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Light and electron microscopic study of the bacterial adhesion to termite flagellates applying lectin cytochemistry

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Summary

Many of the flagellates inhabiting the hindgut of lower termites are associated with ectobiotic, rod-like bacteria or spirochetes. Different types of attachment sites are present. Electron dense material underlies, e.g., the plasma membrane ofJoenia annectens at the contact site, whereas other attachment sites do not show any visible specializations. The host cell's glycocalyx may, however, be reduced at the attachment sites as it is the case inDevescovina glabra. The thick glycocalyx ofStephanonympha nelumbium is not changed at the sites where bacterial rods attach, but spirochetes penetrate to a certain extent. Bacteria which colonize the extracellular surface structures ofMicrorhopalodina multinucleata express their own glycocalyx to mediate a contact. In this study we focussed on the examination of one common mode of interaction between bacteria and their host cells, i.e., adhesion via lectins and sugars. The sugar composition was analysed by light and electron microscopic labelling experiments using the lectins Con A, WGA and SBA. In general, only the posterior body surface ofJoenia which is colonized with bacteria is labelled. The demonstrated sugars are found in fibrous glycocalyx portions surrounding the attachment sites of the bacteria. Such glycocalyx fibres in combination with the electron dense material supporting the attachment sites seem to be the prerequisites for bacterial attachment. InD. glabra, however, a role for sugars in mediating the attachment could not be demonstrated. Removal of the ectobiotes using antibiotics revealed that the specialized contact sites ofJoenia are present in the absence of bacteria and thus possibly serve to attract bacteria. Nothing, however, remains of the former attachment sites in bacteria-freeDevescovina cells. Attachment sites in this case could be induced by bacterial contact. There is not one general mechanism for bacterial attachment to termite flagellates; rather, adhesion seems to follow different strategies.

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Abbreviations

Con:

A concanavalin A

DAB:

3,3′-diaminobenzidine tetrahydrochloride

DAPI:

4′,6-diamidino-2-phenylindole

DIC:

differential interference contrast

FA:

formaldehyde

FITC:

fluorescein isothiocyanate

GA:

glutaraldehyde

PB:

Soerensen's phosphate buffer

PC:

phase contrast

pen/strep:

penicillin and streptomycin

SBA:

soybean agglutinin

SEM:

scanning electron microscope

TBS:

Tris buffer saline

TEM:

transmission electron microscope

WGA:

wheat germ agglutinin

References

  • Bernhard W, Avrameas S (1971) Ultrastructural visualization of cellular carbohydrate components by means of concanavalin A. Exp Cell Res 64: 232–236

    PubMed  Google Scholar 

  • Bloodgood RA, Fitzharris TP (1976) Specific association of prokaryotes with symbiotic flagellate protozoa from the hindgut of the termiteReticulitermes and the wood-eating roachCryptocercus. Cytobios 17: 103–122

    PubMed  Google Scholar 

  • Cardullo RA, Wolf DE (1990) The sperm plasma membrane: a little more than mosaic, a little less than fluid. In: Bloodgood RA (ed) Ciliary and flagellar membranes. Plenum, New York, pp 305–336

    Google Scholar 

  • Carlemalm E, Garavito RM, Villinger W (1982) Resin development for electron microscopy and an analysis of embedding at low temperature. J Microsc 126: 123–143

    Google Scholar 

  • Christensen GD, Simpson WA, Beachey EH (1985) Adhesion of bacteria to animal tissues. Complex mechanisms. In: Savage DC, Fletcher M (eds) Bacterial adhesion. Plenum, New York, pp 279–305

    Google Scholar 

  • Cleveland LR (1951) Hormone-induced sexual cycles of flagellates. VII. One-division meiosis and autogamy without cell division inUrinympha. J Morphol 88: 385–439

    Google Scholar 

  • —, Grimstone AV (1964) The fine structure of the flagellateMlxotricha paradoxa and its associated microorganisms. Proc R Soc Lond Biol 159: 668–686

