Gut wall bacteria of earthworms: a natural selection process

Earthworms and microorganisms are interdependent and their interactions regulate the biogeochemistry of terrestrial soils. Investigating earthworm–microorganism interactions, we tested the hypothesis that differences in burrowing and feeding habits of anecic and endogeic earthworms are reflected by the existence of ecological group-specific gut wall bacterial communities. Bacterial community was detected using automated ribosomal intergenic spacer analysis of 16S and 23S genes and ribotype data was used to assess diversity and community composition. Using soil and earthworm samples collected from adjacent wheat–barley and grass–clover fields, we found that the anecic Lumbricus terrestris and L. friendi, the endogeic Aporrectodea caliginosa and A. longa (classically defined as anecic, but now known to possess endogeic characteristics) contain ecological group-specific gut wall-associated bacterial communities. The abundance of specific gut wall-associated bacteria (identified by sequence analysis of ribotype bands), including Proteobacteria, Firmicutes and an actinobacterium, was ecological group dependent. A microcosm study, conducted using A. caliginosa and L. terrestris and five different feeding regimes, indicated that food resource can cause shifts in gut wall-associated bacterial community, but the magnitude of these shifts did not obscure the delineation between ecological group specificity. Using A. caliginosa and A. longa samples collected in six different arable fields, we deduced that, within an ecological group, habitat was a more important determinant of gut wall-associated bacterial community composition than was host species. Hence, we conclude that the selection of bacteria associated with the gut wall of earthworms is a natural selection process and the strongest determinant of this process is in the order ecological group>habitat>species.

[1]  Francis E. Clark,et al.  Soil Organisms as Components of Ecosystems , 1978 .

[2]  N. Moran,et al.  Molecular Interactions between Bacterial Symbionts and Their Hosts , 2006, Cell.

[3]  J. Kruskal Multidimensional scaling by optimizing goodness of fit to a nonmetric hypothesis , 1964 .

[4]  O. Schmidt,et al.  Field preservation and DNA extraction methods for intestinal microbial diversity analysis in earthworms. , 2009, Journal of microbiological methods.

[5]  Thomas Dandekar,et al.  Metabolic Interdependence of Obligate Intracellular Bacteria and Their Insect Hosts , 2004, Microbiology and Molecular Biology Reviews.

[6]  N. Dubilier,et al.  Symbiotic diversity in marine animals: the art of harnessing chemosynthesis , 2008, Nature Reviews Microbiology.

[7]  T. G. Piearce,et al.  Earthworms, Their Ecology and Relationships with Soils and Land Use. , 1987 .

[8]  E. Odum Fundamentals of ecology , 1972 .

[9]  S. Marhan,et al.  Molecular profiling of 16S rRNA genes reveals diet-related differences of microbial communities in soil, gut, and casts of Lumbricus terrestris L. (Oligochaeta: Lumbricidae). , 2004, FEMS microbiology ecology.

[10]  B. Doube,et al.  Functional Interactions between Earthworms, Microorganisms, Organic Matter, and Plants , 2004 .

[11]  J. Whalen,et al.  Changes in the fatty acid profiles through the digestive tract of the earthworm Lumbricus terrestris L. , 2007 .

[12]  C. Lattaud,et al.  The diversity of digestive systems in tropical geophagous earthworms , 1998 .

[13]  U. Lohm,et al.  Soil organisms as components of ecosystem , 1977 .

[14]  K. R. Clarke,et al.  Non‐parametric multivariate analyses of changes in community structure , 1993 .

[15]  P. Lavelle,et al.  Soil Ecology , 2001, Springer Netherlands.

[16]  M. Garnett,et al.  Earthworm ecological groupings based on 14C analysis , 2005 .

[17]  G. Romanes The Formation of Vegetable Mould through the Action of Worms, with Observations on their Habits , 1881, Nature.

[18]  H. Mcculloch,et al.  THE NEW CELL PROLIFERANT , 1912 .

[19]  C. Rios-Velazquez,et al.  Characterization of possible symbionts in Onychochaeta borincana (Annelida: Glossoscolecidae) , 2007 .

[20]  N. Dubilier,et al.  Acidovorax-like symbionts in the nephridia of earthworms. , 2003, Environmental microbiology.

[21]  J. Lawton,et al.  Organisms as ecosystem engineers , 1994 .

[22]  J. Brockwell,et al.  The natural abundance of , 1985 .

[23]  C. Sorlini,et al.  Comparison of Different Primer Sets for Use in Automated Ribosomal Intergenic Spacer Analysis of Complex Bacterial Communities , 2004, Applied and Environmental Microbiology.

