Survival of earthworms in the environment depends on their ability to recognize and eliminate potential pathogens. This work is aimed to compare the innate defense mechanisms of two closely related earthworm species, Eisenia andrei and Eisenia fetida, that inhabit substantially different ecological niches. While E. andrei lives in a compost and manure, E. fetida can be found in the litter layer in forests. Therefore, the influence of environment-specific microbiota on the immune response of both species was followed. Firstly, a reliable method to discern between E. andrei and E. fetida based on species-specific primers for cytochrome c oxidase I (COI) and stringent PCR conditions was developed. Secondly, to analyze the immunological profile in both earthworm species, the activity and expression of lysozyme, pattern recognition protein CCF, and antimicrobial proteins with hemolytic function, fetidin and lysenins, have been assessed. Whereas, CCF and lysozyme showed only slight differences in the expression and activity, fetidin/lysenins expression as well as the hemolytic activity was considerably higher in E. andrei as compared to E. fetida. The expression of fetidin/lysenins in E. fetida was not affected upon the challenge with compost microbiota, suggesting more substantial changes in the regulation of the gene expression. Genomic DNA analyses revealed significantly higher level of fetidin/lysenins (determined using universal primer pairs) in E. andrei compared to E. fetida. It can be hypothesized that E. andrei colonizing compost as a new habitat acquired an evolutionary selection advantage resulting in a higher expression of antimicrobial proteins.

Laboratory of Cellular and Molecular Immunology Institute of Microbiology of the Academy of Sciences of the Czech Republic v v i Prague Czech Republic

Zobrazit více v PubMed

Cooper EL (1968) Transplantation immunity in annelids. I. Rejection of xenografts exchanged between Lumbricus terrestris and Eisenia foetida . Transplantation 6: 322-327. doi:10.1097/00007890-196805000-00002. PubMed: 4871392. PubMed DOI

Bailey S, Miller BJ, Cooper EL (1971) Transplantation immunity in annelids. II. Adoptive transfer of the xenograft reaction. Immunology 21: 81-86. PubMed: 5558033. PubMed PMC

Jollès P (1996) From the discovery of lysozyme to the characterization of several lysozyme families. EXS 75: 3-5. PubMed: 8765291. PubMed

Josková R, Silerová M, Procházková P, Bilej M (2009) Identification and cloning of an invertebrate-type lysozyme from Eisenia andrei . Dev Comp Immunol 33: 932-938. doi:10.1016/j.dci.2009.03.002. PubMed: 19454335. PubMed DOI

Roch P, Valembois P, Davant N, Lassegues M (1981) Protein analysis of earthworm coelomic fluid. II. Isolation and biochemical characterization of the Eisenia foetida andrei factor (EFAF). Comp Biochem Physiol 69B: 829-836.

Milochau A, Lassègues M, Valembois P (1997) Purification, characterization and activities of two hemolytic and antibacterial proteins from coelomic fluid of the annelid Eisenia fetida andrei . Biochim Biophys Acta 1337: 123-132. doi:10.1016/S0167-4838(96)00160-4. PubMed: 9003444. PubMed DOI

Sekizawa Y, Kubo T, Kobayashi H, Nakajima T, Natori S (1997) Molecular cloning of cDNA for lysenin, a novel protein in the earthworm Eisenia foetida that causes contraction of rat vascular smooth muscle. Gene 191: 97-102. doi:10.1016/S0378-1119(97)00047-4. PubMed: 9210594. PubMed DOI

Bilej M, Brys L, Beschin A, Lucas R, Vercauteren E et al. (1995) Identification of a cytolytic protein in the coelomic fluid of Eisenia foetida earthworms. Immunol Lett 45: 123-128. doi:10.1016/0165-2478(94)00248-P. PubMed: 7622179. PubMed DOI

Beschin A, Bilej M, Torreele E, De Baetselier P (2001) On the existence of cytokines in invertebrates. Cell Mol Life Sci 58: 801-814. doi:10.1007/PL00000901. PubMed: 11437239. PubMed DOI PMC

Köhlerová P, Beschin A, Silerová M, De Baetselier P, Bilej M (2004) Effect of experimental microbial challenge on the expression of defense molecules in Eisenia foetida earthworm. Dev Comp Immunol 28: 701-711. doi:10.1016/j.dci.2004.01.001. PubMed: 15043940. PubMed DOI

Silerová M, Procházková P, Josková R, Josens G, Beschin A et al. (2006) Comparative study of the CCF-like pattern recognition protein in different Lumbricid species. Dev Comp Immunol 30: 765-771. doi:10.1016/j.dci.2005.11.002. PubMed: 16386303. PubMed DOI

Bouché MB (1972) Lombriciens de France. Écologie et Systématique. I N R A Publ Ann Zool Ecol Anim 72.

André F (1963) Contribution a l´analyse experimental de la reproduction des lombriciens. Bull Biol Fr Belg 97: 1-101.

Dominguez J, Velando A, Ferreiro A (2005) Are Eisenia fetida (Savigny, 1826) and Eisenia andrei Bouche (1972) (Oligochaeta, Lumbricidae) different biological species? Pedobiologia 49.

