Effects of cultivation conditions on the diversity of microbes involved in the conversion of rice straw to fodder

To confirm the optimum cultivation conditions for analyzing lactic acid bacterial communities and to provide the cultivation foundation for lactic acid bacterial communities that were used to convert straw into fodder, fermented rice straw was inoculated into 13 different broths. After 48 h of cultivation, pH values, volatile products, and microbial diversity were analyzed. Except for LAB broth, the pH values of the other broths could decrease to approximately 4.5. GC/MS analysis showed that lactic acid in Tomato MRS broth, MRS broth, LAB broth, and Tomato juice broth was higher than that in the other broths. DNA concentration analysis showed that the counts of microbes in Tomato MRS broth were 2.5 times of those in other broths and that tomato juice favored the reproduction of the microbes. Denaturing gradient gel electrophoresis （DGGE） analysis showed that the number of lactic acid bacterial species in HYA broth, Tomato juice broth, and Tomato MRS broth were higher than those in the other broths.     

Process of rice straw degradation and dynamic trend of pH by the microbial community MC1

The process of the rice straw degradation in the fermentor with aeration at 290 ml/h was studied. The results of dissolved oxygen （DO） indicated that the optimum DO during cellulose degradation by microbial community MC1 ranged from 0.01 to 0.12 mg/L. The change model ofpH values was as follows： irrespective of the initial pH of the medium, pH values decreased rapidly to approximate 6.0 after being inoculated within 48 h when cellulose was strongly degraded, and then increased slowly to 8.0--9.0 until cellulose was degraded completely. During the degradation process, 15 kinds of organic compounds were checked out by GC-MS. Most of them were organic acids. Quantity analysis was carried out, and the maximum content compound was ethyl acetate which reached 13.56 g/L on the day 4. The cellulose degradation quantity and ratio analyses showed that less quantity （under batch fermentation conditions） and longer interval （under semi-fermentation conditions） of rice straw added to fermentation system were contributed to matching the change model of pH, and increasing the quantity and ratio of rice straw degradation during cellulose degrading process. The highest degradation ratio was observed under the condition office straw added one time every five days （under semi-fermentation conditions）.     

Degradation of 4-CP in an internal electrolysis system

The characteristic and mechanism of parachlorophenol (4-CP) degradation in an internal electrolysis system were investigated. The degradation rate of 4-CP was higher in acid solution than that of in neutral or alkaline solution. Addition of activated carbon could make 4-CP easier be degraded by the surface contact catalysis. The dissolved oxygen in solution could take part in the electrode reaction and intensify the degradation of 4-CP. By the analysis of intermediates of degradation of 4-CP, it could be conferred that 4-CP was broken through the bond beside hydroxy firstly, then the bond beside chloride was broken and the chloride was dechlorinated simultaneously. Most intermediate products were glycerine, ethane diacid and acetic acid, while very few 1, 4-butanedial and alcohols were found.     

Characterization of phenanthrene degradation by strain Polyporus sp. S133

Polyporus sp. S133, a fungus collected from contaminated soil, was used to degrade phenanthrene, a polycyclic aromatic hydrocarbon, in a mineral salt broth liquid culture. A maximal degradation rate (92%) was obtained when Polyporus sp. S133 was cultured for 30 days with agitation at 120 r/min, as compared to 44% degradation in non-agitated cultures. Furthermore, the degradation was a ected by the addition of surfactants. Tween 80 was the most suitable surfactant for the degradation of phenanthrene by Polyporus sp. S133. The degradation rate increased as the amount of Tween 80 added increased. The rate in agitated cultures was about 2 times that in non-agitated cultures. The mechanism of degradation was determined through the identification of metabolites; 9,10-phenanthrenequinone, 2,2’-diphenic acid, phthalic acid, and protocatechuic acid. Several enzymes (manganese peroxidase, lignin peroxidase, laccase, 1,2-dioxygenase and 2,3-dioxygenase) produced by Polyporus sp. S133 were detected during the incubation. The highest level of activity was shown by 1,2-dioxygenase (187.4 U/L) after 20 days of culture.     

