Response of the terrestrial isopod Porcellio scaber to lipopolysaccharide challenge after microplastic and insecticide exposure.

The exposure of organisms to microplastics could compromise their ability to cope with other environmental stressors, such as infections. In this context, we investigated the effects of a 14-day exposure of the terrestrial isopod Porcellio scaber to tire particles in soil (1.5 % w w-1 dry weight) on the organisms' response to a secondary exposure, i.e., injection of the bacterial endotoxin lipopolysaccharide. In addition, the insecticide chlorpyrifos (2 mg kg-1 dry weight) was tested as a positive control. The survival and immune response of P. scaber was assessed at the end of the 7- and 14-day primary exposure and two days after the secondary exposure, by analyzing selected haemolymph immune parameters (total haemocyte count, differential haemocyte count, and haemocyte viability). No change in survival was observed after primary exposure of P. scaber to tire particles or chlorpyrifos. However, primary exposure to chlorpyrifos triggered a strong activation of the immune response, which was not the case following exposure to the tire particles. Further injection of lipopolysaccharide into the body did not affect the survival of animals exposed to tire particles or chlorpyrifos, while a strong immunomodulatory change was observed, particularly with chlorpyrifos, and to some extent, tire particles. Based on these results, we conclude that exposure of P. scaber to tire particles or chlorpyrifos has no significant effect on the susceptibility of the organism to lipopolysaccharide in terms of their mortality, but primary exposure to an insecticide significantly modulates the immune response of the organisms to a second stressor. We discuss the "stress on stress" approach for testing low-toxic substances, such as microplastics, where an environmentally realistic exposure is followed by a secondary exposure.

Proteomics of the haemolymph of the terrestrial crustacean Porcellio scaber reveals components of its innate immunity under baseline conditions.

The terrestrial crustacean Porcellio scaber is an established test organism in environmental research. We analysed the haemolymph proteome of P. scaber using a classical proteomic approach based on one-dimensional gel electrophoresis and tandem mass spectrometry. Using a publicly available protein database and our P. scaber transcriptome data, we have identified 76 proteins involved in cytoskeleton formation, protein degradation, vesicular transport, genetic information processing, detoxification, carbohydrate and lipid metabolism reflecting haemocyte metabolic activity, active intracellular transport, and intercellular communication. Compared with the data reported for other crustaceans, 28 of these P. scaber proteins have been linked to its immunity, among them hemocyanin, α-2-macroglobulin, phenoloxidase 3, superoxide dismutase, glutathione S-transferase, haemolymph clottable protein, and histones H4 and H2B. Our results thus provide a firm base for studying the innate immune response of P. scaber at the level of the haemolymph proteome. This knowledge is of particular importance in ecotoxicity studies with various environmental stressors where understanding physiological changes is important to reveal possible modes of action.

Tire microplastics exposure in soil induces changes in expression profile of immune-related genes in terrestrial crustacean Porcellio scaber.

Tire particles pose a potential threat to terrestrial organisms because they are deposited in large quantities in the soil by tire wear abrasion, and moreover their chemical complexity poses an additional risk. Microplastics can affect several physiological processes in organisms, including those related to immunity. Therefore, we investigated the expression profile of selected immune-related genes (MnSod, Manganese Superoxide dismutase; Cat, Catalase; CypG, Cyclophilin G; Nos, Nitric oxide synthase; Ppae2a, Prophenoloxidase-activating enzyme 2a; Dscam, Down syndrome cell adhesion molecule; Myd88, Myeloid-differentiation factor 88; Toll4, Toll-like receptor 4; Mas-like, Masquerade-like protein) in haemocytes and the digestive gland hepatopancreas of terrestrial crustacean Porcellio scaber after two different time exposures (4 and 14 days) to tire particles in soil. Our results reveal for the first time the response of P. scaber after microplastic exposure at the transcriptome level. We observed time- and tissue-dependent changes in the expression of the analysed genes, with more pronounced alterations in haemocytes after 14 days of exposure. Some minor changes were also observed in hepatopancreas after 4 days. Changes in the expression profile of the analysed genes are a direct indication of a modulated immune status of the test organism, which, however, does not represent an adverse effect on the test organism under the given conditions. Nevertheless, the question remains whether the observed change in immune status affects the immunocompetence of the test organism.

