Microbial Ecology

, Volume 51, Issue 3, pp 277–284 | Cite as

Genetic and Physiological Characterization of the Intestinal Bacterial Microbiota of Bluegill (Lepomis macrochirus) with Three Different Feeding Habits

  • Kimiko UchiiEmail author
  • Kazuaki Matsui
  • Ryuji Yonekura
  • Katsuji Tani
  • Takehiko Kenzaka
  • Masao Nasu
  • Zen'ichiro Kawabata


Bluegill (Lepomis macrochirus) in Lake Biwa, Japan, feed on benthic invertebrates (benthivorous type), aquatic plants (herbivorous type), and zooplankton (planktivorous type). To evaluate the effect of food on intestinal bacterial microbiota, we characterized and compared the intestinal microbiota of these three types of bluegill in terms of community-level physiological profile (CLPP) and genetic structure. The CLPP was analyzed using Biolog MicroPlates (Biolog, Inc., Hayward, CA, USA), and multivariate analysis of variance revealed that the CLPP of intestinal microbiota differed significantly between any pairs of the three types of bluegill. The genetic profiles were analyzed by temperature gradient gel electrophoresis of polymerase chain reaction (PCR)-amplified 16S rDNA fragments, and multidimensional scaling indicated the existence of specific intestinal bacterial structures for both the benthivorous and the planktivorous types. These results suggest that the host's feeding habit can be one factor controlling the intestinal microbiota of fish in the natural environment.


Bacterial Community Intestinal Microbiota Benthic Invertebrate Intestinal Bacterium Terrestrial Insect 
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.



We greatly acknowledge S. Gritters and K. Aulwes, Iowa Department of Natural Resources, for fish sampling in Iowa. We thank N. Yamaguchi, C. Sakamoto, F. Maruyama, and other members of Osaka University for their help in the TGGE experiments. We also thank M. Yuma for reviewing an earlier draft of this manuscript, and M. Honjo and M. Ueki for helpful comments and technical assistance. This research was partly supported by the Grant for the Biodiversity Research of the 21st Century COE (A14). The study was also partly supported by the JSPS Grant-in Aid for Basic Research (A) (16207001).


