Reduction in the in vitro ruminal methane production with the inclusion of Cucurbita foetidissima / Reducción en la producción ruminal de metano in vitro con la inclusión de Cucurbita foetidissima

ABSTRACT Objective. This research aimed to evaluate the inclusion of Cucurbita foetidissima (BG) leaves as a partial or total substitution of alfalfa hay (AH) in beef cattle diets on in vitro methane output, gas kinetics and volatile fatty acids production. Materials and Methods. Five experimental treatments were formulated with the inclusion of BG as alfalfa hay (AH) substitute at 0, 25, 50, 75 and 100% in experimental treatments denominated CON (control), BG25, BG50, BG75 and BG100, respectively. Results. Lignin and organic matter decreased with BG inclusion (p<0.05). However, NDF, CT and TPC increased with higher levels of BG (p<0.05). Maximum gas production diminished with BG inclusion (p<0.05); whereas, microbial protein production, specific gas production rate and latency period were not affected (p>0.05). Methane production decreased linearly with BG inclusion (p<0.05). Nevertheless, CO2 production showed no changes with inclusion of graded levels of BG in the experimental treatments (p>0.05). Additionally, acetate and butyrate were not affected by BG inclusion (p>0.05); although, propionate increased linearly among treatments (p<0.05). Furthermore, inclusion of 75% of BG reduced 31% in vitro methane production without changes in CO2 production, suggesting an inhibition of endogenous methanogenesis. Conclusions. These results promote the inclusion of BG leaves as a potential and environmentally friendly forage source for beef cattle feeding.

RESUMEN Objetivo. El objetivo de este estudio fue evaluar la inclusión de hojas de Cucurbita foetidissima (BG) como sustituto parcial o total de la alfafa (AH) en la dieta de ganado de engorda sobre la cinética de producción de gas y producción ruminal in vitro de metano y ácidos grasos volátiles. Materiales y Métodos. Se formularon cinco dietas con la inclusión de BG como sustituto de heno de alfalfa a 0, 25, 50, 75 y 100% en los tratamientos experimentales que se denominaron CON (control), BG25, BG50, BG75 y BG100, respectivamente. Resultados. La lignina y la materia orgánica disminuyeron con la inclusión de BG (p<0.05). La máxima producción de gas disminuyó con la inclusión de BG (p<0.05); mientras que la producción de proteína microbiana, la tasa específica de producción de gas y el tiempo de latencia no mostraron cambios(p>0.05). La producción de metano disminuyó linealmente con la inclusión de BG (p<0.05). No obstante, la producción de CO2 no mostró cambios con la inclusión de BG en los tratamientos (p<0.05). Adicionalmente, el acetato y el butirato no fueron afectados por la inclusión de BG (p>0.05); a pesar de que el propionato se incrementó linealmente (p<0.05). Además, la inclusión de BG en un 75% redujo la producción de metano 31% sin afectar la producción de CO2, lo que sugiere una inhibición de la metanogenesis endógena. Conclusiones. Estos resultados promueven la inclusión de hojas de BG como una fuente de forraje potencial y amigable con el ambiente en la alimentación de ganado de engorda.

Therapeutic effect of methane and its mechanism in disease treatment / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Methane is the simplest hydrocarbon, consisting of one carbon atom and four hydrogen atoms. It is abundant in marsh gas, livestock rumination, and combustible ice. Little is known about the use of methane in human disease treatment. Current research indicates that methane is useful for treating several diseases including ischemia and reperfusion injury, and inflammatory diseases. The mechanisms underlying the protective effects of methane appear primarily to involve anti-oxidation, anti-inflammation, and anti-apoptosis. In this review, we describe the beneficial effects of methane on different diseases, summarize possible mechanisms by which methane may act in these conditions, and discuss the purpose of methane production in hypoxic conditions. Then we propose several promising directions for the future research.

Anaerobic co-digestion of swine manure with sweet potato or cassava in different C/N ratios / Co-digestão anaeróbia dos dejetos de suínos com batata doce ou mandioca em diferentes relações C/N

ABSTRACT: Anaerobic co-digestion (AcoD) of waste is a method of increasing methane (CH4) yield and improving biofertilizer quality. This study aimed to evaluate the best AcoD conditions for swine manure (SM) with sweet potato (SP) or cassava (C) in different amounts in semi-continuous biodigesters. Initially, using batch biodigesters, an AcoD test of the SM with SP or C was performed, adopting carbon/nitrogen (C/N) ratios of 10/1, 13/1, 17/1, and 22/1. Based on the results, a C/N ratio of 10/1 was chosen, which was the proportion that resulted in the highest reduction of volatile solids (VS) and specific biogas production. From these results, the experiment was carried out in semi-continuous biodigesters, consisting of three treatments (control (SM), SP 10/1, and C 10/1) with five replicates and a hydraulic retention time (HRT) of 30 days. Total solid (TS) and volatile solid (VS) reductions, biogas and CH4 yields, alkalinity, and volatile acidity were measured. The control treatment differed from the others and resulted in decreased biogas yield (752 LN.kgVSadded -1), CH4 (449 LN.kgVSadded -1), and CH4 content (59.7%). The AcoD treatments (SP and C) did not differ significantly for biogas yield (respectively, 901 and 883 LN.kgVSadded -1) and CH4 (respectively, 590 and 547 LN.kgVSadded -1); however, they differed in CH4 content (65.5% and 61.9% respectively). The treatments showed general reduction averages of 76.1% and 85.9% for TS and VS, respectively, with no statistical difference found between them. The AcoD of the SM with SP or C increased the production and quality of the biogas, increasing the concentration of CH4 therein.

