一株新型鱼腥藻溶藻细菌的分离鉴定及其溶藻特性

蓝藻水华在全球范围频繁暴发,鱼腥藻(Anabaena)水华出现频度仅次于微囊藻(Microcystis)水华,其中的一些鱼腥藻种类能产生多样的藻毒素或异味物质,鱼腥藻水华去除技术的研发备受关注.本研究以从武汉市新洲区土壤中分离筛选到的一株细菌WP为材料,通过分析其生理生化特征及16S r DNA序列对其进行分类地位鉴定,考察了该菌株对真紧密鱼腥藻(Anabaena eucompacta)CHAB 1799的溶藻特性.结果表明,菌株WP与Lysinibacillus fusiformis NRS-350的同源性达到了99.6%,故鉴定WP属于Lysinibacillus属.菌株WP对真紧密鱼腥藻CHAB 1799发生溶藻作用的细菌阈值为6×10~3cfu/ml,其生理状态不会明显影响溶藻效果,且对不同生长时期的藻液均有较强的溶藻作用.该菌株的溶藻方式为间接溶藻.本研究结果丰富了溶藻细菌的资源库,也为治理鱼腥藻水华提供了更多的技术支持.     

微囊藻水华附生菌的代谢特征及对微囊藻生长的影响

为研究附生细菌对蓝藻水华细胞微环境的影响,从微囊藻水华中分离出34株藻附生细菌,研究其产酶能力.对氮代谢相关的酶活性测定结果显示:其中13株菌具有蛋白酶活性,5株菌具有较强的氨氮脱除能力,1株菌具有强烈的硝酸还原酶活性;4株菌具有较高的碱性磷酸酶活性;11株菌具有产脂酶的活性.通过菌和藻共同培养的方法,观察菌对微囊藻生长的影响,结果显示,有11株菌表现出较明显的促藻生长作用,占总筛选菌株总数的32%;有3株菌表现出较显著的抑制藻生长的作用,占总筛选菌株总数的9.9%.附生菌产酶能力与生长相关性的分析显示,促进微囊藻生长的附生菌都具有蛋白酶或脂酶活性.     

太湖磷转化细菌与水体磷形态关系

太湖水体中不同形态磷含量与磷转化细菌的关系研究结果显示:太湖水体中总磷和活性磷的含量分别为0.113mg,L和O.O11mg/L;无机磷和有机磷分解菌在底泥中达6.73x103cells/g,而在水体中仅为71cells/m1,且存在明显的时间和空间差异;根据菌落形态特征,分离筛选了3株有代表性的无机磷转化菌和7株占优势的有机磷分解菌.3tt无机磷转化菌经鉴定分别与巨大芽孢杆菌(Bacillus megaterium)、假单孢菌(Pseudomonas sp.)和类芽孢杆菌(Paenibacillus sp.)比较接近.而7株有机磷分解菌则分别与为苏云金芽孢杆菌(Bacillus thurigiensts)、短小芽孢杆菌(Bacillus pumilus)、芽孢杆菌(Bacillus SP.)、无芽孢杆菌(Bacterium sp.)、氧化微杆菌(Microbacterium oxydans)腊状芽孢杆菌(Bacillus cereus)、简单芽孢杆菌(Bacillus simples)接近;太湖水体中磷分解细菌主要归属于芽孢杆菌属(Bacillus)和假单孢菌属(Pseudomonas),对细菌降解性能进行研究的结果显示:无机磷分解细菌对太湖水体活性磷的贡献显著大于有机磷分解细菌的贡献率.     

我国水华蓝藻的新类群——阿氏浮丝藻(Planktothrix agardhii)生理特性

阿氏浮丝藻(Planktothrix agardhii)是水华蓝藻的重要类群,实验对我国不同省份分离到的5株阿氏浮丝藻生长速率、色素组成、光合活性参数进行了测定.结果表明分离于广州的HABll28叶绿素a(Chla)含量较低,藻蓝蛋白(c-Pc)比例和含量较高.电子传递速率(ETR)测定结果表明,HABIl28的ETR曲线较高,最大ETRme值也显著高于其余4株,高效的光能电子传递链弥补了光合色素Chla的不足,以致HABIl28保持了中性的生长速率.分离于武汉东湖的HAB631则与之正好相反,较高的ChLa受限于光能电子传递链的薄弱,生长速率表现较低.分离于北京、上海、昆明的3株藻株在各项生理指标上无显著差异.     

