Improvement of production of lipopeptide antibiotic iturin A using fish protein

To enhance the production of lipopeptide antibiotic iturin A, nutrient contents of the culture mediums were investigated in both submerged and biofilm fermentations. As a carbon source maltose and as nitrogen source, fish protein was used. In submerged fermentation maltose uptake was found lower (12%) compared to biofilm fermentation (15%) that was associated with higher cellular growth in biofilm. However, requirement of nitrogen (fish protein) concentration was found similar in both submerged and biofilm fermentations. Production of iturin A in submerged fermentation with 12% maltose and 5% fish protein was 4450 mg/L, and in biofilm fermentation it was 5050 mg/L when 15% maltose and 5% fish protein was used.     

Biofilm formation and lipopeptide antibiotic iturin A production in different peptone media

Biofilm fermentation is a newly developed promising technique in fermentation technology. In this study no.3 and no.3S media have been used for the lipopeptide antibiotic iturin A production by Bacillus subtilis RB14. The main component of no.3 and no.3S media is Polypepton and Polypepton S, respectively. B. subtilis RB14 produces thick stable biofilm and high amount of iturin A in no.3S medium. Whereas, impaired biofilm formation and lower iturin A production was observed in no.3 medium. From the analytical information it was observed that the amounts of metal ions, such as K⁺, Ca²⁺ and Mn²⁺, cysteine and cellulose are lower in Polypepton compared to the Polypepton S. To investigate their effect on biofilm formation and iturin A production cysteine, cellulose, K⁺, Ca²⁺ and Mn²⁺ were added respectively into the no.3 medium at similar amount that Polypepton S contains. It was observed that individual addition of K⁺, Ca²⁺, cysteine and cellulose had no effect on biofilm formation, cellular growth induction or iturin A production. However, when Mn²⁺ was supplemented in no.3 medium, biofilm development was restored with an improved production of iturin A. Finally, combined addition of investigated substances into the no.3 medium resulted with highly folded, thick biofilm with high cellular growth and iturin A production compared to the original no.3 medium.     

Solid state fermentation of lipopeptide antibiotic iturin A by using a novel solid state fermentation reactor system

A new solid state fermentation reactor (SSFR) for solid substrate was used for the production of lipopeptide antibiotic iturin A using Bacillus subtilis RB14-CS. Solid state fermentation (SSF) is the technique of cultivation of microorganisms on solid and moist substrates in the absence of free water. SSF has shown much promise in the development of several bioprocesses and products because of their several advantages like absence of free water that allows simplified downstream processing and low cost. SSFR allows agitation of the SSF culture with improved temperature control and air supply. Interestingly, when okara, the widely available waste product from the tofu industries, was used as the solid substrate for the SSFR, no iturin A production was observed. However, without agitation, production of iturin A was observed in the SSFR but the production level remained low. The low production of iturin A was found to be due to the heat generation and excess temperature rise inside the reactor system during the fermentation process. Maintaining the temperature within a range of 25–30°C, production of iturin A was significantly improved in the SSFR. This was comparable to the laboratory scale production, and signifies the potential application of the SSFR for SSF.     

脂肽类生物表面活性剂产生菌的分离及特性研究

从石油污染土壤中分离筛选获得一株产生生物表面活性剂菌株Y8A,经生理生化实验、16S rDNA序列分析等将其鉴定为芽孢杆菌属（Bacillus sp.）.Y8A能在22h内将发酵液的表面张力从68.3mN·m-1降到23.5 mN·m-1.经TLC和傅立叶红外光谱分析, 菌株Y8A产生的生物表面活性剂为脂肽类.20mg·L-1 Ca2+和Fe2+能显著促进其生长和表面活性剂的产生;菌株Y8A在20～30℃,pH 5～12范围内产生表面活性剂的能力较强;LB培养基中添加1%乳糖对生长的影响不大,但能够明显促进Y8A产生生物表面活性剂,而葡萄糖、蔗糖抑制表面活性剂的产生.Y8A能够促进石油降解菌Y1D和F11对石油的降解和功夫菊酯降解菌ZZH对功夫菊酯的生物降解.     

