Microbiological studies of Tokyo Bay

The generic composition of the heterotrophic bacterial population of Tokyo Bay, which is now highly polluted and eutrophic, was compared with that of the adjacent, less polluted regions of Sagami Bay and Suruga Bay. Members of Vibrionaceae predominated in the bacterial flora of seawater and zooplankton samples from Sagami Bay, Suruga Bay, and the mouth of Tokyo Bay. However,Vibrio spp. formed only a small proportion of the bacterial population of the water and sediment samples from the inner Tokyo Bay; there the Gram-negative, nonmotile, nonpigmented bacteria, which were tentatively identified asAcinetobacter, were predominant. The result of experiments, in which seawater samples from Tokyo Bay were incubated under various experimental conditions, indicated that two significant factors apparently control the growth ofVibrio spp. in seawater; (1) a direct antagonism betweenVibrios and phytoplankton undergoing rapid growth, and (2) a limiting organic nutrient forvibrios.     

Denitrification, Nitrate Reduction, and Oxygen Consumption in Coastal and Estuarine Sediments

Denitrification and consumption of oxygen and nitrate in sediments from Tama Estuary, Odawa Bay, and Tokyo Bay were measured in an experimental sediment-water system. Filtered seawater containing [¹⁵N]nitrate flowed continuously over undisturbed sediments, and the concentrations of O₂, nitrate, and nitrite in the influent and effluent and of ¹⁵N₂ in the effluent were monitored. Under steady-state conditions, the rate of nitrate consumption was the same order of magnitude as the rate of oxygen consumption in Tama Estuary sediments, whereas the former rate was one order of magnitude lower than the latter rate in Odawa Bay and Tokyo Bay sediments. Denitrification accounted for 27 to 57% of the nitrate consumption.     

Fluctuation of deoxyribonuclease activity from late spring to autumn in Tokyo Bay

Deoxyribonuclease (DNase) activity in seawater of Tokyo Bay was determined during 1972 and 1973. The activity was dependent on substrate (deoxyribonucleic acid) concentration within the range from o to io mg/l. During the summer the activity was high in the inner part of the bay and decreased on approaching the mouth of the bay. The intensity of activity was very high compared with the standing crop of particulate deoxyribonucleic acid (DNA) and this high activity corresponded with rapid decrease of the amount of nucleic acid with increasing depth in the water columns in the bay.During the investigation carried out in 1972 the activity fluctuated, being weak in May and November and high during the summer months. The profiles for activity did not coincide with those for chlorophyll-a, particulate DNA and seston, except in September. These non-parallel relations are discussed in terms of the distribution patterns of phytoplankton and bacteria.     

Aerobic cellulolytic bacterial flora associated with decomposing Phragmites leaf litter in a seawater lake

A litter bag experiment was carried out in a eutrophic seawater lake from autumn to summer in order to determine which bacterial genera play an important role in decomposition of Phragmites communis leaf litter. The count of cellulolytic bacteria and decomposition rate of litter cellulose increased rapidly during the initial month. In contrast, the count of cellulolytic fungi was lowest in this period. Pseudomonas accounted for 65–90% of total isolates of cellulolytic bacteria up to 5 months. These results suggest that Pseudomonas plays an important role in at least the initial decomposition stage of the litter.     

Size and species-specific primary productivity and community structure of phytoplankton in Tokyo Bay

Combined methods of size fractionation and single-cell isolationwere used to investigate the seasonal variation of phytoplanktondynamics in Tokyo Bay with an emphasis on primary productivity.Red tides occurred in Tokyo Bay from spring to autumn; a diatom,Skeletonema costatum, and a raphidophycean, Heterosigma akashiwo,were the most important primary producers. Small diatoms andflagellates, including these species, were dominant and showedrapid changes of phytoplankton community structure within severaldays in summer. The nanoplankton (3–20 µm) fractioncontributed most to chlorophyll a concentration and primaryproductivity during spring to autumn, whereas the microplankton(>20 µm) contribution was remarkable in winter. Picoplankton(<3 µm phytoplankton) remained relatively constantthroughout the year. A significant reverse relationship wasobtained between assimilation rate and chlorophyll a contentfor the total and nanoplankton population; the assimilationrate was high at the initial phase of the bloom, then decreasedto a minimum level at the peak of the bloom. Factors controllingthe reduction of assimilation rates at the peak, and changesin phytoplankton community structure, are discussed.     