一株来源于水库的浮游蓝丝藻SK3株的生长及产毒特性

浮游蓝丝藻是水华蓝藻的常见种属,但对其生长和产毒特性认识不多本文首次对我国水体中的浮游蓝丝藻进行了研究.从广东省一重要供水水库中分离到一株丝状蓝藻,经形态特征鉴定为阿氏浮游蓝丝藻,并在不同的光照和温度下对该藻株进行了培养.结果表明,适合于该藻生长的光强和温度分别为20μmol/(m~2·s)和30℃;该藻株具有mcyE基因,能产生微囊藻毒素MCRR,产量为0.06μg/mg(DW).     

广东省千灯湖拟柱孢藻(Cylindrospermopsis raciborskii)的形态和产毒能力的株间差异及系统进化

拟柱孢藻(Cylindrospermopsis raciborskii)是一种入侵蓝藻,能够产生拟柱孢藻毒素(cylindrospermopsin,CYN),严重危害人类健康.现有研究表明该藻已在我国广泛分布,更是华南地区常见的水华蓝藻,但目前对其产毒和起源还缺乏了解.本研究以中国广东省千灯湖分离出的10株拟柱孢藻为材料,对它们的生长、形态和产毒特性进行了观测,并构建了基于nif H和rpo C1基因的双基因系统进化树.结果表明,10株拟柱孢藻藻丝体形态均呈笔直型,除QDH1藻株外,其他的拟柱孢藻均可产生端生异形胞,厚壁孢子也常有出现;10藻株丝体的平均长度在41.0~77.7μm之间,宽度在2.433~3.125μm之间,它们的长宽比值差异显著.10藻株的比生长速率差异极显著,为0.075~0.174 d-1.检测发现10株藻中仅QDH7藻株可检测到6个CYN合成酶基因,液相色谱串联质谱法(LC-MS/MS)分析表明该藻株主要产生deoxy-CYN异构体,其浓度可达1745.19 ng/mL.系统进化分析表明千灯湖的拟柱孢藻与澳大利亚、欧洲和国内其他地区的藻株同源性较高,但基于rpo C1和nif H基因不能区分千灯湖的产毒藻株和非产毒藻株.本研究证实了广东省水体中存在产毒拟柱孢藻,需防范这类新型产毒蓝藻水华所带来的生态风险.     

微囊藻胶鞘多糖和水溶性胞外多糖的化学特性

微囊藻(Microcystis)产生大量胞外多糖(EPS),包括包裹在细胞外的胶鞘多糖(CPS)和释放到周围环境中的水溶性多糖(RPS).为探究EPS在蓝藻水华发生中的生理生态学意义,迫切需要了解微囊藻EPS的化学特性.本文从太湖分离群体微囊藻,经过大约18个月实验室培养后,其中一些藻株转变为单细胞形态.选择5株群体藻和4株单细胞藻,比较分析这些藻株EPS的化学特性发现:(1)所有9株藻的EPS均为含有脱氧己糖的酸性杂多糖;(2)所有9株藻的CPS的糖醛酸含量(1.2%~2.1%)均低于RPS的糖醛酸含量(2.4%~6.2%);(3)所有9株藻的EPS均含有乙酰基和硫酸基,其中,每一株藻CPS的乙酰基含量均高于RPS的乙酰基含量,所有群体藻CPS的乙酰基含量(4.1%~6.6%)高于所有单细胞藻CPS的乙酰基含量(2.0%~3.2%).本文进而讨论了EPS化学特性对EPS水溶性和微囊藻群体形成的影响,以及对其生态学作用的影响.在这些化学特性中,乙酰取代基团被认为可能是影响微囊藻EPS生理生态学作用的重要因素.     

