Cited by 37Mine drainage and remediation techniquesMicrobial Community Ecology and PhysiologyMicrobial Fuel Cells and Bioremediation

The DOI matches its Crossref record; 61 of the 62 references it cites were found in a registry or an index; 3 of the 61 sources its claims cite were checked for support.

Microbial Diversity in Sulfate-Reducing Marine Sediment Enrichment Cultures Associated with Anaerobic Biotransformation of Coastal Stockpiled Phosphogypsum (Sfax, Tunisia)

Open access
Zouch, Hana, Karray, Fatma, Armougom, Fabrice, Chifflet, Sandrine, Hirschler-Réa, Agnès, Kharrat, Hanen, Kamoun, Lotfi, Ben Hania, Wajdi, Ollivier, Bernard, Sayadi, Sami, Quéméneur, Marianne·2017·Frontiers in Microbiology

Abstract

Anaerobic biotechnology using sulfate-reducing bacteria (SRB) is a promising alternative for reducing long-term stockpiling of phosphogypsum (PG), an acidic (pH ~3) by-product of the phosphate fertilizer industries containing high amounts of sulfate. The main objective of this study was to evaluate, for the first time, the diversity and ability of anaerobic marine microorganisms to convert sulfate from PG into sulfide, in order to look for marine SRB of biotechnological interest. A series of sulfate-reducing enrichment cultures were performed using different electron donors (i.e. acetate, formate or lactate) and sulfate sources (i.e. sodium sulfate or PG) as electron acceptors. Significant sulfide production was observed from enrichment cultures inoculated with marine sediments, collected near the effluent discharge point of a Tunisian fertilizer industry (Sfax, Tunisia). Sulfate sources impacted sulfide production rates from marine sediments as well as the diversity of SRB species belonging to Deltaproteobacteria. When PG was used as sulfate source, Desulfovibrio species dominated microbial communities of marine sediments, while Desulfobacter species were mainly detected using sodium sulfate. Sulfide production was also affected depending on the electron donor used, with the highest production obtained using formate. In contrast, low sulfide production (acetate-containing cultures) was associated with an increase in the population of Firmicutes. These results suggested that marine Desulfovibrio species, to be further isolated, are potential candidates for bioremediation of PG by immobilizing metals and metalloids thanks to sulfide production by these SRB.

Overview

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Written from the paper’s full text.

TL;DR

This study evaluated sulfate-reducing enrichment cultures from Tunisian coastal marine sediments for anaerobic biotransformation of phosphogypsum (PG), showing that sulfide was produced from PG and sodium sulfate. Formate gave the highest H2S production, and the sulfate source shifted SRB communities: Desulfovibrio dominated PG cultures while Desulfobacter were mainly detected with sodium sulfate.

Key findings

  • Sulfide production occurred from marine sediment enrichments with either Na2SO4 or PG; formate gave the highest rates (2.4 ± 0.2 mM/day with Na2SO4 and 3.0 ± 0.2 mM/day with PG), while acetate gave <1 mM/day. In PG cultures, maximum H2S was highest with formate (19.7 ± 3.7 mM), about twice that with acetate or lactate.
  • Sulfate source affected SRB diversity: PG cultures were dominated by Desulfovibrio spp., whereas Desulfobacter spp. were only retrieved in Na2SO4 cultures, suggesting inhibition by PG. Deltaproteobacteria proportions correlated positively with final pH (r = 0.94), maximum H2S (r = 0.83), and maximum H2S rate (r = 0.89); Simpson index correlated negatively with these H2S performance measures.
  • Initial marine sediment had higher microbial diversity than PG: 1,686 OTUs, Shannon 8.98, Simpson 0.99 in sediment versus 224 OTUs, Shannon 4.51, Simpson 0.88 in PG. PG was acidic (pH 3.1), contained 55% sulfate of dry weight, and had low organic carbon (0.53 ± 0.15%).
  • Low sulfide production in acetate + PG cultures was associated with increased Firmicutes: Deltaproteobacteria represented only 10.9%, while Firmicutes were 25.8 ± 3.6%, Fusobacteria 21.6 ± 5.8%, Epsilonproteobacteria 21.1 ± 2.2%, and Spirochaetes 6.6 ± 1.2%.

