Microbial characterisation of polyhydroxyalkanoates storing populations selected under different operating conditions using a cell-sorting RT-PCR approach

Paulo C. Lemos, Caterina Levantesi, Luisa S. Serafim, Simona Rossetti, Maria A. M. Reis, Valter Tandoi

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

The identity of polyhydroxyalkanoates (PHA) storing bacteria selected under aerobic dynamic feeding conditions, using propionate as carbon source (reactor P), was determined by applying reverse transcriptase-polymerase chain reaction (RT-PCR) on micromanipulated cells and confirmed by fluorescence in situ hybridisation (FISH). Four genera, Amaricoccus, Azoarcus, Thauera and Paraccoccus were detected, the latter only rarely present. All the biomass was involved in PHA storage as shown by Nile Blue staining. By quantitative FISH, their specific amount was determined in this and two other systems using acetate as the carbon substrate (sequencing batch reactor [SBR] A and A1). SBR A and reactor P had the same sludge retention time (SRT, 10 days), while reactor A1 was operated with the SRT of 1 day and the double organic loading rate (OLR). Systems fed with acetate (41.1 ± 2.2 and 49.4 ± 1.4% total Bacteria, for A and A1, respectively) became enriched in Thauera independently on the SRT and OLR, while it was only present in a minor amount when propionate was used as a substrate (1.9 ± 0.2% total Bacteria). Amaricoccus was present in both reactors operated at 10 days SRT, favoured in the one fed with propionate (61.4 ± 1.9% total bacteria), and almost completely removed at the SRT of 1 day. Azoarcus cells were found in all the analysed systems (3.9 ± 0.3, 23.3 ± 1.5 and 45.9 ± 1.5 for P, A and A1, respectively), while Paracoccus was scarcely present.

Original languageEnglish
Pages (from-to)351-360
Number of pages10
JournalApplied Microbiology and Biotechnology
Volume78
Issue number2
DOIs
Publication statusPublished - 1 Feb 2008

Fingerprint

Polyhydroxyalkanoates
Reverse Transcriptase Polymerase Chain Reaction
Thauera
Azoarcus
Propionates
Bacteria
Fluorescence In Situ Hybridization
Population
Acetates
Paracoccus
Carbon
Sewage
Biomass
Staining and Labeling

Keywords

  • Aerobic dynamic feeding
  • FISH
  • Micromanipulation
  • Mixed cultures
  • PHA production
  • RT-PCR

Cite this

@article{fcb6b04d26874240b33e91df5babc8ad,
title = "Microbial characterisation of polyhydroxyalkanoates storing populations selected under different operating conditions using a cell-sorting RT-PCR approach",
abstract = "The identity of polyhydroxyalkanoates (PHA) storing bacteria selected under aerobic dynamic feeding conditions, using propionate as carbon source (reactor P), was determined by applying reverse transcriptase-polymerase chain reaction (RT-PCR) on micromanipulated cells and confirmed by fluorescence in situ hybridisation (FISH). Four genera, Amaricoccus, Azoarcus, Thauera and Paraccoccus were detected, the latter only rarely present. All the biomass was involved in PHA storage as shown by Nile Blue staining. By quantitative FISH, their specific amount was determined in this and two other systems using acetate as the carbon substrate (sequencing batch reactor [SBR] A and A1). SBR A and reactor P had the same sludge retention time (SRT, 10 days), while reactor A1 was operated with the SRT of 1 day and the double organic loading rate (OLR). Systems fed with acetate (41.1 ± 2.2 and 49.4 ± 1.4{\%} total Bacteria, for A and A1, respectively) became enriched in Thauera independently on the SRT and OLR, while it was only present in a minor amount when propionate was used as a substrate (1.9 ± 0.2{\%} total Bacteria). Amaricoccus was present in both reactors operated at 10 days SRT, favoured in the one fed with propionate (61.4 ± 1.9{\%} total bacteria), and almost completely removed at the SRT of 1 day. Azoarcus cells were found in all the analysed systems (3.9 ± 0.3, 23.3 ± 1.5 and 45.9 ± 1.5 for P, A and A1, respectively), while Paracoccus was scarcely present.",
keywords = "Aerobic dynamic feeding, FISH, Micromanipulation, Mixed cultures, PHA production, RT-PCR",
author = "Lemos, {Paulo C.} and Caterina Levantesi and Serafim, {Luisa S.} and Simona Rossetti and Reis, {Maria A. M.} and Valter Tandoi",
note = "Acknowledgements This research was developed within the agreement for Scientific and Technological Cooperation between the Gabinete de Rela{\cc}{\~o}es Internacionais da Ci{\^e}ncia e do Ensino Superior—GRICES (Portugal) and National Research Council— CNR (Italy). The authors acknowledge the financial support of the Funda{\cc}{\~a}o para a Ci{\^e}ncia e Tecnologia (FCT) through the project POCI/BIO/55789/2004 and EU Integrated Project Contract no. 026515. Lu{\'i}sa S. Serafim acknowledges Funda{\cc}{\~a}o para a Ci{\^e}ncia e Tecnologia for grant SFRH/BPD/14663/2003.",
year = "2008",
month = "2",
day = "1",
doi = "10.1007/s00253-007-1301-5",
language = "English",
volume = "78",
pages = "351--360",
journal = "Applied Microbiology and Biotechnology",
issn = "0175-7598",
publisher = "Springer Science Business Media",
number = "2",

}

TY - JOUR

T1 - Microbial characterisation of polyhydroxyalkanoates storing populations selected under different operating conditions using a cell-sorting RT-PCR approach

AU - Lemos, Paulo C.

