Bioremediation of oil polluted aquatic systems and soils with novel preparation `Rhoder'

This paper summarises the experience accumulated duringthe field application of biopreparation `Rhoder' (solely or in a combinationwith preliminary mechanical collection of free oil) for remediation of oil polluted aquatic systems and soils in the Moscow region and Western Siberia during 1994–1999.It was demonstrated that `Rhoder' had a very high efficiency (>99%) for bioremediation of the open aquatic surfaces (100 m² bay of the River Chernaya, two 5,000 m² lakes in Vyngayakha) at initial level of oil pollution of 0.4–19.1 g/l. During remediation of the wetland (2,000 m²) in Urai (initial level of oil pollution of 10.5 g/l), a preliminary mechanical collection of oil was applied (75% removal) followed by a triple treatment with `Rhoder'. It resulted in an overall treatment efficiency of 94%. Relatively inferior results of bioremediation of the 10,000 m² wetland in Vyngayakha (65% removal) and the 1,000 m² marshy peat soil in Nizhnevartovsk (19% removal) can be attributed to the very high initial level of oil pollution (24.3 g/l and >750 g/g dry matter, respectively) aggravated by the fact that it was impossible to apply a preliminary mechanical collection of oil on these sites. A possible strategy for remediation of such heavily polluted sitesis discussed.     

Enhancement in the viability and biosensing activity of freeze-dried recombinant bioluminescent bacteria

The genetically-engineeredEscherichia coli strain, DPD2540, which contains afabA::luxCDABE fusion gene, gives a bioluminescent output when membrane fatty acid synthesis is needed. For more practical application of this strain in the field as biosensor, freeze-drying was adopted. A 12% sucrose solution with Luria-Bertani (LB) broth, as determined by the viability after freeze-drying, was found to be the most effective composition for lyophilization solution among various compositions tested. Rapid freezing with liquid nitrogen also gave the best viability after freeze-drying as compared to samples frozen at −70°C and −20°C. The biosensing activities of the cells showed a greater sensitivity when the cells from the exponential phase were freeze-dried. Finally, the optimum temperature for use of the freeze-dried cells in the biosensor field was determined.     

Indicators of Microbial Sulfate Reduction in Acidic Sulfide-Rich Mine Tailings

Sulfate-reducing bacteria (SRB) are thought to be actively involved in the cycling of sulfur in acidic mine tailings. However, most studies have used circumstantial evidence to assess microbial sulfate activity in such environments. In order to fully ascertain the role of sulfate-reducing bacteria (SRB) in sulfur cycling in acidic mine tailings, we measured sulfate reduction rates, sulfur isotopic composition of reduced sulfide fractions, porewaters and solid-phase geochemistry and SRB populations in four different Cu-Zn tailings located in Timmins, Ontario, Canada. The tailings were sampled in the summer and in the spring, shortly after snowmelt. The results first indicate that all four sites showed very high sulfate reduction rates in the summer (～100–1000 nmol cm^{? 3}d^?1), which corresponded to the presence of sulfide in the porewaters and to high SRB populations. In some of the sites, zones of microbial sulfate reduction also corresponded to a decline of organic carbon and to an apparent pyrite (with slightly negative δ³⁴S values) enrichment around the same depth. Microbial sulfate reduction was also important in permanently acidic (pH 2–3) mine tailings sites, suggesting that SRB can be active under very acidic conditions. Secondly, the results showed that microbial sulfate reduction was greatly reduced in the spring, suggesting that temperature might be a key factor in the activity of SRB. However, a closer look at the results indicated that temperature was not the sole factor and that acidic conditions and limited substrate availability in the spring appeared to be important as well in limiting microbial sulfate par reduction in sulfidic mine tailings. Finally, the results indicate that sulfur undergoes rapid cycling throughout the year and that microbial sulfate reduction and metal sulfide precipitation do not appear to be a permanent sink for metals.     

杭州地区发酵乳制品中乳酸菌发生耐药传播的安全性研究

目的对杭州地区市售发酵乳制品中分离鉴定的乳酸菌进行耐药传播的安全性分析,旨在揭示发酵乳制品中是否存在可传递耐药基因的超级耐药菌。方法自行分离鉴定的耐药乳酸菌,提取质粒并电泳分析是否存在质粒;以耐受叠氮化钠的大肠埃希菌J53作为受体菌进行质粒接合试验;若乳酸菌存在耐药现象,发现存在耐药质粒,且耐药性可通过质粒传播,评定为3级。乳酸菌存在耐药现象,发现存在耐药质粒,但接合试验未能证明耐药性可通过质粒传播,评定为2级。乳酸菌存在耐药现象,但未检测出耐药质粒,评定为1级。结果美丽健产品中分离的双歧杆菌（MLJ-G-2）和乳杆菌（MLJ-G-1）检测出含有质粒,但不会发生耐药转移,三种菌耐药危害等级分别为2、2、1。伊利产品中分离的双歧杆菌（YL-G-3）、乳杆菌（YL-G-1）、嗜热链球菌（YL-Q）检测出含有质粒,但不会发生耐药转移,四种菌耐药危害等级分别为2、2、1、2。味全产品中分离的双歧杆菌（WQ-G-3）、乳杆菌（WQ-G-1）、乳杆菌（WQ-G-2）、嗜热链球菌（WQ-Q）均检测出含有质粒,其中乳杆菌（WQ-G-1）、乳杆菌（WQ-G-2）会发生耐药转移,四种菌耐药危害等级分别为2、3、3、2。光明产品中分离的乳杆菌（GM-G-2）、乳杆菌（GM-G-3）、嗜热链球菌（GMQ）检测出含有质粒,乳杆菌（GM-G-2）会发生耐药转移,四种菌耐药危害等级分别为2、1、3、2。蒙牛产品中只分离的嗜热链球菌（MN-Q）检测出含有质粒,不会发生耐药转移,耐药危害等级为2。结论发酵乳制品中分离所得的乳酸菌81.2%的菌能检测出质粒,耐药性可能与质粒相关,存在潜在的生物安全威胁;18.8%的菌的耐药性可通过质粒接合发生传播,存在直接的生物安全威胁。     