Enrichment of thermophilic and mesophilic microbial consortia for efficient degradation of corn stalk

Six different environmental samples were applied to enrich microbial consortia for efficient degradation of corn stalk,under the thermophilic and mesophilic conditions.The consortium obtained from anaerobic digested sludge under thermophilic condition(TC-Y)had the highest lignocellulose-degrading activity.The CO_2yield was 246.73 m L/g VS in23 days,meanwhile,the maximum CO_2production rate was 15.48 mL/(CO_2·d),which was28.75%and 52.27%higher than that under mesophilic condition,respectively.The peak value of cellulase activity reached 0.105 U/mL,which was at least 34.61%higher than the other groups.In addition,49.5%of corn stalk was degraded in 20 days,moreover,the degradation ratio of cellulose,hemicellulose and lignin can reach 52.76%,62.45%and42.23%,respectively.Microbial consortium structure analysis indicated that the TC-Y contained the phylum of Gemmatimonadetes,Acidobacteria,Chloroflexi,Planctomycetes,Firmicutes,and Proteobacteria.Furthermore,the Pseudoxanthomonas belonging to GammaProteobacteria might be the key bacterial group for the lignocellulose degradation.These results indicated the capability of degrading un-pretreated corn stalk and the potential for further investigation and application of TC-Y.     

Bioaugmentation with a pyridine-degrading bacterium in a membrane bioreactor treating pharmaceutical wastewater

The bacterial strain Paracoccus denitrificans W12, which could utilize pyridine as its sole source of carbon and nitrogen, was added into a membrane bioreactor （MBR） to enhance the treatment of a pharmaceutical wastewater. The treatment efliciencies investigated showed that the removal of chemical oxygen demand, total nitrogen, and total phosphorus were similar between bioaugmented and non-bioaugmented MBRs, however, significant removal of pyridine was obtained in the bioaugmented reactor. When the hydraulic retention time was 60 hr and the influent concentration of pyridine was 250-500 mg/L, the mean effluent concentration of pyridine without adding W12 was 57.2 mg/L, while the pyridine was degraded to an average of 10.2 mg/L with addition of W12. The bacterial community structure of activated sludge during the bioaugmented treatment was analyzed using polymerase chain reaction-denaturing gradient gel electrophoresis （PCR-DGGE）. The results showed that the W12 inoculum reversed the decline of microbial community diversity, however, the similarity between bacterial community structure of the original sludge and that of the sludge after bioaugmentation decreased steadily during the wastewater treatment. Sequencing of the DNA recovered from DGGE gel indicated that sp., Sphingobium sp., Comamonas sp., and Hyphomicrobium sp. were the dominant organisms in time sequence in the bacterial community in the bioaugmented MBR. This implied that the bioaugmentation was affected by the adjustment of whole bacterial community structure in the inhospitable environment, rather than being due solely to the degradation performance of the bacterium added.     

The contribution of mediated oxidation mechanisms in the electrolytic degradation of cyanuric acid using diamond anodes

In this work, the contribution of mediated oxidation mechanisms in the electrolytic degradation of cyanuric acid using boron-doped diamond(BDD) anodes was investigated in different electrolytes. A complete mineralization of cyanuric acid was obtained in Na Cl;however lower degrees of mineralization of 70% and 40% were obtained in Na2SO4 and Na Cl O4, respectively. This can be explained by the nature of the oxidants electrogenerated in each electrolyte. It is clear that the contribution of active chlorine(Cl2, HCl O, Cl O-)electrogenerated from oxidation of chlorides on BDD is much more important in the electrolytic degradation of cyanuric acid than the persulfate and hydroxyl radicals produced by electro-oxidation of sulfate and water on BDD anodes. This could be explained by the high affinity of active chlorine towards nitrogen compounds. No organic intermediates were detected during the electrolytic degradation of cyanuric acid in any the electrolytes, which can be explained by their immediate depletion by hydroxyl radicals produced on the BDD surface. Nitrates and ammonium were the final products of electrolytic degradation of cyanuric acid on BDD anodes in all electrolytes. In addition, small amounts of chloramines were formed in the chloride medium. Low current density(≤ 10 m A/cm2) and neutral medium(p H in the range 6–9) should be used for high efficiency electrolytic degradation and negligible formation of hazardous chlorate and perchlorate.     

