ; (iv) a negative genetic correlation between reproductive investment flexibility and adult lifespan. Phylogenetic reconstruction shows that two life history strategies, called HIFLEX and LOFLEX, evolved early in evolutionary history. HIFLEX includes six of our 11 clones, and is characterized by large mean egg size and reproductive investment, high reproductive investment flexibility, and low adult survival. LOFLEX (the other five clones) has small mean egg size and low reproductive investment, low reproductive investment flexibility, and high adult survival. The divergence of HIFLEX and LOFLEX could represent different adaptations to environments differing in mean quality and variability, or indicate that a genetic polymorphism of reproductive investment reaction norms has evolved under a physiological tradeoff between reproductive investment flexibility and adult lifespan.

Avoidance tests in site-specific risk assessment--influence of soil properties on the avoidance response of Collembola and earthworms.

The ability of organisms to avoid contaminated soils can act as an indicator of toxic potential in a particular soil. Based on the escape response of earthworms and Collembola, avoidance tests with these soil organisms have great potential as early screening tools in site-specific assessment. These tests are becoming more common in soil ecotoxicology, because they are ecologically relevant and have a shorter duration time compared with standardized soil toxicity tests. The avoidance response of soil invertebrates, however, can be influenced by the soil properties (e.g., organic matter content and texture) that affect behavior of the test species in the exposure matrix. Such an influence could mask a possible effect of the contaminant. Therefore, the effects of soil properties on performance of test species in the exposure media should be considered during risk assessment of contaminated soils. Avoidance tests with earthworms (Eisenia andrei) and springtails (Folsomia candida) were performed to identify the influence of both organic matter content and texture on the avoidance response of representative soil organisms. Distinct artificial soils were prepared by modifying quantities of the standard artificial soil components described by the Organization for Economic Co-operation and Development to achieve different organic matter and texture classes. Several combinations of each factor were tested. Results showed that both properties influenced the avoidance response of organisms, which avoided soils with low organic matter content and fine texture. Springtails were less sensitive to changes in these soil constituents compared with earthworms, indicating springtails can be used for site-specific assessments of contaminated soils with a wider range of respective soil properties.

9th International Symposium on the Biosafety of Genetically Modified Organisms. Session II: Identifying and defining hazards and potential consequences I: Concepts for problem formulation and non-target risk assessment.

The scientific organizers of the symposium put much emphasis on the identification and definition of hazard and the potential consequences thereof and three full sessions with a total of 13 presentations encompassing a wide range of related themes were planned for this topic. Unfortunately, one talk had to be cancelled because of illness of the speaker (BM Khadi, India). Some presentations covered conceptual approaches for environmental risk assessment (ERA) of GM plants (problem formulation in the risk assessment framework, familiarity approach, tiered and methodological frameworks, non-target risk assessment) and the use of models in assessing invasiveness and weediness of GM plants. Other presentations highlighted the lessons learned for future ERA from case studies and commercialized GM crops, and from monitoring of unintended releases to the environment. When the moderators of the three sessions came together after the presentations to align their summaries, there was an obvious need to restructure the 12 presentations in a way that allowed for a consistent summarizing discussion. The following new organization of the 12 talks was chosen: (1) Concepts for problem formulation and non-target risk assessment, (2) Modeling as a tool for predicting invasiveness of GM plants, (3) Case-studies of ERA of large-scale release, (4) Lessons learned for ERA from a commercialized GM plant, (5) Monitoring of unintended release of Bt maize in Mexico. The new thematic structure facilitates a more in-depth discussion of the presentations related to a specific topic, and the conclusions to be drawn are thus more consistent. Each moderator agreed to take responsibility for summarizing one or more themes and to prepare the respective report.