汾河太原河段水华藻类分类及分子系统研究

2012-2016年,每年的春、夏、秋季对汾河太原河段进行浮游植物样品采集.通过对样品的形态观察和描述,共鉴定出5种水华优势种,5月发生的裸藻水华优势种为裸藻属的膝曲裸藻(Euglena geniculata)和血红裸藻(E.sanguinea).而7-9月发生的微囊藻水华优势种为微囊藻属的铜绿微囊藻(Microcystis aeruginosa)、挪氏微囊藻(M.novacekii)和惠氏微囊藻(M.wesenbergii).分离纯化共得到11株单克隆水华藻,其中铜绿微囊藻8株,挪氏微囊藻2株,血红裸藻1株.运用cpcBA-IGS、gyrB和cpSSU基因序列构建分子系统发育树,进一步确定水华藻的系统分类地位,结果表明cpcBA-IGS是研究汾河太原河段铜绿微囊藻分类很好的分子标记,而cpSSU基因可很好地区分血红裸藻和其他裸藻种.     

中国蓝藻的新记录属——双囊藻属(Geminocystis)及分类和生态学讨论

现代分子生物学技术为蓝藻的生物多样性研究提供了更多信息和证据.单细胞蓝藻——双囊藻属（Geminocystis）是2009年从集胞藻（Synechocystis）分离出来的新属.2017年5月从青海湖的封闭子湖——月牙湖采集分离到一株单细胞蓝藻藻株,16S rRNA基因序列显示此藻株和双囊藻的藻种显示出最高的相似性,都在97%以上,并且在分子系统树聚为一束.此株的细胞结构以及超微结构显示,细胞纵轴具有窝式结构以及类囊体不规则的排列等双囊藻属所具有的特征.双囊藻是我国蓝藻的新报道纪录属,本研究还对双囊藻的分类和生态进行了讨论.     

转MT-like基因衣藻的重金属结合能力与抗性特征分析

通过对转MT－like基因衣藻和其野生品系的比较分析,发现转基因藻株对重金属镉和铜的结合能力及抗性均有大幅提高。在重金属结合能力方面,低浓度（5－10μmol/L）的镉溶液中,转基因藻重金属结合量是野生株的1.5倍左右,而相对高浓度（30－50μmol/L）的镉溶液中,由于重金属对藻细胞的毒害作用,使两者的差异不明显。在重金属铜和镉的抗性,转基因藻表现出明显高于野生藻株的抗性特征。用转基因藻去处理含铜的工业废水,结果发现转基因藻的处理效率可以提高10％以上。这说明DNA重组技术对提高藻类的废水处理效率方面具有较大的潜力。     

Microcystis,Raphidiopsis raciborskii and Dolichospermum smithii,toxin producing and non-toxigenic cyanobacteria in Yezin Dam,Myanmar

Yezin Dam is a man-made reservoir located close to Yezin village in Myanmar. Its water is used for irrigation, domestic purposes and as drinking water for many urban communities in the watershed area. In recent years, increased pollution due to the concurrent development around the dam has led to water quality deterioration. No detailed study on the distribution of cyanobacteria and toxin production has been conducted so far. In order to provide insight into the extent of cyanobacteria and cyanotoxins in the dam, water samples were collected once in January 2014 for the isolation of cyanobacterial strains and eight times between March 2017 and June 2018 for the investigation of physical, chemical and biological parameters. A total of 99 phytoplankton taxa belonging to 50 genera were recorded from Yezin Dam. Microscopic examination showed that a Dolichospermum sp. was the dominant cyanobacterium followed by small numbers of Microcystis, and Raphidiopsis raciborskii in all samples throughout the sampling period. 15 isolated cyanobacterial strains were classified morphologically and phylogenetically as Dolichospermum smithii, R. raciborskii and Microcystis and tested for microcystins (MCs), cylindrospermopsins (CYNs), saxitoxins (STXs) and anatoxins (ATXs) by liquid chromatography-tandem mass spectrometry (LC–MS/MS) and enzyme-linked immunosorbent assay (ELISA). The toxin analysis of all isolated Dolichospermum strains by ELISA and LC–MS did not indicate the presence of ATXs, STXs, CYNs nor MCs. Four of the five isolated Raphidiopsis strains produced CYN and deoxyCYN. One of the isolated Microcystis strains (AB2017/08) from Yezin Dam produced 22 MC congeners. Concentrations of 0.12 μg L⁻¹ CYNs and 0.34 μg L⁻¹ MCs were also found in an environmental sample from Yezin Dam by ELISA. The potential therefore exists for the use of untreated water from Yezin Dam to cause harmful effects on humans, domestic and wild animals.     