Environmental factors influencing the rate of hydrocarbon oxidation in temperate lakes.

Rates of hydrocarbon biodegradation were estimated by following oxygen uptake during mineral oil oxidation or oxidation of [1-14C]hexadecane to 14CO2, when these substrates were added to natural water samples from Wisconsin lakes. A lag phase preceded hydrocarbon oxidation, the length of which depended on population density or on factors influencing growth rate and on the presence of nonhydrocarbon organic compounds. Hydrocarbon oxidation was coincident with growth and presumably represented the development of indigenous hydrocarbon-degrading microorganisms in response to hydrocarbon additions. In detailed studies in Lake Mendota, it was found that, despite the continued presence of hydrocarbon-degrading microorganisms in water samples, seasonal variations in the rates of mineral oil and hexadecane oxidation occurred which correlated with seasonal changes in temperature and dissolved inorganic nitrogen and phosphorus. The temperature optimum for oil biodegradation remained at 20 to 25 C throughout the year, so that temperature was the main limiting factor during winter, spring, and fall. During summer, when temperatures were optimal, nutrient deficiencies limited oil biodegradation, and higher rates could be obtained by addition of nitrogen and phosphorus. The rates of hydrocarbon biodegradation were thus high only for about 1 month of the ice-free period, when temperature and nutrient supply were optimal. Nutrient limitation of oil biodegradation was also demonstrated in 25 nutrient-poor lakes of northern Wisconsin, although in almost every case oil-degrading bacteria were detected. Knowledge of temperature and nutrient limitations thus will help in predicting the fate of hydrocarbon pollutants in freshwater.     

Environmental factors influencing the rate of hydrocarbon oxidation in temperate lakes.

Rates of hydrocarbon biodegradation were estimated by following oxygen uptake during mineral oil oxidation or oxidation of [1-14C]hexadecane to 14CO2, when these substrates were added to natural water samples from Wisconsin lakes. A lag phase preceded hydrocarbon oxidation, the length of which depended on population density or on factors influencing growth rate and on the presence of nonhydrocarbon organic compounds. Hydrocarbon oxidation was coincident with growth and presumably represented the development of indigenous hydrocarbon-degrading microorganisms in response to hydrocarbon additions. In detailed studies in Lake Mendota, it was found that, despite the continued presence of hydrocarbon-degrading microorganisms in water samples, seasonal variations in the rates of mineral oil and hexadecane oxidation occurred which correlated with seasonal changes in temperature and dissolved inorganic nitrogen and phosphorus. The temperature optimum for oil biodegradation remained at 20 to 25 C throughout the year, so that temperature was the main limiting factor during winter, spring, and fall. During summer, when temperatures were optimal, nutrient deficiencies limited oil biodegradation, and higher rates could be obtained by addition of nitrogen and phosphorus. The rates of hydrocarbon biodegradation were thus high only for about 1 month of the ice-free period, when temperature and nutrient supply were optimal. Nutrient limitation of oil biodegradation was also demonstrated in 25 nutrient-poor lakes of northern Wisconsin, although in almost every case oil-degrading bacteria were detected. Knowledge of temperature and nutrient limitations thus will help in predicting the fate of hydrocarbon pollutants in freshwater.     

Characterization of microaerophilic bacteria isolated from the coastal waters of Tokyo Bay, Japan

Abstract A total of 84 strains of microaerophiles were isolated from the Edo River mouth, Tokyo Bay, Japan, and their morphological, physiological and biochemical properties and mole percent guanine-plus-cytosine contents (mol%) of DNAs extracted from them were examined. Eighty-eight to 100% of the strains, collected in summer, grew at 15–37°C but only 5% of the strains grew at 5°C. Seventy-seven to 100% of the strains, collected in winter, were able to grow at 10–25°C but 79% of the strains were unable to grow at 37°C. The shift in growth-range of bacteria strongly suggests that the microflora of estuarine microaerophiles changes seasonally. All the strains could grow at 50% seawater while 70 and 42% of the strains could not grow at 0 and 100% seawater, respectively.
On the other hand, 72 strains, out of the 84 strains, clustered into 9 phena using numerical taxonomy methods. The strains belonging to phena 1 through 9 were all Gram-negative, non-spore-forming rod-shaped organisms which were negative for susceptibility to vibrostatic agent O/129, H₂S production and acid from dulcitol, and the G + C contents of DNAs ranged from 29.8 to 56.6 mol%. However, almost all strains could not be identified and are suggested to be new species.     