Time-dependent immune response in Porcellio scaber following exposure to microplastics and natural particles.

Microplastics are very common contaminants in the environment. Despite increasing efforts to assess the effects of microplastics on soil organisms, there remains a lack of knowledge on how organisms respond to diverse types of microplastics after different exposure durations. In the present study, we investigated the immune response of the terrestrial crustacean Porcellio scaber exposed to the two most common microplastic particles in the environment: polyester fibres and tyre particles. We also tested two natural particles: wood dust and silica powder, with all treatments performed at 1.5% w/w. The response of P. scaber was evaluated at the level of the immune system, and also the biochemical, organism and population level, after different exposure durations (1, 2, 4, 7, 14, 21 days). These data reveal dynamic changes in the levels of some immune parameters shortly after exposure, with a gradual return to control values. The total number of haemocytes was significantly decreased after 4 days of exposure to tyre particles, while the proportion of different haemocyte types in the haemolymph was altered shortly after exposure to both polyester fibres and tyre particles. Moreover, 7 days of exposure to tyre particles resulted in increased superoxide dismutase activity in the haemolymph, while metabolic activity in whole woodlice (measured as electron transport system activity) was increased after exposure for 7, 14 and 21 days. In contrast, the natural particles did not elicit any significant changes in the measured parameters. Survival and feeding of P. scaber were not altered by exposure to the microplastics and natural particles in soil. Overall, this study defines a time-dependent transient immune response of P. scaber, which indicates that immune parameters represent sensitive biomarkers of exposure to microplastics. We discuss the importance of using natural particles in studies of microplastics exposure and their effects.

Avoidance behaviour toxicity tests should account for animal gregariousness: a case study on the terrestrial isopod Porcellioscaber.

Avoidance behaviour enables woodlice to escape suboptimal environmental conditions and to mitigate harmful effects of pollutants. However, several studies have shown that at least in some woodlice species the tendency to aggregate can lead to suboptimal responses as the between-conspecific attraction can outweigh the aversive stimuli. The present study evaluated the influence of gregariousness on the behaviour of Porcellioscaber in a heterogeneously polluted environment. The hypothesis was that the tendency for aggregation outweighs the tendency for exploratory activity, therefore animals in groups will be less active. Consequently, this will affect their avoidance of polluted environmental patches. To test this hypothesis, isolated individuals or pairs of individuals were monitored in free-choice arenas where animals could select between uncontaminated and pyrethrin-contaminated soils. Animals were video recorded for 3 h in darkness using infrared light and analysed for avoidance behaviour and locomotor activity. In general, isolated animals were more locomotory active and avoided the contaminated soil more than paired animals. It can be concluded that aggregation behaviour suppresses exploratory behaviour and consequently also the avoidance of polluted environments. This should be accounted for when interpreting results of avoidance tests with groups of gregarious animals, which may underestimate the effect of pollutants.

Daily and seasonal time partitioning in surface activity of Porcellioalbinus (Isopoda, Oniscidea) in the arid region of Zarat (Gabes, Tunisia).

The terrestrial isopod Porcellioalbinus is a burrowing species, dwelling in the desert of south Tunisia. Field studies were carried out in the coastal area of Zarat, Tunisia, to examine the surface activity rhythm of P.albinus in relation to daily and seasonal variations in environmental conditions. The activity of P.albinus was followed once a month, from November 2012 through October 2013. Hourly capture frequency was compared across the different seasons of the year. Porcellioalbinus is a strictly nocturnal species showing a nycthemeral rhythm regulated by the rhythmic and natural variations of the duration of the dark period. A positive correlation is observed between the circadian rhythm of the locomotor activity of the species and the duration of the dark period outside its burrow, P.albinus has a single daily activity peak. Individuals concentrated their activity in the first part of the night in winter and in the second part in summer. This peak is more spread out in spring and autumn. The differences in the activity rhythm of P.albinus between different seasons may be determined by two important factors, namely temperature and the length of the dark period.

Inventory of the terrestrial isopods in Belgium (2011-2020).