Influence of lactic acid on the two-phase anaerobic digestion of kitchen wastes

To evaluate the influence of lactic acid on the methanogenesis, anaerobic digestion of kitchen wastes was firstly conducted in a two-phase anaerobic digestion process, and performance of two digesters fed with lactic acid and glucose was subsequently compared. The results showed that the lactic acid was the main fermentation products of hydrolysis-acidification stage in the two-phase anaerobic digestion process for kitchen wastes. The lactic acid concentration constituted approximately 50% of the chemical oxygen demand （COD） concentration in the hydrolysis-acidification liquid. The maximum organic loading rate was lower in the digester fed with lactic acid than that fed with glucose. Volatile fatty acids （VFAs） and COD removal were deteriorated in the methanogenic reactor fed with lactic acid compared to that fed with glucose. The specific methanogenic activity （SMA） declined to 0.343 g COD/（gVSS-d） when the COD loading were designated as 18.8 g/（L-d） in the digester fed with lactic acid. The propionic acid accumulation occurred due to the high concentration of lactic acid fed. It could be concluded that avoiding the presence of the lactic acid is necessary in the hydrolysis-acidification process for the improvement of the two-phase anaerobic digestion process of kitchen wastes.     

Products of methotrexate during chlorination

Methotrexate(MTX)is a cytotoxic drug widely used in the treatment of tumors,autoimmune diseases and severe asthma. jen00883 This drug has been frequently detected in the aquatic environment with concentrations up to μg/L levels. The MTX present in environmental water might be transformed and removed during chlorination disinfection treatment. In this work,the fate of MTX during aqueous chlorination was investigated in laboratory batch experiments,and the transformation products of MTX were identified. Aqueous solutions of MTX(1 mg/L)were chlorinated by sodium hypochlorite solution at room temperature under neutral p H conditions. Chlorinated products were pre-concentrated with solid-phase extraction(SPE)cartridges and determined by liquid chromatography electrospray ionization tandem mass spectrometry(LC–ESI–MS/MS). The reaction of MTX chlorination exhibited pseudo-first-order kinetics and the half-life time of MTX degradation was calculated to be 1.65 min,when the initial chlorine concentration was 2 mg/L. Two chlorinated MTX congeners,4-amino-3-chlorinated-N10-methylpteroylglutamic(monochloro-MTX)and 4-amino-3,5-dichloro-N10-methylpteroylglutamic(dichloro-MTX)were found in the chlorinated solution. Monochloro-MTX was successfully fractionated by high performance liquid chromatography(HPLC)and its structure was further identified using ~1H nuclear magnetic resonance(NMR)analysis. The presence of the two products in real hospital wastewater was then examined and both compounds were detected. Finally,the effects of MTX and monochloro-MTX on the cell cycle progression in vitro were evaluated using zebrafish liver cell line. It was found that both compounds could inhibit the proliferation of zebrafish liver cells through S phase arrest and their effects on the cell cycle profile had no significant difference.     

