Luminous Enteric Bacteria of Marine Fishes: a Study of Their Distribution, Densities, and Dispersion

Three taxa of luminous bacteria (Photobacterium fischeri, P. phosphoreum, and Beneckea spp.) were found in the enteric microbial populations of 22 species of surface- and midwater-dwelling fishes. These bacteria often occurred in concentrations ranging between 10⁵ and 10⁷ colony-forming units per ml of enteric contents. By using a genetically marked strain, it was determined that luminous cells entering the fish during ingestion of seawater or contaminated particles traversed the alimentary tract and survived the digestive processes. After excretion, luminous bacteria proliferated extensively on the fecal material and became distributed into the surrounding seawater. Thus, this enteric habitat may serve as an enrichment of viable cells entering the planktonic luminous population.     

Distribution and Identification of Luminous Bacteria from the Sargasso Sea

Vibrio fischeri and Lucibacterium harveyi constituted 75 of the 83 luminous bacteria isolated from Sargasso Sea surface waters. Photobacterium leiognathi and Photobacterium phosphoreum constituted the remainder of the isolates. Luminescent bacteria were recovered at concentrations of 1 to 63 cells per 100 ml from water samples collected at depths of 160 to 320 m. Two water samples collected at the thermocline yielded larger numbers of viable, aerobic heterotrophic and luminous bacteria. Luminescent bacteria were not recovered from surface microlayer samples. The species distribution of the luminous bacteria reflected previously recognized growth patterns; i.e., L. harveyi and V. fischeri were predominant in the upper, warm waters (only one isolate of P. phosphoreum was obtained from surface tropical waters).     

Detection of the Light Organ Symbiont, Vibrio fischeri, in Hawaiian Seawater by Using lux Gene Probes

Symbiotic bacteria that inhabit the light-emitting organ of the Hawaiian squid Euprymna scolopes are distinctive from typical Vibrio fischeri organisms in that they are not visibly luminous when grown in laboratory culture. Therefore, the abundance of these bacteria in seawater samples cannot be estimated simply by identifying them among luminous colonies that arise on nutrient agar plates. Instead, we have used luxR and polymerase chain reaction generated luxA gene probes to identify both luminous and non-visibly luminous V. fischeri colonies by DNA-DNA hybridization. The probes were specific, hybridizing at least 50 to 100 times more strongly to immobilized DNAs from V. fischeri strains than to those of pure cultures of other related species. Thus, even non-visibly luminous V. fischeri colonies could be identified among colonies obtained from natural seawater samples by their probe-positive reaction. Bacteria in seawater samples, obtained either within or distant from squid habitats, were collected on membrane filters and incubated until colonies appeared. The filters were then observed for visibly luminous V. fischeri colonies and hybridized with the lux gene probes to determine the number of total V. fischeri colonies (both luminous and non-visibly luminous). We detected no significant differences in the abundance of luminous V. fischeri CFU in any of the water samples observed (≤1 to 3 CFU/100 ml). However, probe-positive colonies of V. fischeri (up to 900 CFU/100 ml) were found only in seawater collected from within the natural habitats of the squids. A number of criteria were used to confirm that these probe-positive strains were indistinguishable from symbiotic V. fischeri. Therefore, the luxA and luxR gene probes were species specific and gave a reliable estimate of the number of culturable V. fischeri colonies in natural water samples.     

The autumn mesozooplankton community at South Georgia: biomass, population structure and vertical distribution

Mesozooplankton were sampled at shelf and oceanic stations close to South Georgia, South Atlantic during austral autumn 2004 with a Longhurst Hardy Plankton Recorder. Onshelf biomass ranged from 2.18 to 5.75 g DM m^?2 (0–200 m) and was dominated by the small euphausiid Thysanöessa spp. At the oceanic stations (10.57–14.71 g DM m^?2, 0–1,000 m) large calanoids, principally Rhincalanus gigas comprised ～47–52% of biomass. Here Calanus simillimus was still active and reproducing in surface waters (0–11.2 eggs fem day^?1) but R. gigas and Calanoides acutus were largely resident in the warm deep water and undergoing their seasonal descent. A comparison with spring and summer data indicated increased abundance and biomass from spring through to summer followed by a decline towards autumn particularly over the shelf. Autumn values in oceanic waters differed little from summer. Mesozooplankton biomass in the surface 200 m of the oceanic stations as a proportion of that found in the top 1,000 m ranged from 63 to 78% of the total in spring and 62–73% in summer, but was only 23–29% of the total in this study, following redistribution down the water column.     

