Induction of maleate hydratase in Pseudomonas pseudoalcaligenes

Maleate hydratase (malease, EC 4.2.1.31) activity in P. pseudoalcaligenes is induced when grown on3-hydroxybenzoate. The specific malease activity was constant during the logarithmic phase in a batch culture containing 3-hydroxybenzoate as the carbon source, when 3-hydroxybenzoate-grown cells were used as inoculum. When yeast extract-grown cells were used as inoculum, the specific malease activity was correlated with growth. In both instances the specific malease activity dropped rapidly as soon as growth ceased. Maleate did not serve as a growth substrate for this microorganism, but a mutant able to grow on maleate was selected. The specific malease activity of maleate-grown cells of this mutant was not higher than the basal level of induction of malease activity.     

Pseudomonas pseudoalcaligenes B50铁载体合成相关基因pyrD的克隆与功能分析

从棉花根际分离的一株细菌B50在低铁条件下可以产生铁载体。通过形态学特征、生理生化特征及其16SrDNA序列分析,将其鉴定为Pseudomonas pseudoalcaligenes。采用三亲本杂交的方法将转座子Tn5-1063a(含luxAB)导入B50中,进行转座子插入诱变,获得突变株。用TAIL-PCR方法得到与铁吸收有关的pyrD基因序列。通过互补实验验证pyrD与铁载体的合成有关。Pseudomonas pseudoalcaligenes能够产生铁载体属于首次报道。     

Control of the pyrimidine biosynthetic pathway in Pseudomonas pseudoalcaligenes

The five de novo enzyme activities unique to the pyrimidine biosynthetic pathway were found to be present in Pseudomonas pseudoalcaligenes ATCC 17440. A mutant strain with 31-fold reduced orotate phosphoribosyltransferase (encoded by pyrE) activity was isolated that exhibited a pyrimidine requirement for uracil or cytosine. Uptake of the nucleosides uridine or cytidine by wild-type or mutant cells was not detectable; explaining the inability of the mutant strain to utilize either nucleoside to satisfy its pyrimidine requirement. When the wildtype strain was grown in the presence of uracil, the activities of the five de novo enzymes were depressed. Pyrimidine limitation of the mutant strain led to the increase in aspartate transcarbamoylase and dihydroorotate dehydrogenase activities by more than 3-fold, and dihydroorotase and orotidine 5-monophosphate decarboxylase activities about 1.5-fold, as compared to growth with excess uracil. It appeared that the syntheses of the de novo enzymes were regulated by pyrimidines. In vitro regulation of aspartate transcarbamoylase activity in P. pseudoalcaligenes ATCC 17440 was investigated using saturating substrate concentrations; transcarbamoylase activity was inhibited by P_i, PP_i, uridine ribonucleotides, ADP, ATP, GDP, GTP, CDP, and CTP.     

Effect of temperature on growth parameters of psychrophilic bacteria and yeasts

Three bacterial (Pedobacter heparinus, Pedobacter piscium, Pedobacter cryoconitis) and three yeast strains (Saccharomyces cerevisiae, Leucosporidiella creatinivora, Rhodotorula glacialis) of different thermal classes (mesophiles and psychrophiles) were tested for the effect of temperature on a range of growth parameters, including optical density, viable cell numbers, and cell dry mass, in order to determine the temperature conditions under which maximum biomass formation is obtained. Maximum values of growth parameters obtained at the stationary growth phase of the strains were used for statistical calculation. Temperature had a significant (P ≤ 0.05) effect on all growth parameters for each strain; correlations between the growth parameters were significant (P ≤ 0.05–0.01). The maximum growth temperature or the temperature at which microbial growth was fastest was in no case the temperature at which the investigated strains produced the highest amount of biomass. All tested psychrophilic bacteria and yeast strains produced highest amounts of cells (as calculated per mg cell dry mass or per OD₆₀₀ unit) at 1°C, while cell numbers of mesophiles were highest at 20°C. Thus, cultivation temperatures close to the maximum growth temperature are not appropriate for studying psychrophiles.     

