徐闻8种珊瑚共附生真菌的分离及初步鉴定

从徐闻珊瑚保护区的8种珊瑚中分离共附生真菌,以获得珊瑚共附生可培养真菌的多样性信息。选用5种培养基,采用平板涂布法培养分离共附生真菌,并进行形态鉴定。8种珊瑚共分离121株真菌,鉴定了其中97株真菌,分属于14个属,青霉属和枝孢霉为优势种属,其次是曲霉属和木霉属。二异角孔珊瑚上分离出22株,数量最多;盔型珊瑚上分离菌株数量最少仅8株。除优势种属外,不同珊瑚样品上真菌种类分布有较大差异,炭角菌属和葡萄穗霉菌属均来源于盔形珊瑚,镰刀菌属在盔形珊瑚和角孔珊瑚上较为常见,盔形珊瑚样品上还分离出2株散囊菌属,1株节菱孢属,因此盔形珊瑚的种群最为丰富;不同培养基分离真菌的能力不同,淀粉培养基分离的菌株最多占所有分离菌株的30.6%,CDA培养基最少,但种类特异。     

具有多种胞外酶的对虾肠道黏附菌的分离和鉴定

用对虾饲料培养基从健康凡纳滨对虾肠道分离出500株黏附细菌,以产淀粉酶、脂肪酶和蛋白酶能力为指标,筛选出产该3种消化酶的细菌90株,占总菌株的18%.对其中生长较快的69株进行16S rDNA 基因测序,确定其分类地位.结果显示,69株菌分别属于不动杆菌属(Acinetobacter)、芽孢杆菌属(Bacillus)、葡萄球菌属(Staphylococcus)、假交替单胞菌属(Pseudoalteromonas)、气单胞菌属(Aeromonas)、嗜盐单胞菌属(Halomonas)、利斯顿氏菌属(Listonella)、莫拉氏菌属(Moraxella)等,其中数量最多是芽胞杆菌属,占鉴定细菌总数的53.62%,数量最少是气单胞菌属和嗜盐单胞菌属,均占鉴定细菌总数的2.90%.表明对虾肠道黏附菌群中具有较多能分泌多种消化酶的细菌,可进一步开发为促进对虾消化功能的益生菌     

苯扎溴铵复方消毒液在水产品加工厂的应用

用苯扎溴铵复方消毒液对大肠杆菌、金黄色葡萄球菌和枯草杆菌黑色变种芽胞进行定量杀菌、现场模拟杀菌及金属腐蚀试验。结果表明 :含量为 0 .2、0 .4 g/L的苯扎溴铵复方消毒液分别对大肠杆菌和金黄色葡萄球菌在消毒时间 1min以上有完全杀菌效果 ;含量为 0 5g/L的苯扎溴铵复方消毒液对枯草杆菌黑色变种芽胞在消毒 15min以上有完全杀菌效果 ;含量为 0 .2g /L的苯扎溴铵复方消毒液对水产品加工厂中生产人员的双手、手套和装虾的塑料筛上的大肠杆菌在消毒1min以上达到完全杀菌效果 ;苯扎溴铵复方消毒液对不锈钢、碳钢和铜无腐蚀 ,对铝只有轻微的腐蚀。     

苯扎溴铵复方消毒液在水产品加工厂的应用

用苯扎溴铵复方消毒液对大肠杆菌、金黄色葡萄球菌和枯草杆菌黑色变种芽胞进行定量杀菌、现场模拟杀菌及金属腐蚀试验。结果表明：含量为0．2、0．4g／L的苯扎溴铵复方消毒液分别对大肠杆菌和金黄色葡萄球菌在消毒时间1min以上有完全杀菌效果；含量为0．5g／L的苯扎溴铵复方消毒液对枯草杆菌黑色变种芽胞在消毒15min以上有完全杀菌效果；含量为0．2g／L的苯扎溴铵复方消毒液对水产品加工厂中生产人员的双手、手套和装虾的塑料筛上的大肠杆菌在消毒1min以上达到完全杀菌效果；苯扎溴铵复方消毒液对不锈钢、碳钢和铜无腐蚀，对铝只有轻微的腐蚀。     