升级A/O工艺污水处理系统中抗生素抗性基因的分布与去除研究

针对北方某采用升级A/O工艺的生活污水处理厂,使用实时荧光定量PCR技术,探究污水厂中ARGs的分布及各处理工艺段对ARGs的去除效果.结果表明:四环素抗性基因(tetA、tetC和tetM)、磺胺抗性基因(sul1和sul2)、大环内酯抗性基因(ermA和ermF)和喹诺酮抗性基因(parC和gyrA)在污水和污泥中均被检出.污水厂进水中ARGs的绝对丰度为2.65×10³～1.01×10⁶ copies·mL^-1,升级A/O工艺未能有效削减ARGs,出水中ARGs的绝对丰度为9.22×10³～1.15×10⁶ copies·mL^-1,污泥中ARGs的绝对丰度为8.07×10⁷～2.65×10¹¹ copies·g^-1.深度处理工艺对ARGs的去除效率对比结果显示,生物活性炭工艺对ARGs的削减效果优于紫外消毒.     

固体发酵提高水生植物发酵产物蛋白含量的研究

采用微生物固体发酵技术对伊乐藻(Elodea nuttalli)、苦草(Vallinmeria spiralis)和喜旱莲子草(Alterranthera philoxerides)3种高等水生植物进行单细胞蛋白生产的研究,分析单一菌种和混合菌种发酵过程中水生植物粗蛋白含量和纤维素酶活的变化过程.研究结果表明,与单一霉菌发酵相比,利用霉菌与酵母混合发酵,可明显提高发酵产物粗蛋白产量,其中以黑曲霉(Aspergillus niger)与产朊假丝酵母(Candida utilis)混合发酵苦草的粗蛋白含量最高,产物中粗蛋白含量最高可达39.88%,粗蛋白增加率为84.2%,使其有可能成为鱼、家禽和家畜的蛋白饲料来源.因此,利用固体发酵处理水生植物可以实现水生植物的资源化.     

基于三维成像技术的污泥菌群中抗生素抗性基因转移动态特征研究

应用活细胞三维成像技术,实时和原位观测了活性污泥菌群中抗生素抗性基因转移动态特征.微流控芯片是一种细菌生长的良好载体,活细胞成像系统低光毒性不损伤细菌活性,两者结合能有效实现基因转移的长时间原位观测,能够区分抗性基因的水平转移和垂直转移.结果表明:垂直转移是污泥菌群中抗性基因的主要转移方式,菌群纵向转移速率与其厚度有关,菌群内部转移率较高,基因转移速率为0.24～0.29 h~(-1).     

A/O生物膜法强化处理石化废水及生物膜种群结构研究

采用A/O生物膜反应器处理石化综合废水.反应器在O段添加装有改性聚氨酯泡沫的多孔塑料球载体,强化有机物的降解效率.反应器进水分别为水解酸化池出水(阶段I),石化工业废水与生活污水比例为3:1(阶段II)以及单纯的石化工业废水(阶段III). 结果表明,尽管进水COD和氨氮波动较大,但出水COD和氨氮的去除率保持稳定,说明生物膜反应器具有较好的抗冲击负荷能力.在HRT为30h, COD和氨氮的去除率为74%~77%和96%~93%,总氮和总磷的去除率为58%和79%.第II阶段进水为工业废水和生活污水混合的处理效果最好,出水COD和氨氮浓度分别为(63±12)mg/L和(0.75±0.28)mg/L.出水总氮主要为硝酸氮,亚硝酸氮的浓度很低(小于0.1mg/L),表明硝化作用进行得较为完全.进水中有机物的分子量主要分布在小于1kDa(70.9%)和大于100kDa(10.4%).出水中大于10kDa的有机物所占比例减小,分子量主要分布在小于1kDa(56.6%)和1~5kDa(26.2%),表明A/O生物膜反应器对大分子有机物的降解较好.454高通量测序结果表明: 生物膜中变形菌门菌群所占比例最大(60.0%),其次是浮霉菌门(16.9%)和拟杆菌门(9.8%).在属的水平检测到氨氧化菌(AOB)Nitrosomonas和亚硝酸盐氧化菌(NOB) Nitrospiraceae Nitrospira以及反硝化菌Azospira和Thermomonas.NOB的比例较高,这与反应器较好的硝化作用相一致.     