Distribution and significance of heterotrophic marine bacteria with antibacterial activity

Bacteria with antibacterial activity were isolated from seawater, sediments, phytoplankton, and zooplankton of Suruga, Sagami, and Tokyo Bays and from soft corals and sponges collected from the Taiwan coast. Of the 726 strains isolated, 37 showed antibacterial activity against either Vibrio parahaemolyticus (ATCC 17802) or Staphylococcus aureus (P209). Sediment harbored the lowest number of these forms of bacteria, and those from Tokyo Bay did not show any activity. Attached isolates showed greater activity compared with free-living forms. Relatively high numbers of strains with antibacterial activity were associated with phytoplankton. Among the zooplankton isolates, cladocerans harbored the maximum number of antibacterial strains. Isolates were more inhibitory to gram-positive test cultures. Autoinhibition was observed only among 8% of the isolates. Marine nonproducers were more susceptible. Pseudomonas/Alteromonas species made up 81.0% of isolates, of which 30% were pigmented strains. The absence or reduction in number of bacteria with antibacterial activity in Tokyo Bay is attributed to its eutrophic nature, which may tend to moderate the production of antibacterial compounds.     

Distribution and significance of heterotrophic marine bacteria with antibacterial activity.

Bacteria with antibacterial activity were isolated from seawater, sediments, phytoplankton, and zooplankton of Suruga, Sagami, and Tokyo Bays and from soft corals and sponges collected from the Taiwan coast. Of the 726 strains isolated, 37 showed antibacterial activity against either Vibrio parahaemolyticus (ATCC 17802) or Staphylococcus aureus (P209). Sediment harbored the lowest number of these forms of bacteria, and those from Tokyo Bay did not show any activity. Attached isolates showed greater activity compared with free-living forms. Relatively high numbers of strains with antibacterial activity were associated with phytoplankton. Among the zooplankton isolates, cladocerans harbored the maximum number of antibacterial strains. Isolates were more inhibitory to gram-positive test cultures. Autoinhibition was observed only among 8% of the isolates. Marine nonproducers were more susceptible. Pseudomonas/Alteromonas species made up 81.0% of isolates, of which 30% were pigmented strains. The absence or reduction in number of bacteria with antibacterial activity in Tokyo Bay is attributed to its eutrophic nature, which may tend to moderate the production of antibacterial compounds.     

Comparative study of the aerobic, heterotrophic bacterial flora of Chesapeake Bay and Tokyo Bay.

A comparative study of the bacterial flora of the water of Chesapeake Bay and Tokyo Bay was undertaken to assess similarities and differences between the autochthonous flora of the two geographical sites and to test the hypothesis that, given similarities in environmental parameters, similar bacterial populations will be found, despite extreme geographic distance between locations. A total of 195 aerobic, heterotrophic bacterial strains isolated from Chesapeake Bay and Tokyo Bay water were examined for 115 biochemical, cultural, morphological, nutritional, and physiological characters. The data were analyzed by the methods of numerical taxonomy. From sorted similarity matrices, 77% of the isolates could be grouped into 30 phena and presumptively identified as Acinetobacter-Moraxella, Caulobacter, coryneforms, Pseudomonas, and Vibrio spp. Vibrio and Acinetobacter species were found to be common in the estuarine waters of Chesapeake Bay, whereas Acinetobacter-Moraxella and Caulobacter predominated in Tokyo Bay waters, at the sites sampled in the study.     

Comparative study of the aerobic, heterotrophic bacterial flora of Chesapeake Bay and Tokyo Bay.

A comparative study of the bacterial flora of the water of Chesapeake Bay and Tokyo Bay was undertaken to assess similarities and differences between the autochthonous flora of the two geographical sites and to test the hypothesis that, given similarities in environmental parameters, similar bacterial populations will be found, despite extreme geographic distance between locations. A total of 195 aerobic, heterotrophic bacterial strains isolated from Chesapeake Bay and Tokyo Bay water were examined for 115 biochemical, cultural, morphological, nutritional, and physiological characters. The data were analyzed by the methods of numerical taxonomy. From sorted similarity matrices, 77% of the isolates could be grouped into 30 phena and presumptively identified as Acinetobacter-Moraxella, Caulobacter, coryneforms, Pseudomonas, and Vibrio spp. Vibrio and Acinetobacter species were found to be common in the estuarine waters of Chesapeake Bay, whereas Acinetobacter-Moraxella and Caulobacter predominated in Tokyo Bay waters, at the sites sampled in the study.     

