2,4-Dichlorophenoxyacetic acid degradation in methanogenic mixed cultures obtained from Brazilian Amazonian soil samples.

2,4-Dichlorophenoxyacetic acid (2,4-D) is the third most applied pesticide in Brazil to control broadleaf weeds in crop cultivation and pastures. Due to 2,4-D's high mobility and long half-life under anoxic conditions, this herbicide has high probability for groundwater contamination. Bioremediation is an attractive solution for 2,4-D contaminated anoxic environments, but there is limited understanding of anaerobic 2,4-D biodegradation. In this study, methanogenic enrichment cultures were obtained from Amazonian top soil (0-40 cm) and deep soil (50 -80 cm below ground) that biotransform 2,4-D (5 µM) to 4-chlorophenol and phenol. When these cultures were transferred (10% v/v) to fresh medium containing 40 µM or 160 µM 2,4-D, the rate of 2,4-D degradation decreased, and biotransformation did not proceed beyond 4-chlorophenol and 2,4-dichlorophenol in the top and deep soil cultures, respectively. 16S rRNA gene sequencing and qPCR of a selection of microbes revealed no significant enrichment of known organohalide-respiring bacteria. Furthermore, a member of the genus Cryptanaerobacter was identified as possibly responsible for phenol conversion to benzoate in the top soil inoculated culture. Overall, these results demonstrate the effect of 2,4-D concentration on biodegradation and microbial community composition, which are both important factors when developing pesticide bioremediation technologies.

Heavy metal concentrations in Brazilian port areas and their relationships with microorganisms: can pollution in these areas change the microbial community?

The objectives of this study were to analyze the difference in ways in which metals polluting Brazilian port areas influence bacterial communities and the selection of resistant strains. The hypothesis tested was that port areas would have microbial communities significantly different from a pristine area, mainly due to a greater load of metals found in these areas. Sediment samples were collected in two port areas (Santos and São Sebastião) and one pristine area (Ubatuba). Total DNA was extracted and MiSeq sequencing was performed. A hundred strains were isolated from the same samples and were tested for metal resistance. The community composition was similar in the two port regions, but differed from the pristine area. Microbial diversity was significantly lower in the port areas. The phyla Proteobacteria, Cyanobacteria, and Thermodesulfobacteria exhibited positive correlations with copper and zinc concentrations. Chloroflex, Nitrospirae, Planctomycetes, and Chlorobi exhibited negative correlations with copper, chromium, and zinc. Cr and Zn had higher concentrations at port areas and were responsible to select more metal-resistant strains. Some genera were found to be able to easily develop metal resistance. The most isolated genera were Bacillus, Vibrio, and Pseudomonas. This type of study can illustrate, even in very complex natural environments, the influence of pollution on the community as a whole and the consequences of these changes.

Biofilm stratification and autotrophic-heterotrophic interactions in a structured bed reactor (SBRIA) for carbon and nitrogen removal.

The structured-bed reactor with intermittent aeration (SBRIA) is a promising technology for simultaneous carbon and nitrogen removal from wastewater. An in depth understanding of the microbiological in the reactor is crucial for its optimization. In this research, biofilm samples from the aerobic and anoxic zones of an SBRIA were analyzed through 16S rRNA sequencing to evaluate the bacterial community shift with variations in the airflow and aeration time. The control of the airflow and aeration time were essential to guarantee reactor performances to nitrogen removal close to 80%, as it interfered in nitrifying and denitrifying communities. The aeration time of 1.75 h led to establishment of different nitrogen removal pathways by syntrophic relationships between nitrifier, denitrifier and anammox species. Additionally, the predominance of these different species in the internal and external parts of the biofilm varied according to the airflow.

Lactate as an effective electron donor in the sulfate reduction: impacts on the microbial diversity.

