Skip to main content
SpringerLink
Log in

Detection and phylogeny of the bacteria Wolbachia in the terrestrial isopod Porcellio laevis Latr. (Crustacea, Oniscoidea), isolated from Lebna and Bizerte stations, Tunisia

  • Original Article
  • Published:
Annals of Microbiology Aims and scope Submit manuscript

Abstract

Wolbachia Hertig, 1936, is an intracytoplasmic bacterium that infects several species of arthropods by causing deterioration of host reproduction. In terrestrial isopods, Wolbachia infection can generate cytoplasmic incompatibility and feminization. This work examined two samples of Wolbachia from two different stations in Tunisia (Lebna in the North-East, and Bizerte in the North), and the study is limited to the host species Porcellio laevis Latreille, 1804. Enumeration of males and females revealed a very strong imbalance in the sex ratio in favour of females in the former population, and intersexual females in the latter. Dissection of the genital apparatus of various phenotypically normal male specimens allowed some malformations to be noted. These males were marked by the presence of an oviduct joined to the vas deferens, and one to three hypertrophied androgenic glands at the top of the testicles. Further molecular study confirmed the presence of the endosymbiont in the two populations. Sequencing of the P. laevis wsp gene from the Bizerte population revealed a high degree of identity with the Wolbachia endosymbiont of Armadillidium vulgare.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1a,b
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Achouri SM, Charfi-Cheikhrouha F (2002) Biologie et dynamique des populations de Porcellionides sexfasciatus (Crustacé, Isopode Oniscoidea). C R Biol 325:605–661

    Article  PubMed  Google Scholar 

  • Azzouna A, Grève P, Martin G (2004) Sexual differentiation in functional males with female genital apertures (♂ fga) in the woodlice Armadillidium vulgare Latr. (Isopoda, Crustacea). Gen Comp Endocrinol 138:42–49

    Article  CAS  PubMed  Google Scholar 

  • Beaumont A, Cassier P, Truchot JP, Dauça M (2006) Biologie et Physiologie Animales (cours et questions de revision). Dunod, Paris

  • Benlarbi-Ben Khedher M (2006) Etude épidémio-moléculaire et ultrastructurale de Wolbachia dans les Phlébotomes. Thèse, Université el Manar, Faculté des Sciences de Tunis

  • Ben Nasr S, Zghal F, Azzouna A (2009) Féminisation chez Porcellio laevis Latreille, 1802 (Isopode, oniscoïde) de la station de Bizerte, Tunisie (Feminization in Porcellio laevis Latreille, 1802 (Isopoda, oniscidea) from Bizerte, Tunisia. Crustaceana 82(2):219–232

    Article  Google Scholar 

  • Bordenstein SR, Wernegreen JJ (2004) Bacteriophage flux in endosymbionts (Wolbachia): infection frequency, lateral transfer, and recombination rates. Mol Biol Evol 21(10):1981–1991

    Article  CAS  PubMed  Google Scholar 

  • Bouchon D, Rigaud T, Juchault P (1998) Evidence for widespread Wolbachia infection in isopod crustaceans: molecular identification and host feminization. Proc R Soc B Biol Sci 265:1081–1090

    Article  CAS  Google Scholar 

  • Braquart-Varnier C, Grève P, Felix C, Martin G (2005) Bacteriophage WO in Wolbachia infecting terrestrial Isopods. Biochem Biophys Res Commun 337:580–585

    Article  CAS  PubMed  Google Scholar 

  • Cordaux R, Michel-Salzat A, Bouchon D (2001) Wolbachia infection in crustaceans: novel hosts and potential routes for horizontal transmission. J Evol Biol 14(2):237–243

    Article  CAS  Google Scholar 

  • Cordaux R, Michel-Salzat A, Frelon-Raimond M, Rigaud T, Bouchon D (2004) Evidence for a new feminizing Wolbachia strain in the isopod Armadillidium vulgare: evolutionary implications. Heredity 93:78–84

    Article  CAS  PubMed  Google Scholar 

  • Felix C (2004) Etude moléculaire de la bactérie intracellulaire féminisante Wolbachia chez Armadillidium vulgare (crustacé, isopode terrestre). Thèse, Université de Poitiers