Perez-Losada M, Eiroa J, Mato S, Dominguez J (2005) Phylogenetic species delimitation of the earthworms Eisenia fetida (Savigny, 1826) and Eisenia andrei Bouche, 1972 (Oligochaeta, Lumbricidae) based on mitochondrial and nuclear DNA sequences. Pedobiologia 49: 317-324. doi:10.1016/j.pedobi.2005.02.004. DOI

Pižl V (2002) Earthworms of the Czech Republic. Sborník Přírodovědného klubu v U. Hradišti (in Czech) Suppl. 9: 1-154

Sanger F, Nicklen S, Coulson AR (1977) DNA Sequencing with Chain-Terminating Inhibitors. Proc Natl Acad Sci U S A 74: 5463-5467. doi:10.1073/pnas.74.12.5463. PubMed: 271968. PubMed DOI PMC

Murray V (1989) Improved Double-Stranded DNA Sequencing Using the Linear Polymerase Chain-Reaction. Nucleic Acids Res 17: 8889-8889. doi:10.1093/nar/17.21.8889. PubMed: 2587244. PubMed DOI PMC

Lie O, Syed M, Solbu H (1986) Improved agar plate assays of bovine lysozyme and haemolytic complement activity. Acta Vet Scand 27: 23-32. PubMed: 3751812. PubMed PMC

Kyselková M, Chroňáková A, Volná L, Nĕmec J, Ulmann V et al. (2012) Tetracycline resistance and presence of tetracycline resistance determinants tet(V) and tap in rapidly growing mycobacteria from agricultural soils and clinical isolates. Microbes Environ 27: 413-422. doi:10.1264/jsme2.ME12028. PubMed: 22673307. PubMed DOI PMC

Bruce KD, Hiorns WD, Hobman JL, Osborn AM, Strike P et al. (1992) Amplification of DNA from native populations of soil bacteria by using the polymerase chain reaction. Appl Environ Microbiol 58: 3413-3416. PubMed: 1444376. PubMed PMC

Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41: 95-98.

Chun J, Lee JH, Jung Y, Kim M, Kim S et al. (2007) EzTaxon: a web-based tool for the identification of prokaryotes based on 16S ribosomal RNA gene sequences. Int J Syst Evol Microbiol 57: 2259-2261. doi:10.1099/ijs.0.64915-0. PubMed: 17911292. PubMed DOI

Cotuk A, Dales RP (1984) Lysozyme activity in the coelomic fluid and coelomocytes of the earthworm Eisenia foetida SAV in relation to bacterial infection. Comp Biochem Physiol 78A: 469-474.

Beschin A, Bilej M, Hanssens F, Raymakers J, Van Dyck E et al. (1998) Identification and cloning of a glucan- and lipopolysaccharide-binding protein from Eisenia foetida earthworm involved in the activation of prophenoloxidase cascade. J Biol Chem 273: 24948-24954. doi:10.1074/jbc.273.38.24948. PubMed: 9733802. PubMed DOI

Kauschke E, Pagliara P, Stabili L, Cooper EL (1997) Characterization of Proteolytic Activity in Coelomic Fluid of Lumbricus terrestris L. (Annelida, Lumbricida). Comp Biochem Physiol 116 (B): 235-242

Lavelle P (1988) Earthworm activities and the soil system. Biol Fertil Soils 6: 237-251.

Procházková P, Sustr V, Dvořák J, Roubalová R, Skanta F et al. (2013) Correlation between the activity of digestive enzymes and nonself recognition in the gut of Eisenia andrei earthworms. J Invertebr Pathol: 114: 217-221. doi:10.1016/j.jip.2013.08.003. PubMed: 23999244. PubMed DOI

Du Pasquier L, Duprat P (1968) Humoral and cellular aspects of a non-specific natural immunity in the oligochetee Eisenia foetida Sav. (Lumbricinae). C R Acad Sci Hebd Seances Acad Sci D. pp. 538-541. PubMed

Lassegues M, Milochau A, Doignon F, Du Pasquier L, Valembois P (1997) Sequence and expression of an Eisenia-fetida-derived cDNA clone that encodes the 40-kDa fetidin antibacterial protein. Eur J Biochem 246: 756-762. doi:10.1111/j.1432-1033.1997.00756.x. PubMed: 9219536. PubMed DOI

Vitturi R, Colomba MS, Pirrone A, Libertini A (2000) Physical mapping of rDNA genes, (TTAGGG)n telomeric sequence and other karyological features in two earthworms of the family Lumbricidae (Annelida: Oligochaeta). Heredity (Edinb) 85 3: 203-207. doi:10.1046/j.1365-2540.2000.00709.x. PubMed: 11012723. PubMed DOI

Gregory TR, Hebert PDN (2002) Genome size estimates for some oligochaete annelids. Can J Zool 80: 1485-1489. doi:10.1139/z02-145. DOI

Procházková P, Silerová M, Felsberg J, Josková R, Beschin A et al. (2006) Relationship between hemolytic molecules in Eisenia fetida earthworms. Dev Comp Immunol 30: 381-392. doi:10.1016/j.dci.2005.06.014. PubMed: 16051356. PubMed DOI

Valembois P, Roch P, Lassegues M (1986) Antibacterial molecules in annelids. In: Brehelin M. Immunity in Invertebrates. Berlin, Heidelberg, New York: Springer-Verlag; pp. 74-93.

Roch P, Lassegues M, Valembois P (1991) Antibacterial activity of Eisenia fetida andrei coelomic fluid: III-Relationship within the polymorphic hemolysins. Dev Comp Immunol 15: 27-32. doi:10.1016/0145-305X(91)90044-Y. PubMed: 2050244. PubMed DOI

Cross-Species Comparisons of Nanoparticle Interactions with Innate Immune Systems: A Methodological Review

The Effects of In Vivo Exposure to Copper Oxide Nanoparticles on the Gut Microbiome, Host Immunity, and Susceptibility to a Bacterial Infection in Earthworms

Developmental and Immune Role of a Novel Multiple Cysteine Cluster TLR From Eisenia andrei Earthworms

Contribution of Eisenia andrei earthworms in pathogen reduction during vermicomposting

Zobrazit více v PubMed

GENBANK
HQ534065, HQ534066