海洋短小芽孢杆菌ZH1-6菌株的鉴定及生物学特性

从新鲜近江牡蛎中分离到一株拮抗细菌,命名为ZH1-6,该菌对金黄色葡萄球菌、乙型副伤寒沙门氏菌及大肠杆菌等多种病原菌有较强的抑制作用。通过形态学观察、常规生理生化指标测定、16SrDNA序列测定和同源性分析,鉴定该菌株为短小芽孢杆菌。研究表明:ZH1-6能利用多种碳、氮源,其生长温度范围为10-50℃,生长pH范围为pH3-10,最适初始pH为6.0。     

海洋土曲霉C23-3与不同类型海洋微生物共培养对其次生代谢产物的影响

【目的】研究海洋土曲霉C23-3与不同类型海洋微生物共培养对其次生代谢产物的影响,挖掘土曲霉菌株C23-3的次生代谢潜力。【方法】将副溶血弧菌、枯草芽孢杆菌和胶红酵母的活菌或灭活菌体分别以不同时间与土曲霉C23-3共培养;采用DPPH自由基清除法和Ellman法分别评价各提取物的抗氧化和乙酰胆碱酯酶抑制活性,采用滤纸片法比较各提取物的抗菌活性,采用HPLC指纹图谱分析各提取物化学成分的差异。【结果与结论】发现不同类型的微生物与土曲霉共培养均可激活一些原本沉默的活性次级代谢产物;菌株及共培养物的化学拮抗能力、HPLC指纹图谱中的特征代谢产物变化揭示在活菌培养体系中土曲霉占竞争优势;而且灭活菌体中可能存在某些信号物质,在12 d时对土曲霉群体自感应信号分子丁内酯-I的产生具有比活菌更强的刺激作用。研究结果为深入挖掘海洋真菌代谢潜力及抗老年痴呆等活性先导化合物的发现提供基础。     

鱼干曲霉菌分离株鉴定及产毒特性

【目的】探明湛江市售鱼干中曲霉菌(Aspergillus spp.)分离株产AFB₁的特性。【方法】采用马铃薯-葡萄糖琼脂(PDA)对鱼干中的曲霉菌进行分离纯化,根据菌落形态、显微形态观察和ITS序列分析对分离株进行鉴定,采用液相色谱-串联质谱(LC-MS/MS)法分析菌株的AFB₁合成量。【结果】鱼干中曲霉菌的分离率高达47.66%,经形态学鉴定发现以黄曲霉(Aspergillus flavus)A03、烟曲霉(Aspergillus fumigatus)H32、杂色曲霉(Aspergillus versicolor)M23、欧式曲霉(Aspergillus europaeus)M11、棘孢曲霉(Aspergillus aculeatus)B21等为主,其中红笛鲷干(Lutjanus sanguinaus)中黄曲霉菌的含量高达1200 CFU/g。黄曲霉A03在红笛鲷干培养基中常温(27~30℃)培养21 d,产AFB₁能力达到3.8 ng/mL。【结论】鱼干中存在产AFB₁的黄曲霉菌,警示鱼干中存在AFB₁污染的风险。     

低值鱼蛋白多肽-铁(Ⅱ)螯合物的酶解制备及其抗氧化、抗菌活性

以南海低值鱼蛋白为原料,采用木瓜蛋白酶和风味酶的复合酶水解得到多肽水解物(HP),并制备多肽-铁(Ⅱ)螯合物,对螯合工艺及螯合产物的结果及抗氧化和抗菌功能特性进行了初步研究。结果表明:1)水解度为5%的多肽与铁(Ⅱ)螯合效果最佳,螯合率为94.15%;2)化学分析及红外光谱表明活性肽与亚铁离子螯合并形成稳定结构的螯合物;(3)经过50%乙醇(体积分数)沉淀的螯合铁具有明显的抗氧化活性,其抗氧化作用相当于维生素E的90%,经过80%乙醇(体积分数)沉淀的螯合物具有显著的抗枯草牙孢杆菌和金黄色葡萄球菌活性,其抑菌圈均直径为13mm。     

