Effects of the operational conditions on the production of 1,3-propanediol derived from glycerol in anaerobic granular sludge reactors.

The aim of this study has been to produce 1,3-propanediol (1,3-PDO) from glycerol (gly) fermentation by means of a microbial mixed culture (granular sludge), as well as to establish the operational conditions of two up-flow anaerobic sludge blanket (UASB) reactors in order to achieve a maximum 1,3-PDO yield. The UASB reactors with initial pH values set at 6.8 and 5.5 were operated at 30 °C during 165 days. Thirteen variables were previously screened by a Plackett-Burman (PB) design; results showed that yeast extract, MgSO4 and methanogenesis inhibition (by heat shock) showed a positive effect, whereas high glycerol concentration, tryptone and CaCl2 showed a negative impact on the 1,3-PDO produced by glycerol degradation. Following four experimental periods, the highest average yield of 0.43 mol 1,3-PDO mol-1 gly was achieved when sodium bicarbonate was added to the reactors. Propionate and acetate were also produced and a high microorganism diversity was detected; however, the restrictive operational conditions of the reactors led to the death of the methanogenic archaea. Nevertheless, the continuous production of 1,3-PDO from glycerol within UASB reactors inoculated with granular sludge can be considered highly feasible.

Proteolytic bacterial dominance in a full-scale municipal solid waste anaerobic reactor assessed by 454 pyrosequencing technology.

Biomethanization entails a good means to reduce the organic fraction (OF) derived from municipal solid wastes (MSW). The bacterial diversity of a full scale MSW anaerobic reactor located in Madrid (Spain) was investigated using high-throughput 454 pyrosequencing. Even though the proteolytic bacteria prevailed throughout all of the process, community shifts were observed from the start-up to the steady-state conditions, with an increasing biodiversity displayed over time. The Bacteroidetes and the Firmicutes were the majority phyla: 55.1 and 40.2% (start-up) and 18.7 and 78.7 (steady-state) of the total reads. The system's lack of evenness remains noteworthy as the sequences affiliated to the proteolytic non-saccharolytic Proteiniphylum, Gallicola and Fastidiosipila genera, together with the saccharolytic Saccharofermentans, were predominant on the system and this predominance appears to correlate with the presence of a high ammonium concentration. The 454 pyrosequencing revealed a great diversity of rare organisms which seemingly do not sustain any metabolic roles in the course of the OF-MSW degradation. However, this scarce and unique microbiota can confer great resilience to the system as a buffer against nutritional and environmental changing conditions, thus opening the door to increase the current knowledge about the bacterial community dynamics taking place during MSW treatment processes.

Organic loading rate and food-to-microorganism ratio shape prokaryotic diversity in a demo-scale up-flow anaerobic sludge blanket reactor treating domestic wastewater.

We investigated the microbial community in an up-flow anaerobic sludge blanket (UASB) reactor treating domestic wastewater (DW) during two different periods of organic loading rate (OLR) and food-to-microorganism (F/M) ratio. 16S rDNA clone libraries were generated, and quantitative real-time PCR (qPCR) analyses were performed. Fluctuations in the OLR and F/M ratio affected the abundance and the composition of the UASB prokaryotic community, mainly at the species level, as well as the performance of the UASB reactor. The qPCR analysis suggested that there was a decrease in the bacterial cell number during the rainy season, when the OLR and F/M ratio were lower. However, the bacterial diversity was higher during this time, suggesting that the community degraded more diversified substrates. The diversity and the abundance of the archaeal community were higher when the F/M ratio was lower. Shifts in the methanogenic community composition might have influenced the route of methane production, with methane produced by acetotrophic methanogens (dry season), and by hydrogenotrophic, methylotrophic and acetotrophic methanogens (rainy season). This study revealed higher levels of bacterial diversity, metabolic specialization and chemical oxygen demand removal efficiency of the DW UASB reactor during the rainy season.

Prokaryotic diversity and dynamics in a full-scale municipal solid waste anaerobic reactor from start-up to steady-state conditions.

