Woodlouse Armadillidium Vulgare Research Articles

Taxon-specific ability of saprophagous soil macrofauna to reintegrate carbon from agricultural waste into soil

Saprophagous soil macroinvertebrates may potentially degrade agricultural wastes. However, it is not known, to what extent and representatives of which taxa may help reintegrating carbon from crop residues back into soil without triggering massive carbon release into the atmosphere. To tackle this problem, we conducted a three-month-long microcosm experiment with 21 different species of macrofauna (each treatment replicated four times) belonging to 13 families to test their ability to degrade wheat straw. Simultaneously CO2 release from the microcosms was measured. Five species did not survive under experimental conditions. Among the remaining 16 species, three significantly increased wheat straw decomposition with Oryctes nasicornis larvae having inflicted the highest straw mass loss (64%) in comparison with the control, where no animals were added (29%). None of the tested species increased cumulative CO2 evolution from the microcosms, while two species significantly reduced it. The reduction of carbon loss with aerobic respiration was recorded for Cetonia aurata larvae and the earthworm Dendrobaena veneta (respectively 2.5 and 2-fold in relation to the control – 53.8±4.6 mg CO2-C g−1 soil dry weight during the entire experiment). The original integrative Carbon Sequestration Index by Macrofauna (CSIM) calculated for both of the measured parameters suggests that the woodlouse Armadillidium vulgare and to a smaller extent the earthworm D. veneta appear to be the most promising organisms for industrial climate-friendly organic waste recycling in terms of survival, straw processing and simultaneous reduction of CO2 emissions from soil. Our results proved that the engagement of saprophagous macrofauna in crop residue decomposition is a viable technique of carbon reincorporation into the soil. It is accompanied with CO2 release mitigation into the atmosphere.

Reproductive senescence and parental effects in an indeterminate grower.

Reproductive senescence is the decrease of reproductive performance with increasing age and can potentially include trans-generational effects as the offspring produced by old parents might have a lower fitness than those produced by young parents. This negative effect may be caused either by the age of the father, mother or the interaction between the ages of both parents. Using the common woodlouse Armadillidium vulgare, an indeterminate grower, as a biological model, we tested for the existence of a deleterious effect of parental age on fitness components. Contrary to previous findings reported from vertebrate studies, old parents produced both a higher number and larger offspring than young parents. However, their offspring had lower fitness components (by surviving less, producing a smaller number of clutches or not reproducing at all) than offspring born to young parents. Our findings strongly support the existence of trans-generational senescence in woodlice and contradict the belief that old individuals in indeterminate growers contribute the most to recruitment and correspond thereby to the key life stage for population dynamics. Our work also provides rare evidence that the trans-generational effect of senescence can be stronger than direct reproductive senescence in indeterminate growers.

The crustacean Armadillidium vulgare (Latreille, 1804) (Isopoda: Oniscoidea), a new promising model for the study of cellular senescence

AbstractSenescence, the decline of physiological parameters with increasing age, is a quasi-ubiquitous phenomenon in the living world. The observed patterns of senescence, however, can markedly differ across species and populations, between sexes, and even among individuals. To identify the drivers of this variation in senescence, experimental approaches are essential and involve the development of tools and new study models. Current knowledge of the senescence process is mostly based on studies on vertebrates and the main information about senescence in invertebrates is mostly limited to model organisms such as Caenorhabditis elegans or Drosophila melanogaster. In this context, we tested whether biomarkers of vertebrate ageing could be used to study senescence in a new invertebrate model: the common woodlouse Armadillidium vulgare (Latreille, 1804). More specifically, we looked for the effect of age in woodlouse on three well-established physiological biomarkers of ageing in vertebrates: immune cells (cell size, density, and viability), β-galactosidase activity, and the gene expression of telomerase reverse transcriptase (TERT), an essential subunit of telomerase protein. We found that the size of immune cells was higher in older individuals, whereas their density and viability decreased, and that the β-galactosidase activity increased with age, whereas the TERT gene expression decreased. These findings demonstrate that woodlouse displays age-related changes in biomarkers of vertebrate senescence, with different patterns depending on gender. The tools used in studies of vertebrate senescence can thus be successfully used in studies of senescence of invertebrates such as the woodlouse. The application of commonly used tools to new biological models offers a promising approach to assess the diversity of senescence patterns across the tree of life.

