Advertisement

Marine Biology

, Volume 148, Issue 3, pp 631–642 | Cite as

Characterizing the resident, fermentative microbial consortium in the hindgut of the temperate-zone herbivorous fish, Hermosilla azurea (Teleostei: Kyphosidae)

  • Pat M. Fidopiastis
  • Daniel J. Bezdek
  • Michael H. Horn
  • Judith S. Kandel
Research Article

Abstract

The zebraperch, Hermosilla azurea Jenkins and Evermann, a warm-temperate marine fish species with a strictly macroalgal diet, has a relatively long digestive tract with an enlarged hindgut and an associated blind caecum (HC). In zebraperch sampled off Santa Catalina Island, California (33°19′42′′N; 118°18′37′′W) in years 1995 through 2001, direct cell counts, gut epithelium assessment of bacterial attachment, and short-chain fatty acid (SCFA) analyses verified that the zebraperch HC possesses a dense and morphologically diverse, fermentative microbiota. Bacterial cell counts and morphological diversity were significantly higher in HC contents compared to anterior gut regions, suggesting that microbial populations were growing along the digestive tract. Similarly, electron micrographs of the HC epithelium revealed attached microbes, further supporting the possibility that these organisms constitute resident microbiota. Five different SCFAs were detected in all three regions of the digestive tract, but levels were up to three times greater in HC contents. Acetate was consistently the prevailing SCFA in all gut regions. Sequence analysis of bacterial 16S rDNA was used to identify predominant bacterial groups in HC contents. Of the seven main bacterial types identified, Enterovibrio spp. were the dominant bacteria in HC contents followed by species of Bacteroides,Faecalibacterium, and Desulfovibrio. Taken together, our findings show that the zebraperch HC harbors a consortium of microbes that appears to assist in the breakdown of algal polysaccharides in the herbivorous diet of the fish.

Keywords

Digestive Tract Desulfovibrio Herbivorous Fish Algal Polysaccharide SCFA Level 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We thank Captain J. Cvitanovich and the crew of the R.V. Yellowfin for their help in capturing fish. We also thank W. Van Antwerp for performing SCFA analyses, E. Sturm for leading several expeditions to collect fish, and E. DeLong for his generous donation of PCR primers. T. Parker, J. Ferreira, W. Katzenstein, C. Hamilton, D. Tessier, S. Ecker, J. Haygood, and D. Asher provided valuable assistance with fish collection and lab work. The Departmental Associations Council and the Department of Biological Science at California State University, Fullerton, provided financial support. All experiments herein comply with the laws of the United States of America.