特呈岛陆地成年乔木资源的研究

对特呈岛陆地成年乔木资源进行调查与分析,结果表明,特呈岛陆地的成年乔木:第一类(胸径≥20cm)共1584株,隶属26科41属51个树种,优势树种突出,其中百株以上的树种有4种;第二类(胸径≥20cm与树高≥6m)共1445株,其中大乔木1株、中乔木368株、乔木1076株;第三类(树龄≥80a)30株,其中有古树3种共23株及古树后续资源7株,有胸径≥100cm的大树14株;第四类有二级国家重点保护野生树种樟树32株。建议调整古树后续资源规定标准为树龄≥50a,以加强成年乔木保护。     

湛江市古树名木资源调查与分析

调查了湛江城市建成区及城乡结合带的古树名木299株,其中古树292株,名木7株,分属 16科20属24种。树木种类以榕属(Ficus)植物最为显著,占总资源的77.9%;箭毒木(Antiaris toxicaria)、鹊肾树(Streblusasper)秋茄树(Kandeliacandel)、竹节树(Caralliabrachiata)、叉叶木 (Crescentiaalata)和牛蹄豆(Pithecellobiumdulce)等,为湛江特有的古树名木资源。分析了古树名 木资源的分布与湛江城市建设的关系,讨论了古树名木的生长状况,并提出保护建议。     

Phylogenetic diversity and bioactivity of culturable deep-sea-derived fungi from Okinawa Trough

Deep-sea sediments are now recognized as a home for rich and largely microbial community. Recently, it has been believed in an increasing number of studies that bacteria could be abundant in deepsea sediments of many types; however, fungi in deep-sea sediments remain relatively unknown. The phylogenetic diversity and bioactivity of culturable deep-sea-derived fungi from Okinawa Trough sediments were investigated in traditional method combined with fungal identification of molecular biology in this study. A total of 76 isolates belonged to 15 fungal taxa were recovered in a harsh condition of low nutrient and low temperature, indicating that the fungal communities in deep-sea sediments from Okinawa Trough were relatively abundant and diversified. Aspergillus, Cladosporium, and Penicillium were the dominant fungal genera, while Mycosphaerella, Purpureocillium, and Schizophyllum were relatively rare in the deep-sea sediments from Okinawa Trough. Among the six genera recovered, Mycosphaerella was firstly recovered from deep-sea sediments in this study. Moreover, about 75% of the extracts from the 15 fungal representative isolates displayed distinct bioactivity against at least one indicator bacterium or marine macrofouler, emphasizing the potentials of these deep-sea-derived fungi from Okinawa Trough as producers of bioactive metabolites. Notably, isolates Cladosporium oxysporum SCSIO z001 and Penicillium citrinum SCSIO z049 displayed a wide spectrum of bioactivities, isolates Cladosporium cladosporioides SCSIO z015, Cladosporium sphaerospermum SCSIO z030, and Penicillium verruculosum SCSIO z007 exhibited a strong anti-bacterial-growth activity, and isolate Penicillium chrysogenum SCSIO z062 displayed a strong anti-larval-settlement activity. These results suggest that these isolates deserved further study as potential sources of novel bioactive metabolites.     