Aerobic Anoxygenic Phototrophic Bacteria in the Mid-Atlantic Bight and the North Pacific Gyre

The abundance of aerobic anoxygenic phototrophic (AAP) bacteria, cyanobacteria, and heterotrophs was examined in the Mid-Atlantic Bight and the central North Pacific Gyre using infrared fluorescence microscopy coupled with image analysis and flow cytometry. AAP bacteria comprised 5% to 16% of total prokaryotes in the Atlantic Ocean but only 5% or less in the Pacific Ocean. In the Atlantic, AAP bacterial abundance was as much as 2-fold higher than that of Prochlorococcus spp. and 10-fold higher than that of Synechococcus spp. In contrast, Prochlorococcus spp. outnumbered AAP bacteria 5- to 50-fold in the Pacific. In both oceans, subsurface abundance maxima occurred within the photic zone, and AAP bacteria were least abundant below the 1% light depth. The abundance of AAP bacteria rivaled some groups of strictly heterotrophic bacteria and was often higher than the abundance of known AAP bacterial genera (Erythrobacter and Roseobacter spp.). Concentrations of bacteriochlorophyll a (BChl a) were low (~1%) compared to those of chlorophyll a in the North Atlantic. Although the BChl a content of AAP bacteria per cell was typically 20- to 250-fold lower than the divinyl-chlorophyll a content of Prochlorococcus, the pigment content of AAP bacteria approached that of Prochlorococcus in shelf break water. Our results suggest that AAP bacteria can be quite abundant in some oceanic regimes and that their distribution in the water column is consistent with phototrophy.     

Sediment Composition Influences Spatial Variation in the Abundance of Human Pathogen Indicator Bacteria within an Estuarine Environment

Faecal contamination of estuarine and coastal waters can pose a risk to human health, particularly in areas used for shellfish production or recreation. Routine microbiological water quality testing highlights areas of faecal indicator bacteria (FIB) contamination within the water column, but fails to consider the abundance of FIB in sediments, which under certain hydrodynamic conditions can become resuspended. Sediments can enhance the survival of FIB in estuarine environments, but the influence of sediment composition on the ecology and abundance of FIB is poorly understood. To determine the relationship between sediment composition (grain size and organic matter) and the abundance of pathogen indicator bacteria (PIB), sediments were collected from four transverse transects of the Conwy estuary, UK. The abundance of culturable Escherichia coli, total coliforms, enterococci, Campylobacter, Salmonella and Vibrio spp. in sediments was determined in relation to sediment grain size, organic matter content, salinity, depth and temperature. Sediments that contained higher proportions of silt and/or clay and associated organic matter content showed significant positive correlations with the abundance of PIB. Furthermore, the abundance of each bacterial group was positively correlated with the presence of all other groups enumerated. Campylobacter spp. were not isolated from estuarine sediments. Comparisons of the number of culturable E. coli, total coliforms and Vibrio spp. in sediments and the water column revealed that their abundance was 281, 433 and 58-fold greater in sediments (colony forming units (CFU)/100g) when compared with the water column (CFU/100ml), respectively. These data provide important insights into sediment compositions that promote the abundance of PIB in estuarine environments, with important implications for the modelling and prediction of public health risk based on sediment resuspension and transport.     

A Pathogenic Bacterium,Enterococcus faecalis,to the Beet Armyworm,Spodoptera exigua

A bacterial disease was found in the beet armyworm, Spodoptera exigua (Hübner). Blackened body of the infected larvae was a typical symptom of the epizootic disease especially at the intersegmental areas. We isolated the bacteria from the hemolymph of the infected 5th instar larvae and identified the isolate as a gram-positive bacterium, Enterococcus faecalis. When the 4th instar larvae were injected with the bacteria, half lethal dose of the bacteria was estimated as 22,593 colony-forming units (cfu) per larva and half lethal time of the bacteria was estimated as 2 days at 10⁷ cfu injection and 6 days at 10⁸ cfu injection. The bacteria were strongly resistant to each 1,000 ppm of ampicillin, kanamycin, and streptomycin. They were, however, relatively susceptible to mixture (1,000 ppm) of different combinations of the three antibiotics.     

