The Paenibacillus polymyxa species is abundant among hydrogen-producing facultative anaerobic bacteria in Lake Averno sediment.

Lake Averno sediment was used to isolate the facultative anaerobic bacteria having the potential for H(2) production. Twenty-five out of 35 isolates recovered from the sediment sample produced hydrogen under anaerobic conditions from glucose with yields ranging from 0.1 to 0.49 mol H(2)/mol glucose. Identification based on 16S rRNA gene sequence analysis revealed that most of them belong to the Firmicutes group, with a prevalence of the Paenibacillus polymyxa species. Seven distinct genomic fingerprints among the 11 P. polymyxa isolates were obtained using the random amplified polymorphic DNA (RAPD) technique. Glucose fermentation by P. polymyxa isolates was investigated. Glucose was totally consumed after 3 days of fermentation. The fermentation products were hydrogen (0.18-0.47 mol H(2)/mol glucose), ethanol (0.1-0.5 mol ethanol/mol glucose), and 2,3-butanediol (0.1 mol 2,3-butanediol/mol glucose). Lower amounts of acetic, butyric, formic, lactic, and propionic acids were detected. All metabolic data concerning P. polymyxa isolates were analyzed by cluster analysis to reveal similarities and/or differences with clustering based on RAPD profiles. Despite the high metabolic similarity among almost all P. paenibacillus isolates, results of cluster analyses of metabolic and genetic data do not match completely.

Effects of Ciprofloxacin Alone or in Mixture with Sulfamethoxazole on the Efficiency of Anaerobic Digestion and Its Microbial Community.

Some livestock farms rely on anaerobic digestion (AD) technology for manure disposal, thus obtaining energy (biogas) and fertilizer (digestate). Mixtures of antibiotics used for animal health often occur in organic waste and their possible synergistic/antagonistic effects on microorganisms involved in AD are still poorly studied. This work focuses on the effects of adding ciprofloxacin, alone (5 mg L-1) and in combination with sulfamethoxazole (2.5-5-10 mg L-1), on AD efficiency and microbial community structure. The experiment consisted of 90-day cattle manure batch tests and antibiotic removal percentages were assessed. Adding antibiotics always promoted CH4 and H2 production compared to untreated controls; however, CH4 production was lowered with the highest ciprofloxacin (CIP) concentrations. The overall results show antibiotic degradation caused by acidogenic Bacteria, and CH4 was mainly produced through the hydrogenotrophic-pathway by methanogenic Archaea. Shifts in microbial community abundance (DAPI counts) and composition (Illumina-MiSeq and FISH analyses) were observed.

Bioaugmentation strategy to enhance polycyclic aromatic hydrocarbons anaerobic biodegradation in contaminated soils.

This paper proposes an innovative bioaugmentation approach for the remediation of polycyclic aromatic hydrocarbon (PAH) contaminated soils, based on a novel habitat-based strategy. This approach was tested using two inocula (i-24 and i-96) previously enriched through an anaerobic digestion process on wheat straw. It relies on the application of allochthonous microorganisms characterized by specific functional roles obtained by mimicking a natural hydrolytic environment such as the rumen. The inocula efficiency was tested in presence of naphthalene alone, benzo[a]pyrene alone, and a mix of both of them. In single-contamination tests, i-24 inoculum showed the highest biodegradation rates (84.7% for naphthalene and 51.7% for benzo[a]pyrene). These values were almost 1.2 times higher than those obtained for both contaminants with i-96 inoculum and in the control test in presence of naphthalene alone, while they were 3 times higher compared to the control test in presence of benzo[a]pyrene alone. In mixed-contamination tests, i-96 inoculum showed final biodegradation efficiencies for naphthalene and benzo[a]pyrene between 1.1 and 1.5 higher than i-24 inoculum or autochthonous biomass. Total microbial abundances increased in the bioaugmented tests in line with the PAH degradation. The microbial community structure showed the highest diversity at the end of the experiment in almost all cases. Values of the Firmicutes active fraction up to 7 times lower were observed in the i-24 bioaugmented tests compared to i-96 and control tests. This study highlights a successful bioaugmentation strategy with biological components that can be reused in multiple processes supporting an integrated and environmentally sustainable bioremediation system.

A novel enrichment approach for anaerobic digestion of lignocellulosic biomass: Process performance enhancement through an inoculum habitat selection.

Inocula enrichment was performed using an innovative habitat-based selection approach to improve wheat straw (WS) anaerobic digestion (AD) efficiency. The procedure was carried out by sequentially re-inoculating the primary microbial community seven times in subsequent anaerobic reactors containing untreated WS. Re-inocula were performed at different re-inoculum times (24, 48, and 96 h) by moving a porous support mimicking a rumen structure from one batch to the next (S-tests) or re-inoculating only the culture medium (C-tests). Highest H2 production yields were observed after four and five re-inocula (0.08 ± 0.02 NmL h-1 gVS-1 and 0.09 ± 0.02 NmL h-1 gVS-1) for S-24 and S-48, respectively. For S-96, higher CH4 yields were observed after the start-up test and sixth re-inoculum (0.05 ± 0.003 NmL h-1 gVS-1 and 0.04 ± 0.005 NmL h-1 gVS-1, respectively). Accordingly, S-96 showed the highest active Archaea component (7%). C-test microbial communities were dominated by fermenting, hydrogen-producing bacteria and showed lower microbial community diversity than S-tests.

