凡纳滨对虾低盐度高产虾池环境微生物生态研究

对广东珠三角地区,凡纳滨对虾低盐度高产虾池环境微生物调查结果,养殖水体异养菌平均数量为5.15×10⁴cfu·mL^-1,致病性弧菌为5.00×10³cfu·mL^-1,池底泥浆中异养细菌平均数量为2.41×10⁶cfu·mL^-1,致病性弧菌为1.45×10⁵cfu·mL^-1。不同虾池异养细菌的数量差别小而稳定,致病性弧菌的数量差别及波动大。淤泥较深的老化虾池,水体和泥浆中异养细菌平均分别为6.10×10⁴cfu·mL^-1和2.89×10⁶cfu·mL^-1,致病性弧菌为6.00×10³cfu·mL^-1和2.14×10⁵cfu·mL^-1;无淤泥虾池水体和泥浆中异养细菌平均分别为4.53×10⁴cfu·mL^-1和2.08×10⁶cfu·mL^-1,致病性弧菌为4.34×10³cfu·mL^-1和9.86×10⁴cfu·mL^-1。老化虾池底泥致病性弧菌明显偏高,成为老化虾池易爆发虾病的重要原因。低盐度虾池水体异养菌和致病性弧菌的数量变化,表现为养殖前期高,中后期低而稳定;池底泥浆中异养菌和致病性弧菌的数量变化,呈不规则波动。高的养殖水温对养殖环境异养菌和致病性弧菌表现抑制作用,养殖环境微生物与水体浮游藻类也有一定的关系。     

Abundance of potentially pathogenic micro-organisms in Penaeus monodon larvae rearing systems in India

Monodon baculovirus (MBV), external fouling organisms (EFO) and bacteria (especially Vibrio species) were monitored during 1996–1997 at nine different Penaeus monodon rearing hatcheries in India. Total cultivable heterotrophic bacteria, Vibrio-like-bacteria, presumptive Vibrio harveyi, Vibrio anguillarum, Vibrio vulnificus counts were determined from shrimp eggs, post larvae, rearing tank water, source sea water, feed (Artemia nauplii and microencapsulated feed). The MBV infected post larvae and their environment showed higher Vibrio-like-bacteria than uninfected post larvae. An over-whelming predominance of presumptive Vibrio harveyi and Vibrio anguillarum was observed in post larval rearing tank water, MBV infected and uninfected post larvae. Vibrio-like-bacteria in Artemia nauplii clearly showed the possible source of these pathogenic bacteria in the hatchery environments. Quantitative analysis of Vibrio-like-bacteria in hatcheries revealed that when the Vibrio-like-bacteria increases to 2 × 10² CFU mortality of the post larvae occurs. Abundance of these micro-organisms in hatchery samples indicated that they are opportunistic pathogens which can invade the shrimp tissue, subsequently cause disease when the post larvae were under stressful conditions.     

植物-固定化菌剂联合修复多环芳烃污染土壤

以火凤凰根际土壤中发现的3种优势菌[分枝杆菌(Ⅰ)、产黄纤维单胞菌(Ⅱ)、少动鞘氨醇单胞菌(Ⅲ)]构建的多菌剂体系为供试菌剂,针对大港油田原油污染土壤,将固定化供试菌剂接种于修复植物火凤凰根际,探讨供试菌剂强化火凤凰修复多环芳烃(PAHs)污染土壤的效果。结果表明: 处理ⅠⅢ(有效活菌数为10⁹ cfu·mL^-1)和ⅠⅡⅢ(有效活菌数为10⁷ cfu·mL^-1)对PAHs的降解有促进作用,PAHs降解率分别为32.2%和41.4%,均显著高于相应对照处理。此外,处理ⅠⅡⅢ对火凤凰的地下生物量有明显促进作用,比对照处理增加了31.2%。表明由3种优势菌构建的多菌剂ⅠⅡⅢ可以作为火凤凰修复PAHs污染土壤的强化手段,为微生物强化植物修复技术提供了新的修复思路及方法。     

