Number of bacteria and features of their activity in hypersaline reservoirs of the Crimea

The incidence of bacteria, their biomass production, and heterotrophic assimilation of CO2 by bacterioplankton were studied in the Crimean hypersaline water reservoirs from May to October of 1974. The total incidence of bacteria in the natural brine of these reservoirs varied from 20 to 70 x 10(6) cells per 1 ml. Such a high bacterial number may be caused by the combined action of water evaporation which increased the concentration of bacterial cells and active growth of microflora. Low values of bacterial production and heterotrophic CO2 assimilation should be attributed to weak activity of microflora in the reservoirs.     

Taxonomic composition of bacteria associated with cultivated mollusks Crassostrea lugubris and Perna viridis and with the water of the Gulf of Nha Trang Lagoon, Vietnam

One hundred and four strains of heterotrophic bacteria have been isolated and characterized from two species of bivalve mollusks cultivated in the Gulf of Nha Trang (Vietnam) and from the water of a mariculture farm. The isolates have been identified on the basis of morphological, physiological, biochemical, and chemotaxonomic properties, as well as by the content of G+C bases in DNA. In the microflora of mollusks, Vibrio alginolyticus was predominant; the pathogenic species V. harveyi and V. splendidus were found as well. Staphylococci and bacilli occupied the second place in abundance after vibrios. In addition, coryneforms and enterobacteria, as well as Pseudomonas spp. and Pseudoalteromonas spp., were revealed. The composition of the water microflora was more diverse as compared with the microflora of mollusks. In the water, Bacillus spp., Vibrio spp., and Pseudomonas spp. were predominant. Brevibacterium spp. and other coryneform bacteria, as well as enterobacteria, occurred in significant amounts. In addition, Pseudoalteromonas spp., Marinococcus sp., Halobacillus sp., Shewanella sp., Sulfitobacter sp., and bacteria of the CFB cluster were noticed. The presence of pathogenic and conditionally pathogenic bacterial species in the water and mollusks is probably the reason for the high death rate of cultivated animals at the mariculture farm.     

Characterization of communities of heterotrophic bacteria associated with healthy and diseased corals in Nha Trang Bay (Vietnam)

A comparative investigation of the heterotrophic microflora of 11 species of healthy corals and of white-band-diseased and yellow-band-diseased corals inhabiting the reefs of Nha Trang Bay (Vietnam), which has been exposed to anthropogenic impact, was performed. Fifty-nine strains of heterotrophic bacteria isolated on Y/K and Endo media were investigated and characterized. All the isolates were identified at the genus level by consideration of the results of analysis of their phenotypic properties, determination of the molar percent of G+C bases in their DNA, and the composition of fatty acids of their lipids. In the composition of the microflora of tissues of healthy corals, gamma-proteobacteria prevailed, with halomonads being dominant among them. In addition, the gram-negative bacteria included Pseudomonas and Vibrio spp., members of the Cytophaga-Flavobacterium-Bacteroides (CFB) phylogenetic cluster, and Moraxella sp. The gram-positive bacteria revealed included Bacillus, Staphylococcus, Halococcus, and Micrococcus spp., and coryneform bacteria. In the composition of the microflora of the tissues of affected corals, bacteria of the family Enterobacteriaceae and of the genera Planococcus and Arthrobacter, which were not revealed in healthy hydrobionts, were found. The anthropogenic impact is not the sole factor determining the infection of corals.     

Geographical and seasonal distribution of multiple antibiotic resistance of heterotrophic bacteria of Lake Shira

From 1996 to 1999 heterotrophic bacteria of the brackish-water Lake Shira (Republic of Khakasia, Russia) were studied to understand the seasonal dynamics of their antibiotic resistance. During the winter, these bacteria were represented primarily by forms that could not be cultured and were psychrotolerant. In the summer period, heterotrophic, mesophilic bacteria increased in number. The percentages of isolates with multiple, antibiotic resistance isolated from the lake region near the resort area of the lake were 2–3 times higher than those from the central part of the lake. A decline in the bacterial numbers with multiple antibiotic resistance was observed during the cold period (February–March). Various mechanisms of multiple, antibiotic resistance of heterotrophic bacteria isolated from Shira lake are discussed.     