Survival capacity of the common woodlouse Armadillidium vulgare is improved with a second infection of Salmonella enterica

Immune priming has been widely observed in invertebrates. However, this phenomenon remains incompletely characterized concerning the time course of the protective effect and immune responses. Here, we investigated the existence of a protective effect in the terrestrial isopod Armadillidium vulgare against a second infection with the bacterium Salmonella enterica. We primed animals with a low dose of living bacteria or with the bacterial culture medium. One day, 7 or 15 days later, we measured haemocytes concentration and viability and we injected a LD50 of living S. enterica to monitor the survival rates of infected individuals. We show that A. vulgare is better protected (i.e. survival improvement) upon secondary infection when it has encountered S. enterica 7 days before. This protection, which tends to persist 15 days after the first infection, is however not observed when the priming is performed only 1 day before the LD50 injection. Besides providing a new invertebrate example, we highlight this protection is dynamic and may be partly due to a high haemocyte production.

Pillbug, Roly-Poly, Woodlouse Armadillidium vulgare (Latreille) (Malacostraca: Isopoda: Armadillidiidae)

The roly poly is an isopod, a type of non-insect arthropod also known as a terrestrial crustacean. It is called a roly-poly because it rolls into ball when disturbed. This defensive behavior also makes it look like a pill, which is why it is sometimes called a pillbug. In Europe, the name woodlouse is used for both roly polies and sowbugs because these arthropods are frequently found under logs. Roly polies are nocturnal, though they may be found during the day in the soil or under debris. They are beneficial in the garden or landscape. This 3-page fact sheet about the humble roly poly was written by Julie A. Franklin, Morgan A. Byron, and Jennifer Gillett-Kaufman and published by the UF Department of Entomology and Nematology, August 2015. (Photo by James Castner, University of Florida)
 EENY630/IN1099: Pillbug, Roly-Poly, Woodlouse Armadillidium vulgare (Latreille) (Malacostraca: Isopoda: Armadillidiidae) (ufl.edu)

Strength of the pathogenicity caused by feminizing Wolbachia after transfer in a new host: Strain or dose effect?

The alphaproteobacteria Wolbachia pipientis are among the most common and widespread symbionts in the animal world. Their vertical transmission mode is predicted to favour genotypes with low virulence. On the contrary, horizontal transfers of Wolbachia from one host to another have been shown to possibly increase the symbiont virulence. This situation has been previously described when two feminizing Wolbachia strains, wVulC and wVulM, from the ovaries of the woodlouse Armadillidium vulgare were introduced into another woodlouse named Porcellio dilatatus. These two Wolbachia strains induced severe symptoms and eventually caused the death of the recipient host. However, symptoms and death appeared sooner with wVulC than with wVulM. To know whether this difference was due to variation in the dose of infection or a difference in virulence between the two Wolbachia strains, we performed controlled and gradual doses of injection with wVulC and wVulM in P. dilatatus. We showed that the two strains differed intrinsically in their virulence against P. dilatatus and that their virulence is related to the injection dose. Moreover, we showed that wVulC reached higher concentrations in the recipient host than wVulM suggesting a potential link between the bacterial titers and the levels of virulence. We also addressed the impact of the tissue source of the Wolbachia used for the transinfection and demonstrated that Wolbachia transinfected via hemolymph colonized the body of the recipient more quickly and caused accelerated symptoms compared to Wolbachia introduced via a crushed ovaries suspension.