风车草(Cyperus alternifolius)人工湿地系统氮去除及氮转化细菌研究

研究了风车草人工湿地污水处理系统TN去除率及氮转化细菌的数量。结果表明:风车草人工湿地对TN的去除率为73.8%,与无植物人工湿地系统相比较,去除率提高了17.4%。风车草人工湿地氨化细菌为7.98×10⁵cfu·g^-1(细砂),硝化细菌为1.95×10⁵MPN·g^-1(细砂),反硝化细菌为5.89×10⁴1.95×10⁴MPN·g^-1(细砂)。与无植物系统氮转化细菌相比,氨化细菌无明显差异,硝化细菌及反硝化细菌均高出1个数量级。     

Microbial community structure in autotrophic nitrifying granules characterized by experimental and simulation analyses

This study evaluates the community structure in nitrifying granules (average diameter of 1600 μm) produced in an aerobic reactor fed with ammonia as the sole energy source by a multivalent approach combining molecular techniques, microelectrode measurements and mathematical modelling. Fluorescence in situ hybridization revealed that ammonia-oxidizing bacteria dominated within the first 200 μm below the granule surface, nitrite-oxidizing bacteria a deeper layer between 200 and 300 μm, while heterotrophic bacteria were present in the core of the nitrifying granule. Presence of these groups also became evident from a 16S rRNA clone library. Microprofiles of NH₄⁺, NO₂^–, NO₃^– and O₂ concentrations measured with microelectrodes showed good agreement with the spatial organization of nitrifying bacteria. One- and two-dimensional numerical biofilm models were constructed to explain the observed granule development as a result of the multiple bacteria–substrate interactions. The interaction between nitrifying and heterotrophic bacteria was evaluated by assuming three types of heterotrophic bacterial growth on soluble microbial products from nitrifying bacteria. The models described well the bacterial distribution obtained by fluorescence in situ hybridization analysis, as well as the measured oxygen, nitrite, nitrate and ammonium concentration profiles. Results of this study are important because they show that a combination of simulation and experimental techniques can better explain the interaction between nitrifying bacteria and heterotrophic bacteria in the granules than individual approaches alone.     

Oligotrophic properties of heterotrophic bacteria and in situ heterotrophic activity in pelagic seawates

Abstract The distribution of heterotrophic bacteria in polluted coastal and unpolluted pelagic seawaters was studied using a ¹⁴C-MPN method with either five of seven kinds of ¹⁴C-organic compounds as substrates. The total number of heterotrophic bacteria in pelagic waters ranged from 9.2 × 10³ to 5.4. ¢ 10⁴ cell/ml and more than 85% of the heterotrophic bacteria were represented by obligate oligotrophs. In coastal waters, the number of heterotrophs was one order of magnitude higher (av. 3.5 ¢ 10⁵ cells/ml), and eutrophic and facultatively oligotrophic bacteria were predominant. Oligotrophs in pelagic waters had a high specificity for the utilization of amino acids, especially glycine, and acetate-utilizing bacteria were scarce. The in situ maximum uptake rates of glutamate and glycine were much higher than those of glycolate and acetate. Acetate uptake rates were extremely low or not detectable in pelagic waters. The specificity of uptake kinetics is assumed to depend on the existence of obligate oligotrophs as dominant bacteria in pelagic seawater.     

