微生物制药菌渣管理信息平台设计研究

针对我国微生物制药菌渣监管体系不完善的问题,提出采用GIS、GPS、GPRS、计算机网络通信和数据处理等技术手段,对菌渣的收集、储存、转运、处理处置等关键环节进行动态实时监控,进而构建菌渣管理信息平台,并给出了相应的管理制度建议,对平台的规划以及系统的功能结构进行了一些探索,提出了微生物制药菌渣事故响应制度和菌渣生产和处理企业奖惩制度,有助于促进制药菌渣全过程管理的有效实现。     

甾体类药物发酵菌渣的无害化与资源化研究进展

我国为甾体类原料药的生产大国,微生物转化法生产甾体药物的过程中产生的发酵菌渣是一种非常宝贵的资源,但发酵菌渣中仍存在少量有毒的甾体类残留物,在我国被列入《国家危险废物名录》。目前,甾体药物发酵菌渣的处理仍存在一定困难,严重制约着制药企业的健康发展,为此很多研究者针对药物发酵菌渣的无害化与资源化途径开展了广泛的研究。从甾体类药物发酵菌渣的来源、污染属性及国家相关管理要求等方面入手,分别综述了目前包括水热处理、碱热处理、微波处理等预处理技术,燃料化、肥料化、材料化等资源化利用技术,焚烧处置、安全填埋等处置技术在内的甾体药物发酵菌渣无害化与资源化方面的研究现状与最新进展,并对不同处理处置技术的可行性与可操作性进行了综合分析,最后对该领域未来的研究方向进行展望。     

青霉素菌渣理化特性及其资源化利用研究现状

青霉素菌渣是青霉素发酵工艺中产生的残余固体废弃物,其产生量大、易产生二次污染,利用过程中会造成环境风险,在一定程度上限制了制药企业的发展。对目前国内外青霉素菌渣的资源化和处置技术进行了综述与讨论,青霉素菌渣的青霉素残留效价脱毒是目前亟待解决的问题;此外对青霉素菌渣的综合利用过程中重点解决的问题进行了预测及展望。     

抗生素菌渣处理处置技术进展

我国是抗生素原料药生产大国,抗生素菌渣产生量大、处理难度大,且属于危险固废,其合理、安全处置是目前制药企业亟待解决的难题。对抗生素菌渣的焚烧、肥料化、饲料化、填埋、能源化及其他处理处置技术的现状进行了归纳和总结,并对抗生素菌渣处理处置技术的发展趋势进行了展望。     

泰妙菌渣肥制备及其对土壤的影响

文章采用热活化过硫酸钾氧化法对泰妙菌素菌渣进行无害化处理制得菌渣肥。采用土壤模拟方法研究了泰妙菌素菌渣肥对土壤中酶活性和微生物数量的影响。通过正交实验得到菌渣肥最佳制备条件:过硫酸钾的投加量为25 g/L,温度为60℃,含水率为85%,处理时间为30 min。此条件下,菌渣中截短侧耳素的去除率为92%。通过土壤模拟试验发现:菌渣肥对土壤酶活性具有一定的促进作用,且随着施用时间的延长,土壤酶活性逐渐恢复到原土状态。土壤真菌、细菌和放线菌数量略微出现波动之后逐渐恢复到原土中微生物数量状态。菌渣肥对土壤微生物数量的影响小于菌渣的影响,所以热活化过硫酸钾氧化法用于菌渣肥的制备是可行的。     

青霉素菌渣堆肥过程中青霉素钠降解菌的分离与鉴定

为了研究开发青霉素发酵菌渣堆肥资源化与无害化技术,采用传统的富集、分离、纯化等微生物学方法,在青霉素菌渣与猪粪混合堆肥过程中筛选出一株青霉素钠高效降解菌——PC-2,并对其进行形态表征和基于16S rRNA基因序列的微生物种属鉴定. 结果表明:菌株PC-2属螯合球菌属(Chelatococcus sp.),其能够利用青霉素钠为唯一碳源生长,但外加碳、氮源可显著提高菌株PC-2对青霉素钠的降解效率. 当葡萄糖为碳源、蛋白胨为氮源、菌株PC-2接种量为14%、pH为6~8时,菌株PC-2在37 ℃下振荡培养6 h,对初始ρ(青霉素钠)为400 mg/L的青霉素钠的降解率可达98%以上. 自堆肥过程中获取高效青霉素钠降解菌PC-2,预示着其在菌渣堆肥过程中的应用潜力,也有助于深入开展青霉素制药菌渣的安全有效与无害化处理处置方法的研究.      