Plants as Sources of Airborne Bacteria, Including Ice Nucleation-Active Bacteria

Vertical wind shear and concentration gradients of viable, airborne bacteria were used to calculate the upward flux of viable cells above bare soil and canopies of several crops. Concentrations at soil or canopy height varied from 46 colony-forming units per m³ over young corn and wet soil to 663 colony-forming units per m³ over dry soil and 6,500 colony-forming units per m³ over a closed wheat canopy. In simultaneous samples, concentrations of viable bacteria in the air 10 m inside an alfalfa field were fourfold higher than those over a field with dry, bare soil immediately upwind. The upward flux of viable bacteria over alfalfa was three- to fourfold greater than over dry soil. Concentrations of ice nucleation-active bacteria were higher over plants than over soil. Thus, plant canopies may constitute a major source of bacteria, including ice nucleation-active bacteria, in the air.     

Effect of Urea Concentration on Growth of Ureaplasma urealyticum (T-Strain Mycoplasma)

The effect of urea on growth of Ureaplasma urealyticum type VIII was studied by cultivating the organisms in a dialysate broth, prepared from soy peptone and autoclaved yeast, supplemented with 5% dialyzed horse serum, 100 mM 2-(N-morpholino)ethane sulfonic acid buffer (pH 5.75), and defined amounts of urea. Without urea, growth did not occur. Total growth was directly related to urea concentration. The least amount of urea that supported growth was 0.032 mM, which resulted in 3 × 10⁴ colony-forming units per ml. The maximum yield of organisms, 8.0 × 10⁷ colony-forming units per ml, was observed at 32 mM urea. Growth was limited not only by urea concentration, but also by the buffer capacity of the medium. The maximum amount of 2-(N-morpholino)ethane sulfonic acid buffer that could be employed was 100 mM; at higher concentrations, growth was inhibited. The yield of U. urealyticum was small even in medium with 32 mM urea and 100 mM 2-(N-morpholino)ethane sulfonic acid buffer: 0.63 mg of protein per liter of culture containing 5 × 10¹⁰ total colony-forming units. The molar growth yield was 20 mg of protein per mol of urea. The growth rate was also a function of urea concentration. Generation times ranged from 8 h at 0.032 mM urea to 1.6 h at 3.2 mM urea, where the substrate level was saturating. The K_s value for growth was 2.0 × 10⁻⁴ M urea. Thus, urea is a growth-limiting factor for U. urealyticum, but remarkably large amounts of this substrate are required.     

Mesozooplankton associations with medium to large marine snow aggregates in the northern Gulf of Mexico

Large and diverse mesozooplankton communities were observedon marine snow particles collected in coastal and oceanic watersof the northern Gulf of Mexico. Mesozooplankton were collectedfrom seven phyla, including ostracods, cladocerans, pelecypodsand ascidian larvae not previ ously recorded as being associatedwith marine snow. Copepod nauplii were the most common, sometimesat concentrations >100 per aggregate. Oncaea spp., Oithonaspp. and Microsetella norvegica were the most common copepodspecies. Total mesozooplankton abundance ranged between 2 and278 organisms per aggregate. Organisms varied markedly in theirdistribution across the aggregate surface and in their behaviourtowards the snow matrix. Comparison of snow communities withzoo-plankton abundance determined from net tows suggests thatsome species are concentrated on snow particles. Snow particlesand their associated microbial communities may be a significantsource of nutrition for these mesozooplankton. Mesozooplanktonmay contribute significantly to the degra dation and decompositionof large snow particles as they sink through the upper watercolumn.     