RESUMO: A co-digestão anaeróbia dos resíduos é uma alternativa para aumentar a produção de metano (CH4) e melhorar a qualidade do biofertilizante. Este estudo teve como objetivo avaliar a melhor condição de coDA dos dejetos de suínos (DS), acrescidos de batata doce (BD) ou mandioca (M) em diferentes inclusões, em biodigestores semi-contínuos. Inicialmente, utilizando biodigestores batelada, foi realizado ensaio de coDA dos DS com BD ou M, adotando-se as relações C/N iguais a 10/1, 13/1, 17/1, e 22/1. Com base nos resultados elegeu-se a relação C/N de 10/1, como sendo a proporção que apresentou maiores reduções de sólidos voláteis (SV) e produções específicas de biogás. A partir destes resultados, efetivou-se o experimento em biodigestores semi-contínuos, composto por três tratamentos (controle (DS), BD 10/1 e M 10/1) com cinco repetições cada, e tempo de retenção hidráulica (TRH) de 30 dias. Foram mensuradas as reduções de sólidos totais (ST) e sólidos voláteis (SV), produções de biogás e CH4, alcalinidade e acidez volátil. O tratamento controle diferiu dos demais e apresentou menor desempenho no rendimento de biogás (752 LN.kgSVadic -1), CH4 (449 LN.kgSVadic -1) e concentração de CH4 (59,7%). Os tratamentos em coDA (BD e M) não diferiram entre si para o rendimento de biogás (901 e 883 LN.kgSVadic -1) e CH4 (590 e 547 LN.kgSVadic -1), no entanto diferiram na concentração de CH4 (65,5 e 61,9%, respectivamente). Os tratamentos apresentaram médias gerais de reduções de 76,1 e 85,9% para ST e SV, respectivamente, não sendo encontrada diferença estatística entre eles. A coDA do DS com BD ou M aumentou a produção e a qualidade do biogás, elevando a concentração de CH4 no mesmo.

Population Changes of Eubacteria, Sulfate-Reducing Bacteria and Methanogenic Archaea in an Anaerobic Reactor Processing Ethanol Distillery Vinasse

Abstract The microbiological characterization by molecular techniques (DGGE and quantitative PCR) of Archaea, Bacteria domain and sulfate-reducing bacteria (SRB) from a laboratory scale Up-flow Anaerobic Sludge Blanket (UASB) processing sugar cane vinasse was performed during the operational phase with increasing organic loads. The organic load removal efficiency was between 97% and 75% for volumetric organic loads (VOL) in the range of 0.6 to 15.4 kgCOD.m-3.d−1 and for higher VOL (until 27.0 kgCOD.m-3 .d−1) the removal efficiency decreased to 48%. Archaea represented the majority of the estimated population (107 copies of 16S RNA ribosomal gene. mL-1) followed by bacteria (106 copies of 16S RNA ribosomal gene. mL-1) and sulfate-reducing bacteria (SRB) (105 copies of dsrB gene. mL-1). There was a significant difference between the populations of Archaea and SRB with increasing VOL. DGGE profiles show differences among populations with increasing VOL, especially with respect to sulfate-reducing bacteria. The presence of SRB together with a high sulfate removal percentage (97%) of about 15 kgCOD.m-3d-1h, indicates the establishment of a sulfidogenic process.

Ammonia determines transcriptional profile of microorganisms in anaerobic digestion

ABSTRACT Anaerobic digestion is important for the management of livestock manure with high ammonia level. Although ammonia effects on anaerobic digestion have been comprehensively studied, the molecular mechanism underlying ammonia inhibition still remains elusive. In this study, based on metatranscriptomic analysis, the transcriptional profile of microbial community in anaerobic digestion under low (1500 mg L-1) and high NH4 + (5000 mg L-1) concentrations, respectively, were revealed. The results showed that high NH4 + concentrations significantly inhibited methane production but facilitated the accumulations of volatile fatty acids. The expression of methanogenic pathway was significantly inhibited by high NH4 + concentration but most of the other pathways were not significantly affected. Furthermore, the expressions of methanogenic genes which encode acetyl-CoA decarbonylase and methyl-coenzyme M reductase were significantly inhibited by high NH4 + concentration. The inhibition of the co-expressions of the genes which encode acetyl-CoA decarbonylase was observed. Some genes involved in the pathways of aminoacyl-tRNA biosynthesis and ribosome were highly expressed under high NH4 + concentration. Consequently, the ammonia inhibition on anaerobic digestion mainly focused on methanogenic process by suppressing the expressions of genes which encode acetyl-CoA decarbonylase and methyl-coenzyme M reductase. This study improved the accuracy and depth of understanding ammonia inhibition on anaerobic digestion.