  • Felsentein J (1985) Confidence limits on phylogeny: an approach using the bootstrap. Evolution 39:783–791

    Article  Google Scholar 

  • Hiroki M, Kato Y, Kamito T, Miura K (2002) Feminization of genetic males by a symbiotic bacterium in a butterfly, Eurema hecabe (Lepidoptera: Pieridae). Naturwissenschaften 89:167–170

    Article  CAS  PubMed  Google Scholar 

  • Hmaied S, Charfi-Chekhrouha F (2004) Life cycle and population dynamic of Armadillidium pelagicum Arcacangeli, 1955 (Isopoda, Oniscoidea) at Aouina. C R Biol 327:343–352

    Article  Google Scholar 

  • Johnson M, Verne S, Bouaziz K, Bouchon D (2007) The costs and advantages of a Wolbachia-Woodlice infection, a classical example of parasite trade-off. In: Proceedings of the 7th International Symposium on Biology Terrestrial Isopods; 28–31 March 2007, Tunis, Abstracts

  • Juchault P (1966) Contribution à l’étude de la différenciation sexuelle mâle chez les Crustacés Isopodes. Thèse, Université de Poitiers

  • Juchault P, Legrand JJ (1972) Croisement de néo-mâles expérimentaux chez Armadillidium vulgare Latr. (Crustacé, Isopode, Oniscoïde). Mise en évidence d’une hétérogamétie femelle. C R Acad Sci Paris 274:1387–1389

    Google Scholar 

  • Juchault P, Legrand JJ (1979) Analyse génétique et physiologique de la détermination du sexe dans une population du crustacé Oniscoïde—Armadillidium nasatum Budde-Lund. Arch Zool Exp Gén 120:25–43

    Google Scholar 

  • Juchault P, Legrand JJ (1981) Contribution à l’étude qualitative et quantitative des facteurs contrôlant le sexe dans les populations du crustacé isopode terrestre, Armadillidium vulgare Latreille, II-Populations hébergeant le facteur féminisant F (Bactéroїde intracytoplasmique). Arch Zool Exp Gén 122:65–74

    Google Scholar 

  • Juchault P, Mocquard JP (1989) Effet de l’inoculation d’une bactérie endocellulaire féminisante sur la croissance et la reproduction des femelles du crustacé Oniscoїde Armadillidium vulgare (Latr.). Conséquences possibles sur l’évolution des populations. Crustaceana 56(1):83–92

    Article  Google Scholar 

  • Juchault P, Legrand JJ, Martin G (1974) Action interspécifique du facteur épigénétique que féminisant responsable de la thélygénie et de l’intersexualité du Crustacé Armadillidium vulgare (Isopode, Oniscoïde). Ann Embryol Morphog 7(3):265–272

    Google Scholar 

  • Juchault P, Martin G, Legrand JJ (1980) Induction par la température d’une physiologie mâle chez les néo-femelles et les intersexués du Crustacé Oniscoïde Armadillidium vulgare Latr., hébergeant un bactéroïde à action féminisante. Int J Invertebr Reprod Dev 2:223–235

    Google Scholar 

  • Juchault P, Frelon M, Bouchon D, Rigaud T (1994) New evidence for feminizing bacteria in terrestrial isopods: evolutionary implications. C R Acad Sci Paris 317:325–230

    Google Scholar 

  • Legrand JJ, Juchault P (1986) Rôle des bactéries symbiotiques dans l’intersexualité, la monogénie et la spéciation chez des crustacés oniscoїdes. Boll Zool 53:161–172

    Google Scholar 

  • Lo N, Casiraghi M, Salati E, Bazzocchi C, Bandi C (2002) How many Wolbachia supergroups exist? Mol Biol Evol 19(3):341–346

    CAS  PubMed  Google Scholar 

  • Martin G, Juchault P, Legrand JJ (1973) Mise en évidence d’un micro-organisme intracytoplasmique symbiote de l’oniscoïde Armadillidium vulgare Latr., dont la présence accompagne l’intersexualité ou la féminisation totale des mâles génétiques de la lignée thélygène. C R Acad Sci Paris 276:2313–2316