Methods

  • Samples were collected in March 2016 from the Sfax phosphate fertilizer complex, Tunisia: marine sediment near the acid effluent discharge and PG from a stockpile. Chemical analyses included pH, dissolved oxygen, sulfate, C/H/N/S, total organic carbon, and trace metals by HR-ICP-MS.
  • Duplicate sulfate-reducing enrichment cultures were prepared in 120 mL bottles with basal medium, Na2SO4 (1.58 g/L) or PG (2 g/L) as sulfate source, and acetate (20 mM), lactate (40 mM), or formate (80 mM) as electron donors. Cultures were inoculated with 5 mL marine sediment or non-autoclaved PG, incubated at 37°C for 2 weeks, and monitored for H2S; electron donors and sulfate were analyzed by HPLC and ion chromatography.
  • DNA was extracted from environmental samples and enrichment pellets, and 16S rRNA gene amplicons were sequenced on Illumina MiSeq. QIIME/UCLUST clustered OTUs at 97% identity; diversity indices, phylogenetic trees, ANOVA with Bonferroni post-hoc tests, Spearman correlations, and PCA were used.

Limitations

  • Stated: further work is needed to isolate SRB from marine sediments, especially Desulfovibrionales species, to test their metal resistance and performance in PG biotransformation. The paper also notes that the DNA extraction method following sampling and storage may affect the revealed community structure.
  • Evident: the study measured H2S production and microbial community composition but did not directly measure immobilization or fate of trace metals or radionuclides in the enrichment cultures, despite proposing metal immobilization as a bioremediation mechanism.
  • Evident: results come from short-term (14-day) laboratory microcosms with two biological replicates, so field-scale performance and long-term stability are not established.