Domesticated and wild fathead minnows differ in growth and thermal tolerance

Many populations have evolved in response to laboratory environments (lack of predators, continual food availability, etc.). Another potential agent of selection in the lab is exposure to constant thermal environments. Here, we examined changes in growth, critical thermal maximum (CT_max), and food consumption under constant (25 °C) and fluctuating (22–28 °C and 19–31 °C) conditions in two populations of fathead minnows, Pimephales promelas: one that has been kept in a laboratory setting for over 120 generations (~40 years) and a corresponding wild one. We found that under thermal fluctuations, domesticated fathead minnows grew faster than their wild counterparts, but also exhibited lower thermal tolerance. Food consumption was significantly higher in the lab population under the constant and large fluctuation thermal treatments. Our results suggest that the lab population has adjusted to the stable conditions in the laboratory and that we should carefully apply lessons learned in the lab to wild populations.     

The effects of temperature on bacterial production in a dimictic eutrophic lake

Abstract The incorporation of [³H](methyl)thymidine into DNA by the planktonic heterotropic bacteria of Little Crooked Lake (Noble Country, IN) was determined at different incubation temperatures. The highest rates of thymidine incorporation generally occurred at temperatures exceeding the in situ temperature of the sample. The optimal temperature for thymidine incorporation ranged from 1.0–3.4 times the in situ temperature. As the summer of 1983 progressed, the optimal temperatures for thymidine incorporation by epilimnetic samples and the in situ temperatures converged. This trend was reversed as fall overturn was approached.     

Mutational neighbourhood and mutation supply rate constrain adaptation in Pseudomonas aeruginosa

Understanding adaptation by natural selection requires understanding the genetic factors that determine which beneficial mutations are available for selection. Here, using experimental evolution of rifampicin-resistant Pseudomonas aeruginosa, we show that different genotypes vary in their capacity for adaptation to the cost of antibiotic resistance. We then use sequence data to show that the beneficial mutations associated with fitness recovery were specific to particular genetic backgrounds, suggesting that genotypes had access to different sets of beneficial mutations. When we manipulated the supply rate of beneficial mutations, by altering effective population size during evolution, we found that it constrained adaptation in some selection lines by restricting access to rare beneficial mutations, but that the effect varied among the genotypes in our experiment. These results suggest that mutational neighbourhood varies even among genotypes that differ by a single amino acid change, and this determines their capacity for adaptation as well as the influence of population biology processes that alter mutation supply rate.     

北京地区实验动物绿脓杆菌分离株MLVA分型

目的通过多位点可变数目串联重复序列分析（multiple-locus variable-number tandem-repeat analysis,MLVA）分型方法,研究北京地区实验动物绿脓杆菌分离株基因型和分布情况。方法选择13个可变数目重复序列（variable-number tandem-repeat,VNTR）位点,对实验动物及设施中检测出的141株绿脓杆菌的基因组DNA进行重复序列扩增,所得指纹图谱使用BioNumerics软件进行聚类分析,绘制系统发育树和最小生成树（minimum spanning tree,MST）。结果所采用的13个VNTR位点能够对全部分离株进行有效分型。141株绿脓杆菌主要被分为了3个基因群,56个基因型。各群所占比例分别为A群82.3%,B群占12.8%,C群占5.0%,辛普森多样性指数为0.763。同一区域内相邻实验动物单位的绿脓杆菌分离株同源关系较远。结论 MLVA方法对绿脓杆菌具有很好的分型能力,能够有效的追踪绿脓杆菌的来源。北京地区实验动物中绿脓杆菌分离株基因型多态性丰富,但无地域性同源关系。     

Effect of rearing temperature on growth and thermal tolerance of Schizothorax (Racoma) kozlovi larvae and juveniles