Sternal gland structures in males of bean flower thrips,Megalurothrips sjostedti,and Poinsettia thrips,Echinothrips americanus,in comparison with those of western flower thrips,Frankliniella occidentalis (Thysanoptera: Thripidae)

Sternal pores are important features for identification of male thrips, especially within the subfamily Thripinae. They vary in shape, size and distribution even between species of one genus. Their functional role is speculated to be that of sex- and/or aggregation pheromone production. Yet, sexual aggregations are not reported in Echinothrips americanus, known to have sternal pores, while we observed aggregations in Megalurothrips sjostedti, previously reported to lack them.We examined the sternal glands and pores of the thripine species E. americanus and M. sjostedti males, in comparison with those of Frankliniella occidentalis using light microscopy, as well as scanning and transmission electron microscopy. Pore plates of F. occidentalis were ellipsoid and medial on sternites III–VII, while in E. americanus they were distributed as multiple micro pore plates on sternites III–VIII. In M. sjostedti they appeared as an extremely small pore in front of the posterior margin of each of sternites IV–VII. Pore plate and pore plate area were distributed similarly on sternites III–VII in F. occidentalis. However, in E. americanus the total pore plate area increased significantly from sternites III to VIII. Ultrastructure of cells associated with sternal glands showed typical characteristics of gland cells that differ in size, shape and number. The function of sternal glands is further discussed on the basis of morphological comparisons with other thrips species.     

Bioavailability of Glucosyl Hesperidin in Rats

Glucosyl hesperidin (G-hesperidin) is a water-soluble derivative of hesperidin. We compared the absorption and metabolism of G-hesperidin with those of hesperidin in rats. After oral administration of G-hesperidin or hesperidin to rats, hesperetin was detected in sera hydrolyzed with β-glucuronidase, but it was not detectable in unhydrolyzed sera. Serum hesperetin was found more rapidly in rats administered G-hesperidin than in those administered hesperidin. The area under the concentration-time curve for hesperetin in the sera of rats administered G-hesperidin was approximately 3.7-fold greater than that of rats administered hesperidin. In the urine of both administration groups, hesperetin and its glucuronide were found. Urinary excretion of metabolites was higher in rats administered G-hesperidin than in those administered hesperidin. These results indicate that G-hesperidin presents the same metabolic profile as hesperidin. Moreover, it was concluded that G-hesperidin is absorbed more rapidly and efficiently than hesperidin, because of its high water solubility.     

烟草可培养内生细菌的分离及多样性分析

采用稀释平板法, 从健康烟草的根、茎、叶组织中分离到267株内生细菌。利用细菌菌落表征性状和16S rRNA序列对这些分离物进行了多样性分析。通过数值分析比较了8个菌落形态表征性状, 以类平均连锁聚类法的方式进行聚类分析, 在2.15的水平上可分成5个聚类群和56个亚群。16S rDNA序列系统发育分析表明267株分离物可分为21个类群。研究表明同一菌落形态类型的菌株在系统发育树中不一定聚为一类, 菌落形态分类与分子生物学方法分类结果不完全一致。经克隆测序分析表明, 这267株分离物分别与GenBank中6类细菌中的21个已知种相似性达到98%?99%。其中芽孢杆菌属(Bacillus)细菌是烟草可培养内生细菌的优势种群。     

Bacteria associated with orchid roots and microbial production of auxin

Associative bacteria of terrestrial (Paphiopedilum appletonianum) and epiphytic (Pholidota articulata) tropical orchids were investigated. Microbial community of epiphytic plant differed from that of the terrestrial one. Streptomyces, Bacillus, Pseudomonas, Burkholderia, Erwinia and Nocardia strains populated Paphiopedilum roots, whereas Pseudomonas, Flavobacterium, Stenotrophomonas, Pantoea, Chryseobacterium, Bacillus, Agrobacterium, Erwinia, Burkholderia and Paracoccus strains colonized Pholidota roots. Endophytic bacteria populations were represented with less diversity: Streptomyces, Bacillus, Erwinia and Pseudomonas genera were isolated from P. appletonianum, and Pseudomonas, Bacillus, and Flavobacterium genera were isolated from Ph. articulata. Microorganisms produced indole-3-acetic acid (IAA). Variations in its biosynthesis among the strains of the same genus were also observed. The highest auxin level was detected during the stationary growth phase. Biological activity of microbial IAA was proved by treatment of kidney bean cuttings with bacterial supernatants, revealing considerable stimulation of root formation and growth.