The enhancement by surfactants of hexadecane degradation by Pseudomonas aeruginosa varies with substrate availability.

The rhamnolipid biosurfactant produced by Pseudomonas aeruginosa influences various processes related to hydrocarbon degradation. However, degradation can only be enhanced by the surfactant when it stimulates a process that is rate limiting under the applied conditions. Therefore we determined how rhamnolipid influences hexadecane degradation by P. aeruginosa UG2 under conditions differing in hexadecane availability. The rate of hexadecane degradation in shake flask cultures was lower for hexadecane entrapped in a matrix with 6 nm pores (silica 60) or in quartz sand than for hexadecane immobilized in matrices with pore sizes larger than 300 nm or for hexadecane present as a separate liquid phase. This indicates that the availability of hexadecane decreased with decreasing pore size under these conditions. The rate-limiting step for hexadecane entrapped in silica 60 was the mass transfer of substrate from the matrix to the bulk liquid phase, whereas for hexadecane present as a second liquid phase it was the uptake of the substrate by the cells. Hexadecane degradation in batch incubations was accelerated by the addition of rhamnolipid or other surfactants in all experiments except in those where hexadecane was entrapped in silica 60, indicating that the surfactants stimulated uptake of hexadecane by the cells. Since rhamnolipid stimulated the degradation rate in batch experiments to a greater extent than any of the other 14 surfactants tested, hexadecane uptake was apparently more enhanced by rhamnolipid than by the other surfactants. Although rhamnolipid did not stimulate the release of hexadecane from silica 60 under conditions of intense agitation, it significantly enhanced this rate during column experiments in the absence of strain UG2. The results demonstrate that rhamnolipid enhances degradation by stimulating release of entrapped substrate in column studies under conditions of low agitation and by stimulating uptake of substrate by the cells, especially when degradation is not limited by release of substrate from the matrices.     

Productivity and grazing impact of Oikopleura dioica (Tunicata, Appendicularia) in Tokyo Bay

To estimate the productivity and grazing impact of a commonappendicularian species Oikopleura dioica in Tokyo Bay, monthlyobservations on its abundance and vertical distribution wereconducted during 2000. The abundance peaked in February andOctober, but was low during summer. Seasonal fluctuations inproductivity were similar to those of the abundance, with maximumvalues of 92, 134 and 508 mg C m^–2 day^–1 for somatic,new house and discarded house productivity in October and 206mg C m^–2 day^–1 for fecal pellet productivity inFebruary, respectively. The averaged biomass of O. dioica wasonly 2.5% of that of copepods; however, the secondary productivityof the former corresponded to 12.4% of the latter. Daily grazingimpact on particulate organic carbon ranged from 0.05% to 5%,which is close to the impact by copepods. These results implythat in Tokyo Bay, where small copepods and jellyfish are abundant,O. dioica is an important component of the ecosystem becauseit bridges between small primary producers and higher consumers.     

Effect of environmental factors on the relationship between concentrations of coprostanol and fecal indicator bacteria in tropical (Mekong Delta) and temperate (Tokyo) freshwaters

A reliable assessment of microbial indicators of fecal pollution (total coliform, Escherichia coli, and fecal streptococcus) is critical in tropical environments. Therefore, we investigated the relationship between concentrations of indicator bacteria and a chemical indicator, coprostanol (5beta-cholestan-3beta-ol), in tropical and temperate regions. Water samples were collected from the Mekong Delta, Vietnam, during wet and dry seasons, and from Tokyo, Japan, during summer, the aftermath of a typhoon, and winter. During the wet season in the Mekong Delta, higher bacterial densities were observed in rivers, probably due to the higher bacterial inputs from soil particles with runoff. In Tokyo, higher bacterial densities were usually observed during summer, followed by those in the typhoon aftermath and winter. A strong logarithmic correlation between the concentrations of E. coli and coprostanol was demonstrated in all surveys. Distinctive seasonal fluctuations were observed, as concentrations of coprostanol corresponding to 1,000 CFU of E. coli/100 ml were at their lowest during the wet season in the Mekong Delta and the typhoon aftermath in Tokyo (30 ng/liter), followed by the dry season in the Mekong Delta and the summer in Tokyo (100 ng/liter), and they were much higher during the winter in Tokyo (400 ng/liter). These results suggested that E. coli is a specific indicator of fecal contamination in both tropical and temperate regions but that the densities are affected by elevated water temperature and input from runoff of soil particles. The concurrent determination of E. coli and coprostanol concentrations could provide a possible approach to assessing the reliability of fecal pollution monitoring data.     