江西柘林湖水华蓝藻——长孢藻(Dolichospermum)的形态多样性及其分子特征

长孢藻是具有产多种蓝藻毒素及异味物质潜在能力的丝状异形胞蓝藻.为了探究长孢藻的生物学特性,我们从江西柘林湖分离24株长孢藻,经形态鉴定分成4个种类,分别为浮游长孢藻(Dolichospermum planctonicum)、近亲长孢藻(D. affine)、卷曲长孢藻(D. circinale)、螺旋长孢藻(D. spiroides). 16S rRNA基因序列表明柘林湖长孢藻与日本等地区的长孢藻高度相似,但只有近亲长孢藻与日本近亲长孢藻在分子系统进化树上聚为一支.基于16S-23S rRNA之间高变异性的ITS序列分析,4种长孢藻的D1-D1'螺旋结构相同,藻丝为直型的浮游长孢藻、近亲长孢藻与藻丝为弯型的卷曲长孢藻、螺旋长孢藻的Box-B、V3螺旋结构差异较大.通过对毒素和异味的分子生物学检测显示24株长孢藻均不产毒,但是4株浮游长孢藻及1株卷曲长孢藻含有土腥素合成基因.本研究不仅对柘林湖水华蓝藻的多样性以及潜在生态风险提供科学的基础资料,也在流域层面上为鄱阳湖的水生态系统研究和保护提供了一定的科学基础.     

Antifouling activity of commercial biocides vs. natural and natural-derived products assessed by marine bacteria adhesion bioassay

Biofilm formation is a key step during marine biofouling, the natural colonization of immersed substrata, leading to major economic and ecological consequences. Consequently, bacteria have been used for the screening of new non-toxic antifoulants: the adhesion of five strains isolated on three French locations was monitored using a fluorescence-based assay and toxicity was also evaluated. Nine biocides including commercial, natural and natural-derived products were tested. The commercial antifoulants, TBTO and Sea Nine showed low EC₅₀ but high toxicity. The non-commercial products TFA-Z showed significant anti-adhesion activities and appeared to be non-toxic, suggesting a specific anti-adhesion mechanism. In addition, the strains could be classified depending on their sensitivity to the molecules used even if strain sensitivity also depended on the molecules tested. In conclusion, TFA-Z would be a promising candidate as non-toxic antifoulant and our results strengthen the need to perform antifouling bioassays with a panel of strains showing different response profiles.     

江汉平原江陵地区9ka以来古气候演化的沉积记录

从一起自然发病的中华绒螯蟹(Eriocheir sinensis H．Milne-Edwards)肝胰腺中,分离做纯培养的6株(HQ010516A-1至HQ010516A-6)鳗利斯顿氏菌(Listonella anguillarum),进行了形态特征、主要理化特性等方面的检验．同时择代表菌株(HQ010516A-1)测定了16S rRNA基因序列、构建了系统发育树．另外,以代表菌株(HQ010516A-1)对健康蟹进行了人工感染试验,结果表明了该菌株在所检病例的病原学意义;用37种抗菌类药物对3株菌所做的药敏试验,结果均表现对头孢噻肟等29种药物呈现敏感或高度敏感(抑菌圈直径在16-40 mm)、对头孢唑啉等3种药物呈低度敏感(抑菌圈直径在10-12 mm)、对青霉素G等5种药物耐药(无抑菌圈形成)．     