This data paper describes a recent and spatially complete inventory of the terrestrial isopods of Belgium between 2011 and 2020. During these 10 years every 10 × 10 km² cell of the Universal Transverse Mercator (UTM) grid in Belgium (373 grid cells) was visited in search for terrestrial isopods. Inventories covered different habitat types in every grid cell such as forest, wetlands or stream sides, and urban areas. Most of the dataset records were obtained by hand-collection methods such as turning stones and dead wood, or by sieving litter and through casual observations. These inventories were carried out by specialists from Spinicornis, the Belgian Terrestrial Isopod Group. Their data is complemented with pitfall trap data from scientific projects and verified citizen science data collected via waarnemingen.be and observations.be from the same time period. This resulted in 19,406 dataset records of 35 terrestrial isopod species. All dataset records are georeferenced using the centroid of their respective 5 × 5 km² UTM grid cell. The dataset is published as open data and available through the Global Biodiversity Information Facility (GBIF). Direct link to the dataset: https://doi.org/10.15468/mw9c66.

Bug cemetery: a case study of terrestrial isopod distribution on a brick wall in the Czech Republic.

Although terrestrial isopods (Oniscidea) are primarily soil- and surface-dwelling invertebrates, they can also be found on tree trunks and walls. This study evaluated distribution patterns of terrestrial isopods on a brick wall during the first hours of night in autumn. Four species of terrestrial isopods were recorded with Armadillidiumversicolor being the dominant one. Terrestrial isopods were distributed from ground level up to a height of 2 m, but preferred a 70-80 cm height band. The highest number of active individuals was observed 3 h after astronomical dusk. Potential predators of terrestrial isopods were abundant during the same time and at the same height.

New records of Trachelipusvespertilio (Budde-Lund, 1896) (Isopoda, Oniscidea) with a description of the male.

Background: The existing descriptions of the woodlouse Trachelipusvespertilio are based on a single female collected in Croatia in the nineteenth century. No further information on the occurrence of this species has been reported in published literature and the morphology of the male, which may offer additional reliable diagnostic characters, has remained unknown. New information: On the basis of new material collected in Slovenia, a description of the male morphology of T.vespertilio is provided along with a species diagnosis. The rediscovery of this woodlouse after more than a century extends its distribution range to Slovenia.

Urban effects on saprophagous macroarthropods are mainly driven by climate: A global meta-analysis.

Macrodecomposers provide important ecosystem services even in human dominated habitats including urban ecosystems, but the effect of urban land conversion on their species diversity and abundance has not been explored at global scale. Here, we present the first meta-analysis to quantify the general response of two major arthropod taxa, terrestrial isopods and millipedes to urbanization and to reveal the underlying mechanisms. Climatic (temperature, precipitation, growing season length), edaphic (pH, organic carbon, CaCO3 and clay content of surface soils), urban (population density, city age, vegetation cover and mean actual evapotranspiration) parameters and methods of study (duration, sampling technique, replications) were used as moderators. We used a hierarchical meta-analytic approach to consider the dependence of multiple effect sizes obtained from one study. Altogether 156 paired observations were extracted from 59 urban studies conducted between 1980 and 2020. Urbanization had a negative effect on species diversity (species richness and Shannon index) of both macroarthropod taxa. However, both the direction and strength of their abundance response varied to a greater extent, resulting in a neutral effect of urban disturbance on them. The key drivers influencing the urban effects on macroarthropods were mean annual temperature and precipitation, absolute minimum temperature and length of growing season. The study also highlighted the importance of sampling methods: direct sampling (hand collecting) resulted in stronger urban effects presumably due to several sources of sampling bias. Our global synthesis highlighted that urbanization is a threat to soil arthropods, particularly to litter-dwelling detritivores, which potentially alters plant residue processing and ultimately soil biogeochemical cycles.

Long Term Exposure to Virgin and Recycled LDPE Microplastics Induced Minor Effects in the Freshwater and Terrestrial Crustaceans Daphnia magna and Porcellio scaber.