    Google Scholar 

  • Martin G, Maissiat R, Juchault P, Legrand JJ (1974) Mise en évidence d’un micro-organisme intracytoplasmique symbiotique chez les intersexués (mâles à oostégites) du crustacé Ligia oceanica L. (Isopode, Oniscoïde). C R Acad Sci Paris 278(Série D):3375–3378

    Google Scholar 

  • Martin G, Juchault P, Sorokine O, Van Dorsselaer A (1990) Purification and characterization of androgenic hormone from the terrestrial isopod Armadillidium vulgare Latr. (Crustacea, Oniscoidea). Gen Comp Endocrinol 80:349–354

    Article  CAS  PubMed  Google Scholar 

  • Martin G, Gruppe SG, Laulier M, Bouchon D, Rigaud T, Juchault P (1994) Evidence for Wolbachia spp. in the estuarine isopod Sphaeroma rugicauda (Crustacea): a likely cytoplasmic sex ratio distorter. Endocytobiosis Cell Res 10:215–225

    Google Scholar 

  • Masui S, Sasaki T, Ishikawa H (2000) Genes for the Type IV secretion system in an intracellular symbiont, Wolbachia, a causative agent of various sexual alterations in arthropods. J Bacteriol 182(22):6529–6531

    Article  CAS  PubMed  Google Scholar 

  • Masui S, Kuroiwa H, Sasaki T, Inui M, Kuroiwa T, Ishikawa H (2001) Bacteriophage WO and virus-like particles in Wolbachia, an endosymbiont of Arthropods. Biochem Biophys Res Commun 283:1099–1104

    Article  CAS  PubMed  Google Scholar 

  • Mc Graw EA, O’Neil SL (2004) Wolbachia pipientis: intracellular infection and pathogenesis in Drosophila. Curr Opin Microbiol 7:1–4

    Article  Google Scholar 

  • Medini-Bouaziz L (2002) Systématique, biologie et biogéographie du Genre Porcellio en Tunisie (Crustacés, Isopodes Oniscoidea). Thèse de doctorat, Université El Manar, Faculté des Sciences de Tunis

  • Michel-Salzat A, Cordaux R, Bouchon D (2001) Wolbachia diversity in the Porcellionides pruinosus complex of species (Crustacea: Oniscoidea): evidence for host-dependent patterns of infection. Heredity 87:428–434

    Article  CAS  PubMed  Google Scholar 

  • Moreau J, Bertin A, Caubet Y, Rigaud T (2001) Sexual selection in an isopod with Wolbachia-induced sex reversal: males prefer real females. J Evol Biol 14:388–394

    Article  Google Scholar 

  • Moret Y, Juchault P, Rigaud T (2001) Wolbachia endosymbiont responsible for cytoplasmic incompatibility in a terrestrial crustacean: effects in natural and foreign hosts. Heredity 86:325–332

    Article  CAS  PubMed  Google Scholar 

  • Narita S, Kageyama D, Nomura M, Fukatsu T (2007) Unexpected mechanism of symbiont-induced reversel of insect sex: feminizing Wolbachia continuously acts on the butterfly Eurema hecabe during larval development. Appl Environ Microbiol 73(13):4332–4341

    Article  CAS  PubMed  Google Scholar 

  • Negri I, Pellecchia M, Mazzoglio PJ, Patetta A, Alma A (2006) Feminizing Wolbachia in Zyginidia pullula (Insecta, Hemiptera), a leafhopper with an XX/XO sex-determination system. Proc R Soc B 273:2409–2416

    Article  CAS  PubMed  Google Scholar 

  • Rigaud T, Moreau J (2004) A cost of Wolbachia-induced sex reversal and female-biased sex ratios: decrease in female fertility after sperm depletion in a terrestrial isopod. Proc R Soc Lond B 271:1941–1946

    Article  Google Scholar 

  • Rigaud T, Moreau J, Juchault P (1999) Wolbachia infection in the terrestrial isopod Oniscus asellus: sex ratio distortion and effect on fecundity. Heredity 83:469–475

    Article  PubMed  Google Scholar 

  • Rousset F, Bouchon D, Pintureau B, Juchault P, Solignac M (1992) Wolbachia endosymbionts responsible for various alterations of sexuality in arthropods. Proc R Lond B 250:91–98