林可霉素菌渣堆肥微生物群落多样性分析

本试验以林可霉素菌渣-猪粪为原料、污泥-猪粪堆肥作对照,研究了林可霉素菌渣堆肥过程中残留林可霉素的降解情况,并基于Illumina Mi Seq高通量测序分析了林可霉素菌渣堆肥过程中微生物菌群的变化.结果表明:通过堆肥处理可以大幅度降解林可霉素菌渣中残留的林可霉素,经过33 d的堆肥处理后,林可霉素的残留量从最初的1 800 mg·kg-1降到483mg·kg-1,降解率高达73%.同时高通量测序结果表明,由于高含量的林可霉素残留,在堆肥初期和高温期林可霉素菌渣堆肥中细菌群落的分布丰度和多样性指数均低于污泥-猪粪堆肥,但真菌群落丰度和多样性均高于污泥-猪粪堆肥.林可霉素菌渣堆肥中细菌主要以Paucisalibacillus、Cerasibacillus、Bacillus、Virgibacillus、Ureibacillus、Paenibacillus、Sinibacillus属为主,而污泥-猪粪堆肥中主要以Truepera、Actinomadura、Pseudosphingobacterium、Pseudomonas、Luteimonas、Ureibacillus属为主,两者堆肥中微生物群落结构存在显著差异.随着堆肥进入腐熟期,林可霉素残留大幅度降解,抗生素对微生物的胁迫减小或解除,林可霉素菌渣-猪粪堆肥和污泥-猪粪堆肥相比,无论是细菌还是真菌,其微生物群落已逐渐趋同.表明堆肥处理可以大幅降解林可霉素残留,增加微生物多样性,有利于实现林可霉素菌渣无害化处理和资源化利用.     

氨基糖苷类抗生素菌渣残留检测方法与资源化研究进展

文章对氨基糖苷类抗生素菌渣主要的检测方法以及资源化处理处置技术进行归纳和总结,为准确检测菌渣中抗生素残留及提高其无害化处理及资源化利用效率提供依据,最后促进抗生素制药企业的健康发展。     

青霉素菌渣肥对西红柿根际土壤微生物和酶活性的影响

将不同剂量的青霉素菌渣肥施入种植西红柿的土壤中,研究菌渣肥对西红柿根际土壤微生物和酶活性的影响。结果表明:随着菌渣肥施入量的增加,细菌、放线菌和真菌数量总体上均逐渐增加,脲酶活性变化不明显,蔗糖酶活性逐渐增大,过氧化氢酶活性先增加后降低。在整个生长期,施入菌渣肥的3个处理单元中细菌、真菌数量逐渐增加,放线菌数量逐渐减少;不同处理单元间的酶活性变化规律不同。相关性分析表明:部分微生物之间呈显著或极显著正相关,部分酶之间呈显著正相关,酶与微生物之间则呈显著或极显著正相关。青霉素菌渣肥能够有效地改善土壤肥力。     

我国抗生素菌渣资源化研究新进展

抗生素菌渣是一种宝贵资源,在我国也属于危险废物。由于缺乏成熟高效的处理与资源化技术,以及科学完善的安全性评估方法,抗生素菌渣的处理问题严重地制约了制药企业的健康发展。结合最新相关文献和研究成果,从抗生素菌渣处理与资源化技术、安全性评估两方面综述了目前抗生素菌渣资源化取得的最新进展,总结了抗生素处理与资源化技术的最新发展,并对不同资源化技术的可行性进行了分析,提出了以抗生素及其代谢产物残留评估和抗性基因评估为基础的安全性评估方法,并总结了评估指标实现的技术困难。     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.     