Detection of Antibody-Coated Bacteria in Expectorated Sputum for Diagnosis of Lower Respiratory Infections

We evaluated antibody-coated bacteria (ACB) in expectorated sputum to discriminate contaminating or colonizing organisms from true pathogens. We examined 60 expectorated sputum samples from 51 patients with lower respiratory infections (chronic obstructive pulmonary disease 25, pneumonia 20, purulent tracheobronchitis 6). All samples were examined with quantitative culture and immunofluorescent demonstration of ACB. From the results of quantitative culture, we divided specimens into pathogen-isolated and pathogen-free samples. Among pathogen-isolated samples, in which we isolated accepted pathogenic organisms at ≥ 10⁷ colony-forming units per ml, 16 of 23 samples were ACB-positive (69.5%). In contrast, among pathogen-free samples, in which we isolated accepted pathogens at < 10⁷ colony forming units per ml or only upper respiratory flora, only 3 of 37 samples were ACB-positive (8.1%). The ACB-positive rate was significantly higher in pathogen-isolated than in pathogen-free samples (P < 0.001). Consequently, detecting ACB in expectorated sputum shows good potential as another criterion for distinguishing contaminating or colonizing organisms from true pathogens.     

Ocean time-series reveals recurring seasonal patterns of virioplankton dynamics in the northwestern Sargasso Sea

There are an estimated 10³⁰ virioplankton in the world oceans, the majority of which are phages (viruses that infect bacteria). Marine phages encompass enormous genetic diversity, affect biogeochemical cycling of elements, and partially control aspects of prokaryotic production and diversity. Despite their importance, there is a paucity of data describing virioplankton distributions over time and depth in oceanic systems. A decade of high-resolution time-series data collected from the upper 300 m in the northwestern Sargasso Sea revealed recurring temporal and vertical patterns of virioplankton abundance in unprecedented detail. An annual virioplankton maximum developed between 60 and 100 m during periods of summer stratification and eroded during winter convective mixing. The timing and vertical positioning of this seasonal pattern was related to variability in water column stability and the dynamics of specific picophytoplankton and heterotrophic bacterioplankton lineages. Between 60 and 100 m, virioplankton abundance was negatively correlated to the dominant heterotrophic bacterioplankton lineage SAR11, as well as the less abundant picophytoplankton, Synechococcus. In contrast, virioplankton abundance was positively correlated to the dominant picophytoplankton lineage Prochlorococcus, and the less abundant alpha-proteobacteria, Rhodobacteraceae. Seasonally, virioplankton abundances were highly synchronous with Prochlorococcus distributions and the virioplankton to Prochlorococcus ratio remained remarkably constant during periods of water column stratification. The data suggest that a significant fraction of viruses in the mid-euphotic zone of the subtropical gyres may be cyanophages and patterns in their abundance are largely determined by Prochlorococcus dynamics in response to water column stability. This high-resolution, decadal survey of virioplankton abundance provides insight into the possible controls of virioplankton dynamics in the open ocean.     

Diversity and characterization of airborne bacteria at two health institutions

The aim of this study was to identify the types and abundance of airborne bacteria of two health institutions (1 and 2) and to determine the genetic association between environmental and clinical isolates of Staphylococcus spp. Environmental sampling in institutions 1 and 2 was conducted for 1 year (dry and rainy seasons) using M Air T sampler. The bacteria and their susceptibility to antibiotics were identified. The colony-forming units per cubic meter (CFU/m³) of air were quantified for all the isolates, and the diversity and abundance of species were calculated. The genetic relationship between the clinical and environmental isolates of S. aureus obtained from institution 1 was established by the UPGMA based on RAPD markers. At both of the institutions, the genera most frequently isolated were Staphylococcus and Bacillus, and the greater concentration of airborne bacteria was detected during the dry season than the rainy season. The lowest diversity and highest dominance was found in the institution 2. On the other hand, at institution 1, the genus that was resistant to antibiotics was Staphylococcus, whereas at institution 2, no isolate was found to be resistant to antibiotics. Furthermore, no association between the clinical and environmental isolates of S. aureus was found. However, one clone was found in different areas of the institution 1. The presence of airborne pathogenic bacteria in institutions 1 and 2 is important to establish the measures for the prevention and control of nosocomial outbreaks.     