The effects of microplastics (MP) are extensively studied, yet hazard data from long-term exposure studies are scarce. Moreover, for sustainable circular use in the future, knowledge on the biological impact of recycled plastics is essential. The aim of this study was to provide long-term toxicity data of virgin vs recycled (mechanical recycling) low density polyethylene (LDPE) for two commonly used ecotoxicity models, the freshwater crustacean Daphnia magna and the terrestrial crustacean Porcellio scaber. LDPE MP was tested as fragments of 39.8 ± 8.82 µm (virgin) and 205 ± 144 µm (recycled) at chronic exposure levels of 1-100 mg LDPE/L (D. magna) and 0.2-15 g LDPE/kg soil (P. scaber). Mortality, reproduction, body length, total lipid content, feeding and immune response were evaluated. With the exception of very low inconsistent offspring mortality at 10 mg/L and 100 mg/L of recycled LDPE, no MP exposure-related adverse effects were recorded for D. magna. For P. scaber, increased feeding on non-contaminated leaves was observed for virgin LDPE at 5 g/kg and 15 g/kg. In addition, both LDPE induced a slight immune response at 5 g/kg and 15 g/kg with more parameters altered for virgin LDPE. Our results indicated different sublethal responses upon exposure to recycled compared to virgin LDPE MP.

Stressor-Dependant Changes in Immune Parameters in the Terrestrial Isopod Crustacean, Porcellio scaber: A Focus on Nanomaterials.

We compared the changes of selected immune parameters of Porcellio scaber to different stressors. The animals were either fed for two weeks with Au nanoparticles (NPs), CeO2 NPs, or Au ions or body-injected with Au NPs, CeO2 NPs, or lipopolysaccharide endotoxin. Contrary to expectations, the feeding experiment showed that both NPs caused a significant increase in the total haemocyte count (THC). In contrast, the ion-positive control resulted in a significantly decreased THC. Additionally, changes in phenoloxidase (PO)-like activity, haemocyte viability, and nitric oxide (NO) levels seemed to depend on the stressor. Injection experiments also showed stressor-dependant changes in measured parameters, such as CeO2 NPs and lipopolysaccharide endotoxin (LPS), caused more significant responses than Au NPs. These results show that feeding and injection of NPs caused an immune response and that the response differed significantly, depending on the exposure route. We did not expect the response to ingested NPs, due to the low exposure concentrations (100 µg/g dry weight food) and a firm gut epithelia, along with a lack of phagocytosis in the digestive system, which would theoretically prevent NPs from crossing the biological barrier. It remains a challenge for future research to reveal what the physiological and ecological significance is for the organism to sense and respond, via the immune system, to ingested foreign material.

Modulations of immune parameters caused by bacterial and viral infections in the terrestrial crustacean Porcellio scaber: Implications for potential markers in environmental research.

The terrestrial crustacean Porcellio scaber (Crustacea: Isopoda) is an established invertebrate model in environmental research. Preceding research using isopods did not widely use immune markers. In order to advance their use in research, knowledge of the reference values in control animals as well as variations during infections is of importance. This study presents, for the first time, the morphology, and ultrastructure of the three main haemocyte types of Porcellio scaber as semigranulocytes (SGCs), granulocytes (GCs), and hyalinocytes (HCs), with the latter having two subtypes, using various light and electron microscopy approaches. The modulation of selected immune cellular and humoral parameters of P. scaber in symptomatic phases of Rhabdochlamydia porcellionis and Iridovirus IIV-31 infections is presented. A clear difference in the immune responses of bacterial and viral infections was shown. Remarkable changes in total haemocyte count (THC) values and the proportions of three different haemocyte types were found in animals with a viral infection, which were not as significant in bacterially infected animals. Modified NO levels and SOD activity were more pronounced in cases of bacterial infection. Knowledge of the morphological and ultrastructural features of distinct haemocyte types, understanding the baseline values of immune parameters in control animals without evident symptoms of infection, and the influence that infections can have on these parameters can serve as a basis for the further use of P. scaber immune markers in environmental research.

Woodlice change the habitat use of spiders in a different food chain.