NOTE ESTABLISHMENT OF AXENIC CULTURES OF ANABAENA FLOS-AQUAE AND APHANOTHECE NIDULANS (CYANOBACTERIA) BY LYSOZYME TREATMENT

A new method using lysozyme for the production of axenic cultures of Anabaena flos-aquae De Brebisson and Aphanothece nidulans P. Richter was developed. Cyanobacterial growth was not inhibited at concentrations up to 1.2 g·L⁻¹ of lysozyme, whereas the growth of heterotrophic bacteria was suppressed. At concentrations up to 0.8 g·L⁻¹ of lysozyme, ampicillin caused a reduction of heterotrophic bacteria. The axenic cultures of these strains were acquired through a simple treatment using 1.0 g·L⁻¹ of lysozyme without ampicillin. These cyanobacteria resisted digestion by lysozyme at our experimental concentrations, whereas bacteria were digested selectively. This method of purification seems to be especially useful with cyanobacterial species that are sensitive to antibiotics or other germicidal agents.     

Effects of nitrogen addition on soil respiration in shrublands in Mt. Dongling,Beijing, China

Aims Soil respiration from terrestrial ecosystems is an important component of terrestrial carbon budgets. Compared to forests, natural or semi-natural shrublands are mostly distributed in nutrient-poor sites, and usually considered to be relatively vulnerable to environmental changes. Increased nitrogen (N) input to ecosystems may remarkably influence soil respiration in shrublands. So far the effects of N deposition on shrubland soil respiration are poorly understood. The aim of this study is to investigate the soil respiration of Vitex negundo var. heterophylla and Spiraea salicifolia shrublands and their response to N deposition. Methods We carried out a N enrichment experiment in V. negundo var. heterophylla and S. salicifolia shrublands in Mt. Dongling, Beijing, with four N addition levels (N₀, control, 0; N₁, low N, 20 kg N·hm^-2·a^-1; N₂, medium N, 50 kg N·hm^-2·a^-1 and N₃, high N, 100 kg N·hm^-2·a^-1). Respiration was measured from 2012-2013 within all treatments.Important findings Under natural conditions, annual total and heterotrophic respiration were 5.91 and 4.23, 5.76 and 3.53 t C·hm^-2·a^-1 for the V. negundo var. heterophylla and S. salicifolia shrublands, respectively and both were not affected by short-term N addition. In both shrubland types, soil respiration rate exhibited significant exponential relationships with soil temperature. Temperature sensitivity (Q₁₀) of total soil respiration in V. negundo var. heterophylla and S. salicifolia shrublands ranged from 1.44 to 1.58 and 1.43 to 1.98, and Q₁₀ of heterotrophic soil respiration ranged from 1.38 to 2.11 and 1.49 to 1.88, respectively. Short-term N addition decreased only autotrophic respiration rate during the growing season, but had no significant effects on total and heterotrophic soil respiration in V. negundo var. heterophylla shrubland. In contrast, N addition enhanced the heterotrophic soil respiration rate and did not influence autotrophic and total soil respiration in S. salicifolia shrubland.     

亚热带红壤侵蚀区马尾松不同套种模式生态系统碳平衡

以福建省长汀县红壤侵蚀区马尾松低效林套种杨梅、无患子、油茶及黄栀子的改造模式林分为研究对象,对林分各组分生物量年净生长量、含碳率及土壤异养呼吸进行定位观测,分析套种模式对低效马尾松林分生态系统碳储量格局及碳平衡的影响.结果 表明:杨梅、无患子、油茶、黄栀子和马尾松不同器官含碳率的变化范围分别为41.1％～ 50.1％、...     

生物过滤法处理城市污水处理厂臭气

对某城市污水处理厂沉砂池的臭气进行了生物过滤处理。生物过滤池滤料层高1.5m,滤料体积55m³,臭气处理量为12000m³·h^-1。对除臭效果120d的监测和研究结果表明,进气中H₂S浓度在1.96～4.63mg·m^-3之间,NH₃的浓度为2.21～5.68mg·m^-3。处理后,出气中H₂S的浓度在0.03～0.97mg·m^-3间,去除率均达80%以上,NH₃浓度最高值仅为0.46mg·m^-3,低于1.0mg·m^-3的一级排放标准。生物除臭复合微生物主要包括细菌、酵母和霉菌,其中细菌含量最高,每克干滤料细菌含量达1.8～3.1×10⁸CFU。菌种鉴定结果表明,除臭菌主要有芽孢杆菌属(Bacillus)、假单胞菌属(Pseudomonas)、动胶菌属(Zoogloea)、不动杆菌属(Acinetobacter)、硫杆菌属(Thiobacillus)、酵母属(Saceharomyces)、假丝酵母属(Candida)、曲霉属(Aspergillus)、青霉属(Penicillium)、根霉属(Rhizopus)等种属的微生物。     