Contribution by symbiotically luminous fishes to the occurrence and bioluminescence of luminous bacteria in seawater

Seawater samples from a variety of locations contained viable luminous bacteria, but luminescence was not detectable although the system used to measure light was sensitive enough to measure light from a single, fully induced luminous bacterial cell. When the symbiotically luminous fishCleidopus gloriamaris was placed in a sterile aquarium, plate counts of water samples showed an increase in luminous colony-forming units. Luminescence also increased, decreasing when the fish was removed. Light measurements of water samples from a sterile aquarium containingPhotoblepharon palpebratus, another symbiotically luminous fish, whose bacterial symbionts have not been cultured, showed a similar pattern of increasing light which rapidly decreased upon removal of the fish. These experiments suggest that symbiotically luminous fishes release brightly luminous bacteria from light organs into their environment and may be a source of planktonic luminous bacteria. Although planktonic luminous bacteria are generally not bright when found in seawater, water samples from environments with populations of symbiotically luminous fish may show detectable levels of light.     

Seasonal and Geographic Distribution of Luminous Bacteria in the Eastern Mediterranean Sea and the Gulf of Elat

Luminous bacteria in the Mediterranean Sea and the Gulf of Aqaba-Elat have different distribution patterns. In the Mediterranean Sea, Beneckea harveyi is present all year round, with different subtypes alternating in summer and winter; Photobacterium fischeri was only present during the winter. In the Gulf of Elat, P. leiognathi is present throughout the water column in similar densities during the entire year. This constancy in distribution is presumably due to the near-constancy in water temperature. In summer, Photobacterium leiognathi is replaced by B. harveyi in coastal surface waters. In the hypersaline Bardawil lagoon, only B. harveyi types are present. P. fischeri, a major component of the Mediterranean Sea winter communities, is absent from the lagoon. Luminous Beneckea strains show a great diversity in properties, e.g. temperature range for growth, sensitivity to infection by phages, sensitivity to attack by Bdellovibrio strains, and differences in tolerance to high-salinity shock. Therefore, subdivision of the taxonomic cluster of B. harveyi into subtypes is indicated. The composition of the luminous bacteria communities may serve as indicators of different marine water bodies. The symbiotic luminous bacteria of the light organ of the common Gulf of Elat fish, Photoblepharon palbebratus steinitzi, is different from any of the types described.     

Regulation of aspartokinase activity in the genus Beneckea and marine,luminous bacteria

Summary The pattern of allosteric regulation of aspartokinase activity was determined in species of Beneckea and in the marine, luminous bacteria. The results indicated that these organisms have at least three isofunctional aspartokinases of which the first is inhibited by l-threonine, the second is inhibited by l-lysine, and the third is unaffected by either l-threonine or l-lysine. The homoserine dehydrogenase activity is clearly separable from the l-lysine-sensitive aspartokinase, that may be associated with one (or both) of the other isofunctional aspartokinases.The species of Beneckea and luminous bacteria studied varied in the relative levels of the three isofunctional aspartokinases. B. parahaemolytica, B. alginolytica, B. pelagia, B. neptuna, B. harveyi, B. nigrapulchrituda, B. natriegens, and Photobacterium fischeri had predominant levels of the l-lysine-sensitive activity; B. campellii, P. phosphoreum, and P. mandapamensis had predominant levels of the l-threonine-sensitive activity; while in B. nereida these two activities were approximately equivalent. Species of Beneckea could be distinguished from P. fischeri on the basis of the sensitivity of their aspartokinase activities to inhibition by l-lysine. In P. fischeri 10.3×10^-5 M l-lysine was required for a 50% inhibition of the l-lysine-sensitive enzyme, while in species of Beneckea 0.5–2.7×10^-5 M l-lysine was required to achieve the same level of inhibition.Non-standard abbreviations DAHP 3-deoxy-d-arabino-heptulosonate 7-phosphate - GC guanine plus cytosine     

Characterization of culturable bacterial populations associating with Pinus sylvestris--Suillus bovinus mycorrhizospheres