Characterization of Communities of Heterotrophic Bacteria Associated with Healthy and Diseased Corals in Nha Trang Bay (Vietnam)

A comparative investigation of the heterotrophic microflora of 11 species of healthy corals and of white-band-diseased and yellow-band-diseased corals inhabiting the reefs of Nha Trang Bay (Vietnam), which has been exposed to anthropogenic impact, was performed. Fifty-nine strains of heterotrophic bacteria isolated on Y/K and Endo media were investigated and characterized. All the isolates were identified at the genus level by consideration of the results of analysis of their phenotypic properties, determination of the molar percent of G+C bases in their DNA, and the composition of the fatty acids of lipids. In the composition of the microflora of tissues of healthy corals, γ-proteobacteria prevailed, with halomonads being dominant among them. In addition, the gram-negative bacteria included Pseudomonas and Vibrio spp., members of the Cyto-phaga-Fla-vobacterium-Bacteroides (CFB) phylogenetic cluster, and Moraxella sp. The gram-positive bacteria revealed included Bacillus, Staphylococcus, Halococcus, and Micrococcus spp., and coryneform bacteria. In the composition of the microflora of the tissues of affected corals, bacteria of the family Enterobacteriaceae and of the genera Planococcus and Arthrobacter, which were not revealed in healthy hydrobionts, were found. The anthropogenic impact is not the sole factor determining the infection of corals.     

Effect of low-roughage diets on the microflora and lipid metabolism in the rumen

Changing the diet of five lactating cows and one nonlactating cow from high to low roughage induced milk fat depression in the lactating cows and altered the composition of the rumen microflora. While the numbers of lactic and propionic acid-producing bacteria increased, the numbers of Butyrivibrio spp. decreased. The numbers of lipolytic bacteria and the in vitro lipolytic activity of the rumen fluid were also decreased, as was the extent of hydrogenation of linoleic and linolenic acids combined in soybean oil incubated in vitro with rumen fluid. It is suggested that among the bacterial population in the rumen the vibrios, which were adversely affected by the low-roughage diets, may contribute significantly to both lipolysis and hydrogenation in the rumen.     

Isolation and enumeration of marine actinomycetes from seawater and sediments in Alexandria

Actinomycetes were isolated from near-shore marine sediments and water at four different sites in Alexandria. Statistical analysis revealed that variation in temperature, pH, and dissolved phosphate were of insignificant values, but that variation in total nitrogen and organic matter were significant. The treatment of sediments and water samples by heat resulted in a selective reduction of the nonactinomycetal heterotrophic microflora. Four selective culture media were used for counting actinomycetes in marine water and sediments. The starch nitrate medium favored the growth of these microorganisms. The diversity and counts of actinomycetes varied with the seasonal variation, and the highest counts were detected in dry warm seasons. The numbers of this bacterial group in sediments exceeded by far their numbers in seawater. A positive correlation was found between population size and location. Actinomycetes were found in the highest numbers in the upper layers (0-20 cm depth). In a few cases, the counts of actinomycetes showed bimodal maxima 0-20 and 60-100 cm deep. Sediments were the best source of marine actinomycetes, and their distribution varied depending on the depth from which samples were collected. The ratio of actinomycetes to the total microflora ranged from 0.48 to 2.29, depending on location.     

基于高通量测序的缢蛏及其养殖池塘菌群结构的季节变化

为探究缢蛏养殖过程中养殖环境和缢蛏体内的细菌群落结构季节变化特征及其与环境因子的相关性,采用高通量测序技术对缢蛏养殖池塘的水体、沉积物及缢蛏内脏团细菌菌群进行了研究.结果表明: 水体中冬季的细菌菌群结构与其他3季差异显著,沉积物和缢蛏内脏团4季的细菌菌群结构均无显著差异.水体各季节细菌群落的Shannon多样性指数无明显差异,沉积物及缢蛏内脏团细菌群落Shannon多样性指数夏季最低,冬季最高.门水平上,蓝细菌门、变形菌门和拟杆菌门为水体的优势菌群;变形菌门、绿弯菌门和拟杆菌门为沉积物的优势菌群;柔膜菌门和变形菌门为缢蛏内脏团的优势菌群.属水平上,水体冬季的优势菌属为NS3a_marine_group,而其余3季为聚球藻属;沉积物优势菌属为厌氧绳菌科下的一类菌属norank_f_Anaerolineacea和硝化螺旋菌属;内脏团优势菌属为支原体属和弓形菌属.环境因子与菌群的相关性分析表明: 水体的优势菌属聚球藻属与水温、化学需氧量(COD)、PO₄^--P、NH₄⁺-N、pH和透明度呈正相关;沉积物的优势菌属厌氧绳菌科菌属norank_f_Anaerolineacea与COD、水温、总磷呈正相关;缢蛏内脏团的优势菌属支原体属与水温、pH、NH₄⁺-N、PO₄^--P和透明度呈正相关.这表明不同季节的缢蛏养殖池塘及缢蛏体内细菌群落结构及多样性差异较大,水体细菌菌群受养殖环境的影响明显,尤其是水温及氮磷含量.     