Effect of rearing temperature on growth and thermal tolerance of Schizothorax (Racoma) kozlovi Nikolsky larvae and juveniles was investigated. The fish (start at 12 d post hatch) were reared for nearly 6 months at five constant temperatures of 10, 14, 18, 22 and 26 °C. Then juvenile fish being acclimated at three temperatures of 14, 18 and 22 °C were chosen to determine their critical thermal maximum (CTMax) and lethal thermal maximum (LTMax) by using the dynamic method. Growth rate of S. kozlovi larvae and juveniles was significantly influenced by temperature and fish size, exhibiting an increase with increased rearing temperature, but a decline with increased fish size. A significant ontogenetic variation in the optimal temperatures for maximum growth were estimated to be 24.7 °C and 20.6 °C for larvae and juveniles of S. kozlovi, respectively. The results also demonstrated that acclimation temperature had marked effects on their CTMax and LTMax, which ranged from 32.86 °C to 34.54 °C and from 33.79 °C to 34.80 °C, respectively. It is suggested that rearing temperature must never rise above 32 °C for its successful aquaculture. Significant temperature effects on the growth rate and thermal tolerance both exhibit a plasticity pattern. Determination of critical heat tolerance and optima temperature for maximum growth of S. kozlovi is of ecological significance in the conservation and aquaculture of this species.     

波动温度下罗非鱼特定腐败菌生长动力学模型和货架期预测

研究了0℃~15℃范围的波动温度条件下,有氧贮藏养殖罗非鱼的特定腐败菌-假单胞菌的生长动力学模型及其对剩余货架期预测的适用性。由Belehradek方程建立了温度对假单胞菌生长动力学影响的数学模型,在设计的两种波动温度条件下假单胞菌生长动力学模型的预测值,与波动温度贮藏罗非鱼中假单胞菌生长的实测值比较,偏差度在0.906~0.942之间,准确度在1.13~1.19之间。以假单胞菌生长动力学模型预测的剩余货架期,与波动温度贮藏罗非鱼的感官、VBN和假单胞菌数评价获得的实测剩余货架期相比较,相对误差分别为5.9%和-9.1%。显示在贮藏温度波动的情况下,假单胞菌生长动力学模型同样可以快速可靠地实时预测0℃~15℃贮藏的罗非鱼剩余货架期。     

The distribution of fitness effects of new beneficial mutations in Pseudomonas fluorescens

Theoretical studies of adaptation emphasize the importance of understanding the distribution of fitness effects (DFE) of new mutations. We report the isolation of 100 adaptive mutants—without the biasing influence of natural selection—from an ancestral genotype whose fitness in the niche occupied by the derived type is extremely low. The fitness of each derived genotype was determined relative to a single reference type and the fitness effects found to conform to a normal distribution. When fitness was measured in a different environment, the rank order changed, but not the shape of the distribution. We argue that, even with detailed knowledge of the genetic architecture underpinning the adaptive types (as is the case here), the DFEs remain unpredictable, and we discuss the possibility that general explanations for the shape of the DFE might not be possible in the absence of organism-specific biological details.     

Body temperature of ostriches (Struthio camelus) kept in an open stable during winter time in Germany

Commercial breeding of ostriches in the northern parts of Europe is still discussed controversially in particular in winter. Here we measured the body temperature of 12 ostriches of two different ages during 12 weeks in winter using temperature loggers implanted subcutaneously and in the abdominal cavity. The mean peritoneal temperature ranged between 37.4 and 38.5 °C in the younger subjects and between 37.2 and 38.1 °C in the older subjects. Climatic conditions significantly affected the body temperature but in comparison with the variations between and within subjects these effects were low. There was no evidence that the young ostriches were not able to adapt towards the climatic conditions prevalent during our study.     

