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Title: Microbial Diversity in a Dye Treating SBR


1
Microbial Diversityin a Dye Treating SBR
by Dr. Naeem ud din Islamia College Peshawar
2
Biotreatment Different modes
B isolated organisms
A using mixed culture
C isolated enzymes
Dyes are hard to degrade, and often result in
harmful intermediates
3
(No Transcript)
4
  • A Specially Designed Airlift BR from a previous
    Experiment for SND achieving was used
  • The Nitrogen Removing Process was well
    established in that Reactor
  • 93 of Ammonia and COD at an HRT of 12 hrs.

5
Table 1. Physical and Operational Conditions of the SBR Table 1. Physical and Operational Conditions of the SBR
Parameter Value
Working volume (L) Temperature (oC)Dissolved oxygen (mg/l) pH of bioreactor Aeration No aeration(minutes) 3.5 25 - 30 0.05 - 2.0 6.5-8.6 30 120
6
The NITRIFYING MEDIUM
Constituents Quantity
NH4Cl (mg N L-1 ) 120
NaCl (mg L-1 ) 1000
C6H12O6 (mg/L) 1000
FeSO4 (mg L-1 ) 55.00
K2HPO4 (mg L-1 ) 140.00
CaCO3(g L-1 ) 2.00
Trace metal solution(ml/L) 2
Yeast Extract(mg L-1 ) 10
pH 7.8
g/l MgSO47H2O 5, FeCl24H2O 6, COCl2 0.88, H3BO3 0.1, ZnSO47H2O 0.1, CuSO4 0.05, NiSO4 1, MnCl2 5, (NH4)6MO7O244H2O, 0.64 and CaCl22H2O 5. g/l MgSO47H2O 5, FeCl24H2O 6, COCl2 0.88, H3BO3 0.1, ZnSO47H2O 0.1, CuSO4 0.05, NiSO4 1, MnCl2 5, (NH4)6MO7O244H2O, 0.64 and CaCl22H2O 5.
7
MG dye-
  • textile industry, biological stain and
    antifungal.
  • phytotoxic, a respiratory poison, and teratogen

8
  • This SBR was subjected to gradually increasing
    dye concentration
  • Optimization was achieved at a dye concentration
    of 25 mg/l and increased HRT of 36 hrs
  • In this experiment we used the activated sludge
    as a renewable biological resource to adsorb the
    usual environmental concentrations of the MG dye.

9
Synthetic DYE CONTAINING wastewater composition
Constituents Quantity
NH4Cl (mg N L-1 ) 120
NaCl (mg L-1 ) 1000
C6H12O6 (mg/L) 1000
FeSO4 (mg L-1 ) 55.00
K2HPO4 (mg L-1 ) 140.00
CaCO3(g L-1 ) 2.00
Trace metal solution(ml/L) 2
Yeast Extract(mg L-1 ) 10
MG (mg L-1 ) 25
pH 7.8
g/l MgSO47H2O 5, FeCl24H2O 6, COCl2 0.88, H3BO3 0.1, ZnSO47H2O 0.1, CuSO4 0.05, NiSO4 1, MnCl2 5, (NH4)6MO7O244H2O, 0.64 and CaCl22H2O 5. g/l MgSO47H2O 5, FeCl24H2O 6, COCl2 0.88, H3BO3 0.1, ZnSO47H2O 0.1, CuSO4 0.05, NiSO4 1, MnCl2 5, (NH4)6MO7O244H2O, 0.64 and CaCl22H2O 5.
10
  • In that optimized state
  • The Color and COD removal was 80
  • ammonia removal declined to 70 .
  • Biomass, 4 0.7 to 6 0.5 gm/l, SVI was in the
    range of 30 to 65 ml/gm

11
COD Color removal
12
?max 618 nm
UV-Vis spectrophotometric scan of the
biodecolorization of malachite green.
13
Correlation between ammonia, biomass, dye
concentration and OUR
14
  • Knowledge about the microbial community in a dye
    treating reactor would be useful in association
    with operational conditions, to eliminate the
    pollutants efficiently.
  • likely to cause the domination of certain
    groups of bacteria
  • This aspect inspired our interest to know the
    microbial community evolved under the selective
    pressure of the Dye in the SBR.

15
Microbial community structure in the Dye Treating
SBR Sludge
  • 16S rRNA gene Library
  • Phylogenetic Analysis

16
  • PCR-amplification, clone library construction and
    sequencing
  • Bacterial universal primers
  • 27F (3'-AGAGTTTGATCATGGCTCAG-5') and
  • 1492R (3'-TACGGYTACCTTGTTACGACTT-5') were used
    for amplification.

