Phosphate removal using aerobic bacterial consortium and pure cultures isolated from activated sludge

In the present study, an attempt is made to evaluate the kinetics of biological phosphate removal using a bacterial consortium of activated sludge, as well as screening for dominant polyphosphates accumulating bacteria. The results showed an efficient phosphate uptake (P < 0.001) of the consortium, with rates related to the initial concentration of both phosphate and carbon sources. Short chain volatile fatty acids presented the suitable substrates for enhanced biological phosphorus removal, of which maximum yield reached 99.23% and 78.51% in basal salt medium supplemented with 0.5% of sodium acetate and lactate respectively. Fifteen phosphate-accumulating bacteria were isolated from the activated sludge and only four isolates were selected and characterized as Pseudomonas aeruginosa AS1, Moraxella lacunata AS2, Acinetobater junii AS3 and Alcaligenes denitrificans AS4. The highest efficiency of phosphate uptake using pure culture was achieved with Ac. junii AS3 (83.36) followed by P. aeruginosa AS1 (81.78%), Al. denitrificans AS4 (76.72%), and M. lacunata AS2 with 50.6%.     

Dynamics of bacterial community of Suillus granulatus mushroom shiro in Pine (Pinus tabuliformis) forests北大核心CSCD

To evaluate bacterial community variation in the mushroom shiro of Suillus granulatus during fruiting, we collected soil samples from the mushroom shiro in the pine (Pinus tabuliformis) forest of mountainous area in Beijing from May to November and evaluated the bacterial community using polymerase chain reaction - denaturing gradient gel electrophoresis (PCR-DGGE). Total soil DNA was extracted using a commercial soil DNA isolation kit. PCR amplification and DGGE were performed using bacterial universal primers 338F and 518R. The specific bands were excised from the gel and sequenced. The results revealed that soil bacterial community maintained considerably high level and changed seasonally with the mushroom fruiting. In total, 53 bands of DGGE profiles were sequenced and divided into 5 phyla (Acidobacteria, Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria and 22 genera (Acidobacterium, Aminobacter, et al). Species from Proteobacteria and Acidobacteria were the dominant bacterial groups sharing considerably high relative abundance, while class a-Proteobacteria was the most abundant group. The variation of the relative abundance of γ-Proteobacteria species was consistent with the mushroom fruiting season. The relative abundance of Acidobacteria species obviously increased before mushroom flush (in July). The fruiting of S. granulatus and the relative abundance of γ-Proteobacteria were correlated with each other. The present study provided a basis for conservation and domestication of mushroom S. granulatus.     

青霉素菌渣资源化为饲料原料的营养价值研究

为探讨青霉素菌渣资源化为饲料原料潜在的营养价值,本研究以抗生素制药厂发酵剩余培养基(即青霉素菌渣)为研究对象,利用元素分析仪、电感耦合等离子发射光谱对菌渣中的非金属元素、金属元素进行了测定,并利用氨基酸分析仪、凯氏定氮法、DNS法、索氏提取法对其基本营养物质氨基酸、粗蛋白、总糖及脂肪含量进行了简单分析。结果表明:C、N、S、H 4种非金属元素含量较高,其中,动物体有机体常量元素C含量最高,质量分数为34.52%。相对而言,金属元素含量较低,其中K、Na等常量元素含量较高,4种重金属污染物Cd、Cr、Hg、Pb含量与相关国家饲料安全标准对照而得,均低于下限水平。此外,以蛋白质、脂肪和总糖含量为考察指标评价了青霉素菌渣的营养特性,并进一步分析了更有利于动物吸收利用的小分子氨基酸含量。结果表明:青霉素菌渣具有较高的潜在营养特性,粗蛋白、粗脂肪和总糖含量均较高,分别为568.767、82.919、320.51 mg/g,同时各种为生命体生长所需的氨基酸含量较丰富,其中苯丙氨酸、蛋氨酸含量最高。     

一株有脱硫性能细菌的分离与特性研究

报道了从含硫土壤中分离到的一株无机化能自养型的脱氮硫杆菌(Thiobacillus denitrificans)菌株L_1。该菌细胞呈球杆状,0.3～0.5μm×1.0～1.5μm,单根极生鞭毛运动,革兰氏染色阴性。可利用硫代硫酸盐、硫酸盐作为能源,在细胞内、外储存硫粒,在好气或厌气条件下能以硝酸盐为氮源,最适生长pH6.0。在约1％接种量时,该菌7d可使硫代硫酸盐和硫酸盐的脱硫率,分别达到34.37％和19.79％。     

Remediation of a chlorinated aromatic hydrocarbon in water by photoelectrocatalysis

Photoelectrocatalysis driven by visible light offers a new and potentially powerful technology for the remediation of water contaminated by organo-xenobiotics. In this study, the performance of a visible light-driven photoelectrocatalytic (PEC) batch reactor, applying a tungsten trioxide (WO₃) photoelectrode, to degrade the model pollutant 2,4-dichlorophenol (2,4-DCP) was monitored both by toxicological assessment (biosensing) and chemical analysis. The bacterial biosensor used to assess the presence of toxicity of the parent molecule and its breakdown products was a multicopy plasmid lux-marked E. coli HB101 pUCD607. The bacterial biosensor traced the removal of 2,4-DCP, and in some case, its toxicity response suggests the identification of transient toxic intermediates. The loss of the parent molecule, 2,4-DCP determined by HPLC, corresponded to the recorded photocurrents. Photoelectrocatalysis offers considerable potential for the remediation of chlorinated hydrocarbons, and that the biosensor based toxicity results identified likely compatibility of this technology with conventional, biological wastewater treatment.     

