Starvation-induced barotolerance as a survival mechanism of a psychrophilic marine vibrio in the waters of the Antarctic convergence

Ant-300, a psychrophilic marine vibrio isolated from the Antarctic convergence, was tested for survival under conditions simulating those expected in situ for this organism. In particular, the organism's response to increased hydrostatic pressure was examined. Under a hydrostatic pressure of 250 atm, the viability of Ant-300 was reduced over 755 in growth media and under starvation conditions after 3 and 2 days, respectively. However, if the cells were starved for 1 week prior to pressurization of the starving suspension, 100% viability was maintained for over 6 weeks at the same pressure. After 10 weeks, the viability of a population of cells suspended for 1 week in natural or artificial seawater at a density of 10³ cells ml^–1 prior to pressurization of the suspension was 2 to 3 times greater than 1 atm controls. The data indicate that starvation conditions are a contributory if not the primary factor for the barotolerance of this organism in Antarctic waters. The data also indicate that under certain conditions, not unlike those expected in situ, hydrostatic pressure actually increases the survival of this organism under low nutrient conditions.Technical Paper No. 4907, Oregon Agricultural Experiment Station.     

Two different proteases produced by a deep-sea psychrotrophic bacterial strain,<Emphasis Type="Italic"> Pseudoaltermonas</Emphasis> sp. SM9913

A psychrotrophic bacterial strain, Pseudoaltermonas sp. SM9913, was isolated from deep-sea sediment collected at 1,855 m depth. Two proteases produced by Pseudoaltermonas sp. SM9913 were purified, MPC-01 and MCP-02. MCP-01 is a serine protease with a molecular weight of 60.7 kDa. It is cold-adapted with an optimum temperature of 30–35°C. Its K_m and E_a for the hydrolysis of casein were 0.18% and 39.1 kJ mol^–1, respectively. It had low thermostability, and its activity was reduced by 73% after incubation at 40°C for 10 min. MCP-02 is a mesophilic metalloprotease with a molecular weight of 36 kDa. Its optimum temperature for the hydrolysis of casein was 50–55°C. The K_m and E_a of MCP-02 for the hydrolysis of casein were 0.36% and 59.3 kJ mol^–1, respectively. MCP-02 had high thermostability, and its activity was reduced by only 30.5% after incubation at 60°C for 10 min. At low temperatures, Pseudoaltermonas sp. SM9913 mainly produced the psychrophilic protease MCP-01.Communicated by O. Kinne, Oldendorf/Luhe     

A comprehensive evaluation of the potential chemical defenses of antarctic ascidians against sympatric fouling microorganisms

The present study analyzed the bioactivity of whole body extracts from six solitary and eight colonial ascidian taxa against 20 sympatric bacterial isolates and one sympatric diatom species from the Western Antarctic Peninsula. Ascidians had crude lipophilic and hydrophilic extracts assayed against 20 bacterial strains. The lipophilic extract of one ascidian caused growth inhibition in all bacterial isolates at 3× tissue-level concentrations. The lipophilic and hydrophilic extracts were fractionated into seawater-soluble and insoluble fractions and assayed at three concentrations against a sympatric diatom species. Significant diatom mortality was detected at 3× and 1× concentrations in all but one ascidian taxon. Lipophilic fractions caused higher diatom mortality than hydrophilic extracts. The specificity of secondary metabolites against diatom fouling and the lack of activity against bacteria suggest high selective pressure for chemical defenses against diatom fouling or the potential that bacterial pathogens are controlled by the ascidian immune system.     

Bioactive potential of seagrass bacteria against human bacterial pathogens

Study of marine organisms for their bioactive potential, being an important part of marine ecosystem, has picked up the rhythm in recent years with the growing recognition of their importance in human life. Investigation was carried out to isolate 32 strains of endo and epiphytic bacteria in 2 seagrass species viz., Syringodium isoetifolium and Cymodocea serrulata. Morphologically different bacterial strains were tested against 5 antibiotic resistant human bacterial pathogens, of which 10 associated bacteria shown inhibitory activity against one or more bacterial pathogens. Minimum inhibitory concentration (MIC) and Minimum bacterial concentration (MBC) determination with extracellular bioactive compounds from the associated bacteria reveals that, the strain ENC 5 showed inhibitory activity against all the bacterial pathogens with the maximum sensitivity against Pseudomonas aeruginosa at the MIC value of 500 microg ml(-1).     