Purification and characterization of a Na/K dependent alginate lyase from turban shell gut Vibrio sp. YKW-34

An alginate lyase with high specific enzyme activity was purified from Vibrio sp. YKW-34, which was newly isolated from turban shell gut. The alginate lyase was purified by in order of ion exchange, hydrophobic and gel filtration chromatographies to homogeneity with a recovery of 7% and a fold of 25. This alginate lyase was composed of a single polypeptide chain with molecular mass of 60 kDa and isoelectric point of 5.5–5.7. The optimal pH and temperature for alginate lyase activity were pH 7.0 and 40 °C, respectively. The alginate lyase was stable over pH 7.0–10.0 and at temperature below 50 °C. The alginate lyase had substrate specificity for both poly-guluronate and poly-mannuronate units. The k_cat/K_m value for alginate (heterotype) was 1.7 × 10⁶ s⁻¹ M⁻¹. The enzyme activity was completely lost by dialysis and restored by addition of Na⁺ or K⁺. The optimal activity exhibited in 0.1 M of Na⁺ or K⁺. This enzyme was resistant to denaturing reagents (SDS and urea), reducing reagents (β-mercaptoethanol and DTT) and chelating reagents (EGTA and EDTA).     

Enhanced production of free trans-astaxanthin by oxidative stress in the cultures of the green microalga Chlorococcum sp

Free trans-astaxanthin accumulated in the alga Chlorococcum sp. was markedly enhanced from 3.664 mg g⁻¹ cell dry weight to 5.724 mg g⁻¹ cell dry weight when the culture was supplemented with hydrogen peroxide (0.1 mM) under mixotrophic conditions of growth. After saponification, a total of 7.086 mg astaxanthin per g cell dry weight was achieved. Similarly, in heterotrophic cultures, the total astaxanthin content was increased from 1.034 mg g⁻¹ cell dry weight without H₂O₂ to 1.782 mg g⁻¹ cell dry weight with 0.1mM H₂O₂. Results indicate that hydrogen peroxide effectively induces the formation of free trans-astaxanthin in Chlorococcum sp.     

Microbial characterization during composting of municipal solid waste

This study investigates the prevailing physico-chemical conditions and microbial community; mesophilic bacteria, yeasts and filamentous fungi, bacterial spores, Salmonella and Shigella as well as faecal indicator bacteria: total coliforms, faecal coliforms and faecal Streptococci, present in a compost of municipal solid waste. Investigations were conducted in a semi-industrial pilot plant using a moderate aeration during the composting process. Our results showed that: (i) auto-sterilization induced by relatively high temperatures (60–55°C) caused a significant change in bacterial communities. For instance, Escherichia coli and faecal Streptococci populations decreased, respectively, from 2×10⁷ to 3.1×10³ and 10⁷ to 1.5×10³ cells/g waste dry weight (WDW); yeasts and filamentous fungi decreased from 4.5×10⁶ to 2.6×10³ cells/g WDW and mesophilic bacteria were reduced from 5.8×10⁹ to 1.8×10⁷ bacteria/g WDW. On the other hand, the number of bacterial spores increased at the beginning of the composting process, but after the third week their number decreased notably; (ii) Salmonella disappeared completely from compost by the 25th day as soon as the temperature reached 60°C; and (iii) the bacterial population increased gradually during the cooling phase. While Staphylococci seemed to be the dominant bacteria during the mesophilic phase and at the beginning of the thermophilic phase, bacilli predominated during the remainder of the composting cycle. The appearance of gram-negative rods (opportunistic pathogens) during the cooling phase may represent a serious risk for the sanitary quality of the finished product intended for agronomic reuse. Compost sonication for about 3 min induced the inactivation of delicate bacteria, in particular gram-negatives. By contrast, gram-positive bacteria, especially micrococcus, spores of bacilli, and fungal propagules survived, and reached high concentrations in the compost.     