Restoring biochemical activity and bacterial diversity in a trichloroethylene-contaminated soil: the reclamation effect of vermicomposted olive wastes

Background, aim, and scope  In this work, the potential for using olive-mill solid waste as an organic amendment for biochemical and biological restoration of a trichloroethylene-contaminated soil, which has previously been stabilized through vermicomposting processes, has been explored. Materials and methods  Trichloroethylene-contaminated water was pumped into soil columns with a layer of vermicompost at 10-cm depth (biobarrier system). The impacts of the trichloroethylene on the microbial community were evaluated by determining: (1) the overall microbial activity (estimated as dehydrogenase activity) and enzyme activities related to the main nutrient cycles (β-glucosidase, o-diphenoloxidase, phosphatase, urease, and arylsulphatase activities). In addition, isoelectric focusing of the soil extracellular humic-β-glucosidase complexes was performed to study the enzymatically active humic matter related to the soil carbon cycle. (2) The soil bacterial diversity and the molecular mechanisms for the bacterial resistance to organic solvents were also determined. For this, polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) was used to detect changes in bacterial community structure and PCR-single-strand conformational polymorphism (SSCP) was developed and optimised for detection and discrimination of the resistance-nodulation-division (RND) genes amplified from the contaminated soils. Results  Vermicompost reduced, with respect to the unamended soil, about 30% of the trichloroethylene leaching during the first month of the experiment. Trichloroethylene had a marked negative effect on soil dehydrogenase, β-glucosidase, urease, phosphatase, and arylsulphatase activities. Nevertheless, the vermicompost tended to avoid this toxic effect. Vermicompost also displays stable humic-β-glucosidase complexes that increased the extracellular activity related to C-cycle in the contaminated soils. The isoelectric focusing technique showed a more biochemically active humic matter in the soil sampled under the vermicompost. The behaviour of the three main phyla of bacteria isolated from the DGGE bands was quite different. Bands corresponding to Actinobacteria disappeared, whereas those affiliated with Proteobacteria remained after the trichloroethylene contamination. The disappeared Actinobacteria became visible in the soil amended with the vermicompost. Bands corresponding to Bacteriodetes appeared only in columns of contaminated soils. In this study, six types of RND proteins were detected by PCR-SSCP in the natural soil, three in the trichloroethylene-contaminated soil and 7/5 in trichloroethylene-contaminated soil above/below the vermicompost in the biobarrier columns. Trichloroethylene tended to reduce or eliminate all the clones detected in the uncontaminated soil, whereas new efflux pumps appeared in the biobarrier columns. Discussion  Although enzymes incorporated into the humic substances of vermicomposted olive wastes are quite stable, trichloroethylene also inhibited the background levels of the soil extracellular β-glucosidase activity in the amended soils. The decrease was less severe in the biobarrier system, but in any case, no relation was found between the levels of trichloroethylene in soil and extracellular β-glucosidase activity, or between the latter and the quantity of humic carbon in soils. The isoelectric focusing technique was carried out in the humic fraction to determine whether the loss of activity occurred in overall extracellular β-glucosidase or in that linked to stable humic substances (humic–enzyme complexes). The contaminated soils showed the lower enzyme activities, whereas contaminated and amended soils presented greater quantity of focalised (and therefore stable) humic carbon and spectra heterogeneity: very different bands with higher enzyme activities. No clear relationship between trichloroethylene concentration in soil and diversity of the bacterial population was noted. Similar patterns could be found when the community structures of bacteria and microbial activity were considered. Since the use of the dehydrogenase assay has been recognised as a useful indicator of the overall measure of the intensity of microbial metabolism, these results could be attributed to PCR-DGGE methodology, since the method reveals the presence of dominant populations regardless of their metabolic state. Trichloroethylene maintained or even increased the number of clones with the DNA encoding for RND proteins, except for the contaminated soil located above the vermicompost. However, the main effect of trichloroethylene was to modify the structure of the community in contaminated soils, considering the type of efflux pumps encoded by the DNA extracted from soil bacteria. Conclusions  Trichloroethylene inhibited specific functions in soil and had a clear influence on the structure of the autochthonous bacterial community. The organic matter released by the vermicomposted olive waste tended to avoid the toxic effect of the contaminant. Trichloroethylene also inhibited the background levels of the soil extracellular β-glucosidase activity, even when vermicompost was present. In this case, the effect of the vermicompost was to provide and/or to stimulate the humic-β-glucosidase complexes located in the soil humic fraction >10⁴, increasing the resistance of the enzyme to the inhibition. The bacterial community from the soil presented significantly different mechanisms to resistance to solvents (RND proteins) under trichloroethylene conditions. The effect of the vermicompost was to induce these mechanisms in the autochthonous bacterial community and/or incorporated new bacterial species, able to grow in a trichloroethylene-contaminated ambient. Coupled biochemical and molecular methodologies are therefore helpful approaches in assessing the effect of an organic amendment on the biochemical and biological restoration of a trichloroethylene-contaminated soil. Recommendations and perspectives  Since the main biochemical and biological effects of the organic amendment on the contaminated soil seem to be the incorporation of biochemically active humic matter, as well as new bacterial species able to grow in a trichloroethylene-contaminated ambient, isoelectric focusing and PCR-SSCP methodologies should be considered as parts of an integrated approach to determine the success of a restoration scheme.     