Effects of Sulfamethoxazole on the Microbial Community Dynamics During the Anaerobic Digestion Process.

Anaerobic digestion (AD) treatment of cattle manure and slurry makes it possible to produce biogas, a renewable and storable biofuel, as well as digestate, a residual organic matter that can be used to replace chemical fertilizers. On the other hand, the intense use of antibiotics (e.g., sulfamethoxazole) in animal husbandry practices is showing increasing negative impacts resulting from the release of still metabolically active molecules into agroecosystems. In the present study, cattle manure collected from an AD plant-feeding tank was used as feedstock for AD experiments in which some batches were spiked with 5 mg L-1 of sulfamethoxazole (SMX). Adding the antibiotic affected the microbial community dynamic; in particular, the efficiency of the acidogenic and acetogenic phases of the process corresponded to higher CH4 and H2 production than in the control. SMX was also degraded, and at the end of the experiment (69 days), just 20% of its initial concentration was found. The relative abundance (ARG/16S) of resistance genes sul1, sul2, and the proxy intI1 initially found in the ingestate decreased during the AD in both the spiked and control batches, suggesting that this process lowers the likelihood of antibiotic resistance genes spreading.

A simplified model to simulate bioaugmented anaerobic digestion of lignocellulosic biomass: Biogas production efficiency related to microbiological data.

Mathematical model applications for the bioaugmented anaerobic digestion (BAD) process seem to be lacking in the scientific literature, even more so when related to microbiological data. The present study suggests a simplified mathematical model to investigate and simulate the process kinetics of bioaugmented anaerobic digestion (BAD) aimed at improving biogas production from wheat straw (WS). Bioaugmented conditions were obtained through a mixed inoculum of anaerobic ruminal fungi (ARF) and hydrogen-producing fermenting bacteria (F210) added to a methanogenic inoculum. The investigation focused on two process configurations characterized by a mono (I-BAD) and two-stage (II-BAD) process and a conventional anaerobic digestion (AD) control test. Each configuration was used on two operating scales (i.e., 120 ml and 12,000 ml reactor volume) to provide different data sets for the calibration and validation of the mathematical model proposed. The model calibration step was used to determine the optimal values of selected parameters displaying higher significance for experimental result predictability. The model calibration results highlighted a similar behavior for both BAD tests, which was further strengthened by a statistical analysis supporting the observed correlation regardless of the BAD configuration involved. The BAD configuration always enhanced the CH4 production (>70%) with a faster kinetic in the II-BAD test. The microbiological results support the superior performance of the II-BAD test, displaying higher Archaea fractions (up to 14.5% on day 23) with values more than double compared to I-BAD and triple compared to the AD test. Furthermore, the presence of Methanosarcina inside the Archaea guild (6.4% and 4.5% at days 11 and 61, respectively) ensures a greater diversification of the metabolic pathways and supports the strength of the process performance. Cell density values are strongly in line with these results.

Combined bioaugmentation with anaerobic ruminal fungi and fermentative bacteria to enhance biogas production from wheat straw and mushroom spent straw.

Bioaugmentation with anaerobic ruminal fungi and a pool of hydrogen-producing fermenting bacteria was tested on wheat straw (WS) and mushroom spent straw (MSS) with the aim of improving anaerobic digestion performance. Batch tests were set up to simulate a Bioaugmentation Anaerobic Digestion (BAD) treatment comparing single- (I-BAD) and two-stage (II-BAD) process configurations, at two reactor scales, 120 and 1200 ml (×10). In both cases, higher CH4 cumulative production was obtained in the II-BAD configuration on WS (65.1 ±â€¯8.9 Nml and 922 ±â€¯73.8 Nml respectively). The II-BADx10 tests allowed increasing CH4 production (≃290% and ≃330% on WS and MSS, respectively) when compared to the unaugmented condition. Final results highlighted the achievable advantages of the two stage configuration in terms of CH4 production enhancement. Microbial community investigations confirmed the efficiency of the bioaugmentation treatment and revealed that such a result was mainly related to the Methanosarcinales increase, mostly composed by Methanosaeta.

Vertical distribution of bacterioplankton in Lake Averno in relation to water chemistry.

The effects of environmental factors on bacterioplankton distribution along the water column of Lake Averno (Naples, Italy) have been investigated by means of denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR)-amplified 16S rRNA gene fragments, and multivariate analysis applied to molecular data and physico-chemical parameters. Bacterial richness, estimated from DGGE profiles, remains constant throughout the water column, whereas archaeal richness increases with depth. Moreover, archaeal richness was significantly correlated to most of the measured abiotic variables, whereas bacterial richness did not. Analysis of sequences from DGGE bands revealed that the dominant bacterial populations belong to Alphaproteobacteria, Betaproteobacteria, Deltaproteobacteria, Bacteroidetes and Firmicutes, whereas sequences of the archaeal DGGE bands are affiliated to Methanomicrobiales and Methanosarcinales orders. Moreover, bacterial sequences affiliated to Proteobacteria and Bacteroidetes do not group closely to typical freshwater lineages/clades/tribes within these phyla. Bacterioplankton distribution along the water column was apparently correlated with the vertical gradient of physico-chemical parameters as a statistically significant relationship between most of them with grouping of specific taxonomic units was observed.