The prokaryotic diversity of an anaerobic reactor for the treatment of municipal solid waste was investigated over the course of 2 years with the use of 16S rDNA-targeted molecular approaches. The fermentative Bacteroidetes and Firmicutes predominated, and Proteobacteria, Actinobacteria, Tenericutes and the candidate division WWE1 were also identified. Methane production was dominated by the hydrogenotrophic Methanomicrobiales (Methanoculleus sp.) and their syntrophic association with acetate-utilizing and propionate-oxidizing bacteria. qPCR demonstrated the predominance of the hydrogenotrophic over aceticlastic Methanosarcinaceae (Methanosarcina sp. and Methanimicrococcus sp.), and Methanosaetaceae (Methanosaeta sp.) were measured in low numbers in the reactor. According to the FISH and CARD-FISH analyses, Bacteria and Archaea accounted for 85% and 15% of the cells, respectively. Different cell counts for these domains were obtained by qPCR versus FISH analyses. The use of several molecular tools increases our knowledge of the prokaryotic community dynamics from start-up to steady-state conditions in a full-scale MSW reactor.

Bacterial and archaeal phylogenetic diversity associated with swine sludge from an anaerobic treatment lagoon.

Over the last decades, the demand for pork products has increased significantly, along with concern about suitable waste management. Anaerobic-lagoon fermentation for swine-sludge stabilization is a good strategy, although little is known about the microbial communities in the lagoons. Here, we employed a cloning- and sequencing-based analysis of the 16S rRNA gene to characterize and quantify the prokaryotic community composition in a swine-waste-sludge anaerobic lagoon (SAL). DNA sequence analysis revealed that the SAL library harbored 15 bacterial phyla: Bacteroidetes, Cloroflexi, Proteobacteria, Firmicutes, Deinococcus-Thermus, Synergystetes, Gemmatimonadetes, Chlorobi, Fibrobacteres, Verrucomicrobia and candidates division OP5, OP8, WWE1, KSB1, WS6. The SAL library was generally dominated by carbohydrate-oxidizing bacteria. The archaeal sequences were related to the Crenarchaeota and Euryarchaeota phyla. Crenarchaeota predominated in the library, demonstrating that it is not restricted to high-temperature environments, being also responsible for ammonium oxidation in the anaerobic lagoon. Euryarchaeota sequences were associated with the hydrogenotrophic methanogens (Methanomicrobiales and Methanobacteriales). Quantitative PCR analysis revealed that the number of bacterial cells was at least three orders of magnitude higher than the number of archaeal cells in the SAL. The identified prokaryotic diversity was ecologically significant, particularly the archaeal community of hydrogenotrophic methanogens, which was responsible for methane production in the anaerobic lagoon. This study provided insight into the archaeal involvement in the overall oxidation of organic matter and the production of methane. Therefore, the treatment of swine waste in the sludge anaerobic lagoon could represent a potential inoculum for the start-up of municipal solid-waste digesters.

Remoción de percloroetileno en dos tipos de sistemas anaerobios continuos

Se evaluó y comparó el desempeño de dos reactores mesofílicos metanogénicos de mezcla completa escala laboratorio (RMC1, RMC2) y un reactor metanogénico de lecho fluidizado (RANLEF, con carbón activado granular como soporte) para remoción de percloretileno (PCE) tras adicionar una moderada cantidad de metanol en el afluente. Los reactores fueron operados con cargas de 0,07 y 1gDQO·l-1·d-1, y tiempos de retención hidráulica de 15 y 1d, respectivamente. El diseño experimental consistió en tres períodos de operación: régimen metanogénico con metanol como fuente de C (periodo 1), igual con aclimatación a 20mgPCE·l-1 (periodo 2.1) o 40mgPCE·l-1 (2.2), e igual con 40mgPCE·l-1 en RANLEF y RMC1, y 20mgPCE·l-1 en RMC2 (periodo 3). La eficiencia de PCE y DQO fue más alta en RANLEF en el primer periodo; el CH4 en biogás y el incremento de cloro fueron similares en los tres reactores. Durante el periodo 2.2 la remoción de PCE alcanzó 81% en RANLEF, sugestivo de buena aclimatación a mayor entrada de PCE; la remoción de DQO se mantuvo alta, y el contenido de CH4 en biogás alcanzó un valor medio de 64% v/v, indicando un régimen metanogénico satisfactorio. Los RMCs experimentaron un deterioro drástico en su desempeño al aumentar PCE de 20 a 40mg·l-1, por lo que se regresó el RMC1 a 20mg·l-1. El desempeño de RMC2 se mantuvo bajo, sugiriendo un impacto negativo sobre la metanogénesis y remoción de PCE. En contraste, el RMC1 con 20mg·l-1 se recuperó del estado transitorio negativo, exhibiendo desempeño similar al RANLEF alimentado con 40mg·l-1. En el período 3, el RANLEF con 40mg·l-1 y el RMC1 con 20mg·l-1 mostraron desempeño similar, mientras que el RMC2 con 40mg·l-1 exhibió una disminución en la eficiencia de remoción de DQO, baja productividad de biogás y contenido de CH4 en biogás, pero la remoción de PCE estuvo cerca de RANLEF y RMC1. La eficiencia de descloración del RMC2 fue significativamente menor que la del RANLEF, el cual fue más robusto y estable en el proceso anaerobio de remoción de alta concentración de PCE.