Activity of marine psychrophilic bacteria at elevated hydrostatic pressures and low temperatures

Uptake and metabolic turnover of a variety of radioactively labeled substrates by a number of bacterial isolates from several depths in offshore Atlantic waters were studied in laboratory experiments. The variable conditions were hydrostatic pressure (1 to 400 atm), temperature (1.50 to 15.0°C), concentration of substrate (2 to 500 g/ml), and time of incubation (4 h to 8 weeks). Elevated hydrostatic pressure and low temperature retarded the rate of metabolism (biosynthesis more than respiration) in all isolates tested, but considerably less in those of psychrophilic character.Contribution No. 2944 of the Woods Hole Oceanographic Institution.Supported by the National Foundation Grants GA-33405 and GA-29665.     

石灰性土壤中高效解磷细菌菌株的分离、筛选及组合

对石灰性土壤样品进行了解磷细菌的分离和筛选,得到具有明显溶磷圈的解磷细菌71株.分析了菌株分泌磷酸酶、释放速效磷及pH降低等综合因素和特征,研究了菌株生长动态.最后筛选出解磷能力较强且无拮抗反应的6株解磷细菌,并将其组合成为解磷细菌菌群.图8表2参17     

Temperature and pressure tolerance of larvae of Crepidula fornicata suggest thermal limitation of bathymetric range

The extant deep-sea fauna is thought to result from recolonisation of this environment by shallow-water organisms following climate-driven mass extinctions. Planktonic larval tolerance to high pressure is considered an important preadaptation for successful deep-sea invasion. In this study, the pressure and temperature tolerance of a species without any known confamilial deep-sea relative were assessed for the first time. Early- and late-veliger larvae of the shallow-water species Crepidula fornicata were subjected to a temperature/hydrostatic pressure regime from 5 to 25 °C and from 0.1 to 40 MPa. Although early and late veliger survived pressures equivalent to 2,000 m water depth or greater at all temperatures, decreased larval activity indicated significant sublethal temperature and pressure effects. Reduced larval activity of early veliger at low temperatures suggests that the bathymetric range of this species may be thermally constrained. A mechanistic model is proposed to explain the emerging pattern of ontogenetic shifts in pressure tolerance of shallow-water benthic invertebrates.     

Influence of hydrostatic pressure on l-glutamate dehydrogenase from the Antarctic fish Chaenocephalus aceratus

The influence of hydrostatic pressure on the stability and activity of glutamate dehydrogenase (GDH) from the liver of the Antarctic fish Chaenocephalus aceratus (Notothenioidei: Channichthyidae) has been investigated using the homologous bovine enzyme for comparison. At gradually increasing pressure, the fish GDH retains all activity up to 140 MPa, whereas in the same pressure range the bovine enzyme is slightly inactivated. The kinetics of pressure-induced inactivation of Antarctic GDH displays an exponentially decreasing residual activity as a function of the incubation time, whereas the bovine enzyme shows a complex pattern of behaviour in the first 20 min of incubation, attributable to the transient formation of aggregates. The coenzyme NAD increases the stability of both enzymes. The activation volume calculated for the reaction catalysed under pressure by fish GDH is higher than that of the bovine enzyme. These results indicate that the molecular properties of fish and bovine GDH are affected differently by hydrostatic pressure. A common mode of inactivation under extreme conditions of pressure and temperature of the cold-adapted Antarctic fish and mesophilic bovine GDH is deduced and discussed. Received: 20 October 1998 / Accepted: 7 May 1999     

Evidence of antibacterial and lysozyme-like activity in different planktonic larval stages of Paracentrotus lividus

Larval lysate (four-armed stage) exerts a lysozyme-like and antibacterial activity directed against Vibrio alginolyticus and other bacterial strains. This lysozyme activity is dependent on the pH and ionic strength of the reacting medium and sample. The other larval stages (six-armed, eight-armed and a stage approaching metamorphosis) possess protection against bacteria as well. The highest antibacterial activity is present in the six-armed stage. These antibacterial compounds avoid exploitation by bacteria. Paracentrotus lividus larvae used in our experiments were laboratory-reared from specimens collected in 1993 in Porto Cesareo (Lecce, Italy).     