A preliminary investigation on the occurrence and distribution of antibiotic resistance genes in the Beijiang River, South China

The occurrence of antibiotic resistance genes (ARGs) was investigated and quantified in 20 water samples collected in the Beijiang River, South China. Sulfonamide- and tetracycline-resistant bacteria were present in 17 and 14 of the collected 20 samples. For sulfonamide ARGs, sulI and sulII were frequently observed in the Beijiang River. The levels of sulI were higher than sulII (p < 0.05), with the mean values of (1.41 ± 1.12) × 10-2 and (1.58 ± 1.71) × 10-3 copies/16S rDNA, respectively. For tetracycline ARGs, tetG had the highest frequency, 100%, followed by tetA (85%), tetO (85%), tetC (70%), tetX (60%), tetM (40%) and tetQ (20%), while tetE and tetS were not detected in all the samples from the Beijiang River. On the other hand, tetC had the highest concentration, ranging from 8.30 × 10-2 to 13.20 copies/16S rDNA. The poor correlation between ARGs and antibiotic concentrations revealed that the self-amplification and persistence of ARGs were the reasons that made ARGs exist in the water environment even though the antibiotic selecting pressure was absent. Because so few field measurements have been conducted for investigating the levels of ARGs in rivers in South China, this study provides an important insight on better understanding the occurrence and spread of ARGs in such an ecosystem.     

磺胺甲唑对海水养殖废水处理过程中抗性细菌及抗性基因的富集作用

抗生素在海水养殖过程中大量使用,但仅有少部分被生物体利用,含有抗生素的废水进入水处理系统后,抗生素、抗性菌和抗性基因的响应过程尚不完全清楚.应用缺氧/好氧移动床生物膜反应器（A/O-MBBR）处理含磺胺甲唑（SMX）的海水养殖废水,探究在SMX选择压力下,反应器内抗生素和抗性基因丰度的变化规律,以及微生物群落和可培养的抗性细菌种群的响应.结果表明,在进水SMX浓度为500 μg·L^-1,水力停留时间为8 h,SMX加入初期会对NH₄⁺-N和NO₂^--N的去除率有轻微影响,随后逐步恢复;同时去除约32%的SMX,且78%以上SMX在缺氧区完成;抗性基因在缺氧区富集明显高于好氧区,在缺氧区磺胺类抗性基因（sul1）绝对丰度上升2.43 log,磺胺类抗性基因（sul2）上升1.71 log;而在好氧区,sul1 绝对丰度上升1.17 log,sul2 上升0.91 log.抗性平板培养结合高通量测序表明,假交替单胞菌属（Pseudoalteromonas）在反应器可培养抗性细菌中占最优势.高通量测序分析发现可培养的抗性细菌假单胞菌属（Pseudomonas）在反应器内占比最高.表明含SMX的海水养殖废水可促进水中抗性基因的富集,部分抗性细菌的数量显著增加.     