Biodegradation of organochlorine pesticide endosulfan by bacterial strain Alcaligenes faecalis JBW4

The recently discovered endosulfan-degrading bacterial strain Alcaligenesfaecalis JBW4 was isolated from activated sludge. This strain is able to use endosulfan as a carbon and energy source. The optimal conditions for the growth of strain JBW4 and for biodegradation by this strain were identified, and the metabolic products of endosulfan degradation were studied in detail. The maximum level of endosulfan biodegradation by strain JBW4 was obtained using broth at an initial pH of 7.0, an incubation temperature of 40℃ and an endosulfan concentration of I00 mg/L. The concentration of endosulfan was determined by gas chromatography. Strain JBW4 was able to degrade 87.5% of α-endosulfan and 83.9% of β-endosulfan within 5 days. These degradation rates are much higher than the previously reported bacterial strains. Endosulfan diol and endosulfan lactone were the major metabolites detected by gas chromatography-mass spectrometry; endosulfan sulfate, which is a persistent and toxic metabolite, was not detected. These results suggested that A. faecalis JBW4 degrades endosulfan via a non-oxidative pathway. The biodegradation of endosulfan by A. faecalis is reported for the first time. Additionally, the present study indicates that strain JBW4 may have potential for the biodegradation of endosulfan residues.     

纤维素分解菌复合系MC1分解木薯淀粉厂残渣

木薯渣木质纤维素含量较高,是有开发潜力的生物质资源,目前,木薯渣主要用来生产饲料和酒精.用其进行甲烷发酵也是很有前景的应用途径,但是木薯渣含水量大,颗粒分散,不利于固体发酵.利用纤维素分解细菌复合系MC1分解木薯渣,可将其转化为可溶的小分子有机物,提高其甲烷发酵效率.在分解过程中测定了木薯渣各成分的减少和分解产物.结果显示,经18 d分解,木薯渣的总干重减少了47.3%,其中纤维素减少了22.7%,半纤维素减少了90.4%,木质素减少了11.3%,其总分解量的85%在前6 d被分解.MC1分解木薯渣过程中可溶性物质总含量开始为18%,3 d后达到最高,为33%.GC-MS测定挥发性产物的总量在培养6 d后达到最高,挥发性产物共检测出12种,其中量较大的化合物为乙醇、乙酸、乙二醇、丁酸及甘油,这些产物为甲烷发酵菌可直接利用或者容易转化为甲烷菌直接利用的成分.因此利用MC1分解木薯渣作为甲烷发酵的前发酵手段将很有应用前景.     

玉米秸秆发酵浸出液模拟废水发酵产氢的放大实验研究

在20L的半连续流发酵罐中,以牛粪堆肥为产氢菌源,按照玉米秸秆发酵浸出液的主要成份配制模拟废水,考察和分析了几个关键环境因素对发酵产氢的影响。结果表明,HRT、C/N、Fe2+浓度和模拟废水浓度对发酵产氢均有不同程度的影响。在本实验条件下,秸秆模拟废水的氢产量、氢浓度和产氢速率分别为11.80mol/kg、56%和8.81L/(L?d),底物转化率大于90%,废水中COD去除率为39.40%。在整个发酵产氢过程,液相主要发酵副产物为丁酸、乙酸和丙酸以及少量的乙醇和丁醇。     

木质纤维素分解菌复合系WSC-6分解稻秆过程中的产物及pH动态

为了探明WSC-6分解稻秆过程中的代谢特性,本试验以稻杆为唯一碳源,50℃静止培养WSC-6,以一次性添加不同量稻秆和多次连续添加的方式,研究了WSC-6分解稻杆的相对分解率、绝对分解量、分解产物及pH变化特性.结果表明,一次性添加1%稻秆时,pH由初始的7.8迅速下降,第3 d降到6.0后逐渐回升,6 d后稳定在8.0左右,在此过程中DO值保持在0.01～0.12 mg·L-1微好氧条件.以GC-MS法在稻杆分解过程中检测到乙醇、乙酸、乳酸、甘油等10种以上有机物,其中乳酸的含量最高,为7.381 g·L-1.在90 d的实验中,一次性添加不同量稻秆时,随着稻秆量的增加,pH下降得快而低,且回升时间拖后,当稻秆量为5%时pH下降到5.0后不再回升.在多次连续添加稻秆的实验中,隔12d和15d添加的处理pH重复下降和回升的规律性变化,分解活性保持旺盛.90 d后隔15 d添加的处理相对分解率最高,为86.7%;绝对分解量以隔6 d添加的处理最高,为32.4 g.pH变化规律能够反映WSC-6对木质纤维素的分解进程及分解活性.     