Horizontal transfers of feminizing versus non‐feminizing Wolbachia strains: from harmless passengers to pathogens

The endosymbiont Wolbachia pipientis infects various hosts in which it navigates vertically from mothers to offspring. However, horizontal transfers of Wolbachia can occur between hosts. The virulence of the horizontally acquired Wolbachia can change in the new host as it has been illustrated by the case of the feminizing strain wVulC from the woodlouse Armadillidium vulgare that turns to a pathogen when introduced into Porcellio dilatatus dilatatus. In the present study, we aim to show whether symbiotic traits, such as (i) host sex manipulation and (ii) colonization patterns, which differ between eight isopod Wolbachia strains, are connected to their virulence towards the recipient host P. d. dilatatus. Among the transferred Wolbachia, some feminizing strains gradually differing in feminizing intensity in their native hosts induced different levels of pathogenicity to P. d. dilatatus. Not a single feminizing strain passed vertically with high titres to the next generation. The non-feminizing Wolbachia strains, even if they reached high densities in the host, did not impact host life-history traits and some vertically passed with high titres to the offspring. These results suggest that a potential link between the manners Wolbachia manipulates its native host reproduction, its virulence and its ability to vertically infect the offspring.

The effect of external marking on the behaviour of the common pill woodlouse Armadillidium vulgare

Zoologists distinguish individual animals using marking techniques. Generally they test the potential influence of marking on survival only; the influence on behaviour is usually neglected. We evaluated the influence of two external marking techniques (nail polish and queen-bee marker) on the behaviour of common pill woodlouse, Armadillidium vulgare. The behaviour was examined from two points of view: (1) activity during 24 hours and (2) specific expressions of behaviour (exploring, feeding, resting and hiding) over a 24 hour period. We compared behaviour among woodlice marked with nail polish and queen-bee marker with the unmarked control group during a nine-day experiment. Although we did not find any influence of marking on survival, there was an evident influence on behaviour in most cases. Generally, in the groups of marked individuals of Armadillidium vulgare there were large differences observed against the control group in the overall activity. Activity of marked individuals was significantly reduced and they preferred hiding. The influence of polish and marker on the overall frequencies of behavioural categories was evident, mainly in feeding, resting and hiding. The influence on the frequency of exploring was significant in the polish marked group only.

Effect of introduced exotic tree litter on consumption patterns of the introduced exotic isopod Armadillidium vulgare

The feeding preferences and consumption rates of the exotic woodlouse Armadillidium vulgare were measured on litters from the autochthonous Rio Grande cottonwood and the exotic Russian olive. When offered a choice between five litter types (green and shed leaves of Russian olive and cottonwood, and fruits of Russian olive), the isopods consumed more of the exotic plant litter (green leaves and the fruits of Russian olive) than of the autochthonous cottonwood. When offered only one diet, consumption rates of green leaves and fruits of Russian olive were again the highest. Feeding was related with N and C:N ratios of the litter, with highest consumption of diets with low and high C:N ratios and lowest consumption of diets with intermediate C:N ratios. The N content was highest (and C:N ratios lowest) on isopods feeding shed leaves of Russian olive. Growth was not related to the amount or the C:N ratio of the food consumed, being highest for animals feeding on shed leaves of cottonwood. Our results show that the consumption patterns and the C and N body composition of the exotic woodlouse A. vulgare are altered when fed with Russian olive litter, suggesting that the replacement of the autochthonous cottonwood by the exotic Russian olive may negatively affect the maintenance of the population of these exotic detritivores in the area.

Male Mating Success During Parturial Intermoults in the Terrestrial Isopod Armadillidium Vulgare Revealed by the Use of a Microsatellite Locus

Abstract We investigated the value of microsatellite DNA markers to improve our knowledge of mating strategy with inference to sperm competition in particular, in the woodlouse Armadillidium vulgare. In terrestrial isopods, mature females develop a brood pouch or marsupium before egg laying, the pouch being formed by overlapping oostegites during a special moult called parturial moult. Under laboratory conditions, we show that Armadillidium vulgare females are able to mate during parturial intermoults, even in the presence of a physical barrier such as that represented by the ventral marsupial plates. Our results reveal that the contribution of a second male mating with a female between two parturial moults could represent up to 50% of the paternal alleles in the brood (mean = 28% ± 15). This contrasts with data reported in the literature concerning closely related woodlice species, where males are suggested not to be able to mate with the female during the parturial intermoult period.