Methane production from four forages at three maturity stages in a ruminal in vitro system / Producción de metano de cuatro forrajes en tres estados de madurez en un sistema ruminal in vitro / Produção de metano de quatro forragens em três estágios de maturidade em um sistema ruminal in vitro

Abstract Background: Forage characteristics can modify in vitro methane production. There is little information about in vitro methane production of legumes and grasses at different maturity stages in tropical highland grazing systems. Objective: To evaluate the effect of species and forage maturity on in vitro methane production. Methods: Four forage species grown in tropical highlands of Colombia, two grasses: Kikuyu (Cenchrus clandestinus, previously named Pennisetum clandestinum) and ryegrass (Lolium perenne var. Samsum), and two legumes: Lotus (Lotus uliginosus var. Maku) and red clover (Trifolium pratense) were harvested in two paddocks at three maturity stages (young, intermediate, and mature). In vitro 48 h gas production was measured and methane proportion in gas was quantified by gas chromatography. Data were analysed as a randomized complete block (paddocks) design with a factorial arrangement 4×3 (4 species × 3 maturity stages) using the GLM procedure of SAS®. Results: Lotus produced less methane (p<0.01) than ryegrass, clover, and kikuyu (35.5 vs 64.7, 55.7 or 51.4 mL/g degraded organic matter, respectively). Younger forages produced less methane than intermediate and mature forages (42.8 vs 56.3 and 56.4 mL/g degraded organic matter, respectively). Cellulose concentration and organic matter degradability explained 67% (p<0.01) of methane production. Conclusion: Forage composition, presence of condensed tannins, and changes in fermentation patterns may explain the differences observed in in vitro methane production among species and maturity stages.

Resumen Antecedentes: Las características de los forrajes pueden afectar la producción de metano. Hay poca información sobre la producción de metano de leguminosas y gramíneas en diferentes estados de madurez en los sistemas pastoriles de trópico alto. Objetivo: Evaluar el efecto de la especie y la madurez de los forrajes sobre la producción de metano in vitro. Métodos: Cuatro especies forrajeras de trópico alto colombiano, dos gramíneas: Kikuyo (Cenchrus clandestinus, anteriormente llamado Pennisetum clandestinum) y ryegrass (Lolium perenne var. Samsum) y dos leguminosas: Lotus (Lotus uliginosus var. Maku) y trébol rojo (Trifolium pratense), fueron cosechadas de dos parcelas en tres estados de madurez (joven, intermedio y maduro). Se midió la producción de gas in vitro a las 48 h y la proporción de metano en el gas por cromatografía de gases. Los datos se analizaron empleando un diseño de bloques completos al azar (parcelas) con un arreglo factorial 4×3 (4 especies × 3 estados de madurez) mediante el procedimiento GLM de SAS®. Resultados: El lotus produjo menos metano (p<0,01) que el ryegrass, trébol o kikuyo (35,5 vs 64,7, 55,7 o 51,4 mL/g materia orgánica degradada, respectivamente). Los forrajes jóvenes produjeron menos metano que aquellos de edad intermedia y madura (42,8 vs 56,3 y 56,4 mL/g materia orgánica degradada, respectivamente). La concentración de celulosa y la digestibilidad de la materia orgánica explicaron el 67% (p<0,01) de la producción de metano. Conclusión: La composición de la pastura, la concentración de taninos condensados y los cambios en los patrones de fermentación pueden explicar las diferencias en la producción de metano in vitro según la especie y madurez del forraje.

Resumo Antecedentes: As caraterísticas das forragens podem modificar a produção de metano. Existe pouca informação sobre a produção de metano de leguminosas e gramíneas em diferentes fases de maturidade nos sistemas de pastoreio de trópico alto. Objetivo: Avaliar o efeito da espécie e a maturidade das forragens sobre a produção de metano in vitro. Métodos: Quatro espécies de forragens de trópico alto colombiano, duas gramíneas: Capim quicuio (Cenchrus clandestinus), antigamente chamado Pennisetum clandestinum) e azevém (Lolium perenne var. Samsum) e duas leguminosas: Trevina (Lotus uliginosus var. Maku) y trevo (Trifolium pratense), foram colhidas de duas parcelas diferentes em três fases diferentes de maturidade (jovem, intermedia e madura). Foi avaliada a produção de gás in vitro às 48 h e a proporção de metano foi determinada por cromatografia de gases. Os dados foram analisados com um delineamento em blocos ao acaso (parcelas) com um arranjo fatorial 4×3 (4 espécies × 3 fases de maturidade) utilizando o procedimento GLM de SAS®. Resultados: Trevina produz menos metano (p<0,01) que azevém, trevo o capim quicuio (35,5 vs 64,7, 55,7 ou 51,4 mL/g matéria orgânica degradada, respetivamente). Forragens jovens produzem menos metano que forragens com idade intermedia e matura (42,8 vs 56,3 y 56,4 mL/g matéria orgânica degradada, respetivamente). A concentração de celulosa e a digestibilidade da matéria orgânica explicaram o 67% (p<0,01) da produção de metano. Conclusão: A composição da pastagem, a concentração de taninos condensados e as mudanças nos padrões de fermentação podem explicar as diferencias na produção de metano in vitro entre espécies e maturidade de forragens.