BACKGROUND: In old field systems, the common woodlouse may have an indirect effect on a nursery web spider. Woodlice and nursery web spiders feed in different food chains, yet previous work demonstrated that the presence of woodlice is correlated with higher predation success by nursery web spiders upon their grasshopper prey. This finding suggested a new hypothesis which links two seemingly disparate food chains: when woodlice are present, the spider predator or the grasshopper prey changes their location in the vegetative canopy in a way that increases their spatial overlap and therefore predation rate. However, warming temperatures may complicate this phenomenon. The spider cannot tolerate thermal stress, meaning warming temperatures may cause the spider to move downwards in the vegetative canopy or otherwise alter its response to woodlice. Therefore, we would expect warming and woodlice presence to have an interactive effect on predation rate. METHODS: We conducted behavioral experiments in 2015, 2017, and 2018 to track habitat domains-the use of the vegetative canopy space by grasshoppers and spiders-in experimental cages. Then, we used three models of spider movement to try to explain the response of spiders to woodlice: expected net energy gain, signal detection theory, and individual-based modelling. RESULTS: Habitat domain observations revealed that spiders shift upward in the canopy when woodlice are present, but the corresponding effect on grasshopper prey survival was variable over the different years of study. Under warming conditions, spiders remained lower in the canopy regardless of the presence of woodlice, suggesting that thermal stress is more important than the effect of woodlice. Our modelling results suggest that spiders do not need to move away from woodlice to maximize net energy gain (expected net energy gain and signal detection theory models). Instead spider behavior is consistent with the null hypothesis that they move away from unsuccessful encounters with woodlice (individual-based simulation). We conclude that mapping how predator behavior changes across biotic (e.g. woodlouse presence) and abiotic conditions (e.g. temperature) may be critical to anticipate changes in ecosystem dynamics.

Remediation of heavy metals and enhancement of fertilizing potential of a sewage sludge by the synergistic interaction of woodlice and earthworms.

Woodlice and the earthworm alone or in combination were used to improve physical properties, nutrient release, and heavy metals stabilization during composting of sewage sludge. Chemical properties of raw sludge (IS) were compared to those of composted sludge (CS), sludge + earthworms (VS), sludge + woodlice (WS), and sludge + earthworms + woodlice (VWS) after 50 and 100 days of composting. Physical properties and heavy metals accumulation by the studied fauna was determined after 100 days of composting. Highest proportions of fine particles, porosity and water holding capacity were in the VWS treatment, electrical conductivity, ash content, inorganic N and the total concentrations of P, K, Ca and Mg increased, whereas the pH value, the organic C, C:N ratio and humic acids content decreased in following order: VWS > VC > WS > CS > IS. Total concentrations of Pb, Cd and Ni in composted sludge were lower whereas concentrations of Zn, Cu and Mn were higher than in raw sludge, and woodlice showed higher bioconcentration factor (BCFs) than earthworm for all heavy metals in all treatments. We concluded that woodlice and earthworms synergistically improved the physicochemical properties of the compost and enhanced its potential use as amendment in agricultural soils.

Horizontal and vertical exploration in woodlice: A dual-process model.

Woodlice placed in an unknown experimental enclosure typically run (horizontal exploration) and rear up on the enclosure's walls (vertical exploration). Previous findings with Porcellio scaber indicate that these two behaviors have an opposite temporal distribution and show differential sensitivity to rotation-induced physiological stress. It is argued that the dual-process theory of habituation and sensitization can serve as a basis to account for horizontal and vertical exploratory activities in woodlice. A model is presented. This model could help identify individual variability in "temperament" and to isolate its effects in various behavioral tasks in woodlice and perhaps other invertebrate species.

A molecular phylogeny of Porcellionidae (Isopoda, Oniscidea) reveals inconsistencies with present taxonomy.