全球四大湾区生态环境综合评价研究

全球四大湾区既是经济发达区,也是生态环境脆弱区,对全球可持续发展至关重要。粤港澳大湾区的生态环境问题正在限制其可持续发展,有碍于全面建成国际一流湾区目标的实现。然而,过去的研究对生态环境方面关注较少,缺少湾区生态环境综合评价体系的构建。通过综合国内外统计和遥感多源数据,建立四大湾区生态环境评价数据库;围绕生态质量,环境质量和资源利用效率建立指标体系,构建湾区生态环境综合评价指数。结果表明：2015年,生态环境综合评价,东京湾区 > 旧金山湾区 > 纽约湾区 > 粤港澳大湾区;生态质量,纽约湾区 > 旧金山湾区 > 粤港澳大湾区 > 东京湾区;环境质量,旧金山湾区 > 东京湾区 > 纽约湾区 > 粤港澳大湾区;资源利用效率,东京湾区 > 旧金山湾区 > 纽约湾区 > 粤港澳大湾区。2005-2015年,粤港澳大湾区环境质量增速缓慢,在生态质量和资源利用效率增幅排名首位。旧金山湾区和纽约湾区优势在生态和环境质量,短板在资源利用效率;东京湾区在资源利用效率方面优势明显,短板在生态质量;综合来看,粤港澳大湾区与其他湾区都存在差距。研究建议粤港澳大湾区从生态空间、生态环境治理和资源利用方面,吸取发达国家湾区的经验。     

UAF radiorespirometric protocol for assessing hydrocarbon mineralization potential in environmental samples

Following the EXXOn Valdez oil spill, a radiorespirometric protocol was developed at the University of Alaska Fairbanks (UAF) to assess the potential for microorganisms in coastal waters and sediments to degrade hydrocarbons. The use of bioremediation to assist in oil spill cleanup operations required microbial bioassays to establish that addition of nitrogen and phosphorus would enhance biodegradation. A technique assessing 1-¹⁴C-n-hexadecane mineralization in seawater or nutrient rich sediment suspensions was used for both of these measurements. Hydrocarbon-degradation potentials were determined by measuring mineralization associated with sediment microorganisms in sediment suspended in sterilized seawater and/or marine Bushnell-Haas broth. Production of ¹⁴CO₂ and CO₂ was easily detectable during the first 48 hours with added hexadecane levels ranging from 10 to 500 mg/l of suspension and dependent on the biomass of hydrocarbon degraders, the hydrocarbon-oxidation potential of the biomass and nutrient availability. In addition to assessment of the hydrocarbon-degrading potential of environmental samples, the radiorespirometric procedure, and concomitant measurement of microbial biomass, has utility as an indicator of hydrocarbon contamination of soils, aqueous sediments and water, and can also be used to evaluate the effectiveness of bioremediation treatments.     

Occurrence, seasonality and genetic diversity of Vibrio vulnificus in coastal seaweeds and water along the Kii Channel, Japan

Vibrio vulnificus is a ubiquitous toxigenic bacterium found in a coastal environment but little is known about its occurrence and seasonality among seaweeds, which are widely consumed as seafood in Japan. Therefore, we have observed the bacterium's abundance in seawater and seaweed samples from three areas of the Kii Channel, Japan, during June 2003 to May 2004. A total of 192 samples were collected: 24 from each source in summer, autumn, winter and spring. The samples were selectively cultivated following the most probable number (MPN) technique. Vibrio vulnificus population ranged from 0 to 10(3) MPN 100 mL(-1) seawater or 10 g seaweeds; higher counts were observed during summer. The optimum temperature, salinity and pH for the bacterium were 20-24 degrees C, 24-28 p.p.t. and 7.95-8.15, respectively. However, seaweeds always contained higher V. vulnificus than seawater. Among 280 V. vulnificus strains, detected by species-specific colony hybridization and PCR, 78, 74, 11 and 16 were from seaweeds and 46, 42, 2 and 11 were from seawater during summer, autumn, winter and spring, respectively. Ribotyping of 160 selected strains revealed a higher genotypic diversity (18 patterns) among strains from seaweeds than from seawater (10 patterns). Seaweeds can thus act as a potential habitat for V. vulnificus and are more unsafe for consumption during summer.