Relationship between root exudation of organic carbon and physiological variables of irrigated rice cultivars / Relação entre exsudação radicular de carbono orgânico e variáveis fisiológicas de cultivares de arroz irrigado

ABSTRACT: Rhizospheric carbon resulting from root exudation is one of the substrates used by the soil microbiota, and reflects methane (CH4) emissions in anoxic environments such as irrigated rice cultivation. With the increase of the photosynthetic capacity of the plant in the reproductive period, there is greater accumulation of biomass which, in turn, increases the rate of root exudation. However, genotypic variations in the physiological aspects of rice plants may be related to the amount of root exudates. Ten cultivars of irrigated rice were evaluated for the exudation rate of total organic carbon (EXRToc), shoot dry matter (SDM), and physiological variables related to photosynthesis during the full flowering (blooming) period. Two experiments were conducted in the greenhouse of the Department of Soils of the UFSM (University of Santa Maria, Santa Maria, RS, Brazil) in a completely randomized experimental design. The cultivars presented significant differences in EXRToc, SDM, and all physiological variables as well as positive and significant correlations between EXRToc and physiological variables. Early cultivars were more inefficient in the physiological variables reflecting reduced values of EXRToc and SDM whereas medium-cycle cultivars were more efficient with larger EXRToc and SDM reflections.

RESUMO: O carbono rizosférico resultado da exsudação radicular é um dos substratos utilizados pela microbiota do solo, refletindo-se nas emissões de metano (CH4) em ambientes anóxicos, como no cultivo de arroz irrigado. Com o aumento da capacidade fotossintética da planta no período reprodutivo, ocorre acúmulo maior de biomassa, que por sua vez, impulsiona aumento na taxa de exsudação radicular. Entretanto, variações genotípicas nos aspectos fisiológicos das plantas de arroz podem se relacionar com a quantidade de exsudatos radiculares. Foram avaliadas dez cultivares de arroz irrigado quanto a taxa de exsudação de carbono orgânico total (TEXCOT), matéria seca da parte aérea (MSPA) e variáveis fisiológicas relacionadas com a fotossíntese no período de florescimento pleno. Dois experimentos foram conduzidos na casa de vegetação do Departamento de Solos da UFSM em delineamento experimental inteiramente casualizado. As cultivares apresentaram diferenças significativas na TEXCOT, na MSPA e em todas as variáveis fisiológicas, bem como, correlações positivas e significantes entre TEXCOT e as variáveis fisiológicas, sendo que as cultivares de ciclo precoce se mostraram mais ineficientes nas variáveis fisiológicas refletindo em valores reduzidos de TEXCOT e MSPA, enquanto que as cultivares de ciclo médio se mostraram mais eficientes com reflexos em maiores TEXCOT e MSPA.

Effects of volatile fatty acids in biohydrogen effluent on biohythane production from palm oil mill effluent under thermophilic condition

Background: Biohydrogen effluent contains a high concentration of volatile fatty acid (VFA) mainly as butyric, acetic, lactic and propionic acids. The presence of various VFAs (mixture VFAs) and their cooperative effects on two-stage biohythane production need to be further studied. The effect of VFA concentrations in biohydrogen effluent of palm oil mill effluent (POME) on methane yield in methane stage of biohythane production was investigated. Results: The methane yield obtained in low VFA loading (0.9 and 1.8 g/L) was 15­20% times greater than that of high VFA loading (3.6 and 4.7 g/L). Butyric acid at high concentrations (8 g/L) has the individual significantly negative effect the methane production process (P b 0.05). Lactic, acetic and butyric acid mixed with propionic acid at a concentration higher than 0.5 g/L has an interaction significantly negative effect on the methanogenesis process (P b 0.05). Inhibition condition had a negative effect on both bacteria and archaea with inhibited on Geobacillus sp., Thermoanaerobacterium thermosaccharolyticum, Methanoculleus thermophilus and Methanothermobacter delfuvii resulting in low methane yield. Conclusion: Preventing the high concentration of butyric acid, and propionic acid in the hydrogenic effluent could enhance methane production in two-stage anaerobic digestion for biohythane production.