[24]  O. Schmidt,et al.  Stable isotope techniques in studies of the ecological diversity and functions of earthworm communities in agricultural soils , 2004 .

[25]  G. Brown,et al.  Regulation of soil organic matter dynamics and microbial activityin the drilosphere and the role of interactionswith other edaphic functional domains , 2000 .

[26]  M. Nuti,et al.  Nitrogen fixation in the gastro-enteric cavity of soil animals , 1977 .

[27]  Jesús I. Brionesa,et al.  Earthworm ecological groupings based on 14 C analysis Marı́a , 2005 .

[28]  C. D. Clegg,et al.  Spatial structure in soil chemical and microbiological properties in an upland grassland. , 2004, FEMS microbiology ecology.

[29]  O. Schmidt,et al.  Importance of DNA quality in comparative soil microbial community structure analyses , 2008 .

[30]  M. Wong,et al.  Effects of earthworm activity and P-solubilizing bacteria on P availability in soil , 2004 .

[31]  R. Knight,et al.  Evolution of Mammals and Their Gut Microbes , 2008, Science.

[32]  R. B. Misra,et al.  Endosulfan degradation by a Rhodococcus strain isolated from earthworm gut. , 2006, Ecotoxicology and environmental safety.

[33]  W. Whitman,et al.  Identification of uncultured bacteria tightly associated with the intestine of the earthworm Lumbricus rubellus (Lumbricidae; Oligochaeta) , 2003 .

[34]  S. Giovannoni,et al.  Geobacter metallireducens gen. nov. sp. nov., a microorganism capable of coupling the complete oxidation of organic compounds to the reduction of iron and other metals , 2004, Archives of Microbiology.

[35]  D. Stahl,et al.  Transmission of Nephridial Bacteria of the Earthworm Eisenia fetida , 2006, Applied and Environmental Microbiology.

[36]  J. Prosser,et al.  5 – THE PROKARYOTES , 2007 .

[37]  E. Odum Fundamentals of Ecology. , 1955 .

[38]  C. D. Clegg,et al.  Scanning electron microscopy of the gut microflora of two earthworms: Lumbricus terrestris and Octolasion cyaneum , 1993, Microbial Ecology.

[39]  P. Svastova,et al.  Earthworms (Oligochaeta, Lumbricidae) and mycobacteria. , 2003, Veterinary microbiology.

[40]  C. Scrimgeour,et al.  Dual stable isotope analysis (δ13C and δ15N) of soil invertebrates and their food sources , 2004 .

[41]  H. Drake,et al.  As the worm turns: the earthworm gut as a transient habitat for soil microbial biomes. , 2007, Annual review of microbiology.

[42]  H. Insam,et al.  Application of denaturing gradient gel electrophoresis for analysing the gut microflora of Lumbricus rubellus Hoffmeister under different feeding conditions , 2008, Bulletin of Entomological Research.

[43]  Charles Darwin,et al.  The Formation of Vegetable Mould Through the Action of Worms with Observations on Their Habits , 1881 .

[44]  A. Hastings,et al.  Using ecosystem engineers to restore ecological systems. , 2006, Trends in ecology & evolution.

[45]  C. Scrimgeour,et al.  Natural abundance of 15N and 13C in earthworms from a wheat and a wheat-clover field , 1997 .

[46]  P. Lavelle,et al.  Effects of earthworms on soil organic matter and nutrient dynamics at a landscape scale over decades , 2004 .

[47]  J. Falkinham,et al.  Humic and fulvic acids stimulate the growth of Mycobacterium avium. , 1999, FEMS microbiology ecology.

[48]  S. Marhan,et al.  Assessment of anecic behavior in selected earthworm species : Effects on wheat seed burial, seedling establishment, wheat growth and litter incorporation , 2008 .

[49]  D. Stahl,et al.  Selective recruitment of bacteria during embryogenesis of an earthworm , 2008, The ISME Journal.

[50]  Diet-related composition of the gut microbiota of Lumbricus rubellus as revealed by a molecular fingerprinting technique and cloning , 2009 .

[51]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[52]  A. Tunlid,et al.  Diversity of bacteria associated with grassland soil nematodes of different feeding groups. , 2009, FEMS microbiology ecology.

[53]  J. Amador Earthworm Ecology, 2nd ed , 2005 .

[54]  O. Giere,et al.  Microbial symbiosis in Annelida , 2005 .

[55]  A. Schramm,et al.  N2O-Producing Microorganisms in the Gut of the Earthworm Aporrectodea caliginosa Are Indicative of Ingested Soil Bacteria , 2003, Applied and Environmental Microbiology.