猪场废水生产微生物絮凝剂发酵特性及动力学特征

以猪场废水作为原材料生产微生物絮凝剂,考察了发酵过程中溶解氧(DO)、COD、氨氮、总磷及pH值的变化规律,利用Logistic和Luedeking-Piret模型对絮凝菌生长和代谢产物生成的动力学过程进行了拟合.结果显示,菌体的生长在0~60h是对数生长期,12~36h菌体快速生长,菌体干重、细胞浓度OD600和菌落数分别由0.09g/L,0.26和1.3×107cfu迅速增加到0.76g/L,0.58和3.5×107cfu,稳定期(66h)菌落数达到5×107cfu.絮凝菌代谢产物的主要絮凝活性成分是蛋白质,与菌体生长呈相关型.猪场废水中磷源含量充足,作为缓冲液,基本维持了絮凝菌发酵过程中pH值的恒定.此外,Logistic和Luedeking-Piret模型的拟合结果能够较好地描述絮凝菌生长和代谢产物生成的动力学过程.     

耦合发酵产氢尾液处理的微生物电化学系统研究

区别于传统的稀释或加缓冲剂调节pH值的方法,本文提出采用微生物电解池（MEC）电调控暗发酵尾液pH值,并进一步采用微生物电合成系统（MES）降解废液产甲烷.结果表明,在MEC处理产氢暗发酵尾液过程中,伴随着阴极侧氢气的产生,暗发酵尾液中大量H⁺被消耗,溶液pH值从4.5升高到8.7;随后在MES中,产氢发酵尾液中有机物被进一步降解产生甲烷,其平均产甲烷速率达到4.5mmol/（L·d）,且在21d内化学需氧量（COD）去除率达到89%,远优于没有经过pH调控的产氢发酵尾液MES中的产甲烷性能.     

低温下AHLs对多级A/O工艺中好氧生物膜特性的影响

针对低温条件下多级A/O工艺出水氨氮容易超标问题,进行外源添加酰基高丝氨酸内酯类信号分子(AHLs)对低温下多级A/O工艺好氧段生物膜特性的影响研究.结果 表明:添加100nmol/L的己酰基高丝氨酸内酯(C6-HSL)后生物膜硝化效率提高了22.34％,十二烷酰基高丝氨酸内酯(C12-HSL)则将生物附着量提高了24...     

低负荷进水的改良型A2／O曝气池的改造研究

工业集聚区都配套相关的污水处理厂,但是很多工业集聚区,在刚刚起步的时候配套的污水处理厂都面临配套管网建设滞后、收集废水量较少等问题。焦作市万方工业集聚区污水处理厂进水单一,生化性差,进水负荷低于设计值,曝气池连续曝气造成污泥解絮,间歇曝气导致污泥在反应区发生沉淀,为解决上述问题,在好氧区添加潜水搅拌器,能够合理控制溶解氧,防止污泥解体,并且还可以避免生物反应区污泥发生沉淀现象,降低了电耗,出水指标好,达到了设备改造预期效果。     

多段进水A/O生物膜工艺运行性能的研究

为提高多段进水A/O生物膜工艺的脱氮效率,按照进入各缺氧池的COD量与硝态氮量的比值相同,且等于一最优比值的原则进行流量分配,按照容积负荷相等的原则设计各反应单元,对多段进水A/O生物膜工艺的流量分配和反应单元容积分配同时进行优化,并采用三段进水A/O生物膜反应器试验了等流量分配模式下运行和优化模式下运行的出水水质的差异.试验结果表明,当多段进水A/O生物膜工艺在两种模式下采用相同参数运行时,优化模式下总氮(TN)去除率明显高于等流量模式,其分别为88.8%和80.3%.优化模式与等流量分配模式对COD和总凯氏氮的去除率差别不大(均可达到97%和98%以上).采用该优化设计方法可以显著提高该工艺的TN去除效率.     