References

  1. Altschul SF, Gish W, Miller W, Meyers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410CrossRefGoogle Scholar
  2. Amann RI, Ludwig W, Schleiffer KH (1995) Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol Rev 59:143–169PubMedPubMedCentralGoogle Scholar
  3. Bernhard AE, Field KG (2000) Identification of nonpoint sources of fecal pollution in coastal waters by using host-specific 16S ribosomal DNA genetic markers from fecal anaerobes. Appl Environ Microbiol 66:1587–1594CrossRefGoogle Scholar
  4. Brune A, Friedrich MW (2000) Microecology of the termite gut: structure and function on a microscale. Curr Opin Microbiol 3:263–269CrossRefGoogle Scholar
  5. Choat JH, Clements KD (1998) Vertebrate herbivores in marine and terrestrial environments: a nutritional ecology perspective. Annu Rev Ecol Syst 29:375–403CrossRefGoogle Scholar
  6. Choat JH, Clements KD, Robbins WD (2002) The trophic status of herbivorous fishes on coral reefs. Mar Biol 140:613–623CrossRefGoogle Scholar
  7. Clements KD (1991) Endosymbiotic communities of two herbivorous labroid fishes, Odax cyanomelas and O. pullus. Mar Biol 109:223–229CrossRefGoogle Scholar
  8. Clements KD, Choat JH (1995) Fermentation in tropical marine herbivorous fishes. Physiol Zool 68:355–378CrossRefGoogle Scholar
  9. Clements KD, Choat JH (1997) A comparison of herbivory in the closely related marine fish genera Girella and Kyphosus. Mar Biol 127:579–586CrossRefGoogle Scholar
  10. Clements KD, Gleeson VP, Slaytor M (1994) Short-chain fatty acid metabolism in temperate marine herbivorous fish. J Comp Physiol B 168:61–72CrossRefGoogle Scholar
  11. DeLong EF, Franks DG, Alldredge AL (1993) Phylogenetic diversity of aggregate attached vs. free-living marine bacterial assemblages. Limnol Oceanogr 38:924–934CrossRefGoogle Scholar
  12. Deplancke B, Hristova KR, Oakley HA, McCracken VJ, Aminov R, Mackie RI, Gaskins HR (2000) Molecular ecological analysis of the succession and diversity of sulfate-reducing bacteria in the mouse gastrointestinal tract. Appl Environ Microbiol 66:2166–2174CrossRefGoogle Scholar
  13. Duncan SH, Hold GL, Harmsen HJM, Stewart CS, Flint HJ (2002) Growth requirements and fermentation products of Fusobacterium prausnitzii, and a proposal to reclassify it as Faecalibacterium prausnitzii gen. nov., comb. nov. Int J Syst Evol Microbiol 52:2141–2146PubMedGoogle Scholar
  14. Duncan SH, Holtrop G, Lobley GE, Calder AG, Stewart CS, Flint HJ (2004) Contribution of acetate to butyrate formation by human faecal bacteria. Br J Nutr 91:915–923CrossRefGoogle Scholar
  15. Fuhrman JA, Lee SH, Masuchi Y, Davis AA, Wilcox RM (1994) Characterization of marine prokaryotic communities via DNA and RNA. Microb Ecol 28:133–145CrossRefGoogle Scholar
  16. Giovannoni SJ, Britschgi TB, Moyer CL, Field KG (1990) Genetic diversity in Sargasso Sea bacterioplankton. Nature 345:60–62CrossRefGoogle Scholar
  17. Hansen GH, Olafsen JA (1999) Bacterial interactions in early life stages of marine cold water fish. Microb Ecol 38:1–26CrossRefGoogle Scholar
  18. Hobbie JE, Jasper S, Daley RJ (1977) Use of nucleopore filters for counting bacteria by fluorescence microscopy. Appl Environ Microbiol 33:1225–1228PubMedPubMedCentralGoogle Scholar
  19. Horn MH (1989) Biology of marine herbivorous fishes. Oceanogr Mar Biol Annu Rev 27:167–272Google Scholar
  20. Horn MH (1992) Herbivorous fishes: feeding and digestive mechanisms. In: John DM, Hawkins SJ, Price JH (eds) Plant–animal interactions in the marine benthos. Systematics association special, vol 46. Clarendon Press, Oxford, pp 339–362Google Scholar
  21. Kandel JS, VanAntwerp W, Horn MH (1994) Volatile fatty acids in the hindguts of herbivorous fishes from temperate and tropical marine waters. J Fish Biol 45:527–529CrossRefGoogle Scholar
  22. Kreader CA (1998) Persistence of PCR-detectable Bacteroides distasonis from human feces in river water. Appl Environ Microbiol 64:4103–4105PubMedPubMedCentralGoogle Scholar