AU - Levantesi, Caterina

AU - Serafim, Luisa S.

AU - Rossetti, Simona

AU - Reis, Maria A. M.

AU - Tandoi, Valter

N1 - Acknowledgements This research was developed within the agreement for Scientific and Technological Cooperation between the Gabinete de Relações Internacionais da Ciência e do Ensino Superior—GRICES (Portugal) and National Research Council— CNR (Italy). The authors acknowledge the financial support of the Fundação para a Ciência e Tecnologia (FCT) through the project POCI/BIO/55789/2004 and EU Integrated Project Contract no. 026515. Luísa S. Serafim acknowledges Fundação para a Ciência e Tecnologia for grant SFRH/BPD/14663/2003.

PY - 2008/2/1

Y1 - 2008/2/1

N2 - The identity of polyhydroxyalkanoates (PHA) storing bacteria selected under aerobic dynamic feeding conditions, using propionate as carbon source (reactor P), was determined by applying reverse transcriptase-polymerase chain reaction (RT-PCR) on micromanipulated cells and confirmed by fluorescence in situ hybridisation (FISH). Four genera, Amaricoccus, Azoarcus, Thauera and Paraccoccus were detected, the latter only rarely present. All the biomass was involved in PHA storage as shown by Nile Blue staining. By quantitative FISH, their specific amount was determined in this and two other systems using acetate as the carbon substrate (sequencing batch reactor [SBR] A and A1). SBR A and reactor P had the same sludge retention time (SRT, 10 days), while reactor A1 was operated with the SRT of 1 day and the double organic loading rate (OLR). Systems fed with acetate (41.1 ± 2.2 and 49.4 ± 1.4% total Bacteria, for A and A1, respectively) became enriched in Thauera independently on the SRT and OLR, while it was only present in a minor amount when propionate was used as a substrate (1.9 ± 0.2% total Bacteria). Amaricoccus was present in both reactors operated at 10 days SRT, favoured in the one fed with propionate (61.4 ± 1.9% total bacteria), and almost completely removed at the SRT of 1 day. Azoarcus cells were found in all the analysed systems (3.9 ± 0.3, 23.3 ± 1.5 and 45.9 ± 1.5 for P, A and A1, respectively), while Paracoccus was scarcely present.

AB - The identity of polyhydroxyalkanoates (PHA) storing bacteria selected under aerobic dynamic feeding conditions, using propionate as carbon source (reactor P), was determined by applying reverse transcriptase-polymerase chain reaction (RT-PCR) on micromanipulated cells and confirmed by fluorescence in situ hybridisation (FISH). Four genera, Amaricoccus, Azoarcus, Thauera and Paraccoccus were detected, the latter only rarely present. All the biomass was involved in PHA storage as shown by Nile Blue staining. By quantitative FISH, their specific amount was determined in this and two other systems using acetate as the carbon substrate (sequencing batch reactor [SBR] A and A1). SBR A and reactor P had the same sludge retention time (SRT, 10 days), while reactor A1 was operated with the SRT of 1 day and the double organic loading rate (OLR). Systems fed with acetate (41.1 ± 2.2 and 49.4 ± 1.4% total Bacteria, for A and A1, respectively) became enriched in Thauera independently on the SRT and OLR, while it was only present in a minor amount when propionate was used as a substrate (1.9 ± 0.2% total Bacteria). Amaricoccus was present in both reactors operated at 10 days SRT, favoured in the one fed with propionate (61.4 ± 1.9% total bacteria), and almost completely removed at the SRT of 1 day. Azoarcus cells were found in all the analysed systems (3.9 ± 0.3, 23.3 ± 1.5 and 45.9 ± 1.5 for P, A and A1, respectively), while Paracoccus was scarcely present.

KW - Aerobic dynamic feeding

KW - FISH

KW - Micromanipulation

KW - Mixed cultures

KW - PHA production

KW - RT-PCR

UR - http://www.scopus.com/inward/record.url?scp=38649129394&partnerID=8YFLogxK

U2 - 10.1007/s00253-007-1301-5

DO - 10.1007/s00253-007-1301-5

M3 - Article

VL - 78

SP - 351

EP - 360

JO - Applied Microbiology and Biotechnology

JF - Applied Microbiology and Biotechnology

SN - 0175-7598

IS - 2

ER -