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Integrity signals

Registry

Crossref · 10.3389/fmicb.2017.01583

Cites

61 of 61 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Figures

Figure 1 Compositions of microbial communities in the coastal marine sediment and phosphogypsum samples of Sfax (Tunisia). Relative phylogenetic abundance was based on frequencies of 16S rRNA gene sequences affiliated with Archaea and major bacterial phyla or proteobacterial classes in the microbial communities of marine sediment (MS) and phosphogypsum (PG).
Figure 1 Compositions of microbial communities in the coastal marine sediment and phosphogypsum samples of Sfax (Tunisia). Relative phylogenetic abundance was based on frequencies of 16S rRNA gene sequences affiliated with Archaea and major bacterial phyla or proteobacterial classes in the microbial communities of marine sediment (MS) and phosphogypsum (PG).CC BY · PubMed Central
Figure 2 Profiles of hydrogen sulfide production during 14 days of enrichment cultures from coastal marine sediment (as inoculum) using different electron donors (AC, acetate; FOR, formate; LAC, lactate; C, control without electron donor) and sulfate sources: sodium sulfate (A) or phosphogypsum (B). Values are means of two biological replicates ± confidence intervals (error bars).
Figure 2 Profiles of hydrogen sulfide production during 14 days of enrichment cultures from coastal marine sediment (as inoculum) using different electron donors (AC, acetate; FOR, formate; LAC, lactate; C, control without electron donor) and sulfate sources: sodium sulfate (A) or phosphogypsum (B). Values are means of two biological replicates ± confidence intervals (error bars).CC BY · PubMed Central
Figure 3 Compositions of microbial communities in sulfate-reducing enrichment cultures from marine sediment using different electron donors and sulfate sources after 14 days. Relative phylogenetic abundance was based on frequencies of 16S rRNA gene sequences affiliated with Archaea and major bacterial phyla or proteobacterial classes in the microbial communities. The names of enrichment cultures (duplicates 1 and 2) have been abbreviated as follows: SF, SL, SA, and SC for Na2SO4 enrichment cultures with formate, lactate, acetate and without electron donor, respectively; SPF, SPL, SPA, and SPC for PG enrichment cultures with formate, lactate, acetate, and without electron donor, respectively.
Figure 3 Compositions of microbial communities in sulfate-reducing enrichment cultures from marine sediment using different electron donors and sulfate sources after 14 days. Relative phylogenetic abundance was based on frequencies of 16S rRNA gene sequences affiliated with Archaea and major bacterial phyla or proteobacterial classes in the microbial communities. The names of enrichment cultures (duplicates 1 and 2) have been abbreviated as follows: SF, SL, SA, and SC for Na2SO4 enrichment cultures with formate, lactate, acetate and without electron donor, respectively; SPF, SPL, SPA, and SPC for PG enrichment cultures with formate, lactate, acetate, and without electron donor, respectively.CC BY · PubMed Central
Figure 4 Principal Component Analysis (PCA) biplot showing the variation among the enrichment cultures based on hydrogen sulfide production performances and the relative abundance of microbial taxa. Black circles represent PG enrichment cultures and black triangles represent Na2SO4 enrichment cultures. The names of enrichment cultures (duplicates 1 and 2) have been abbreviated as follows: SF, SL, and SA, for Na2SO4 enrichment cultures with formate, lactate, and acetate as electron donors, respectively; SPF, SPL, and SPA, for PG enrichment cultures with formate, lactate, and acetate as electron donors, respectively. Arrows indicate the direction of maximum increase and strength (through the length) of each variable to the overall distribution. The blue arrows are indicators of hydrogen sulfide production (pH, Pmax, Vmax) and the red arrows represent the microbial taxa. Among these latter, α-Prot, β-Prot, δ-Prot, ε-Prot stand for Alphaproteobacteria, Betaproteobacteria, Deltaproteobacteria, and Epsilonproteobacteria. The first two principal axes explained 61.3% of the variance.
Figure 4 Principal Component Analysis (PCA) biplot showing the variation among the enrichment cultures based on hydrogen sulfide production performances and the relative abundance of microbial taxa. Black circles represent PG enrichment cultures and black triangles represent Na2SO4 enrichment cultures. The names of enrichment cultures (duplicates 1 and 2) have been abbreviated as follows: SF, SL, and SA, for Na2SO4 enrichment cultures with formate, lactate, and acetate as electron donors, respectively; SPF, SPL, and SPA, for PG enrichment cultures with formate, lactate, and acetate as electron donors, respectively. Arrows indicate the direction of maximum increase and strength (through the length) of each variable to the overall distribution. The blue arrows are indicators of hydrogen sulfide production (pH, Pmax, Vmax) and the red arrows represent the microbial taxa. Among these latter, α-Prot, β-Prot, δ-Prot, ε-Prot stand for Alphaproteobacteria, Betaproteobacteria, Deltaproteobacteria, and Epsilonproteobacteria. The first two principal axes explained 61.3% of the variance.CC BY · PubMed Central
Figure 5 Maximum-likehood (ML) tree based on 16S rRNA gene sequences showing the phylogenetic position of Deltaproteobacteria enriched from marine sediment (as inoculum) using different electron donors and sulfate sources (sodium sulfate or phosphogypsum). Representative sequences in the tree were obtained from GenBank (accession number in the brackets). Bootstrap values >75% are indicated at nodes. The bars represent the relative abundance of each OTU affiliated with Deltaproteobacteria in the enrichment cultures. The blue bars indicate the relative abundance of OTUs in sodium sulfate cultures, whereas the red bars represent the relative abundance of OTUs in PG cultures.
Figure 5 Maximum-likehood (ML) tree based on 16S rRNA gene sequences showing the phylogenetic position of Deltaproteobacteria enriched from marine sediment (as inoculum) using different electron donors and sulfate sources (sodium sulfate or phosphogypsum). Representative sequences in the tree were obtained from GenBank (accession number in the brackets). Bootstrap values >75% are indicated at nodes. The bars represent the relative abundance of each OTU affiliated with Deltaproteobacteria in the enrichment cultures. The blue bars indicate the relative abundance of OTUs in sodium sulfate cultures, whereas the red bars represent the relative abundance of OTUs in PG cultures.CC BY · PubMed Central
Figure 6 Maximum-likehood (ML) tree based on 16S rRNA gene sequences showing the phylogenetic position of Firmicutes from marine sediment (as inoculum) using different electron donors and sulfate sources (sodium sulfate or phosphogypsum). Representative sequences in the tree were obtained from GenBank (accession number in the brackets). Bootstrap values >75% are indicated at nodes. The bars represent the relative abundance (in %) of each OTU affiliated with Firmicutes in the enrichment cultures. The blue bars indicate the relative abundance of OTUs in sodium sulfate cultures, whereas the red bars represent the relative abundance of OTUs in PG cultures.
Figure 6 Maximum-likehood (ML) tree based on 16S rRNA gene sequences showing the phylogenetic position of Firmicutes from marine sediment (as inoculum) using different electron donors and sulfate sources (sodium sulfate or phosphogypsum). Representative sequences in the tree were obtained from GenBank (accession number in the brackets). Bootstrap values >75% are indicated at nodes. The bars represent the relative abundance (in %) of each OTU affiliated with Firmicutes in the enrichment cultures. The blue bars indicate the relative abundance of OTUs in sodium sulfate cultures, whereas the red bars represent the relative abundance of OTUs in PG cultures.CC BY · PubMed Central

Verification

Claims the paper makes

57 Own result · 22 Own interpretation · 17 No full text of the sources · 17 Not checked yet · 4 No basis given · 3 Own method · 2 Sources not found · 1 Supported · 1 Supported so far · 1 Partly supported · accuracy not yet measured

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Show all 87 claims

  • Probably not a substantial claim. 22
  • It may state two separate claims as one. 24
  • The quote only partly states this claim. 1
38 of 38
  • The paper relies on this without support.