    Google Scholar 

  • Costerton JW, Geesey GG, Chen K-J (1978) How bacteria stick. Sci Am 238: 86–95

    PubMed  Google Scholar 

  • Costerton JW, Irvin RT, Cheng K-J (1981) The bacterial glycocalyx in nature and disease. Annu Rev Microbiol 35: 299–324

    PubMed  Google Scholar 

  • Doyle RJ (1991) Strategies in experimental microbial adhesion research. In: Mozes N, Handley PS, Busscher HJ, Rouxhet PG (eds) Microbial cell surface analysis: structural and physico-chemical methods. VCH, Weinheim, pp 291–316

    Google Scholar 

  • Dyer BD, Khalsa O (1993) Surface bacteria ofStreblomastix strix are sensory symbionts. BioSystems 31: 169–180

    PubMed  Google Scholar 

  • Gabius H-J, Gabius S, Zemlyanukhina TV, Bovin NV, Brinck U, Danguy A, Joshi SS, Kayser K, Schottelius J, Sinowatz F, Tietze LF, Vidal-Vanaclocha F, Zanetta J-P (1993) Reverse lectin histochemistry: design and application of glycoligands for detection of cell and tissue lectins. Histol Histopathol 8: 369–383

    PubMed  Google Scholar 

  • Goldstein U, Poretz RD (1986) Isolation, physicochemical characterization, and carbohydrate-binding specificity of lectins. In: Liener IE. Sharon N, Goldstein IJ (eds) The lectins: properties, functions, and applications in biology and medicine. Academic Press, Orlando, pp 35–250

    Google Scholar 

  • Graham RC, Karnovsky MJ (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14: 291–302

    PubMed  Google Scholar 

  • Gumbiner B, Louvard D (1985) Localized barriers in the plasma membrane: a common way to form domains. Trends Biochem Sci 10: 435–438

    Google Scholar 

  • Hancock IC (1991) Microbial cell surface architecture. In: Mozes N, Handley PS, Busscher HJ, Rouxhet PG (eds) Microbial cell surface analysis: structural and physicochemical methods. VCH, Weinheim, pp 21–59

    Google Scholar 

  • Hollande A (1979) Complexes parabasaux et symétrie bilatérale du rostre chezJoenia annectens. Protistologica 15: 407–426

    Google Scholar 

  • —, Valentin J (1969) Appareil de Golgi, pinocytose, lysosomes, mitochondries, bactéries symbiontiques, atractophores et pleuromitose chez les Hypermastigines du genreJoenia. Affinités entre Joeniides et Trichomonadines. Protistologica 5: 39–86

    Google Scholar 

  • Horisberger M (1984) Lectin cytochemistry. In: Polak JM, Varndell IM (eds) Immunolabelling for electron microscopy. Elsevier, Amsterdam, pp 249–258

    Google Scholar 

  • Kirby H (1928) A species ofProboscidiella fromKalotermes (Cryptotermes) dudleyi Banks, a termite of Central America, with remarks on the oxymonad flagellates. Q J Microsc Sci 72: 355–386

    Google Scholar 

  • — (1941) Devescovinid flagellates of termites. 1. The genusDevescovina. Univ Calif Publ Zool 45: 1–91

    Google Scholar 

  • —, Margulis L (1994) Harold Kirby's symbionts of termites: karyomastigont reproduction and calonymphid taxonomy. Symbiosis 16: 7–63

    PubMed  Google Scholar 

  • Lavette A (1977) L'appareil parabasal deJoenia annectens (Zooflagellata Joeniidea) est un organite composite. C R Acad Sci Ser D 284: 449–452

    Google Scholar 

  • Matthysse AG (1992) Bacterial adhesion. In: Lederberg J (ed) Encyclopedia of microbiology, vol 1. Academic Press, London, pp 29–36

    Google Scholar 

  • Nicolson GL (1974) The interactions of lectins within animal surfaces. Int Rev Cytol 39: 89–183

    PubMed  Google Scholar 

  • Nilsson J (1989)Tetrahymena in cytotoxicology: with special reference to effects of heavy metals and selected drugs. Eur J Protistol 25: 2–25