    Article  CAS  Google Scholar 

  • Saitou RR, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 44:406–425

    Google Scholar 

  • Sanger F, Nicklen S, Coulson A (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Article  CAS  PubMed  Google Scholar 

  • Sun LV, Foster JM, Tzertzinis G, Ono M, Bandi C, Slatko BE, O’Neill SL (2001) Determination of Wolbachia genome size by pulsed-field gel electrophoresis. J Bacteriol 183(7):2219–2225

    Article  CAS  PubMed  Google Scholar 

  • Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The ClustalX 1 windows interface: flexible strategies for multiple sequence alignment aided by quality 2 analysis tools. Nucleic Acids Res 25:4876–4882

    Article  CAS  PubMed  Google Scholar 

  • Vandekerckhove-Tom TM, Watteyne S, Bonne W, Vanacker D, Devaere S, Rumes B, Maelfait JP, Gillis M, Swings JG, Braig HR, Mertens J (2003) Evolutionary trends in feminization and intersexuality in woodlice (Crustacean, Isopoda) infected with Wolbachia pipientis (α-proteobacteria). Belg J Zool 133(1):61–69

    Google Scholar 

  • Van de Peer Y, De Wachter R (1994) TREECON for Windows: a software package for the construction and drawing of evolutionary trees for the Microsoft Windows environment. Comput Appl Biosci 10:569–570

    PubMed  Google Scholar 

  • Verne S, Puillandre N, Brunet G, Gouin N, Samollow PB, Anderson JD, Grandjean F (2006) Characterization of polymorphic microsatellite loci in the terrestrial Isopod Armadillidium vulgare. Mol Ecol Notes 6:328–330

    Article  CAS  Google Scholar 

  • Werren JH (1997) Wolbachia run amok. Proc Natl Acad Sci USA 94:11154–11155

    Article  CAS  PubMed  Google Scholar 

  • Werren JH, Zhang W, Guo-Li R (1995) Evolution and phylogeny of Wolbachia: reproductive parasites of arthropods. Proc R Lond 261:55–71

    Article  CAS  Google Scholar 

  • Wu M, Sun LV, Vamathevan J, Riegler M, Deboy R, Brownlie JC, Mcgraw EA, Martin W, Esser C, Ahmadinejad N, Wiegand C, Madupu R, Beanan MJ, Brinkac LM, Daugherty SC, Durkin AS, Kolonay JF, Nelson WC, Mohamoud Y, Lee P, Berry K, Young MB, Utterback T, Weidman J, Nierman WC, Paulsen IT, Nelson KE, Tettelin H, O’neill SL, Eisen JA (2004) Phylogenomics of the reproductive parasite Wolbachia pipientis wMel: a streamlined genome overrun by mobile genetic elements. PloS Biol 2:327–341

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Mr. Abdellatif Boudabous, Professor and Head of the Active Microorganisms and Bimolecular Laboratory of the Scientific Faculty of Tunis, for accommodating us in his laboratory; Hayet Ketfi, Faten Godbane, Chadlya Hamdi for their valuable assistance and Kouakou Anatole for his collaboration.

Author information

Authors and Affiliations

  1. Laboratory of Reproduction and Development, Faculty of Sciences, University Tunis El Manar, 2092 El Manar II, Tunis, Tunisia

    Sarra Ben Nasr & Atf Azzouna

  2. Laboratory of Microorganisms and Actives Biomolecules, Faculty of Sciences, University Tunis El Manar, 2092 El Manar II, Tunis, Tunisia

    Maher Gtari

Authors
  1. Sarra Ben Nasr
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maher Gtari
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Atf Azzouna
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sarra Ben Nasr.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ben Nasr, S., Gtari, M. & Azzouna, A. Detection and phylogeny of the bacteria Wolbachia in the terrestrial isopod Porcellio laevis Latr. (Crustacea, Oniscoidea), isolated from Lebna and Bizerte stations, Tunisia. Ann Microbiol 60, 43–50 (2010). https://doi.org/10.1007/s13213-009-0001-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13213-009-0001-z

Keywords

Search

Navigation