Pregnancy-associated plasma protein A: A possible marker in the classification and prenatal diagnosis of Cornelia de Lange syndrome

The concentration of human placental lactogen (hPL), pregnancy specific beta-1 glycoprotein (SP-1) and pregnancy-associated plasma protein A (PAPP-A) were analysed in consecutive serum samples from a patient who gave birth to a child with Cornelia de Lange syndrome. HPL and SP-1 were present in normal concentrations from week 20 to week 35 of gestation whereas PAPP-A could not be detected in any of the samples examined. Immunohisto-chemical examination of two placentae from Cornelia de Lange syndrome revealed normal localization of hPL and SP-1 but the absence of PAPP-A from the syncytiotrophoblast. The significance of association between Cornelia de Lange syndrome and compromised synthesis of PAPP-A is discussed.     

A/O-MBBR工艺处理生活污水性能及菌群结构

采用设计规模为100m³/d的一体化缺/好氧-移动床生物膜反应器(A/O-MBBR)处理实际生活污水,通过265d的中试研究考察了该工艺在多因素扰动下的除碳和脱氮性能,并对不同运行阶段微生物群落结构的动态变化进行了研究.结果表明,一体化A/O-MBBR系统具有良好的COD去除效果和脱氮性能.当好氧池溶解氧(DO)浓度和进水碳氮比(COD/N)分别为2.5~3.5mg/L和(7.9±2.0)时,COD、NH₄⁺-N和TN去除率分别达到(93.3±5.4)%、(99.1±0.6)%和(67.9±10.5)%.Proteobacteria、Bacteroidetes和Chloroflexi在不同运行时期均有较高的相对丰度,保证了有机物的高效去除.A/O-MBBR系统脱氮功能菌在运行初期主要分布于活性污泥中,且相对丰度较低.长期运行后,生物膜与活性污泥中均同时检出大量硝化菌和反硝化菌.其中,相对丰度最高的硝化菌为Nitrospira,主要分布于生物膜上(19.48%~28.05%).反硝化菌则以Thauera、Terrimonas和Dokdonella等为主.     

厨余发酵液作为中试A/O-MBR外增碳源的脱氮特性

利用厨余发酵液作为A/O-MBR的外增碳源,考察在不同发酵液投加量和水力停留时间(HRT)条件下的脱氮性能以及膜污染特点.结果表明,发酵液的反硝化速率和COD利用率与乙酸钠接近,比葡萄糖高.通过A/O-MBR的运行发现,厨余发酵液作为碳源能够增强微生物活性,强化反硝化过程,从而提高脱氮效率.TN的去除效率随着发酵液投加量的增加而增大.发酵液投加前后,反应器内的EPS并没有积累,膜污染速率变化较小.化学清洗可以有效地去除膜污染物质,恢复膜通量.同时,延长HRT有助于提高脱氮效率和缓解膜污染.     

Prenatal diagnosis of charcot-marie-tooth disease type 1a (CMT1A) using molecular genetic techniques

Charcot-Marie-Tooth disease type 1A (CMT1A) is a frequent hereditary motor and sensory neuropathy of the peripheral nerves. In most cases, the disease is associated with a 1.5 Mb tandem duplication at 17p11.2. A 42-year-old pregnant woman requested prenatal diagnosis because of her age and since both her husband and two children were severely affected with CMT1. The CMT1A duplication was demonstrated in the father's, the two children's, and the fetus's DNA using different molecular genetic methods. Although cytogenetical analysis showed a normal female karyotype in the fetus, the parents decided to terminate the pregnancy because of the genetic risk associated with the CMT1A duplication.     

A simple fluorescence anisotropy assay for detection of bisphenol A using fluorescently labeled aptamer

Bisphenol A(BPA) is one of the environmental endocrine disruptors(EDCs), and BPA contamination in environment can cause high risks to human health. Rapid determination of BPA on sites is in high demand in environmental analysis. Taking advantage of aptamers as affinity ligands and fluorescence anisotropy(FA) analysis, we developed a simple and rapid FA assay for BPA by employing a single tetramethylrhodamine(TMR) labeled short 35-mer DNA aptamer against BPA. The assay is based on the BPA-binding...