Vegetable juice aids the recovery of heated spores of non-proteolytic Clostridium botulinum

Heating spores of non-proteolytic Clostridium botulinum at 85C for 2 min followed by plating on a standard laboratory medium reduced the count of viable spores by a factor of greater than 10⁴. A similar result was obtained when the plating medium was supplemented with juice from courgette, carrot or mung bean sprout. When plating was on media supplemented with hen egg white lysozyme or juice from turnip, swede, flat bean, cabbage or potato, heating at 85C for 10 min did not reduce the viable count by a factor of 10⁴. Thus these vegetable juices increased the measured heat resistance of spores of non-proteolytic Cl. botulinum . These findings are relevant to the safety of minimally processed (e.g. sous-vide ) foods.     

深圳海域养殖牡蛎卫生质量状况(Ⅱ)——农药、微生物及麻痹性贝毒的含量与评价

为了了解深圳海域养殖牡蛎体内的卫生质量状况,我们于2002年1月～2004年2月对该海域养殖牡蛎体内的污染物水平进行了检测,结果显示六六六(BHC)为ND(未检出)～0.100mg·Kg^-1湿重,DDT为0.004～0.255mg·Kg^-1湿重,细菌总数为ND～9.3×10⁴Ind·g^-1,大肠菌群为ND～5.6×10⁴Ind·(100g)^-1,PSP为ND～34μg·(100g)^-1,副溶血性弧菌沙井和后海的近江牡蛎检测结果为阳性,其余站位未检出。参照GB18406.4-2001《农产品安全质量无公害水产品安全要求》,深圳海域养殖牡蛎的六六六、DDT、细菌总数和PSP完全符合要求,沙井和后海的近江牡蛎副溶血性弧菌超标,所有站位的大肠菌群均存在超标。综合重金属的检测结果发现,所有站位的牡蛎均有多个污染物超标,卫生质量欠佳,珠江口和深圳湾的近江牡蛎较大亚湾和大鹏湾的长牡蛎和褶牡蛎卫生质量更差。     

Bacterial Counts and Quality of Iced Fish Retailed at a Lusaka Market, Zambia

S ummary . The counts of total viable, coliform, streptococcal and sulphite reducing anaerobic bacteria and the presence of salmonellae were determined on 134 iced fish obtained from Luburma Market, Lusaka, Zambia, during June-December 1970. The quality of the uncooked fish was also assessed by appearance and odour. The purpose of these determinations was to obtain a picture of the variations of the bacterial counts in relation to season, origin, fish species and market quality. Total viable and coliform counts were of the order of millions and tens of thousands/cm² of skin surface, respectively. Higher counts were obtained in the hot season during September-October but with little change in appearance of the fish. There was a significant correlation ( P < 0·01) of both total viable and coliform bacteria with quality scores. A maximum permissible level of 10⁷ cells/cm² of skin surface was proposed for total viable counts and 10⁵/cm² for coliform bacteria, for iced fish of acceptable quality in Zambia.     