  23. Kurita-Ochiai T, Ochiai K, Fukushima K (1998) Volatile fatty acid, metabolic by-product of periodontopathic bacteria, induces apoptosis in WEHI 231 and RAJI B lymphoma cells and splenic B cells. Infect Immun 66:2587–2594PubMedPubMedCentralGoogle Scholar
  24. Liston J (1957) The occurrence and distribution of bacterial types on flatfish. J Gen Microbiol 16:205–216CrossRefGoogle Scholar
  25. Lobel PS (1981) Trophic biology of herbivorous fishes: alimentary pH and digestive capabilities. J Fish Biol 19:365–397CrossRefGoogle Scholar
  26. Lu J, Idris U, Harmon B, Hofacre C, Maurer JJ, Lee MD (2003) Diversity and succession of the intestinal bacterial community of the maturing broiler chicken. Appl Environ Microbiol 69:6816–6824CrossRefGoogle Scholar
  27. Luczkovich JJ, Stellwag EJ (1993) Isolation of cellulolytic microbes from the intestinal tract of the pinfish, Lagodon rhomboides: size-related changes in diet and microbial abundance. Mar Biol 116:381–388CrossRefGoogle Scholar
  28. MacCormack WP, Fraile ER (1990) Bacterial flora of newly caught Antarctic fish Notothenia neglecta. Polar Bio 10:413–417Google Scholar
  29. McCandless EL, Craigie JS (1979) Sulfated polysaccharides in red and brown algae. Ann Rev Plant Physiol 30:41–53CrossRefGoogle Scholar
  30. Mountfort DO, Campbell J, Clements KD (2002) Hindgut fermentation in three species of marine herbivorous fish. Appl Environ Microbiol 68:1374–1380CrossRefGoogle Scholar
  31. Muddaris M, Austin B (1988) Quantitative and qualitative studies of the bacterial microflora of turbot, Scopthalmus maximus, gills. J Fish Biol 32:223–229CrossRefGoogle Scholar
  32. Narayanan S, Nagaraja TG, Okwumabua O, Staats J, Chengappa MM, Oberst RD (1997) Ribotyping to compare Fusobacterium necrophorum isolates from bovine liver abscesses, ruminal walls, and ruminal contents. Appl Environ Microbiol 63:4671–4678PubMedPubMedCentralGoogle Scholar
  33. Paster BJ, Dewhirst FE, Cooke SM, Fussing V, Poulsen LK, Breznak JA (1996) Phylogeny of not-yet cultured spirochetes from termite guts. Appl Environ Microbiol 62:347–352PubMedPubMedCentralGoogle Scholar
  34. Penry DL, Jumars PA (1987) Modeling animal guts as chemical reactors. Am Nat 129:69–92CrossRefGoogle Scholar
  35. Ravenschlag K, Sahm K, Pernthaler J, Amann R (1999) High bacterial diversity in permanently cold marine sediments. Appl Environ Microbiol 65:3982–3989PubMedPubMedCentralGoogle Scholar
  36. Reddy NR, Palmer JK, Pierson MD, Bothast RJ (1984) Intracellular glycosidases of human colon Bacteroides ovatus B4-11. Appl Environ Microbiol 48:890–892PubMedPubMedCentralGoogle Scholar
  37. Rimmer DW (1986) Changes in diet and the development of microbial digestion in juvenile buffalo bream, Kyphosus cornelii. Mar Biol 92:443–448CrossRefGoogle Scholar
  38. Rimmer DW, Wiebe RJ (1987) Fermentative microbial digestion in herbivorous fishes. J Fish Biol 31:229–236CrossRefGoogle Scholar
  39. Ringo E, Lødemel JB, Myklebust R, Kaino T, Mayhew TM, Olsen RE (2001) Epithelium-associated bacteria in the gastrointestinal tract of Arctic charr (Salvelinus alpinus L.). An electron microscopical study. J Appl Microbiol 90:294–300CrossRefGoogle Scholar
  40. Salyers AA, Vercellotti JR, West SEH, Wilkins TD (1977) Fermentation of mucin and plant polysaccharides by strains of Bacteroides from the human colon. Appl Environ Microbiol 33:319–322PubMedPubMedCentralGoogle Scholar
  41. Seeto GS, Veivers PC, Clements KD, Slaytor M (1996) Carbohydrate utilization by microbial symbionts in the marine herbivorous fishes Odax cyanomelas and Crinodus lophodon. J Comp Physiol B 165:571–579CrossRefGoogle Scholar
  42. Skea GL, Mountfort DO, Clements KD (2005) Gut carbohydrases from the New Zealand marine herbivorous fishes Kyphosus sydneyanus (Kyphosidae), Aplodactylus arctidens (Aplodactylidae) and Odax pullus (Labridae). Comp Biochem Physiol B 140:259–269CrossRefGoogle Scholar