产表面活性剂降解石油菌株产物性质及降解性能研究

微生物是组成生态系统的重要成员,在污染物去除中发挥着重要作用,是生物修复中的主力,然而在石油污染修复过程中,石油烃的疏水性会限制微生物对石油的降解,但一些微生物的细胞代谢物即生物表面活性剂,它是微生物在一定条件下代谢分泌产生的具有一定表面活性,集亲水基和疏水基结构于一分子的两亲性化合物,可以促进油的乳化,提高油的分散程度,增大菌株和油珠的接触机会,促进对石油烃的吸收和降解。在实验室分离得到了7株产表面活性剂石油降解菌株,经分子鉴定可知菌1和菌2都为粘质沙雷氏菌Serratia marcescens,菌3为居植物柔武氏菌Raoultella planticola,菌4,菌6和菌7都为克雷伯氏菌Klebsiella variicola,菌5为蜡状芽孢杆菌Bacillus cereus。主要研究了它们的生长与表面活性剂物质分泌状况的关系,发现随着时间增加,OD值随之增大,表面张力呈现下降趋势;并对菌株产物进行提取和薄层层析,离子型分析和红外光谱分析,初步判断其产物均为阴离子糖脂类;通过pH,初始油质量浓度,接种量和盐度4个单因素的变化研究菌1粘质沙雷氏菌,菌3居植物柔武士菌,菌5蜡状芽孢杆菌和菌6克雷伯氏菌对石油类物质降解能力,发现菌3居植物柔武氏菌和菌5蜡状芽孢杆菌降解性能较好;通过响应曲面法优化蜡状芽孢杆菌的降解条件,得出其最佳降解条件为pH为5.02,油质量浓度为3 g·L-1,接种量为1199.98μL,盐度为0.5 g·L-1时,在此条件下,菌株对石油的降解率为66.94%。     

Microbial interactions with the cyanobacterium Microcystis aeruginosa and their dependence on temperature

Associated heterotrophic bacteria alter the microenvironment of cyanobacteria and potentially influence cyanobacterial development. Therefore, we studied interactions of the unicellular freshwater cyanobacterium Microcystis aeruginosa with heterotrophic bacteria. The associated bacterial community was greatly driven by temperature as seen by DNA fingerprinting. However, the associated microbes also closely interacted with the cyanobacteria indicating changing ecological consequence of the associated bacterial community with temperature. Whereas concentration of dissolved organic carbon in cyanobacterial cultures changed in a temperature-dependent manner, its quality greatly varied under the same environmental conditions, but with different associated bacterial communities. Furthermore, temperature affected quantity and quality of cell-bound microcystins, whereby interactions between M. aeruginosa and their associated community often masked this temperature effect. Both macro- and microenvironment of active cyanobacterial strains were characterized by high pH and oxygen values creating a unique habitat that potentially affects microbial diversity and function. For example, archaea including ‘anaerobic’ methanogens contributed to the associated microbial community. This implies so far uncharacterized interactions between Microcystis aeruginosa and its associated prokaryotic community, which has unknown ecological consequences in a climatically changing world.     

Larval settlement in the serpulid polychaete Hydroides elegans in response to bacterial films: an investigation of the nature of putative larval settlement cue

Larval settlement in the marine polychaete Hydroides elegans (Haswell) is induced by certain bacteria in marine biofilms. The exact nature of the settlement cue that larvae of H. elegans receive from these bacteria remains unknown. In this study, we revealed some properties of the bacterially derived larval settlement cue by investigating the larval settlement inductive activity of two bacterial strains after various treatments. These two bacterial strains, Roseobacter sp. and an α-subclass Proteobacteria, are highly inductive to larval settlement of H. elegans. The larvae responded similarly to Roseobacter and Proteobacteria in all the larval settlement bioassays, suggesting that the larval settlement-inducing substances produced by these bacteria may share common characteristics. First of all, the larvae did not settle in the seawater conditioned by the bacteria attached as a film or by the bacteria that were freely suspended in seawater. The results suggest that the putative larval settlement cue is not released into seawater and, therefore, should be associated with the surface of the bacteria. Secondly, formaldehyde treatment entirely eliminated the larval settlement induction activity of the bacterial films, and streptomycin treatment reduced the percentage of larval settlement on the bacterial films in a concentration-dependent manner. Since both treatments can kill bacteria with little damage to the surface chemistry of bacterial cells, the decline in larval settlement is suggested be due to a reduction of the viable bacterial population in the bacterial films. In fact, the reduction of larval settlement in the streptomycin treatments coincided with the decrease in viable bacterial populations in broth cultures containing respective concentrations of streptomycin. These results suggest that the viability of Roseobacter and Proteobacteria is important to their settlement induction effect. Since the larval settlement induction activity of the bacterial strains appears to correlate with their viability, we suggest that the putative larval settlement cue is derived from a metabolic pathway in the bacteria and that the cue is exported to and concentrated at the extracellular polymer matrix of the bacterial cell, at which the larvae establish contact with the bacteria. The larval settlement cue may be highly susceptible to degradation so that a metabolically active bacterial film is needed to maintain the putative cue at a concentration that surpasses the threshold for induction of larval settlement. Received: 14 October 1998 / Accepted: 5 September 2000     