The competition between sulfate-reducing bacteria and methane-producing archaea has a major influence on organic matter removal, as well as the success of sulfidogenic systems. This study investigated the performance of six batch sulfidogenic reactors in response to different COD/sulfate ratios (1.0 and 2.0) and electron donors (cheese whey, ethanol, and sodium lactate) by evaluating the biochemical mechanisms of sulfate reduction, organic matter oxidation, and microbial structure modification. A COD/sulfate ratio of 1.0 resulted in high sulfidogenic activity for all electron donors, thereby achieving a nearly 80% sulfate removal. Lactate provided high sulfate removal rates at COD/sulfate ratios of 1.0 (80%) and 2.0 (90%). A COD/sulfate ratio of 2.0 decreased the sulfate removal rates by 25 and 28% when ethanol and cheese whey were used as substrates. The sulfate-reducing bacteria populations increased using ethanol and lactate at a COD/sulfate ratio of 1.0. Particularly, Desulfovibrio, Clostridium, and Syntrophobacter were predominant. Influent composition and COD/sulfate ratio influenced the relative abundance of the microbial communities. Therefore, controlling these parameters may facilitate the wastewater treatment with high sulfate levels through bacterial activity.

Effects of metals on activity and community of sulfate-reducing bacterial enrichments and the discovery of a new heavy metal-resistant SRB from Santos Port sediment (São Paulo, Brazil).

Sulfate-reducing bacteria (SRB) can be used to remove metals from wastewater, sewage, and contaminated areas. However, metals can be toxic to this group of bacteria. Sediments from port areas present abundance of SRB and also metal contamination. Their microbial community has been exposed to metals and can be a good inoculum for isolation of metal-resistant SRB. The objective of the study was to analyze how metals influence activity and composition of sulfate-reducing bacteria. Enrichment cultures were prepared with a different metal (Zn, Cr, Cu, and Cd) range concentration tracking activity of SRB and 16S rRNA sequencing in order to access the community. The SRB activity decreased when there was an increase in the concentration of the metals tested. The highest concentration of metals precipitated were 0.2 mM of Cd, 5.4 mM of Zn, 4.5 mM of Cu, and 9.6 mM of Cr. The more toxic metals were Cd and Cu and had a greater community similarity with less SRB and more fermenters (e.g., Citrobacter and Clostridium). Meanwhile, the enrichments with less toxic metals (Cr and Zn) had more sequences affiliated to SRB genera (mainly Desulfovibrio). A new Desulfovibrio species was isolated. This type of study can be useful to understand the effects of metals in SRB communities and help to optimize wastewater treatment processes contaminated by metals. The new Desulfovibrio species may be important in future studies on bioremediation of neutral pH effluents contaminated by metals.

Sulfate and metal removal from acid mine drainage using sugarcane vinasse as electron donor: Performance and microbial community of the down-flow structured-bed bioreactor.

The down flow structured bed bioreactor (DFSBR) was applied to treat synthetic acid mine drainage (AMD) to reduce sulfate, increase the pH and precipitate metals in solutions (Co, Cu, Fe, Mn, Ni and Zn) using vinasse as an electron donor for sulfate-reducing bacteria (SRB). DFSBR achieved sulfate removal efficiencies between 55 and 91%, removal of Co and Ni were obtained with efficiencies greater than 80%, while Fe, Zn, Cu and Mn were removed with average efficiencies of 70, 80, 73 and 60%, respectively. Sulfate reduction increased pH from moderately acidic to 6.7-7.5. Modelling data confirmed the experimental results and metal sulfide precipitation was the mainly responsible for metal removal. The main genera responsible for sulfate and metal reduction were Geobacter and Desulfovibrio while fermenters were Parabacteroides and Sulfurovum. Moreover, in syntrophism with SRB, they played an important role in the efficiency of metal and sulfate removal.

Characterization of Alkaliphilic Bacteria Isolated from Bauxite Residue in the Southern Region of Minas Gerais, Brazil

ABSTRACT The aim of this study was to isolate and characterize bacterial strains from bauxite residue in the southern region of Minas Gerais, Brazil, by 16S rRNA gene sequencing and biochemical assays. Bacillus cohnii, Bacillus pseudofirmus, and Bacillus clarkii were identified among the isolates. The isolates were able to use a wide range of carbon sources and to grow at NaCl concentrations of up to 10%, temperatures from 10 to 40 °C, and pH from 7 to 10.5, producing a wide variety of organic acids. This is the first report on microbial composition of bauxite residue in Brazil.