化学诱导对2株海洋真菌次级代谢产物及其生物活性的影响

【目的】以2株海洋真菌为研究对象,探究不同种类及浓度的化学表观遗传修饰剂对其次级代谢产物化学多样性和生物活性的影响。【方法】基于化学表观遗传修饰策略,采用两种不同的化学表观遗传修饰剂5-氮杂胞嘧啶核苷(5-azaC)和丁酸钠,分别对Penicillium sp.019和Aspergillus terreus ZN4-5-4两株海洋来源真菌进行表观遗传修饰。通过观察真菌在培养基中的生长状态变化,并根据薄层层析(TLC)指纹图谱、高效液相色谱(HPLC)图谱、乙酰胆碱酯酶(AChE)抑制、1,1-二苯基-2-三硝基苯肼(1,1-Diphenyl-2-picryl-hydrazyl,DPPH)自由基清除和卤虫致死活性筛选模型等技术手段,从中筛选出代谢产物丰富、AChE抑制活性与抗氧化活性产物丰富且毒性小的发酵培养条件。【结果】添加1 mmol/mL的丁酸钠诱导剂可使菌株019和ZN4-5-4的次生代谢产物的种类增加,且同时能提高该菌株次级代谢产物的AChE的抑制活性和DPPH自由基清除活性。因此,确定菌株019和ZN4-5-4的最优发酵条件均是添加浓度为1 mmol/mL丁酸钠的PDB培养基。【结论】化学表观遗传修饰策略指导下,添加诱导剂5-azaC或者丁酸钠对2株海洋真菌次生代谢产物的产生有影响。     

Inhibition of norsolorinic acid accumulation to Aspergillus parasiticus by marine actinomycetes

Thirty-six strains of marine actinomycetes were isolated from a sample of marine sediment collected from the Yellow Sea and evaluated in terms of their inhibitory activity on the growth of Aspergillus parasiticus and the production of norsolorinic acid using dual culture plate assay and agar diffusion methods. Among them, three strains showed strong antifungal activity and were subsequently identified as Streptomyces sp. by 16S rRNA gene sequencing analysis. The supernatant from the fermentation of the MA01 strain was extracted sequentially with chloroform and ethyl acetate, and the activities of the extracts were determined by tip culture assay. The assay results show that both extracts inhibited mycelium growth and toxin production, and the inhibitory activities of the extracts increased as their concentrations increased. The results of this study suggest that marine actinomycetes are biologically important for the control of mycotoxins, and that these bacteria could be used as novel biopesticides against mycotoxins.     

Antimicrobial Activity of Ulva reticulata and Its Endophytes

Seaweeds are known to exhibit various antimicrobial properties,since it harbours an enormous range of indigenous bioactive compounds.The emergence of drug resistant strains has directed to the identification of prospective metabolites from seaweed and its endophytes,thereby exploiting the properties in resisting bacterial diseases.The current study was aimed to assess the antimicrobial activity of extracts obtained from Ulva reticulate,for which metabolites of Ulva reticulata and its endophytes were extracted and assessed against human pathogens like Staphylococcus aureus,Escherichia coli,Pseudomonas aeruginosa,Salmonella typhi,and Bacillus subtilis.It was observed that the hexane extract of isolate VITDSJ2 was effective against all the tested pathogens but a significant inhibition was observed for Staphylococcus aureus and Escherichia coli.Further,Gas chromatography coupled with Mass spectroscopy(GC-MS) revealed the existence of phenol,3,5-bis(1,1-dimethylethyl) in the crude hexane extract which is well-known to possess antibacterial activity.The effective isolate VITDSJ2 was identified to be the closest neighbour of Pseudomonas stutzeri by phenotypic and genotypic methods.The crude extracts of the seaweed Ulva reticulata was also screened for antibacterial activity and the hexane extract was effective in showing inhibition against all the tested pathogens.The compound in the crude extract of Ulva reticulata was identified as hentriacontane using GC-MS.The extracts obtained from dichloromethane did not show significant activity in comparison with the hexane extracts.Hence the metabolites of Ulva reticulata and the bacterial secondary metabolites of the endophytes could be used in the treatment of bacterial infections.     