Antimicrobial activity of extracts of Caribbean gorgonian corals

Extracts of 39 species of Caribbean gorgonians were tested for antimicrobial activity against 15 strains of marine bacteria. The bacteria consisted of three opportunistic pathogens, Vibrio parahaemolyticus, Leucothrix mucor, and Aerococcus viridans, and 12 strains isolated from either healthy or decayed gorgonians. Overall, only 15% (79 out of 544) of the tests resulted in antibacterial activity with 33% (13 out of 39) of the gorgonians inhibiting only one bacterial strain and 23% (9 out of 39) showing no activity. The extracts of four Pseudopterogorgia species showed relatively high levels of activity, inhibiting 43 to 86% of the bacterial strains. The potency of the active Pseudopterogorgia species was variable, however, and three additional Pseudopterogorgia species were inactive against all bacterial strains. With the exception of one sensitive strain, Vibrio species were resistant to gorgonian metabolites. Our results indicate that organic extracts of most Caribbean gorgonians do not possess potent, broad-spectrum antibacterial activity inhibitory to the growth of opportunistic marine pathogens and bacteria associated with healthy and decayed gorgonian surfaces. These findings suggest that the inhibition of bacterial growth is not the primary ecological function of gorgonian secondary metabolites and that bacteria may not be important selective agents in the evolution of gorgonian secondary chemistry.     

Alleviation of nickel stress by halophilic bacteria in mungbean(Vigna radiata)

Mungbean is an important, short-duration legume crop with high nutritive value for poor families, where protein and micronutrient scarcity are most pervasive. In developing countries, like Pakistan, where mungbean is one of the greatest sources of protein for underprivileged people, salinity and metal stresses are drastically limiting its yield. Therefore, it is necessary to explore the ways for promoting the growth of mungbean under stressed conditions. This study provides the basis for the use of two previously isolated halophilic and nickel-resistant bacteria [Bacillus subtilis and Halomonas aquamarina(1)] in promoting the growth of mungbean plant under nickel-stressed environments. These bacterial strains were previously found to improve the growth of plants under salinity because of their good biofilm-forming abilities. We hypothesized that these two halophilic and nickel-resistant biofilm-forming bacteria can help the plants to grow under nickel-stressed environments by promoting soil aggregation as they were previously reported to resist salinity and nickel stress, as well as were found to promote plant growth under saline environments. The results revealed that under nickel stress, both bacteria promote plant growth by facilitating the formation of soil aggregates through an increase in their extra polymeric substance(EPS) production.     

The detrital food chain based on seaweeds. I. Bacteria associated with the surface of Laminaria fronds

The bacteria associated with the surface of fronds of the sublittoral brown alga Laminaria longicruris were investigated over a 13-month period on the coast of Nova Scotia (Canada). A psychrophilic population was found to be associated with the frond during the winter and a mesophilic population with the decaying frond during the summer. Numbers of psychrophiles varied inversely with ambient water temperature, and were present in the greatest number on the eroded tips of fronds. Laminaran hydrolyzing isolates were characteristic of the psychrophilic flora and a group of isolates hydrolyzing mannitol, protein and alginate characteristic of the mesophilic population. Increases in the numbers and proportions of bacteria utilizing plant substrates were found to accompany macroscopic evidence of frond decomposition. A comparison of the bacterial floras of L. longicruris fronds from a sheltered and exposed location showed them to be quantitatively and qualitatively comparable.     

Evidence of high organotrophic potentiality of bacteria in the deep ocean

The organotrophic potentiality of heterotrophic microorganisms 50 cm above the ocean floor (5,207 m depth) was measured in situ, at a station in the subarctic Pacific Ocean (44° N; 154° E). In certain enriched deep-sea samples incubated in situ, the microorganisms exhibited nearly the same organotrophic activity as those in the surface water layers. Rod-shaped barophilic bacteria (0.4 to 0.6 μ x 1.0 to 2.0 μ), which multiplied at a generation time of about 10 h, were responsible for this activity.     

