Long-term effect of inoculum pretreatment on fermentative hydrogen production by repeated batch cultivations: homoacetogenesis and methanogenesis as competitors to hydrogen production

Long-term effects of inoculum pretreatments (heat, acid, loading-shock) on hydrogen production from glucose under different temperatures (37 °C, 55 °C) and initial pH (7 and 5.5) were studied by repeated batch cultivations. Results obtained showed that it was necessary to investigate the long-term effect of inoculum pretreatment on hydrogen production since pretreatments may just temporarily inhibit the hydrogen consuming processes. After long-term cultivation, pretreated inocula did not enhance hydrogen production compared to untreated inocula under mesophilic conditions (initial pH 7 and pH 5.5) and thermophilic conditions (initial pH 7). However, pretreatment could inhibit lactate production and lead to higher hydrogen yield under thermophilic conditions at initial pH 5.5. The results further demonstrated that inoculum pretreatment could not permanently inhibit either methanogenesis or homoacetogenesis, and methanogenesis and homoacetogenesis could only be inhibited by proper control of fermentation pH and temperature. Methanogenic activity could be inhibited at pH lower than 6, both under mesophilic and thermophilic conditions, while homoacetogenic activity could only be inhibited under thermophilic condition at initial pH 5.5. Microbial community analysis showed that pretreatment did not affect the dominant bacteria. The dominant bacteria were Clostridium butyricum related organisms under mesophilic condition (initial pH 7 and 5.5), Thermoanaerobacterium sp. related organisms under thermophilic condition (initial pH 7), and Thermoanaerobacterium thermosaccharolyticum related organisms under thermophilic condition (initial pH 5.5). Results from this study clearly indicated that the long-term effects of inoculum pretreatments on hydrogen production, methanogenesis, homoacetogenesis and dominant bacteria were dependent on fermentation temperature and pH.     

The effect of concentration of phenols on their batch methanogenesis

The biodegradability of phenol and six other phenolic compounds (o-, m-, and p-cresol, 2-, 3-, and 4-ethylphenol) was examined in batch methanogenic cultures. The effect of concentration of these alkyl phenols on the anaerobic biodegradation of phenol was also evaluated. The inoculum used in this study was cultivated in a continuous flow laboratory fermenter with phenol as the primary substrate. Phenol, at initial concentrations as high to 1400 mg/L was completely degraded to methane and carbondioxide after 350 hours incubation. Complete degradation of m- and p-cresol was also observed while the ethylphenols and o-cresol were not significantly degraded.At initial concentrations exceeding 600 mg/L, phenol inhibited the phenol-degrading microorganisms but not the methanogens. At about 600 mg/L, cresols reduced the rate of phenol degradation to 50% of that observed in a control culture containing only 200 mg/L phenol. Ethylphenols were more inhibitory than cresols. Phenol degrading microorganisms were more susceptible to inhibition by cresols and ethylphenols than were the methanogens. The inhibitory effects of the three isomers of cresol and ethylphenol did not vary with the isomer but rather with the substituted functional group.     

Evaluation of pretreatment methods on mixed inoculum for both batch and continuous thermophilic biohydrogen production from cassava stillage

Anaerobic sludges, pretreated by chloroform, base, acid, heat and loading-shock, as well as untreated sludge were evaluated for their thermophilic fermentative hydrogen-producing characters from cassava stillage in both batch and continuous experiments. Results showed that the highest hydrogen production was obtained by untreated sludge and there were significant differences (p < 0.05) in hydrogen yields (varied from 32.9 to 65.3 mlH₂/gVS) among the tested pretreatment methods in batch experiments. However, the differences in hydrogen yields disappeared in continuous experiments, which indicated the pretreatment methods had only short-term effects on the hydrogen production. Further study showed that alkalinity was a crucial parameter influencing the fermentation process. When the influent was adjusted to pH 6 by NaHCO₃ instead of NaOH, the hydrogen yield increased from about 40 to 52 mlH₂/gVS in all the experiments. Therefore, pretreatment of anaerobic sludge is unnecessary for practical thermophilic fermentative hydrogen production from cassava stillage.     

The effect of phenolic acids and molasses spent wash concentration on distillery wastewater remediation by fungi

The effect of gallic acid, vanillic acid, and molasses spent wash (MSW) concentration on growth and decolourizing capability of four fungi (Geotrichum candidum, Coriolus versicolor, Phanerochaete chrysosporium and Mycelia sterilia) was studied. Fungal growth was inhibited to a varying extent in the presence of gallic and vanillic acid, except for G. candidum, which was unaffected by gallic acid. G. candidum and P. chrysosporium growth rates increased in the presence of increasing concentrations of MSW (up to 50% v/v), however, growth of M. sterilia and C. versicolor was inhibited at spent wash concentrations above 5% (v/v). Increasing the concentration of MSW from 6·25% (v/v) to 12·5% (v/v) increased the decolourizing ability of each fungus, except for M. sterilia. C. versicolor exhibited greatest colour removal with a reduction of 0·43 units at A₄₇₅ (equivalent to 53% colour reduction) after 10 days growth in 12·5% (v/v) MSW.     