Influência da razão DQO/[SO 4 2- ] e da concentração de Na + na remoção de matéria orgânica e sulfato em reator UASB / Influence of COD/[SO4 2-] ratio and Na+ concentration on organic matter and sulfate removal in UASB reactor

RESUMO O tratamento anaeróbio de efluentes industriais ricos em sulfato e sódio pode ser limitado por vários fatores, dentre os quais a relação DQO/sulfato, a concentração de SO4 2- e a concentração de sódio. Entre os fenômenos investigados estão a competição entre bactérias redutoras de sulfato e arquéias metanogênicas por substrato, a concentração de sulfeto gerado na sulfetogênese e a inibição por cátions. Este trabalho apresentou e discutiu os resultados da operação de um reator UASB, com volume útil de 10,5 L e vazão de 16,0 L.dia-1. Utilizou-se glicose, acetato e metanol (DQO≈2.000 mg.L-1), sob razão DQO/sulfato variando de 0,20 a 6,15, concentração de sulfato de 0,3 a 10,0 g.L-1 e concentração do cátion Na+ de 0,70 g.L-1 a 5,40 g.L-1. A eficiência de remoção de DQO foi mantida acima de 80%, e a concentração de sulfato removida ficou limitada a cerca de 800 mg.L-1 durante o período experimental. A atividade metanogênica específica do lodo (AME) foi de 0,630 gCH4-DQO.gSSV-1 até carga de sulfato de 0,300 gSO4 2-.L-1 e (razão DQO/sulfato de 6,15), declinando até atingir 0,168 gCH4-DQO.gSSV-1 (queda de 70%) na última fase com 10,000 gSO4 2-.L-1 (razão DQO/[SO4 2-] de 0,24) e concentração de Na+ de 5,41±0,10 g.L-1. O aumento crescente das concentrações de sódio e sulfato, mantendo-se a concentração de matéria orgânica constante, permitiu constatar a inibição da metanogênese por sódio e sulfeto, mesmo em concentrações abaixo das consideradas inibidoras individualmente. O efeito do sódio na estrutura do grânulo foi determinante na inibição da metanogênese observada.

ABSTRACT The anaerobic treatment of industrial wastewater presenting high concentrations of sulfate and sodium may be limited by several factors, including the ratio COD / [SO4 2-], the concentration of SO4 2- and the concentration of Na+. The competition between sulfate reducing bacteria and methanogenic archaea for substrate, the concentration of sulfide generated in sulfidogenesis, and the inhibition by cations are among the main phenomena that have been investigated. This paper presented and discussed the results of a UASB reactor with a volume of 10.5 L and flowrate of 16.0 L.day-1, subjected to increasing COD/sulfate ratio whereas influent COD was kept constant. Glucose, acetate and methanol (≈2,000 COD mg.L-1) were the carbon sources, and the COD/sulfate ratio ranged from 0.20 to 6.15 for sulfate concentrations of 0.3 to 10.0 g.L-1, Na + concentrations of 0.70 g.L-1 to 5.40 g.L-1. The COD removal efficiency was maintained above 80%, and the sulfate removal was limited to 800 mg.L-1 throughout the experimental period. The specific methanogenic activity (SMA) was 0.630 gCH4-DQO.gSSV-1 for 0.300 gSO4 2-.L-1 (COD/sulfate of 6.15), decreasing to 0.168 (70% decrease) in the last phase (COD /sulfate of 0.24) at concentrations of 10.000 gSO4 2-.L-1 and of Na+ 5.41±0.10 g.L-1. The progressive increase of sulfate and sodium concentrations at constant influent COD resulted in methanogenesis inhibition by sodium and sulfide, even at concentrations bellow the inhibition limits, if individually considered. The effect of sodium in the granule structure was determinant for methanogenesis inhibition.

Bacterias anaerobias: procesos que realizan y contribuyen a la sostenibilidad de la vida en el planeta / Anaerobic bacteria: processes they perform and their contribution to life sustainability on the planet

Objetivo. Se realiza una revisión sobre los aspectos degradativos de materia orgánica, obtención de energía y nutrientes de las bacterias anaerobias. La importancia de estos microorganismos es el papel que desempeñan en los procesos que contribuyen al mantenimiento de la vida misma. Dentro del metabolismo para la descomposición de macromoléculas, estos microorganismos realizan varios procesos: hidrólisis, acetogénesis y metanogénesis, entre otros, cobija reacciones que se realizan dependiendo de las características particulares de la bacteria y de las funciones que cumplen dentro del ciclo degradativo, para la obtención de nuevos productos dependiendo de las rutas bioquímicas o procesos fermentativos que allí se desarrollan.