Biodegradation of phenol by native microorganisms isolated from coke processing wastewater

The present investigation was undertaken to assess the biodegradation of phenol by native bacteria strains isolated from coke oven processing wastewater. The strains were designated ESDSPB1, ESDSPB2 and ESDSPB3 and examined for colony morphology Gram stain characters and biochemical tests. Phenol degrading performance of all the strains was evaluated initially. One of the strains namely ESDSPB2 was found to be highly effective for the removal of phenol, which was used as sole carbon and energy source. From an initial concentration of 200 mg I(-1) it degraded to 79.84 +/- 1.23 mg l(-1). In turn the effect of temperature (20 to 45 degrees C), pH (5-10) and glucose concentration (0, 0.25 and 0.5%) on the rate of phenol degradation by that particular strain was investigated. Observations revealed that the rate of phenol biodegradation was significantly affected by pH, temperature of incubation and glucose concentration. The optimal conditions for phenol removal were found to be pH of 7 (84.63% removal), temperature, 30 degrees C (76.69% removal) and 0.25% supplemented glucose level (97.88% removal). The main significance of the study is the utilization of native bacterial strains from the waste water itself having potential of bioremediation.     

秸秆污泥堆肥产纤维素酶细菌的筛选及产酶条件优化

从添加秸秆进行堆肥处理的污泥中采集样品,通过富集培养和刚果红平板染色法筛选分离出具有纤维素降解能力的细菌,再通过酶活力测定从中分离筛选出1株相对高活性的产纤维素酶细菌CI;通过基于16S rRNA基因序列的系统发育分析,初步确定该菌株为Devosia sp..利用单因素试验对目的菌株C1进行产纤维素酶发酵条件优化,结果表明菌株C1产纤维素酶的最佳发酵时间、培养温度、摇床转速以及最适初始pH值分别为60 h、30℃、130 r·min^-1和7.2～7.5,且在该条件下其滤纸酶（FPase）和羧甲基纤维素酶（CMCase）活力分别达23.10和54.97 U·mL^-1.     

Larval viability and heterozygote deficiency in populations of marine bivalves: evidence from pair matings of mussels

Using a pressurized temperature gradient, which rapidly equilibrates bacteria to a wide range of temperature and pressures, temperature ranges for growth and survival of Vibrio harveyi (strain B 392) were found to narrow with increasing pressures. Both temperature and pressure ranges were greater in an enriched seawater medium than in unenriched seawater. Using a kinetics apparatus, V. harveyi was also exposed to gradual changes in temperature and pressure, simulating a particle sinking at approximately 200 m d^-1 from surface waters (17 °C; 1 bar pressure) to 3 400 m (4°C; 340 bars pressure). With gradual temperature changes, cells were able to grow and survive at lower temperatures than they had in rapid equilibration experiments. Gradual pressurization, however, did not increase pressure tolerance. Under all experimental conditions, the combined effects of low temperatures and high pressures were lethal for V. harveyi. It is hypothesized that temperature and pressure stratify the ocean into regions of growth, survival, and death for V. harveyi, and perhaps other bacteria.     

The effect of pressure on muscle lactate dehydrogenase activity of some deep-sea and shallow-water fishes

The activity of crude muscle lactate dehydrogenase (LDH) of several species of bathypelagic and shallow-water fishes has been measured at pressures between 1 and 578 atm and at temperatures of 15° and 25°C. No relationship has been found between the effect of pressure on enzyme activity and the hydrostatic pressure of the organism's environment. Applied hydrostatic pressure reduced activity at both temperatures. The decrease at 25°C was double the decrease at 15°C in LDH from shallow-water fishes. However, enzymes from 2 bathypelagic fishes showed approximately the same reduction at both temperatures. Thus, the interaction of temperature and pressure was less in deep-sea than in shallow-water fish LDH. Decreasing temperature and increasing pressure would both reduce the activity of LDH. That is, deep-sea conditions are noncompensatory in this instance. It is possible that the dissociation of the effects of temperature and pressure could be an adaptive feature of deep-sea life.This paper is a portion of a thesis submitted to the Graduate School, University of Georgia, in partial fulfillment for the degree of Master of Science.