17
BLAST Analysis of the OTUs (culture-
independent)
18
  • Phylogenetic analysis
  • The obtained sequences were edited and
    aligned using the BioEdit software and CLUSTAL_W
    program (Thompson, 199724).
  • The sequences were compared to the known GenBank
    sequences using Basic Local Alignment Search Tool
    (BLAST).
  • Phylogenetic trees were constructed by
    neighbor-joining method with the MEGA package .
    Identical sequences were recognized by
    phylogenetic tree analysis.

19
  • Phylo-genetic analysis
  • If the sequences similarity was more than 97 ,
    they were considered as identical and used for
    further phylogenetic analysis as an operational
    taxonomic unit (OTU).

20
Culture-Independent Phylogenetic tree of the
clones from the dye treating SBR,
21
Phylogenetic distribution profile of microbial
community (Culture Independent) in the SBR.
22
  • Culture-Dependent Method
  • Nineteen isolates were selected from the SBR and
    their 16S rRNA genes were sequenced, and compared
    with similar sequences of the reference organisms
    BLAST search. Figure 6 shows the phylogenetic
    tree based on the culture dependent isolates
    identified with sequences of the NCBI BLAST.
  • Some of the clones identified with the well-known
    biodegraders, the notable being Dokdonella
    koreensis, Rhodobactor, Shingomonas and
    Paracoccus species.

23
Similarity of 16S rRNA gene sequences of the
isolates
Is No
24
Phylogenetic tree of isolates from the dye
treating SBR
The isolates Identified with a- g-
Proteobacteria
25
Phylogenetic distribution, as illustrated by
isolates in the SBR involved in the biotreatment
MG.
All these groups well represented in the
polluted environments
26
  • Table 2 shows the phylogenic affiliation and
    abundance of the clones. The sequences
    identifying with 5 divisions of Proteoabacteria
    i.e ?-, ß-, ?- -proteobacteria and
    Verrucomicrobia groups were obtained. The ß-, and
    ?-proteobacteria were in high abundance, valuing
    24 and 45 of the total clones. The other
    small groups, consisting of ?-,-proteobacteria
    and Verrumicrobia groups, were 4 , 9 , and 2
    respectively. A moderate amount of clones, about
    9 , ranked with the uncultured bacterial strains
    with sequenced data in the NCBI.
  • The similarity of six culture independent
    clones(HT-69, HT-47, HT-51, HT-66, HT-38, HT-7,
    HT-72), to the known sequences in the GenBank was
    lower than 95. Due to difficulty in translating
    16S rRNA gene sequence similarity values into
    nomenclature, it is assumed that similarity
    values to the known sequences below 95 may be
    regarded as evidence of the discovery of novel
    species(3). Thus there is ample possibility of
    unidentified bacteria in the SBR used in the
    present study.

27
inferences
  • SBR with good SVI, effectively removed MG, COD
    and nitrogen up to 25 mg/L dye, above which a
    strong inhibition of these processes was
    observed.
  • The autotrophic nitrifying bacteria were not
    detected at high dye concentration, acting as
    bio-indicators for the MG toxicity. The ammonia
    removal pathway was, however, present, an
    indication of the microbial redundancy.

28
inferences
  • Majority of the sequences identified with the ß-
    and ?-Proteobacteria. pollutant degrading
    bacteria, like rhodobacterales, sphingomonadales
    were in plenty.
  • The first time that MG treated in a nitrifying
    BR, with its inhibitory effects, and microbial
    community monitored.
  • Both culture-dependent and Culture independent
    methods must be used to have a true picuture of
    microbial diversity.