Effects of two sludge application on fractionation and phytotoxicity of zinc and copper in the soil

The potential harm of heavy metals is a primary concern in application of sludge to the agricultural land. A pot experiment was conducted to evaluate the effect of two sludges on fractionation of Zn and Cu in soil and their phytotoxicity to pakchoi. The loamy soil was mixed with 0%, 20%, 40%, 60% and 80% (by weight) of digested sewage sludge (SS) and composted sludge (SC). The additions of both sludges caused a significant raise in all fractions, resulting in that exchangeable (EXCH) and organic bound (OM) became predominance of Zn and organic bound Cu occupied the largest portion. There was more available amount of Zn and Cu in SS treatments than SC treatments. During the pot experiment, the concentration of Zn in EXCH, carbonate (CAR) and OM and Cu in EXCH and OM fractions decreased in all treatments, so their bioavailability reduced. Germination rate and plant biomass decreased when the addition rate was high and the best yield appeared in 20% mixtures at the harvest of pakchoi. The two sludges increased tissue contents of Zn and Cu especially in the SS treatments. Zn in pakchoi was not only in relationship to ΔEXCH and ΔCAR forms but also in ΔOM forms in the sludge-soil mixtures. Tissue content of Cu in pakchoi grown on SC-soils could not be predicted by ΔEXCH. These correlation rates between Zn and Cu accumulation in pakchoi and variation of different fractions increased with time, which might indicate that sludges represented stronger impacts on the plant in long-term land application.     

Intraspecific differences in effects of co-contamination of cadmium and

A hydroponic experiment was carried out to study intraspecific differences in the effects of different concentrations of cadmium (Cd)(0-10 mg/L) and arsenate (As(V)) (0-8 mg/L) on the growth parameters and accumulation of Cd and As in six wheat varieties Jing-9428, Duokang-1, Jingdong-11, Jing-411, Jingdong-8 and Zhongmai-8. The endpoints of wheat seedlings, including seed germination,biomass, root length and shoot height, decreased with increasing the Cd and As concentrations. Significant differences in seed germination, biomass, root length, shoot height and the accumulation of Cd and As were observed between the treatments and among the varieties (p ＜ 0.05). The lethal dosage 50% were about 20, 80, 60, 60, 80 and 20 mg As/L for Jing-9428, Duokang-1, Jingdong-11,Jing-411, Jingdong-8 and Zhongmai-8, respectively, and the corresponding values for Cd were about 30, 80, 20, 40, 60 and 10 mg Cd/L, respectively. Among the six varieties, Duokang-1 was found to be the most resistant to Cd and As toxicity, and Zhongmai-8 was the most sensitive to Cd and As co-contamination. The resistance of the six varieties was found dependant on the seedling uptake of Cd and As. Duokang-1 was the most suitable for cultivation in Cd and As co-contaminated soils.     

Removal of heavy metals from sewage sludge by low costing chemical method and recycling in agriculture

ＲｅｍｏｖａｌｏｆｈｅａｖｙｍｅｔａｌｓｆｒｏｍｓｅｗａｇｅｓｌｕｄｇｅｂｙｌｏｗｃｏｓｔｉｎｇｃｈｅｍｉｃａｌｍｅｔｈｏｄａｎｄｒｅｃｙｃｌｉｎｇｉｎａｇｒｉｃｕｌｔｕｒｅＷｕＱｉｔａｎｇ，ＮｙｉｒａｎｄｅｇｅＰａｓｃａｓｉｅ，ＭｏＣｅｈｕｉＦ...     

Single and joint effects of pesticides and mercury on soil urease

The influence of two pesticides including chlorimuron-ethyl and furadan and mercury （Hg） on urease activity in 4 soils （meadow burozem and phaeozem） was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase （up to 13%-21%） in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship （P〈0.05） between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose （ED50） values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations.     

Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.