Heterotrophic bacteria accompanying Stichococcus bacillaris in laboratory cultures

The number and composition of bacterial microflora accompanying Stichococcus bacillaris in various media with urea was determined. The number of heterotrophic bacteria during 5 days of incubation increased 10-fold. Only some of the isolated bacterial strains were able to grow in medium for algae supplemented with dead S. bacillaris cells. It is suggested that bacteria utilize organic matter released to the medium by S. bacillaris. The most numerous among the isolated bacteria were Enterobacteriaceae. The order of domination by bacterial populations distinguishes nonaxenic cultures of S. bacillaris from cultures of other algae.     

On-board flow cytometric observation of picoplankton community structure in the East China Sea during the fall of different years

On-board flow cytometric determinations of picoplankton abundance (i.e. Synechococcus spp., Prochlorococcus spp., picoeukaryotes and also heterotrophic bacteria) were obtained in the East China Sea in fall of 2000 and 2003. The average abundances of Synechococcus, Prochlorococcus, picoeukaryotes and heterotrophic bacteria were 10(5), 10(5), 10(4) and 10(6) cells ml(-1), respectively. Synechococcus, picoeukaryotes and heterotrophic bacteria were abundant at all the stations and presented higher concentration in the inner shelf where influences from the Changjiang effluent plumes and the coastal upwelling were evident, while Prochlorococcus was absent from the near-shore stations and became the dominant picophytoplankton population in offshore waters, where its abundance was comparable to that for heterotrophic bacteria. All picoplankton groups showed a reduction in cell number with depth, and a positive correlation with water temperature were observed, which reflected the importance of light and temperature on picoplankton growth. A negative relationship with salinity was found for heterotrophic bacteria along two sections across the East China Sea Shelf, and distribution of picoplankton was dominated by different water masses. The fixation could lead to loss in Prochlorococcus cell numbers within one month, and all the picoplankton numbers decreased dramatically after three months.     

Taxonomic composition of bacteria associated with cultivated mollusks <Emphasis Type="Italic">Crassostrea lugubris</Emphasis> and <Emphasis Type="Italic">Perna viridis</Emphasis> and with the water of the Gulf of Nha Trang lagoon,Vietnam

One hundred and four strains of heterotrophic bacteria have been isolated and characterized from two species of bivalve mollusks cultivated in the Gulf of Nha Trang (Vietnam) and from the water of a mariculture farm. The isolates have been identified on the basis of morphological, physiological, biochemical, and chemotaxonomic properties, as well as by the content of G+C bases in DNA. In the microflora of mollusks, Vibrio alginolyticus was predominant; the pathogenic species V. harveyi and V. splendidus were found as well. Staphylococci and bacilli occupied the second place in abundance after vibrios. In addition, coryneforms and enterobacteria, as well as Pseudomonas spp. and Pseudoalteromonas spp., were revealed. The composition of the water microflora was more diverse as compared with the microflora of mollusks. In the water, Bacillus spp., Vibrio spp., and Pseudomonas spp. were predominant. Brevibacterium spp. and other coryneform bacteria, as well as enterobacteria, occurred in significant amounts. In addition, Pseudoalteromonas spp., Marinococcus sp., Halobacillus sp., Shewanella sp., Sulfitobacter sp., and bacteria of the CFB cluster were noticed. The presence of pathogenic and conditionally pathogenic bacterial species in the water and mollusks is probably the reason for the high death rate of cultivated animals at the mariculture farm.     

Diversity and dynamics of a north atlantic coastal Vibrio community

Vibrios are ubiquitous marine bacteria that have long served as models for heterotrophic processes and have received renewed attention because of the discovery of increasing numbers of facultatively pathogenic strains. Because the occurrence of specific vibrios has frequently been linked to the temperature, salinity, and nutrient status of water, we hypothesized that seasonal changes in coastal water bodies lead to distinct vibrio communities and sought to characterize their level of differentiation. A novel technique was used to quantify shifts in 16S rRNA gene abundance in samples from Barnegat Bay, N.J., collected over a 15-month period. Quantitative PCR (QPCR) with primers specific for the genus Vibrio was combined with separation and quantification of amplicons by constant denaturant capillary electrophoresis (CDCE). Vibrio populations identified by QPCR-CDCE varied between summer and winter samples, suggesting distinct warm-water and year-round populations. Identification of the CDCE populations by cloning and sequencing of 16S rRNA genes from two summer and two winter samples confirmed this distinction. It further showed that CDCE populations corresponded in most cases to approximately 98% rRNA similarity groups and suggested that the abundance of these follows temperature trends. Phylogenetic comparison yielded closely related cultured and often pathogenic representatives for most sequences, and the temperature ranges of these isolates confirmed the trends seen in the environmental samples. Overall, this suggests that temperature is a good predictor of the occurrence of closely related vibrios but that considerable microdiversity of unknown significance coexists within this trend.     