Armadillidin: a novel glycine-rich antibacterial peptide directed against gram-positive bacteria in the woodlouse Armadillidium vulgare (Terrestrial Isopod, Crustacean)

We report the isolation and the characterization of a novel antibacterial peptide from hemocytes of the woodlouse Armadillidium vulgare, naturally infected or uninfected by Wolbachia, an intracellular Gram-negative bacterium. This molecule displays antibacterial activity against Gram-positive bacteria despite its composition which classes it into the glycine-rich antibacterial peptide family, usually directed against fungi and Gram-negative bacteria. The complete sequence was determined by a combination of Edman degradation, mass spectrometry and cDNA cloning using a hemocyte library. The mature peptide (53 residues) has a 5259 Da molecular mass and is post-translationally modified by a C-terminal amidation. This peptide is characterized by a high level of glycine (47%) and a fivefold repeated motif GGGFH(R/S). As no evident sequence homology to other hitherto described antibacterial peptides has been found out, this antibacterial peptide was named armadillidin. Armadillidin is constitutively expressed in hemocytes and appears to be specific of A. vulgare.

The glycosylated androgenic hormone of the terrestrial isopod Porcellio scaber (Crustacea)

It appears from grafting experiments that the androgenic hormone (AH) of terrestrial isopods has a narrow species-specificity [J. Crust. Biol. 19 (1999) 684], even if AH of different species shared common epitopes [Gen. Comp. Endocrinol. 125 (2002) 218]. To date only the glycosylated AH of the woodlouse Armadillidium vulgare has been deciphered by us [Ann. New York Acad. Sci. 839 (1998) 111; Eur. J. Biochem. 262 (1999) 727] and [Biochem. Biophys. Res. Commun. 264 (1999) 419] have confirmed the primary structure of this protein. We reported in the present paper the characterization of the AH in another species Porcellio scaber by a combination of microsequencing, mass spectrometry, and molecular cloning. We found only one glycoform which consisted of two peptide chains, A and B, of 31 and 44 amino acids, respectively, with A chain carrying on Asn 18 a N-glycosylated moiety, size of which has been determined by MALDI-MS measurements. The expected structure of the glycosylation was proposed. The deduced amino acid sequence of the AH precursor was mainly identical to the one obtained independently by another group [Zool. Sci. 20 (2003) 75]. We also showed that AH gene is exclusively expressed in androgenic glands. Sequence comparison with A. vulgare and P. scaber (population of Japan) AH was discussed.

Localization of crustacean hyperglycemic and vitellogenesis-inhibiting hormones in separate cell types in the protocerebrum of the woodlouse Armadillidium vulgare (Crustacea, Isopoda)

We gave an accurate immunolocalization of CHH (crustacean hyperglycemic hormone) and VIH (vitellogenesis-inhibiting hormone) in the brain and the sinus gland of the woodlouse Armadillidium vulgare. The two immune sera have been respectively raised against HPLC-purified CHH and against a small peptide derived from the N-terminus of VIH. By immunocytochemistry, we showed that CHH and VIH were synthesized in different perikarya and stored in different axon endings of the sinus gland. As in other crustacean species studied to date, CHH was located in the axon endings filled with the biggest granules. Immunoblotting confirmed that VIH was stored in the sinus glands of both the female and the male. These clear localizations of CHH- and VIH-antigens do not preclude that only one peptide is released from a given type of SG endings and do not rule out that each peptide can be involved in the control of different physiological processes.

Feeding rates of the woodlouse Armadillidium vulgare on herb litters produced at two levels of atmospheric CO2.

The consumption and assimilation rates of the woodlouse Armadillidium vulgare were measured on leaf litters from five herb species grown and naturally senesced at 350 and 700µll-1CO2. Each type of litter was tested separately after 12, 30 and 45days of decomposition at 18°C. The effects of elevated CO2 differed depending on the plant species. In Medicago minima (Fabaceae), the CO2 treatment had no significant effect on consumption and assimilation. In Tyrimnus leucographus (Asteraceae), the CO2 treatment had no significant effect on consumption, but the elevated CO2 litter was assimilated at a lower rate than the ambient CO2 litter after 30days of decomposition. In the three other species, Galactites tomentosa (Asteraceae), Trifolium angustifolium (Fabaceae) and Lolium rigidum (Poaceae), the elevated CO2 litter was consumed and/or assimilated at a higher rate than the ambient CO2 litter. Examination of the nitrogen contents in these three species of litter did not support the hypothesis of compensatory feeding, i.e. an increase in woodlouse consumption to compensate for low nitrogen content of the food. Rather, the results suggest that in herbs that were unpalatable at the start of the experiment (Galactites, Trifolium and Lolium), more of the the litter produced at 700µll-1 CO2 was consumed than of that produced at 350µll-1 because inhibitory factors were eliminated faster during decomposition.