Objective. A review of the degradation aspects of organic matter, obtaining energy and nutrients from anaerobic bacteria is performed. The importance of these organisms is the role they play in the processes contributing to the maintenance of life itself. Within the metabolism for the decomposition of macromolecules, these microorganisms perform several processes: hydrolysis acetogenesis and methanogenesis, among others, it covers reactions performed depending on the particular characteristics of the bacteria and of the roles in the degradative cycle for development of new products depending on the biochemical pathways or fermentation processes that take place there.

Inhibitory Effects of Phenolic Monomers on Methanogenesis in Anaerobic Digestion.

Aims: The purpose of this study was to evaluate the inhibitory effects of phenolic monomers on methanogenesis in anaerobic digestion and to assess the effect of hydroxyl groups’ number of phenolic monomers (aromatic structure) on inhibition of methane production by acetoclastic methanogens (archaea). Study Design: Anaerobic digestion of pig manure, anaerobic toxicity essay, The effect of the hydroxyl group’s number on the methanogenic toxicity as exhibited by monomeric tannins, Correlation of the methanogenic toxicity (IC50) with aromatic compounds hydrophobicity (logPoct), Correlation of the methanogenic toxicity (IC50) with Cresols boiling point (bp). Place and Duration of Study: Department of Chemistry, University of Kinshasa (DR Congo), between September 2011 and May 2012. Methodology: The toxicity to acetoclastic methanogenic bacteria was performed with the standard method of serum bottles; digested pig manure was utilized as inoculums and acetate as substrate. The methane gas volume produced was measured by serum bottles liquid displacement systems (Mariotte flask system). Results: The results of this study indicate that an increase in the number of hydroxyl groups on the aromatic compound was associated with a decrease in the compound’s toxicity to methanogens (archea). The toxicity of various phenolic monomers are decreasing in the following order: pyrogallol < hydroquinone < resorcinol < phenol < benzene with 3172, 2745, 1725, 1249 and 209 mg/l IC50 values respectively. A significant negative linear correlation between the toxicity of phenolic monomers together with the reference compound (benzene) and their hydrophobicity was found. Moreover, a high positive linear correlation has been found between the IC50 values of phenolic monomers and their boiling temperatures. Conclusion: The obtained results indicate that relationships exist between the phenolic monomers structure and their inhibitory effects in methane biosynthesis. The analysis of experimental results suggests that an increase in the number of hydroxyl groups on the aromatic compounds was associated with a decrease in the phenolic monomers toxicity.

Efecto del oxígeno y la glucosa en la metanogénesis y mineralización de resinas poliméricas / The effect of oxygen and glucose on polymeric resin methanogenesis and mineralisation

The effect of dissolved oxygen concentration and glucose on polymeric resin compounds’ methanogenesisand mineralisation was examined in batch cultures. They were inoculated with sludge from an upflow anaerobicsludge blanket reactor fed with polymeric resin compounds and 1.0 mg L-1 steady-state dissolved oxygen.All tests were carried out with 1,500 mg L-1 chemical oxygen demand (COD), at 30±2ºC, with 23.8 g L-1 volatile suspended solids (VSS) as inoculum and 1:1 COD/VSS ratio. The effect of different dissolved oxygen concentrations showed that COD efficiently removed glucose whilst methanogenic activity remained constant at low concentrations (0.6 and 1.0 mg L-1), but polymeric resin compounds’ COD removal efficiency increased 58.1±1% whilst methane yield decreased, due to the higher aerobic mineralisation of carbon to carbon dioxide. The result of different glucose/polymeric resin compound ratios in the presence of 0.6 mg L-1 dissolved oxygen showed that glucose did not improve polymeric resin compound removal. However, methanogenic activity decreased by 75% with polymeric resin compounds as substrate compared to methanogenic activity with glucose as sole carbon source, suggesting that the presence of glucose promotes conditions for highertolerance to oxygen. The presence of low dissolved oxygen concentrations therefore promotes polymeric resin compounds’ methanogenesis and mineralisation.

En el presente trabajo se evaluó el efecto de la concentración de oxígeno disuelto (OD) y la presencia de laglucosa en la metanogénesis y mineralización de los compuestos de resinas poliméricas, mediante cultivos en lote. El inóculo utilizado para los cultivos fueron lodos procedentes de un reactor de lecho de lodos con flujoascendente UASB (por sus siglas en inglés), alimentado con glucosa y aguas residuales de resinas poliméricas,y 1,0 mg L-1 d-1 de oxígeno disuelto en estado estacionario. Todas las pruebas fueron realizadas con 1500 mgL-1 de demanda química de oxígeno, a 30±2° C, con 23,8 g L-1 de sólidos suspendidos volátiles como inóculoy una relación demanda química de oxígeno / sólidos suspendidos volátiles de 1. El efecto de diferentesconcentraciones de oxígeno disuelto mostró que, para la glucosa, con bajas concentraciones (0,6 y 1,0 mgL-1) la eficiencia de remoción de la demanda química de oxígeno y la actividad metanogénicas permanecieronconstantes, pero la eficiencia de remoción de la demanda química de oxígeno para los compuestos de resinaspoliméricas aumentó 58,1±1% y el rendimiento de metano disminuyó, debido a que una mayor fracción delcarbono alimentado fue mineralizada a dióxido de carbono, posiblemente por una ruta aerobia. Los resultadoscon diferentes proporciones de glucosa / compuestos de resinas poliméricas (CRP) en presencia de 0,6 mg L-1 de oxígeno disuelto mostraron que la presencia de glucosa no mejoró la eliminación de los compuestosde resinas poliméricas. Sin embargo, la actividad metanogénica disminuyó en un 75% cuando solamente se usó a los compuestos de resinas poliméricas como sustrato, al compararla con la actividad metanogénica con glucosa como única fuente de carbono, sugiriendo que la glucosa no favorece la eliminación de los compuestosde resinas poliméricas, pero proporciona una mejor tolerancia al oxígeno..