  43. Sturm EA, Horn MH (1998) Food habits, gut morphology and pH, and assimilation efficiency of the zebraperch Hermosilla azurea, an herbivorous kyphosid fish of temperate marine waters. Mar Biol 132:515–522CrossRefGoogle Scholar
  44. Sturm EA, Horn MH (2001) Increase in occurrence and abundance of zebraperch (Hermosilla azurea) in the Southern California Bight in recent decades. Bull So Cal Acad Sci 100:170–174Google Scholar
  45. Suau A, Bonnet R, Sutren M, Godon J-J, Gibson GR, Collins MD, Dore J (1999) Direct analysis of genes encoding 16S rRNA from complex communities reveals many novel molecular species within the human gut. Appl Environ Microbiol 65:4799–4807PubMedPubMedCentralGoogle Scholar
  46. Sugita H, Tsunohara M, Ohkoshi T, Deguchi Y (1988) The establishment of an intestinal microflora in developing goldfish (Carassisus auratus) of culture ponds. Microb Ecol 15:333–344CrossRefGoogle Scholar
  47. Sutton D, Clements KD (1988) Aerobic heterotrophic gastrointestinal microflora of tropical marine fishes. Proc Sixth Int Coral Reef Symp (Aust) 3:185–190Google Scholar
  48. Thompson FL, Hoste B, Thompson CC, Goris J, Gomez-Gil B, Huys L, De Vos P, Swings J (2002) Enterovibrio norvegicus gen. nov., sp. nov., isolated from the gut of turbot (Scopthalmus maximus) larvae: a new member of the family Vibrionaceae. Int J Syst Evol Microbiol 52:2015–2022PubMedGoogle Scholar
  49. Thompson FL, Thompson CC, Naser S, Hoste B, Vandemeulebroecke K, Munn C, Bourne D, Swings J (2005) Photobacterium rosenbergii sp. nov. and Enterovibrio coralii sp. nov., vibrios associated with coral bleaching. Int J Syst Evol Microbiol 55:913–917CrossRefGoogle Scholar
  50. Titus E, Ahearn GA (1991) Transintestinal acetate transport in a herbivorous teleost: anion exchange at the basolateral membrane. J Exp Biol 156:41–61Google Scholar
  51. Tran CP, Familari M, Parker LM, Whitehead RH, Giraud AS (1998) Short-chain fatty acids inhibit intestinal trefoil factor gene expression in colon cancer cells. Am J Physiol Gastrointest Liver Physiol 275:G85–G94CrossRefGoogle Scholar
  52. Tsai HH, Sunderland D, Gibson GR, Hart CA, Rhodes JM (1992) A novel mucin sulphatase from human feces: its identification, purification, and characterization. Clin Sci 82:447–454CrossRefGoogle Scholar
  53. Van der Maarel MJ, Artz R, Haanstra R, Forney LJ (1998) Association of marine archaea with the digestive tracts of two marine fish species. Appl Environ Microbiol 64:2894–2898Google Scholar
  54. Wang X, Conway PL, Brown IL, Evans AJ (1999) In vitro utilization of high-amylose maize (amylomaize) starch granules by human colonic bacteria. Appl Environ Microbiol 65:4848–4854PubMedPubMedCentralGoogle Scholar
  55. Whitford MF, Forster RJ, Beard CE, Gong J, Teather RM (1998) Phylogenetic analysis of rumen bacteria by comparative sequence analysis of cloned 16S rRNA genes. Anaerobe 4:153–163CrossRefGoogle Scholar
  56. Willis CL, Cummings JH, Neale G, Gibson GR (1996) In vitro effects of mucin fermentation on the growth of human colonic sulphate-reducing bacteria. Anaerobe 2:117–122CrossRefGoogle Scholar
  57. Wilson KH, Blitchington RB (1996) Human colonic biota studied by ribosomal DNA sequence analysis. Appl Environ Microbiol 62:2273–2278PubMedPubMedCentralGoogle Scholar
  58. Woese CR, Kandler O, Wheelis M (1990) Towards a natural system of nomenclature of organisms: proposal for the domains Archaea, Bacteria, and Eucarya. Proc Natl Acad Sci (USA) 87:4576–4579CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Pat M. Fidopiastis
    • 1
  • Daniel J. Bezdek
    • 2
  • Michael H. Horn
    • 2
  • Judith S. Kandel
    • 2
  1. 1.Department of Biological ScienceSkidmore CollegeSaratoga SpringsUSA
  2. 2.Department of Biological ScienceCalifornia State UniversityFullertonUSA

Personalised recommendations