Profiling bacterial survival through a water treatment process and subsequent distribution system

AIMS: To profile fractions of active bacteria and of bacteria culturable with routine heterotrophic plate count (HPC) methods through a typical water treatment process and subsequent distribution system. In doing so, investigate how water treatment affects both bacterial abundance and diversity, and reveal the identities of active bacteria not detected by traditional HPC culture. METHODS AND RESULTS: Profiling active fractions was performed by flow cytometric cell sorting of either membrane-intact (BacLight kit) or enzymatically active (carboxyfluorescein diacetate, CFDA) bacteria, followed by eubacterial 16S rDNA-directed PCR and denaturing gradient gel electrophoresis (DGGE). Water treatment significantly reduced active bacterial numbers detected by the BacLight kit and CFDA assay by 2.89 and 2.81 log respectively. Bacterial diversity was also reduced from > 20 DGGE bands in the active fractions of reservoir water to only two bands in the active fractions of finished water. These two bands represented Stenotrophomonas maltophila, initially culturable by HPC, and a Burkholderia-related species. Both species maintained measurable traits of physiological activity in distribution system bulk water but were undetected by HPC. CONCLUSIONS: Flow cytometric cell sorting with PCR-DGGE, to assess water treatment efficacy, identified active bacteria from a variety of major phylogenetic groups undetected by routine HPC. Following treatment S. maltophila and a Burkholderia-related species retained activity and entered distribution undetected by HPC. SIGNIFICANCE AND IMPACT OF THE STUDY: Methods used here demonstrate how water treatment operators can better monitor water treatment plant efficacy and assess distribution system instability by the detection and identification of active bacteria recalcitrant to routine HPC culture.     

Activity of Microorganisms in Acid Mine Water I. Influence of Acid Water on Aerobic Heterotrophs of a Normal Stream

Comparison of microbial content of acid-contaminated and nonacid-contaminated streams from the same geographical area indicated that nonacid streams contained relatively low numbers of acid-tolerant heterotrophic microorganisms. The acid-tolerant aerobes survived when acid entered the stream and actually increased in number to about 2 × 10³ per ml until the pH approached 3.0. The organisms then represented the heterotrophic aerobic microflora of the streams comprised of a mixture of mine drainage and nonacid water. A stream which was entirely acid drainage did not have a similar microflora. Most gram-positive aerobic and anaerobic bacteria died out very rapidly in acidic water, and they comprised a very small percentage of the microbial population of the streams examined. Iron- and sulfur-oxidizing autotrophic bacteria were present wherever mine water entered a stream system. The sulfur-oxidizing bacteria predominated over iron oxidizers. Ecological data from the field were verified by laboratory experiments designed to simulate stream conditions.     

Abundance,biomass, and production of heterotrophic bacteria in a large plain reservoir during the ice-covered period

The abundance, biomass, production, and size and morphological structure of heterotrophic bacteria have been determined and their contribution to the total plankton biomass has been assessed in the Rybinsk Reservoir (the Upper Volga) during the ice-covered period. Structural and functional characteristics of bacteria, except for their cell sizes in winter, are lower than those in the growing season. However, heterotrophic bacteria have made the main contribution (89.3%) to the total biomass of plankton community in winter. During the growing season, phytoplankton is a major component of plankton (48.8%), while the bacteria average 36.5%. The total plankton biomass under the ice averages 148 mg C/m³, which is 2.6 times lower than in the period of open water. During the ice-covered period, the number of protists is small and they consume an insignificant part of the bacterial production; viral lysis is the main reason for bacterial mortality.     

Effect of Bacillus thuringiensis and a pyrethroid insecticide on the leaf microflora of Brassica oleracea

In a field experiment comparing the effect of treatment with Bacillus thuringiensis (Bt) and the pyrethroid insecticide, cypermethrin, on leaf microflora across the growing season, no significant differences were found, apart from more bacteria on cypermethrin-treated leaves on 24 September ( P < 0·05). No effect was seen on fungal diversity or numbers. In contrast, the pyrethroid insecticide inhibited growth of 44% of bacterial isolates tested in vitro; Bt was not inhibitory. Bacillus thuringiensis numbers on treated leaves increased throughout the season, following repeated applications.     