Coenzymes of methanogenesis from hydrogen and carbon dioxide

Methanogenic bacteria gain their energy for growth from the conversion of a number of simple carbon compounds to methane. With a few exceptions all species known to date are able to reduce CO₂ at which hydrogen acts as the electron donor. The reduction of CO₂ can formally be considered to proceed through the formyl, the formaldehyde and the methyl level of reduction. These C₁-units do not occur as free intermediates, but they remain bound to a number of unique coenzymes during the process. In this paper a survey is given of the structures and functions of these compounds; it deals with methanopterin derivatives, carbon dioxide reduction (CDR) factor, factor F₄₃₀ and coenzyme M derivatives. A model of the process of methanogenesis that integrates previous ones and that allocates a function to the various coenzymes is presented.This paper is adapted from a treatise by the same author, entitled Coenzymes of methanogenesis. that was awarded the Kluyver prize 1984 by the Netherlands Society for Microbiology.     

A consolidated bio-processing of ethanol from cassava pulp accompanied by hydrogen production

A biphasic fermentation approach was undertaken for the production of ethanol and hydrogen from cassava pulp. The glucose generated by co-culture of Clostridium thermocellum and Thermoanaerobacterium aotearoense was 13.65±0.45 g L(-1), which was 1.75 and 1.17-fold greater than that produced by mono-cultures of C. thermocellum and T. aotearoense, respectively. The accumulated glucose could be utilised rapidly by subsequently inoculated Saccharomyces cerevisiae. An inoculum ratio of 1:1, a thermophilic fermentation of 84 h, and a pulp concentration of 4% proved optimal for ethanol production, fermentation efficiency, and productivity. With these conditions, the ethanol level reached 8.83±0.31 g L(-1) with a fermentation efficiency of 64.95±2.71%. Hydrogen production of 4.06 mmol by the co-culture system was 1.54 and 2.09-fold greater than that produced by mono-cultures of C. thermocellum and T. aotearoense, respectively. This sequential co-culture approach provided a consolidated bio-processing means to produce ethanol and hydrogen from cassava pulp.     

The effect of energy substrate concentration and amino acids on the in vitro development of preimplantation porcine embryos

As the pig becomes increasingly used for biomedical research, an effective and efficient in vitro culture system is essential. This study aimed to improve the commonly used porcine embryo culture medium, NCSU23, by altering the energy substrates and adding amino acids, using electrically activated diploid parthenotes from oocytes obtained from the ovaries of prepubertal and adult animals. Morphological development to day 6 and blastocyst cell number were examined. Glucose (5.56 mM) was replaced by pyruvate and lactate (0.2 mM and 5.7 mM, respectively) for either the entire culture period or for the first 48 h only. Blastocyst rates were not different between any of the treatments, and were similar for prepubertal and adult oocytes. When the embryos were cultured with pyruvate and lactate for the first 48 h and then glucose, there was a significant increase in blastocyst cell number compared to glucose only. Blastocysts produced using pyruvate and lactate for the entire time tended to have more cells than those exposed to glucose only and less than those who were cultured in pyruvate and lactate for the first 48 h and then glucose. Nonessential amino acids added for the first 48 h and nonessential and essential amino acids added for the remaining time significantly increased blastocyst cell number only when the embryos were grown in pyruvate and lactate followed by glucose. Blastocyst rates were not different between any of the treatments, and this result was the same when using sow or gilt oocytes. The modified medium was then tested using in vitro matured and fertilized embryos from sow oocytes. Blastocyst rates and cell number were significantly increased in the modified medium compared to those grown in unmodified NCSU23. This shows that altering energy substrates and adding amino acids can increase the quantity and cell number of IVP blastocysts compared with NCSU23.     

The effect of aromatic structure on the inhibition of acetoclastic methanogenesis in granular sludge

Summary Benzene derivatives are important constituents of certain effluents discharged by pulp and paper, petrochemical and chemical industries. The anaerobic treatment of these waste-waters can be limited due to methanogenic inhibition exerted by aromatic compounds. The objective of this study was to evaluate the effect of aromatic structure on acetoclastic methanogenic inhibition. The toxicity to acetoclastic methanogens was assayed in serum flasks utilizing granular sludge as inoculum. Among the monosubstituted benzenes, chlorobenzene, methoxybenzene and benzaldehyde were the most toxic with 50% inhibition occurring at concentrations of 3.4, 4.2 and 5.2 mm, respectively. In contrast, benzoate was the least inhibitory: concentrations up to 57.3 mm were non-toxic. In general, the toxicity of aromatic compounds increased with increasing length of aliphatic side-chains, increasing the number of alkyl or chlorine groups. The logarithm of the partition coefficient octanol/water (log P), an indicator of hydrophobicity, was observed to be positively correlated with the methanogenic inhibition. The results indicate that hydrophobicity is an important factor contributing to the high toxicity of the most inhibitory aromatic compounds.     