Seasonal dynamics of methane in the water column of two subtropical lakes differing in trophic status / Dinâmica sazonal do metano na coluna d'água de dois lagos subtropicais em diferentes estados tróficos

Alterations in methane concentration in the water column of aquatic systems is closely linked to the processes of production and consumption of this gas, i.e., methanogenesis and methanotrophy respectively. The aim of this research is to evaluate methane dynamics through diurnal variation in the concentration of this gas in the water column of two lakes differing in trophic status at the campus of Fundação Universidade do Rio Grande (FURG). In two sampling periods (November 2001 and July 2002) methane concentrations in the water column were significantly higher (3.66 and 0.41 μmoles.L-1, respectively) at Lago dos Biguás, with mesoeutrophic features when related to Lago Polegar (1.43 and 0.19 μmoles.L-1, respectively) which has oligotrophic features. The higher methane concentrations were detected in November 2001 when higher temperature was also detected. The results highlighted the importance of trophic status as well as seasonality for the methane dynamics in these ecosystems.

As alterações na concentração de metano na coluna d'água nos ecossistemas aquáticos estão intimamente ligadas aos processos de produção e consumo deste gás, ou seja, à metanogênese e metanotrofia, respectivamente. Esta pesquisa tem como objetivo determinar a dinâmica do metano através da variação diurna da concentração deste gás na coluna d'água em dois lagos de diferentes estados tróficos do Campus da Fundação da Universidade do Rio Grande (FURG). Nos dois períodos de coleta (novembro de 2001 e julho de 2002), as concentrações de metano foram significativamente maiores no lago dos Biguás (3,66 e 0,41 μmoles.L-1, respectivamente) de caráter mesoeutrófico em relação ao lago Polegar (1,43 e 0,19 μmoles.L-1, respectivamente), de característica oligotrófica. Os maiores valores da concentração de metano na coluna d'água nos dois lagos foram observados no período de novembro de 2001, quando foram registrados também os maiores valores de temperatura. Estes resultados evidenciam tanto a importância do caráter trófico dos lagos quanto a sazonalidade em relação à dinâmica do metano nestes ecossistemas.

Variação diária da emissão de metano em solo cultivado com arroz irrigado no Sul do Brasil / Diurnal variation of methane emission from a paddy field under rice cultivation in the Southern Brazil

O metano (CH4 ) apresenta um potencial de aquecimento 23 a 32 vezes maior do que o dióxido de C e a sua emissão diária em lavouras de arroz varia com a temperatura do solo e da água, e o metabolismo das plantas. O objetivo deste estudo foi determinar o horário apropriado à coleta de amostras de ar para quantificar a emissão média diária de CH4, informação fundamental para estudos visando a derivação de índices regionais de emissão desse gás. Três campanhas de monitoramento da emissão de CH4, com coletas em intervalos de 3h, foram realizadas nos estágios de emissão da panícula (campanha 1) e de maturação de grãos (campanhas 2 e 3) do arroz sob diferentes sistemas de cultivo (convencional e plantio direto) na safra 2002/2003, na Estação Experimental do Instituto Rio Grandense do Arroz, em Cachoeirinha, RS. A coleta das amostras de ar foi realizada com seringas de poliestireno em câmaras de alumínio de 1m², sendo que as análises de metano foram realizadas por cromatografia gasosa. A emissão diária de CH4 apresentou padrão similar nas três campanhas de amostragem independente do sistema de cultivo. A faixa de emissão mínima (4-19mg CH4m-2 h-1) ocorreu entre as 24 horas e o início da manhã seguinte (9 horas), e a faixa de emissão máxima (8-33mg CH4 m-2 h-1) do início à metade da tarde (12-15 horas). A emissão de CH4 foi relacionada à temperatura da água de inundação (campanhas 1 e 3) e do solo (campanha 2). Com base nas curvas de emissão diária e considerando aspectos operacionais como a viabilidade de análise cromatográfica das amostras no período de 24h após a coleta, o intervalo entre 9 e 12 horas é recomendado para a avaliação das emissões de CH4 em lavouras de arroz irrigado quando objetiva-se a obtenção de valores equivalentes à emissão média diária desse gás. Acredita-se que a alta relação entre a temperatura e a emissão de metano valida esse procedimento para as sub-regiões produtoras de arroz no Sul do Brasil (região litorânea e Sul...