Influence of pipe materials and VBNC cells on culturable bacteria in a chlorinated drinking water model system

To elucidate the influence of pipe materials on the VBNC (viable but nonculturable) state and bacterial numbers in drinking water, biofilm and effluent from stainless steel, galvanized iron, and polyvinyl chloride pipe wafers were analyzed. Although no HPC (heterotrophic plate count) was detected in the chlorinated influent of the model system, a DVC (direct viable count) still existed in the range between 3- and 4-log cells/ml. Significantly high numbers of HPC and DVC were found both in biofilm and in the effluent of the model system. The pipe material, exposure time, and the season were all relevant to the concentrations of VBNC and HPC bacteria detected. These findings indicate the importance of determining the number of VBNC cells and the type of pipe materials to estimate the HPC concentration in water distribution systems and thus the need of determining a DVC in evaluating disinfection efficiency.     

First assessment of microbial diversity in faecal microflora of Eurasian otter (<Emphasis Type="Italic">Lutra lutra</Emphasis> Linnaeus, 1758) in Portugal

The Eurasian otter (Lutra lutra Linnaeus, 1758), a predator from the Order Carnivora, Family Mustelidae, evolved the ability to swim and forage in water, being an important element of biodiversity. Otters are widely spread through Portugal, and scats have been extensively used in ecology studies; however, valid information on their microbiota is scarce. This work represents a first approach to characterise the otter faecal microflora in samples collected in river stretches of the Sado river basin (Portugal) during winter 2006. Eight sampling stretches of 8 km were selected, and from each, six to eight sampling sites were visited. A total of 31 scats were analysed. The microflora studied included aerobic bacteria, spore-forming anaerobic bacteria and viruses (coronavirus, parvovirus, adenovirus, parainfluenza virus). Bacterial isolates were identified based on morphology and metabolic pathways, and virus detection was performed by polymerase chain reaction. The results revealed the high degree of bacterial diversity in the faecal microflora of L. lutra. A total of 88 Gram-negative (23 genera) and 44 Gram-positive isolates (ten genera) were identified. The identification of four isolates was inconclusive, and their identification was performed by 16S ribosomal ribonucleic acid sequencing, which confirms the need for biochemical testing optimisation regarding animal isolates. None of the scats was positive for virus detection. Identification of otter faecal microflora and of potential pathogens is an important first step towards understanding and monitoring their importance in otter population health.     

Occurrence of potentially pathogenic vibrios and related environmental factors in Songkhla Lake, Thailand

Vibrios are halophilic bacteria that are ubiquitous in marine environments. Their occurrence in tropical lakes has rarely been investigated. In this study, the predominance and diversity of Vibrio spp. was investigated over a 12-month period in a coastal lagoon, Songkhla Lake, in southern Thailand. Water samples were collected at 2 stations in the estuary near Yor Island in Songkhla Lake. The predominant vibrios were detected by a culture-based method, using thiosulfate-citrate-bile salt-sucrose agar and CHROMagar Vibrio. The diversity of Vibrio spp. was evaluated using denaturant density gradient electrophoresis (DGGE). The highest numbers of total vibrios and Vibrio parahaemolyticus in both areas were observed during the summer. There was no significant correlation between the numbers of vibrios, including V. parahaemolyticus, and either the water temperature or plankton density. Variations in Vibrio species were observed with changes in salinity. Vibrio parahaemolyticus and V. cholerae non-O1/non-O139 were detected during the rainy season when the salinity dropped to nearly 0 parts per thousand. In both areas, V. alginolyticus was the most prominent species detected by the culture method, whereas Vibrio parahaemolyticus was detected by DGGE, every month. Other Vibrio spp. of potential public health concern were also detected by the culture method; they included V. vulnificus , V. fluvialis , and V. mimicus .     

A method of the selective isolation and enumeration of marine Vibrionaceae

A noninhibitory medium and GasPack anaerobic culture system were employed for the selective enumeration and isolation of Vibrionaceae in seawater and marine sediments.Vibrio counts obtained by the new method for seawater and sediment samples were compared with vibrio numbers in the heterotrophic bacterial population appearing on a medium routinely employed in the laboratory for such counts. The ratio of the former to the latter counts ranged from 0.5 to 1.3, the average being 0.96. The seawater and sediment bacteria that grew and produced visible colonies on the medium under anaerobic conditions for 3 days at 20°C were almost exclusively vibrios.From the results reported here it is concluded that most of the vibrios present in seawater and sediment samples can be recovered by the new method developed in this study.