Biodegradable dissolved organic carbon shapes bacterial community structures and co-occurrence patterns in large eutrophic Lake Taihu

Interactions between dissolved organic matter (DOM) and bacteria are central in the biogeochemical cycles of aquatic ecosystems; however, the relative importance of biodegradable dissolved organic carbon (BDOC) compared with other environmental variables in structuring the bacterial communities needs further investigation. Here, we investigated bacterial communities, chromophoric DOM (CDOM) characteristics and physico-chemical parameters as well as examined BDOC via bioassay incubations in large eutrophic Lake Taihu, China, to explore the importance of BDOC for shaping bacterial community structures and co-occurrence patterns. We found that the proportion of BDOC (%BDOC) correlated significantly and positively with the DOC concentration and the index of the contribution of recent produced autochthonous CDOM (BIX). %BDOC, further correlated positively with the relative abundance of the tryptophan-like component and negatively with CDOM aromaticity, indicating that autochthonous production of protein-like CDOM was an important source of BDOC. The richness of the bacterial communities correlated negatively with %BDOC, indicating an enhanced number of species in the refractory DOC environments. %BDOC was identified as a significant stronger factor than DOC in shaping bacterial community composition and the co-occurrence network, suggesting that substrate biodegradability is more significant than DOC quantity determining the bacterial communities in a eutrophic lake. Environmental factors explained a larger proportion of the variation in the conditionally rare and abundant subcommunity than for the abundant and the rare bacterial subcommunities. Our findings emphasize the importance of considering bacteria with different abundance patterns and DOC biodegradability when studying the interactions between DOM and bacteria in eutrophic lakes.     

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.     

Soil contamination with antibiotics in a typical peri-urban area in eastern China: Seasonal variation, risk assessment, and microbial responses

The prevalence and persistence of antibiotics in soils has become an emerging environmental issue and an increasing threat to soil security and global public health. The problem is more severe in areas undergoing rapid urbanization; however, the ecological risks of antibiotics, seasonal variability, and associated soil microbial responses in peri-urban soils have not been well-explored. The seasonal soil sampling campaigns were conducted in a typical peri-urban watershed in eastern China to investigate distribution of antibiotics. The results demonstrated higher mean concentrations of most antibiotic compounds in winter than in summer in peri-urban soils. The seasonal variations of norfloxacin, enrofloxacin, and ciprofloxacin were more significant than those of other antibiotics, due to their higher migration ability and bioavailability. An ecological risk assessment demonstrated that chlortetracycline, ciprofloxacin, doxycycline, and ofloxacin can pose high risks to soil microorganisms. Furthermore, the coexistence of multiple antibiotics obviously poses higher risks than individual compounds. A redundancy analysis demonstrated that tetracyclines mainly showed negative correlations with Firmicutes and Chloroflexi, and quinolones showed obviously negative correlations with Acidobacteria, Gemmatimonadetes, and Nitrospirae, suggesting potential inhibition from antibiotics on biological activities or biodegradation processes. However, the persistence of antibiotics in soil results in a significant decrease in bacterial diversity and a change in dominant species. Our results provide an overview of the seasonal variability of antibiotics and the associated effects on bacterial communities in peri-urban soils. The results can provide scientific guidance on decreasing soil contamination with antibiotics to enhance soil security in similar areas.     

A simple method to harness DC (direct current) electrical energy generated by bacterial activity

We developed a new system for collecting electrons generated by bacterial activity to supply DC (direct current) electrical energy. This system used eight titanium sub-electrodes (surface area of each sub-electrode: 189 cm²) connected to one central main titanium electrode (surface area of electrode: 1571 cm²). The distance between each sub-electrode and the main electrode was 30 cm. In an initial experiment, we collected electrons during composting cattle excreta, which was mixed with a commercially available microbial compost activator/starter. We analysed the relationships between the composting temperature and electrical current and voltage. Electrons were effectively collected and used successfully to obtain DC electrical energy. Generation of a stable voltage of approximately 0.5 V was clearly observed. This voltage was not related to compost temperature; however, the generated electrical current increased and decreased with compost temperature. The method was then used in a second experiment to collect electrons generated by bacterial activity in an agricultural field (area: 48 m²). Electrons in the field were effectively collected, and we measured a stable voltage of approximately 1.1 V and a stable current of approximately 0.7 mA. The system has the advantages of being simple, easily maintained, inexpensive, and applicable to large-scale agricultural fields.