玉米秸秆化学预处理后进行厌氧干发酵试验的研究

采用稀硫酸和稀氢氧化钠处理玉米秸秆,分析秸秆纤维组成和纤维结构的变化,并对酸、碱处理后的玉米秸秆进行厌氧干发酵试验。研究表明,采用稀酸处理玉米秸秆的还原糖产率明显高于采用稀碱处理的玉米秸秆;稀硫酸对半纤维素的去除率可达74.27%,稀碱对木质素的去除率最高可达83.94%;玉米秸秆经稀酸处理和稀碱处理后进行厌氧干发酵试验,其累积产气率和甲烷含量都有所提高,但酸法高于碱法。     

腐殖酸强化六氯苯厌氧降解规律及其中间产物

为探索厌氧条件下HA(腐殖酸)对HCB(六氯苯)降解的影响,通过构建ρ(HA)为0(HA₀)、120(HA₁₂₀)及200(HA₂₀₀)mg/L厌氧发酵体系,利用气相色谱-电子捕获检测器(GC-ECD)对ρ(HCB)及中间产物进行测定. 结果表明:发酵系统运行72 h后,HA₀、HA₁₂₀、HA₂₀₀处理中HCB降解率分别为13.0％、16.5％和17.4％,表明HA可以促进HCB的降解,并且随ρ(HA)的增加,降解效果越好,但降解效率增幅降低. 对HCB脱氯中间产物的分析发现,PeCB(五氯苯)、TeCB(四氯苯)、TCB(三氯苯)和DCB(二氯苯)均有检出;在发酵的前12 h,ρ(DCB)快速增加,发酵72 h后HA₀、HA₁₂₀、HA₂₀₀处理中ρ(DCB)分别为50.6、51.0和57.6 μg/L,明显高于其他中间产物,表明DCB是HCB脱氯的限速中间体;对DCB进行同分异构体分析表明,DCB的同分异构体有1,2-DCB、1,3-DCB和1,4-DCB三种,发酵结束后HA₀、HA₁₂₀和HA₂₀₀处理中ρ(1,4-DCB)最高,分别为22.0、17.2和18.7 μg/L,1,4-DCB为HCB降解的主要中间产物. 研究显示,通过调控厌氧发酵体系内HA的含量可达到强化HCB脱氯的目的.      

秸秆发酵乳酸菌复合系SFC-2的构建及其组成多样性研究

为了获得促进作物干秸秆乳酸发酵的微生物,以玉米秸秆和水稻秸秆的自然发酵物为菌种来源,用MRS蔗糖培养基,通过连续定向继代培养,筛选出pH下降迅速、乳酸含量高、组成稳定的乳酸菌复合系SFC-2.DGGE分析表明,SFC-2经过连续继代培养,从第25代开始其微生物组成基本稳定.SFC-2 培养12 h后pH下降至3.8,乳酸含量达10.64 mg/mL,其中64%为L-(+) 乳酸.通过平板分离获得4株细菌,全部为Lactobacillus,其近缘种分别为L. fermentum、L. plantarum、L. paracasei和L. paracasei sub sp.;通过16S rDNA克隆文库分析获得7个克隆,其近缘种主要为L. fermentum、L. plantarum及L.paracasei;在16S rDNA的克隆文库中,76.3%为L. fermentum的近缘种,20.3%为L. plantarum的近缘种,3.4%为L. paracasei的近缘种.     