Porcellionidae is one of the richest families of Oniscidea globally distributed but we still lack a comprehensive and robust phylogeny of the taxa that are assigned to it. Employing five genetic markers (two mitochondrial and three nuclear) we inferred phylogenetic relationships among the majority of Porcellionidae genera. Phylogenetic analyses conducted via Maximum Likelihood and Bayesian Inference resulted in similar tree topologies. The mtDNA genes cytochrome oxidase I (COI) and 16s rRNA (16s) were used for clade dating using previously published mutation rates. Our results provide evidence against the monophyly of both Porcellionidae and the largest genus of the family Porcellio. These results are compared to previous published work based on morphological evidence. The genera Leptotrichus and Brevurus are not grouped with the rest of Porcellionidae whereas Agnaridae are grouped with part of Porcellionidae. Armadillidium and Schizidium (Armadillidiidae) occupy a basal position on the phylogenetic tree. Even though the African genera Tura and Uramba (distributed in East Africa) are grouped together there is no general geographical pattern in other sub-clades. Additional taxonomic issues that arise in this work such as the assignment of the recently described genus Levantoniscus, arealso discussed. The status of Porcellionidae should be further revised and morphological characters traditionally used in Oniscidea taxonomy should be reconsidered in view of molecular evidence. The origin of the monophyletic clade within Porcellionidae as indicated in the present work is dated back to the Oligocene (~32 mya).

Effect of agricultural practices on terrestrial isopods: a review.

Terrestrial isopods (approximately 3700 known species in the world) are encountered in temperate and tropical regions, from the seashore to high altitudes and from floodplain forests to deserts. They are known to contribute to soil biodiversity. Environmental factors and anthropogenic actions, particularly land use changes such as primarily agricultural practices, and urbanization affect soil biodiversity and their functions. Human practices, such as soil tillage, pesticide application, chemical pollution, along with soil acidification adversely affect isopod abundance and diversity. It is thus important to recognise the vital contributions of soil biodiversity in support of environmental quality protection through maintaining soil functions and their significance to sustainable land use. This review will also deal with recent studies attempting to evaluate the impact of returning to an environmentally friendly agriculture by restoring refuge habitats such as grass strips, hedges, and woodlands for terrestrial isopods.

The role of urban forest patches in maintaining isopod diversity (Oniscidea).

Compositional changes in natural communities associated with anthropogenic influence often lead to localised extinctions and biodiversity loss. Soil invertebrates are also threatened by urbanisation due to habitat fragmentation, vegetation changes and management, soil alteration, degradation, and disappearing shelter sites. The aim was to assess terrestrial isopod (Oniscidea) assemblages in differently degraded urban forest patches of a metropolitan area (Budapest, Hungary). Study sites were compared by their species richness, composition and the relevant background factors (soil properties, dead wood, litter characteristics, and canopy closure). The degree of urban disturbance was expressed using an urbanisation index (UI) based on built-up density and vegetation cover. The isopods were identified to species level, and were qualified by their habitat preference and naturalness index (TINI). Average Rarity Index (ARI), derived from TINIs provided information on the degree of naturalness/disturbance of each habitat. Altogether 14 isopod species were collected from 23 sample sites. Urbanisation indirectly affected on the composition of isopod assemblages through the quantity of dead wood and soil plasticity. ARIs and UIs of sample sites were negatively correlated. Urban patches harboured habitat generalist, synanthropic and established introduced species with low naturalness value of assemblages. Areas with no or low anthropogenic disturbance maintained stable native, autochthonous assemblages that were characteristic of rural sites in the region. Transitional zones between rural and urban habitats usually maintained a mixed isopod fauna consisting of both urban and rural elements.

Two new species of Atlantoscia Ferrara Taiti, 1981 (Isopoda: Oniscidea: Philosciidae) from southern Brazil described in the light of integrative taxonomy.

The use of molecular data in association with classical taxonomy has helped to alleviate the taxonomic impediment through the discovery, delimitation and description of new species. Terrestrial isopods are the largest suborder of Isopoda; however, there are very few active taxonomists in this group and a representative part of the terrestrial isopods world diversity remains unexplored. The genus Atlantoscia comprises five species in which diagnostic characters are few and show subtle differences among the species. The objective of this study was to delimit and describe two new species of the genus Atlantoscia from southern Brazil by using traditional taxonomy coupled with genetic information. Specimens were analyzed morphologically and by the aid of two molecular markers, mitochondrial COI and nuclear 18S rRNA. The validity of the new species Atlantoscia antennamaculata sp. nov. and Atlantoscia australis sp. nov. was corroborated, unambiguously, by morphological approach, phylogenetic analysis and species delimitation methods. Our study reinforces the fact that, despite the undeniable importance of comparative morphology in species discovery, new methods and data, particularly molecular ones, are becoming prominent and an integrative part of the taxonomy practice.