    Discharge of PG into seawater or long-term PG stockpiling in close proximity to the sea presents a potential threat to surrounding coastal and marine ecosystems with substantial impact on human and environmental health and activities.

    “Moreover, the discharge of PG into the seawater or long-term PG stockpiling in close proximity to the sea present a potential threat to the surrounding coastal and marine ecosystems with substantial impact on human and environmental health and activities.”
  • The paper relies on this without support.

    The composition of total indigenous microbial populations inhabiting Tunisian PG has not yet been reported.

    “However, to the best of our knowledge, the composition of total indigenous microbial populations inhabiting Tunisian PG has not yet been reported.”
  • The paper relies on this without support.

    The biotechnological potential and diversity of marine SRB originating from PG-polluted coastal marine ecosystems to carry out such reductive process have not yet been studied.

    “Despite the abundance of SRB in marine ecosystems and their efficiencies in PG biotransformation, the biotechnological potential, and diversity of marine SRB, originating from PG-polluted coastal marine ecosystems, to carry out such reductive process have not yet been studied.”
  • The paper relies on this without support.

    Commercial development of PG biotransformation by SRB has been substantially limited by challenges including efficient reduction or immobilization of co-contaminants, use of low-cost and abundant renewable water sources for sulfate reduction, and improvement of sulfate reduction yields.

    “Thus, despite some advances in research on PG biotransformation by SRB (bioremediation process), its commercial development has been substantially limited by certain major challenges. These include the efficient reduction or immobilization of co-contaminants (i.e., trace metals, radionuclides), the use of low-cost and abundant renewable water sources for sulfate reduction (e.g., wastewaters and/or”
  • The paper relies on this without support.

    PG contains trace metals.

    “The absence of Desulfobacter spp. in PG cultures could be explained by the presence of trace metals in PG.”
  • The paper relies on this without support.

    Desulfovibrio spp. are powerful hydrogenotrophic SRB.

    “The first deltaproteobacterial group present in PG cultures comprised Desulfovibrio spp., which are known as powerful hydrogenotrophic SRB.”
  • The paper relies on this without support.

    Desulfobacter spp. are active AC-oxidizing SRB.

    “The second deltaproteobacterial group containing Desulfobacter spp. (Figure 5), which are active AC-oxidizing SRB (contrary to Desulfovibrio spp.), was only retrieved in Na2SO4 cultures, suggesting that such members were inhibited by PG.”
7 of 7

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Citations

Cites

Show all 62 references

Research works printed but not linked: 3

  1. Ben Amor, 2012, Reforme Politique Concernant la Gestion du Phosphogypse en Tunisie
  2. Gennari, 2011, Phosphogypsum analysis: total content and extractable element concentrations, 2011
  3. Inhibition of sulphate-reducing bacteria by metal sulphide formation in bioremediation of acid mine drainage4048 UtgikarV. P. HarmonS. M. ChaudharyN. TabakH. H. GovingR. HainesJ. R. 10.1002/tox.10031Environ. Toxicol.172002 · DOI

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Metadata

Authors
Zouch, H., Karray, F., Armougom, F., Chifflet, S., Hirschler-Réa, A., Kharrat, H., Kamoun, L., Ben Hania, W., Ollivier, B., Sayadi, S., & Quéméneur, M.
Affiliations
  • Centre National de la Recherche Scientifique (FR)
  • Centre of Biotechnology of Sfax (TN)
  • Groupe Chimique Tunisien (Tunisia) (TN)
Corresponding author
Quéméneur, Marianne
Venue
Frontiers in Microbiology
Published
2017-08-21
Volume
8
Pages
1583-1583
ISSN
1664-302X (ISSN-L)
OpenAlex
W2746696712
Cited by
37
References
62
Citations by year
2026: 2 · 2025: 3 · 2024: 2 · 2023: 6 · 2022: 2 · 2021: 8 · 2020: 4 · 2019: 8 · 2018: 2
FWCI
1.95
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Frontiers in Microbiology (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • HAL AMU (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Greater South Information System
Funders
Centre National de la Recherche Scientifique (EC2CO)
Topics
  • Physical Sciences › Environmental Science › Environmental Chemistry › Mine drainage and remediation techniques
  • Physical Sciences › Environmental Science › Ecology › Microbial Community Ecology and Physiology
  • Physical Sciences › Environmental Science › Environmental Engineering › Microbial Fuel Cells and Bioremediation
Keywords
phosphogypsum, sulfate-reducing bacteria, marine sediment, enrichment culture, sulfide production, Desulfovibrio, Desulfobacter, Deltaproteobacteria, electron donors, formate, bioremediation, metal immobilization, microbial diversity

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