高效纤维素分解菌在蔬菜-花卉秸秆联合好氧堆肥中的应用

以滇池流域典型的蔬菜废物和花卉秸秆为堆肥原料,以本实验室筛选、保存的17株纤维素降解菌和1株购买的产黄纤维单胞菌(Cellulomonas Flavigena)为复合接种剂,对不同接种条件和控温条件下的联合堆肥中试进行了研究.实验结果表明,在一次发酵的初始阶段,以体积分数0.5%的接种量向堆肥中接种纤维素降解复合菌剂可有效提高发酵过程堆料中纤维素降解菌的种群密度,并使其迅速成为优势菌群,尤其是当堆体处于控温55℃的工况条件时,其菌群密度可保持在3.84×10⁹～1.80×10¹⁰CFU/g;在二次发酵的初始阶段,以体积分数1%的接种量接种,可有效提高二次发酵阶段堆温的回升.对堆料中木质素和纤维素含量以及堆肥终产物的粒径分布指标--过筛率的检测表明,接种的复合纤维素降解菌可有效地降解堆料中的木质纤维素,接种处理中纤维素的降解率比不接种处理高23.64%,接种处理堆肥终产物的过筛率(2.0 cm)比不接种处理高18.28%.研究表明,用纤维素降解复合菌剂进行二次接种二次发酵,能够有效地促进蔬菜-花卉秸秆联合好氧堆肥中物料的纤维素组分的降解,达到加快堆肥进程,提高堆肥品质的目的.     

pH值调控柠檬酸污泥厌氧发酵产酸及碳源潜力研究

以柠檬酸废水厌氧颗粒污泥为接种物,在不同pH值调控条件下开展柠檬酸生产废水剩余活性污泥厌氧发酵产酸研究。通过对发酵液挥发性脂肪酸(VFAs)、有机质、氮磷和污泥脱水性能的分析,探讨了柠檬酸污泥厌氧产酸机制。结果表明,pH³10的碱性条件更有利于有机质的溶出从而促进VFAs的产生。三维荧光光谱分析发现在恒定pH值下腐殖酸(HA)和富里酸(FA)会大量溶出降低VFAs的产量。初始pH=10是柠檬酸污泥厌氧产酸的最佳pH值,发酵4d的VFAs浓度最高达(6681.47±126.82)mg COD/L,是文献报道中市政污泥产酸量的近2倍,其中乙酸占比49.8%,发酵后产酸功能菌Chloroflexi、Bacteroidota的相对丰度分别由初始的9.52%、10.87%增至16.84%、14.39%,污泥归一化毛细吸水时间(n_CST)为(11.34±0.27)s×L/g,脱水性能良好,发酵液TP浓度为(20.45±0.33)mg/L。研究表明,利用柠檬酸剩余活性污泥碱性厌氧发酵产酸作为污水处理过程中的外加碳源具有较大潜力。     

PLFA法研究稻草固态发酵中的微生物群落结构变化

在稻草固态发酵体系中同时接种土壤微生物和黄孢原毛平革菌,用磷脂脂肪酸（PLFA）谱图分析法研究发酵过程的微生物群落和生物量变化,同时监测木质纤维素降解率的变化.结果表明,发酵后木质纤维素的降解率可达44%.根据标记性脂肪酸的变化,在发酵第6 d,革兰氏阳性菌、革兰氏阴性菌、真菌的含量都达到了最高值,其中,革兰氏阳性菌的含量较低;真菌和细菌的脂肪酸含量比值变化范围为0 .2～0 .5,说明真菌是降解木质纤维素的主要群落.主成分分析结果显示,发酵后期以18碳不饱和脂肪酸为主,与标记性脂肪酸分析结果一致,同时跟木质纤维素降解率的变化趋势对应,因此PLFA分析法可以较好地反映稻草固态发酵过程中的微生物群落结构和生物量的变化.     

Microbial population dynamics and changes in main nutrients during the acidification process of pig manures

This study evaluated the impact of pig manure acidification on anaerobic treatment and composition of the fecal microbial community.According to the different chemical oxygen demand (COD) in the anaerobic treatment processes,pig manure was diluted 4 times (×4),16 times (×16),or 64 times (×64) and subjected to acidification.During the acidification process,pH,soluble chemical oxygen demand (SCOD),volatile fatty acids (VFAs),nitrogen (N),phosphorus (P) and potassium (K) were determined along with microbial po...