Organic loading rates affect composition of soil-derived bacterial communities during continuous, fermentative biohydrogen production

Yonghua Luo, Husen Zhang, Michael Salerno, Bruce Ernest Logan, Maryann Victoria Bruns

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Bacterial community composition during steady-state, fermentative H2 production was compared across a range of organic loading rates (OLRs) of 0.5-19 g COD l-1 h-1 in a 2-l continuous flow reactor at 30 °C. The varied OLRs were achieved with glucose concentrations of 2.5-10 g l-1 and hydraulic retention times of 1-10 h. The synthetic wastewater feed was amended with l-cysteine and maintained at a pH of 5.5. For each run at a given glucose concentration, the reactor was inoculated with an aliquot of well-mixed agricultural topsoil that had been heat-treated to reduce numbers of vegetative cells. At OLRs less than 2 g COD l-1 h-1, DNA sequences from ribosomal RNA intergenic spacer analysis profiles revealed more diverse and variable populations (Selenomonas, Enterobacter, and Clostridium spp.) than were observed above 2 g COD l-1 h-1 (Clostridium spp. only). An isolate, LYH1, was cultured from a reactor sample (10 g glucose l-1 at a 10-h HRT) on medium containing l-cysteine. In confirming H2 production by LYH1 in liquid batch culture, lag periods for H2 production in the presence and absence of l-cysteine were 5 and 50 h, respectively. The 16S rRNA gene sequence of LYH1 indicated that the isolate was a Clostridium sp. affiliated with RNA subcluster Ic, with >99% similarity to Clostridium sp. FRB1. In fluorescent in situ hybridization tests, an oligonucleotide probe complementary to the 16S rRNA of LYH1 hybridized with 90% of cells observed at an OLR of 2 g COD h-1, compared to 26% of cells at an OLR of 0.5 g COD l-1 h-1. An OLR of 2 g COD l-1 h-1 appeared to be a critical threshold above which clostridia were better able to outcompete Enterobacteriaceae and other organisms in the mixed soil inoculum. Our results are discussed in light of other biohydrogen studies employing pure cultures and mixed inocula.

Original languageEnglish (US)
Pages (from-to)6566-6576
Number of pages11
JournalInternational Journal of Hydrogen Energy
Volume33
Issue number22
DOIs
StatePublished - Jan 1 2008

Fingerprint

Clostridium
loading rate
crack opening displacement
soils
Soils
cysteine
Glucose
Chemical analysis
inoculum
glucose
reactors
cells
ribonucleic acids
Oligonucleotides
DNA sequences
RNA
oligonucleotides
organisms
Wastewater
hydraulics

All Science Journal Classification (ASJC) codes

  • Renewable Energy, Sustainability and the Environment
  • Fuel Technology
  • Condensed Matter Physics
  • Energy Engineering and Power Technology

Cite this

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title = "Organic loading rates affect composition of soil-derived bacterial communities during continuous, fermentative biohydrogen production",
abstract = "Bacterial community composition during steady-state, fermentative H2 production was compared across a range of organic loading rates (OLRs) of 0.5-19 g COD l-1 h-1 in a 2-l continuous flow reactor at 30 °C. The varied OLRs were achieved with glucose concentrations of 2.5-10 g l-1 and hydraulic retention times of 1-10 h. The synthetic wastewater feed was amended with l-cysteine and maintained at a pH of 5.5. For each run at a given glucose concentration, the reactor was inoculated with an aliquot of well-mixed agricultural topsoil that had been heat-treated to reduce numbers of vegetative cells. At OLRs less than 2 g COD l-1 h-1, DNA sequences from ribosomal RNA intergenic spacer analysis profiles revealed more diverse and variable populations (Selenomonas, Enterobacter, and Clostridium spp.) than were observed above 2 g COD l-1 h-1 (Clostridium spp. only). An isolate, LYH1, was cultured from a reactor sample (10 g glucose l-1 at a 10-h HRT) on medium containing l-cysteine. In confirming H2 production by LYH1 in liquid batch culture, lag periods for H2 production in the presence and absence of l-cysteine were 5 and 50 h, respectively. The 16S rRNA gene sequence of LYH1 indicated that the isolate was a Clostridium sp. affiliated with RNA subcluster Ic, with >99{\%} similarity to Clostridium sp. FRB1. In fluorescent in situ hybridization tests, an oligonucleotide probe complementary to the 16S rRNA of LYH1 hybridized with 90{\%} of cells observed at an OLR of 2 g COD h-1, compared to 26{\%} of cells at an OLR of 0.5 g COD l-1 h-1. An OLR of 2 g COD l-1 h-1 appeared to be a critical threshold above which clostridia were better able to outcompete Enterobacteriaceae and other organisms in the mixed soil inoculum. Our results are discussed in light of other biohydrogen studies employing pure cultures and mixed inocula.",
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Organic loading rates affect composition of soil-derived bacterial communities during continuous, fermentative biohydrogen production. / Luo, Yonghua; Zhang, Husen; Salerno, Michael; Logan, Bruce Ernest; Bruns, Maryann Victoria.

In: International Journal of Hydrogen Energy, Vol. 33, No. 22, 01.01.2008, p. 6566-6576.

Research output: Contribution to journalArticle

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T1 - Organic loading rates affect composition of soil-derived bacterial communities during continuous, fermentative biohydrogen production

AU - Luo, Yonghua

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N2 - Bacterial community composition during steady-state, fermentative H2 production was compared across a range of organic loading rates (OLRs) of 0.5-19 g COD l-1 h-1 in a 2-l continuous flow reactor at 30 °C. The varied OLRs were achieved with glucose concentrations of 2.5-10 g l-1 and hydraulic retention times of 1-10 h. The synthetic wastewater feed was amended with l-cysteine and maintained at a pH of 5.5. For each run at a given glucose concentration, the reactor was inoculated with an aliquot of well-mixed agricultural topsoil that had been heat-treated to reduce numbers of vegetative cells. At OLRs less than 2 g COD l-1 h-1, DNA sequences from ribosomal RNA intergenic spacer analysis profiles revealed more diverse and variable populations (Selenomonas, Enterobacter, and Clostridium spp.) than were observed above 2 g COD l-1 h-1 (Clostridium spp. only). An isolate, LYH1, was cultured from a reactor sample (10 g glucose l-1 at a 10-h HRT) on medium containing l-cysteine. In confirming H2 production by LYH1 in liquid batch culture, lag periods for H2 production in the presence and absence of l-cysteine were 5 and 50 h, respectively. The 16S rRNA gene sequence of LYH1 indicated that the isolate was a Clostridium sp. affiliated with RNA subcluster Ic, with >99% similarity to Clostridium sp. FRB1. In fluorescent in situ hybridization tests, an oligonucleotide probe complementary to the 16S rRNA of LYH1 hybridized with 90% of cells observed at an OLR of 2 g COD h-1, compared to 26% of cells at an OLR of 0.5 g COD l-1 h-1. An OLR of 2 g COD l-1 h-1 appeared to be a critical threshold above which clostridia were better able to outcompete Enterobacteriaceae and other organisms in the mixed soil inoculum. Our results are discussed in light of other biohydrogen studies employing pure cultures and mixed inocula.

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