Methane (CH4) has a global warming potential 23 to 32 times higher than carbon dioxide and its emission rate in rice fields should vary daily with water and soil temperature, and plant metabolism. This study aimed to identify the appropriate time interval for air sampling in order to quantify the mean daily CH4 emission rate, key information to future studies aiming the derivation of regional indexes of CH4 emission. Three campaigns were performed to evaluate diurnal variation in CH4 emissions (3h interval) from continuously flooded rice (Oryza sativa L. cv. 'IRGA-422 CL') fields at different crop stages (panicle differentiation and ripening) and management systems (conventional tillage-CT and no-tillage-NT) during 2002/2003 crop season in long-term experiment at the Instituto Rio Grandense do Arroz (IRGA), in Cachoeirinha, RS, Brazil. Static chamber method was used and the air samples collected with polystyrene syringes were analyzed by gas chromatography. Methane emission showed a same diurnal pattern in the three campaigns in the both tillage systems. The maximum range of emission (8-33mg CH4m-2 h-1) was observed in the early afternoon (12.00 to 15.00p.m.) followed by a decline to a minimum around midnight to next morning (4-19mg CH4 m-2 h-1), in which fluxes were related to flood water temperature (campaign 1 and 3) or soil temperature (campaign 2). Taking into account the daily emission curves, plus operational aspects like chromatographic analysis of samples into the 24h period after air sampling, the time interval from 9.00 to 12.00a.m. is recommended to studies aiming to evaluate mean daily CH4 emission in rice fields. The close relationship between temperature and methane emission allows us infer about the suitability of the gas sampling procedure for the different rice sub-regions of South of Brazil (littoral region and south of RS, and south of SC) that present the same daily behavior of solar radiation and air temperature...

A Study on Cultivation of Acid-tolerant Methanogenic Granular Sludge under Low pH Values and its Characteristics / 微生物学通报

Two lab-scale expanded granular sludge bed reactors were operated using granular sludge and river sediments as seed sludge respectively, through gradually lowering down the inner pH values, acid-tolerant methanogenic granular sludge with good methanogenic activity were acquired and formed. Two EGSB reactors could be operated steadily under the condition of pH 5.8～6.2, and their volumetric loading rates were about 5.5～7.5 kg COD/(m3/d), the COD removal efficiencies were about 90%. The granular sludges taken from two reactors could maintain relatively higher methanogenic activity under low pH, and the relative activities of the granular could be 51.78% and 55.6% of the value with the condition of pH 7.0 when pH was 5.5. Studies on the diameter distribution, settling velocities, concentrations of different metal elements and microbiological characteristics of these acid-tolerant granular sludges were also conducted.

An approach to improve methanogenesis through the use of mixed cultures isolated from biogas digester.

Biogas production has been shown to be inhibited by branched chain fatty acids (isobutyric, isovaleric) produced in the digester by cellulolytic organisms. Performance of these mixed cellulolytic cultures isolated at 25°C (C25) and at 35° (C35) in a batch digester using cattle manure confirmed that C35, which forms mainly straight chain fatty acids from cellulose was more suitable for use as an inoculum than C 25 which formed predominantly branched chain fatty acids. Reconstitution of cellulolytic culture C35 and mixed methanogens M35 almost doubled both the amount and rate of methane production. Cellulolytic culture was useful in pretreatment of water hyacinth prior to its use as a substrate for methane generation. A method for preservation and transportation of mixed cellulolytic culture for use as an bineodc. ulum in the digester is described.

Inhibition of methanogenesis and its reversal during biogas formation from cattle manure.

The composition of volatile fatty acids in the biogas digester based on cattle manure as substrate and stabilised at 25°C showed that it contained 87-88% branched chain fatty acids, comprising of isobutyric and isovaleric acids, in comparison to 38 % observed in the digester operating at 35°C. Mixed cellulolytic cultures equilibrated at 25°C (C-25) and 35°C (C-35) showed similar properties, but rates of hydrolysis were three times higher than that observed in a standard biogas digester. The proportion of isobutyric and isovaleric were drastically reduced when C- 25 was grown with glucose or filter paper as substrates. The volatile fatty acids recovered from C-25 (at 25°C) inhibited growth of methanogens on acetate, whereas that from C-35 was not inhibitory. The inhibitory effects were due to the branched chain fatty acids and were observed with isobutyric acid at concentrations as low as 50 ppm. Addition of another micro-organism Rhodotorula selected for growth on isobutyric completely reversed this inhibition. Results indicate that the aceticlastic methanogens are very sensitive to inhibition by branched chain fatty acids and reduction in methane formation in biogas digester at lower temperature may be due to this effect.