Mitochondrial DNA polymorphism, sex ratio distorters and population genetics in the isopod Armadillidium vulgare.

Two maternally inherited sex ratio distorters (SRD) impose female-biased sex ratios on the wood louse Armadillidium vulgare by feminizing putative males. These SRD are (i) an intracytoplasmic bacterium of the genus Wolbachia, and (ii) another non-Mendelian element of unknown nature: the f element. Mitochondrial DNA variation was investigated in A. vulgare field populations to trace the evolution of host-SRD relationships and to investigate the effect of SRD on host cytoplasmic polymorphism. The Wolbachia endosymbionts showed no polymorphism in their ITS2 sequence and were associated with two closely related mitochondrial types. This situation probably reflects a single infection event followed by a slight differentiation of mitochondria. There was no association between the f element and a given mitochondrial type, which may confirm the fact that this element can be partially paternally transmitted. The spreading of a maternally inherited SRD in a population should reduce the mitochondrial diversity by a hitchhiking process. In A. vulgare, however, a within-population mtDNA polymorphism was often found, because of the deficient spread of Wolbachia and the partial paternal inheritance of the f element. The analysis of molecular variance indicated that A. vulgare populations are genetically structured, but without isolation by distance.

The structure of a glycosylated protein hormone responsible for sex determination in the isopod, Armadillidium vulgare.

Two glycoforms (AH1 and AH2) of androgenic hormone, and its corresponding hormone precursor derived from HPLC-purified androgenic gland extract from the woodlouse Armadillidium vulgare were fully characterized by microsequencing and mass spectrometry. The amino-acid sequences of the two glycoforms were identical; they consist of two peptide chains, A and B, of 29 and 44 amino acids, respectively, with chain A carrying one N-glycosylated moiety on Asn18. The two chains are linked by two disulfide bridges. Glycoforms were only differentiated by the size and heterogeneity of the glycan chain. The androgenic hormone precursor (16.5 kDa) was shown to contain the sequence of chains A and B from the androgenic hormone, connected by a C-peptide (50 amino acids). These results were confirmed by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) analysis performed on a single hypertrophied androgenic gland. When injected into young females, both glycoforms of the androgenic hormone were able to override genetic sex-determination. In invertebrates, there is no other example where sex-differentiation is controlled by a protein hormone that is not synthesized by the gonads but by a special gland. A functional comparison with two other hormones which are believed to play a role in sex determination, i.e. ecdysone in insects and anti-Müllerian hormone in mammals, is presented. Work is in progress to clone and characterize the gene encoding androgenic hormone, moreover special attention is devoted to its regulatory regions, putative targets for the Wolbachia action.

Plasmid transfer between introduced and indigenous bacteria in leaf litter, soil and vermicompost as affected by soil invertebrates

Plasmid transfer between introduced strains of Pseudomonas stutzeri JM302 (pLV1013), Pseudomonas putida PAW340 (pLV1017), Pseudomonas aeruginosa PAO5 (RP4), and Enterobacter cloacae MF10 (RP4), all of them harbouring genetically modified or naturally occurring plasmids and bacteria indigenous to oak leaf litter, soddy-podzolic soil or vermicompost was monitored using non-sterile laboratory microcosms inhabited by the millipede Pachyiulus flavipes, the woodlouse Armadillidium vulgare, or the earthworms Aporrectodea caliginosa and Eisenia fetida, respectively. Plasmid transfer from genetically modified Streptococcus faecalis JH2-2 (pAMβ1) to Bacillus thuringiensis var. israelensis 1-5 under similar conditions was also followed. A recombinant conjugative plasmid (pLV1017) encoding resistance to antibiotics and expressing catechol 2,3-dioxygenase was transferred into two facultatively anaerobic gram-negative bacteria, isolated from the excrement of A. vulgare. Presumed transconjugants of the facultatively anerobic gram-negative bacteria given above were isolated from the leaves and soil (four strains) and the excrement of the invertebrates (11 strains). They carried the same plasmids and exhibited the same enzymatic activity as the respective inoculant strains; for most of them, however, DNA fingerprints were slightly different. Transfer from S. faecalis JH2-2 to B. thuringiensis var. israelensis of a naturally occurring plasmid, pAMb1, encoding resistance to erythromycin was observed in vermicompost.