Performance was evaluated and compared in two lab-scale mesophilic methanogenic complete mix reactors (RMC1, RMC2) and a methanogenic fluidized bed reactor (RANLEF) for removal of perchloroethylene (PCE) when fed a moderate concentration of methanol. The RMCs and RANLEF were operated at loading rates of 0.07 and 1gCOD·l-1·d-1, and hydraulic retention times of 15 and 1 day, respectively. The experimental design consisted of three periods of operation: methanogenic regime with methanol as carbon source (1); same with acclimation to 20 (2.1) or 40mgPCE·l-1 (2.2); and same with 40mgPCE·l-1 in RANLEF and RMC1 and 20 mgPCE·l-1 in RMC2 (3). In the first period both PCE and COD removals were higher in RANLEF; biogas CH4 content and chloride increase were similar in the three reactors. During period 2.2, PCE removal increased up to 81% in RANLEF, suggesting a good acclimation to the higher inflow PCE concentration; COD reduction remained high, and CH4 in biogas indicated a satisfactory methanogenic regime. The RMCs experienced drastic performance impairments upon increase of inflow PCE (20 to 40mg·l-1), and RMC1 was returned to operate with 20mgPCE·l-1. RMC2 performance remained poor, showing a drastic deterioration of methanogenesis and PCE removal. By contrast, RMC1 with 20mgPCE·l-1 recovered from the negative transient state and exhibited similar performance to that of RANLEF fed 40mg·l-1. In period 3, RANLEF with 40mg·l-1 and RMC1 with 20mg·l-1 showed similar performances, whereas RMC2 kept with 40mg·l-1 exhibited impaired COD removal, lower biogas productivity and CH4 biogas content, but a PCE removal very close to RANLEF and RMC1. The dehalogenation efficiency of RMC2 was significantly lower than that of the RANLEF, which appears to be a more robust and stable anaerobic process for the effective removal of high concentrations of PCE.

Avaliou-se e comparou-se o desempenho de dois reatores mesofílicos metanogênicos de mistura completa escala laboratório (RMC1, RMC2) e um reator metanogênico de leito fluidizado (RANLEF, com carvão ativado granular como suporte) para remoção de percloretileno (PCE) depois de adicionar uma moderada quantidade de metanol no afluente. Os reatores foram operados com cargas de 0,07 e 1gDQO·l-1·d-1, e tempos de retenção hidráulica de 15 e 1d, respectivamente. O projeto experimental consistiu em três períodos de operação: regime metanogênico com CH4 como fonte de carbono (1), igual com aclimatação a 20mgPCE·l-1 (2.1) ou 40mgPCE·l-1 (2.2), e igual com 40mgPCE·l-1 em RANLEF e RMC1, e 20mgPCE·l-1 em RMC2 (3). A eficiência de PCE e DQO foi mais alta em RANLEF no primeiro período; o CH4 em biogás e o incremento de cloro foram similares nos três reatores. Durante o período 2.2 a remoção de PCE alcançou 81% em RANLEF, sugestivo de boa aclimatação a maior entrada de PCE; a remoção de DQO se manteve alta, e o conteúdo de CH4 em biogás alcançou um valor médio de 64% v/v, indicando um regime metanogênico satisfatório. Os RMCs experimentaram um deterioro drástico no seu desempenho ao aumentar PCE de 20 a 40mg·l-1, pelo que se regressou o RMC1 a 20mg·l-1. O desempenho de RMC2 se manteve baixo, sugerindo um impacto negativo sobre a metanogênesis e remoção de PCE. Em contraste, o RMC1 com 20mg·l-1 se recuperou do estado transitório negativo, exibindo desempenho similar ao RANLEF alimentado com 40mg·l-1. No período 3, o RANLEF com 40mg·l-1 e o RMC1 com 20mg·l-1 mostraram desempenho similar, enquanto que o RMC2 com 40mg·l-1 exibiu uma diminuição na eficiência de remoção de DQO, baixa produtividade de biogás e conteúdo de CH4 em biogás, mas a remoção de PCE esteve perto de RANLEF e RMC1. A eficiência de descloração do RMC2 foi significativamente menor que a do RANLEF, o qual foi mais robusto e estável no processo anaeróbio de remoção de alta concentração de PCE.