Isolation and characterization of two crude oil-degrading yeast strains, Yarrowia lipolytica PG-20 and PG-32, from the Persian Gulf

Among six crude oil-degrading yeasts that were isolated from an oil-polluted area in the Persian Gulf, two yeast strains showed high degradation activity of aliphatic hydrocarbons. From an analysis of 18S rRNA sequences and biochemical characteristics, these strains were identified as Yarrowia lipolytica strains PG-20 and PG-32. Gas Chromatography (GC) analysis of the crude oil remaining in the culture medium after 1 week at 30°C showed that the strains PG-20 and PG-32 degraded 68% and 58% of crude oil, respectively. The optimal growth condition and biodegradation of hydrocarbons was in ONR medium with an acidic pH (pH5). These two strains may degrade aliphatic hydrocarbons more efficiently than aromatic hydrocarbons, although strain PG-20 had better degradation than strain PG-32. The two Y. lipolytica strains reduce surface tension when cultured on hydrocarbon substrates (1% v/v). These strains showed a cell surface hydrophobicity higher than 70%. These results suggested that Y. lipolytica strains PG-20 and PG-32 have high crude oil degrading activity due to their high emulsifying activity and cell hydrophobicity. In conclusion, these yeast strains can be useful for the bioremediation process in the Persian Gulf and decreasing oil pollution in this marine ecosystem.     

Prevalence and genetic profiles of Escherichia coli from mangroves and mangrove associated foods off Goa,India

A total of 120 samples comprising of water (45), sediment (45) and mangrove originated food (30) collected from mangrove ecosystems of Goa were screened for Escherichia coli employing ISO-16654 method. Seventy-one (59.16%) samples were positive for E. coli. The E. coli isolates were further characterized by serotyping, virulence gene profiling and pulsed field gel electrophoresis (PFGE). Water and sediment samples were analyzed for physico-chemical parameters. The serotypes reported were O1, O10, O13, O17, O36, O41, O50, O68, O105, O116, O141, O148, O159, O162 and rough types while, 23 strains could not be typed. The stx1 and stx2 genes were detected in 33(46.47%) and 16(22.53%) isolates, respectively. The XbaI restriction digestion patterns of the stx positive strains were diverse. Interestingly, few strains isolated from diarrheal patients and from water, sediment and food from mangrove sources were genetically similar. The study showed that the mangrove ecosystem could be a potential reservoir for pathogenic E. coli.     

Isolation and characterization of crude-oil-degrading bacteria from the Persian Gulf and the Caspian Sea

Twenty-five crude-oil-degrading bacteria were isolated from oil-contaminated sites in the Persian Gulf and the Caspian Sea. Based on a high growth rate on crude oil and on hydrocarbon degradation ability, 11 strains were selected from the 25 isolated strains for further study. Determination of the nucleotide sequence of the 16S rRNA gene showed that these isolated strains belonged to genera Acinetobacter, Pseudomonas, Gordonia, Rhodococcus, Cobetia, Halomonas, Alcanivorax, Marinobacter and Microbacterium. Among the 11 isolates, strains BS (Acinetobacter calcoaceticus, 98%) and PG-12 (Alcanivorax dieselolei, 98%) were the most effective in degrading crude oil. Rate of crude-oil degradation of 82% (isolate BS) and 71% (isolate PG-12) were observed after 1 week of cultivation in mineral medium. These strains had high emulsification activity and biosurfactant production. GC-MS analysis showed that A. dieselolei PG-12 can degrade different alkanes in crude oil. Screening of the distribution of the alkane hydroxylase gene in 25 isolates in relation to the source of isolation indicated that the group (II) alkane hydroxylase is prevalent in the Caspian Sea, but in the Persian Gulf, the frequency of the group (III) alkane hydroxylase gene is greater than that of the group (II) alkane hydroxylase gene.