Effect of temperature on diauxic growth with glucose and organic acids in Pseudomonas fluorescens

Growth of Pseudomonas fluorescens in batch culture with glucose and organic acids resulted in typical diauxic responses at 30° C but no detectable diauxic lag at 5° C.At 30° C, organic acids were preferentially utilized during the first growth phase. Glucose utilization was delayed unitl onset of the second growth phase. Systems involved in direct uptake and catabolism of glucose responded in a manner compatible with respression by malate and/or its metabolites and induction by glucose and/or its metabolites. The oxidative non-phosphorylated pathway, through gluconate and 2-ketogluconate (2-KG) as intermediates, was not induced during either growth phase.At 5° C, growth with glucose and organic acids was biphasic but without diauxic lag. Organic acids were preferentially utilized during the first growth phase. Although carbon from glucose was not fully catabolized until onset of the second growth phase, glucose was oxidized to and accumulated extracellularly as gluconate and 2-KG during the first growth phase. No significant repression of glucose-catabolizing enzymes was observed during growth with organic acids in the presence of glucose. However, uptake activities for gluconate and 2-KG did not increase significantly until onset of the second growth phase.Thus, at low temperatures, psychrotrophic P. fluorescens oxidized glucose to extracellular 2-KG, while growing on preferred carbon sources. The 2-KG was then catabolized after depletion of the organic acid.     

Mutation of rpoS enhances Pseudomonas sp. KL28 growth at higher concentrations of m-cresol and changes its surface-related phenotypes

A Tn5 transposon mutant was isolated of the alkylphenol degrader Pseudomonas sp. KL28 that showed increased growth at higher levels of m-cresol on solid and in liquid cultures. The transposon was inserted at the 5'-terminus of rpoS, which encodes a stationary-phase sigma factor. When grown on agar plates, the rpoS mutant developed prominent wrinkles, especially at lower temperatures, and spread faster on soft agar. In addition, the rpoS mutant had enhanced biofilm-forming ability that was not due to self-produced diffusible signals.     

Fluorescent Pseudomonas mainly produce the dihydro form of pyoverdine at low specific growth rate

AIMS: To analyse the influence of cell growth rate and iron concentration on the production of pyoverdines (PVDs) and of their reduced dihydro forms by three fluorescent Pseudomonas strains (P. putida BTP16, P. fluorescens BTP7 and P. aeruginosa 7NSK2). METHODS: PVD and dihydropyoverdine (DHPVD) productions were determined by LC ESI-MS and spectrophotometry during batch and chemostat culture at different dilution rates. SIGNIFICANCE: The relatively high PVD-to-DHPVD ratio (0.57) observed in pH-controlled batch cultures suggested that a base-catalysed chemical oxidation of the dihydroform is not the prime mechanism involved in generating PVDs. Interestingly, in chemostat cultures the PVD-to-DHPVD ratio was significantly reduced at low specific growth rate. Our results suggest that the oxidation of DHPVD to PVD is catalysed by an iron-dependent enzymatic reaction rather than a chemical oxidation.     

肺部铜绿假单胞菌感染前后肺组织角质细胞生长因子的变化研究

目的观察与分析肺部铜绿假单胞菌（PAE）感染前后患者肺组织角质细胞生长因子（KGF）的变化情况,并研究其临床意义。方法将2010年12月到2013年12月于沈阳军区总医院接受治疗的60例肺炎患者作为研究对象,分别于其感染前后收集肺组织标本,检测感染前后患者肺组织角质细胞生长因子的表达水平。结果在感染PAE后,患者肺组织角质细胞生长因子蛋白表达水平呈明显上升的态势,并于感染3 d后到达峰值。相较感染铜绿假单胞菌前,患者肺组织角质细胞生长因子蛋白表达水平明显提升,感染前后对比差异有统计学意义（P〈0.05）。结论在急性肺损伤的诊断与治疗中,重视对患者肺组织KGF蛋白表达水平的研究,及时实施KGF辅助干预,对抑制感染,控制肺炎的进展有十分重要的作用。