Effects of temperature,inoculum size and starch hydrolyzate concentration on butanediol production by Bacillus licheniformis

An optimization study has been performed on 2,3-butanediol production by Bacillus licheniformis NCIMB 8059 from different carbon sources (glucose, sucrose and cornstarch hydrolyzate), alternately varying temperature (34相似文献   

初始底物浓度对序批式培养光合细菌产氢动力学影响

实验研究了初始底物浓度对序批式培养光合细菌生长、降解及产氢过程的影响,根据最大比生长速率实验数据拟合得到其关于初始底物浓度影响的关联式,并在建立的修正Monod模型基础上建立了光合细菌比生长速率、基质比消耗速率和比产氢速率关于底物初始浓度影响的数学模型,模型预测值与实验结果在光合细菌生长期和稳定期内得到较好吻合,反映了光合细菌生长、降解和产氢过程中受底物初始浓度限制性和抑制性影响的基本规律。分析发现光合细菌生长、降解基质和产氢过程中最适底物浓度为50 mmol/L,初始底物浓度低于或高于该浓度时,光合细菌生长、降解及产氢过程都受到限制性或抑制性影响,且抑制性影响较限制性影响效果更明显;底物比消耗速率受初始底物浓度影响较小。     

Effect of clay pretreatment on photofermentative hydrogen production from olive mill wastewater

The aim of this paper was to gain further insight into the effect of the clay pretreatment process on photofermentative hydrogen production. This two-stage process involved a clay pretreatment step followed by photofermentation which was performed under anaerobic conditions with the illumination by Tungsten lamps. Rhodobacter sphaeroides O.U.001 was used for photofermentation. Higher amounts of color (65%), total phenol (81%) and chemical oxygen demand (31%) removal efficiencies were achieved after clay pretreatment process. During photofermentative hydrogen production with the effluent of clay pretreatment process, the main organic compounds resulting higher hydrogen production rates were found to be acetic, lactic, propionic, and butyric acids. Compared to photofermentation using raw olive mill wastewater ( 16LH2/LOMW), the amount of photofermentative hydrogen production was doubled by using the effluent of the clay pretreatment process (31.5LH2/LOMW). The reasons for the improvement of hydrogen production by clay treatment can be attributed to the high removal of the hardly biodegradable compounds such as phenols; minor removal of organic acids, sugars and amino acids that are known to enhance photofermentative hydrogen production; and the color depletion of raw OMW which might cause a shadowing effect on the photosynthetic bacteria.     

The effect of pH on ruminal methanogenesis

Abstract: When a fistulated cow was fed an all forage diet, ruminal pH remained more or less constant (6.7 to 6.9). The ruminal pH of a concentrate-fed cow decreased dramatically in the period soon after feeding, and the pH was as low as 5.45. Mixed ruminal bacteria from the forage-fed cow converted CO₂ and H₂ to methane, but the ruminal fluid from the concentrate-fed cow did not produce methane. When the pH of the ruminal fluid from the concentrate-fed cow was adjusted to pH 7.0, methane was eventually detected, and the absolute rate constant of methane production was as high as the one observed with ruminal fluid from the forage fed cow (0.32 h⁻¹). Based on the zero-time intercepts of methane production, it appeared that the concentrate-fed cow had fewer methanogens than the forage-fed cow. When the mixed ruminal bacteria were incubated in a basal medium containing 100 mM acetate, methanogenesis was pH-dependent, and no methane was detected at pH values less than 6.0. Because the removal of acetic acid completely reversed the inhibition of methanogenesis, it appeared that volatile fatty acids were causing the pH-dependent inhibition. Based on these results, concentrate diets that lower ruminal pH may provide a practical means of decreasing ruminal methane production.     

Medium chain carboxylic acids production from waste biomass: Current advances and perspectives

Alternative chemicals to diverse fossil-fuel-based products is urgently needed to mitigate the adverse impacts of fossil fuel depletion on human development. To this end, researchers have focused on the production of biochemical from readily available and affordable waste biomass. This is consistent with current guidelines for sustainable development and provides great advantages related to economy and environment. The search for suitable biochemical products is in progress worldwide. Therefore, this review recommends a biochemical (i.e., medium chain carboxylic acids (MCCAs)) utilizing an emerging biotechnological production platform called the chain elongation (CE) process. This work covers comprehensive introduction of the CE mechanism, functional microbes, available feedstock types and corresponding utilization strategies, major methods to enhance the performance of MCCAs production, and the challenges that need to be addressed for practical application. This work is expected to provide a thorough understanding of the CE technology, to guide and inspire researchers to solve existing problems in depth, and motivate large-scale MCCAs production.     