Sterile intersexuality in an isopod induced by the interaction between a bacterium (Wolbachia) and the environment

An endocellular bacterium of the genus Wolbachia changes chromosomic (ZZ) males into functional females in a number of populations of the woodlouse Armadillidium vulgare. The interaction between the feminizing effect of Wolbachia and the lethal effect of high temperature on these bacteria is shown to be responsible for the appearance of high proportions of sterile intersexes (Si). Wolbachia-infected females produced an average of 15% Si when reared for 200 days under a daily thermoperiodic regime that included 4 h at 30°C. A temperature of 30°C is known to destroy Wolbachia. The Si phenotype may therefore be due to the partial destruction (or inhibition) of the feminizing bacteria by high temperature during development. This induction of a proportion of Si differs in two ways from the intersexuality induced by the conflict between the feminizing agent and a host autosomal gene. First, the genomic conflict does not lead to the production of numerous Si, and second, the temperature-dependent production of Si is a sporadic event induced by the environment, rather than being a heritable trait. The overproduction of sterile offspring at high temperatures can result inWolbachia-infected females of A. vulgare suffering a loss of fitness.

Photoperiodism and seasonal breeding in aquatic and terrestrial Eumalacostraca

Summary Seasonal reproduction in Crustacea is synchronized by environmental cues (e.g., temperature and photoperiod). Two types of responses occur: (1) in reptant decapods, short photoperiods and low temperatures are necessary to induce vitellogenesis; (2) in natant decapods and in most peracarids, both high temperatures and long photoperiods promote the onset of reproduction. In this context studies on the biological clocks involved in photoperiodic time measurement were performed on two crustaceans: the ditch shrimp Palaemonetes varians and the woodlouse Armadillidium vulgare, which is distributed worldwide outside polar and intertropical areas. The most important feature of the photoperiodic response curves was the same mean lag time in response to photophase length ≤ 12 h: short photophases delayed breeding but did not prevent it (i.e., “short-day effect”). On the contrary, long photophases promoted the onset of reproduction (i.e., “long-day effect”) differently in P. varians and A. vulgare. In A. vul...

The spread of parasitic sex factors in populations of Armadillidium vulgare Latr (Crustacea, Oniscidea): effects on sex ratio

Summary - In the woodlouse Armadillidium vulgare, the basis of sex determination is under the control of heterochromosomes (ZZ males and WZ females). However, in most populations, sex is determined by 2 maternally-transmitted parasitic sex factors (PSF). One is a Wolbachia-like bacterium (F), the other (f) is probably a sequence of F DNA unstably integrated in the A vulgare genome. Both PSF are feminizing factors, which transform genetic males into functional neo-females (ZZ+F or ZZ+f). The propagation of these PSF was investigated by introducing neo-females into artificial populations initially formed of genetic females. In the majority of these populations, F bacteria were unable to spread, while f seemed to have a higher invasive power. These results contrast with theoretical data: owing to the high transmission rate of PSF, their frequency should increase in populations where they have appeared. Based on the observation that PSF in populations have a behaviour similar to that of genes with selective advantages, a new approach was proposed to describe the spreading of PSF in limited-sized populations. This model showed that even a high transmission rate and in the absence of nuclear genes conferring resistance in A vulgare, PSF can be lost from populations. The equilibrium probability of their disappearance depends on both the number of neo-females infesting the population and the transmission rate of PSF. These theoretical data, combined with the presence of genes repressing the feminizing effects of PSF, may explain part of the variability observed in sex determining systems and sex ratios of natural populations.