Isolation and characterization of bioactive fungi from shark Carcharodon carcharias' gill with biopharmaceutical prospects

Until recently,little was known about the fungi found in shark gills and their biomedicinal potential. In this article,we described the isolation,bioactivity,diversity,and secondary metabolites of bioactive fungi from the gill of a shark(Carcharodon carcharias). A total of 115 isolates were obtained and grown in 12 culture media. Fifty-eight of these isolates demonstrated significant activity in four antimicrobial,pesticidal,and cytotoxic bioassay models. Four randomly selected bioactive isolates inhibited human cancer cell proliferation during re-screening. These active isolates were segregated into 6 genera using the internal transcribed spacer-large subunit(ITS-LSU) rDNA-sequence BLAST comparison. Four genera,Penicillium,Aspergillus,Mucor,and Chaetomium were the dominant taxa. A phylogenic tree illustrated their intergenera and intragenera genetic diversity. HPLC-DAD-HRMS analysis and subsequent database searching revealed that nine representative strains produced diverse bioactive compound profiles. These results detail the broad range of bioactive fungi found in a shark's gills,revealing their biopharmaceutical potential. To the best of our knowledge,this is the first study characterizing shark gill fungi and their bioactivity.     

Antifouling activities of marine bacteria associated with sponge (Sigmadocia sp.)

The present study aimed at assessing the antifouling activity of bacteria associated with marine sponges. A total of eight bacterial strains were isolated from the surface of sponge Sigmadocia sp., of them, SS02, SS05 and SS06 showed inhibitory activity against biofilm-forming bacteria. The extracts of these 3 strains considerably affected the extracellular polymeric substance producing ability and adhesion of biofilm-forming bacterial strains. In addition to disc diffusion assay, microalgal settlement assay was carried out with the extracts mixed with polyurethane wood polish and coated onto stainless steel coupons. The extract of strain SS05 showed strong microalgal settlement inhibitory activity. Strain SS05 was identified as Bacillus cereus based on its 16S rRNA gene. Metabolites of the bacterial strains associated with marine invertebrates promise to be developed into environment-friendly antifouling agents.     

Preliminary study on PAH degradation by bacteria from contaminated sediments in Xiamen Western Sea,Fujian, China

In order to estimate the biodegradation of three polycyclic aromatic hydrocarbons (PAHs) compounds, bacterial strains were isolated from marine sediments in three heavily contaminated sites (Yuandang Lake, Dongdu Port and Aquacultural zones in Maluan Bay) in Xiamen Western Sea. The results show three bacterial strains, which used pyrene as the sole carbon source, were identified as strains of Aureobacterium sp.,Arthrobacter sp., Rhodococcus sp. The PAH-degrading bacteria isolated had a strong ability to degrade phenanhrene, fluoranthene and pyrene at different degradation rates. The highest degradation rate was observed when three PAH compounds were mixed with an individual strain in the medium. The three PAHs were degraded after one week with a degradation rate of 89.94 % for phenanthrene and 93.4 % for both of fluoranthene and pyrene.In addition, after 25 days of incubation, the degradation rate was 99.98 % for phenanthrene and 99.97 % for both of fluoranthene and pyrene. Optical density was measured to estimate bacterial growth during the degradation of PAHs. Highest levels of bacterial growth were observed with a three PAH mixture in the culture, suggesting that the concentration of PAHs influenced bacterial growth and the highest levels of degradation for most series were detected after one week of incubation.     

石油污染物的微生物降解

从炼油厂污水池底泥中富集、驯化、分离、筛选,得到4种优势石油降解菌。采用摇床培养,研究了各优势菌和混合菌对石油烃的降解性能。结果表明:4种菌和混合菌20 d可将初始质量浓度为10000 mg/L的石油烃依次降解90.8%、88.9%、57.8%、49.8%、91.2%;培养液中石油烃的半衰期依次为5.5、6、15、19、5 d。初步鉴定4种菌分别属:节细菌(Arthrobacter sp.)、芽胞杆菌(Bacillus sp.)、不动杆菌(Acinetobacter sp.)、不动杆菌(Acinetobacter sp.)。