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Aquaculture
Volume 215, Issues 1-4, 10 January 2003, Pages 187-202

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DOI: 10.1016/S0044-8486(02)00372-1
PII: S0044-8486(02)00372-1
Copyright © 2003 Elsevier Science B.V. All rights reserved.

Characterization of the microbial community and nitrogen transformation processes associated with moving bed bioreactors in a closed recirculated mariculture system

Yossi Tal^a, Joy E. M. Watts^a, Susan B. Schreier^a, Kevin R. Sowers^a and Harold J. Schreier^a^,^b^,^,

^a Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 701 E. Pratt St., Baltimore, MD 21202, USA
^b Department of Biological Sciences, University of Maryland Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250, USA

Received 25 February 2002; revised 3 July 2002; accepted 19 July 2002. Available online 6 November 2002.

Abstract

The microbial consortium of a moving bed bioreactor (MBB) connected to a marine recirculating aquaculture system was examined by denaturing gradient gel electrophoresis (DGGE) of amplified 16S rRNA gene fragments. Both ammonia and nitrite oxidizers, Nitrosomonas cryotolerans and Nitrospira marina, respectively, were found associated with the marine system as well as a number of heterotrophic bacteria, including Pseudomonas sp. and Sphingomonas sp. In addition, two Planctomycetes sp. were detected in the system suggesting the capability for anaerobic ammonia oxidation (anammox). The potential for carrying out different nitrogen transformation processes––nitrification, denitrification and anammox––by the bead consortium in both low and high organic load MBBs was measured by short-term batch incubation. Beads with a high organic load exhibited a lower nitrification rate (25 mg NH₃–N/m²/h) than low organic load beads (31.5 mg NH₃–N/m²/h) as well as the ability to carry out denitrification and anammox processes. The potential of using MBBs to induce different nitrogen transformation processes was evaluated, and it was found that this type of bioreactor has the capability to serve as a platform for mediating desired anoxic processes such as denitrification and anammox.

Author Keywords: Nitrification; Anammox; Denitrification; Microbial community profile

Article Outline

1. Introduction
2. Materials and methods 2.1. Aquaculture facilities and filter setup 2.2. Batch experiments 2.3. Chemical and physical analyses 2.4. Bead dry weight 2.5. DNA extraction and polymerase chain reaction (PCR) amplification 2.6. DGGE analysis 2.7. Cloning, sequencing, and database analysis
3. Results 3.1. MBB bead biofilm formation and characteristics 3.2. Bacterial diversity 3.3. Inorganic nitrogen transformation processes 3.3.1. Nitrification potential 3.3.2. Denitrification potential 3.3.3. Anaerobic ammonia oxidation potential
4. Discussion
5. Conclusion
Acknowledgements
References

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Fig. 1. Dry organic weight of beads. Dry weights for beads in high (

) and low (

) organic load filters during a time-course of incubation was determined as described in Materials and methods.

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Fig. 2. Biofilm formation on beads from different organic load MBBs. Photograph of a representative bead from the low (B) and high (C) organic load systems are shown as well as an untreated bead (A). Magnification (×3) of a small section of the matrix (boxed area) is shown under each bead. The dark area between bead segments is due to biofilm formation.

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Fig. 3. Oxygen consumption pattern of filters. Oxygen levels for high (

) and low (

) organic load systems were measured as described in Materials and methods.

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Fig. 4. DGGE analysis of PCR-amplified DNA extracted from filter biofilms. Samples from low (A) and high (B) organic load filters were compared to a freshwater filter sample (C). Also shown (D) is a collection of individual fragments that were isolated from these samples, cloned and characterized. Numbers and arrows correlate with the identification listed in Table 1.

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Fig. 5. Patterns for inorganic nitrogen consumption and accumulation during nitrification conditions. NH₃–N ([&z.cirf;]), NO₂–N (

) and NO₃–N (

) levels were measured as described in Materials and methods.

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Fig. 6. Nitrate and nitrite concentration during incubation of MBBs under denitrification conditions. Nitrate (

) and nitrite (

) concentrations were determined as described in Materials and methods and incubations were done: (A) with high load beads in the absence of an external carbon source; (B) during a second incubation of high load beads without the addition of external carbon source; (C) with low load beads in the presence of 10 mM acetate as carbon source; and (D) with high load beads in the presence of 10 mM of acetate as carbon source.

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Fig. 7. Anammox potential for the high organic load filter. Ammonia (A) and nitrite (B) removal rates of high load beads were measured during incubations under anammox conditions as described in Materials and methods. For A, incubations were done in the presence of ammonia and nitrite (open boxes) or with ammonia alone (closed boxes); for B, incubations were done in the presence of ammonia and nitrite (open boxes) or with nitrite alone (closed boxes).

Table 1. Characteristics of DNA fragments from DGGE gels

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Table 2. Ammonia removals rates of different filter configurations

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Corresponding author. Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 701 E. Pratt St., , Baltimore, MD 21202, , USA. Tel.: +1-410-234-8874; fax: +1-410-234-8896; email: schreier@umbi.umd.edu

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Aquaculture
Volume 215, Issues 1-4, 10 January 2003, Pages 187-202

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