In vitro production of short-chain fatty acids from resistant starch by pig faecal inoculum

The need to improve the knowledge of fermentation processes within the digestive tract in pigs is growing, particularly for ingredients that may act as potential prebiotic sources, such as resistant starch (RS). A study (based on enzymatic digestion followed by in vitro fermentation) was conducted to investigate whether various sources of RS, obtained from eight native starches characterized by inherent heterogeneous starch chemistry and structure, can influence short-chain fatty acid (SCFA) concentrations and relative production kinetics. Total and individual SCFA productions were evaluated over time and up to 72 h of incubation. The in vitro hydrolysis of native starches allowed a classification from very high [⩾650 g/kg dry matter (DM)] to low (<50 g/kg DM) RS amount. The total SCFA production was similar between ingredients, whereas acetate and butyrate molar ratios in the SCFA profile differed (from 0.48 to 0.56 and from 0.17 to 0.25, respectively; P < 0.05). Differences in fermentation kinetic parameters for total and individual SCFA productions were observed (P < 0.05). Considering the total SCFA production after 72 h of incubation, the time at which half of the maximum production has been reached (T_1/2), the maximum rate of production (R_max) and its time of occurrence (T_max) differed between ingredients (P < 0.05), with values ranging from 6.1 to 11.9 h, from 0.459 to 1.300 mmol/g DM incubated per hour and from 5.1 to 9.8 h, respectively. Overall, a similar trend was observed considering individual SCFA productions. In particular, T_1/2 ranged from 6.4 to 12.5 h, from 5.5 to 12.5 h and from 6.7 to 11.3 h for acetate, propionate and butyrate, respectively (P < 0.05). For R_max, differences were obtained for propionate and butyrate productions (P < 0.05), whereas no difference was recorded for acetate. In summary, our findings indicated that both quantitative and qualitative production of SCFA and related kinetics were influenced by fermentation of RS obtained from native starches characterized by heterogeneous starch characteristics. Current findings are based on an in vitro approach, and thus require further in vivo validations.     

Deterrent effect of carboxylic acids on cabbage root fly oviposition

In laboratory experiments, 11 selected carboxylic acids were tested to determine which part of the sinapic acid molecule is responsible for deterring cabbage root fly from laying its eggs on otherwise-acceptable cauliflower host-plants. The deterrent effect was only obtained with compounds containing at least one carboxylic group in the molecule. Hence, the aliphatic acids were as deterrent as the aromatic acids to the fly and all the carboxylic acids were as deterrent as sinapic acid, reducing oviposition by > 50%. The inclusion of two carboxylic groups in the molecule, (e.g. phthalic acid and oxalic acid) did not increase the deterrent effect observed with sinapic acid. Some of the long chain fatty acids, with low volatility, low water solubility and thus greater persistence, offer practical opportunities for deterring Delia radicurn from laying its eggs on plants in the field.     

The effect of inoculum density and conditioned medium on the production of ajmalicine and catharanthine from immobilized Catharanthus roseus cells

The effect of the cell-inoculum size and the addition of conditioned medium on ajmalicine and catharanthine production were studied using immobilized Catharanthus roseus cells. Higher specific-uptake rates of ammonium, nitrate, and sugars were observed in the low-inoculum-density cultures (50 g FW/L) compared to the high-inoculum-density cultures (100 g FW/L). Alkaloid production was not correlated with the exhaustion of a particular nutrient from the medium. The high-inoculum-density cultures produced higher ajmalicine concentrations throughout the experiment. Catharanthine production was similar between the two inoculum-density cultures. The addition of conditioned medium to MS-production medium dramatically improved the production of ajmalicine and catharanthine. The addition of conditioned medium enhanced ajmalicine production from immobilized Catharanthus roseus cultures on day 15 by at least two- to fourfold compared to media without the conditioning factors. Catharanthine production was increased by nearly fivefold in cultures with conditioned medium compared to those without conditioned medium. The enhancing effects of conditioned medium on alkaloid production were attributed to an unidentified factor produced and secreted by suspension cultures of C. roseus. The presence of conditioned medium also decreased the sucrose hydrolysis rate. The ajmalicine concentration in these immobilized cell cultures was found to be a function of the fresh-weight concentration, irrespective of the inoculum density or the culture medium. The medium choice and the inoculum density determined how rapidly fresh weight was accumulated and thus, how quickly ajmalicine was produced. Ajmalicine production correlated positively with fresh-weight concentration, but catharanthine production was not correlated with fresh-weight concentration.