Plant regeneration from mesophyll protoplasts of Solanum commersonii dun

Somatic fusion of Solanum commersonii, a frost tolerant wild potato species not crossable with Solanum tuberosum, relies on the possibility to isolate and culture protoplasts. This study was conducted to determine whether protoplasts could be isolated and plants regenerated in three S. commersonii accessions. Shoot cultures for protoplast isolation were maintained on Murashige and Skoog medium. Mesophyll protoplasts were isolated and cultured using a protocol originally described for S. tuberosum with some modifications. Differences were evident among the three accessions for protoplast yield, plating efficiency and regeneration frequency. Protoplast yield ranged from 3.0 to 8.5 × 10⁶ protoplasts per g of fresh tissue. At 1–2 × 10⁴ protoplasts ml⁻¹, which was the optimal plating density, 10–20% of plated protoplasts gave multicellular colonies. Regeneration of shoots was observed in two accessions only, the maximum regeneration frequency being 66%. In one of these accessions the reduction of sucrose concentration in regeneration media improved the regeneration frequency from 14 to 35%. About three hundred plants were rooted in vitro and successfully transferred to soil.     

水产品和水体中灿烂弧菌现场可视化环介导等温扩增快检方法的建立及应用

基于环介导等温扩增(LAMP)技术建立水产品和养殖水域中灿烂弧菌现场可视化的快速、简便检测方法.以灿烂弧菌等作为研究对象,以灿烂弧菌的toxR基因作为靶基因,确定煮沸法为适合于弧菌基因组DNA提取的快捷方法,优化筛选的引物可以特异地检测灿烂弧菌,检测核酸浓度的灵敏度可以达到10-9g/L,并且结果稳定、可靠.采用该方法...     

A demonstration of bacterial conjugation within the alimentary canal of Rhabditis nematodes

Abstract: Rhabditis nematodes fed a diet of Escherichia coli defecate viable undigested bacteria. These bacteria retain phenotypic characteristics, including those encoded on plasmids. Nematodes can survive a 2-min surface sterilization with 2% chlorine bleach; internalized bacteria also survive this treatment and are released in the nematode wastes. Bacteria alone or on the surface of dead nematodes are unable to survive incubation with this solution. There were 3.2 × 10⁵ viable bacteria per nematode, indicating that sufficient bacteria were present for gene transfer. Transconjugants ( lac ⁻ nal ^R str ^R cm ^R) were recovered in the nematode fecal material following a protocol where nematodes were initially fed a plasmidless lac ⁻ nal ^R str ^S cm ^S E. coli and then, after surface sterilization, a lac ⁺ nal ^S E. coli plasmid donor containing the conjugative R100JA ( str ^R cm ^R) plasmid. The presence of plasmids in the transconjugants was confirmed by gel electrophoresis. The occurrence of conjugation in the gut was confirmed by dissection of individual surface-sterilized nematodes and isolation of transconjugants.     

新喀里多尼亚弧菌CGJ02-2中四氢嘧啶合成基因簇ectABC的克隆及其功能鉴定

研究旨在克隆新的四氢嘧啶合成基因簇,并对其功能进行鉴定,为应用于四氢嘧啶的生产奠定基础。从新喀里多尼亚弧菌CGJ02-2中克隆获得四氢嘧啶合成基因簇ectABC,ectABC与表达载体pBAD连接后转化至大肠杆菌BW25113中,通过L-阿拉伯糖诱导表达。采用SDS-PAGE和液质联用鉴定重组表达蛋白,利用全细胞催化合成四氢嘧啶,通过高分辨质谱鉴定四氢嘧啶,并从天冬氨酸浓度、KCl浓度、温度和pH 4个方面优化催化条件。结果表明,来自新喀里多尼亚弧菌CGJ02-2基因组的ectABC大小为2 235 bp。SDS-PAGE显示表达产物中有3个重组蛋白产生,液质联用鉴定表明其分子量分别与ectA、ectB、ectC的理论分子量一致。高分辨质谱分析发现全细胞催化上清中有四氢嘧啶产生。优化后的最适全细胞催化条件为：天冬氨酸浓度100 mmol·L^-1,KCl浓度100 mmol·L^-1,温度30℃,pH 7.0,最优条件下产量为1.11 mg·mL^-1。研究从弧菌中克隆了四氢嘧啶合成基因簇ectABC,并在大肠杆菌BW2511...