    Google Scholar 

  • Ofek I, Doyle RJ (eds) (1994) Bacterial adhesion to cells and tissues. Chapman and Hall, New York

    Google Scholar 

  • Paulin JJ (1992) Ruthenium red as a stain. In: Lee JJ, Soldo AT (eds) Protocols in protozoology. Allen Press, Lawrence, C 21.1

    Google Scholar 

  • Radek R, Hausmann K, Breunig A (1992) Ectobiotic and endocytobiotic bacteria associated with the termite flagellateJoenia annectens. Acta Protozool 31: 93–107

    Google Scholar 

  • Reeke GN Jr, Becker JW (1988) Carbohydrate-binding sites of plant lectins. Curr Top Microbiol Immunol 139: 35–58

    PubMed  Google Scholar 

  • Reynolds ES (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol 17: 208–212

    PubMed  Google Scholar 

  • Rosenberg M, Kjelleberg S (1986) Hydrophobie interactions: role in bacterial adhesion. In: Marshall KC (ed) Advances in microbial ecology, vol 9. Plenum, New York, pp 353–393

    Google Scholar 

  • Roth J, Lucocq JM, Charest PM (1984) Light and electron microscopic demonstration of sialic acid residues with the lectin fromLimax flavus. J Histochem Cytochem 32: 1167–1176

    PubMed  Google Scholar 

  • Schachter J (1988) The intracellular life ofChlamydia. Curr Top Microbiol Immunol 138: 109–140

    PubMed  Google Scholar 

  • Shaklai M, Tavassoli M (1982) Lanthanum as an electron microscopic stain. J Histochem Cytochem 30: 1325–1330

    PubMed  Google Scholar 

  • Sharon N (1986) Bacterial lectins. In: Liener IE, Sharon N, Goldstein IJ (eds) The lectins: properties, functions, and applications in biology and medicine. Academic Press, London, pp 494–526

    Google Scholar 

  • Slifkin M, Doyle RJ (1990) Lectins and their application to clinical microbiology. Clin Microbiol Res 3: 197–218

    Google Scholar 

  • Spurr AR (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26: 31–43

    PubMed  Google Scholar 

  • Strömberg N, Hultgren S, Russell DG, Normark S (1992) Microbial attachment, molecular mechanisms. In: Lederberg J (ed) Encyclopedia of microbiology, vol 3. Academic Press, London, pp 143–158

    Google Scholar 

  • Taatjes DJ, Schaub U, Roth J (1987) Light microscopical detection of antigens and lectin binding sites with gold-labelled reagents on semi-thin Lowicryl K4M sections: usefulness of the photochemical silver reaction for signal amplification. Histochem J 19: 235–245

    PubMed  Google Scholar 

  • Tamm SL (1980) The ultrastructure of prokaryotic-eukaryotic cell junctions. J Cell Sci 44: 335–352

    PubMed  Google Scholar 

  • — (1982) Flagellated ectosymbiotic bacteria propel a eukaryotic cell. J Cell Biol 94: 697–709

    PubMed  Google Scholar 

  • Yamin MA (1981) Cellulose metabolism by the flagellateTrichonympha from a termite is independent of endosymbiotic bacteria. Science 211: 58–59

    Google Scholar 

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Authors and Affiliations

  1. Department of Protozoology, Institute of Zoology, Free University of Berlin, Königin-Luise-Strasse 1-3, D-14195, Berlin, Federal Republic of Germany

    Renate Radek, Joachim Rösel & Klaus Hausmann

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  1. Renate Radek
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  2. Joachim Rösel
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  3. Klaus Hausmann
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Dedicated to Prof. Dr. Dr. h.c. Eberhard Schnepf on the occasion of his retirement

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Radek, R., Rösel, J. & Hausmann, K. Light and electron microscopic study of the bacterial adhesion to termite flagellates applying lectin cytochemistry. Protoplasma 193, 105–122 (1996). https://doi.org/10.1007/BF01276639

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  • DOI: https://doi.org/10.1007/BF01276639

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