  1. 1.
    Bochner, BR (1989) Breathprints at the microbial level. ASM News 55: 536–539Google Scholar
  2. 2.
    Cahill, MM (1990) Bacterial flora of fishes: A review. Microb Ecol 19: 21–41CrossRefGoogle Scholar
  3. 3.
    Campbell, AC, Buswell, JA (1983) The intestinal microflora of farmed Dover sole (Solea solea) at different stages of fish development. J Appl Bacteriol 55: 215–223Google Scholar
  4. 4.
    Don, RH, Cox, PT, Wainwright, BJ, Baker, K, Mattick, JS (1991) Touchdown PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res 19: 4008PubMedCrossRefGoogle Scholar
  5. 5.
    Ferrari, VC, Hollibaugh, JT (1999) Distribution of microbial assemblages in the Central Arctic Ocean Basin studied by PCR/DGGE: Analysis of a large data set. Hydrobiologia 401: 55–68CrossRefGoogle Scholar
  6. 6.
    Garland, JL, Mills, AL (1991) Classification and characterization of heterotrophic microbial communities on the basis of patterns of community-level sole-carbon-source utilization. Appl Environ Microbiol 57: 2351–2359PubMedGoogle Scholar
  7. 7.
    Hansen, GH, Olafsen, JA (1999) Bacterial interactions in early life stages of marine cold water fish. Microb Ecol 38: 1–26CrossRefPubMedGoogle Scholar
  8. 8.
    Harmsen, HJM, Wildeboer-Veloo, ACM, Raangs, GC, Wagendorp, AA, Klijn, N, Bindels, JG, Welling, GW (2000) Analysis of intestinal flora development in breast-fed and formula-fed infants by using molecular identification and detection methods. J Pediatr Gastroenterol Nutr 30: 61–67CrossRefPubMedGoogle Scholar
  9. 9.
    Hentges, DJ (1980) Does diet influence human fecal microflora composition. Nutr Rev 38: 329–336PubMedCrossRefGoogle Scholar
  10. 10.
    Hill, MJ (1981) Diet and the human intestinal bacterial flora. Cancer Res 41: 3778–3780PubMedGoogle Scholar
  11. 11.
    Holben, WE, Williams, P, Saarinen, M, Sarkilahti, LK, Apajalahti, JHA (2002) Phylogenetic analysis of intestinal microflora indicates a novel Mycoplasma phylotype in farmed and wild salmon. Microb Ecol 44: 175 –185CrossRefPubMedGoogle Scholar
  12. 12.
    Hynes, HBN (1950) The food of fresh-water sticklebacks (Gasterosteus aculeatus and Pygosteus pugitius), with a review of methods used in studies of the food of fishes. J Anim Ecol 19: 36–58CrossRefGoogle Scholar
  13. 13.
    Iwamoto, T, Tani, K, Nakamura, K, Suzuki, Y, Kitagawa, M, Eguchi, M, Nasu, M (2000) Monitoring impact of in situ biostimulation treatment on groundwater bacterial community by DGGE. FEMS Microbiol Ecol 32: 129–141PubMedCrossRefGoogle Scholar
  14. 14.
    Konopka, A, Oliver, L, Turco Jr, RF (1998) The use of carbon substrate utilization patterns in environmental and ecological microbiology. Microb Ecol 35: 103–115CrossRefPubMedGoogle Scholar
  15. 15.
    Mackie, RI, Sghir, A, Gaskins, HR (1999) Developmental microbial ecology of the neonatal gastrointestinal tract. Am J Clin Nutr 69: 1035S–1045SPubMedGoogle Scholar
  16. 16.
    Muyzer, G, de Waal, EC, Uitterlinden, AG (1993) Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol 59: 695–700PubMedGoogle Scholar
  17. 17.
    Muyzer, G, Smalla, K (1998) Application of denaturing gradient gel electrophoresis (DGGE) and temperature gradient gel electrophoresis (TGGE) in microbial ecology. Antonie Van Leeuwenhoek 73: 127–141CrossRefPubMedGoogle Scholar
  18. 18.
    Myers, RM, Fischer, SG, Lerman, LS, Maniatis, T (1985) Nearly all single base substitutions in DNA fragments joined to a GC-clamp can be detected by denaturing gradient gel electrophoresis. Nucleic Acids Res 13: 3131–3145PubMedCrossRefGoogle Scholar
  19. 19.
    Nedoluha, PC, Westhoff, D (1997) Microbiology of striped bass grown in three aquaculture systems. Food Microbiol 14: 255–264CrossRefGoogle Scholar
  20. 20.
    Schmidt-Nielsen, K (1990) Animal Physiology: Adaptation and Environment, 4th ed. Cambridge University Press, New York, p 319Google Scholar
  21. 21.
    Simpson, JM, McCracken, VJ, White, BA, Gaskins, HR, Mackie, RI (1999) Application of denaturant gradient gel electrophoresis for the analysis of the porcine gastrointestinal microbiota. J Microbiol Methods 36: 167–179CrossRefPubMedGoogle Scholar
  22. 22.
    Sugita, H, Kawasaki, J, Deguchi, Y (1997) Production of amylase by the intestinal microflora in cultured freshwater fish. Lett Appl Microbiol 24: 105–108CrossRefPubMedGoogle Scholar
  23. 23.
    Sugita, H, Miyajima, C, Deguchi, Y (1991) The vitamin B12-producing ability of the intestinal microflora of freshwater fish. Aquaculture 92: 267–276CrossRefGoogle Scholar
  24. 24.
    Swynnerton, GH, Worthington, EB (1940) Note on the food of fish in Haweswater (Westmorland). J Anim Ecol 9: 183–187CrossRefGoogle Scholar
  25. 25.
    Tsai, YL, Olson, BH (1991) Rapid method for direct extraction of DNA from soil and sediments. Appl Environ Microbiol 57: 1070–1074PubMedGoogle Scholar
  26. 26.
    Yonekura, R, Nakai, K, Yuma, M (2002) Trophic polymorphism in introduced bluegill in Japan. Ecol Res 17: 49–57CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Kimiko Uchii
    • 1
    Email author
  • Kazuaki Matsui
    • 2
  • Ryuji Yonekura
    • 3
  • Katsuji Tani
    • 4
  • Takehiko Kenzaka
    • 4
  • Masao Nasu
    • 4
  • Zen'ichiro Kawabata
    • 1
    • 5
  1. 1.Center for Ecological ResearchKyoto UniversityShigaJapan
  2. 2.Laboratory of Environmental Biotechnology, Faculty of EngineeringTohoku Gakuin UniversityTagajoJapan
  3. 3.Gifu Prefectural Research Institute for Freshwater Fish and Aquatic EnvironmentsKakamigaharaJapan
  4. 4.Environmental Science and Microbiology, Graduate School of Pharmaceutical SciencesOsaka UniversitySuitaJapan
  5. 5.Research Institute for Humanity and NatureKamigyo-kuJapan

Personalised recommendations