Bacterial isolations were carried out on Pinus sylvestris--Suillus bovinus mycorrhizospheres obtained directly from boreal pine forest. When samples were taken during dry weather, the numbers of bacterial colony-forming units were significantly higher in uncolonized short roots and external mycelia than in mycorrhizal roots and soil outside the mycorrhizosphere. In contrast, the colony-forming unit counts were similar in all hypogeous samples after rainy weather. Culturable bacteria were absent from most Suillus bovinus sporocarps. The bacteria isolated from all types of mycorr hizo sphere samples, i.e. short roots, mycorrhizal roots, and external mycelia, consisted primarily of Burkholderia spp., whereas most isolates from soil outside the mycorrhizosphere were identified as Paenibacillus spp. This study shows that mycorrhizal external mycelia can expand the habitat favourable for common rhizosphere bacteria into the soil far from the immediate rhizosphere. Some of these bacteria may help the trees with nitrogen acquisition, since potentially diazotrophic bacteria harbouring nitrogenase reductase (nifH) genes were isolated from mycorrhizal root tips.     

Population dynamics and production of the small copepod Oithona spp. in a subarctic fjord of West Greenland

The small cyclopoid copepod Oithona is widely occurring in polar areas; however, knowledge of its biology and ecology is very limited. Here, we investigate the population dynamics, vertical distribution, and reproductive characteristics of Oithona spp. from late winter to summer, in a subarctic fjord of West Greenland. During winter–early spring, the abundance of Oithona spp. was low (1.8 × 10³ ind. m^?2) and the population was mainly composed of late copepodites and adults, whereas in summer, abundance peaked and younger stages dominated (1.1 × 10⁶ ind. m^?2). In general, all stages of Oithona spp. remained in the upper 100 m, with nauplii exhibiting a shallower distribution. Although no general seasonal migration was found, a deeper distribution of the adult females in winter was observed. The mean clutch size of Oithona spp. varied from 16 to 30 eggs per female, peaking in summer. Egg production rates (EPR) were low in winter–early spring (0.13 ± 0.03 eggs female^?1 day^?1) and reached maximum values in summer (1.6 ± 0.45 eggs female^?1 day^?1). EPR of Oithona spp. showed a significantly positive relationship with both temperature and protozooplankton biomass, and the development of the population seemed to be appreciably affected by temperature. Oithona spp. remained active throughout the study, stressing the key importance of these small copepods in high-latitude ecosystems, especially in periods when larger copepods are not present in the surface layer.     

Planktonic Luminous Bacteria of Vellar Estuary,South India

Distribution of planktonic luminous bacteria in relation to environmental parameters was investigated at two stations located in the Vellar Estuary. Luminous microflora showed a pronounced seasonal cycle with very low counts during monsoon months followed by an increase in post monsoon and peak counts during summer. The population density of these procaryotes was remarkably high ranging from 3.5 to 33.1 colony forming units per ml. They constituted 2.1 to 52.1 % of the total bacterial counts. Salinity appeared to govern the distribution of luminous procaryotes as their counts corresponded well with fluctuations in salinity. Taxonomic affiliation of the isolates revealed predominance of Vibrio harveyi. Vibrio fischeri and Photobacterium leiognathi exhibited sparse distribution.     

Spatial and temporal distribution of Caulobacter spp. in two mesotrophic lakes

The seasonal distribution of Caulobacter spp. has been determined in the water column of two mesotrophic lakes using most probable number (MPN) viable counting techniques from April, 1972 to March, 1973. Concentrations in Lake Washington, a monomictic lake, peaked at 1000–3300 per ml in the epilimnion during the late spring and summer and reached lows in October of less than 2 per ml prior to fall turnover. The reason for the decrease in viable numbers is not known. Caulobacter spp. began to increase gradually in December, during the winter mixing period well before the spring bloom. Concentrations in Hall Lake, a small kettle lake, were somewhat higher than in Lake Washington, ranging from 0.27 per ml in October to over 5000 per ml during May. In contrast to Lake Washington, the highest populations were found in the metalimnion and upper hypolimnion of Hall Lake. Caulobacter spp. were also found in the anaerobic depths of the hypolimnion. However, these did not grow anaerobically, suggesting that they had settled into the hypolimnion and survived in the cold anaerobic zone. The populations of Caulobacter spp. reached maximum numbers at the approximate times and depths at which algal biomass would be expected to be greatest.