29
References Aksu, Z.(2005). Application of
biosorption for the removal of organic
pollutants a review. Process. biochem. 40,
9971026. Altschul, S. F. Gish, M. W. Myers,
W. Lipman, D.J.(1990). Basic local alignment
search tool. J. mol. biol. 215, 403-410. Amann,
R.I. Ludwig W. Schleifer K.H.
(1995).Phylogenetic identification and in situ
detection of individual microbial cells without
cultivation. FEMS microbiol. rev. 59,
143-169. Azmi, W. Kumar, R. Sani Banerjee,
U.C. (1998). Biodegradation of triphenylmethane
dyes. Enzyme. microb. technol. 22,
185-191. Cabral, G. I. Penha, S. Matos, M.
Santos, A. R Franco, F. Pinheiro, H.M.
(2005). Evaluation of an integrated anaerobic
/aerobic SBR system for the treatment of wool
dyeing effluents. Biodegradation. 16, 81-89 Cha,
C.J. Daniel R.D. Carl, E.C. (2001).
Biotransformation of malachite green by the
fungus Cunninghamella elegans. Appl. Environ.
Microbiol. 67, 4358-4360 Chen, K.C. Wu, J.Y.
Liou D.J. Hwang, S.C.J. (2003). Decolorization
of the textile dyes by newly isolated bacterial
strains. J. biotechnol. 101, 5768. Daneshvar,
N. Ayazloo, M. Khataee. A.R. Pourhassan, M.
(2007) Biological decolorization of dye solution
containing Malachite Green by microalgae
Cosmarium sp. Bioresource Technology. 98,
1176-1182. Govoreanu, R. Seghers, D. Nopens,
I. Clercq, B. D., Saveyn, H. Capalozza, C.
(2003). Linking floc structure and settling
properties to activated sludge populations
dynamics in an SBR..Wat. Sci. Tech. 47, 9-18.
Hitz H.R. Huber, W. Rud, R.H.(1978). The
adsorption of dyes by activated sludge, J. Soc.
Dyers. Colour. 94 7176. Hong, J. Otaki, M.
(2003). Effects of photocatalyis on biological
decolorization reactor and biological activity of
isolated photosynthetic bacteria. J. Biosci.
Bioeng. 96, 298-303. Jiang, H.L. Tay, J.H.
Tay, S.T.L.(2004). Changes in structure, activity
and metabolism of aerobic granules as a microbial
response to high phenol loading. Appl.
Microbiol.Biotechnol. 63, 602608. Kandelbauer,
A. Guebitz G. M.(2005). Bioremediation for the
Decolorization of Textile Dyes A Review.
Environmental Chemistry, Springer Heidelberg
30
  • Kipopoulou, A.M. Zouloulis, A. Samara, C.
    Kouimtzis, Th. (2004). The fate of lindane in the
    conventional activated sludge treatment process,
    Chemosphere. 55, 8191.
  • Kumar, S. Tamura, K. and Nei, M. (2004). MEGA3
    Integrated software for Molecular Evolutionary
    Genetics Analysis and sequence alignment.
    Briefings in Bioinformatics. 5, 150-163.
  • Shi-Quan, N. Jun, F. Ying, J. Yuichi, I.
    Tohru, U. Satoshi, M. (2006). Analysis of
    bacterial community structure in the natural
    Circulation system wastewater bioreactor by using
    a 16S rRNA gene clone liberary. Microbiol.
    Immunol, 50, 937-950.
  • Ong, S-A. Toorisaka E. Hirata, M. Hano, T.
    (2005). Treatment of azo dye Orange II in aerobic
    and anaerobic-SBR systems. Process Biochem. 40,
    2907-2914
  • Rai, H.S. Singh S. Cheema P.P.S. Bansal T.K.
    Banerjee U.C. (2007). Decolorization of
    triphenylmethane dye-bath effluent in an
    integrated two-stage anaerobic reactor. J.
    Environ. Manage. 83, 290297
  • Saitou, N. Nei, M. (1987). The neighbour-joining
    method a new method for reconstructing
    phylogenetic trees. Mol. Biol. Evol. 4, 406-425.
  • Sani, R.K. Banerjee, U. C. (1999).
    Decolorization of triphenylmethane dyes and
    textile dye-stuff effluent by Kurthia sp. .
    Enzyme. microb. technol. 24, 433-437.
  • American Public Health Association.(1998).
    Standard Methods for the Examination of Water and
    Wastewater. 19th ed. Washington, DC, USA.
  • Thompson, J.D. Gibson, T.J. Plewniak,F.
    Jeanmougin, F. Higgins, D.G. (1997). The
    CLUSTAL_X windows interface Flexible strategies
    for multiple sequence alignment aided by quality
    analysis tools. Nucleic Acids Res. 25, 4876-82.
  • Xu, P. Qian X. M. Wang, Y. X. Xu. Y.B. (2004).
    Modeling for waste water treatment by
    Rhodopseudomonas palustris Y6 immobilized on
    fiber in a columnar bioreactor. Appl Microbiol
    Biotechnol . 44, 676-682.
  • Zilouei, H. Soares, A. Murto, M. (2006).
    Influence of temperature on process efficiency
    and microbial community response during the
    biological removal of chlorophenols in a
    packed-bed bioreactor. Appl Microbiol Biotechnol.
    72, 591-599.

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